Highly concentrated bispecific antibody formulation

Abstract

Provided herein is a stable aqueous pharmaceutical composition comprising a highly concentrated formulation of a bispecific epidermal growth factor receptor (EGFR) / hepatocyte growth factor receptor (c-Met) antibody, and a method for preparing the same. Also provided herein is a method for treating cancer in a subject in need thereof by subcutaneously administering to the subject a stable aqueous pharmaceutical composition disclosed herein.

Description

[Technical field]

[0001] (CROSS REFERENCE TO RELATED APPLICATIONS) This application claims priority to U.S. Provisional Application No. 63 / 177,518, filed April 21, 2021, U.S. Provisional Application No. 63 / 180,690, filed April 28, 2021, and U.S. Provisional Application No. 63 / 309,230, filed February 11, 2022, the entire contents of which are incorporated herein by reference in their entireties.

[0002] (Reference to electronically submitted sequence listing) This application contains a Sequence Listing, having a size of 42 KB, that has been submitted electronically via EFS-Web as an ASCII formatted Sequence Listing with the filename "JBI6529WOPCT1SEQLIST.txt" created on April 7, 2022. The Sequence Listing submitted via EFS-Web is a part of the present specification and is incorporated herein by reference in its entirety.

[0003] FIELD OF THEINVENTION Disclosed are compositions of stable compositions comprising bispecific EGFR / c-Met antibodies for administration, including subcutaneous administration, as well as methods of using and formulating the stable compositions.

[0004] BACKGROUND OF THEINVENTION The role of both epidermal growth factor receptor (EGFR, ErbB1 or HER1) and hepatocyte growth factor receptor (c-Met) in cancer is well established, making these targets attractive for combination therapy. Both receptors signal through the same survival and anti-apoptotic pathways (ERK and AKT). Combination therapies targeting EGFR and c-Met or bispecific anti-EGFR / c-Met molecules are being tested in various clinical trials.

[0005] Although bispecific anti-EGFR / c-Met antibodies have shown promising results, there remains a need in the art for pharmaceutical compositions comprising such antibodies that are stable for extended periods at refrigerated (2-8° C.) and ambient temperatures while being optimally formulated for their mode of administration.

[0006] (Summary of the invention) Disclosed herein is a stable aqueous pharmaceutical composition comprising a particular formulation of a bispecific antibody.

[0007] In one aspect, provided herein is a stable aqueous pharmaceutical composition comprising a bispecific epidermal growth factor receptor (EGFR) / hepatocyte growth factor receptor (c-Met) antibody and hyaluronidase, wherein the antibody is a first heavy chain (HC1), comprising an HC1 variable region 1 (VH1) comprising the amino acid sequence of SEQ ID NO: 13; a first light chain (LC1) comprising a light chain variable region 1 (VL1) comprising the amino acid sequence of SEQ ID NO: 14; A second heavy chain (HC2) comprising an HC2 variable region 2 (VH2) comprising the amino acid sequence of SEQ ID NO: 15; A second light chain (LC2) comprising a light chain variable region 2 (VL2) comprising the amino acid sequence of SEQ ID NO: 16; the composition comprising from about 1,050 mg to about 2,240 mg of the bispecific EGFR / c-Met antibody, and from about 13,000 U to about 28,000 U of hyaluronidase.

[0008] In some embodiments, the composition comprises about 1,050 mg of a bispecific EGFR / c-Met antibody.

[0009] In some embodiments, the composition comprises about 1,400 mg of a bispecific EGFR / c-Met antibody.

[0010] In some embodiments, the composition comprises about 1,575 mg of the bispecific EGFR / c-Met antibody.

[0011] In some embodiments, the composition comprises about 1,600 mg of a bispecific EGFR / c-Met antibody.

[0012] In some embodiments, the composition comprises about 2,100 mg of a bispecific EGFR / c-Met antibody.

[0013] In some embodiments, the composition comprises about 2,240 mg of the bispecific EGFR / c-Met antibody.

[0014] In one aspect, provided herein is a stable aqueous pharmaceutical composition comprising: a) a bispecific epidermal growth factor receptor (EGFR) / hepatocyte growth factor receptor (c-Met) antibody at about 144 mg / mL to about 176 mg / mL, the bispecific antibody being a first heavy chain (HC1) comprising an HC1 variable region 1 (VH1); a first light chain (LC1) comprising a light chain variable region 1 (VL1); a second heavy chain (HC2) comprising HC2 variable region 2 (VH2); and a second light chain (LC2) comprising a light chain variable region 2 (VL2); Including, VH1 comprises the heavy chain complementarity determining region 1 (HCDR1), HCDR2, and HCDR3 amino acid sequences of SEQ ID NOs: 1, 2, and 3, respectively; VL1 comprises the light chain complementarity determining region 1 (LCDR1), LCDR2, and LCDR3 amino acid sequences of SEQ ID NOs: 4, 5, and 6, respectively; VH2 comprises the HCDR1, HCDR2, and HCDR3 amino acid sequences of SEQ ID NOs: 7, 8, and 9, respectively; and VL2 comprises the LCDR1, LCDR2, and LCDR3 amino acid sequences of SEQ ID NOs: 10, 11, and 12, respectively; A bispecific antibody; b) about 10 mM to about 50 mM acetic acid and / or a pharma- ceutically acceptable acetate salt; c) about 6.8% (w / v) to about 10.2% (w / v) sucrose; d) about 0.036% (w / v) to about 0.084% (w / v) polysorbate 80 (PS80); e) about 0.8 mg / mL to about 1.2 mg / mL of methionine; f) about 16 μg / mL to about 24 μg / mL ethylenediaminetetraacetic acid (EDTA); g) optionally, about 1,000 U / mL to about 3,000 U / mL of hyaluronidase; h) a pH of about 5.2 to about 6.2; A stable aqueous pharmaceutical composition comprising:

[0015] In some embodiments, the stable aqueous pharmaceutical composition comprises about 160 mg / mL of a bispecific EGFR-cMet antibody, about 30 mM acetic acid and / or a pharma- ceutically acceptable acetate salt, about 8.5% sucrose, and about 1 mg / mL L-methionine, polysorbate 80 to a final concentration of about 0.06% (w / v), and EDTA to a final concentration of about 20 μg / mL, wherein the stable aqueous pharmaceutical composition has a pH of about 5.7, and wherein the bispecific EGFR-cMet antibody comprises heavy chain 1 (HC1) comprising the amino acid sequence of SEQ ID NO: 17, HC2 comprising the amino acid sequence of SEQ ID NO: 19, light chain 1 (LC1) comprising the amino acid sequence of SEQ ID NO: 18, and LC2 comprising the amino acid sequence of SEQ ID NO: 20.

[0016] In some embodiments, the stable aqueous pharmaceutical composition comprises about 160 mg / mL of a bispecific EGFR-cMet antibody, about 30 mM acetic acid and / or a pharma- ceutically acceptable acetate salt, about 8.5% sucrose, and about 1 mg / mL L-methionine, with polysorbate 80 at a final concentration of about 0.06% (w / v), EDTA at a final concentration of about 20 μg / mL, and rHuPH20 at a final concentration of about 2,000 U / mL, wherein the stable aqueous pharmaceutical composition has a pH of about 5.7, and wherein the bispecific EGFR-cMet antibody comprises heavy chain 1 (HC1) comprising the amino acid sequence of SEQ ID NO: 17, HC2 comprising the amino acid sequence of SEQ ID NO: 19, light chain 1 (LC1) comprising the amino acid sequence of SEQ ID NO: 18, and LC2 comprising the amino acid sequence of SEQ ID NO: 20.

[0017] Also provided herein is a method of treating cancer in a subject in need thereof, the method comprising administering to the subject a stable aqueous pharmaceutical composition disclosed herein.

[0018] Also provided herein is a method of reducing an infusion-related reaction in a subject treated with amivantamab, comprising subcutaneously administering to the subject a stable aqueous pharmaceutical formulation disclosed herein.

[0019] Also provided herein is a method for preparing a stable aqueous pharmaceutical composition of a bispecific antibody targeting EGFR and cMet, the bispecific antibody targeting EGFR and cMet comprising a first heavy chain (HC1) comprising HC1 variable region 1 (VH1), a first light chain (LC1) comprising light chain variable region 1 (VL1), a second heavy chain (HC2) comprising HC2 variable region 2 (VH2), and a second light chain (LC2) comprising light chain variable region 2 (VL2), wherein VH1 is selected from the group consisting of SEQ ID NOs: 1, 2, and 3, respectively. and 3, VL1 comprises heavy chain complementarity determining region 1 (LCDR1), LCDR2 and LCDR3 comprising the amino acid sequences of SEQ ID NOs: 4, 5 and 6, respectively, VH2 comprises HCDR1, HCDR2 and HCDR3 amino acid sequences of SEQ ID NOs: 7, 8 and 9, respectively, and VL2 comprises LCDR1, LCDR2 and LCDR3 amino acid sequences of SEQ ID NOs: 10, 11 and 12, respectively. The methods include combining a composition comprising about 160 mg / mL of a bispecific antibody, about 30 mM acetic acid and / or a pharma- ceutically acceptable acetate salt, about 8.5% sucrose, and about 1 mg / mL of L-methionine with polysorbate 80 to a final concentration of about 0.06% (w / v) and EDTA to a final concentration of about 20 μg / mL, optionally rHuPH20 to a final concentration of about 2,000 U / mL, wherein the stable aqueous pharmaceutical composition has a pH of about 5.7.

[0020] Also provided herein is a kit comprising the stable liquid pharmaceutical formulation disclosed herein and instructions for its use.

[0021] Further provided herein is an article of manufacture comprising a container holding the stable liquid pharmaceutical formulation disclosed herein. [Brief description of the drawings]

[0022] [Figure 1] 4 shows serum concentration-time profiles of amivantamab doses. [Diagram 2] Saturation of soluble free EGFR and MET after the first SC amivantamab dose is shown. LLOQ - lower limit of quantification, Pre - pre-dose, "C" - treatment cycle (each cycle was 28 days), "D" - days within treatment cycle.

[0023] DETAILED DESCRIPTION OF THE PRESENT EMBODIMENT The methods of the present disclosure may be more readily understood by reference to the following detailed description taken in conjunction with the accompanying drawings, which form a part of this disclosure: It is to be understood that the methods of the present disclosure are not limited to the specific methods described and / or illustrated herein, and further, the terminology used herein is for the purpose of describing particular embodiments by way of example only, and is not intended to be limiting to the methods claimed.

[0024] Unless otherwise stated, any description of possible mechanisms or modes of operation or reasons for improvement is intended to be illustrative only, and the compositions and methods of the present disclosure are not limited by the merits or demerits of any such proposed mechanisms or modes of operation or reasons for improvement.

[0025] When a range of numerical values ​​is recited or established herein, the range includes its endpoints, and all individual integers and rational numbers within the range, and also includes each of the narrower ranges formed by all the various possible combinations of these endpoints and internal integers and rational numbers, forming a subgroup of the larger group of values ​​within the recited range, as if each of the narrower ranges were explicitly recited. When a range of numerical values ​​is recited herein as being greater than the recited value, the range is nevertheless finite, and its upper limit is defined by a value that is operable within the context of the invention described herein. When a range of numerical values ​​is recited herein as being less than the recited value, the range is nevertheless defined by a lower limit by a non-zero value. It is not intended that the scope of the invention be limited to the specific values ​​recited in defining the range. All ranges are inclusive and combinable.

[0026] When values ​​are expressed as approximations, by use of the antecedent "about," it is understood that the particular value forms another embodiment. Reference to a particular numerical value is intended to include at least the particular value unless the context clearly indicates otherwise.

[0027] It is also understood that certain features of the disclosed compositions and methods that are, for clarity, described herein in the context of separate embodiments, may also be provided in combination in a single embodiment. Conversely, various features of the disclosed compositions and methods that are, for brevity, described in the context of a single embodiment, may also be provided separately or in any subcombination.

[0028] As used herein, the singular forms "a," "an," and "the" are intended to include plurals.

[0029] Various terms relating to the embodiments of the present specification are used throughout the specification and claims. Unless otherwise indicated, such terms are to be given their ordinary meaning in the art. Other specifically defined terms are to be interpreted in a manner consistent with the definitions provided herein.

[0030] As used herein, "about" is used when used in connection with a numerical range, cutoff, or specific value to indicate that the recited value may vary by up to 10% from the recited value. Since many of the numerical values ​​used herein are experimentally determined, those skilled in the art will understand that such determinations may, and often will, vary between different experiments. The values ​​used herein should not be considered unduly limited by this inherent variation. Thus, the term "about" is used to encompass a variation of ±10% or less, a variation of ±5% or less, a variation of ±1% or less, a variation of ±0.5% or less, or a variation of ±0.1% or less from the stated value.

[0031] Similarly, the term "comprising" is intended to include examples encompassed by the terms "consisting essentially of" and "consisting of." Similarly, the term "consisting essentially of" is intended to include examples encompassed by the term "consisting of."

[0032] The term "antibody" and similar terms are intended in a broad sense to include immunoglobulin molecules or fragments thereof, including monoclonal antibodies (such as murine, human, human-adapted, humanized, and chimeric monoclonal antibodies), antibody fragments, bispecific or multispecific antibodies, dimeric, tetrameric, or multimeric antibodies, and single-chain antibodies.

[0033] Immunoglobulins can be assigned to five major classes, namely IgA, IgD, IgE, IgG, and IgM, depending on the amino acid sequence of the heavy chain constant domain. IgA and IgG are further classified into isotypes IgA1, IgA2, IgG1, IgG2, IgG3, IgG4. Antibody light chains of any vertebrate species can be assigned to one of two clearly distinct types, kappa (κ) and lambda (λ), based on the amino acid sequence of their constant domain.

[0034] "Antibody fragment" refers to a portion of an immunoglobulin molecule that retains the antigen-binding properties of the parent full-length antibody. Exemplary antibody fragments are heavy chain complementarity determining regions (HCDRs) 1, 2, and 3, light chain complementarity determining regions (LCDRs) 1, 2, and 3, heavy chain variable region (VH), or light chain variable region (VL). Antibody fragments include Fab fragments, which are monovalent fragments consisting of the VL, VH, constant light chain (CL), and (constant heavy chain 1) CH1 domains, F(ab')2 fragments, which are bivalent fragments containing two Fab fragments linked by a disulfide bridge at the hinge region, Fd fragments consisting of the VH and CHI domains, Fv fragments consisting of the VL and VH domains of a single arm of an antibody, and domain antibody (dAb) fragments consisting of the VH domain (Ward et al., Nature, 341:544-546, 1989). The VH and VL domains can be engineered and linked together via synthetic linkers to form a variety of single chain antibody designs, where the VH / VL domains pair intramolecularly or, when the VH and VL domains are expressed by separate single chain antibody constructs, pair intermolecularly to form a monovalent antigen binding site, such as a single chain Fv (scFv) or diabody. For example, as described in WO 1998 / 44001, WO 1988 / 01649, WO 1994 / 13804, and WO 1992 / 01047. These antibody fragments are obtained using techniques known to those skilled in the art, and the fragments are screened for utility in the same manner as full length antibodies.

[0035] Antibody variable regions consist of a "framework" region interrupted by three "antigen-binding sites", which are defined using various terms: (i) the complementarity determining regions (CDRs), three in VH (HCDR1, HCDR2, HCDR3) and three in VL (LCDR1, LCDR2, LCDR3), are based on sequence diversity (Wu and Kabat J Exp Med 132:211-50, 1970; Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md., 1991), and (ii) the "hypervariable regions" ("HVRs" or "HVs"), three in VH (H1, H2, H3) and three in VL (L1, L2, L3), are based on sequence diversity (Wu and Kabat J Exp Med 132:211-50, 1970; Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md., 1991), which ... and (ii) the "hypervariable regions" ("HVRs" or "HVs"), three in VH (H1, H2, H3) and three in VL (L1, L2, 196:901-17,1987). Other terms include "IMGT-CDR" (Lefranc et al., Dev Comparat Immunol 27:55-77,2003) and "Specificity Determining Residue Usage" (SDRU) (Almagro Mol Recognit 17:132-43,2004). The International ImMunoGeneTics (IMGT) database (http: / / www_imgt_org) provides standard numbering and definitions for antigen binding sites. The correspondence between CDR, HV and IMGT descriptive outlines is provided in Lefranc et al., Dev Comparat Immunol 27:55-77,2003.

[0036] "Monoclonal antibody" refers to a preparation of antibody molecules of a single molecular composition. A monoclonal antibody composition exhibits a single binding specificity and affinity for a particular epitope, or, in the case of bispecific monoclonal antibodies, dual binding specificity for two distinct epitopes. Thus, a monoclonal antibody refers to an antibody population in which the amino acid composition of each heavy and each light chain is uniform, except for possible known modifications such as removal of the C-terminal lysine from the antibody heavy chain. Monoclonal antibodies may have heterogeneous glycosylation within the antibody population. Monoclonal antibodies may be monospecific or multispecific, or monovalent, bivalent, or multivalent. Bispecific antibodies are included in the term monoclonal antibody.

[0037] The term "biosimilar" (of an approved reference product / biological agent, i.e., a listed drug) refers to a biological product that is so similar to the reference product, with minor differences in clinically inactive ingredients, that there are no clinically important differences between the biosimilar and the reference product in terms of safety, purity, and potency, based on data from: (a) analytical studies that demonstrate that the biological product is so similar to the reference product, with minor differences in clinically inactive ingredients, (b) animal studies (including evaluation of toxicity), and / or (c) clinical trial(s) (including evaluation of immunogenicity and pharmacokinetics or pharmacodynamics) sufficient to demonstrate safety, purity, and potency under one or more appropriate conditions of use, such as the conditions of use for which the reference product is approved and the conditions for which the reference product is intended to be used, and the conditions for granting approval for the biosimilar. A biosimilar may be an interchangeable product that may substitute for the reference product in the pharmacy without the intervention of the prescribing healthcare professional. To comply with the further criteria of "interchangeability," a biosimilar is required to produce the same clinical outcome in any given patient as the reference product, and, when a biosimilar is administered multiple times to an individual, the risks of reduced safety or efficacy from alternating or switching between the use of the biosimilar and the reference product do not outweigh the risks of using the reference product without such alternation or switching. To the extent that the mechanism of the reference product is known, the biosimilar utilizes the same mechanism of action for the proposed conditions of use. The condition(s) of use defined, recommended, or proposed in the labeling proposed for the biosimilar are already approved for the reference product. The route of administration, dosage form, and / or strength of the biosimilar are the same as those of the reference product, and the biosimilar is manufactured, processed, packaged, or held in facilities that meet standards designed to ensure that the biosimilar continues to maintain its safety, purity, and strength. Biosimilars may contain minor modifications of the amino acid sequence (e.g., N- or C-terminal truncations) that are not expected to alter the performance of the biosimilar when compared to the reference product.

[0038] "Epitope" refers to a portion of an antigen to which an antibody specifically binds. Epitopes usually consist of chemically active (such as polar, non-polar, or hydrophobic) surface groups of moieties such as amino acids or polysaccharide side chains and may have specific three-dimensional structural characteristics as well as specific charge characteristics. Epitopes may be composed of contiguous and / or discontinuous amino acids that form a conformational spatial unit. In discontinuous epitopes, amino acids in different parts of the linear sequence of the antigen are brought into close proximity in three-dimensional space due to folding of the protein molecule.

[0039] "Hyaluronidase" refers to an enzyme that breaks down hyaluronic acid. This enzyme is often used to increase the distribution and absorption of other co-administered drugs into tissues (e.g., subcutaneous injections, subcutaneous infusions, such as hypodermoclysis, etc.). More specifically, human recombinant DNA-derived hyaluronidase enzyme PH20 (rHuPH20) is often used to increase drug infusion subcutaneously, especially when large volumes are injected.

[0040] "Variant" refers to a polypeptide or polynucleotide that differs from a reference polypeptide or polynucleotide by one or more modifications, such as, for example, a substitution, insertion, or deletion.

[0041] By "in combination with" it is meant that two or more therapeutic agents can be administered to a subject together in a mixture, simultaneously as single agents, or sequentially in any order as single agents.

[0042] "Treat", "treatment" and similar terms refer to both therapeutic and prophylactic or preventative treatments, including reducing the severity and / or frequency of a symptom, eliminating a symptom and / or the underlying cause of a symptom, reducing the frequency or likelihood of a symptom and / or the underlying cause of a symptom, ameliorating or repairing damage caused directly or indirectly by a malignancy. Treatment also includes prolonging survival as compared to the expected survival of a subject not receiving treatment. Subjects to be treated include those suffering from a condition or disease, as well as those susceptible to a condition or disease, or those in which a condition or disease is to be prevented.

[0043] "Therapeutically effective amount" refers to an amount of the disclosed combination therapy effective at the required dosage and for the required period of time to achieve the desired treatment. The therapeutically effective amount may vary depending on factors such as the subject's condition, age, sex, and weight, as well as the ability of the combination therapy to induce a desired response in the subject. Exemplary indicators of a therapeutically effective amount include, for example, an improvement in the patient's health, a reduction in tumor burden, the cessation or slowing of tumor growth, and / or the absence of metastasis of cancer cells to other locations in the body.

[0044] The term "cancer" as used herein is defined as a disease characterized by rapid and uncontrolled growth of abnormal cells. Cancer cells can spread locally or through the bloodstream and lymphatic system to other parts of the body. Examples of various cancers include, but are not limited to, solid malignant tumors, such as breast cancer (BC), prostate cancer, ovarian cancer (OC), cervical cancer, skin cancer, pancreatic cancer, gastroesophageal cancer (GEC), colorectal cancer (CRC), renal cell carcinoma (RCC), liver cancer, hepatocellular carcinoma (HCC), brain cancer, squamous cell carcinoma of the head and neck (SCCHN), lymphoma, leukemia, lung cancer (e.g., non-small cell lung cancer (NSCL) or small cell lung cancer (SCLC)), medullary thyroid carcinoma (MTC), and mesothelioma. Solid malignant tumors can be metastatic or unresectable. Solid malignant tumors can be histologically or cytologically confirmed.

[0045] Reference material (RM) as used herein refers to a vial containing an approximately 0.2 mL aliquot of amivantamab or its biosimilar and used in Good Manufacturing Practice (GMP) for clinical substances. The RM is stored at least at -60°C, thawed, and used as a control in amivantamab analytical assays.

[0046] A "subject" includes any human or non-human animal. A "non-human animal" includes any vertebrate, e.g., mammals and non-mammals, such as non-human primates, sheep, dogs, cats, horses, cows, chickens, amphibians, reptiles, etc. The terms "subject" and "patient" may be used interchangeably herein.

[0047] "Infusion-related reactions" or "IRR" can occur in some patients receiving bispecific EGFR / c-Met antibodies such as amivantamab. Signs and symptoms of IRR can include dyspnea, flushing, fever, chills, nausea, chest discomfort, hypotension, and / or vomiting. Systemic IRR, including severe reactions, can also occur upon introduction of a new protein therapeutic infusion.

[0048] explanation Disclosed herein is a stable aqueous pharmaceutical composition comprising a bispecific EGFR / c-Met antibody. The bispecific anti-EGFR / c-Met antibody may be provided in a suitable pharmaceutical composition comprising the bispecific anti-EGFR / c-Met antibody and a pharma- ceutically acceptable carrier. The stable aqueous pharmaceutical composition may comprise one or more diluents, adjuvants, excipients, or vehicles in which the bispecific anti-EGFR / c-Met antibody is administered subcutaneously. Exemplary excipients include one or more of a buffer, stabilizer, chelator, surfactant, and enzyme. According to some embodiments, the stable aqueous pharmaceutical composition comprising a bispecific EGFR / c-Met antibody further comprises a buffer, stabilizer, chelator, surfactant, and optionally hyaluronidase. The stable aqueous pharmaceutical composition comprising a bispecific EGFR / c-Met antibody provided herein may also be referred to as a pharmaceutical product or DP.

[0049] In some embodiments, the bispecific EGFR / c-Met antibody comprises a first heavy chain (HC1) comprising HC1 variable region 1 (VH1), a first light chain (LC1) comprising light chain variable region 1 (VL1), a second heavy chain (HC2) comprising HC2 variable region 2 (VH2), and a second light chain (LC2) comprising light chain variable region 2 (VL2), wherein VH1 comprises heavy chain complementarity determining region 1 (HCDR1), HCD1 and HCD2 variable region 2 (HCDR3) of SEQ ID NOs: 1, 2, and 3, respectively. VL1 comprises light chain complementarity determining region 1 (LCDR1), LCDR2 and HCDR3 amino acid sequences of SEQ ID NOs: 4, 5 and 6, respectively, VH2 comprises HCDR1, HCDR2 and HCDR3 amino acid sequences of SEQ ID NOs: 7, 8 and 9, respectively, and VL2 comprises LCDR1, LCDR2 and LCDR3 amino acid sequences of SEQ ID NOs: 10, 11 and 12, respectively (see Table 29).

[0050] In some embodiments, the first heavy chain (HC1) of the bispecific EGFR-cMet antibody comprises HC1 constant domain 3 (HC1 CH3) and HC1 variable region 1 (VH1). In some embodiments, the second heavy chain (HC2) of the bispecific EGFR-cMet antibody comprises HC2 constant domain 3 (HC2 CH3) and HC2 variable region 2 (VH2). In some embodiments, the first heavy chain (HC1) of the bispecific EGFR-cMet antibody comprises HC1 constant domain 3 (HC1 CH3) and HC1 variable region 1 (VH1, and the second heavy chain (HC2) of the bispecific EGFR-cMet antibody comprises HC2 constant domain 3 (HC2 CH3) and HC2 variable region 2 (VH2). In some embodiments, the first heavy chain (HC1) of the bispecific EGFR-cMet antibody comprises HC1 constant domain 2 and constant domain 3 (HC1 CH2-CH3) and HC1 variable region 1 (VH1). In some embodiments, the second heavy chain (HC2) of the bispecific EGFR-cMet antibody comprises HC2 constant domain 2 and constant domain 3 (HC2 CH2-CH3) and HC2 variable region 2 (VH2). In some embodiments, the first heavy chain (HC1) of the bispecific EGFR-cMet antibody comprises HC1 constant domain 2 and constant domain 3 (HC1 The second heavy chain (HC2) of the bispecific EGFR-cMet antibody comprises HC2 constant domain 2 and constant domain 3 (HC2 CH2-CH3) and HC2 variable domain 2 (VH2).

[0051] In some embodiments, the bispecific antibody comprises an asymmetric stabilizing mutation in the HC1 CH2-CH3 region, the HC2 CH2-CH3 region, or both. An "asymmetric stabilizing mutation" refers to a mutation in the first CH2-CH3 region and the second CH2-CH3 region that is at a different position in the first and second CH2-CH3 region and that favors (e.g., stabilizes) heterodimer formation between the first CH2-CH3 region and the second CH2-CH3 region over homodimer formation between the first CH2-CH3 region or the second CH2-CH3 region. Exemplary asymmetric stabilizing mutations in the HC1 CH2-CH3 region and the HC2 CH2-CH3 region, or in the HC2 CH2-CH3 region and the HC1 CH2-CH3 region, are as follows (residue numbering according to the EU index): F405L and K409R, respectively. Wild type and F405L / R409K, respectively. T366W and T366S / L368A / Y407V, respectively. T366Y / F405A and T394W / Y407T, respectively. T366W / F405W and T394S / Y407A, respectively. F405W / Y407A and T366W / T394S, respectively. L351Y / F405A / Y407V and T394W, respectively. T366I / K392M / T394W and F405A / Y407V, respectively. T366L / K392M / T394W and F405A / Y407V, respectively. L351Y / Y407A and T366A / K409F, respectively. L351Y / Y407A and T366V / K409F, respectively. Y407A and T366A / K409F, respectively. D399K / E356K and K409D / K392D, respectively, or D399K / E356K / E357K and K409D / K392D / K370 respectively.

[0052] In some embodiments, the bispecific EGFR-cMet antibody comprises an HC1 variable region comprising the amino acid sequence of SEQ ID NO: 13 and an LC1 variable region comprising the amino acid sequence of SEQ ID NO: 14 (see Table 29). In some embodiments, the bispecific antibody comprises an asymmetric stabilizing mutation in the HC1 CH2-CH3 region, the HC2 CH2-CH3 region, or both. In some embodiments, the bispecific antibody comprises K409R in the c-Met binding arm and F405L in the EGFR binding arm.

[0053] In some embodiments, the bispecific EGFR-cMet antibody comprises an HC2 variable region comprising the amino acid sequence of SEQ ID NO: 15 and an LC2 variable region comprising the amino acid sequence of SEQ ID NO: 16 (see Table 29).

[0054] In some embodiments, heavy chain 1 (HC1) comprises the amino acid sequence of SEQ ID NO: 17 and HC2 comprises the amino acid sequence of SEQ ID NO: 19 (see Table 29).

[0055] In some embodiments, light chain 1 (LC1) comprises the amino acid sequence of SEQ ID NO: 18, and LC2 comprises the amino acid sequence of SEQ ID NO: 20 (see Table 29).

[0056] In some embodiments, the bispecific EGFR-cMet antibody is amivantamab or a biosimilar thereof.

[0057] Amivantamab Amivantamab (JNJ-61186372) is a low-fucose fully human IgG1-based bispecific antibody against the EGFR and cMET tyrosine kinase receptors that has been approved by the FDA for patients with EGFR exon 20ins mutations after chemotherapy treatment.

[0058] Amivantamab is active against tumors with primary activating EGFR mutations (exon 19 deletions [exon19del], exon 21 leucine 858 to arginine substitutions ([L858R], and exon 20ins mutations), EGFR resistance mutations (tyrosine 790 to methionine [T790M], or cysteine ​​797 to serine [C797S] mutations), overexpression of wild-type EGFR, and activation of the cMet pathway.

[0059] Lazertinib is an oral, highly potent, mutation-selective, and irreversible, third-generation EGFR TKI that targets both exon 19del and exon 21 L858R EGFR activating mutations and the T790M resistance mutation. Lazertinib has demonstrated efficacy in participants with EGFR-mutated NSCLC, with activity observed in both systemic and central nervous system lesions, demonstrating its ability to cross the blood-brain barrier. Combining lazertinib, which targets the intracellular EGFR tyrosine kinase site, with amivantamab, which targets the extracellular EGFR ligand-binding domain, has the potential to more potently inhibit the EGFR signaling pathway, attenuate frequent EGFR-dependent and -independent resistance mechanisms to EGFR TKIs, and induce deeper responses than either agent alone.

[0060] According to some embodiments, the stable aqueous pharmaceutical composition comprises a bispecific EGFR-cMet antibody at about 100 mg / mL, 110 mg / mL, 120 mg / mL, 121 mg / mL, 122 mg / mL, 123 mg / mL, 124 mg / mL, 125 mg / mL, 126 mg / mL, 127 mg / mL, 128 mg / mL, 129 mg / mL, 130 mg / mL, 131 mg / mL, 132 mg / mL, 133 mg / mL, 134 mg / mL , 135mg / mL, 136mg / mL, 137mg / mL, 138mg / mL, 139mg / mL, 140mg / mL, 141mg / mL, 142mg / mL, 143mg / mL, 144mg / mL, 145mg / mL mL, 146mg / mL, 147mg / mL, 148mg / mL, 150mg / mL, 151mg / mL, 152mg / mL, 153mg / mL, 154mg / mL, 155mg / mL, 156mg / mL, 157mg / mL, 158mg / mL, 159mg / mL, 160mg / mL, 161mg / mL, 162mg / mL, 163mg / mL, 164mg / mL, 165mg / mL, 166mg / mL, 167mg / mL, 168 mg / mL, 169 mg / mL, 170 mg / mL, 171 mg / mL, 172 mg / mL, 173 mg / mL, 174 mg / mL, 175 mg / mL, 176 mg / mL, 177 mg / mL, 178 mg / mL, 17 In some embodiments, the bispecific EGFR-cMet antibody has a concentration of about 160 mg / mL.

[0061] In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered at a dose of about 140 mg to about 1750 mg. In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered at a dose of about 140 mg to about 2100 mg. In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered at a dose of about 1050 mg to about 2240 mg.

[0062] In some embodiments, the bispecific anti-EGFR / c-Met antibody is about 200 mg, about 210 mg, about 220 mg, about 230 mg, about 240 mg, about 250 mg, about 260 mg, about 270 mg, about 280 mg, about 290 mg, about 300 mg, about 310 mg, about 320 mg, about 330 mg, about 340 mg, about 350 mg, about 360 mg, about 370 mg, about 380 mg, about 390 mg, about 400 mg, about 410 mg, about 420 mg, about 430 mg, about 440 mg, about 450 mg, about 460 mg, about 470 mg, about 480 mg, about 490 mg, about 500 mg, about 510 mg, about 520 mg, about 530 mg, about 540 mg, about 550 mg, about 560 mg, about 570 mg, about 580 mg, about 590 mg, about 600 mg, about 610 mg, about 620 mg, about 630 mg, about 640 mg, about 650 mg, about 660 mg, about 670 mg, about 680 mg, about 690 mg, about 700 mg, about 710 mg, about 720 mg, about 730 mg, about 740 mg, about 750 mg, about 760 mg, about 770 mg, about 780 mg, about 790 mg, about 800 mg, about 810 mg, about 820 mg, about 830 mg, about 840 mg, about 850 mg, about 860 mg, about 870 mg, about 880 mg, about 890 mg, about 900 mg, about 910 mg, about 920 mg, about 930 mg, about 940 mg, about 950 mg, about 960 mg, about 970 mg, about 980 mg, about 990 mg, about 1000 mg, about 1010 mg, about 1020 mg, about 1030 mg, about 1040 mg, about 1050 mg, about 1060 mg, about 1070 mg, about 1080 mg, about 1090 mg, about 1100 mg, about 1110 mg, about 1120 mg, about 1130 mg, about 1140 mg, about 1150 mg, about 1160 mg, about 1170 mg, about 1180 mg, about 1190 mg, about 1200 mg, about 1210 mg, about 1220 mg, about 1230 mg, about 1240 mg, about 1250 mg, about 1260 mg, about 1270 mg, about 1280 mg, about 1290 mg, about 1300 mg, about 1310 mg, about 1320 mg, about 1330 mg, about 1340 mg, about 1350 mg, about 1360 mg, about 1370 mg, about 1380 mg, about 1390 mg, about 1400 mg, about 1410 mg, about 1420 mg, about 1430 mg, about 1440 mg, about 1450 mg, about 1460 mg, about 1470 mg, about 1480 mg, about 1490 mg, about 1500 mg,About 1510mg, about 1520mg, about 1530mg, about 1540mg, about 1550mg, about 1560mg, about 1570mg, about 1575mg, about 1580mg, about 159 0mg, about 1600mg, about 1610mg, 1620mg, about 1630mg, about 1640mg, about 1650mg, about 1660mg, about 1670mg, about 1680mg, about 1690mg, about 1700mg, about 1710mg, about 1720mg, about 1730mg, about 1740mg, about 1750mg, about 1760mg, about 1770mg, about 1780 mg, about 1790mg, about 1800mg, about 1810mg, about 1820mg, about 1830mg, about 1840mg, about 1850mg, about 1860mg, about 1870mg, about 1880mg, 1890mg, about 1900mg, about 1910mg, about 1920mg, about 1930mg, about 1940mg, about 1950mg, about 1960mg, about 1970m g, about 1980mg, about 1990mg, about 2000mg, about 2010mg, about 2020mg, about 2030mg, about 2040mg, about 2050mg, about 2060mg, about 2 The compound is administered in a dose of about 200 mg, about 2070 mg, about 2080 mg, about 2090 mg, about 2100 mg, about 2110 mg, about 2120 mg, about 2150 mg, about 2200 mg, about 2210 mg, about 2220 mg, about 2230 mg, about 2240 mg, about 2250 mg, about 2260 mg, about 2270 mg, about 2280 mg, about 2290 mg, or about 2300 mg.

[0063] In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered at a dose of about 350 mg, about 700 mg, about 1050 mg, about 1400 mg, about 1575 mg, or about 2100 mg. In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered at a dose of about 350 mg. In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered at a dose of about 700 mg. In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered at a dose of about 750 mg. In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered at a dose of about 800 mg. In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered at a dose of about 850 mg. In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered at a dose of about 900 mg. In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered at a dose of about 950 mg. In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered at a dose of about 1000 mg. In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered at a dose of about 1050 mg. In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered at a dose of about 1100 mg. In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered at a dose of about 1150 mg. In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered at a dose of about 1200 mg. In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered at a dose of about 1250 mg. In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered at a dose of about 1300 mg. In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered at a dose of about 1350 mg. In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered at a dose of about 1400 mg. In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered at a dose of about 1575 mg. In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered at a dose of about 1600 mg.In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered at a dose of about 2100 mg. In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered at a dose of about 2240 mg.

[0064] In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered once a week. In some embodiments, about 1050 mg of the bispecific anti-EGFR / c-Met antibody is administered once a week. In some embodiments, about 1400 mg of the bispecific anti-EGFR / c-Met antibody is administered once a week. In some embodiments, about 1575 mg of the bispecific anti-EGFR / c-Met antibody is administered once a week. In some embodiments, about 1600 mg of the bispecific anti-EGFR / c-Met antibody is administered once a week. In some embodiments, about 2100 mg of the bispecific anti-EGFR / c-Met antibody is administered once a week. In some embodiments, about 2240 mg of the bispecific anti-EGFR / c-Met antibody is administered once a week.

[0065] In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered once every two weeks. In some embodiments, about 1050 mg of the bispecific anti-EGFR / c-Met antibody is administered once every two weeks. In some embodiments, about 1400 mg of the bispecific anti-EGFR / c-Met antibody is administered once every two weeks. In some embodiments, about 1575 mg of the bispecific anti-EGFR / c-Met antibody is administered once every two weeks. In some embodiments, about 1600 mg of the bispecific anti-EGFR / c-Met antibody is administered once every two weeks. In some embodiments, about 2100 mg of the bispecific anti-EGFR / c-Met antibody is administered once every two weeks. In some embodiments, about 2240 mg of the bispecific anti-EGFR / c-Met antibody is administered once every two weeks.

[0066] In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered twice weekly. In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered once weekly. In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered once every two weeks. In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered once every three weeks. In some embodiments, the bispecific anti-EGFR / c-Met antibody is administered once every four weeks.

[0067] In the case of combination therapy, one or more anti-cancer agents may be administered using the recommended doses of the anti-cancer agents.

[0068] The buffering agent is suitable for adjusting the pH to about 5.0 to 6.2, such as pH 5.1 to 5.7. Exemplary buffers are histidine buffers or acetate buffers. According to some embodiments, the stable aqueous pharmaceutical composition comprises a buffer solution of about 8 mM, 9 mM, 10 mM, 11 mM, 12 mM, 13 mM, 14 mM, 15 mM, 16 mM, 17 mM, 18 mM, 19 mM, 20 mM, 21 mM, 22 mM, 23 mM, 24 mM, 25 mM, 26 mM, 27 mM, 28 mM, 29 mM, 30 mM, 31 mM, 32 mM, 33 mM, 34 mM, 35 mM, 36 mM, 37 mM, 38 mM, 39 mM, 40 mM, 41 mM, 42 mM, 43 mM, 44 mM, 45 mM, 46 mM, 47 mM, 48 mM, 49 mM, 50 mM, 51 mM, 52 mM, 53 mM, 54 mM, 55 mM, 56 mM, 57 mM, 58 mM, 59 mM, 60 mM, 61 mM, 62 mM, 63 mM, 64 mM, 65 mM, 66 mM, 67 mM, 68 mM, 69 mM, 70 mM, 71 mM, 72 mM, 73 mM, 74 mM, 75 mM, 76 mM, 77 mM, 78 mM, 79 mM, 80 mM, 81 mM, 82 mM, 83 mM, 84 mM, 85 mM, 86 mM, 87 mM, 88 mM, 89 mM, 90 mM, 91 mM, 92 mM, 93 mM, 94 mM, 95 mM, 96 mM, In some embodiments, the histidine and / or pharma- ceutically acceptable histidine salt has a concentration of about 10 mM. In some embodiments, the histidine and / or pharma- ceutically acceptable histidine salt has a concentration of about 50 mM. In further embodiments, the histidine and / or pharma- ceutically acceptable histidine salts include L-histidine and L-histidine hydrochloride monohydrate. According to some embodiments, the stable aqueous pharmaceutical composition comprises acetic acid and / or a pharma- ceutically acceptable acetate salt at a concentration of about 8 mM, 9 mM, 10 mM, 11 mM, 12 mM, 13 mM, 14 mM, 15 mM, 16 mM, 17 mM, 18 mM, 19 mM, 20 mM, 21 mM, 22 mM, 23 mM, 24 mM, 25 mM, 26 mM, 27 mM, 28 mM, 29 mM, 30 mM, 31 mM, 32 mM, 33 mM, 34 mM, 35 mM, 36 mM, 37 mM, 38 mM, 39 mM, 40 mM, 41 mM, 42 mM, 43 mM, 44 mM, 45 mM, 46 mM, 47 mM, 48 mM, 49 mM, or 50 mM. In some embodiments, the acetic acid and / or a pharma- ceutically acceptable acetate salt has a concentration of about 30 mM. In further embodiments, the acetic acid and / or a pharma- ceutically acceptable acetate salt comprises glacial acetic acid and / or sodium acetate trihydrate.

[0069] The stabilizer may include sucrose and optionally methionine. According to some embodiments, the stable aqueous pharmaceutical composition has a pH of about 6.0%, 6.1%, 6.2%, 6.3%, 6.4%, 6.5%, 6.6%, 6.7%, 6.8%, 6.9%, 7.0%, 7.1%, 7.2%, 7.3%, 7.4%, 7.5%, 7.6%, 7.7%, 7.8%, 7.9%, 8.0%, 8.1%, 8.2%, 8.3%, 8.4%, 8.5%, 8.6%, 8.7%, 8.8%, 8.9%, 9.0%, 9.1%, 9.2%, 9.3%, 9.4%, 9.5%, 9.6%, 9.7%, 9.8%, 9 ... In some embodiments, the stable aqueous pharmaceutical composition comprises sucrose at a concentration (percent weight to volume (% w / v)) of about 0.2%, 8.3%, 8.4%, 8.5%, 8.6%, 8.7%, 8.8%, 9.9%, 10.0%, 10.1%, 10.2%, 10.3%, 10.4%, 10.5%, 10.6%, 10.7%, 10.8%, 10.9%, or 11.0%. In some embodiments, the stable aqueous pharmaceutical composition comprises about 8.5% (w / v) sucrose. According to some aspects, the stable aqueous pharmaceutical composition comprises methionine (e.g., L-methionine) at a concentration of about 0.1 mg / mL, 0.2 mg / mL, 0.3 mg / mL, 0.4 mg / mL, 0.5 mg / mL, 0.6 mg / mL, 0.7 mg / mL, 0.8 mg / mL, 0.9 mg / mL, 1.0 mg / mL, 1.1 mg / mL, 1.2 mg / mL, 1.3 mg / mL, 1.4 mg / mL, 1.5 mg / mL, 1.6 mg / mL, 1.7 mg / mL, 1.8 mg / mL, 1.9 mg / mL, or 2.0 mg / mL. In one embodiment, the methionine comprises L-methionine and has a concentration of about 1.0 mg / mL.

[0070] A preferred surfactant is polysorbate 80 (PS80). According to some embodiments, the stable aqueous pharmaceutical composition comprises polysorbate 80 (PS80) at about 0.005%, 0.01%, 0.015%, 0.020%, 0.025%, 0.030%, 0.035%, 0.036%, 0.037%, 0.038%, 0.039%, 0.040%, 0.041%, 0.042%, 0.043%, 0.044%, 0.045%, 0.046%, 0.047%, 0.048%, 0.049%, 0.050%, 0.051%, 0.052%, 0.053%, 0.054%, 0.055%, 0.056%, 0.057%, 0.058%, 0.059%, 0.060%, 0.061%, 0.062%, 0.063%, 0.064%, 0.065%, 0.066%, 0.067%, 0.068%, 0.069%, 0.070%, 0.071%, 0.072%, 0.073%, 0.074%, 0.075%, 0.076%, 0.077%, 0.078%, 0.079%, 0.080%, 0.081%, 0.082%, 0.083%, 0.084%, 0.085%, 0.086%, 0.087%, 0.088%, 0.089%, 0.090%, 0.091%, 0.092%, 0.093%, 0.094%, 0.095%, 0.096%, 0.097%, 0.09 8%, 0.059%, 0.060%, 0.061%, 0.062%, 0.063%, 0.064%, 0.065%, 0.066%, 0.067%, 0.068%, 0.069%, 0.070%, 0.071%, 0.072%, 0.073%, 0.074%, 0.075%, 0. In some embodiments, the stable aqueous pharmaceutical composition comprises about 0.06% (w / v) PS80.

[0071] A preferred chelating agent is ethylenediaminetetraacetic acid (EDTA). According to some embodiments, the stable aqueous pharmaceutical composition comprises EDTA at a concentration of about 10 μg / mL, 11 μg / mL, 12 μg / mL, 13 μg / mL, 14 μg / mL, 15 μg / mL, 16 μg / mL, 17 μg / mL, 18 μg / mL, 19 μg / mL, 20 μg / mL, 21 μg / mL, 22 μg / mL, 23 μg / mL, 24 μg / mL, 25 μg / mL, 26 μg / mL, 27 μg / mL, 28 μg / mL, 29 μg / mL, or 30 μg / mL. In one embodiment, the EDTA has a concentration of about 20 μg / mL.

[0072] In some embodiments, the stable aqueous pharmaceutical composition comprising the bispecific EGFR / c-Met antibody comprises a hyaluronidase enzyme in an amount sufficient to increase dispersion of the antibody during subcutaneous administration. The hyaluronidase enzyme excipient according to the formulation of the present invention is characterized in that it does not adversely affect the molecular integrity of the bispecific EGFR / c-Met antibody in the stable pharmaceutical composition described herein. Furthermore, the hyaluronidase enzyme merely modifies the delivery of the bispecific EGFR / c-Met antibody to the systemic circulation, but does not have any properties that may provide or contribute to the therapeutic effect of a systemically absorbed bispecific EGFR / c-Met antibody. The hyaluronidase enzyme is not systemically bioavailable and does not adversely affect the molecular integrity of the bispecific EGFR / c-Met antibody at the recommended storage conditions of the stable pharmaceutical composition according to the present invention. Several suitable hyaluronidase enzymes according to the present invention are known. A preferred enzyme is a human hyaluronidase enzyme, such as soluble human PH20 hyaluronidase, preferably a recombinant human hyaluronidase enzyme product known as rHuPH20. The amino acid sequence of the soluble human PH20 hyaluronidase includes soluble human PH20, known as rHuPH20 and available under CAS Registry Number 757971-58-7. Soluble human PH20 hyaluronidiase is described in WO 2004 / 078140 and U.S. Patent No. 7,767,429, which are incorporated by reference in their entireties. In some embodiments, the soluble hyaluronidase includes those whose sequences are set forth in any of SEQ ID NOs: 21-25. The soluble PH20 hyaluronidase contains a signal sequence for transport in the cell when expressed in the cell. Thus, in some embodiments, the amino acid sequence of the soluble PH20 hyaluronidase comprises SEQ ID NO: 26. In some embodiments, the amino acid sequence of the soluble PH20 hyaluronidase comprises SEQ ID NO: 22, i.e., residues 36-482 of wild-type human hyaluronidase. In some embodiments, the amino acid sequence of the soluble PH20 hyaluronidase comprises SEQ ID NO: 23. In some embodiments, the amino acid sequence of the soluble PH20 hyaluronidase comprises SEQ ID NO: 24.In some embodiments, the amino acid sequence of the soluble PH20 hyaluronidase rHuPH20 comprises SEQ ID NO: 25. In some embodiments, the amino acid sequence of the soluble PH20 hyaluronidase comprises SEQ ID NO: 21. In some embodiments, the soluble PH20 hyaluronidase, when expressed in a cell, comprises a mixture of species that may include any one or more of SEQ ID NO: 21-25 in varying abundance. The average molecular weight is 61 kDa.

[0073] rHuPH20 generally refers to a composition produced upon expression in cells, such as CHO cells, of a nucleic acid encoding residues 36-482 of SEQ ID NO:26 linked to a native or heterologous signal sequence (residues 1-35 of SEQ ID NO:26). rHuPH20 is produced by expression of a nucleic acid molecule encoding amino acids 1-482 (as set forth in SEQ ID NO:26) in mammalian cells. Translational processing removes the 35 amino acid signal sequence. Heterogeneity is present at the C-terminus such that when produced in culture, the product designated rHuPH20 comprises a mixture of species that may include any one or more of polypeptides 36-480, 36-481, and 36-482 of SEQ ID NO:26, as well as several shorter polypeptides, in varying abundance. Typically, rHuPH20 is produced in cells, such as CHO cells, that promote correct N-glycosylation to retain activity, e.g., DG44 CHO cells.

[0074] In some embodiments, the stable aqueous pharmaceutical composition comprises rHuPH20 at a concentration of about 500 U / mL, about 750 U / mL, about 1,000 U / mL, about 1,250 U / mL, about 1,500 U / mL, about 1,750 U / mL, about 2,000 U / mL, about 2,250 U / mL, about 2,500 U / mL, about 2,750 U / mL, about 3,000 U / mL, about 3,250 U / mL, about 3,500 U / mL, about 3,750 U / mL, or about 4,000 U / mL. In some embodiments, the stable aqueous pharmaceutical composition comprises rHuPH20 at a concentration of about 2,000 U / mL. In some embodiments, the stable aqueous pharmaceutical composition comprises rHuPH20 at a concentration of about 1,500 U / mL. According to some embodiments, the stable aqueous pharmaceutical composition comprises rHuPH20 at a concentration of about 0.005 mg / mL, about 0.0075 mg / mL, about 0.01 mg / mL, about 0.0125 mg / mL, about 0.015 mg / mL, about 0.0175 mg / mL, about 0.02 mg / mL, about 0.0225 mg / mL, about 0.025 mg / mL, about 0.0275 mg / mL, about 0.03 mg / mL, about 0.0325 mg / mL, about 0.035 mg / mL, about 0.0375 mg / mL, or about 0.04 mg / mL. In some embodiments, the stable aqueous pharmaceutical composition comprises rHuPH20 at a concentration of about 0.02 mg / mL.

[0075] In some embodiments, the stable aqueous pharmaceutical composition comprises about 10,000 U, about 11,000 U, about 12,000 U, about 13,000 U, about 13,200 U, about 14,000 U, about 15,000 U, about 16,000 U, about 17,000 U, about 17,500 U, about 18,000 U, about 19,000 U, about 19,500 U, about 19,600 U, about 19,680 U, about 19,700 U, about The stable aqueous pharmaceutical composition may comprise rHuPH20 at a dose of about 20,000 U, about 21,000 U, about 22,000 U, about 23,000 U, about 24,000 U, about 25,000 U, about 26,000 U, about 26,260 U, about 26,500 U, about 26,600 U, about 26,680 U, about 26,700 U, about 27,000 U, 28,000 U, about 29,000 U, about 30,000 U, or any value therebetween. In some embodiments, the stable aqueous pharmaceutical composition comprises rHuPH20 at a dose of about 13,000 U. In some embodiments, the stable aqueous pharmaceutical composition comprises rHuPH20 at a dose of about 13,200 U. In some embodiments, the stable aqueous pharmaceutical composition comprises rHuPH20 at a dose of about 14,000 U. In some embodiments, the stable aqueous pharmaceutical composition comprises rHuPH20 at a dose of about 17,500 U. In some embodiments, the stable aqueous pharmaceutical composition comprises rHuPH20 at a dose of about 18,000 U. In some embodiments, the stable aqueous pharmaceutical composition comprises rHuPH20 at a dose of about 19,680 U. In some embodiments, the stable aqueous pharmaceutical composition comprises rHuPH20 at a dose of about 20,000 U. In some embodiments, the stable aqueous pharmaceutical composition comprises rHuPH20 at a dose of about 26,000 U. In some embodiments, the stable aqueous pharmaceutical composition comprises rHuPH20 at a dose of about 26,260 U. In some embodiments, the stable aqueous pharmaceutical composition comprises rHuPH20 at a dose of about 26,300 U. In some embodiments, the stable aqueous pharmaceutical composition comprises rHuPH20 at a dose of about 26,400 U. In some embodiments, the stable aqueous pharmaceutical composition comprises rHuPH20 at a dose of about 26,500 U. In some embodiments, the stable aqueous pharmaceutical composition comprises rHuPH20 at a dose of about 26,600 U. In some embodiments, the stable aqueous pharmaceutical composition comprises rHuPH20 at a dose of about 27,000 U.In some embodiments, the stable aqueous pharmaceutical composition comprises rHuPH20 at a dose of about 27,500 U. In some embodiments, the stable aqueous pharmaceutical composition comprises rHuPH20 at a dose of about 28,000 U. In some embodiments, the stable aqueous pharmaceutical composition comprises rHuPH20 at a dose of about 28,500 U. In some embodiments, the stable aqueous pharmaceutical composition comprises rHuPH20 at a dose of about 29,000 U. In some embodiments, the stable aqueous pharmaceutical composition comprises rHuPH20 at a dose of about 30,000 U.

[0076] In some embodiments, the bispecific EGFR / c-Met antibody stable aqueous pharmaceutical composition comprises histidine, and / or a pharma- ceutically acceptable histidine salt, sucrose, polysorbate 80 (PS80), methionine, EDTA, a pH of about 5.2 to about 6.2, and, optionally, hyaluronidase. The stable aqueous pharmaceutical composition may further comprise about 128 mg / mL to about 192 mg / mL of a bispecific EGFR-cMet antibody, about 10 mM to about 50 mM of histidine and / or a pharma- ceutically acceptable histidine salt, about 6.8% (w / v) to about 10.2% (w / v) of sucrose, about 0.036% (w / v) to about 0.084% (w / v) of polysorbate 80 (PS80), about 0.8 mg / mL to about 1.2 mg / mL of methionine, about 16 μg / mL to about 24 μg / mL of EDTA, and a pH of about 5.2 to about 6.2. The stable aqueous pharmaceutical composition may optionally comprise about 1,000 U / mL to about 3,000 U / mL of hyaluronidase. In some embodiments, the stable aqueous pharmaceutical composition comprises about 160 mg / mL of bispecific EGFR-cMet antibody, about 10 mM histidine and / or a pharma- ceutically acceptable histidine salt, about 8.5% (w / v) sucrose, about 0.06% (w / v) polysorbate 80 (PS80), about 1 mg / mL methionine, about 20 μg / mL EDTA, a pH of about 5.7, and, optionally, about 2,000 U / mL hyaluronidase.

[0077] In other embodiments, the bispecific EGFR / c-Met antibody stable aqueous pharmaceutical composition comprises acetic acid, and / or a pharma- ceutically acceptable acetate salt, sucrose, polysorbate 80 (PS80), methionine, EDTA, a pH of about 5.2 to about 6.2, and, optionally, hyaluronidase. The stable aqueous pharmaceutical composition may comprise about 128 mg / mL to about 192 mg / mL of a bispecific EGFR-cMet antibody, about 10 mM to about 50 mM of acetic acid and / or a pharma- ceutical acceptable acetate, about 6.8% (w / v) to about 10.2% (w / v) of sucrose, about 0.036% (w / v) to about 0.084% (w / v) of polysorbate 80 (PS80), about 0.8 mg / mL to about 1.2 mg / mL of methionine, about 16 μg / mL to about 24 μg / mL of EDTA, and a pH of about 5.2 to about 6.2. The stable aqueous pharmaceutical composition may optionally comprise about 1,000 U / mL to about 3,000 U / mL of hyaluronidase. In some embodiments, the stable aqueous pharmaceutical composition comprises about 160 mg / mL of a bispecific EGFR-cMet antibody, about 30 mM acetic acid and / or a pharma- ceutically acceptable acetate salt, about 8.5% (w / v) sucrose, about 0.06% (w / v) polysorbate 80 (PS80), about 1 mg / mL methionine, about 20 μg / mL EDTA, a pH of about 5.7, and, optionally, about 2,000 U / mL hyaluronidase.

[0078] Further provided herein is a method of treating cancer in a subject in need thereof by administering to the subject a therapeutically effective amount of a stable aqueous pharmaceutical composition of a bispecific EGFR / c-Met antibody disclosed herein. The cancer may be a solid malignant tumor, such as breast cancer (BC), prostate cancer, ovarian cancer (OC), cervical cancer, skin cancer, pancreatic cancer, gastroesophageal cancer (GEC), colorectal cancer (CRC), renal cell carcinoma (RCC), liver cancer, hepatocellular carcinoma (HCC), brain cancer, squamous cell carcinoma of the head and neck (SCCHN), lymphoma, leukemia, lung cancer (e.g., non-small cell lung cancer (NSCLC) or small cell lung cancer (SCLC)), medullary thyroid carcinoma (MTC), and mesothelioma. According to some aspects, the solid malignant tumor may be metastatic or unresectable. In some embodiments, the solid malignant tumor may be histologically or cytologically confirmed. The disclosed stable aqueous pharmaceutical composition may be administered subcutaneously, for example, by subcutaneous injection. Subcutaneous injections can be performed at various locations on the subject's body, such as, but not limited to, the upper arm, thigh, abdomen, or lower back.

[0079] In one embodiment, the cancer is lung cancer. In one embodiment, the cancer is non-small cell lung cancer (NSCLC). In one embodiment, the cancer is treatment-naïve locally advanced or metastatic NSCLC. In one embodiment, the cancer has been previously treated with IV amivantamab. In one embodiment, the cancer harbors an EGFR exon 19del mutation. In one embodiment, the cancer harbors an exon 21 L858R mutation. In one embodiment, the cancer harbors an EGFR exon 20ins mutation. In one embodiment, the cancer has experienced disease progression during or after treatment with a third generation EGFR tyrosine kinase inhibitor (TKI).

[0080] Further provided herein is a method of reducing infusion-related reactions in a subject treated with amivantamab, comprising subcutaneously administering to the subject a stable aqueous pharmaceutical formulation disclosed herein, wherein the subject is in need of a cancer treatment as disclosed herein.

[0081] Further provided herein is an article of manufacture containing a stable aqueous pharmaceutical composition of the invention. In one embodiment, the article of manufacture is a single-use glass vial with a stopper containing the stable aqueous pharmaceutical composition to be administered. In some embodiments, the stopper is puncturable by a syringe. In some embodiments, the vial is sealed. In some embodiments, the single-use vial is a 10 mL single-use glass vial with a 20 mm stopper covered with a 20 mm aluminum seal. In some embodiments, the vial size is an ISO format of 2R, 4R, 6R, 8R, 10R, 15R, 20R, 25R, 30R, or 50R with a volume of about 4 mL, 6 mL, 10 mL, 12 mL, 14 mL, 20 mL, 26 mL, 33 mL, 38 mL, or 62 mL, respectively. In one embodiment, the total volume of the stable aqueous pharmaceutical composition (also referred to herein as drug product or DP) ranges from about 5 mL to about 10 mL. In one embodiment, the total volume of the stable aqueous pharmaceutical composition (pharmaceutical product or DP) is in the range of about 0.5 mL to about 20 mL, about 1 mL to about 15 mL, about 5 mL to about 10 mL, or about 6 mL to about 8 mL. In one embodiment, the total volume of the stable aqueous pharmaceutical composition is about 0.5 mL, 0.6 mL, 0.7 mL, 0.8 mL, 0.9 mL, 1 mL, 2 mL, 3 mL, 4 mL, 5 mL, 6 mL, 6.5 mL, 6.6 mL, 6.7 mL, 7 mL, 7.1 mL, 8 mL, 8.5 mL, 8.6 mL, 8.7 mL, 8.75 mL, 8.8 mL, 9 mL, 10 mL, 11 mL, 12 mL, 13 mL, 14 mL, 15 mL, 16 mL, 18 mL, 19 mL, 20 mL, 25 mL, or 30 mL, or any range therebetween.

[0082] Drug stability In some embodiments, the DP stability is determined after storage for a certain period of time. In some embodiments, the DP is stored for about 3 months or more, about 6 months or more, about 12 months or more, about 1.5 months or more, about 2 years or more, about 2.5 years or more, about 3 years or more, about 3.5 years or more, about 4 years or more, about 4.5 years or more, about 5 years or more, about 6 years or more, about 7 years or more, about 8 years or more, about 9 years or more, or about 10 years or more. In some embodiments, the DP is stored for about 12 months or more, about 1.5 years or more, about 2 years or more, about 2.5 years or more, or about 3 years or more. In some embodiments, the DP is stored for about 2 years or more.

[0083] temperature In some embodiments, the DP is stable after storage for a particular period of time at a particular temperature. In some embodiments, the temperature is in the range of about -10-50°C, 0-25°C, 1-20°C, 1-15°C, 2-10°C, or 2-5°C. In some embodiments, the temperature is in the range of about 2-8°C. In some embodiments, the temperature is about -10°C, -9°C, -8°C, -7°C, -6°C, -5°C, -4°C, -3°C, -2°C, -1°C, 0°C, 1°C, 2°C, 3°C, 4°C, 5°C, 6°C, 7°C, 8°C, 9°C, 10°C, 11°C, 12°C, 13°C, 14°C, 15°C, 16°C, 17°C, 18°C, 19°C, 20°C, 21°C, 22°C, 23°C, 24°C, 25°C, 26°C, 27°C, 28°C, 29°C, 30°C, 31°C, 32°C, 33°C, 34°C, 35°C, 36°C, 37°C, 38°C, 39°C, 40°C, 41°C, 42°C, 43°C, 44°C, 45°C, 46°C, 47°C, 48°C, 49°C, or 50°C.

[0084] In some embodiments, the DP is stable after storage for about 12 months or more, or about 2 years or more, at a temperature ranging from about 2° C. to about 8° C. In some embodiments, the DP is stable after storage for about 12 months or more, or about 2 years or more, at a temperature of about 5° C. In some embodiments, the DP is stable after storage for about 12 months or more at a temperature of about 25° C.

[0085] The stability of the aqueous pharmaceutical compositions of the present disclosure, also referred to as drug products (DPs), is determined based on the specific amounts or proportions of the bispecific EGFR-cMet antibodies and other components of the DPs provided herein, such as, but not limited to, buffers, stabilizers, chelating agents, surfactants, and enzymes, as well as an evaluation of a variety of factors, including, but not limited to, solution color, pH, turbidity, number of subvisible particles, percentage of aglycosylated heavy chain (AGHC), percentage of new peaks, percentage of high molecular weight species (HMWS), percentage of low molecular weight species (LMWS), percentage of total acidic peaks, percentage of total basic peaks, protein concentration, percentage of EGFR binding activity, percentage of cMet binding activity, and / or percentage of PS80.

[0086] A stable DP disclosed herein should not be construed as requiring all of the factors listed herein, but rather as requiring at least one, at least two, or at least three or more of the factors. In some embodiments, a stable disclosed DP exhibits the following results for at least one, at least two, at least three, or more of the factors listed in detail herein below. In some embodiments, a stable DP exhibits the following results for all of the factors listed in detail herein below.

[0087] Solution color The color of the DP solution can be monitored and evaluated to verify that the appearance of the solution is consistent with previous batches upon release and over shelf life. The color of the DP solution can reflect stability. In one embodiment, the stability of the DP is defined as having a solution color ranging from colorless to about BY2 or less, about BY4 or less, about B2 or less, about B4 or less, about Y2 or less, or about Y4 or less, as described in European Pharmacopoeia 2.2.2, Degree of Coloration of Liquids European Pharmacopoeia (Ph.Eur.) 10th Edition Monograph No. 20202, July 2019.

[0088] In one embodiment, stability is defined as having a solution color that is colorless to about BY2 or less, about B2 or less, to about Y2 or less after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In a preferred embodiment, stability is defined as having a solution color that is colorless to about BY4 or less, about B4 or less, to about Y4 or less after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In a most preferred embodiment, stability is defined as having a solution color that is colorless to about BY5 or less, about B5 or less, to about Y5 or less after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more.

[0089] pH Measuring the pH of the DP solution can ensure that the pH is consistent with previous DP batches upon release and over the shelf life. In one embodiment, stability of a DP is defined as when its pH is about 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, or 6.4. In one embodiment, the pH of the DP is about 5.7 after about 12 months or more of storage at a temperature of about 5° C., after about 12 months or more of storage at a temperature of about 25° C., and / or after about 2 years or more of storage at a temperature of about 5° C. In one embodiment, stability is defined as having a pH range of about 5.0 to about 6.4 after about 12 months or more of storage at a temperature of about 5° C., after about 12 months or more of storage at a temperature of about 25° C., and / or after about 2 years or more of storage at a temperature of about 5° C. In a preferred embodiment, the stability of the DP is defined as a pH in the range of about 5.2 to about 6.2 after storage for about 12 months or more at a temperature of about 5° C., after storage for about 12 months or more at a temperature of about 25° C., and / or after storage for about 2 years or more at a temperature of about 5° C. In a most preferred embodiment, the stability of the DP is defined as a pH in the range of about 5.4 to about 6.0 after storage for about 12 months or more at a temperature of about 5° C., after storage for about 12 months or more at a temperature of about 25° C., and / or after storage for about 2 years or more at a temperature of about 5° C.

[0090] Turbidity Turbidity allows for the measurement of the presence of particles in the DP solution to ensure consistency with previous DP batches and allows for compendial guidance applicable at the time of release and over the shelf life. Test results are reported in Nephelometric Turbidity Units (NTU). In one embodiment, the stability of a DP is defined as having a turbidity value of about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 Nephelometric Turbidity Units (NTU) after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In one embodiment, the stability of a DP is defined as having a turbidity value of about 18 NTU or less after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In a preferred embodiment, the stability of the DP is defined as having a turbidity value of about 13 NTU or less after storage for about 12 months or more at a temperature of about 5° C., after storage for about 12 months or more at a temperature of about 25° C., and / or after storage for about 2 years or more at a temperature of about 5° C. In a most preferred embodiment, the stability of the DP is defined as having a turbidity value of about 8 NTU or less after storage for about 12 months or more at a temperature of about 5° C., after storage for about 12 months or more at a temperature of about 25° C., and / or after storage for about 2 years or more at a temperature of about 5° C.

[0091] particle analysis The stability of DP is set at a specific threshold of particle contamination based on the average number of particles not visible to the naked eye. The test results are published in the United States Pharmacopeia <788> Particulate Matter, in accordance with European Pharmacopoeia 2.9.19, and Japanese Pharmacopoeia XVII / 6.07 Particulate Contamination: Particles invisible to the naked eye. Therefore, the average number of particles present in the tested DP unit should not exceed 6000 particles per container for particles of 10 μm or greater, and 600 particles per container for particles of 25 μm or greater.

[0092] cSDS conditions Capillary SDS-PAGE (cSDS) is a method that separates denatured proteins based on their molecular weight, similar to gel-based SDS-PAGE. This process allows for quantifying DP purity and monitoring its stability upon release and over shelf life.

[0093] In one embodiment, DP stability is defined based on the results of various cSDS variables (e.g., percent purity, aglycosylated heavy chain (AGHC), or presence of new peaks) after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more under reducing or non-reducing conditions.

[0094] The reduced cSDS results are consistent with stability. In one embodiment, stability is defined as having a percent purity of 88.0% or greater, an AGHC of 11.0% or less, and no new peaks greater than 1.5% compared to a validated stock of amivantamab reference material after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In a preferred embodiment, stability is defined as having a percent purity of about 91.0% or greater, an AGHC of 8.0% or less, and no new peaks greater than 1.0% compared to the reference material after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In the most preferred embodiments, stability is defined as having a percent purity of about 94.0% or greater, an AG HC of about 5.0% or less, and no new peaks greater than 1.0% relative to a reference material after storage at a temperature of about 5° C. for about 12 months or greater, after storage at a temperature of about 25° C. for about 12 months or greater, and / or after storage at a temperature of about 5° C. for about 2 years or greater.

[0095] The results for non-reduced cSDS are consistent with stability. In one embodiment, stability is defined as having a percent purity of about 88.0% or greater and no new peaks greater than 1.5% compared to the reference material after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In a preferred embodiment, stability is defined as having a percent purity of about 90.0% or greater and no new peaks greater than 1.0% compared to the reference material after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In a most preferred embodiment, stability is defined as having a percent purity of about 94.0% or greater and no new peaks greater than 1.0% compared to the reference material after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more.

[0096] In one embodiment, DP stability is defined as having a percent purity of about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or about 100% or equal to 100%, or any range therebetween.

[0097] In one embodiment, DP stability is defined as having an AGHC of about 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, or any range therebetween.

[0098] In one embodiment, DP stability is defined as the absence of more than 0.5%, 0.8%, 0.9%, 1.0%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, or more than 2% new peaks in the cSDS result when compared to an untreated reference material.

[0099] Size-exclusion HPLC (SE-HPLC) results consistent with stability The SE-HPLC procedure makes it possible to assess the purity of the DP and to monitor its stability under non-denaturing conditions upon release and over its shelf life.

[0100] SE-HPLC results are consistent with stability Major Components - In one embodiment, stability is defined as having about 90.0% or greater major components after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In a preferred embodiment, stability is defined as having about 95.0% or greater major components after storage at a temperature of about 12 months or more and about 5° C., after storage at a temperature of about 12 months or more and about 25° C., and / or after storage at a temperature of about 2 years or more and about 5° C. In a most preferred embodiment, stability is defined as having about 97.0% major components after storage at a temperature of about 12 months or more and about 5° C., after storage at a temperature of about 12 months or more and about 25° C., and / or after storage at a temperature of about 2 years or more and about 5° C. High Molecular Weight Species (HMWS) - In one embodiment, stability is defined as having about 10.0% or less HMWS after about 12 months or more of storage at a temperature of about 5° C., after about 12 months or more of storage at a temperature of about 25° C., and / or after about 2 years or more of storage at a temperature of about 5° C. In a preferred embodiment, stability is defined as having about 5.0% or less HMWS after about 12 months or more of storage at a temperature of about 5° C., after about 12 months or more of storage at a temperature of about 25° C., and / or after about 2 years or more of storage at a temperature of about 5° C. In a most preferred embodiment, stability is defined as having about 3.0% or less HMWS after about 12 months or more of storage at a temperature of about 5° C., after about 12 months or more of storage at a temperature of about 25° C., and / or after about 2 years or more of storage at a temperature of about 5° C. Low Molecular Weight Species (LMWS) - In one embodiment, stability is defined as having an LMWS of about 5.0% or less after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In a preferred embodiment, stability is defined as having an LMWS of about 2.0% or less after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In a most preferred embodiment, stability is defined as having an LMWS of about 1.0% or less after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more.

[0101] Capillary isoelectric focusing (cIEF) cIEF separates proteins based on their overall charge or isoelectric point (pI), similar to isoelectric gel electrophoresis (IEF) methods. This procedure allows for monitoring the distribution of charge-based isoforms of a drug product upon release and over shelf life. In one embodiment, DP stability is defined based on the results of various cIEF variables, such as main peak (MP), sum of acidic peaks, or sum of basic peaks, after storage of the DP at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more.

[0102] cIEF results are consistent with stability Main Peak - In one embodiment, DP stability is defined as having a cIEF with an MP of about 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95%, or any range therebetween, after storage of the DP at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In one embodiment, DP stability is defined as having a cIEF with an MP in the range of about 30% to about 90% after storage of the DP at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In one embodiment, stability is defined as having a main peak of 37-87% after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In preferred embodiments, stability is defined as having 47-87% of the main peak after about 12 months or more of storage at a temperature of about 5° C., after about 12 months or more of storage at a temperature of about 25° C., and / or after about 2 years or more of storage at a temperature of about 5° C. In the most preferred embodiments, stability is defined as having 57-87% of the main peak after about 12 months or more of storage at a temperature of about 5° C., after about 12 months or more of storage at a temperature of about 25° C., and / or after about 2 years or more of storage at a temperature of about 5° C.

[0103] Sum of acidic peaks - In one embodiment, DP stability is defined as having a cIEF with a sum of acidic peaks totaling about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, or 70%, or any range therebetween, after storage of the DP at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In one embodiment, stability is defined as having a sum of acidic peaks totaling 10-60% after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In a preferred embodiment, stability is defined as having a sum of acidic peaks totaling 10-50% after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In the most preferred embodiments, stability is defined as having a total of 10-40% total acidic peaks after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more.

[0104] Sum of basic peaks - In one embodiment, DP stability is defined as having a cIEF with a sum of basic peaks totaling about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, or 15%, or any range therebetween, after storage of the DP at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In one embodiment, stability is defined as having a sum of basic peaks totaling about 12.0% or less after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In a preferred embodiment, stability is defined as having a sum of basic peaks totaling about 10.0% or less after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In the most preferred embodiments, stability is defined as having a total of about 8.0% or less of basic peaks after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more.

[0105] Protein Concentration by A280 The protein concentration of the DP allows verification that it matches previous DP batches at the time of shipment and over the shelf life. Protein concentration can be quantified by measuring the UV light absorbance of the pharmaceutical solution at 280 nm (A280).

[0106] The protein concentration results are consistent with the stability of the DP. In one embodiment, DP stability is defined as having a protein concentration of 128-192 mg / mL after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In a preferred embodiment, DP stability is defined as having a protein concentration of 144-176 mg / mL after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In a most preferred embodiment, DP stability is defined as having a protein concentration of 150 mg / mL to 170 mg / mL after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more.

[0107] Drug efficacy In vitro binding of DPs to EGFR and / or c-Met allows for assessment of the level of DP stability, which can be assessed by, but is not limited to, using a homogeneous competitive time-resolved fluorescence resonance energy transfer (TR-FRET) assay.

[0108] The EGFR binding activity results are consistent with stability. In one embodiment, DP stability is defined as having about 40%, 50%, 60%, 70%, 80%, 90%, 100%, 110%, 120%, 130%, 140%, 150%, 160%, or 170%, or any range therebetween, of EGFR binding activity relative to a reference after storage of the DP at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In one embodiment, DP stability is defined as having about 50% to about 150% of EGFR binding activity relative to a reference after storage of the DP at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In a preferred embodiment, DP stability is defined as having an EGFR binding activity in the range of about 60% to about 140% compared to a reference after storage of the DP for about 12 months or more at a temperature of about 25° C. and / or after storage for about 2 years or more at a temperature of about 5° C. In a most preferred embodiment, DP stability is defined as having an EGFR binding activity in the range of about 80% to about 120% compared to a reference after storage of the DP for about 12 months or more at a temperature of about 25° C. and / or after storage for about 2 years or more at a temperature of about 5° C.

[0109] The cMet binding activity results are consistent with stability. In one embodiment, DP stability is defined as having a cMet binding activity of about 40%, 50%, 60%, 70%, 80%, 90%, 100%, 110%, 120%, 130%, or 140%, or any range therebetween, relative to a reference, after storage of the DP at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In one embodiment, DP stability is defined as having a cMet binding activity in the range of about 50% to about 150% relative to a reference, after storage of the DP at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In a preferred embodiment, DP stability is defined as having a cMet binding activity in the range of about 60% to about 140% relative to a reference, after storage of the DP at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In a most preferred embodiment, DP stability is defined as having a cMet binding activity in the range of about 80% to about 120% compared to a reference after storage of the DP at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more.

[0110] Potency rHuPH20 activity In vitro rHuPH20 hyaluronidase enzymatic activity is determined by measuring turbidity when hyaluronic acid (HA), a substrate for rHuPH20, is bound with acidified serum. Hyaluronidase activity determination is based on the formation of a precipitate when hyaluronic acid (HA) is bound with acidified serum. Activity is measured by incubating hyaluronidase with HA for 30 minutes at 37°C in a 96-well plate format, followed by the addition of acidified serum to precipitate undigested HA. The resulting turbidity is measured at 640 nm, and the decrease in turbidity due to enzymatic cleavage of the HA substrate is a measure of hyaluronidase activity.

[0111] The rHuPH20 activity results are consistent with stability. In one embodiment, the stability is about 800 U / mL, 900 U / mL, 1000 U / mL, 1100 U / mL, 1200 U / mL, 1300 U / mL, 1400 U / mL, 1500 U / mL, 1600 U / mL, 1700 U / mL, 1800 U / mL, 1900 U / mL after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. , 2000 U / mL, 2100 U / mL, 2200 U / mL, 2300 U / mL, 2400 U / mL, 2500 U / mL, 2600 U / mL, 2700 U / mL, 2800 U / mL, 2900 U / mL, 3000 U / mL, 3100 U / mL, 3200 U / mL, 3300 U / mL, 3400 U / mL, or 3500 U / mL, or any range therebetween. In one embodiment, stability is defined as having an rHuPH20 activity of 1000 U / mL to 3000 U / mL after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In preferred embodiments, stability is defined as having an rHuPH20 activity of 1500 U / mL to 2500 U / mL after storage for about 12 months or more at a temperature of about 5° C., after storage for about 12 months or more at a temperature of about 25° C., and / or after storage for about 2 years or more at a temperature of about 5° C. In the most preferred embodiments, stability is defined as having an rHuPH20 activity of 1800 U / mL to 2200 U / mL after storage for about 12 months or more at a temperature of about 5° C., after storage for about 12 months or more at a temperature of about 25° C., and / or after storage for about 2 years or more at a temperature of about 5° C.

[0112] Analytical Tests - Surfactants Quantitative determination of polysorbate 80 Polysorbate 80 is quantitatively measured by mixed mode ion exchange / hydrophobic HPLC. In one embodiment, DP stability is defined by a PS80 concentration of about 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.08%, 0.09%, or 0.1%, or any range therebetween, weight to volume percentage, after storage of the DP at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In one embodiment, DP stability is defined as a PS80 concentration of 0.03-0.08% after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In a preferred embodiment, DP stability is defined as a PS80 concentration of 0.04-0.08% after storage for about 12 months or more at a temperature of about 5° C., after storage for about 12 months or more at a temperature of about 25° C., and / or after storage for about 2 years or more at a temperature of about 5° C. In a most preferred embodiment, DP stability is defined as a PS80 concentration of 0.05-0.08% after storage for about 12 months or more at a temperature of about 5° C., after storage for about 12 months or more at a temperature of about 25° C., and / or after storage for about 2 years or more at a temperature of about 5° C.

[0113] viscosity. A potential problem with the use of high concentration protein formulations recognized by those skilled in the art is that the viscosity of the solution typically increases with increasing protein concentration. Viscous antibody solutions are difficult to process (e.g., fill vials and / or syringes) and administer to patients. Highly viscous formulations are difficult to manufacture, draw into syringes, and inject. The use of force in manipulating viscous formulations can result in excessive foaming, leading to denaturation and inactivation of the active biologic. Unless the viscosity can be reduced, highly concentrated antibody formulations may require larger bore needles, high pressure injections, longer injection times, and special equipment or materials to counteract antibody adhesion. These changes increase patient discomfort and the manufacturing costs of therapeutic antibody products.

[0114] As used herein, "viscosity" is the resistance of a fluid to flow and may be measured in centipoise (cP) or millipascal-seconds (mPa-s) at a given shear rate, where 1 cP=1 mPa-s. Viscosity may be measured using a viscometer, such as a Brookfield Engineering Dial Reading Viscometer, models LVT and AR-G2, TA Instruments. Viscosity may also be measured using any other method and in any other units known in the art (e.g. absolute viscosity, kinematic viscosity, or dynamic viscosity), with the understanding that the rate of viscosity reduction obtained by the use of the excipients described by this invention is important. Regardless of the method used to determine viscosity, the rate of viscosity reduction in the control formulation relative to the excipient formulation remains approximately the same at a given shear rate.

[0115] The present invention provides a high-concentration amivantamab composition having a viscosity of about 9 cP to about 11 cP at room temperature. In some embodiments, the high-concentration amivantamab composition has an absolute viscosity of about 22 cP or less, 16 cP or less, 13 cP or less, 11 cP or less, 9 cP or less, 8 cP or less, 7 cP or less, or 6 cP. In some embodiments, the high-concentration amivantamab composition has a viscosity of about 21.9 cP when measured at 4° C. In some embodiments, the high-concentration amivantamab composition has a viscosity of about 16 cP when measured at 10° C. In some embodiments, the high-concentration amivantamab composition has a viscosity of about 13.3 cP when measured at 15° C. In some embodiments, the high-concentration amivantamab composition has a viscosity of about 11 cP when measured at 20° C. In some embodiments, the high-concentration amivantamab composition has a viscosity of about 9.3 cP when measured at 25° C. In some embodiments, the high concentration amivantamab composition has a viscosity of about 7.9 cP when measured at 30° C. In some embodiments, the high concentration amivantamab composition has a viscosity of about 6.7 cP when measured at 35° C. In some embodiments, the high concentration amivantamab composition has a viscosity of about 5.8 cP when measured at 40° C.

[0116] Exemplary embodiments Exemplary embodiments of the disclosed technology are provided herein, which are for illustrative purposes only and are not intended to limit the scope of the disclosure or the claims appended hereto. 1. A stable aqueous pharmaceutical composition comprising a bispecific epidermal growth factor receptor (EGFR) / hepatocyte growth factor receptor (c-Met) antibody and hyaluronidase, wherein the antibody is a. a first heavy chain (HC1) comprising an HC1 variable region 1 (VH1) comprising the amino acid sequence of SEQ ID NO: 13; b. a first light chain (LC1) comprising a light chain variable region 1 (VL1) comprising the amino acid sequence of SEQ ID NO: 14; c. a second heavy chain (HC2) comprising an HC2 variable region 2 (VH2) comprising the amino acid sequence of SEQ ID NO: 15; d. a second light chain (LC2) comprising a light chain variable region 2 (VL2) comprising the amino acid sequence of SEQ ID NO: 16; wherein the composition comprises about 1,050 mg to about 2,240 mg of a bispecific EGFR / c-Met antibody, and about 13,000 U to about 28,000 U of hyaluronidase. 2. The stable composition of embodiment 1, wherein the composition comprises about 1,050 mg of the bispecific EGFR / c-Met antibody. 3. The stable composition of embodiment 1, wherein the composition comprises about 1,400 mg of the bispecific EGFR / c-Met antibody. 4. The stable composition of embodiment 1, wherein the composition comprises about 1,575 mg of the bispecific EGFR / c-Met antibody. 4a. The stable composition of embodiment 1, wherein the composition comprises about 1,600 mg of the bispecific EGFR / c-Met antibody. 5. The stable composition of embodiment 1, wherein the composition comprises about 2,100 mg of the bispecific EGFR / c-Met antibody. 5a. The stable composition of embodiment 1, wherein the composition comprises about 2,240 mg of the bispecific EGFR / c-Met antibody. 6. A stable aqueous pharmaceutical composition comprising: a) a bispecific epidermal growth factor receptor (EGFR) / hepatocyte growth factor receptor (c-Met) antibody at about 144 mg / mL to about 176 mg / mL, the bispecific antibody being a first heavy chain (HC1) comprising an HC1 variable region 1 (VH1); a first light chain (LC1) comprising a light chain variable region 1 (VL1); a second heavy chain (HC2) comprising HC2 variable region 2 (VH2); and a second light chain (LC2) comprising a light chain variable region 2 (VL2); Including, VH1 comprises the heavy chain complementarity determining region 1 (HCDR1), HCDR2, and HCDR3 amino acid sequences of SEQ ID NOs: 1, 2, and 3, respectively; VL1 comprises the light chain complementarity determining region 1 (LCDR1), LCDR2, and LCDR3 amino acid sequences of SEQ ID NOs: 4, 5, and 6, respectively; VH2 comprises the HCDR1, HCDR2, and HCDR3 amino acid sequences of SEQ ID NOs: 7, 8, and 9, respectively; and VL2 comprises the LCDR1, LCDR2, and LCDR3 amino acid sequences of SEQ ID NOs: 10, 11, and 12, respectively; A bispecific antibody; b) about 10 mM to about 50 mM acetic acid and / or a pharma- ceutically acceptable acetate salt; c) about 6.8% (w / v) to about 10.2% (w / v) sucrose; d) about 0.036% (w / v) to about 0.084% (w / v) polysorbate 80 (PS80); e) about 0.8 mg / mL to about 1.2 mg / mL of methionine; f) about 16 μg / mL to about 24 μg / mL ethylenediaminetetraacetic acid (EDTA); g) optionally, about 1,000 U / mL to about 3,000 U / mL of hyaluronidase; h) a pH of about 5.2 to about 6.2; A stable aqueous pharmaceutical composition comprising: 6a. The stable aqueous pharmaceutical composition of embodiment 6, wherein the viscosity of the composition is less than about 11 cP when measured at 20° C. 7. The stable aqueous pharmaceutical composition of embodiment 6, wherein the bispecific EGFR-cMet antibody comprises an HC1 variable region comprising the amino acid sequence of SEQ ID NO: 13 and an LC1 variable region comprising the amino acid sequence of SEQ ID NO: 14. 8. The stable aqueous pharmaceutical composition of embodiment 6 or embodiment 7, wherein the bispecific EGFR-cMet antibody comprises an HC2 variable region comprising the amino acid sequence of SEQ ID NO: 15 and an LC2 variable region comprising the amino acid sequence of SEQ ID NO: 16. 9. The stable aqueous pharmaceutical composition according to any one of embodiments 6 to 8, wherein HC1 comprises the amino acid sequence of SEQ ID NO:17 and LC1 comprises the amino acid sequence of SEQ ID NO:18. 10. The stable aqueous pharmaceutical composition according to any one of embodiments 6 to 9, wherein HC2 comprises the amino acid sequence of SEQ ID NO: 19 and LC2 comprises the amino acid sequence of SEQ ID NO: 20. 11. The stable aqueous pharmaceutical composition of any one of embodiments 6 to 10, wherein the bispecific EGFR-cMet antibody is amivantamab or a biosimilar thereof. 12. The stable aqueous pharmaceutical composition of any one of embodiments 6 to 11, wherein the bispecific EGFR-cMet antibody has a concentration of about 160 mg / mL. 13. The stable aqueous pharmaceutical composition according to any one of embodiments 6 to 12, wherein the acetic acid and / or the pharma-ceutically acceptable acetate salt has a concentration of about 30 mM. 14. The stable aqueous pharmaceutical composition according to any one of embodiments 6 to 13, wherein the acetic acid and / or a pharma-ceutically acceptable acetate comprises glacial acetic acid and / or sodium acetate trihydrate. 15. The stable aqueous pharmaceutical composition of any one of embodiments 6 to 14, comprising about 8.5% (w / v) sucrose. 16. The stable aqueous pharmaceutical composition according to any one of embodiments 6 to 15, comprising about 0.06% (w / v) PS80. 17. The stable aqueous pharmaceutical composition according to any one of embodiments 6 to 16, wherein the methionine comprises L-methionine and has a concentration of about 1 mg / mL. 18. The stable aqueous pharmaceutical composition according to any one of embodiments 6 to 17, wherein EDTA has a concentration of about 20 μg / mL. 19. The stable aqueous pharmaceutical composition according to any one of embodiments 6 to 18, wherein the pH is about 5.7. 20. The stable aqueous pharmaceutical composition of any one of embodiments 6 to 19, wherein the hyaluronidase is a human hyaluronidase, optionally rHuPH20 comprising the amino acid sequence of SEQ ID NO: 21. 21. The stable aqueous pharmaceutical composition of any one of embodiments 6 to 20, wherein the concentration of rHuPH20 is from about 1,000 U / mL to about 3,000 U / mL. 22. The stable aqueous pharmaceutical composition of any one of embodiments 6 to 21, wherein the concentration of rHuPH20 is about 2,000 U / mL. 23. The stable aqueous pharmaceutical composition according to any one of embodiments 6 to 22, wherein the stability is defined based on solution color, pH, turbidity, number of subvisible particles, percentage of aglycosylated heavy chain (AGHC), percentage of new peaks, percentage of high molecular weight species (HMWS), percentage of low molecular weight species (LMWS), percentage of sum of acidic peaks, percentage of sum of basic peaks, protein concentration, percentage of EGFR binding activity, percentage of cMet binding activity, percentage of PS80, optionally percentage of rHuPH20 activity, or any combination thereof. 24. The stable aqueous pharmaceutical composition according to any one of embodiments 6 to 23, wherein the total volume of the composition ranges from about 6 mL to about 9 mL. 25. The stable aqueous pharmaceutical composition of embodiment 24, wherein the total volume of the composition is about 7.1 mL. 26. The stable aqueous pharmaceutical composition of embodiment 24, wherein the total volume of the composition is about 6.6 mL. 27. The stable aqueous pharmaceutical composition of embodiment 24, wherein the total volume of the composition is about 8.75 mL. 28. A stable aqueous pharmaceutical composition comprising about 160 mg / mL of a bispecific EGFR-cMet antibody, about 30 mM acetic acid and / or a pharma- ceutically acceptable acetate salt, about 8.5% sucrose, and about 1 mg / mL L-methionine, together with polysorbate 80 to a final concentration of about 0.06% (w / v) and EDTA to a final concentration of about 20 μg / mL, wherein the stable aqueous pharmaceutical composition has a pH of about 5.7; a bispecific EGFR-cMet antibody comprising a heavy chain 1 (HC1) comprising the amino acid sequence of SEQ ID NO: 17, a HC2 comprising the amino acid sequence of SEQ ID NO: 19, a light chain 1 (LC1) comprising the amino acid sequence of SEQ ID NO: 18, and a LC2 comprising the amino acid sequence of SEQ ID NO: 20; A stable aqueous pharmaceutical composition according to any one of embodiments 1 to 27. 29. A stable aqueous pharmaceutical composition comprising about 160 mg / mL of a bispecific EGFR-cMet antibody, about 30 mM acetic acid and / or a pharma- ceutically acceptable acetate salt, about 8.5% sucrose, about 1 mg / mL L-methionine, together with polysorbate 80 to a final concentration of about 0.06% (w / v) and EDTA to a final concentration of about 20 μg / mL, and rHuPH20 to a final concentration of about 2,000 U / mL, wherein the stable aqueous pharmaceutical composition has a pH of about 5.7; a bispecific EGFR-cMet antibody comprising a heavy chain 1 (HC1) comprising the amino acid sequence of SEQ ID NO: 17, a HC2 comprising the amino acid sequence of SEQ ID NO: 19, a light chain 1 (LC1) comprising the amino acid sequence of SEQ ID NO: 18, and a LC2 comprising the amino acid sequence of SEQ ID NO: 20; A stable aqueous pharmaceutical composition according to any one of embodiments 1 to 27. 30. A method for treating cancer in a subject in need thereof, comprising administering to the subject a pharmaceutical composition according to any one of embodiments 1 to 29. 31. The method of embodiment 30, wherein administration is subcutaneous. 32. The method of embodiment 30-31, wherein the cancer comprises lung cancer, squamous cell carcinoma of the head and neck (SCCHN), hepatocellular carcinoma (HCC), colorectal cancer (CRC), renal cell carcinoma (RCC), medullary thyroid carcinoma (MTC), gastroesophageal carcinoma (GEC), mesothelioma, breast cancer (BC), or ovarian cancer (OC). 33. The method of embodiment 30-31, wherein the cancer comprises non-small cell lung cancer (NSCLC). 34. A method for preparing a stable aqueous pharmaceutical composition of a bispecific antibody targeting EGFR and cMet, the bispecific antibody targeting EGFR and cMet comprising a first heavy chain (HC1) comprising HC1 variable region 1 (VH1), a first light chain (LC1) comprising light chain variable region 1 (VL1), a second heavy chain (HC2) comprising HC2 variable region 2 (VH2), and a second light chain (LC2) comprising light chain variable region 2 (VL2), wherein VH1 is selected from the group consisting of SEQ ID NOs: 1, 2, and 3, respectively. wherein VL1 comprises a heavy chain complementarity determining region 1 (HCDR1), HCDR2, and HCDR3 comprising the amino acid sequences of SEQ ID NOs: 4, 5, and 6, respectively; VH2 comprises the HCDR1, HCDR2, and HCDR3 amino acid sequences of SEQ ID NOs: 7, 8, and 9, respectively; and VL2 comprises the LCDR1, LCDR2, and LCDR3 amino acid sequences of SEQ ID NOs: 10, 11, and 12, respectively; and the method comprises 1. A method comprising combining a composition comprising about 160 mg / mL of a bispecific antibody, about 30 mM acetic acid and / or a pharma- ceutically acceptable acetate salt, about 8.5% sucrose, and about 1 mg / mL of L-methionine with polysorbate 80 to a final concentration of about 0.06% (w / v) and EDTA to a final concentration of about 20 μg / mL, optionally rHuPH20 to a final concentration of about 2,000 U / mL, wherein the stable aqueous pharmaceutical composition has a pH of about 5.7. 35. The method of embodiment 34, wherein the bispecific EGFR-cMet antibody comprises an HC1 variable region comprising the amino acid sequence of SEQ ID NO: 13 and an LC1 variable region comprising the amino acid sequence of SEQ ID NO: 14. 36. The method of embodiment 34-35, wherein the bispecific EGFR-cMet antibody comprises an HC2 variable region comprising the amino acid sequence of SEQ ID NO:15 and an LC2 variable region comprising the amino acid sequence of SEQ ID NO:16. 37. The method according to any one of embodiments 34 to 36, wherein the antibody comprises heavy chain 1 (HC1) comprising the amino acid sequence of SEQ ID NO: 17 and light chain 1 (LC1) comprising the amino acid sequence of SEQ ID NO: 18. 38. The method of any one of embodiments 34 to 37, wherein the antibody comprises an HC2 comprising the amino acid sequence of SEQ ID NO: 19 and an LC2 comprising the amino acid sequence of SEQ ID NO: 20. 39. The method of any one of embodiments 34 to 38, wherein the antibody is amivantamab or a biosimilar thereof. 40. A kit comprising the stable aqueous pharmaceutical composition according to any one of embodiments 1 to 29 and instructions for use thereof. 41. An article of manufacture comprising a container holding a stable aqueous pharmaceutical composition according to any one of embodiments 1-29. 42. The article of manufacture of embodiment 41, wherein the container is a vial having a stopper pierceable by a syringe. 43. The article of manufacture of embodiment 42, wherein the vial is a single-use vial. 44. A pharmaceutical composition according to any one of embodiments 1 to 29, for use in the treatment of cancer. 45. The pharmaceutical composition of embodiment 44, wherein the cancer comprises lung cancer. 46. ​​The pharmaceutical composition of embodiment 44, wherein the cancer comprises non-small cell lung cancer (NSCLC). 47. A pharmaceutical composition according to any one of embodiments 1 to 29 for use in the preparation of a medicament for treating cancer. 48. Use of a pharmaceutical composition for treating cancer in a subject in need of treatment by administering the pharmaceutical composition according to any one of embodiments 1 to 29. 49. Use of the pharmaceutical composition according to embodiment 48, wherein administration is subcutaneous. 50. Use of the pharmaceutical composition according to embodiment 49 to reduce infusion-related reactions in a subject treated with amivantamab. EXAMPLES

[0117] The following examples are provided to further illustrate some of the embodiments disclosed herein and are intended to be illustrative and not limiting of the embodiments of the present disclosure.

[0118] Description of analytical tests used herein Analytical Tests - General Characteristics Evaluation Solution color Solution color is monitored for drug products (DP) to assess appearance and ensure consistency with previous batches upon release and over shelf life. Solution color can also be an indicator of product stability. To determine solution color, test samples are visually compared to a set of defined reference solutions.

[0119] A defined amount of the liquid contents is transferred into a pre-scored ampoule of the same dimensions as the reference solution. The contents of the ampoule are then visually compared to the European Pharmacopoeia color reference solutions. The degree of color is judged in diffuse daylight observed against a white background.

[0120] Solution material color and method Materials and methods are described in European Pharmacopoeia 2.2.2, Degree of Coloration of Liquids European Pharmacopoeia (Ph.Eur.) 10th Edition Monograph No. 20202, July 2019. Briefly, test articles are compared to a set of B (brown), BY (brownish yellow), and Y (yellow) color reference solutions.

[0121] Solution color results are consistent with stability In one embodiment, the stability of the DP is defined as having a solution color of colorless to about BY2 or less, about B2 or less, about Y2 or less after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In a preferred embodiment, the stability of the DP is defined as having a solution color of colorless to about BY4 or less, about B4 or less, about Y4 or less after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In a most preferred embodiment, the stability of the DP is defined as having a solution color of colorless to about BY5 or less, about B5 or less, about Y5 or less after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more.

[0122] pH pHMaterials and Methods The pH of the test articles is measured using a daily calibrated electronic pH meter with a standardized pH electrode. All calibration solutions, reference buffers, and test articles are equilibrated to 25°C prior to testing and maintained at 25°C during testing.

[0123] pH results are consistent with stability In one embodiment, the stability of the DP is defined as having a pH range of 5.0 to 6.4 after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In a preferred embodiment, the stability of the DP is defined as having a pH range of 5.2 to 6.2 after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In a most preferred embodiment, the stability of the DP is defined as having a pH range of 5.4 to 6.0 after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more.

[0124] Turbidity Turbidity Materials and Methods Materials and methods are based on the European Pharmacopoeia 2.2.1, Clarity and Degree of Opalescence of Liquids.

[0125] The turbidity results are consistent with stability. Test results are reported in Nephelometric Turbidity Units (NTU). In one embodiment, the stability of the DP is defined as having a turbidity value of about 18 NTU or less after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In a preferred embodiment, the stability of the DP is defined as having a turbidity value of about 13 NTU or less after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In a most preferred embodiment, the stability of the DP is defined as having a turbidity value of about 8 NTU or less after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more.

[0126] Analytical Testing - Particulate Matter Particulate matter (not visible to the naked eye) Materials and Methods - All materials and methods conform to the United States Pharmacopoeia <788> Compendial liquid particle counters equipped with compendial volumetric sampler devices are used. Test articles are equilibrated to room temperature for at least 60 minutes but not more than 10 hours prior to testing. Test article vials are measured using a method consistent with the United States Pharmacopeia (USP) Particulate Matter (EPM) standard. <788> Pool in a manner consistent with Particulate Matter. <788> Remove four portions of appropriate volume of each of the pooled test articles as instructed by Particulate Matter and count the number of particles 10 μm and 25 μm or larger per portion. Disregard the result obtained for the first portion and use the results of the remaining three to calculate the average particle count for the preparation tested.

[0127] Particle Analysis (not visible to the naked eye) Compendial Results - Test results are in accordance with the United States Pharmacopoeia <788> Particulate Matter, in accordance with European Pharmacopoeia 2.9.19, and Japanese Pharmacopoeia XVII / 6.07 Particulate Contamination: Particles invisible to the naked eye. Therefore, the average number of particles present in the tested unit should not exceed 6000 particles per container for particles of 10 μm or greater, and 600 particles per container for particles of 25 μm or greater.

[0128] Analytical Testing - Purity Capillary electrophoresis sodium dodecyl sulfate (cSDS)-reduced form Reduced cSDS Materials and Methods - The analysis uses a commercially available capillary electrophoresis system with a bare fused silica capillary, 50 μm ID x 30.2 cm length, in a temperature-controlled cartridge. The capillary is equipped with a detection window transparent to UV light. The capillary is electrokinetically rinsed before each injection. A sieving matrix consisting of an entangled polymer solution is loaded into the capillary prior to each sample analysis. The method utilizes SDS-MW gel running buffer and protein molecular weight authentication standards ranging from approximately 10 to 148 kDa. The UV absorption spectrophotometer detector of the instrument is set to a wavelength of 220 nm and the capillary temperature is set to 25°C. For reduced sample processing conditions, the test article (in duplicate) is mixed with SDS and 2-mercaptoethanol and then heated for a specified time and temperature to completely denature and reduce the protein. The reduced sample is electrokinetically injected by applying a voltage of 5 kV to the capillary for approximately 20 seconds, and then analyzed by applying a larger electric field for approximately 35 minutes. Detection is achieved by absorbance in the far UV region of the spectrum at 220 nm. Percent of total signal data is collected for the light chain, heavy chain, and aglycosylated heavy chain (AG HC).

[0129] The reduced cSDS results are consistent with stability. In one embodiment, the stability of the DP is defined as having a percent purity of 88.0% or greater, an AG HC of 11.0% or less, and no new peaks greater than 1.5% compared to a validated stock of amivantamab reference material after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In a preferred embodiment, the stability of the DP is defined as having a percent purity of about 91.0% or greater, an AG HC of 8.0% or less, and no new peaks greater than 1.0% compared to the reference material after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In a most preferred embodiment, the stability of the DP is defined as having a percent purity of about 94.0% or greater, an AG HC of about 5.0% or less, and no new peaks greater than 1.0% relative to a reference material after storage at a temperature of about 5° C. for about 12 months or greater, after storage at a temperature of about 25° C. for about 12 months or greater, and / or after storage at a temperature of about 5° C. for about 2 years or greater.

[0130] Capillary electrophoresis sodium dodecyl sulfate (cSDS) - non-reducing Non-reduced cSDS Materials and Methods - The analysis uses a commercially available capillary electrophoresis system with a bare fused silica capillary, 50 μm inner diameter x 30.2 cm length, in a temperature-controlled cartridge. The capillary is equipped with a detection window transparent to UV light. The capillary is electrokinetically rinsed before each injection. A sieving matrix consisting of an entangled polymer solution is loaded into the capillary prior to each sample analysis. The method utilizes SDS-MW gel running buffer, protein molecular weight authentication standards spanning the range of approximately 10-148 kDa, and a validated amivantamab reference material sample. The UV absorption spectrophotometer detector of the instrument is set to a wavelength of 220 nm and the capillary temperature is set to 25 °C. For non-reduced sample processing conditions, the test article (in duplicate) is mixed with SDS and an alkylating reagent (N-ethylmaleimide, to prevent shuffling or reformation of disulfide bonds). It is then heated for a specified time and temperature to completely denature the protein and minimize the formation of fragments and artifact bands. The non-reduced sample is electrokinetically injected by applying a voltage of 5 kV to the capillary for about 20 seconds, and then analyzed by applying a larger electric field for about 35 minutes. Detection is achieved by absorbance in the far-UV region of the spectrum at 220 nm. The percent of total signal data is collected. The data is also analyzed for the presence of new peaks compared to amivantamab reference material. The percent purity is defined as percent heavy chain + percent light chain.

[0131] The results for non-reduced cSDS are consistent with stability. In one embodiment, the stability of the DP is defined as having a percent purity of about 88.0% or greater and no new peaks of more than 1.5% compared to the reference material after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In a preferred embodiment, the stability of the DP is defined as having a percent purity of about 90.0% or greater and no new peaks of more than 1.0% compared to the reference material after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In a most preferred embodiment, the stability of the DP is defined as having a percent purity of about 94.0% or greater and no new peaks of more than 1.0% compared to the reference material after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more.

[0132] Size Exclusion High Performance Liquid Chromatography (SE-HPLC) SE-HPLC Materials and Methods - Reference materials and test articles are diluted to the target protein concentration. A volume of 20 μL of analyte is injected into a 7.8 mm x 30 cm size exclusion column with a silica base of 5 μm particle size, with a fractionation range of 10-500 kDa. Aqueous phosphate buffer is used as the mobile phase at a flow rate of 0.7 mL / min, and the absorbance of the eluate is continuously monitored at 280 nm. Monomers (major components or main peaks), aggregates (high molecular weight species, or HMWS), and fragments (low molecular weight species, or LMWS) are separated on the column and elute at different retention times. The amounts of these species are measured by monitoring the peak absorbance at 280 nm.

[0133] SE-HPLC results are consistent with stability Major Components - In one embodiment, the stability of the DP is defined as having about 90.0% or greater major components after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In a preferred embodiment, the stability of the DP is defined as having about 95.0% or greater major components after storage at a temperature of about 12 months or more and about 5° C., after storage at a temperature of about 12 months or more and about 25° C., and / or after storage at a temperature of about 2 years or more and about 5° C. In a most preferred embodiment, the stability of the DP is defined as having about 97.0% major components after storage at a temperature of about 12 months or more and about 5° C., after storage at a temperature of about 12 months or more and about 25° C., and / or after storage at a temperature of about 2 years or more and about 5° C. High Molecular Weight Species (HMWS) - In one embodiment, the stability of the DP is defined as having an HMWS of about 10.0% or less after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In a preferred embodiment, the stability of the DP is defined as having an HMWS of about 5.0% or less after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In a most preferred embodiment, the stability of the DP is defined as having an HMWS of about 3.0% or less after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. Low Molecular Weight Species (LMWS) - In one embodiment, the stability of the DP is defined as having an LMWS of about 5.0% or less after storage for about 12 months or more at a temperature of about 5° C., after storage for about 12 months or more at a temperature of about 25° C., and / or after storage for about 2 years or more at a temperature of about 5° C. In a preferred embodiment, the stability of the DP is defined as having an LMWS of about 2.0% or less after storage for about 12 months or more at a temperature of about 5° C., after storage for about 12 months or more at a temperature of about 25° C., and / or after storage for about 2 years or more at a temperature of about 5° C.In the most preferred embodiments, the stability of the DP is defined as having an LMWS of about 1.0% or less after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more.

[0134] Capillary isoelectric focusing (cIEF) cIEF Materials and Methods - The analytical procedure is performed on a commercial imaging cIEF analyzer equipped with an autosampler. The analysis uses 100 μm inner wall coated silica capillaries with an outer wall polyimide coating. In addition, an analyte solution of dilute phosphoric acid and methylcellulose, a catholyte solution of sodium hydroxide and methylcellulose, and ampholytes of predetermined type and amount are used. The specimens are treated with carboxypeptidase B (CPB) to remove the C-terminal lysine and eliminate ambiguity introduced by the presence of multiple C-terminal variants for each charged species. The instrument's autosampler is set at 4° C. for both pre-run and run. The pre-run voltage and time are 1500 V and 1 min, respectively. The run voltage and time are 3000 V and 7 min, respectively.

[0135] cIEF results are consistent with stability Main Peak - In one embodiment, the stability of the DP is defined as having 37-87% of the main peak after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In a preferred embodiment, the stability of the DP is defined as having 47-87% of the main peak after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In a most preferred embodiment, the stability of the DP is defined as having 57-87% of the main peak after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more.

[0136] Sum of Acidic Peaks - In one embodiment, the stability of the DP is defined as having a sum of acidic peaks of 10-60% total after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In a preferred embodiment, the stability of the DP is defined as having a sum of acidic peaks of 10-50% total after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In a most preferred embodiment, the stability of the DP is defined as having a sum of acidic peaks of 10-40% total after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more.

[0137] Sum of Basic Peaks - In one embodiment, the stability of the DP is defined as having a sum of basic peaks of about 12.0% or less after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In a preferred embodiment, the stability of the DP is defined as having a sum of basic peaks of about 10.0% or less after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In a most preferred embodiment, the stability of the DP is defined as having a sum of basic peaks of about 8.0% or less after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more.

[0138] Analytical Test - Quantity Protein Concentration by A280 The protein concentration of the preparation is determined by quantification of absorbance at 280 nm (A280).

[0139] Protein Concentration by A280 Materials and Methods Protein concentration is measured using a qualified and calibrated double beam UV-Vis spectrophotometer. Test articles are diluted 1:125 using 0.9% (w / v) NaCl. Samples are measured using quartz semi-microcuvette (1.4 mL) with 1 cm path length and black or matte surface. The spectrophotometer is set at 280 nm wavelength, 1 nm slit width, and 1 second response. 0.9% (w / v) NaCl is used as a blank control. Protein concentration (mg / mL) is calculated by multiplying the absorbance of the test article by the dilution factor, the extinction coefficient of the antibody, and the path length of the instrument (e.g., but not limited to, 1.40 extinction coefficient for amivantamab (mg / mL)). -1 cm -1 It is calculated by dividing the measurement time by the product of the measurement time and the path length of the instrument (1 cm).

[0140] Protein concentration results are consistent with DP stability In one embodiment, the stability of the DP is defined as having a protein concentration of 128-192 mg / mL after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In a preferred embodiment, the stability of the DP is defined as having a protein concentration of 144-176 mg / mL after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In a most preferred embodiment, the stability of the DP is defined as having a protein concentration of 150 mg / mL-170 mg / mL after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more.

[0141] Analytical Testing - Potency Potency (Epidermal Growth Factor Receptor (EGFR) binding) In vitro binding of pharmaceuticals to EGFR is demonstrated using a homogeneous competitive time-resolved fluorescence resonance energy transfer (TR-FRET) assay format. In this procedure, various concentrations of unlabeled bispecific EGFR-cMet antibody samples compete with donor fluorophore (Europium (Eu) chelate)-labeled bispecific EGFR-cMet antibody for binding to acceptor fluorophore (Cy5)-labeled EGFR antigen. Excitation of the donor fluorophore results in the transfer of energy to the bound acceptor fluorophore (FRET process). The resulting FRET is detected by the emission of light at 665 nm using a microplate reader capable of measuring time-resolved fluorescence. Sample dose-response curves are compared to RM.

[0142] EGFR Binding Materials and Methods. Authenticated, commercially available EGFR, recombinant human EGFR / ErbB1 / HER1 with a C-terminal His tag, is reacted with an authenticated, commercially available Cy5 Mono NHS Ester to produce Cy5-labeled EGFR. A validated bispecific EGFR-cMet antibody is reacted with an authenticated, commercially available Europium (Eu) chelate to produce Eu-labeled bispecific EGFR-cMet antibody. Serial dilutions of bispecific EGFR-cMet antibody reference material (RM), assay control, and test article are tested in parallel on the same assay plate. After Eu-labeled bispecific EGFR-cMet antibody is added to each RM, assay control, and test article, the assay plate is gently shaken. Cy5-EGFR is then similarly added, the assay plate is again gently shaken, and incubated in the dark for 4±1 hours. Fluorescence is then measured spectrophotometrically at 665 nm, plotted against antibody concentration, and analyzed by a four-parameter logistic model. The antibody concentration required to obtain half the maximum fluorescence response (EC50) is determined for the RM, assay controls and samples. The potency of the assay controls and samples is calculated based on the ratio of the sample (or control) to the RM EC50 values ​​and reported as a percentage of activity relative to the RM.

[0143] The results of EGFR binding activity are consistent with stability. In one embodiment, the stability of a DP is defined as 50% to 150% of the binding activity compared to the reference material after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In a preferred embodiment, the stability of a DP is defined as 60% to 140% of the binding activity compared to the reference material after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In a most preferred embodiment, the stability of a DP is defined as a binding activity in the range of about 80% to 120% compared to the reference material after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more.

[0144] Potency (cMet binding) In vitro binding of bispecific EGFR-cMet antibodies to c-MET is demonstrated using a homogeneous competitive time-resolved fluorescence resonance energy transfer (TR-FRET) assay format. In this procedure, various concentrations of unlabeled bispecific EGFR-cMet antibody samples compete with donor fluorophore (Europium (Eu) chelate)-labeled bispecific EGFR-cMet antibodies for binding to acceptor fluorophore (Cy5)-labeled c-MET antigen. Excitation of the donor fluorophore results in the transfer of energy to the bound acceptor fluorophore (FRET process). The resulting FRET is detected by the emission of light at 665 nm using a microplate reader capable of measuring time-resolved fluorescence. Sample dose-response curves are compared to a reference material (RM).

[0145] c-MET Binding Materials and Methods. Authenticated, commercially available cMet, a recombinant cMet / HGFR with a c-terminal His tag, is reacted with an authenticated, commercially available Cy5 Mono NHS Ester to generate Cy5-labeled c-MET. A validated bispecific EGFR-cMet antibody is reacted with an authenticated, commercially available europium (Eu) chelate to generate Eu-labeled bispecific EGFR-cMet antibody. Serial dilutions of bispecific EGFR-cMet antibody RM, assay control, and test article are tested in parallel on the same assay plate. After Eu-labeled bispecific EGFR-cMet antibody is added to each RM, assay control, and test article, the assay plate is gently shaken. Cy5-c-MET is then similarly added, the assay plate is again gently shaken, and incubated in the dark for 4±1 hours. Fluorescence is then measured spectrophotometrically at 665 nm, plotted against antibody concentration, and analyzed by a four-parameter logistic model. The antibody concentration required to obtain half the maximum fluorescence response (EC50) is determined for the RM, assay controls and samples. The potency of the assay controls and samples is calculated based on the ratio of the sample (or control) to the RM EC50 values ​​and reported as a percentage of activity relative to the RM.

[0146] The results of cMet binding activity are consistent with stability. In one embodiment, the stability of a DP is defined as a binding activity in the range of about 50% to about 150% compared to the reference material after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In a preferred embodiment, the stability of a DP is defined as a binding activity in the range of about 60% to about 140% compared to the reference material after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In a most preferred embodiment, the stability of a DP is defined as a binding activity in the range of about 80% to about 120% compared to the reference material after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more.

[0147] Potency rHuPH20 activity In vitro rHuPH20 hyaluronidase enzymatic activity is determined by measuring turbidity when hyaluronic acid (HA), a substrate for rHuPH20, is bound with acidified serum. Hyaluronidase activity determination is based on the formation of a precipitate when hyaluronic acid (HA) is bound with acidified serum. Activity is measured by incubating hyaluronidase with HA for 30 minutes at 37°C in a 96-well plate format, followed by the addition of acidified serum to precipitate undigested HA. The resulting turbidity is measured at 640 nm, and the decrease in turbidity due to enzymatic cleavage of the HA substrate is a measure of hyaluronidase activity.

[0148] rHuPH20 Activity Materials and Methods The assay method is based on the United States Pharmacopeia Monograph USP29-NF24 Hyaluronidase for Injection. Standard reagents include 500 mM acetate buffer (pH 3.1), 100 mM acetate buffer (pH 3.1), sterile water for irrigation (SWFI), human serum albumin (HSA) 25% (NDC#68209-643-02), horse serum, 50 mg / mL sodium hyaluronate, and rHuPH20. Assay specific reagents are listed in Table 1 below. Prepare enzyme diluent, horse serum working solution (2.8%), and 0.7 mg / mL HA substrate on the day of the assay. Store HA substrate tubes at 2-8 °C until ready for use.

[0149] [Table 1] 1) 2-(N-morpholino)ethanesulfonic acid hydrate, 4-morpholineethanesulfonic acid

[0150] Assay and sample preparation Prior to starting the assay, an empty 96-well reaction plate is placed in an Eppendorf Thermo-mixer set to 15° C. and allowed to equilibrate for a minimum of 30 minutes, and a heat block is placed in a 37° C. incubator and allowed to equilibrate at the appropriate temperature for at least 2 hours before starting the enzymatic reaction.

[0151] Test samples with an expected rHuPH20 activity of 2,000 U / mL are diluted to 9.3 U / mL in enzyme diluent. Aliquots of rHuPH20 controls and diluted test samples are loaded into a 96-well transfer plate. In a separate dilution plate, designated volumes of enzyme diluent are aliquoted into designated wells. WRS is aliquoted in triplicate into designated wells and then serially diluted in designated wells containing enzyme diluent. Data from these wells are used to generate a 6-point calibration curve. Fixed volumes of rHuPH20 controls and diluted samples are transferred in duplicate from the transfer plate to designated wells in the dilution plate.

[0152] Enzyme reaction Remove HA Substrate Solution from 2-8°C storage and mix gently by inversion 3-4 times. While maintaining the equilibrated 96-well reaction plate in the Thermo-mixer at 15°C, aliquot a fixed volume of HA Substrate Solution into the corresponding well locations of the dilution plate. The standards, controls and samples from the dilution plate are then transferred to the corresponding well locations of the dilution plate. The reaction plate is then mixed at 900 rpm for 10 seconds using a 15°C Thermo-mixer. Remove the reaction plate from the Thermo-mixer, place the plate lid on the reaction plate and immediately transfer to a pre-incubated heat block in a 37°C incubator. Incubate the reaction plate at 37°C for 30±5 minutes.

[0153] Cessation of reactions and development Once the 37°C incubation is complete, remove the covered reaction plate from the incubator and immediately push the covered plate into freshly dispensed ice in an ice bucket and start a timer for 2 minutes. Leave the plate untouched for the 2 minute incubation. After 2 minutes, wipe the bottom of the plate (e.g., with a Kimwipe) to remove any condensation or water and transfer the reaction plate to a 15°C Thermo-mixer. Place the Thermo-mixer lid on top and incubate at 15°C for 10 minutes.

[0154] After 10 minutes, the Thermo-mixer lid is immediately removed and a fixed volume of Horse Serum Working Solution is aliquoted into each well. The plate is then covered with the Thermo-mixer lid and a timer is started for 20±5 minutes at 15°C. After 20±5 minutes of incubation, the reaction plate is transferred from the Thermo-mixer to a 96-well plate reader at 15°C. The optical density of the samples is then measured at 640 nm.

[0155] 4. Data Analysis The known rHuPH20 activity (U / mL) values ​​of the serially diluted WRS samples are plotted against their corresponding measured optical density (OD) values ​​to obtain a curve-fit equation. The rHuPH20 activity of each sample dilution in the wells of the 96-well plate is calculated by using the individual OD values ​​and the curve-fit equation obtained from the reference material curve.

[0156] The rHuPH20 activity results are consistent with the stability of the DP. In one embodiment, the stability of the DP is defined as having an rHuPH20 activity of 1000 U / mL to 3000 U / mL after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In a preferred embodiment, the stability of the DP is defined as having an rHuPH20 activity of 1500 U / mL to 2500 U / mL after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In a most preferred embodiment, the stability of the DP is defined as having an rHuPH20 activity of 1800 U / mL to 2200 U / mL after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more.

[0157] Analytical Tests - Surfactants Quantitative determination of polysorbate 80 Polysorbate 80 is quantitatively measured by mixed mode ion exchange / hydrophobic HPLC.

[0158] PS80 Materials and Methods. The analysis is performed on a gradient HPLC equipped with a 2.1 x 20 mm online column containing 30 μm water-wettable mixed-mode polymeric spherical sorbent particles, an ELSD, and a temperature-controlled column section at 30°C. The flow rate is set at 1 mL / min and the ELSD evaporator temperature is set at 50°C. Mobile phase A is 2% v / v formic acid in water and mobile phase B is 2% v / v formic acid in isopropyl alcohol. Neat polysorbate 80 is used to make calibration and check standards. Test article samples are injected undiluted.

[0159] The polysorbate 80 results are consistent with the stability of the DP. In one embodiment, the stability of the DP is defined as a PS80 concentration of 0.03-0.08% after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In a preferred embodiment, the stability of the DP is defined as a PS80 concentration of 0.04-0.08% after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more. In a most preferred embodiment, the stability of the DP is defined as a PS80 concentration of 0.05-0.08% after storage at a temperature of about 5° C. for about 12 months or more, after storage at a temperature of about 25° C. for about 12 months or more, and / or after storage at a temperature of about 5° C. for about 2 years or more.

[0160] Analytical Testing - Routine Characterization Peptide Map The purpose of this test is to measure the level of post-translational modifications such as oxidation, deamidation, and isomerization that may be present in the antibody structure. The test article is enzymatically digested to obtain peptide segments. These peptides are then evaluated by ultra-performance liquid chromatography mass spectrometry (UPLC-MS). Each peptide sequence analyzed is identified relative to its known location within the overall antibody structure. Post-translational modifications are measured by comparing the measured mass of the identified peptide sequence with its expected mass.

[0161] Peptide Mapping Materials and Methods. Samples are denatured with 6M guanidine, 50mM Tris pH 8.0, 5mM EDTA and filtered using a 30kDa centrifugal filter device (flow-through discarded). Denatured samples are reduced with 1M dithiothreitol (DTT) followed by alkylation with 1M sodium iodoacetate and further treatment with DTT to quench the reaction. The reaction mixture is exchanged into digestion buffer (50mM Tris pH 7.0 with 1mM CaCl2) through a Sephadex G-25 column with separate columns used for blanks, reference material, and test articles. An aliquot of 1mg / mL trypsin stock solution is added to the sample in digestion buffer to give a trypsin concentration of 20μL / mL. The solution is incubated at 37°C for 2 hours ± 30 minutes. The trypsinized solution is cooled to room temperature and the enzyme is inactivated with trifluoroacetic acid. The processed samples are evaluated by ultra-performance liquid chromatography-mass spectrometry (UPLC-MS) equipped with a Waters Acquity BEH (Ethylene Bridged Hybrid) C18, 2.1 x 100 mm, 1.7 μm, 130 Å column and an attached autosampler. Mobile phase A is 0.1% formic acid in water and mobile phase B is 0.1% FA in acetonitrile (mobile phase B). The autosampler is set to 2-8 °C, the column is set to 40 °C, and the flow rate is set to 500 μL / min. Eluted peptides were subjected to electrospray ionization and detected using a calibrated online mass spectrometry.

[0162] Example 1: Formulation Screening Study Two formulation screening studies were conducted to evaluate the stability trends of high concentration (175 mg / mL) amivantamab formulated at various pH values, formulation buffer types, and buffer concentrations.

[0163] Research 1 The first study evaluated test formulations consisting of a range of pH values ​​and corresponding buffer types with all other formulation components held at fixed values ​​(Table 2).

[0164] [Table 2]

[0165] The test formulations were held under normal (5° C.) and stress (40° C.) stability conditions for three weeks. The test formulations were then assayed by SEC to assess the percent aggregates, monomers, and fragments observed for each test formulation.

[0166] As shown in Table 3 below, increasing pH values ​​generally correlated with increasing % aggregates under both normal (5° C.) and stressed (40° C.) stability conditions. As expected, aggregation values ​​were greater under stressed conditions. Overall, formulations 3 and 4 performed less well than formulations 1 and 2. Therefore, the high pH formulations and phosphate buffer were not considered for further development.

[0167] Under normal conditions, both formulations 1 and 2 exhibited similar results consistent with stable formulations with formulation 1 exhibiting slightly less aggregation than formulation 2. However, under stressed conditions, formulation 2 exhibited the least aggregation despite an increase in fragmentation. Based on the seemingly comparable tradeoff between aggregation and fragmentation, both acetate and histidine buffers at low pH values ​​were considered for further evaluation.

[0168] [Table 3]

[0169] Research 2 In the second study, buffer concentrations were evaluated. A histidine buffer at pH 5.6 was selected as a representative low pH formulation. All other formulation components were held at fixed values ​​(Table 4).

[0170] [Table 4]

[0171] The test formulations were held under stressed (40° C.) stability conditions for 3 weeks. The test formulations were then assayed by SEC to assess the percent of aggregates, monomers, and fragments observed for each test formulation.

[0172] As shown below in Table 5, increasing buffer concentrations correlated with decreasing % aggregates. Therefore, formulations with a range of buffer concentrations will be considered for further evaluation.

[0173] [Table 5]

[0174] Example 2: High concentration stability test Stability studies were performed to evaluate three high protein concentration formulations with various buffer concentrations and species (see Table 6 below). Test formulations were held at recommended (5° C.), accelerated (25° C.), and stressed (40° C.) conditions for up to six months.

[0175] [Table 6]

[0176] Three test formulations were prepared from a 50 mg / mL stock formulation of amivantamab. The stock formulation was concentrated by tangential flow filtration (TFF) and buffer exchanged by ultrafiltration / diafiltration (UF / DF). The test formulation was aliquoted into 30R vials with a fill volume of 15.6 mL. The vials were stoppered, capped, and crimp sealed. The vials were placed in a stable location under recommended (5° C.), accelerated (25° C.), and stress (40° C.) conditions. Samples are taken and assayed at the designated time points.

[0177] Test results Stability results of the test formulations held under recommended, accelerated, and stressed conditions are listed below in Tables 7, 8, and 9. At the start (T=0) and throughout the study, it was noted that the measured pH value of formulation 3 did not match its target value. Samples of formulation 3 were assayed for acetate concentration, which was reported to be 30 mM. The shift in pH and acetate concentration was the result of the Gibbs-Donnan effect, the magnitude of which was influenced by the high protein concentration, the pI of the protein, and the pH of the diafiltration buffer used. Thus, the actual composition of formulation 3 was 160 mg / mL amivantamab in 30 mM acetate, 8.5% sucrose, 1 mg / mL methionine, 20 μg / mL EDTA, 0.06% PS-80, pH 5.7.

[0178] Stability at 5℃ All three formulations showed little change in attributes over time: any changes observed were small, consistent with protein degradation over time at 5°C, and of similar magnitude across all formulations.

[0179] Stability at 25°C All three formulations showed little to minimal change in attribute values ​​over time consistent with protein degradation over time at 25° C., with similar magnitude of change observed across all formulations except for the cIEF attribute values. All formulations showed a steady increase over time in the total % acidic peaks and a corresponding decrease in the main peak %. The magnitude of change over time was similar for formulations 2 and 3, while the magnitude was significantly greater for formulation 1.

[0180] Stability at 40°C All three formulations showed significant changes in attribute values ​​over time, consistent with protein degradation over time for up to 6 months at 40° C. However, for some attributes the magnitude of change was greater for Formulation 1 than for Formulation 2 or Formulation 3.

[0181] The cIEF data for all formulations showed a steady increase in the total % acidic peaks and a corresponding decrease in the % main peaks. However, the magnitude of change over time was significantly greater for Formulation 1 than for Formulation 2 or Formulation 3, especially from T=0 to 3 months and 3 to 6 months.

[0182] This trend was also seen by SEC, where all formulations showed a steady increase in HMWS% peak and a corresponding decrease in % major components, however, similar to the cIEF data, the magnitude of the increase was similar for formulations 2 and 3 and significantly greater for formulation 1.

[0183] Color scoring for Formulations 2 and 3 showed a slight but significant increase in B, BY, and Y color scores over time. In contrast, Formulation 1 showed a rapid increase in color scoring over time with the 6 month sample receiving the maximum B, BY, and Y color scores.

[0184] No change in pH was observed over 6 months for Formulations 2 and 3. However, Formulation 1 showed a significant decrease in pH between 3 and 6 months.

[0185] Post-translational modifications after 3 months at 5℃, 25℃, and 40℃ Post-translational modifications at T=0 and 3M at 5° C., 25° C., and 40° C. are shown in Tables 10-11. At 5° C. and 25° C., all three formulations showed little or little to minimal change in attribute values ​​relative to T=0, respectively. Within a given temperature, the magnitude of change was similar across all formulations, consistent with protein degradation over time at those temperatures.

[0186] At 40° C., a significant increase in deamidation was observed at anti-EGFR HC Asn 333 / anti-c-Met HC Asn 327 and anti-c-Met HC Asn 55, 59 relative to T=0. A similar trend in isomerization of anti-EGFR HC Asp 99 was observed. The magnitude of the increase in deamidation and isomerization was similar for all three formulations.

[0187] Also at 40° C., formulations 2 and 3 showed significant changes in oxidation values ​​versus T=0, consistent with protein degradation over time at 40° C. However, formulation 1 showed a significant increase in the magnitude of oxidation versus formulations 2 and 3 and their corresponding T=0 values ​​for anti-EGFR HC Met 103, anti-EGFR HC Met 108, anti-EGFR HC Met 260 / anti-c-Met HC Met 254, anti-EGFR HC Met 436 / anti-c-Met HC Met 430, and anti-c-Met LC Trp 32, Trp 35.

[0188] Observations and Conclusions Throughout the study, Formulation 1 showed decreased stability attributes versus Formulations 2 and 3. Changes in pH at accelerated temperature suggested that Formulation 1 (10 mM histidine) had poor buffering capacity for amivantamab at higher concentrations (160 mg / mL).

[0189] Formulations 2 and 3 exhibited similar stability profiles over the course of the study. However, at all test time points and temperatures, Formulation 2 appeared as a slightly milky liquid, whereas Formulation 3 appeared as a clear liquid. Similarly, Formulation 3 exhibited lower turbidity values ​​than Formulation 2 throughout the study.

[0190] [Table 7]

[0191] [Table 8]

[0192] [Table 9]

[0193] [Table 10]

[0194] [Table 11]

[0195] [Table 12]

[0196] [Table 13]

[0197] [Table 14]

[0198] Example 3: Shaking and freeze-thaw testing of a range of polysorbate concentrations This study is conducted to determine the range of polysorbate 80 (PS80) concentrations that stabilize amivantamab against mechanical, interfacial, and freeze / thaw stresses. The study also evaluates the protective properties of polysorbate 80 after storage at 5°C for 12 months.

[0199] Multiple identical sets of test formulation vials are made. Each set contains two vials of each of the test formulations containing concentrations of polysorbate 80 below, at, and above the target (0.06% w / v) PS80 value. The set also contains a test formulation control vial that does not contain PS80. All other formulation components are kept constant (160 mg / mL amivantamab, 30 mM acetic acid, 8.5% sucrose, 1 mg / mL methionine, 20 μg / mL ethylenediaminetetraacetic acid (EDTA), pH 5.7). The formulations are dispensed into 8R vials to a fill volume of 7.1 mL, stoppered, capped, and crimp sealed.

[0200] To establish general study baseline data, one set of vials is tested at the start of the study to serve as the untreated time 0 control (T=0).

[0201] To evaluate the stabilizing effect of polysorbate 80 against mechanical and interfacial stresses, one set of vials is placed horizontally on an orbital shaker and shaken at approximately 250 rpm under ambient room temperature and light conditions for up to 72 hours (T72h shake). A second, corresponding, non-shaken control set of vials is held vertically at ambient room temperature and light conditions for the same period of time (T72h control).

[0202] To evaluate the stabilizing effect of aged polysorbate 80 against mechanical and interfacial stresses, two sets of vials are kept for 12 months at 5° C. One set is shaken for up to 72 hours using the method described above (T12m T72h Shake) and the other set is kept as a control (T12m T72h Control).

[0203] To evaluate the stabilizing effect of polysorbate 80 against freeze / thaw stress, one set of vials is subjected to five freeze / thaw cycles (5×FT), where one cycle is defined as freezing to −70° C. followed by passive thawing at ambient room temperature.

[0204] All test samples are evaluated for stability by color, pH, turbidity, particulate matter (not visible to the naked eye), protein concentration (A280), SE-HPLC, cSDS (reduced), cSDS (non-reduced), cIEF, PS80, potency (EGFR), and efficacy (cMET).

[0205] The shaken control sample without PS80 is expected to show an attribute value profile consistent with the presence of agitation-induced degradation, including an increase in the detection of aggregates. Samples containing very low levels of PS80 are also expected to show a sharp decrease in agitation-induced degradation just above or equal to the attribute value profile seen in the T=0 and corresponding non-shaken control samples. Samples containing low, target, and high levels of PS80 are further expected to exhibit attribute value profiles very similar to the T=0 and corresponding non-shaken control samples. This data demonstrates the ability of PS80 to stabilize amivantamab from mechanical and interfacial stresses.

[0206] Similar results are expected for samples held at 5°C for 12 months prior to shaking stress, and no substantial differences in attribute values ​​are expected following freeze / thaw stress compared to the T=0 control.

[0207] It is expected that there will be no substantial difference in the attributes between the low, target and high polysorbate 80 samples under both shaking and freeze / thaw stress, indicating that stable amivantamab formulated with low, target and high concentration levels of polysorbate 80 protects against mechanical, interfacial and freeze / thaw stress.

[0208] Evaluation of polysorbate 80 containing formulations aged under accelerated conditions against shaking and freeze / thaw stress This study was conducted to evaluate a test formulation of 160 mg / mL amivantamab formulated with polysorbate 80 against mechanical, interfacial, and freeze / thaw stresses when aged under accelerated conditions for 6 months.

[0209] Amivantamab 160 mg / mL in 10 mM acetic acid, 8.5% sucrose, 1 mg / mL methionine, 20 μg / mL EDTA, 0.06% PS-80, pH 5.1 was aliquoted into multiple replicate 30R vials with a fill volume of 15.6 mL per vial. The vials were stoppered, capped, and crimp sealed. To establish common study baseline data, one replicate vial was tested at the start of the study and served as the time 0 control (T=0). The remaining replicate vials were placed in a stable location at accelerated (25°C) conditions for 6 months.

[0210] To assess the stability of the aged formulations to mechanical and interfacial stresses, replicate aged vials were placed horizontally on an orbital shaker and shaken at approximately 250 rpm at ambient room temperature (+ shaking) for up to 72 hours.

[0211] To assess the stability of the aged formulation to freeze / thaw stress, aged replicate vials were subjected to five (5) freeze / thaw cycles (5xFT), with one cycle defined as freezing to -70°C for 24 hours, followed by passive thawing at ambient room temperature for 24 hours.

[0212] The remaining aged replicate vials were used as untreated controls for shaking and freeze / thaw studies.

[0213] All test samples were evaluated for stability by color, pH, turbidity, particulate matter (not visible to the naked eye), protein concentration (A280), SE-HPLC, cSDS (reduced), cSDS (non-reduced), cIEF, PS80, potency (EGFR), and efficacy (cMET).

[0214] The test results are presented below in Table 11A. Comparison of T=0 and untreated control samples aged for 6 months under accelerated conditions showed slight to minimal changes in attributes consistent with protein degradation over 6 months at 25° C. Of particular note, all aged samples showed a comparable decrease in PS80 concentration (0.043-0.044%) vs. T=0 (0.058%).

[0215] Comparison of samples aged for 6 months under accelerated conditions showed no significant differences between exposure to shaking stress, repeated freeze-thaw stress, or untreated controls, demonstrating that 160 mg / mL amivantamab formulated at polysorbate 80 concentration levels of 0.04% to 0.06% protects against mechanical, interfacial, and freeze / thaw stress.

[0216] [Table 15]

[0217] [Table 16]

[0218] Example 4: Shaking and freeze-thawing of a range of polysorbate concentrations in rHuPH20-containing formulations This study is conducted to determine the range of polysorbate 80 (PS80) concentrations that stabilize amivantamab with rHuPH20 from mechanical, interfacial, and freeze / thaw stresses. The study also evaluates the protective properties of polysorbate 80 after storage at 5°C for 12 months.

[0219] Multiple identical sets of test formulation vials are made. Each set contains two vials of each of the test formulations containing concentrations of polysorbate 80 below, at, and above the target (0.06% w / v) PS80 value. The set also contains a test formulation control vial that does not contain PS80. All other formulation components are kept constant (160 mg / mL amivantamab, 30 mM acetic acid, 8.5% sucrose, 1 mg / mL methionine, 20 μg / mL ethylenediaminetetraacetic acid (EDTA), 2000 U / mL rHuPH20, pH 5.7). The formulation is dispensed into 8R vials to a fill volume of 7.1 mL, stoppered, capped, and crimp sealed.

[0220] To establish general study baseline data, one set of vials is tested at the start of the study to serve as the untreated time 0 control (T=0).

[0221] To evaluate the stabilizing effect of polysorbate 80 against mechanical and interfacial stresses, one set of vials is placed horizontally on an orbital shaker and shaken at approximately 250 rpm under ambient room temperature and light conditions for up to 72 hours (T72h shake). A second, corresponding, non-shaken control set of vials is held vertically at ambient room temperature and light conditions for the same period of time (T72h control).

[0222] To evaluate the stabilizing effect of aged polysorbate 80 against mechanical and interfacial stresses, two sets of vials are kept for 12 months at 5° C. One set is shaken for up to 72 hours using the method described above (T12m T72h Shake) and the other set is kept as a control (T12m T72h Control).

[0223] To evaluate the stabilizing effect of polysorbate 80 against freeze / thaw stress, one set of vials is subjected to five freeze / thaw cycles (5×FT), where one cycle is defined as freezing to −70° C. followed by passive thawing at ambient room temperature.

[0224] All test samples are evaluated for stability by color, rHuPH20 activity, pH, turbidity, particulate matter (not visible to the naked eye), protein concentration (A280), SE-HPLC, cSDS (reduced), cSDS (non-reduced), cIEF, PS80, potency (EGFR), and efficacy (cMET). The shaken no PS80 control sample is expected to exhibit an attribute profile consistent with the presence of agitation-induced degradation, including increased detection of aggregates. Samples containing very low levels of PS80 are also expected to exhibit a sharp decrease in agitation-induced degradation just above or equal to the attribute profile seen in the T=0 and corresponding non-shaken control samples. Samples containing low, target, and high levels of PS80 are further expected to exhibit attribute profiles very similar to the T=0 and corresponding non-shaken control samples. This data demonstrates the ability of PS80 to stabilize amivantamab from mechanical and interfacial stresses.

[0225] Similar results are expected for samples held at 5°C for 12 months prior to shaking stress, and no substantial differences in attribute values ​​are expected following freeze / thaw stress compared to the T=0 control.

[0226] It is expected that there will be no substantial difference in the attributes between the low, target and high polysorbate 80 samples under both shaking and freeze / thaw stress, indicating that stable amivantamab formulated with low, target and high concentration levels of polysorbate 80 protects against mechanical, interfacial and freeze / thaw stress.

[0227] Example 5: Formulation robustness development Study design A study is conducted to examine the effect of multifactorial variation of formulation component concentration levels of a bispecific EGFR-cMet antibody drug product held at recommended (5°C) and accelerated (25°C) conditions. The formulation components evaluated are protein concentration, acetic acid concentration, sucrose concentration, polysorbate 80 concentration, EDTA / methionine concentration, and pH level. The lowest and highest test factor concentration values ​​tested are approximately 10-40% lower or higher, respectively, than the target test factor concentration values ​​(see Table 12).

[0228] [Table 17]

[0229] Based on this criteria, statistical software was used to create a multifactorial design of experiment statistical model that specified the number and composition of test formulations required for the study.

[0230] The test formulation is prepared and aliquoted into 8R vials with a fill volume of 7.1 mL. The vials are stoppered, capped and crimp sealed. The vials are placed in a stable location at recommended (5° C.) and accelerated (25° C.) conditions. At the designated time points, samples are taken and assayed.

[0231] Test results Test results for each attribute of the test formulation at the start of the study (time 0), after 12 months at recommended storage conditions (5°C), and after 6 months at accelerated temperature (25°C) are presented in tabular format, reporting the range, mean, and standard deviation of the test formulation for each of the following attributes: cIEF (Main Peak Area %, Sum of Acidic Peaks %, Sum of Basic Peaks %), cSDS (Purity % (Non-reduced), Purity % (Reduced)), SE-HPLC (Aggregates %, Monomer %, Fragments %), Particulate Matter (not visible to the naked eye) (particles / container >= 10 μm, particles / container >= 25 μm), and Turbidity (NTU).

[0232] Analytical results for all formulations held at 5°C for 12 months are expected to demonstrate little change in assay test values ​​indicative of stability. The ability of all formulations with multivariate ranges of excipient concentrations to produce narrow ranges of assay test result values ​​demonstrates the robustness of the formulations within the boundaries and storage conditions tested. Moreover, the full range of values ​​observed per assay in this study is expected to be consistent with the most favorable embodiment of stability when held at 2-8°C.

[0233] Analytical results for all formulations held at accelerated storage conditions (25° C.) for 6 months are also expected to show degradation effects consistent with the stability profile of bispecific EGFR-cMet antibodies exposed to long-term accelerated storage conditions. However, for most outcomes, the magnitude of the effect will be relatively small compared to those seen at 12 months at 5° C. Similarly, the magnitude of the increase in the range of outcome values ​​will also be relatively small, with some of the ranges being comparable to or less than those seen at 12 months at 5° C. This demonstrates that even under accelerated storage conditions, the multivariate range of excipient concentrations will yield relatively consistent results.

[0234] Example 6: Development of robustness of formulations with rHuPH20 Study design A study is conducted to examine the effect of multifactorial variation of formulation component concentration levels of a bispecific EGFR-cMet antibody drug product with rHuPH20 held at recommended (5°C) and accelerated (25°C) conditions. The formulation components evaluated are protein concentration, acetic acid concentration, sucrose concentration, polysorbate 80 concentration, EDTA / methionine concentration, rHuPH20 concentration, and pH. The lowest and highest test factor concentration values ​​tested are approximately 10-50% lower or higher, respectively, than the target test factor concentration values ​​(see Table 13).

[0235] [Table 18]

[0236] Based on this criteria, statistical software was used to create a multifactorial design of experiment statistical model that specified the number and composition of test formulations required for the study.

[0237] The test formulation is prepared and aliquoted into 8R vials with a fill volume of 7.5 mL. The vials are stoppered, capped and crimp sealed. The vials are placed in a stable location at recommended (5° C.) and accelerated (25° C.) conditions. At the designated time points, samples are taken and assayed.

[0238] Test results Test results for each attribute of the test formulation at the start of the study (time 0), after 12 months at recommended storage conditions (5°C), and after 6 months at accelerated temperature (25°C) are presented in tabular format, reporting the range, mean, and standard deviation of the test formulation for each of the following attributes: cIEF (main peak area%, sum of acidic peaks%, sum of basic peaks%), cSDS (purity % (non-reduced), purity % (reduced)), SE-HPLC (aggregates %, monomer %, fragments %), particulate matter (not visible to the naked eye) (particles / container >= 10 μm, particles / container >= 25 μm), turbidity (NTU) rHuPH20 activity (U / mL).

[0239] Analytical results for all formulations held at 5°C for 12 months are expected to demonstrate little change in assay test values ​​indicative of stability. The ability of all formulations with multivariate ranges of excipient concentrations to produce narrow ranges of assay test result values ​​demonstrates the robustness of the formulations within the boundaries and storage conditions tested. Moreover, the full range of values ​​observed per assay in this study is expected to be consistent with the most favorable embodiment of stability when held at 2-8°C.

[0240] Analytical results for all formulations held at accelerated storage conditions (25° C.) for 6 months are also expected to show degradation effects consistent with the stability profile of bispecific EGFR-cMet antibodies exposed to long-term accelerated storage conditions. However, for most outcomes, the magnitude of the effect will be relatively small compared to those seen at 12 months at 5° C. Similarly, the magnitude of the increase in the range of outcome values ​​will also be relatively small, with some of the ranges being comparable to or less than those seen at 12 months at 5° C. This demonstrates that even under accelerated storage conditions, the multivariate range of excipient concentrations will yield relatively consistent results.

[0241] Example 7: Bulk production of formulated pharmaceutical Process Description Processing Solution

[0242] [Table 19]

[0243] Ultrafiltration / Diafiltration (UF / DF) Perform ultrafiltration / diafiltration (UF / DF) to reformulate the amivantamab viral retentate intermediate production solution into a preformulated bulk (pFB) solution consisting of 160 mg / mL amivantamab, 30 mM acetate, 8.5% sucrose, 1 mg / mL L-methionine, pH 5.7. Note that during the UF / DF operation, the acetate concentration of the diafiltration buffer increases from 10 mM to 30 mM due to the Gibbs-Donnan effect. The pH also shifts from 5.2 to 5.7.

[0244] Preparation of Amivantamab Formulated Bulk (FB) Polysorbate 80 (6.0% w / v) and EDTA (2 mg / mL) stock solutions are added to the pFB at a 1:100 dilution to obtain a final concentration of 0.06% (w / v) polysorbate 80 and 20 μg / mL EDTA to obtain a formulated bulk (FB) consisting of 160 mg / mL amivantamab in 30 mM acetic acid, 8.5% (w / v) sucrose, 1 mg / mL L-methionine, 0.06% polysorbate 80, 20 μg / mL EDTA, pH 5.7. The FB solution is then mixed homogeneously. Final filtration of the formulated bulk is achieved using a sterile 0.45 / 0.22 μm filter followed immediately by an in-line 0.22 μm filter.

[0245] Final Bulk Fill After the final filtration, the FB is loaded into polycarbonate Biotainer(s) with a loading volume of 20%-90% of the Biotainer's stated volume.

[0246] Final bulk storage and transportation Storage and transportation conditions for the formulated bulk prior to drug manufacture are 5°C ± 3°C, protected from light, if the FB is stored for approximately 1 week or less, or -40°C ± 10°C, protected from light, if the FB is stored for more than 1 week.

[0247] Example 8: Pharmaceutical Formulations with rHuPh20 Products Process Description Processing Solution

[0248] [Table 20] 1) 100,000IU / mg rHuPH20

[0249] Preparation of Amivantamab Pharmaceutical Formulation with rHuPH20 The rHuPH20 formulation bulk is added to the amivantamab formulation bulk solution to obtain a final concentration of 2,000 IU / mL of rHuPH20, resulting in a pharmaceutical formulation with rHuPH20 consisting of 30 mM acetic acid, 8.5% (w / v) sucrose, 1 mg / mL L-methionine, 0.06% polysorbate 80, 20 μg / mL EDTA, rHuPH20 2000 IU, 160 mg / mL amivantamab at pH 5.7. The contribution of histidine and sodium chloride from the rHuPH20 formulation bulk is believed to be minimal and therefore is not listed in the amivantamab containing rHuPH20 pharmaceutical formulation composition. The pharmaceutical formulation solution is then mixed homogeneously. An initial filtration of the pharmaceutical formulation is achieved using a sterile 0.22 μm filter. Final filtration of the pharmaceutical formulation is accomplished using a sterile 0.22 μm filter followed immediately by an in-line 0.22 μm filter.

[0250] Example 9: Pharmaceuticals: Composition and ingredients of primary packaging A summary of the composition of the amivantamab drug product is provided herein in tabular form (Table 16).

[0251] [Table 21]

[0252] The amivantamab drug product (DP) primary packaging consists of a glass vial, a polymeric vial stopper, and an aluminum seal. Table 17 lists the specific components of the primary packaging materials.

[0253] [Table 22]

[0254] Example 10: Pharmaceuticals with rHuPH20: Composition and Components of Primary Packaging The composition summary of the amivantamab drug product with rHuPH20 is provided herein in tabular form (Table 18).

[0255] [Table 23]

[0256] The amivantamab drug product (DP) primary packaging with rHuPH20 consists of a glass vial, a polymeric vial stopper, and an aluminum seal. Table 19 lists the specific components of the primary packaging materials.

[0257] [Table 24]

[0258] Example 11: Drug Stability Testing Description This study was conducted to monitor stable site amivantamab DP attributes under various environmental conditions and time lengths. Investigational test articles were prepared by aliquoting the formulated bulk into 8R vials at 7.1 mL fill volumes. The vials were stoppered, capped, and crimp sealed.

[0259] All tests were performed with the vials inverted.

[0260] [Table 25]

[0261] Stability test results Stability results to date for amivantamab DP held under recommended, accelerated, and stressed conditions are listed in Tables 21-23. At all time points for the DP held at recommended storage conditions, all test parameter result values ​​observed per assay study are expected to exceed criteria consistent with the most preferred embodiment of stability when held after about 12 months or more of storage at a temperature of about 5° C., after about 12 months or more of storage at a temperature of about 25° C., and / or after about 2 years or more of storage at a temperature of about 5° C. Similarly, peptide map results are expected to show little or no consequential change over time in the measured rates of post-translational modifications.

[0262] The results for the amivantamab DP held under accelerated and stressed conditions are expected to demonstrate the expected degradation rates of drug products exposed to long-term accelerated and stressed storage conditions. The DP held at accelerated conditions (25°C) for 12 months is also expected to demonstrate results consistent with preferred embodiments of stability when held at 2-8°C for approximately 2 years or more.

[0263] 5℃ data

[0264] [Table 26]

[0265] [Table 27]

[0266] [Table 28]

[0267] [Table 29]

[0268] [Table 30] NT = not tested

[0269] [Table 31]

[0270] 25℃ data

[0271] [Table 32]

[0272] [Table 33]

[0273] [Table 34]

[0274] [Table 35]

[0275] [Table 36] NT = not tested

[0276] [Table 37]

[0277] [Table 38]

[0278] [Table 39]

[0279] [Table 40]

[0280] [Table 41]

[0281] [Table 42] NT = not tested

[0282] [Table 43]

[0283] Example 12: Description of Drug Stability Studies with rHuPH20 This study was conducted to monitor amivantamab with rHuPH20 DP attributes in a stable location under various environmental conditions and time lengths. Investigational test articles were prepared by aliquoting the formulated bulk into 8R vials at 7.1 mL fill volumes. The vials were stoppered, capped, and crimp sealed.

[0284] All tests were performed with the vials inverted.

[0285] [Table 44]

[0286] Stability test results Stability results to date for amivantamab with rHuPH20 DP held under recommended, accelerated, and stressed conditions are listed in Tables 25-27. At all time points for the DP held at recommended storage conditions, all test parameter result values ​​observed per assay study are expected to exceed criteria consistent with the most preferred embodiment of stability when held at a temperature of about 5° C. for about 12 months or more, at a temperature of about 25° C. for about 12 months or more, and / or at a temperature of about 5° C. for about 2 years or more. Similarly, peptide map results are expected to show little or no consequential change over time in the measured rates of post-translational modifications.

[0287] The results for amivantamab with the rHuPH20 DP held under accelerated and stressed conditions are expected to demonstrate the expected degradation rates of drug products exposed to long-term accelerated and stressed storage conditions. It is also expected that the DP held at accelerated conditions (25°C) for 12 months will demonstrate results consistent with preferred embodiments of stability when held at 2-8°C for approximately 2 years or more.

[0288] 5℃ data

[0289] [Table 45]

[0290] [Table 46]

[0291] [Table 47]

[0292] [Table 48]

[0293] [Table 49] NT = not tested

[0294] [Table 50]

[0295] 25℃ data

[0296] [Table 51]

[0297] [Table 52]

[0298] [Table 53]

[0299] [Table 54]

[0300] [Table 55] NT = not tested

[0301] [Table 56]

[0302] 40℃ data

[0303] [Table 57]

[0304] [Table 58]

[0305] [Table 59]

[0306] [Table 60]

[0307] [Table 61] NT = not tested

[0308] [Table 62]

[0309] Example 12A: Viscosity The viscosity of 160 mg / mL amivantamab, 30 mM acetate, 8.5% sucrose, 0.06% polysorbate 80, 20 μg / mL EDTA, 1 mg / mL methionine at pH 5.7 was analyzed with an automated viscometer. Temperature sweeps were performed between 4° C. and 40° C. at 5° C. intervals, with four measurements taken at each temperature setting. The first measurement at every temperature was discarded and the remaining three were used to calculate the average viscosity values ​​reported in Table 28 below.

[0310] [Table 63]

[0311] These results demonstrate that the highly concentrated formulation of amivantamab has a suitable viscosity for injection.

[0312] Example 13. Subcutaneous delivery of amivantamab in patients with advanced solid malignancies. Subcutaneous (SC) delivery of amivantamab was evaluated in a Phase 1 dose escalation study in patients with advanced solid tumors that could benefit from EGFR- or MET-directed therapy (PALOMA; NCT04606381). Eligible tumor types included non-small cell lung cancer (NSCLC), squamous cell carcinoma of the head and neck (SCCHN), hepatocellular carcinoma (HCC), colorectal cancer (CRC), renal cell carcinoma (RCC), medullary thyroid carcinoma (MTC), gastroesophageal carcinoma (GEC), mesothelioma, breast cancer (BC), and ovarian cancer (OC). Eligible patients had progressed after standard treatment for metastatic disease and were ineligible for or had refused current standard treatment.

[0313] The study objective was to evaluate the administration, safety, and pharmacokinetics (PK) feasibility of a low-concentration formulation, 50 mg / mL amivantamab ± rHuPH20 (part 1) and a high-concentration formulation, 160 mg / mL amivantamab ± rHuPH20 (part 2). A low-concentration formulation (50 mg / mL) amivantamab was administered either in (Ami-LC-MD [mixed and delivered]) or without (Ami-LC) rHuPH20 (part 1, cohorts 1a and 1b, respectively). A high-concentration formulation (160 mg / mL) amivantamab was administered either in (Ami-HC-CF [co-formulated]) or without (Ami-HC) rHuPH20 (part 2, cohorts 2a and 2b, respectively). Patients in parts 1 and 2 received amivantamab doses of 1050 mg and 2,000 units / mL rHuPH20 (1400 mg and 2000 units / mL rHuPH20 for those weighing ≥ 80 kg) SC (weekly for the first 4 weeks, then every other week). The study also evaluated administration of a full dose of amivantamab on day 1.

[0314] Results: Complete safety, PK, bioavailability, and receptor occupancy data were evaluated for patients enrolled in Part 1 (n=16) and Part 2 (n=17). Compared with IV dosing, initial SC experience demonstrated that coformulation of high-concentration amivantamab with rHuPH20 reduced the required infusion time from 2-4 hours to less than 5 minutes, with initial bioavailability approximately 65% ​​of that of IV dosing. Saturation of soluble free EGFR and MET was achieved after the first SC dose. The incidence of IRR was 18.2%, compared with 67.3% of patients receiving IV amivantamab at the recommended phase 2 dose (RP2D) across the CHRYSALIS study (Park Ann Oncol 32[suppl_5]:S981), with all events being grade 1 to grade 2 in severity. The full amivantamab SC dose was safely administered at first dose to 14 patients, eliminating the need for split dosing.

[0315] No increased risk of IRR was observed with full initial amivantamab dosing; IRR was reported in 3 / 14 (21%) patients receiving a full dose (C1D1) and in 3 / 19 (16%) patients receiving split doses.

[0316] The AE profile outside the IRR for SC amivantamab was comparable to IV amivantamab. Grade ≥3 AEs occurred in 27.3% of patients, mostly reported as single events. A single grade 3 event (hypokalemia) was reported as treatment-related. No treatment-related AEs leading to dose reduction or discontinuation were reported. Two patients (6.1%) had grade 1 infusion site reactions, which were transient and did not affect subsequent dosing.

[0317] Pharmacokinetics, Pharmacodynamics, and Immunogenicity. Maximum serum concentrations of amivantamab were achieved 2–3 days after SC administration (Figure 1). Higher exposure was obtained with rHuPH20; estimated bioavailability with SC administration using formulations containing rHuPH20 was approximately 65%. Saturation of soluble free EGFR and MET was achieved in all cohorts after the first full dose (Figure 2). No anti-drug antibodies were detected in patients studied (total N=33 in both Part 1 and Part 2 arms).

[0318] Conclusions: Initial SC amivantamab ± rHuPH20 was well tolerated with improved time and ease of administration, and was associated with a significant reduction in IRR compared with intravenous (IV) administration, eliminating the need for split dosing.

[0319] Example 14. Amivantamab and lazertinib in patients with EGFR-mutated non-small cell lung (NSCLC) after progression on osimertinib and platinum-based chemotherapy. Methods: Clinical trial cohorts evaluated amivantamab and lazertinib in patients with EGFR exon 19 deletion or L858R NSCLC with disease that progressed after 1st / 2nd line osimertinib followed by platinum-chemotherapy as the last line of treatment (target population, n=106), and in a more heavily pretreated population (n=56) with disease that progressed after osimertinib and platinum-chemotherapy ± other therapies regardless of the number and sequence of these therapies. Patients received 1050 mg IV amivantamab (1400 mg, ≥80 kg) + 240 mg oral lazertinib. Responses were assessed according to RECIST v1.1 (European Journal of Cancer, vol. 45, pp. 228-247 (2009)) and reported for response-evaluable patients, defined as patients with ≥6 months follow-up for response durability.

[0320] Results: 162 patients were enrolled in the cohort (median age 62 years on prior line of therapy, 65% female, 61% Asian, median 3 [range, 2-14]). Median time from last osimertinib treatment to first dose of amivantamab plus lazertinib was 6.3 and 2.0 months in the target and heavily pretreated populations, respectively. Among 50 efficacy-evaluable patients in the target population, ORR was 36% (95% CI, 23-51), with 1 complete response (CR) and 17 partial responses (PR), for a clinical benefit rate (CBR) of 58% (95% CI, 43-72). Median duration of response (mDOR) was not reached. At a median follow-up of 8.3 months, 7 responders (39%) achieved a DOR lasting ≥6 months. Among 56 efficacy-evaluable patients in the heavily pretreated population (median follow-up of 8.7 months), the ORR was 29% (95% CI, 17-42), with 1 CR and 15 PR. CBR was 55% (95% CI, 42-69), and mDOR was 8.6 months (95% CI, 4.2-NR). Preliminary evidence of CNS antitumor activity was reported in 8 patients with baseline brain lesions (7 non-targeted, 1 targeted) who had not received radiation within 1 year prior to study enrollment.

[0321] Infusion-related reactions (IRR) (65%) were the most common adverse events (AEs) reported, followed by paronychia (49%), rash (41%), and stomatitis (39%). The most common grade ≥ 3 treatment-related AEs (TRAEs) were infusion-related reactions (7%), dermatitis acneiform (5%), and hypoalbuminemia (4%). TRAEs leading to discontinuation of either or both amivantamab (ami) and lazertinib (laz) occurred in 12% and 7%, respectively.

[0322] Example 15. Subcutaneous delivery of amivantamab co-formulated with rHuPH20. The proposed preliminary recommended phase 2 dose (RP2D) for the once every 2 weeks (Q2W) amivantamab dosing regimen is 1600 mg for patients with BW < 80 kg and 2240 mg for participants with BW ≥ 80 kg. This preliminary subcutaneous dose contains amivantamab at a concentration of 160 mg / mL co-formulated with rHuPH20 (amivantamab SC-CF).

[0323] The proposed recommended dosing regimen for amivantamab SC-CF is chosen to ensure that the resulting exposure is similar to that observed with the IV RP2D regimen. Bioavailability was estimated by comparing the observed AUC after SC dosing (cohorts with and without rHuPH20 formulation) with the corresponding observed AUC after IV dosing. The SC cohort was administered the IV RP2D (1050 mg for participants with BW<80 kg and 1400 mg for participants with BW>=80 kg). At this dose, saturation of soluble free EGFR and cMet was achieved after the first SC dose. Additionally, all 33 participants were negative for anti-amivantamab antibodies. After the first dose of cycle 2 (after the weekly induction dose in cycle 1), the mean (CV%, n) area under the concentration-time curve (AUC) C2D1–C2D15 was 95,416 μg×h / mL (45.4%, 7) and 75,378 μg×h / mL (27.0%, 5) for cohorts from 160 mg / ml with and without rHuPH20, respectively.

[0324] Coformulation of amivantamab with rHuPH20 (i.e., amivantamab SC-CF) provided improved bioavailability (65% vs. 51%) in addition to reducing the time required for infusion by approximately 8-fold compared with formulations without rHuPH20. Based on estimated bioavailability, a preliminary RP2D of 1,600 mg for participants with BW < 80 kg and 2,240 mg for participants with BW ≥ 80 kg was proposed.

[0325] Emerging data show a lower incidence of IRR (18.7% overall and 0 grade ≥3) than previously reported with IV amivantamab (65.9% overall and 2.3% grade ≥3). As a result of the reduced incidence and severity of IRR, this study demonstrated the feasibility of a daily infusion of the initial dose of amivantamab SC.

[0326] Based on current clinical data and analysis supported by preliminary modeling, amivantamab SC-CF will be administered at a preliminary RP2D of 1,600 mg for participants weighing less than 80 kg and 2,240 mg for participants weighing more than 80 kg.

[0327] Cohort 1 evaluates amivantamab SC-CF (Q2W) in combination with lazertinib in subjects with treatment-naïve locally advanced or metastatic NSCLC harboring EGFR exon 19del or exon 21 L858R mutations. Participants will receive amivantamab on days 1, 8, 15, and 22 of cycle 1 and days 1 and 15 of each 28-day cycle thereafter, starting with cycle 2. Amivantamab SC-CF will be administered SC by manual injection at 1,600 mg (2,240 mg if body weight ≥ 80 kg). Lazertinib will be administered orally at 240 mg once daily.

[0328] Cohort 4 will evaluate the feasibility of amivantamab SC-CF (Q2W) as switch therapy in subjects who have received 3 or more months of amivantamab IV according to standard of care. Participants will receive amivantamab on days 1, 8, 15, and 22 of cycle 1 and days 1 and 15 of each 28-day cycle thereafter, starting with cycle 2. Amivantamab SC-CF will be administered by manual injection at 1,600 mg (2,240 mg if body weight ≥ 80 kg).

[0329] Those skilled in the art will appreciate that numerous changes and modifications may be made to the preferred embodiments of the present invention, and that such changes and modifications may be made without departing from the spirit of the present invention. It is therefore intended in the appended claims to cover all such equivalent variations that fall within the true spirit and scope of the present invention.

[0330] The disclosures of each patent, patent application, and publication cited or described in this specification are hereby incorporated by reference in their entirety.

[0331] [Table 64]

[0332] [Table 65-1]

[0333] [Table 65-2]

Claims

1. 1. A stable aqueous pharmaceutical composition comprising a bispecific epidermal growth factor receptor (EGFR) / hepatocyte growth factor receptor (c-Met) antibody and hyaluronidase, wherein the antibody is a. a first heavy chain (HC1) comprising a HC1 variable region (VH1) comprising heavy chain complementarity determining region 1 (HCDR1), HCDR2, and HCDR3 comprising the amino acid sequences of SEQ ID NOs: 1, 2, and 3, respectively; b. a first light chain (LC1) comprising an LC1 variable region (VL1) comprising light chain complementarity determining region 1 (LCDR1), LCDR2, and LCDR3 comprising the amino acid sequences of SEQ ID NOs: 4, 5, and 6, respectively; c. a second heavy chain (HC2) comprising a HC2 variable region (VH2) comprising HCDR1, HCDR2, and HCDR3 comprising the amino acid sequences of SEQ ID NOs: 7, 8, and 9, respectively; d. a second light chain (LC2) comprising an LC2 variable region (VL2) comprising LCDR1, LCDR2, and LCDR3 comprising the amino acid sequences of SEQ ID NOs: 10, 11, and 12, respectively; A stable aqueous pharmaceutical composition comprising:

2. The stable aqueous pharmaceutical composition according to claim 1, a) about 144 mg / mL to about 176 mg / mL of a bispecific epidermal growth factor receptor (EGFR) / hepatocyte growth factor receptor (c-Met) antibody; b) about 10 mM to about 50 mM acetic acid and / or a pharmaceutically acceptable acetate salt; c) about 6.8% (w / v) to about 10.2% (w / v) sucrose; and d) about 0.036% (w / v) to about 0.084% (w / v) of polysorbate 80 (PS80); and e) about 0.8 mg / mL to about 1.2 mg / mL methionine; f) about 16 μg / mL to about 24 μg / mL ethylenediaminetetraacetic acid (EDTA); g) optionally, about 1,000 U / mL to about 3,000 U / mL of hyaluronidase; h) a pH of about 5.2 to about 6.2; and A stable aqueous pharmaceutical composition comprising:

3. 2. The stable aqueous pharmaceutical composition of claim 1, wherein the bispecific EGFR / c-Met antibody comprises an HC1 variable region comprising the amino acid sequence of SEQ ID NO: 13, an LC1 variable region comprising the amino acid sequence of SEQ ID NO: 14, an HC2 variable region comprising the amino acid sequence of SEQ ID NO: 15, and an LC2 variable region comprising the amino acid sequence of SEQ ID NO:

16.

4. 2. The stable aqueous pharmaceutical composition of claim 1, wherein HC1 comprises the amino acid sequence of SEQ ID NO: 17, LC1 comprises the amino acid sequence of SEQ ID NO: 18, HC2 comprises the amino acid sequence of SEQ ID NO: 19, and LC2 comprises the amino acid sequence of SEQ ID NO:

20.

5. 2. The stable aqueous pharmaceutical composition of claim 1, wherein the bispecific EGFR / c-Met antibody is amivantamab or a biosimilar thereof.

6. 3. The stable aqueous pharmaceutical composition of claim 2, wherein the bispecific EGFR / c-Met antibody has a concentration of about 160 mg / mL.

7. 3. The stable aqueous pharmaceutical composition of claim 2, wherein the acetic acid and / or pharmaceutically acceptable acetate salt has a concentration of about 30 mM.

8. 3. The stable aqueous pharmaceutical composition of claim 2, wherein the acetic acid and / or pharmaceutically acceptable acetate salt comprises glacial acetic acid and / or sodium acetate trihydrate.

9. 3. The stable aqueous pharmaceutical composition of claim 2, comprising about 8.5% (w / v) sucrose.

10. 3. The stable aqueous pharmaceutical composition of claim 2, comprising about 0.06% (w / v) PS80.

11. 3. The stable aqueous pharmaceutical composition of claim 2, wherein the methionine comprises L-methionine and has a concentration of about 1 mg / mL.

12. 3. The stable aqueous pharmaceutical composition of claim 2, wherein the EDTA has a concentration of about 20 μg / mL.

13. 3. The stable aqueous pharmaceutical composition of claim 2, wherein the pH is about 5.

7.

14. 3. The stable aqueous pharmaceutical composition of claim 2, wherein the hyaluronidase is a human hyaluronidase, optionally a soluble human PH20 comprising the amino acid sequence of SEQ ID NOs: 21-25.

15. 3. The stable aqueous pharmaceutical composition of claim 2, wherein the composition comprises rHuPH20 at a concentration of about 1,000 U / mL to about 3,000 U / mL.

16. 3. The stable aqueous pharmaceutical composition of claim 2, wherein the hyaluronidase is human recombinant hyaluronidase enzyme PH20 (rHuPH20) at a concentration of about 2,000 U / mL.

17. the stable aqueous pharmaceutical composition comprising about 160 mg / mL of the bispecific EGFR / c-Met antibody, about 30 mM acetic acid and / or a pharmaceutically acceptable acetate salt, about 8.5% sucrose, and about 1 mg / mL L-methionine, together with polysorbate 80 to a final concentration of about 0.06% (w / v) and EDTA to a final concentration of about 20 μg / mL, wherein the stable aqueous pharmaceutical composition has a pH of about 5.7; the bispecific EGFR / c-Met antibody comprises a heavy chain 1 (HC1) comprising the amino acid sequence of SEQ ID NO: 17, a HC2 comprising the amino acid sequence of SEQ ID NO: 19, a light chain 1 (LC1) comprising the amino acid sequence of SEQ ID NO: 18, and a LC2 comprising the amino acid sequence of SEQ ID NO:

20. The stable aqueous pharmaceutical composition according to claim 2.

18. the stable aqueous pharmaceutical composition comprising about 160 mg / mL of the bispecific EGFR / c-Met antibody, about 30 mM acetic acid and / or a pharmaceutically acceptable acetate salt, about 8.5% sucrose, about 1 mg / mL L-methionine, together with polysorbate 80 to a final concentration of about 0.06% (w / v), EDTA to a final concentration of about 20 μg / mL, and human recombinant hyaluronidase enzyme PH20 (rHuPH20) to a final concentration of about 2,000 U / mL, wherein the stable aqueous pharmaceutical composition has a pH of about 5.7; the bispecific EGFR / c-Met antibody comprises a heavy chain 1 (HC1) comprising the amino acid sequence of SEQ ID NO: 17, a HC2 comprising the amino acid sequence of SEQ ID NO: 19, a light chain 1 (LC1) comprising the amino acid sequence of SEQ ID NO: 18, and a LC2 comprising the amino acid sequence of SEQ ID NO:

20. The stable aqueous pharmaceutical composition according to claim 2.

19. 10. The pharmaceutical composition of claim 1 for use in a method of treating cancer in a subject in need thereof, comprising: The method comprises administering the pharmaceutical composition to the subject.

20. 20. The pharmaceutical composition of claim 19, wherein the administration is subcutaneous.

21. 20. The pharmaceutical composition of claim 19, wherein the cancer comprises lung cancer, squamous cell carcinoma of the head and neck (SCCHN), hepatocellular carcinoma (HCC), colorectal cancer (CRC), renal cell carcinoma (RCC), medullary thyroid carcinoma (MTC), gastroesophageal carcinoma (GEC), mesothelioma, breast cancer (BC), or ovarian cancer (OC).

22. 22. The pharmaceutical composition of claim 21, wherein the cancer comprises non-small cell lung cancer (NSCLC).

23. The pharmaceutical composition of claim 21, wherein the cancer comprises colorectal cancer (CRC).

24. The pharmaceutical composition of claim 21, wherein the cancer comprises squamous cell carcinoma of the head and neck (SCCHN).

25. A method for preparing a stable aqueous pharmaceutical composition of a bispecific antibody targeting EGFR and c-Met, wherein the bispecific antibody targeting EGFR and c-Met comprises a first heavy chain (HC1) comprising an HC1 variable region (VH1) comprising heavy chain complementarity determining region 1 (HCDR1), HCDR2, and HCDR3 comprising the amino acid sequences of SEQ ID NOs: 1, 2, and 3, respectively, and a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequences of SEQ ID NOs: 4, 5, and 6, respectively. a first light chain (LC1) comprising an LC1 variable region (VL1) comprising an LCDR1, an HCDR2, and an HCDR3 comprising the amino acid sequences of SEQ ID NOs: 7, 8, and 9, respectively; and a second light chain (LC2) comprising an LC2 variable region (VL2) comprising an LCDR1, an LCDR2, and an LCDR3 comprising the amino acid sequences of SEQ ID NOs: 10, 11, and 12, respectively, wherein the method comprises:

10. A method comprising combining a composition comprising about 160 mg / mL of the bispecific antibody, about 30 mM acetic acid and / or a pharmaceutically acceptable acetate salt, about 8.5% sucrose, and about 1 mg / mL L-methionine with polysorbate 80 to a final concentration of about 0.06% (w / v) and EDTA to a final concentration of about 20 μg / mL, optionally with rHuPH20 to a final concentration of about 2,000 U / mL, wherein the stable aqueous pharmaceutical composition has a pH of about 5.

7.

26. 26. The method of claim 25, wherein the bispecific EGFR / c-Met antibody comprises an HC1 variable region comprising the amino acid sequence of SEQ ID NO: 13, an LC1 variable region comprising the amino acid sequence of SEQ ID NO: 14, an HC2 variable region comprising the amino acid sequence of SEQ ID NO: 15, and an LC2 variable region comprising the amino acid sequence of SEQ ID NO:

16.

27. 26. The method of claim 25, wherein the antibody comprises a heavy chain 1 (HC1) comprising the amino acid sequence of SEQ ID NO: 17, a light chain 1 (LC1) comprising the amino acid sequence of SEQ ID NO: 18, a HC2 comprising the amino acid sequence of SEQ ID NO: 19, and a LC2 comprising the amino acid sequence of SEQ ID NO:

20.

28. 26. The method of claim 25, wherein the antibody is amivantamab or a biosimilar thereof.

29. A kit comprising the stable aqueous pharmaceutical composition of claim 1 and instructions for use thereof.

30. 10. An article of manufacture comprising a container holding the stable aqueous pharmaceutical composition of claim 1.

31. 31. The article of manufacture of claim 30, wherein the container is a vial having a stopper pierceable by a syringe.

32. 32. The article of manufacture of claim 31 , wherein the vial is a single-use vial.

33. 10. The pharmaceutical composition of claim 1 for use in a method for reducing infusion-related reactions in a subject treated with amivantamab, comprising: The method comprises subcutaneously administering the stable aqueous pharmaceutical composition to a subject.

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