Monday, November 13, 2017

Genentech's AVASTIN Patents Asserted Against Amgen

On October 10, 2017, Genentech filed a complaint against Amgen alleging that Amgen's biosimilar version of AVASTIN infringes the following patents.

Nitin Balodi of GreyB services helped with the below summary of the patents.


Patent  First Claim
US6054297A 1. A method for making a humanized antibody comprising non-human, import Complementarity Determining Region (CDR) amino acid residues and human Framework Region (FR) amino acid residues, comprising the steps of:
    * (a) obtaining the amino acid sequences of an import variable domain and of a VH subgroup III consensus human variable domain;
    * (b) identifying CDR amino acid sequences in the import and the human variable domain sequences;
    * (c) substituting import CDRs for the corresponding human CDRs;
    * (d) aligning the amino acid sequences of a FR of the import antibody and the corresponding FR of the consensus variable domain;
    * (e) identifying import antibody FR residues in the aligned FR sequences that are non-homologous to the corresponding consensus variable domain residues;
    * (f) determining if the non-homologous import amino acid residue is expected to have at least one of the following effects:
    * (1) non-covalently binds antigen directly;
    * (2) interacts with a CDR; or
    * (3) participates in the VL -VH interface;
    * (g) for any non-homologous import antibody amino acid residue which is expected to have at least one of these effects, substituting that residue for the corresponding amino acid residue in the consensus variable domain FR sequence; and
    * (h) preparing a humanized antibody which binds antigen, wherein the humanized antibody comprises an amino acid sequence determined according to the above steps.
US6121428A 1. A method for recovering a polypeptide comprising:
    * (a) exposing a composition comprising a polypeptide to a reagent which binds to, or modifies, the polypeptide, wherein the reagent is immobilized on a solid phase; and then
    * (b) passing an effluent comprising the polypeptide eluted from or modified by the immobilized reagent, and any reagent leached from the solid phase, through a filter beating a charge which is opposite to the charge of the reagent in and at the pH of, the composition, so as to remove leached reagent from the effluent.
US6242177B1 1. A method of optimizing secretion of a heterologous polypeptide of interest in a cell comprising comparing the levels of expression of the polypeptide under control of a set of nucleic acid variants of a translation initiation region, wherein the set of variants represents a range of translational strengths, and determining the optimal translational strength for production of mature polypeptide, wherein the optimal translational strength is less than the translational strength of the wild-type translation initiation region.
US6331415B1 1. A process for producing an immunoglobulin molecule or an immunologically functional immunoglobulin fragment comprising at least the variable domains of the immunoglobulin heavy and light chains, in a single host cell, comprising the steps of:
(i) transforming said single host cell with a first DNA sequence encoding at least the variable domain of the immunoglobulin heavy chain and a second DNA sequence encoding at least the variable domain of the immunoglobulin light chain, and
(ii) independently expressing said first DNA sequence and said second DNA sequence so that said immunoglobulin heavy and light chains are produced as separate molecules in said transformed single host cell.
US6407213B1 1. A humanized antibody variable domain comprising non-human Complementarity Determining Region (CDR) amino acid residues which bind an antigen incorporated into a human antibody variable domain, and further comprising a Framework Region (FR) amino acid substitution at a site selected from the group consisting of: 4L, 38L, 43L, 44L, 58L, 62L, 65L, 66L, 67L, 68L, 69L, 73L, 85L, 98L, 2H, 4H, 36H, 39H, 43H, 45H, 69H, 70H, 74H, and 92H, utilizing the numbering system set forth in Kabat.
US6417335B1 1. A method for purifying an antibody from a composition comprising the antibody and a contaminant, which method comprises:
(a) loading the composition onto a cation exchange resin, wherein the amount of antibody loaded onto the cation exchange resin is from about 20 mg to about 35 mg of the antibody per mL of cation exchange resin; and
(b) eluting the antibody from the cation exchange resin.
US6586206B1 1. A method of making recombinant proteins using one or more apoptosis inhibitors, comprising the steps of:
(a) providing a vector comprising a gene encoding caspase-9 dominant negative protein,
(b) providing a vector comprising a gene encoding a protein of interest,
(c) providing a Chinese hamster ovary (CHO) host cell,
(d) transforming or transfecting the host cell with the vector of steps (a) and (b),
(e) providing cell culture media,
(f) culturing the transformed or transfected host cell in the cell culture media under conditions sufficient for expression of the protein of interest and the caspase-9 dominant negative protein, and optionally
(g) recovering or purifying the protein of interest from the host cell and/or the cell culture media.
US6620918B2 1. A method for purifying polypeptide monomers from a mixture consisting essentially of said polypeptide monomers, and dimers or multimers of said polypeptide monomers or both dimers and multimers of said polypeptide monomers, wherein the method consists essentially of applying the mixture to a cation-exchange or anion-exchange chromatography resin in a buffer, wherein if the resin is cation-exchange, the pH of the buffer is about 4-7, and wherein if the resin is anion-exchange, the pH of the buffer is about 6-9, and eluting the mixture at a gradient of about 0-1 M of an elution salt, wherein the monomer is purified from the dimers or multimers or both present in the mixture, and wherein the purified monomer has a purity of greater than 99.5% and the monomer yield is greater than 90%.
US6870034B2 1. A method for purifying a protein, which comprises a C H2/C H3 region, from a contaminated solution thereof by Protein A chromatography comprising:
(a) adsorbing the protein from said contaminated solution to Protein A immobilized on a solid phase;
(b) removing contaminants by washing the solid phase with a composition comprising detergent and salt at about pH 4.5 to about 5.5; and
(c) recovering the protein from the solid phase with an elution buffer having a pH in the range from about 2 to about 5.
US6884879B1 1. Isolated nucleic acid encoding a humanized variant of a parent anti-VEGF antibody which parent antibody comprises non-human variable domains, wherein said humanized variant binds human VEGF and comprises the following heavy chain Complementary Determining Region (CDR) amino acid sequences: SEQ ID NO:128 as CDRH1, SEQ ID NO:2 as CDRH2 and SEQ ID NO:129 as CDRH3.
US7060269B1 1. A method for inhibiting VEGF-induced angiogenesis in a subject, comprising administering to said subject an effective amount of a humanized anti-VEGF antibody which binds human VEGF with a Kd  value of no more than about 1×10-8M, said humanized anti-VEGF antibody comprising a heavy chain variable domain sequence of SEQ ID NO:116 and a light chain variable domain sequence of SEQ ID NO:115.
US7169901B2 1. A humanized anti-vascular endothelial growth factor (VEGF) antibody or an antigen binding fragment thereof which binds human VEGF with a Kd value of no more than about 1×10-8M, said humanized anti-VEGF antibody having a heavy a chain variable domain comprising the following heavy chain complementarity determining region (CDR) amino acid sequences: CDRH1 (GYX1FTX2YGMN), wherein X1 is T or D and X2 is N or H; SEQ ID NO:130), CDRH2 (WINTYTGEPTYAADFKR; SEQ ID NO:2) and CDRH3 (YPX1YYGX2SHWYFDV, wherein X1 is Y or H and X2 is S or T; SEQ ID NO:131).
US7375193B2 1. An aqueous formulation useful for inhibiting VEGF-induced angiogenesis in a subject, comprising as the active compound a humanized anti-VEGF antibody in a buffer, wherein the anti-VEGF antibody comprises a heavy chain variable domain comprising the following heavy chain complementarity determining region (CDR) amino acid sequences: CDRH1 (GYTFTNYGMN; SEQ ID NO: 1), CDRH2 (WINTYTGEPTYAADFKR; SEQ ID NO: 2) and CDRH3 (YPHYYGSSHWYFDV; SEQ ID NO: 3) and a light chain variable domain comprising the following light chain CDR amino acid sequences: CDRL1 (SASQDISNYLN; SEQ ID NO: 4), CDRL2 (FTSSLHS; SEQ ID NO: 5) and CDRL3 (QQYSTVPWT; SEQ ID NO: 6).
US7622115B2 1. A method for treating cancer in a patient comprising administering an effective amount of bevacizumab and assessing the patient for gastrointestinal perforation during treatment with bevacizumab.
US7807799B2 1. A method of purifying a protein which comprises a CH2/CH3 region, comprising subjecting a composition comprising said protein to protein A affinity chromatography at a temperature in the range from about 10 ° C. to about 18 ° C.
US7923221B1 1. A method for making an antibody heavy chain or fragment thereof and an antibody light chain or fragment thereof each having specificity for a desired antigen, wherein the heavy chain or fragment thereof comprises a human constant region sequence and a variable region comprising non human mammalian variable region sequences, the method comprising culturing a recombinant host cell comprising DNA encoding the heavy chain or fragment thereof and the light chain or fragment thereof and recovering the heavy chain or fragment thereof and light chain or fragment thereof from the host cell culture.
US8044017B2 1. A method for purifying a polypeptide from a composition comprising the polypeptide and contaminants, which method comprises the sequential steps of:
(a) loading the composition onto an ion exchange resin with an equilibration buffer having a first salt concentration;
(b) washing the ion exchange resin with a wash buffer until a predetermined protein concentration is measured in the flowthrough, wherein the salt concentration of the wash buffer increases from an initial, second salt concentration that is greater than the salt concentration of the equilibration buffer, to a final, third salt concentration;
(c) passing a fixed volume of wash buffer at the final, third salt concentration over the cation exchange resin; and
(d) eluting the polypeptide from the ion exchange resin with elution buffer that has a salt concentration that is greater than the final salt concentration of the wash buffer.
US8460895B2 1. A method for the recombinant production of a polypeptide in a eukaryotic host cell modified in the citrate cycle to express a cytosolic pyruvate carboxylase, the method comprising the following steps:
(a) cultivating the eukaryotic host cell in a suitable medium under conditions which allow the expression of the polypeptide, wherein the content of dissolved CO 2 in the medium is maintained at a constant value in the range of 10% to 20% of the saturated solution of CO2 under a given set of conditions, and wherein a base is added to adjust the pH of the medium; and
(b) recovering the polypeptide from the cell or from the medium.
US8512983B2 1. A process for producing a polypeptide in a mammalian host cell expressing said polypeptide, comprising culturing the mammalian host cell in a production phase of the culture in a glutamine-free production culture medium containing asparagine, wherein the asparagine is added at a concentration in the range of 7.5 mM to 15 mM.
US8574869B2 1. A method for the prevention of the reduction of a disulfide bond in an antibody expressed in a recombinant host cell, comprising, following fermentation, sparging the pre-harvest or harvested culture fluid of said recombinant host cell with air, wherein the amount of dissolved oxygen (dO2) in the pre-harvest or harvested culture fluid is at least 10%.
US8633302B2 1. A method for concentrating an immunoglobulin solution by tangential flow filtration, characterized in that the transmembrane pressure and the cross-flow are variable, and changed during the filtration process according to the concentration of the immunoglobulin to be concentrated, wherein
i) a transmembrane pressure of from 1.4 bar to 1.6 bar and a cross-flow of from 75 ml/min. to 90 ml/min. is applied in a concentration range up to 30 mg immunoglobulin per ml of solution to be concentrated,
ii) a transmembrane pressure of from 0.8 bar to 0.9 bar and a cross-flow of from 140 ml/min. to 160 ml/min. is applied in a concentration range of from 15 mg/ml up to 55 mg/ml, and
iii) a transmembrane pressure of from 0.8 bar to 0.9 bar and a cross-flow of from 120 ml/min. to 140 ml/min is applied in a concentration range of more than 45 mg/ml up to about 130 mg/ml.
US8710196B2 1. A method for purifying an antibody from a composition comprising the antibody and a contaminant, which method comprises the following steps performed sequentially:
(a) binding the antibody to a cation exchange material with an equilibration buffer at a first conductivity;
(b) washing the cation exchange material with a wash buffer, wherein the conductivity of the wash buffer increases from a second conductivity that is higher than the first conductivity to a third conductivity during the washing;
(c) passing a fixed volume of wash buffer at the third conductivity over the cation exchange material; and
(d) eluting the antibody from the cation exchange material with an elution buffer at a fourth conductivity that is higher than the third conductivity.
US9441035B2 1. A method of producing bevacizumab, or a fragment thereof, comprising the step of culturing a Chinese hamster ovary (CHO) cell comprising a nucleic acid encoding bevacizumab or fragment thereof in a cell culture medium, wherein the cell culture medium comprises copper, insulin, and cystine, wherein the cystine is at a concentration of from 1.25 mM to 2.5 mM, and wherein the cell produces bevacizumab, or a fragment thereof.
US9487809B2 1. A method for reducing lactate production in cultured cells, the method comprising culturing cells comprising a first heterologous nucleic acid sequence encoding a small interfering RNA (siRNA) specific for a lactate dehydrogenase (LDH) and a second heterologous nucleic acid sequence encoding an siRNA specific for a pyruvate dehydrogenase kinase (PDHK), wherein the first heterologous nucleic acid sequence is operably linked to a first promoter, and wherein the second heterologous nucleic acid sequence is operably linked to a second promoter, wherein the cultured cells have a polypeptide productivity of at least about 68% higher than cultured cells without the heterologous nucleic acid sequence comprising siRNA specific for PDHK and the siRNA specific for LDH.

Friday, November 10, 2017

HUMIRA Patents Asserted Against Boehringer

On August 2, 2017, AbbVie filed a patent infringement suit (1:17-cv-01065 (Del)) against Boehringer, alleging that Boehringer's biosimilar version of HUMIRA infringes AbbVie's  patents.  Abbvie provides an extensive list of over seventy patents in its complaint, but only asserted eight of them: "Because of Boehringer’s actions, AbbVie is limited to asserting the following eight patents in the present lawsuit: U.S. Patent No. 8,926,975; U.S. Patent No. 9,018,361; U.S. Patent No. 9,090,867; U.S. Patent No. 9,096,666; U.S. Patent No. 9,255,143; U.S. Patent No. 9,266,949; U.S. Patent No. 9,272,041; and U.S. Patent No. 9,546,212."

The following list of patents was put together by Nitin Balodi of GreyB services (nitin.balodi@greyb.com).


Patent NumberFirst Claim
US82318761. A host cell protein (HCP)-reduced antibody preparation produced by a method comprising:
applying a mixture comprising an antibody and at least one HCP to a cation exchange resin in an equilibration buffer, wherein greater than 30 grams of antibody per liter of cation exchange resin are applied;
washing HCP from the cation exchange resin with a plurality of wash steps comprising a first wash and a second wash, wherein there is an increase in conductivity from the first wash to the second wash;
eluting the antibody from the cation exchange resin with an elution buffer to form a first eluate;
applying the first eluate to an anion exchange resin, wherein prior to applying the first eluate to the anion exchange resin, pH and conductivity of the first eluate are adjusted to be substantially similar to pH and conductivity of the anion exchange resin; and
obtaining a first flowthrough comprising the antibody, such that the HCP-reduced antibody preparation is obtained,
wherein the antibody is an isolated human anti-TNFα antibody that dissociates from human TNFα with a K d of 1×10−8 M or less and a Koff rate constant of 1×10−3 s−1 or less, both determined by surface plasmon resonance, and neutralizes human TNFα cytotoxicity in a standard in vitro L929 assay with an IC50 of 1×10−7 M or less, and
wherein the HCP-reduced antibody preparation comprises no greater than about 70 ng of HCP per mg of antibody as measured by a HCP ELISA and a cathepsin L activity of no greater than about 3.0 RFU/s/mg antibody.
US86639451. A method of producing an anti-TNFα antibody in a mammalian cell culture, said method comprising:
a) culturing Chinese Hamster Ovary (CHO) cells comprising a nucleic acid encoding the antibody, or a fragment thereof, in a cell culture growth medium to form the mammalian cell culture; and
b) culturing the CHO cells in a cell culture production medium, wherein glucose is added to the mammalian cell culture to maintain a glucose concentration of at least 2 g/L,
such that the anti-TNFα antibody is produced,
wherein the anti-TNFα antibody comprises a light chain variable region (LCVR) having a CDR3 domain comprising the amino acid sequence of SEQ ID NO:3, a CDR2 domain comprising the amino acid sequence of SEQ ID NO:5, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO: 7, and comprises a heavy chain variable region (HCVR) having a CDR3 domain comprising the amino acid sequence of SEQ ID NO:4, a CDR2 domain comprising the amino acid sequence of SEQ ID NO: 6, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO:8.
US87156641. A method for inhibiting radiographic disease progression in a human subject having erosive polyarthritis associated with a disorder in which TNFα activity is detrimental, comprising administering to the subject having erosive polyarthritis an isolated human anti-TNFα antibody, or antigen-binding portion thereof, such that radiographic disease progression is inhibited in the subject, wherein a Total Sharp Score (TSS) of the subject is either maintained or decreased following administration of the human anti-TNFα antibody, or antigen-binding portion thereof, wherein the human anti-TNFα antibody, or an antigen-binding portion thereof, dissociates from human TNFα with a Kd of 1×10−8 M or less and a Koff rate constant of 1×10−3 s−1 or less, both determined by surface plasmon resonance, and neutralizes human TNFα cytotoxicity in a standard in vitro L929 assay with an IC50 of 1×10−7 M or less.
US88021001. A stable liquid aqueous pharmaceutical formulation comprising
(a) a human IgG1 anti-human Tumor Necrosis Factor alpha (TNFα) antibody, or an antigen-binding portion thereof, at a concentration of 45 to 150 mg/ml,
(b) a polyol,
(c) a polysorbate at a concentration of 0.1 to 10 mg/ml, and
(d) a buffer system having a pH of 4.5 to 7.0,
wherein the antibody comprises the light chain variable region and the heavy chain variable region of D2E7.
US88021011. A stable liquid aqueous pharmaceutical formulation comprising
(a) a human IgG1 anti-human Tumor Necrosis Factor alpha (TNFα) antibody, or an antigen-binding portion thereof, at a concentration of 45 to 105 mg/ml,
(b) a polyol,
(c) a polysorbate at a concentration of 0.1 to 10 mg/ml, and
(d) a buffer system comprising acetate and having a pH of 4.5 to 7.0,
wherein the antibody comprises the light chain variable region and the heavy chain variable region of D2E7.
US8808700B11. A method for treating erosive polyarthritis, comprising administering a human anti-TNFα antibody to a human subject having erosive polyarthritis associated with psoriatic arthritis, ankylosing spondylitis or juvenile rheumatoid arthritis, wherein a modified Total Sharp Score (mTSS) of the subject is maintained or decreased following said treating as compared to baseline prior to said treating, and wherein the human anti-TNFα antibody comprises (1) a light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO: 1 and (2) a heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO: 2.
US88831561. A method of treating a disorder in which TNFα activity is detrimental in a subject, the method comprising administering a composition comprising a therapeutically effective amount of adalimumab to the subject such that the disorder is treated,
wherein the adalimumab is produced in a Chinese Hamster Ovary (CHO) cell expression system;
wherein the disorder is selected from the group consisting of rheumatoid arthritis, Crohn's disease, ulcerative colitis, ankylosing spondylitis, psoriatic arthritis, psoriasis, and juvenile rheumatoid arthritis; and
wherein the composition is characterized in that when the composition is assayed in a cathepsin L kinetic assay, a level of cathepsin L activity from 0.4 to less than 1.84 RFU/s/mg of adalimumab is observed, wherein the cathepsin L kinetic assay comprises:
i) diluting the composition in a polystyrene container in a solution containing 25 mM NaOAc, 5 mM DTT and 1 mM EDTA at pH 5.5,
ii) adding dextran sulfate to a concentration of 0.035 μg/mL and incubating at 37° C. for six hours,
iii) adding Z-leucine-arginine covalently bound at its C-terminus to a fluorescent 7-amino-4-methyl coumarin (Z-leucine-arginine-AMC), wherein the diluting, adding, and incubating steps are sufficient to permit the measurement of cathepsin L hydrolysis of the Z-leucine-arginine-AMC within a linear range, and
iv) measuring Z-leucine-arginine-AMC hydrolysis in the linear range in RFU/s/mg of adalimumab.
US88891351. A method for treating rheumatoid arthritis in a human subject, comprising administering subcutaneously to a human subject having rheumatoid arthritis a total body dose of 40 mg of a human anti-TNFα antibody once every 13 -15 days for a time period sufficient to treat the rheumatoid arthritis, wherein the anti-TNFα antibody comprises an IgG1 heavy chain constant region; a variable light (“VL”) chain region comprising a CDR1 having the amino acid sequence of SEQ ID NO:7, a CDR2 having the amino acid sequence of SEQ ID NO:5, and a CDR3 having the amino acid sequence of SEQ ID NO:3; and a variable heavy (“VH”) chain region comprising a CDR1 having the amino acid sequence of SEQ ID NO:8, a CDR2 having the amino acid sequence of SEQ ID NO:6 and a CDR3 having the amino acid sequence of SEQ ID NO:4.
US88891361. A multiple-variable dose method for inducing clinical remission of Crohn's disease in a subject in need thereof, comprising subcutaneously administering to the subject:
a first dose of 160 mg of a recombinant human anti-TNFα antibody administered as a set of four injections of 40 mg of the antibody administered to the subject within a day; and
a second dose of 80 mg of the antibody administered as a set of two injections of 40 mg of the antibody administered to the subject within a day, wherein the second dose is administered two weeks following administration of the first dose;
wherein the antibody comprises:
a heavy chain comprising a CDR1 comprising the amino acid sequence of SEQ ID NO:8; a CDR2 comprising the amino acid sequence of SEQ ID NO:6; and a CDR3 comprising the amino acid sequence of SEQ ID NO:4; and
a light chain comprising a CDR1 comprising the amino acid sequence of SEQ ID NO:7; a CDR2 comprising the amino acid sequence of SEQ ID NO:5; and a CDR3 comprising the amino acid sequence of SEQ ID NO:3.
US8895009B21. A liquid composition comprising adalimumab, wherein the adalimumab is expressed in a Chinese Hamster Ovary (CHO) cell expression system; and wherein the composition is characterized in that when the composition is assayed in a cathepsin L kinetic assay, a level of cathepsin L activity from 0.4 to less than 1.84 RFU/s/mg of adalimumab is observed, wherein the cathepsin L kinetic assay comprises:
i) diluting the composition in a polystyrene container in a solution containing 25 mM NaOAc, 5 mM DTT and 1 mM EDTA at pH 5.5,
ii) adding dextran sulfate to a concentration of 0.035 μg/mL and incubating at 37° C. for six hours,
iii) adding Z-leucine-arginine covalently bound at its C-terminus to a fluorescent 7-amino-4-methyl coumarin (Z-leucine-arginine-AMC), wherein the diluting, adding, and incubating steps are sufficient to permit the measurement of cathepsin L hydrolysis of the Z-leucine-arginine-AMC within a linear range, and
iv) measuring Z-leucine-arginine-AMC hydrolysis in the linear range in RFU/s/mg of adalimumab.
US89063721. A method of treating a disorder in which TNFα activity is detrimental in a subject, the method comprising administering a composition comprising a therapeutically effective amount of adalimumab to the subject such that the disorder is treated,
wherein the adalimumab is produced in a Chinese Hamster Ovary (CHO) cell expression system;
wherein the disorder is selected from the group consisting of rheumatoid arthritis, Crohn's disease, ulcerative colitis, ankylosing spondylitis, psoriatic arthritis, psoriasis, and juvenile rheumatoid arthritis; and
wherein the composition is characterized in that when the composition is assayed in a cathepsin L kinetic assay, a level of cathepsin L activity from 0.6 to less than 1.84 RFU/s/mg of adalimumab is observed, wherein the cathepsin L kinetic assay comprises:
i) diluting the composition in a polystyrene container in a solution containing 25 mM NaOAc, 5 mM DTT and 1 mM EDTA at pH 5.5,
ii) adding dextran sulfate to a concentration of 0.035 μg/mL and incubating at 37° C. for six hours,
iii) adding Z-leucine-arginine covalently bound at its C-terminus to a fluorescent 7-amino-4-methyl coumarin (Z-leucine-arginine-AMC), wherein the diluting, adding, and incubating steps are sufficient to permit the measurement of cathepsin L hydrolysis of the Z-leucine-arginine-AMC within a linear range, and
iv) measuring Z-leucine-arginine-AMC hydrolysis in the linear range in RFU/s/mg of adalimumab.
US89063731. A method of treatment of psoriasis in a patient with moderate to severe active psoriatic arthritis (PsA), comprising subcutaneously administering to said patient 40 mg adalimumab every other week, wherein said patient has ≧3 swollen and ≧3 tender joints prior to the treatment and has failed NSAID therapy, and wherein said patient achieves a 90% reduction of the Psoriasis Area and Severity Index Score (PASI 90) at week 24 of the treatment.
US89066461. A fed-batch method of making a human anti-TNFα antibody comprising a light chain variable region (LCVR) comprising the sequence of SEQ ID NO:1 and a heavy chain variable region (HCVR) comprising the sequence of SEQ ID NO:2, said method comprising culturing Chinese Hamster Ovary (CHO) cells comprising a nucleic acid encoding said LCVR and HCVR of said anti-TNFα antibody in a cell culture production medium in large-scale,
wherein glucose concentration in said cell production medium is monitored, the glucose concentration in said medium decreases to below 2 g/L, and glucose is added to said medium when the glucose concentration in said medium decreases to below 2 g/L, or the glucose concentration in said medium is monitored and glucose is added to said medium to maintain the glucose concentration in said medium at a concentration of at least 2 g/L but no greater than 7 g/L, such that said anti-TNFα antibody is produced, and wherein said produced antibody is further affinity purified using a Protein A resin.
US89117371. A method for treating Crohn's disease in a human subject, comprising administering subcutaneously to a human subject having Crohn's disease a total body dose of 40 mg of a human anti-TNFα antibody once every 13-15 days for a time period sufficient to treat Crohn's disease, wherein the anti-TNFα antibody comprises an IgG1 heavy chain constant region; a variable light (“VL”) chain region comprising a CDR1 having the amino acid sequence of SEQ ID NO:7, a CDR2 having the amino acid sequence of SEQ ID NO:5, and a CDR3 having the amino acid sequence of SEQ ID NO:3; and a variable heavy (“VH”) chain region comprising a CDR1 having the amino acid sequence of SEQ ID NO: 8, a CDR2 having the amino acid sequence of SEQ ID NO:6 and a CDR3 having the amino acid sequence of SEQ ID NO:4.
US89119641. A fed-batch production method of making a human anti-TNFα antibody which comprises (1) a light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO:1 and (2) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:2, said method comprising culturing Chinese Hamster Ovary (CHO) cells comprising a nucleic acid encoding said anti-TNFα antibody in a cell culture production medium in large-scale, wherein the glucose concentration in said medium is monitored, the glucose concentration in said medium decreases to below 2 g/L, and glucose is added to said medium when the glucose concentration in said medium decreases to below 2 g/L, or the glucose concentration in said medium is monitored and glucose is added to said medium to maintain the glucose concentration in said medium at a concentration of at least 2 g/L but no greater than 7 g/L, such that said anti-TNFα antibody is produced at a titer of at least 2 g/L in said cell culture production medium.
US89161531. A pharmaceutical composition comprising adalimumab and a pharmaceutically acceptable carrier, wherein the adalimumab is produced in a Chinese Hamster Ovary (CHO) cell expression system; and wherein the composition is characterized in that when the composition is assayed in a cathepsin L kinetic assay, a level of cathepsin L activity from 0.4 to less than 1.84 RFU/s/mg of adalimumab is observed, wherein the cathepsin L kinetic assay comprises:
i) diluting the composition in a polystyrene container in a solution containing 25 mM NaOAc, 5 mM DTT and 1 mM EDTA at pH 5.5,
ii) adding dextran sulfate to a concentration of 0.035 μg/mL and incubating at 37° C. for six hours,
iii) adding Z-leucine-arginine covalently bound at its C-terminus to a fluorescent 7-amino-4-methyl coumarin (Z-leucine-arginine-AMC), wherein the diluting, adding, and incubating steps are sufficient to permit the measurement of cathepsin L hydrolysis of the Z-leucine-arginine-AMC within a linear range, and
iv) measuring Z-leucine-arginine-AMC hydrolysis in the linear range in RFU/s/mg of adalimumab.
US89161571. A stable liquid aqueous pharmaceutical formulation comprising
(a) a human IgG1 anti-human Tumor Necrosis Factor alpha (TNFα) antibody, or an antigen-binding portion thereof, at a concentration of 20 to 150 mg/ml,
(b) a tonicity agent,
(c) a surfactant, and
(d) a buffer system having a pH of 4.0 to 8.0,
wherein the antibody comprises the light chain variable region and the heavy chain variable region of D2E7.
US89161581. A stable liquid aqueous pharmaceutical formulation comprising
(a) a human IgG1 anti-human Tumor Necrosis Factor alpha (TNFα) antibody, or an antigen-binding portion thereof, at a concentration of 20 to 150 mg/ml,
(b) a polyol,
(c) a surfactant, and
(d) a buffer system having a pH of 4 to 8,
wherein the antibody comprises the light chain variable region and the heavy chain variable region of D2E7.
US89269751. A method of treating active ankylosing spondylitis (AS) in a subject having total spinal ankylosis comprising selecting a subject having active AS and total spinal ankylosis, and subcutaneously administering 40 mg of an isolated human anti-TNFα antibody to the subject once every other week, wherein the human anti-TNFα antibody comprises a light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO: 1, and comprises a heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO: 2.
US89463951. A method for producing a preparation comprising a protein of interest and having a reduced level of at least one impurity, said method comprising:
(a) contacting a sample comprising the protein of interest and at least one impurity, to a hydrophobic interaction chromatography (HIC) media, in the presence of a load buffer such that (i) a portion of the protein of interest binds to the HIC media at a Kp of greater than 20 and (ii) a substantial portion of the at least one impurity binds to the HIC media;
(b) collecting a flow through fraction comprising the protein of interest unbound to the HIC media;
(c) washing the HIC media with a wash buffer such that a substantial portion of the protein of interest bound to the HIC media is released from the media, wherein the salt concentration and/or the pH of the wash buffer are within 20% of the salt concentration and/or pH of the load buffer; and
(d) collecting a wash fraction comprising the protein of interest released from the HIC media,
wherein each of the flow through and wash fractions comprise the protein of interest and have a reduced level of the at least one impurity.
US89619731. A multiple-variable dose method for inducing clinical remission of Crohn's disease in a subject in need thereof, comprising subcutaneously administering to the subject:
a first dose of 160 mg of a recombinant human anti-TNFα antibody administered to the subject within a day; and
a second dose of 80 mg of the antibody administered to the subject within a day, wherein the second dose is administered two weeks following administration of the first dose;
wherein the antibody comprises:
a heavy chain comprising a CDR1 comprising the amino acid sequence of SEQ ID NO:8; a CDR2 comprising the amino acid sequence of SEQ ID NO:6; and a CDR3 comprising the amino acid sequence of SEQ ID NO:4; and
a light chain comprising a CDR1 comprising the amino acid sequence of SEQ ID NO:7; a CDR2 comprising the amino acid sequence of SEQ ID NO:5; and a CDR3 comprising the amino acid sequence of SEQ ID NO:3.
US896197421. A multiple-variable dose method for treating ulcerative colitis in a subject in need thereof, comprising subcutaneously administering to the subject:
a first dose of 160 mg of a recombinant human anti-TNFα antibody administered to the subject within a day; and
a second dose of 80 mg of the antibody administered to the subject within a day, wherein the second dose is administered two weeks following administration of the first dose;
wherein the antibody comprises:
a heavy chain comprising a CDR1 comprising the amino acid sequence of SEQ ID NO:8; a CDR2 comprising the amino acid sequence of SEQ ID NO:6; and a CDR3 comprising the amino acid sequence of SEQ ID NO:4; and
a light chain comprising a CDR1 comprising the amino acid sequence of SEQ ID NO:7; a CDR2 comprising the amino acid sequence of SEQ ID NO:5; and a CDR3 comprising the amino acid sequence of SEQ ID NO:3.
US89747901. A method for treating ulcerative colitis in a human subject, comprising administering subcutaneously to a human subject having ulcerative colitis a total body dose of 40 mg of a human anti-TNFα antibody once every 13-15 days for a time period sufficient to treat the ulcerative colitis, wherein the anti-TNFα antibody comprises an IgG1 heavy chain constant region; a variable light (“VL”) chain region comprising a CDR1 having the amino acid sequence of SEQ ID NO:7, a CDR2 having the amino acid sequence of SEQ ID NO:5, and a CDR3 having the amino acid sequence of SEQ ID NO:3; and a variable heavy (“VH”) chain region comprising a CDR1 having the amino acid sequence of SEQ ID NO:8, a CDR2 having the amino acid sequence of SEQ ID NO:6and a CDR3 having the amino acid sequence of SEQ ID NO:4.
US89866931. A method for treating moderate to severe chronic plaque psoriasis, comprising subcutaneously administering to an adult patient having moderate to severe chronic plaque psoriasis a first dose of 80 mg of adalimumab, followed by 40 mg of adalimumab every other week starting one week after said first dosing, wherein the patient achieves at least Psoriasis Area and Severity Index (PASI) 90 response at week 12 of the treatment.
US89929261. A method for treating rheumatoid spondylitis in a human subject, comprising administering subcutaneously to a human subject having rheumatoid spondylitis a total body dose of 40 mg of a human anti-TNFα antibody once every 13-15 days for a time period sufficient to treat the rheumatoid spondylitis, wherein the anti-TNFα antibody comprises an IgG1 heavy chain constant region; a variable light (“VL”) chain region comprising a CDR1 having the amino acid sequence of SEQ ID NO:7, a CDR2 having the amino acid sequence of SEQ ID NO:5, and a CDR3 having the amino acid sequence of SEQ ID NO:3; and a variable heavy (“VH”) chain region comprising a CDR1 having the amino acid sequence of SEQ ID NO:8, a CDR2 having the amino acid sequence of SEQ ID NO:6 and a CDR3 having the amino acid sequence of SEQ ID NO:4.
US89993371. A method for treating juvenile idiopathic arthritis (JIA) in a subject comprising administering an isolated human anti-TNFα antibody, or an antigen-binding portion thereof, to the subject, wherein 20 mg of the anti-TNFα antibody, or antigen-binding portion thereof, is administered to the subject if the subject weighs less than 30 kg, or wherein 40 mg of the anti-TNFα antibody, or antigen-binding portion thereof, is administered to the subject if the subject weighs more than or equal to 30 kg.
US90176801. A method of reducing signs and symptoms in a patient with moderately to severely active rheumatoid arthritis, comprising:
administering to said patient, in combination with methotrexate, a human anti-TNFα antibody,
wherein the human anti-TNFα antibody is administered subcutaneously in a total body dose of 40 mg once every 13-15 days, and
wherein the anti-TNFα antibody comprises an IgG1 heavy chain constant region; a variable light (“V L”) chain region comprising a CDR1 having the amino acid sequence of SEQ ID NO:7, a CDR2 having the amino acid sequence of SEQ ID NO:5, and a CDR3 having the amino acid sequence of SEQ ID NO:3; and a variable heavy (“VH”) chain region comprising a CDR1 having the amino acid sequence of SEQ ID NO:8, a CDR2 having the amino acid sequence of SEQ ID NO:6 and a CDR3 having the amino acid sequence of SEQ ID NO:4.
US90183611. A process for purifying adalimumab from a fermentation harvest of a Chinese Hamster Ovary (CHO) cell culture expressing said adalimumab, said process comprising:
a) binding adalimumab from said fermentation harvest to a Protein A resin,
b) eluting the bound adalimumab at an elution pH of 3.6-4, and
c) incubating the eluted adalimumab for 1 to 3 hours.
US90610051. A multiple-variable dose method for treating idiopathic inflammatory bowel disease in a subject in need thereof, comprising subcutaneously administering to the subject:
a first dose of 160 mg of a recombinant human anti-TNFα antibody administered to the subject within a day; and
a second dose of 80 mg of the antibody administered to the subject within a day, wherein the second dose is administered two weeks following administration of the first dose;
wherein the antibody comprises:
a heavy chain comprising a CDR1 comprising the amino acid sequence of SEQ ID NO:8; a CDR2 comprising the amino acid sequence of SEQ ID NO:6; and a CDR3 comprising the amino acid sequence of SEQ ID NO:4; and
a light chain comprising a CDR1 comprising the amino acid sequence of SEQ ID NO:7; a CDR2 comprising the amino acid sequence of SEQ ID NO:5; and a CDR3 comprising the amino acid sequence of SEQ ID NO:3.
US90621061. A method for increasing the galactosylation level of a recombinantly expressed adalimumab, comprising supplementing a media used in the expression of the recombinantly expressed adalimumab with a sufficient amount of a manganese supplement to achieve a manganese concentration in the media of 0.2-100 μM and a sufficient amount of a galactose supplement to achieve a galactose concentration in the media of 1-100 mM, thereby increasing the galactosylation level of the recombinantly expressed adalimumab, wherein the galactosylation level of a recombinantly expressed adalimumab is increased as compared to the galactosylation level of adalimumab recombinantly expressed in media which is not supplemented with said manganese supplement and said galactose supplement.
US90679921. A method of treatment of moderate to severe active psoriatic arthritis in adult patients, wherein each said patient has ≧3 swollen and ≧3 tender joints prior to the treatment and has failed NSAID therapy, comprising subcutaneously administering to each said patient 40 mg of adalimumab every other week, wherein 23% of said patients achieve 70% reduction in American College of Rheumatology (ACR) score at week 24 of the treatment.
US90739871. A method of reducing signs and symptoms in a patient with moderately to severely active rheumatoid arthritis, comprising:
administering to said patient a total body dose of 40 mg of a human anti-TNFα antibody,
wherein the dose is administered subcutaneously from a 40 mg dosage unit form once every 13-15 days, and
wherein the anti-TNFα antibody comprises an IgG1 heavy chain constant region; a variable light (“V L”) chain region comprising a CDR1 having the amino acid sequence of SEQ ID NO:7, a CDR2 having the amino acid sequence of SEQ ID NO:5, and a CDR3 having the amino acid sequence of SEQ ID NO:3; and a variable heavy (“VH”) chain region comprising a CDR1 having the amino acid sequence of SEQ ID NO:8, a CDR2 having the amino acid sequence of SEQ ID NO:6 and a CDR3 having the amino acid sequence of SEQ ID NO:4.
US90739881. A fed batch method for making an anti-TNFα antibody comprising a light chain variable region (LCVR) comprising the sequence of SEQ ID NO:1 and a heavy chain variable region (HCVR) comprising the sequence of SEQ ID NO:2, said method comprising culturing mammalian cells comprising a nucleic acid encoding said anti-TNFα antibody in a cell culture production medium in large scale, wherein the pH of the cell culture production medium is adjusted according to a pH linear ramp comprising beginning at a starting pH and ending at a final pH that is less than the starting pH, such that said anti-TNFα antibody is produced, and wherein said produced anti-TNFα antibody is further affinity purified using a Protein A resin.
US90856181. A low acidic species composition comprising adalimumab, wherein the composition comprises less than 10% total acidic species of adalimumab and wherein less than 75% of the lysine variant species of the composition have zero C-terminal lysines (Lys 0), and wherein the acidic species of adalimumab are quantified based on the relative area percent of peaks that elute earlier than the main peak in a WCX-10 HPLC chromatogram of adalimumab wherein the WCX-10 HPLC chromatogram is generated using a first mobile phase of 10mM Sodium Phosphate dibasic (pH 7.5) and a second mobile phase of 10mM Sodium Phosphate dibasic, 500 mM Sodium Chloride (pH 5.5) and wherein the WCX-10 HPLC chromatogram is generated using detection at 280 nm.
US90856201. A method of administering adalimumab for treatment of psoriatic arthritis, comprising filling adalimumab into vessels and subcutaneously administering 40 mg of said adalimumab to a patient having psoriatic arthritis every other week.
US90906881. A process for producing an antibody comprising the heavy and light chain variable domains of adalimumab, the process comprising culturing a mammalian cell which produces said antibody in a cell culture media that comprises a concentration of manganese of 0.2-100 μM and a concentration of galactose of 1-100 mM, wherein said concentration of manganese and galactose is sufficient to increase the galactosylation level of said antibody as compared to the galactosylation level of an antibody comprising the heavy and light chain variable domains of adalimumab produced in a cell culture media that does not comprise said concentration of manganese and galactose, thereby producing said antibody.
US90906891. A method of administering adalimumab for treatment of moderate to severe chronic plaque psoriasis, comprising filling adalimumab into vessels and subcutaneously administering 40 mg of said adalimumab to a patient having moderate to severe chronic plaque psoriasis every other week.
US90908671. A fed-batch method for making an anti-TNFα antibody comprising a light chain variable region (LCVR) comprising the sequence of SEQ ID NO:1 and a heavy chain variable region (HCVR) comprising the sequence of SEQ ID NO:2, said method comprising culturing mammalian cells comprising a nucleic acid encoding said anti-TNFα antibody in a cell culture production medium in large scale, wherein the pH of the cell culture production medium is adjusted such that the culturing begins at a starting pH and ends at a final pH that is less than the starting pH, such that said anti-TNFαantibody is produced at a titer of at least 2 g/L in said cell culture production medium.
US90966661. A liquid composition comprising adalimumab, wherein the adalimumab is expressed in a Chinese Hamster Ovary (CHO) cell expression system; and the composition is characterized in that when the composition is assayed in a cathepsin L kinetic assay, a level of cathepsin L activity less than 1.84 RFU/s/mg of adalimumab is observed, wherein the cathepsin L kinetic assay comprises:
i) diluting the composition in a polystyrene container in a solution containing 25 mM NaOAc, 5 mM DTT and 1 mM EDTA at pH 5.5,
ii) adding dextran sulfate to a concentration of 0.035 μg/mL and incubating at 37° C. for six hours,
iii) adding Z-leucine-arginine covalently bound at its C-terminus to a fluorescent 7-amino-4-methyl coumarin (Z-leucine-arginine-AMC), wherein the diluting, adding, and incubating steps are sufficient to permit the measurement of cathepsin L hydrolysis of the Z-leucine-arginine-AMC within a linear range, and
iv) measuring Z-leucine-arginine-AMC hydrolysis in the linear range in RFU/s/mg of adalimumab.
US91027231. A method for preparing an adalimumab formulation, comprising the step of mixing a liquid composition comprising adalimumab with a pharmaceutically acceptable carrier, wherein the adalimumab is expressed in a Chinese Hamster Ovary (CHO) cell expression system, and the composition is characterized in that when the composition is assayed in a cathepsin L kinetic assay, a level of cathepsin L activity of less than 1.84 RFU/s/mg of adalimumab is observed, wherein the cathepsin L kinetic assay comprises:
i) diluting the composition in a polystyrene container in a solution containing 25 mM NaOAc, 5 mM DTT and 1 mM EDTA at pH 5.5,
ii) adding dextran sulfate to a concentration of 0.035 μg/mL and incubating at 37° C. for six hours,
iii) adding Z-leucine-arginine covalently bound at its C-terminus to a fluorescent 7-amino-4-methyl coumarin (Z-leucine-arginine-AMC), wherein the diluting, adding, and incubating steps are sufficient to permit the measurement of cathepsin L hydrolysis of the Z-leucine-arginine-AMC within a linear range, and
iv) measuring Z-leucine-arginine-AMC hydrolysis in the linear range in RFU/s/mg of adalimumab.
US91141661. A stable liquid aqueous pharmaceutical formulation comprising: a human anti-human Tumor Necrosis Factor alpha (TNFα) IgG1 antibody at a concentration of 50 mg/ml, wherein the antibody comprises the light chain variable region and the heavy chain variable region of D2E7, and a buffer system;
wherein the formulation is isotonic, suitable for single-use subcutaneous injection, and has a pH of 4.0 to 8.0.
US91506451. A method for producing a composition comprising adalimumab, the method comprising culturing a mammalian cell producing adalimumab in cell culture media comprising 2 g/L to 11 g/L of each of one or more basic amino acids selected from the group consisting of arginine, lysine, ornithine and histidine, and combinations thereof, to produce a composition comprising adalimumab, wherein the composition comprises less than 20% total acidic species of adalimumab, wherein the acidic species of adalimumab do not include process-related impurities selected from the group consisting of host cells and lysed host cells and wherein the acidic species of adalimumab correspond to the peaks that elute earlier than the main peak in a WCX-10 HPLC chromatogram of adalimumab, and wherein the WCX-10 HPLC chromatogram is generated using a first mobile phase of 10 mM Sodium Phosphate dibasic (pH 7.5) and a second mobile phase of 10 mM Sodium Phosphate dibasic, 500 mM Sodium Chloride (pH 5.5), and wherein the WCX-10 HPLC chromatogram is generated using detection at 280 nm.
US91875591. A multiple-variable dose method for treating idiopathic inflammatory bowel disease in a human subject in need thereof, comprising subcutaneously administering to the human subject:
a first dose of 160 mg of adalimumab administered to the human subject within a day; and
a second dose of 80 mg of adalimumab administered to the human subject within a day, wherein the second dose is administered two weeks following administration of the first dose.
US91937871. A method for purifying a composition comprising adalimumab, the method comprising:
(a) contacting a cation exchange adsorbent with a composition comprising adalimumab and adalimumab comprising one or more methylglyoxal (MGO)-modified arginine amino acids at position 30 (R30) of SEQ ID NO. 1, position 93 (R93) of SEQ ID NO. 1, position 108 (R108) of SEQ ID NO. 1, position 16 (R16) of SEQ ID NO. 2, position 259 (R259) of SEQ ID NO. 2, position 359 (R359) of SEQ ID NO. 2, or position 420 (R420) of SEQ ID NO. 2;
(b) removing adalimumab comprising one or more methylglyoxal (MGO)-modified arginine amino acids from the cation exchange adsorbent; and
(c) subsequently eluting the adalimumab from the cation exchange adsorbent using an elution buffer.
US92000691. A method of making a pharmaceutical composition, comprising mixing
(a) 25-100 mg of a low acidic species composition comprising adalimumab, wherein the low acidic species composition comprises less than 10% total acidic species of adalimumab,
wherein the acidic species of adalimumab have a net negative charge relative to the adalimumab main species and the acidic species comprise species selected from the group consisting of charge variants, structure variants, fragmentation variants and any combinations thereof; wherein the acidic species of adalimumab do not include process-related impurities selected from the group consisting of host cell proteins, host cell nucleic acids, chromatographic materials and media components, and wherein said low acidic species adalimumab composition demonstrates increased cartilage penetration as compared to a non-low acidic species composition of adalimumab; and
(b) a pharmaceutically acceptable carrier, thereby making a pharmaceutical composition.
US92000701. A low acidic species adalimumab pharmaceutical composition suitable for administration to a human subject comprising
40 mg of adalimumab wherein the low acidic species adalimumab pharmaceutical composition comprises less than 10% total acidic species of adalimumab, wherein the acidic species of adalimumab have a net negative charge relative to the adalimumab main species and the acidic species comprise species selected from the group consisting of charge variants, structure variants, fragmentation variants and any combinations thereof; wherein the acidic species of adalimumab do not include process-related impurities selected from the group consisting of host cell proteins, host cell nucleic acids, chromatographic materials and media components; and wherein said low acidic species adalimumab pharmaceutical composition demonstrates increased cartilage penetration as compared to a non-low acidic species composition of adalimumab; and
a pharmaceutically acceptable carrier.
US92063901. A method for controlling the oligosaccharide distribution of a recombinantly-expressed immunoglobulin comprising
supplementing during a production stage a cell culture media used in the recombinant expression of said immunoglobulin with a yeast hydrolysate supplement and/or a plant hydrolysate supplement to achieve a yeast hydrolysate concentration in the media of at least 11 g/L and/or a plant hydrolysate concentration of at least 7 g/L and
assessing the oligosaccharide distribution of the recombinantly-expressed immunoglobulin, thereby controlling the oligosaccharide distribution of the recombinantly-expressed immunoglobulin,
wherein the level of agalactosyl fucosylated biantennary oligosaccharides (sum of NGA2F and NGA2F-G1cNac) present on the recombinantly-expressed immunoglobulin is decreased as compared to the level of agalactosyl fucosylated biantennary oligosaccharides (sum of NGA2F and NGA2F-G1cNac) of the immunoglobulin recombinantly-expressed in cell culture media which is not supplemented with said yeast hydrolysate supplement and/or said plant hydrolysate supplement during the production stage; and/or
wherein the level of galactose containing fucosylated biantennary oligossacharides (sum of NA1F and NA2F) present on the recombinantly-expressed immunoglobulin is increased as compared to the level of galactose containing fucosylated biantennary oligossacharides (sum of NA1F and NA2F) of the immunoglobulin recombinantly-expressed in cell culture media which is not supplemented with said yeast hydrolysate supplement and/or said plant hydrolysate supplement during the production stage.
US92207811. A stable liquid aqueous pharmaceutical formulation comprising
(a) a human IgG1 anti-human Tumor Necrosis Factor alpha (TNFα) antibody at a concentration of 50 mg/ml,
(b) a polyol,
(c) a polysorbate, and
(d) a buffer system comprising an organic acid and having a pH of 4 to 8,
wherein the antibody is D2E7, and
wherein the formulation is suitable for subcutaneous injection.
US92340321. A fed-batch method for producing adalimumab in mammalian cells in culture, wherein said mammalian cells express said adalimumab, the method comprising:
culturing the mammalian cells in a cell culture production medium in large scale at a first temperature; and
then reducing the temperature under which the mammalian cells are cultured to a second lower temperature during adalimumab production, wherein said adalimumab is produced at a titer of at least 1.3 g/L in said cell culture and said adalimumab is further purified by a process including Protein A affinity chromatography.
US92340331. A process for producing a recombinantly-expressed immunoglobulin comprising a heavy chain variable region comprising the sequence of SEQ ID NO: 2 and a light chain variable region comprising the sequence of SEQ ID NO: 7, comprising
culturing a mammalian cell which recombinantly expresses the immunoglobulin in a cell culture media comprising a yeast hydrolysate and/or a plant hydrolysate, thereby producing the recombinantly-expressed immunoglobulin,
wherein the level of agalactosyl fucosylated biantennary oligosaccharides (sum of NGA2F and NGA2F-GlcNac) present on the produced immunoglobulin is decreased as compared to the level of agalactosyl fucosylated biantennary oligosaccharides (sum of NGA2F and NGA2F-GlcNac) of immunoglobulin produced in cell culture media which does not comprise said yeast hydrolysate, and/or said plant hydrolysate; and/or wherein the level of galactose containing fucosylated biantennary oligosaccharides (sum of NA1F and NA2F) present on the produced immunoglobulin is increased as compared to the level of galactose containing fucosylated biantennary oligosaccharides (sum of NA1F and NA2F) of immunoglobulin produced in cell culture media which does not comprise said yeast hydrolysate and/or said plant hydrolysate; and
wherein the level of agalactosyl fucosylated biantennary oligosaccharides (sum of NGA2F and NGA2F-GlcNAc) present on the produced immunoglobulin is 66%-69%; and/or wherein the level of fucosylated biantennary oligosaccharides (sum of NA1F and NA2F) present on the produced immunoglobulin is 29%-31%.
US92491821. A method for producing a preparation comprising a protein of interest and a reduced amount of at least one impurity, said method comprising:
(a) contacting a sample mixture comprising the protein of interest and the at least one impurity to a hydrophobic interaction chromatography (HIC) media in the presence of a load buffer and collecting a flow through fraction, such that the protein of interest binds to the HIC media at a Kp of at least 90; and
(b) contacting said hydrophobic interaction chromatography media with a wash buffer solution having a salt concentration within 20% of the salt concentration of the load buffer and collecting a wash fraction;
wherein the flow through and/or wash fractions constitute a preparation comprising a protein of interest and having a reduced amount of the impurity relative to the sample mixture.
US92551431. A composition comprising adalimumab, wherein more than 25% of the total N-linked oligosaccharides present on said adalimumab are of a galactose-containing fucosylated biantennary oligosaccharide form (sum of NA1F+NA2F).
US92669491. A method for producing a composition comprising an immunoglobulin comprising the 6 CDR domains of adalimumab, the method comprising:
culturing a mammalian cell producing an immunoglobulin comprising the 6 CDR domains of adalimumab in a cell culture media comprising 2 g/L to 11 g/L of each of one or more basic amino acids selected from the group consisting of arginine, lysine, ornithine and histidine, and combinations thereof, to produce a composition comprising an immunoglobulin comprising the 6 CDR domains of adalimumab, wherein the composition comprises less than 20% total acidic species of the immunoglobulin, and
wherein the acidic species of the immunoglobulin correspond to the peaks that elute earlier than the main peak in a WCX-10 HPLC chromatogram of the immunoglobulin, and wherein the WCX-10 HPLC chromatogram is generated using a first mobile phase of 10 mM Sodium Phosphate dibasic (pH 7.5) and a second mobile phase of 10 mM Sodium Phosphate dibasic, 500 mM Sodium Chloride (pH 5.5), and wherein the WCX-10 HPLC chromatogram is generated using detection at 280 nm.
US92720411. A stable liquid aqueous pharmaceutical formulation comprising
(a) a human IgG1 anti-human Tumor Necrosis Factor alpha (TNFα) antibody at a concentration of 50 mg/ml,
(b) a polyol,
(c) a polysorbate, and
(d) a buffer system comprising acetate and having a pH of 4 to 8,
wherein the antibody is D2E7, and
wherein the formulation is suitable for subcutaneous injection.
US92731321. A method of treating a disorder in which TNFα activity is detrimental in a subject, the method comprising administering a liquid pharmaceutical composition comprising a therapeutically effective amount of adalimumab and a pharmaceutically acceptable carrier to the subject such that the disorder is treated,
wherein the adalimumab is produced in a Chinese Hamster Ovary (CHO) cell expression system;
wherein the disorder is selected from the group consisting of rheumatoid arthritis, Crohn's disease, ulcerative colitis, ankylosing spondylitis, psoriatic arthritis, psoriasis, hidradenitis suppurativa, and juvenile rheumatoid arthritis; and
wherein the composition is characterized in that when the composition is assayed in a cathepsin L kinetic assay, a level of cathepsin L activity less than 1.84 RFU/s/mg of adalimumab is observed, wherein the cathepsin L kinetic assay comprises:
i) diluting the composition in a polystyrene container in a solution containing 25 mM NaOAc, 5 mM DTT and 1 mM EDTA at pH 5.5,
ii) adding dextran sulfate to a concentration of 0.035 μg/mL and incubating at 37° C. for six hours,
iii) adding Z-leucine-arginine covalently bound at its C-terminus to a fluorescent 7-amino-4-methyl coumarin (Z-leucine-arginine-AMC), wherein the diluting, adding, and incubating steps are sufficient to permit the measurement of cathepsin L hydrolysis of the Z-leucine-arginine-AMC within a linear range, and
iv) measuring Z-leucine-arginine-AMC hydrolysis in the linear range in RFU/s/mg of adalimumab.
US92843701. A method for treating polyarticular juvenile idiopathic arthritis in a subject comprising subcutaneously administering adalimumab to the subject every other week, wherein 20 mg of adalimumab is administered to the subject every other week if the subject weighs at least 15 kg and less than 30 kg, or wherein 40 mg of adalimumab is administered to the subject every other week if the subject weighs more than or equal to 30 kg.
US92843711. A method of producing adalimumab, comprising culturing in large-scale mammalian cells that express adalimumab in a cell culture production medium, wherein the pH of the cell culture production medium is adjusted from a first pH to a second pH during said culturing, the second pH being lower than the first pH, and the cell culture production medium is not removed but is supplemented with glucose or one or more other nutrients at least once during adalimumab production; and wherein adalimumab produced by the mammalian cells is purified from the cell culture production medium using a process including Protein A affinity purification.
US92905681. A process for producing a recombinantly-expressed immunoglobulin comprising a heavy chain variable region comprising the sequence of SEQ ID NO: 2 and a light chain variable region comprising the sequence of SEQ ID NO: 7, comprising culturing a mammalian cell which recombinantly expresses the immunoglobulin during a production stage in a cell culture media comprising at least 0.4 g/L of asparagine, thereby producing the recombinantly-expressed immunoglobulin,
wherein the level of agalactosyl fucosylated biantennary oligosaccharides (sum of NGA2F and NGA2F-GlcNac) present on the produced immunoglobulin is increased as compared to the level of agalactosyl fucosylated biantennary oligosaccharides (sum of NGA2F and NGA2F-GlcNac) of immunoglobulin produced in cell culture media which does not comprise said asparagine during the production stage; and/or wherein the level of galactose containing fucosylated biantennary oligossacharides (sum of NA1F and NA2F) present on the produced immunoglobulin is decreased as compared to the level of galactose containing fucosylated biantennary oligossacharides (sum of NA1F and NA2F) of immunoglobulin produced in cell culture media which does not comprise said asparagine during the production stage.
US93020111. A stable liquid aqueous pharmaceutical formulation comprising
(a) 50 mg/ml of a human IgG1 anti-human Tumor Necrosis Factor alpha (TNFα) antibody, wherein the antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO:1
and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 2;
(b) a polyol;
(c) a polysorbate; and
(d) a buffer system comprising an organic acid;
wherein the formulation has a pH of 4 to 8, and wherein the formulation is suitable for subcutaneous injection.
US93155741. A method for producing a composition comprising an immunoglobulin comprising the 6 CDR domains of adalimumab, the method comprising:
contacting a first sample comprising the immunoglobulin, wherein the sample comprises more than 10% total acidic species of the immunoglobulin, to a first chromatography media in the presence of a loading buffer to produce a first chromatography sample,
wherein the first chromatography media is selected from the group consisting of an ion exchange chromatography media, an affinity chromatography media and a hydrophobic interaction chromatography (HIC) media,
wherein the first chromatography sample comprises a composition of the immunoglobulin comprising less than 10% total acidic species of the immunoglobulin, wherein the acidic species of the immunoglobulin correspond to the peaks that elute earlier than the main peak in a WCX-10 HPLC chromatogram of the immunoglobulin, wherein the WCX-10 HPLC chromatogram is generated using a first mobile phase of 10 mM Sodium Phosphate dibasic (pH 7.5) and a second mobile phase of 10 mM Sodium Phosphate dibasic, 500 mM Sodium Chloride (pH 5.5), and wherein the WCX-10 HPLC chromatogram is generated using detection at 280 nm.
US93281651. A liquid pharmaceutical composition comprising adalimumab and one or more of sorbitol, a sugar, glycerol, or a preservative,
wherein the adalimumab is expressed in a Chinese Hamster Ovary (CHO) cell expression system; and the composition is characterized in that when the composition is assayed in a cathepsin L kinetic assay, a level of cathepsin L activity no greater than 1.3 RFU/s/mg of adalimumab is observed, wherein the cathepsin L kinetic assay comprises:
i.) diluting the composition in a polystyrene container in a solution containing 25 mM NaOAc, 5 mM DTT and 1 mM EDTA at pH 5.5,
ii.) adding dextran sulfate to a concentration of 0.035 μg/mL and incubating at 37° C. for six hours,
iii.) adding Z-leucine-arginine covalently bound at its C-terminus to a fluorescent 7-amino-4-methyl coumarin (Z-leucine-arginine-AMC), wherein the diluting, adding, and incubating steps are sufficient to permit the measurement of cathepsin L hydrolysis of the Z-leucine-arginine-AMC within a linear range, and
iv.) measuring Z-leucine-arginine-AMC hydrolysis in the linear range in RFU/s/mg of adalimumab.
US93343191. A composition comprising adalimumab, wherein the composition comprises less than 10% total acidic species of adalimumab, wherein the acidic species of adalimumab correspond to the peaks that elute earlier than the main peak in a WCX-10 HPLC chromatogram of adalimumab, wherein the WCX-10 HPLC chromatogram is generated using a first mobile phase of 10 mM Sodium Phosphate dibasic (pH 7.5) and a second mobile phase of 10 mM Sodium Phosphate dibasic, 500 mM Sodium Chloride (pH 5.5), and wherein the WCX-10 HPLC chromatogram is generated using detection at 280 nm.
US93468791. A method for producing a composition comprising adalimumab, the method comprising:
contacting a first sample comprising adalimumab comprising more than 10% total acidic species of adalimumab to a first chromatography media in the presence of a loading buffer, wherein the first chromatography media is selected from the group consisting of an ion exchange chromatography media, an affinity chromatography media and a hydrophobic interaction chromatography (HIC) media, and
collecting a first chromatography sample, wherein the first chromatography sample comprises a composition of adalimumab comprising less than 10% total acidic species of adalimumab, wherein the acidic species of adalimumab correspond to the peaks that elute earlier than the main peak in a WCX-10 HPLC chromatogram of adalimumab, wherein the WCX-10 HPLC chromatogram is generated using a first mobile phase of 10 mM Sodium Phosphate dibasic (pH 7.5) and a second mobile phase of 10 mM Sodium Phosphate dibasic, 500 mM Sodium Chloride (pH 5.5), and wherein the WCX-10 HPLC chromatogram is generated using detection at 280 nm.
US93594341. A method for producing a composition comprising adalimumab, the method comprising culturing a mammalian cell producing adalimumab in cell culture media comprising 2 g/L to 11 g/L of each of one or more basic amino acids, to produce a composition comprising adalimumab,
wherein the composition comprises less than 20% total acidic species of adalimumab, wherein the acidic species of adalimumab correspond to the peaks that elute earlier than the main peak in a WCX-10 HPLC chromatogram of adalimumab, and wherein the WCX-10 HPLC chromatogram is generated using a first mobile phase of 10 mM Sodium Phosphate dibasic (pH 7.5) and a second mobile phase of 10 mM Sodium Phosphate dibasic, 500 mM Sodium Chloride (pH 5.5), and wherein the WCX-10 HPLC chromatogram is generated using detection at 280 nm.
US93656451. A composition comprising an antibody comprising the heavy and light chain variable domains of adalimumab, wherein less than 70% of the total N-linked oligosaccharides present on said antibody are of an agalactosyl fucosylated biantennary oligosaccharide form (sum NGA2F+NGA2F-GlcNAc).
US94996141. A method of producing a composition comprising an immunoglobulin with an increased level of mannosylated N-glycans and/or a decreased level of fucosylated N-glycans, wherein the immunoglobulin comprises a light chain variable region of SEQ ID NO:1 and a heavy chain variable region of SEQ ID NO:2, said method comprising: culturing a mammalian host cell expressing said immunoglobulin in cell culture media supplemented with 5 mM-100 mM sucrose, thereby producing said composition comprising said immunoglobulin with an increased level of mannosylated N-glycans and/or a decreased level of fucosylated N-glycans as compared to a control, wherein said control is a composition comprising the immunoglobulin produced by culturing the mammalian host cell expressing said immunoglobulin in cell culture media which is not supplemented with sucrose.
US95058341. A method for increasing the galactosylation level of a recombinantly expressed antibody comprising the heavy and light chain variable domains of adalimumab, comprising supplementing a chemically defined (CD) cell culture media used in the expression of the antibody with a manganese supplement, thereby increasing the galactosylation level of the antibody, wherein the galactosylation level of the antibody is increased as compared to the galactosylation level of the antibody recombinantly expressed in the chemically defined cell culture media which is not supplemented with the manganese supplement.
US95122141. A method for controlling the oligosaccharide distribution of a recombinantly-expressed immunoglobulin comprising a heavy chain variable region comprising the sequence of SEQ ID NO: 2 and a light chain variable region comprising the sequence of SEQ ID NO: 7, comprising
supplementing during a production stage a cell culture medium used in the recombinant expression of said immunoglobulin with a yeast hydrolysate and/or a plant hydrolysate to achieve a yeast hydrolysate concentration in the medium of at least 11 g/L and/or a plant hydrolysate concentration of at least 7 g/L; and
assessing the oligosaccharide distribution of the recombinantly-expressed immunoglobulin, thereby controlling the oligosaccharide distribution of the recombinantly-expressed immunoglobulin,
wherein the level of agalactosyl fucosylated biantennary oligosaccharides (sum of NGA2F and NGA2F-GlcNac) present on the recombinantly-expressed immunoglobulin is decreased as compared to the level of agalactosyl fucosylated biantennary oligosaccharides (sum of NGA2F and NGA2F-GlcNac) of the immunoglobulin recombinantly-expressed in cell culture medium which is not supplemented with said yeast hydrolysate supplement and/or said plant hydrolysate supplement during the production stage; and/or
wherein the level of galactose containing fucosylated biantennary oligosaccharides (sum of NA1F and NA2F) present on the recombinantly-expressed immunoglobulin is increased as compared to the level of galactose containing fucosylated biantennary oligosaccharides (sum of NA1F and NA2F) of the immunoglobulin recombinantly-expressed in cell culture medium which is not supplemented with said yeast hydrolysate supplement and/or said plant hydrolysate supplement during the production stage.
US95122161. A method for treating moderate to severe chronic plaque psoriasis, comprising subcutaneously administering to an adult patient having moderate to severe chronic plaque psoriasis an initial dose of 80 mg of adalimumab, followed by 40 mg of adalimumab every other week starting one week after said first dosing, wherein the patient achieves at least Psoriasis Area and Severity Index (PASI) 75 response at week 12 of the treatment.
US95229531. A pre-filled syringe comprising a low acidic species composition comprising adalimumab, wherein the composition comprises less than 10% total acidic species of adalimumab, wherein the acidic species of adalimumab have a net negative charge relative to the adalimumab main species and the acidic species comprise species selected from the group consisting of charge variants, structure variants, fragmentation variants and any combinations thereof,
wherein the acidic species of adalimumab do not include process-related impurities selected from the group consisting of host cell proteins, host cell nucleic acids, chromatographic materials and media components, and
wherein the % acidic species is determined using WCX-10 HPLC wherein the WCX-10 HPLC chromatogram is generated using a first mobile phase of 10 mM Sodium Phosphate dibasic (pH 7.5) and a second mobile phase of 10 mM Sodium Phosphate dibasic, 500 mM Sodium Chloride (pH 5.5) and wherein the WCX-10 HPLC chromatogram is generated using detection at 280 nm.
US95462121. A method for treating rheumatoid arthritis in a human subject, comprising administering subcutaneously to a human subject having rheumatoid arthritis a total body dose of 40 mg of a human anti-TNFα antibody once every 13-15 days for a time period sufficient to treat the rheumatoid arthritis, wherein the anti-TNFα antibody comprises: an IgG1 heavy chain constant region; a variable light (“VL”) chain region comprising a CDR1 having the amino acid sequence of SEQ ID NO:7, a CDR2 having the amino acid sequence of SEQ ID NO:5, and a CDR3 having the amino acid sequence of SEQ ID NO:3; and a variable heavy (“VH”) chain region comprising a CDR1 having the amino acid sequence of SEQ ID NO:8, a CDR2 having the amino acid sequence of SEQ ID NO:6 and a CDR3 having the amino acid sequence of SEQ ID NO:4, wherein the human subject achieves an ACR20.
US96242951. A method of treating a subject having oligoarthritis with a tender joint count (TJC)<5 or a swollen joint count (SJC)<5, in association with psoriatic arthritis, the method comprising administering to the subject about 40 mg of adalimumab once subcutaneously every other week,
wherein at least an ACR20 response is achieved following a treatment period of at least about 12 weeks and the at least an ACR20 response is maintained following a treatment period of at least about 48 weeks,
 thereby treating the subject having psoriatic arthritis with oligoarthritis in association with psoriatic arthritis.
US96690931. A method for treating polyarticular juvenile idiopathic arthritis in a subject comprising weighing the subject and subcutaneously administering adalimumab to the subject every other week, wherein 20 mg of adalimumab is administered to the subject every other week if the subject weighs at least 15 kg and less than 30 kg, or wherein 40 mg of adalimumab is administered to the subject every other week if the subject weighs more than or equal to 30 kg.
US96830331. A composition comprising adalimumab, wherein the composition comprises less than 10% total acidic species of adalimumab,
wherein the acidic species of adalimumab are product-related impurities and do not include process-related impurities comprising host cells and lysed host cells, and
wherein the acidic species of adalimumab correspond to the peaks that elute earlier than the main peak in a WCX-10 HPLC chromatogram of adalimumab as shown in  FIG. 141.

Friday, October 6, 2017

Amgen v. Sanofi

The Federal Circuit has not been kind to patents that claim antibodies by the structure of an antigen/epitope without specifying the antibody structure.  Yesterday, the Fed. Cir. found the jury instructions in Amgen v. Sanofi  that "in the case of a claim to antibodies, the correlation between structure and function may also be satisfied by the disclosure of a newly characterized antigen" to be erroneous since "The essential problem with the jury instruction given in this case is that it effectively permitted the jury to dispense with the required finding of a written description of the invention."  In a previous case, the Fed. Cir. found the term “anti-CD20 antibody” to be limited to the particular epitope on CD20 that Biogen’s antibody bound.  Another Fed. Cir. case found the claim "a neutralizing isolated human antibody, or antigen-binding portion thereof that binds to human IL-12 . . ." to be invalid for written description.  The Federal Circuit’s decisions are in conflict with the USPTO’s written description guidelines that allow claiming an antibody to a specific antigen even if the specification does not disclose a specific antibody that binds to the particular antigen.

https://lnkd.in/gbr9ATG

And I was taking a break!