RU2807524C2 - Compositions of antibodies to rsv and methods of their application - Google Patents

Abstract

FIELD: RSV antibody compositions.

SUBSTANCE: present invention is related to stable compositions containing antibodies or antigen-binding fragments thereof that bind to respiratory syncytial virus (RSV). Methods of preventing and/or treating RSV-related diseases using the compositions of the present invention are also provided.

EFFECT: invention provides formulations that contain a high concentration of antibody or antigen-binding fragment thereof and a viscosity level suitable for subcutaneous delivery, and do not result in high levels of aggregation.

20 cl, 15 dwg, 10 tbl, 8 ex

Description

Field of technology to which the present invention relates

The present invention relates to stable compositions containing antibodies or antigen-binding fragments thereof that bind to respiratory syncytial virus (RSV). Methods of preventing and/or treating RSV-related diseases using the compositions of the present invention are also provided.

Link to related applications

Not applicable.

Link to electronically submitted sequence listing

The sequence listing of this application is provided electronically via EFS-Web as a sequence listing in ASCII format with the file name "24641WOPCT-SEQLIST.TXT", creation date October 9, 2019, and size 9.19 kB. This sequence listing provided via EFS-Web constitutes part of the specification and is incorporated herein by reference in its entirety.

BACKGROUND OF THE INVENTION

Paramyxoviruses are enveloped negative-strand RNA viruses that are important pathogens of humans and animals. Human respiratory syncytial virus (hRSV, RSV) belongs to the family Paramyxoviridae, subfamily Pneumovirinae. Two subtypes, type A and type B, have been identified and represent the leading cause of severe and sometimes fatal respiratory infections in children under 6 months of age. Adults with underlying medical conditions, such as COPD, asthma, and cancer, and adults who are immunocompromised, including HIV or post-transplant, are also at risk for developing severe RSV infection. Fifteen percent of annual hospitalizations in adults over 50 years of age due to acute respiratory infection are caused by RSV. In the United States, RSV causes more than 100,000 hospitalizations and is estimated to cause approximately 160,000 deaths annually worldwide. There is currently no vaccine against RSV, and a trial with formalin-inactivated virus was associated with increased severity of illness in infants following RSV infection. Other members of the family, including human metapneumovirus (hMPV) and human parainfluenza virus (hPIV), are also responsible for acute respiratory disease similar to hRSV.

The hRSV genome is a single-stranded negative-sense RNA molecule of approximately 15 kb in size that encodes 11 proteins. Two of these proteins are the major surface glycoproteins of the virion. These are (i) the attachment protein (G), which mediates the binding of the virus to cells, and (ii) the fusion protein (F), which promotes both the fusion of the viral and cellular membranes at the initial stages of the infectious cycle, and the fusion of the membrane of infected cells with the membranes of neighboring cells with the formation of characteristic syncytia. Attachment protein G binds to cell surface receptors and interacts with protein F. This interaction triggers a conformational change in protein F to cause membrane fusion, thereby releasing the viral ribonucleoprotein complex into the cytoplasm of the host cell.

Monoclonal antibodies to the F or G protein have been shown to have neutralizing effects in vitro and prophylactic effects in vivo. See, for example, Beeler and Coelingh 1989, J. Virol. 63:2941-50; Garcia-Barreno et al, 1989, J. Virol. 63:925-32; Taylor et al, 1984, Immunology 52:137-142; Walsh et al, 1984, Infection and Immunity 43:756-758 and US Patent Nos. 5842307 and 6818216. Other antibodies to hRSV are described in international patent applications WO 94/06448 and WO 92/04381, as well as in US patents 8221759 and 9963500.

Antibodies for use in human subjects must be stored until use and transported to the site of administration. Reproducibly achieving the desired level of an antibody drug in a subject requires that the drug be stored in a formulation that maintains the bioactivity of the drug. There is a need for stable anti-RSV antibody formulations for pharmaceutical use, for example, for the prevention and/or treatment of RSV-related diseases. Preferably, such formulations will have a long shelf life, be stable during storage and transportation, and can be administered in high concentrations, for example for subcutaneous administration, as well as in low concentrations, for example, for intravenous administration.

The compositions of the present invention are useful for subcutaneous delivery to a patient in need thereof. To provide maximum therapeutic benefit to patients, it is desirable that formulations for subcutaneous (SC) delivery contain a high concentration of antibodies (75-200 mg/mL). For subcutaneous injections, a high concentration of API is often required due to the historical bioavailability of 50-60% for subcutaneous injections and the expected dosage range of the antibody product. However, a high concentration of antibody or antigen-binding fragment thereof may contribute to other properties of the product that would be undesirable, such as low injectability due to increased viscosity and higher physiological osmolality and increased aggregation. Therefore, it is preferable that the antibody product intended for subcutaneous administration balances the concentration effects while maintaining the drug level that provides the highest therapeutic effect. The ideal product contains high protein concentration, low viscosity, osmolality similar to physiological conditions, and low levels of aggregation under typical storage conditions. Increased viscosity at high protein concentrations can not only make it difficult to remove the product from the container using a syringe, but also make it difficult to administer the required dose from the syringe to the patient (needle patency). Advantageously, embodiments of the present invention provide formulations that contain a high concentration of antibody or antigen-binding fragment thereof and a viscosity level suitable for subcutaneous delivery. In addition, the compositions of the present invention do not result in high levels of aggregation, as shown in more detail in the examples.

Summary of the present invention

The present disclosure provides an anti-RSV antibody composition comprising: a) about 50 mg/ml to about 250 mg/ml of an anti-RSV antibody or antigen-binding fragment thereof; b) about 5 mM to about 20 mM buffer; c) a stabilizer selected from the group consisting of: (i) from about 6% to about 8% in a mass-volume ratio (w/v) of non-reducing sugar; (ii) from about 25 mm to about 75 mm L-arginine or a pharmaceutically acceptable salt of L-arginine, L-proline or a pharmaceutically acceptable salt of L-proline; L-lysine or a pharmaceutically acceptable salt of L-lysine; L-glutamate or a pharmaceutically acceptable salt of L-glutamate or a mixture of amino acids described herein, and (iii) about 25 μM to 75 μM chelator; d) about 0.01% to about 0.10% nonionic surfactant; and e) about 1 mM to about 20 mM antioxidant.

In another embodiment, the present disclosure provides an anti-RSV antibody composition comprising: a) an anti-RSV antibody or antigen-binding fragment thereof at a concentration of about 50 mg/ml to about 150 mg/ml; b) histidine at a concentration of about 5 mM to about 20 mM; c) sucrose at a concentration of from about 6% to about 8% in a mass-volume ratio (w/v); d) an excipient selected from the following: L-arginine or a pharmaceutically acceptable L-arginine salt at a concentration of about 25 mM to about 75 mM, L-lysine or a pharmaceutically acceptable L-lysine salt at a concentration from about 25 mM to about 75 mM , a mixture of L-arginine at a concentration of about 25 mM to about 75 mM and L-glutamate at a concentration of about 25 mM to about 75 mM and a mixture of L-arginine (HCl) at a concentration of about 25 mM to about 75 mM and L- lysine (HCl) at a concentration of about 25 mM to about 75 mM; e) DTPA at a concentration of about 25 μM to 75 μM; f) polysorbate 80 at a concentration of about 0.01% to about 0.10%; and optionally g) L-methionine at a concentration of about 1 mM to about 20 mM.

In another embodiment, the present disclosure provides an anti-RSV antibody composition comprising: a) an anti-RSV antibody or antigen binding fragment thereof at a concentration of about 100 mg/ml; b) histidine at a concentration of about 10 mM; c) sucrose at a concentration of about 7% in a mass-volume ratio (w/v); d) an excipient selected from L-arginine (HCl) at a concentration of about 50 mM, L-lysine (HCl) at a concentration of about 50 mM, a mixture of L-arginine at a concentration of about 25 mM and L-glutamate at a concentration of about 25 mM, and mixtures of L-arginine (HCl) at a concentration of about 25 mM and L-lysine (HCl) at a concentration of about 25 mM; e) DTPA at a concentration of about 50 μM; f) polysorbate 80 at a concentration of about 0.02%; and optionally g) L-methionine at a concentration of about 1 mM to 20 mM.

In another embodiment, the present disclosure provides an anti-RSV antibody composition comprising: a) an anti-RSV antibody or antigen binding fragment thereof at a concentration of about 100 mg/ml; b) histidine at a concentration of about 10 mM; c) sucrose at a concentration of about 7% in a mass-volume ratio (w/v); d) an excipient selected from L-arginine (HCl) at a concentration of 50 mM, L-lysine (HCl) at a concentration of 50 mM, a mixture of L-arginine at a concentration of 25 mM and L-glutamate at a concentration of 25 mM and a mixture of L-arginine at a concentration of 25 mM (HCl) and L-lysine (HCl) at a concentration of 25 mM; f) DTPA at a concentration of 50 μM; f) polysorbate 80 at a concentration of 0.02% and optionally g) L-methionine at a concentration of 1 mM to 20 mM.

In another embodiment, the present disclosure provides an anti-RSV antibody composition comprising: a) an anti-RSV antibody or antigen binding fragment thereof at a concentration of about 100 mg/ml; b) histidine at a concentration of about 10 mM; c) sucrose at a concentration of about 7% in a mass-volume ratio (w/v); d) a mixture of L-arginine at a concentration of 25 mM (HCl) and L-lysine (HCl) at a concentration of 25 mM; e) DTPA at a concentration of 50 µM and f) polysorbate 80 at a concentration of 0.02%.

In another embodiment, the present disclosure provides an anti-RSV antibody composition consisting of: a) an anti-RSV antibody or antigen binding fragment thereof at a concentration of about 100 mg/ml; b) histidine at a concentration of about 10 mM; c) sucrose at a concentration of about 7% in a mass-volume ratio (w/v); d) a mixture of L-arginine at a concentration of 25 mM (HCl) and L-lysine (HCl) at a concentration of 25 mM; e) DTPA at a concentration of 50 µM and f) polysorbate 80 at a concentration of 0.02%.

In one aspect, the present disclosure provides an anti-RSV antibody composition comprising: a) an anti-RSV antibody or antigen binding fragment thereof at a concentration of about 50 mg/ml to about 250 mg/ml; b) histidine at a concentration of about 5 mM to about 20 mM; c) sucrose at a concentration of about 4% to about 8% w/v; d) an excipient selected from the group consisting of the following: L-arginine or a pharmaceutically acceptable L-arginine salt at a concentration of about 25 mM to about 75 mM, L-lysine or a pharmaceutically acceptable L-lysine salt at a concentration of about 25 mM to about 75 mM, a mixture of L-arginine or a pharmaceutically acceptable salt of L-arginine at a concentration of about 25 mM to about 75 mM and L-glutamate or a pharmaceutically acceptable salt of L-glutamate at a concentration of about 25 mM to about 75 mM and a mixture of L- arginine or a pharmaceutically acceptable L-arginine salt at a concentration of about 25 mM to about 75 mM and L-lysine or a pharmaceutically acceptable L-lysine salt at a concentration from about 25 mM to about 75 mM and e) polysorbate 80 at a concentration of about 0. 01% to about 0.10% w/v.

In one embodiment, the anti-RSV antibody composition comprises: a) an anti-RSV antibody or antigen-binding fragment thereof at a concentration of about 50 mg/ml to about 250 mg/ml; b) histidine at a concentration of about 5 mM to about 20 mM; c) sucrose at a concentration of about 4% to about 8% w/v; d) L-arginine or a pharmaceutically acceptable L-arginine salt at a concentration of about 25 mM to about 75 mM; and e) polysorbate 80 at a concentration of about 0.01% to about 0.10% w/v. In one embodiment, the anti-RSV antibody composition comprises: a) an anti-RSV antibody or antigen-binding fragment thereof at a concentration of about 50 mg/ml to 250 mg/ml; b) histidine at a concentration of 5 mM to 20 mM; c) sucrose at a concentration of 4% to 8% w/v; d) L-arginine or a pharmaceutically acceptable L-arginine salt at a concentration of 25 mM to 75 mM; and e) polysorbate 80 at a concentration of 0.01% to 0.10% w/v.

In one embodiment, the anti-RSV antibody composition comprises: a) an anti-RSV antibody or antigen-binding fragment thereof at a concentration of about 50 mg/ml to about 250 mg/ml; b) histidine at a concentration of about 5 mM to about 20 mM; c) sucrose at a concentration of about 4% to about 8% w/v; d) L-lysine or a pharmaceutically acceptable L-lysine salt at a concentration of about 25 mM to about 75 mM; and e) polysorbate 80 at a concentration of about 0.01% to about 0.10% w/v. In one embodiment, the anti-RSV antibody composition comprises: a) an anti-RSV antibody or antigen-binding fragment thereof at a concentration of about 50 mg/ml to 250 mg/ml; b) histidine at a concentration of 5 mM to 20 mM; c) sucrose at a concentration of 4% to 8% w/v; d) L-lysine or a pharmaceutically acceptable L-lysine salt at a concentration of 25 mM to 75 mM; and e) polysorbate 80 at a concentration of 0.01% to 0.10% w/v.

In one embodiment, the anti-RSV antibody composition comprises: a) an anti-RSV antibody or antigen-binding fragment thereof at a concentration of about 50 mg/ml to about 250 mg/ml; b) histidine at a concentration of about 5 mM to about 20 mM; c) sucrose at a concentration of about 4% to about 8% w/v; d) a mixture of L-arginine or a pharmaceutically acceptable salt of L-arginine at a concentration of about 25 mM to about 75 mM and L-glutamate or a pharmaceutically acceptable salt of L-glutamate at a concentration of about 25 mM to about 75 mM and e) polysorbate 80 b concentrations from about 0.01% to about 0.10% w/v. In one embodiment, the anti-RSV antibody composition comprises: a) an anti-RSV antibody or antigen-binding fragment at a concentration of 50 mg/ml to 250 mg/ml; b) histidine at a concentration of 5 mM to 20 mM; c) sucrose at a concentration of 4% to about 8% w/v; d) a mixture of L-arginine or a pharmaceutically acceptable salt of L-arginine at a concentration of 25 mM to 75 mM and L-glutamate or a pharmaceutically acceptable salt of L-glutamate at a concentration of 25 mM to 75 mM and e) polysorbate 80 at a concentration of 0. 01% to 0.10% w/v.

In one embodiment, the anti-RSV antibody composition comprises: a) an anti-RSV antibody or antigen-binding fragment thereof at a concentration of about 50 mg/ml to about 250 mg/ml; b) histidine at a concentration of about 5 mM to about 20 mM; c) sucrose at a concentration of about 4% to about 8% w/v; d) a mixture of L-arginine or a pharmaceutically acceptable salt of L-arginine at a concentration of about 25 mM to about 75 mM and L-lysine or a pharmaceutically acceptable salt of L-lysine at a concentration of about 25 mM to about 75 mM and e) polysorbate 80 b concentrations from about 0.01% to about 0.10% w/v. In one embodiment, the anti-RSV antibody composition comprises: a) an anti-RSV antibody or antigen-binding fragment thereof at a concentration of about 50 mg/ml to 250 mg/ml; b) histidine at a concentration of 5 mM to 20 mM; c) sucrose at a concentration of 4% to 8% w/v; d) a mixture of L-arginine or a pharmaceutically acceptable salt of L-arginine at a concentration of 25 mM to 75 mM and L-lysine or a pharmaceutically acceptable salt of L-lysine at a concentration of 25 mM to 75 mM and e) polysorbate 80 at a concentration of 0. 01% to 0.10% w/v.

In some embodiments, the composition has a pH value between 5.5 and 6.5.

In some embodiments, the composition has a pH value of 6.0.

Also provided herein are methods of preventing and/or treating RSV-related diseases in a human patient in need thereof, comprising: administering to the patient an effective amount of the kRSV antibody formulations of the present invention.

Also provided herein are methods of treating or preventing RSV infection in a human patient in need thereof, comprising: administering to the patient an effective amount of the anti-RSV antibody compositions of the present invention.

Also provided herein are methods of treating or preventing RSV infection in a human patient in need thereof, comprising: administering to the patient an effective amount of an anti-RSV antibody composition of the present invention, wherein the effective amount includes a fixed dose of an anti-RSV antibody or an antigen binding fragment thereof, wherein the fixed dose ranges from about 10 to about 100 mg.

Also provided herein are methods of treating or preventing RSV infection in a human patient in need thereof, comprising: administering to the patient an effective amount of the anti-RSV antibody formulations of the present invention, wherein the anti-RSV antibody formulations are administered subcutaneously or wherein the anti-RSV antibody formulations are administered intravenously; or wherein the anti-RSV antibody compositions are administered intramuscularly.

Also provided herein is the use of the anti-RSV antibody compositions of the present invention for the treatment or prevention of RSV infection in a human patient.

Brief description of drawings

In fig. 1A-1C show the UPSEC results of %HMW, %mAb (monomer) and %LMW as a function of time for RSV antibody formulations at 5°C storage conditions.

In fig. 2A-2C show the UPSEC results: %HMW, %mAb (monomer) and %LMW as a function of time for RSV antibody formulations under 25°C storage conditions.

In fig. 3A-3C show the UPSEC results: %HMW, %mAb (monomer) and %LMW as a function of time for RSV antibody formulations under storage conditions at 40°C.

In fig. 4A-4B show the HP-IEX results of % total acid peaks and % analyte peak versus time for anti-RSV antibody formulations at 5°C storage conditions.

In fig. 5A-5B show the HP-IEX results of % total acid peaks and % analyte peak versus time for anti-RSV antibody formulations at 25°C storage conditions.

In fig. 6A-6B show HP-IEX results of % total acid peaks and % analyte peak as a function of time for anti-RSV antibody formulations under storage conditions at 40°C.

In fig. Figure 7 shows the results of the viscosity reduction of the anti-RSV antibody excipient at a concentration of 100 mg/ml (all experiments were repeated n=5 to obtain the standard deviation).

In fig. 8A-8C show the UPSEC results of %HMW, %mAb (monomer) and %LMW as a function of time for RSV antibody formulations at 5°C storage conditions.

In fig. 9A-9C show the UPSEC results of %HMW, %mAb (monomer) and %LMW as a function of time for RSV antibody formulations at 25°C storage conditions.

In fig. 10A-10C show the UPSEC results: %HMW, %mAb (monomer) and %LMW as a function of time for RSV antibody formulations under storage conditions at 40°C.

In fig. 11A-11B show HP-IEX results of % total acid peaks and % analyte peak as a function of time for anti-RSV antibody formulations at 5°C storage conditions.

In fig. 12A-12B show HP-IEX results of % total acid peaks and % analyte peak as a function of time for anti-RSV antibody formulations at 25°C storage conditions.

In fig. 13A-13B show the HP-IEX results of % total acid peaks and % analyte peak as a function of time for anti-RSV antibody formulations under storage conditions at 40°C.

In fig. Figure 14 shows the percentage degradation results of PS80 at 40°C.

In fig. Figure 15 shows a bar graph comparing the viscosity of formulations with excipients of 80 mM histidine, 70 mM lysine, or 70 mM arginine.

Detailed Description of the Present Invention

The present invention provides stable formulations containing an anti-RSV antibody or an antigen-binding fragment thereof that binds to RSV. The present invention provides stable formulations containing an anti-RSV antibody or an antigen-binding fragment thereof that binds to the human RSV F protein, which are useful for methods of treating or preventing human RSV infection.

Antibodies used in the compositions disclosed herein are described in US Pat. No. 9,963,500. An exemplary antibody used in the compositions disclosed herein is described in US Pat. No. 9,963,500. This exemplary antibody contains complementarity determining regions (CDRs) having the amino acid sequences: SEQ ID NO: 1 (heavy chain CDR 1), SEQ ID NO: 2 (heavy chain CDR 2), SEQ ID NO: 3 (heavy chain CDR 3) , SEQ ID NO: 4 (light chain CDR 1), SEQ ID NO: 5 (light chain CDR 2) and SEQ ID NO: 6 (light chain CDR 3). In addition, this exemplary antibody comprises heavy and light chain variable regions having the amino acid sequences of SEQ ID NO: 7 and SEQ ID NO: 8. In addition, this exemplary antibody includes immunoglobulin heavy and light chains consisting of the amino acid sequences of SEQ ID NO: 9 and SEQ ID NO: 10.

I. Definitions and abbreviations

Definitions are used throughout the description. Abbreviations are defined throughout the specification and claims.

For easier understanding of the present invention, certain technical and scientific terms are specifically defined below. Unless specifically defined elsewhere in this document, all other technical and scientific terms used herein have the meaning commonly understood by one skilled in the art to which the present invention belongs.

As used throughout the specification and the accompanying claims, the singular number includes the plural unless the context clearly dictates otherwise.

A reference to "or" indicates one or both possibilities unless the context clearly dictates one of the specified possibilities. In some cases, “and/or” was used to emphasize one or both possibilities.

When a range of values is given, such as "an amount from 50 mg to 100 mg", the range is intended to include the values stated.

"Administration" and "exposure" when applied to an animal, human, experimental subject, cell, tissue, organ or biological fluid refer to the contact of an exogenous pharmaceutical, therapeutic, diagnostic agent or composition with the animal, human, subject, cell, tissue, organ or biological fluid liquid. The effect on cells involves contact of the reagent with the cell, as well as contact of the reagent with the liquid, and the liquid is in contact with the cell. “Administration” and “exposure” also mean exposure in vitro and ex vivo, for example, to cells, a reagent, a diagnostic compound, a binding compound, or another cell.

“RSV-associated disease” means any disease caused, directly or indirectly, by respiratory syncytial virus (RSV) infection, as well as diseases or conditions that predispose a patient to RSV infection. Examples of diseases that fall into the first category include pneumonia and bronchiolitis. Diseases and conditions that fall into the latter category (i.e., those that place the patient at risk for severe RSV infection) include cystic fibrosis, congenital heart disease, cancer, age-related immunosuppression, transplant recipients, and generally any condition that causes the condition immunosuppression. or decreased immune system function, such as postoperative organ transplant regimens or preterm birth.

“Treat” or “treatment” means the administration of a therapeutic agent, such as a composition containing any of the antibodies or antigen-binding fragments of the present invention, internally or externally to a subject or patient having one or more symptoms of a disease, or suspected of having a disease for which the agent has therapeutic activity. Typically, the agent is administered in an amount effective to relieve one or more symptoms of a disease in a subject or population being treated, either by inducing regression or inhibiting the progression of such symptom(s) to any clinically measurable extent. The amount of therapeutic agent that is effective in alleviating any particular symptom of a disease may vary depending on factors such as the disease state, the age and weight of the patient, and the ability of the drug to produce the desired response in the subject. Whether a disease symptom has improved can be assessed using any clinical measurement typically used by physicians or other qualified health care providers to assess the severity or progression status of that symptom. RSV antibody treatment can also be combined with other interventions (antibodies, nucleic acids, vaccines, and small molecules) to treat other respiratory pathogens.

“Prevent” or “prophylaxis” means to administer a prophylactic agent, such as a composition containing any of the antibodies or antigen-binding fragments of the present invention, internally or externally to a subject or patient at risk for RSV infection for which the agent has prophylactic activity. Prevention includes reducing the likelihood or severity of subsequent RSV infection, alleviating symptoms associated with lower respiratory tract infection (LRI) following RSV infection, and inducing immunity to protect against RSV infection. Typically, the agent is administered in an amount effective to neutralize RSV in the lungs and/or nose to block infection. The amount of prophylactic agent that is effective in alleviating the symptoms of any particular disease may vary depending on factors such as the age and weight of the patient, as well as the ability of the agent to produce the desired response in the subject. Whether a disease symptom has improved can be assessed using any clinical measurement typically used by physicians or other qualified health care providers to assess the severity or progression status of that symptom or, in some cases, the reduction in the need for hospitalization.

The term “patient” (alternatively referred to herein as “subject” or “individual”) refers to a mammal (eg, rat, mouse, dog, cat, rabbit) capable of being treated with the compositions of the present invention, most preferably a human. In some embodiments, the patient is an adult patient. In other embodiments, the patient is a child patient. Those "in need of treatment" include those patients who may benefit from treatment with the compositions of the present invention.

The term "pharmaceutically effective amount" or "effective amount" means the amount at which a therapeutic composition is administered to a patient to treat a disease or condition. One skilled in the art will recognize that this level may vary depending on patient characteristics such as age, weight, etc.

As used herein, the phrase “fixed dose” refers to the amount (eg, in milligrams) of the active ingredient that is administered to the patient.

The term “about” when changing the amount (e.g., mM or M) of a substance or composition, the percentage (volume or mass-volume ratio) of a component of a composition, the pH of a solution/formulation, or the value of a parameter characterizing a process step, or the like. , refers to a change in a numerical value by plus or minus 5%. Such change in numerical quantity may occur, for example, through typical measurement, handling and sampling procedures associated with the preparation, characterization and/or use of the substance or composition; due to an unintentional error in these procedures; due to differences in the manufacture, source or purity of ingredients used to make or use the compositions or perform the procedures; etc.

The phrase "consisting essentially of" or variations such as "consisting essentially of" or "consisting essentially of" used in the specification and claims indicate the inclusion of any listed elements or group of elements and the optional inclusion of other elements similar or different in nature from the listed elements, which do not significantly change the basic or new properties of the specified administration regimen, method or composition. As a non-limiting example, a binding compound that consists essentially of the specified amino acid sequence may also include one or more amino acids, including substitutions of one or more amino acid residues that do not significantly affect the properties of the binding compound.

“Comprising” or variants such as “comprise”, “comprises” or “consists of” are used throughout the specification and claims in the inclusive sense, that is, to indicate the presence of the claimed features, but not to exclude the presence or add additional functions, which may substantially improve the operation or usefulness of any embodiment of the present invention unless the context otherwise requires due to language or necessary meaning.

The phrase "pharmaceutical formulation" refers to preparations that are in such a form as to allow the active ingredients to be effective and that do not contain additional components that are toxic to subjects to whom the formulation may be administered. The terms "composition" and "pharmaceutical composition" are used interchangeably throughout.

The phrase "pharmaceutically acceptable" refers to excipients (carriers, additives) and compositions that can be reasonably administered to a subject to provide an effective dose of the active ingredient administered and that are "generally considered safe", e.g., that are physiologically tolerable and generally do not cause an allergic or similar undesirable reaction such as stomach upset and the like when administered to a human. In another embodiment, the term refers to molecular entities and compositions approved by a federal or state government regulatory agency or listed in the United States Pharmacopoeia or other generally accepted pharmacopoeia for use in animals and, in particular, in humans.

As used herein, the term “effective amount” means that amount of an active compound or pharmaceutical agent that produces a biological or therapeutic response in a tissue, system, animal or human as desired by a researcher, veterinarian, physician or other clinician. In one embodiment, the effective amount is a "therapeutically effective amount" for alleviating the symptoms of the disease or condition being treated. In another embodiment, the effective amount is a “prophylactically effective amount” for preventing symptoms of a preventable disease or condition.

A "stable" formulation is one in which the protein therein substantially retains its physical stability and/or chemical stability and/or biological activity upon storage. Various analytical methods for measuring protein stability are available in the art and are discussed in Peptide and Protein Drug Delivery, 247-301, Vincent Lee Ed., Marcel Dekker, Inc., New York, N.Y., Pubs. (1991) and Jones, A. Adv. Drug Delivery Rev. 10:29-90 (1993). Stability can be measured at a selected temperature for a selected period of time. For example, in one embodiment, a stable formulation is one that does not exhibit significant changes at refrigeration temperature (2-8°C) for at least 12 months. According to another embodiment, a stable formulation is one that does not exhibit significant changes at refrigeration temperature (2-8°C) for at least 18 months. According to another embodiment, a stable formulation is one that does not exhibit significant changes at room temperature (23-27°C) for at least 3 months. According to another embodiment, a stable formulation is one that does not exhibit significant changes at room temperature (23-27°C) for at least 6 months. According to another embodiment, a stable formulation is one that does not exhibit significant changes at room temperature (23-27°C) for at least 12 months. In another embodiment, a stable formulation is one that does not exhibit significant changes at room temperature (23-27°C) for at least 18 months. The stability criteria for the antibody composition are as follows. Typically, no more than 10%, preferably 5%, of the antibody monomer is degraded as measured by SEC-HPLC. Typically, the composition is colorless or transparent or slightly opalescent when visually analyzed. Typically, the concentration, pH and osmolality of the formulation will vary by no more than +/- 10%. Efficiency is typically in the range of 60-140%, preferably 80-120%, of the control or comparison. Typically, no more than 10%, preferably 5%, antibody cutoff is observed, i.e. % low molecular weight particles as determined, for example, by HP-SEC. Typically, no more than 10%, preferably no more than 5%, of antibody aggregation is observed, i.e. % high molecular weight particles as determined, for example, by HP-SEC.

The phrase "anti-RSV antibody" refers to a monoclonal antibody directed against the F protein or the G protein of RSV. Antibodies to RSV are disclosed and described in US Pat. No. 9,963,500. A specific anti-RSV antibody is disclosed and described in US Pat. No. 9,963,500 and contains complementarity determining regions (CDRs) having the amino acid sequences: SEQ ID NO: 1 (heavy chain CDR 1), SEQ ID NO: 2 (heavy chain CDR 2), SEQ ID NO: 3 (heavy chain CDR 3), SEQ ID NO: 4 (light chain CDR 1), SEQ ID NO: 5 (light chain CDR 2) and SEQ ID NO: 6 (light chain CDR 3). In addition, an anti-RSV antibody is disclosed and described in US Pat. No. 9,963,500 and contains heavy and light chain variable regions having the amino acid sequences of SEQ ID NO: 7 and SEQ ID NO: 8, respectively. In addition, a specific anti-RSV antibody is disclosed and described in US Pat. No. 9,963,500 and includes immunoglobulin heavy and light chains consisting of the amino acid sequences SEQ ID NO: 9 and SEQ ID NO: 10, respectively.

An antibody "maintains its physical stability" in a pharmaceutical formulation if it does not show a significant increase in aggregation, precipitation and/or denaturation when examined visually for color and/or clarity or when measured by UV light scattering, size exclusion chromatography (SEC) and dynamic scattering Sveta. Changes in protein conformation can be assessed by fluorescence spectroscopy, which determines the tertiary structure of the protein, and by FTIR spectroscopy, which determines the secondary structure of the protein.

An antibody "retains its chemical stability" in a pharmaceutical formulation unless it exhibits significant chemical changes. Chemical stability can be assessed by detecting and quantifying chemically altered forms of the protein. Degradation processes that often change the chemical structure of a protein include hydrolysis or truncation (assessed by techniques such as size exclusion chromatography and SDS-PAGE), oxidation (assessed by techniques such as peptide mapping coupled with mass spectroscopy or MALDI/TOF). MS), deamidation (assessed by methods such as ion exchange chromatography, capillary isoelectric focusing, peptide mapping, isoaspartic acid measurement) and isomerization (assessed by isoaspartic acid measurement, peptide mapping, etc.).

An antibody "retains its biological activity" in a pharmaceutical composition if the biological activity of the antibody at a given time is within the predetermined range of biological activity exhibited during the preparation of the pharmaceutical composition. The biological activity of an antibody can be determined, for example, using an antigen binding assay. The compositions of the present invention include antibodies and fragments thereof that are biologically active in reconstituted or liquid form.

II. Compositions of the present invention

The compositions of the present invention minimize the formation of antibody aggregates, increase stability and reduce viscosity.

The present invention includes various formulations of an anti-RSV antibody or antigen-binding fragment thereof, as described in more detail below. For example, the present invention includes compositions containing (i) an anti-RSV antibody or antigen-binding fragment thereof, (ii) a buffer (eg, histidine), (iii) a stabilizer (eg, a non-reducing sugar such as sucrose); (iv) a nonionic surfactant (eg, polysorbate 80); and (v) an antioxidant (eg, methionine). In further embodiments, the compositions of the present invention comprise a viscosity-lowering agent (e.g., arginine or a pharmaceutically acceptable salt thereof; lysine or a pharmaceutically acceptable salt thereof; a mixture of arginine and lysine or pharmaceutically acceptable salts thereof, a mixture of arginine and glutamate or pharmaceutically acceptable salts thereof, and /or histidine or pharmaceutically acceptable salts thereof) and/or a metal chelator (eg DTPA).

III. Antibodies to RSV and their antigen-binding fragments

The present invention provides stable biological compositions containing an anti-RSV antibody that contains complementarity determining regions (CDRs) having the amino acid sequences: SEQ ID NO: 1 (heavy chain CDR 1), SEQ ID NO: 2 (heavy chain CDR 2), SEQ ID NO: 3 (heavy chain CDR 3), SEQ ID NO: 4 (light chain CDR 1), SEQ ID NO: 5 (light chain CDR 2) and SEQ ID NO: 6 (light chain CDR 3). The present invention also provides stable biological compositions containing an anti-RSV antibody that contains heavy and light chain variable regions having the amino acid sequences of SEQ ID NO: 7 and SEQ ID NO: 8, respectively, and/or containing immunoglobulin heavy and light chains, consisting of the amino acid sequences SEQ ID NO: 9 and SEQ ID NO: 10, respectively. Methods for producing anti-RSV antibodies are disclosed and described in US Pat. No. 9,963,500 and are incorporated herein by reference in their entirety. SEQ ID NO: 1-10 are presented in Table A below:

In some embodiments of the formulations described herein, the API (“active pharmaceutical ingredient,” i.e., an anti-RSV antibody or antigen-binding fragment thereof) is present at a concentration of about 50 mg/mL to about 250 mg/mL. In some embodiments of the formulations, the API (ie, an anti-RSV antibody or antigen-binding fragment thereof) is present at a concentration of from about 75 mg/ml to about 200 mg/ml. In some embodiments, the API is present at a concentration of from about 90 mg/ml to about 110 mg/ml. In another embodiment, the API is present at a concentration of about 100 mg/ml. In another embodiment, the API is present at a concentration of 90 mg/ml to 110 mg/ml. In another embodiment, the API is present at a concentration of 100 mg/ml. In another embodiment, the API is present at a concentration of 75 mg/ml. In another embodiment, the API is present at a concentration of 50 mg/ml.

In some embodiments, the API is present at a concentration of from about 125 mg/ml to about 175 mg/ml. In some embodiments, the API is present at a concentration of from about 140 mg/ml to about 160 mg/ml. In another embodiment, the API is present at a concentration of about 150 mg/ml. In some embodiments, the API is present at a concentration of 125 mg/ml to 175 mg/ml. In some embodiments, the API is present at a concentration of 140 mg/ml to 160 mg/ml. In another embodiment, the API is present at a concentration of 150 mg/ml.

In some embodiments, the API is present at a concentration of from about 150 mg/ml to about 200 mg/ml. In some embodiments, the API is present at a concentration of from about 165 mg/ml to about 185 mg/ml. In another embodiment, the API is present at a concentration of about 175 mg/ml. In some embodiments, the API is present at a concentration of 150 mg/ml to 200 mg/ml. In some embodiments, the API is present at a concentration of 165 mg/ml to 185 mg/ml. In another embodiment, the API is present at a concentration of 175 mg/ml.

In some embodiments, the API is present at a concentration of from about 175 mg/ml to about 225 mg/ml. In some embodiments, the API is present at a concentration of from about 190 mg/ml to about 210 mg/ml. In another embodiment, the API is present at a concentration of about 200 mg/ml. In some embodiments, the API is present at a concentration of 175 mg/ml to 225 mg/ml. In some embodiments, the API is present at a concentration of 190 mg/ml to 210 mg/ml. In another embodiment, the API is present at a concentration of 200 mg/ml.

In some embodiments, the API is present at a concentration of from about 200 mg/ml to about 250 mg/ml. In some embodiments, the API is present at a concentration of from about 215 mg/ml to about 235 mg/ml. In another embodiment, the API is present at a concentration of about 225 mg/ml. In some embodiments, the API is present at a concentration of 200 mg/ml to 250 mg/ml. In some embodiments, the API is present at a concentration of 215 mg/ml to 235 mg/ml. In another embodiment, the API is present at a concentration of 225 mg/ml.

In some embodiments of the formulations disclosed herein, the API (ie, an anti-RSV antibody or antigen-binding fragment thereof) is present in a fixed dose (eg, a milligram amount). In some embodiments of the formulations disclosed herein, the API is present at a fixed dose of about 10 mg to about 150 mg. In some embodiments, the API is present at a fixed dose of 10 mg to 150 mg. In some embodiments, the API is present at a fixed dose of about 25 mg to about 125 mg. In some embodiments, the API is present at a fixed dose of 25 mg to 125 mg. In some embodiments, the API is present at a fixed dose of about 50 mg to about 100 mg. In some embodiments, the API is present at a fixed dose of 50 mg to 100 mg.

IV. Excipients of the composition

The compositions of the present invention contain at least one excipient that stabilizes the composition.

In some embodiments of the compositions of the present invention, the stabilizer is a non-reducing sugar. In embodiments of the present invention, the non-reducing sugar is sucrose. In further embodiments, the non-reducing sugar is glucose. In further embodiments, the non-reducing sugar is trehalose. In other embodiments, the non-reducing sugar is lactose. In other embodiments, the non-reducing sugar is raffinose.

In some embodiments, the anti-RSV antibody compositions of the present invention comprise a stabilizer selected from the group consisting of: sucrose, glucose, trehalose, lactose, or raffinose at a concentration of from about 4% to about 8% on a mass-volume (w/v) basis. .

In some embodiments, the stabilizer is sucrose at a concentration of from about 4% to about 8% on a weight-volume basis. In some embodiments, the stabilizer is sucrose at a concentration of 4% to 8% w/v.

In some embodiments, the stabilizer is sucrose at a concentration of from about 6% to about 8% on a weight-volume basis.

In some embodiments, the stabilizer is sucrose at a concentration of 4% to 8% w/v.

In some embodiments, the stabilizer is trehalose at a concentration of about 4% to about 8% w/v. In some embodiments, the stabilizer is trehalose at a concentration of 4% to 8% w/v.

In some embodiments, the stabilizer is trehalose at a concentration of about 6% to about 8% w/v. In some embodiments, the stabilizer is trehalose at a concentration of 6% to 8% w/v.

In some embodiments, the stabilizer is sucrose at a concentration of from about 6.5% to about 7.5% on a weight-volume basis. In some embodiments, the stabilizer is sucrose at a concentration of 6.5% to 7.5% w/v.

In some embodiments, the stabilizer is sucrose at a concentration of 6% to 8%.

In some embodiments, the stabilizer is sucrose at a concentration of 6.5% to 7.5% w/v.

In some embodiments, the stabilizer is sucrose at a concentration of 7%.

The compositions of the present invention contain arginine, for example L-arginine, or a pharmaceutically acceptable salt thereof, for example HCl; lysine, eg L-lysine, or a pharmaceutically acceptable salt thereof, eg HCl; or a combination of arginine (L-arginine) or a pharmaceutically acceptable salt thereof (HCl) and lysine (L-lysine) or a pharmaceutically acceptable salt thereof (HCl); or a combination of arginine (L-arginine) or a pharmaceutically acceptable salt thereof (HCl) and glutamate (L-glutamate) or a pharmaceutically acceptable salt thereof; all this can provide additional stability to the composition, as well as regulate viscosity, which allows the creation of compositions with high concentrations of API.

In addition to the anti-RSV antibody or antigen-binding fragment thereof and a stabilizer in the amounts/concentrations specified above, the compositions of the present invention also contain a buffer. In some embodiments, the buffer is present in an amount from about 5 mM to about 20 mM, which provides a pH value in the range of about 5 to about 7. In some embodiments, the buffer is present in an amount from 5 mM to 20 mM, which provides a pH value in the range of range from 5 to 7.

In some embodiments of the present invention, the buffer provides the pH of the formulation in the range of about 5.5 to about 6.5. According to some embodiments of the invention, the buffer provides a pH value of the composition in the range from 5.5 to 6.5. In some embodiments, the buffer has a pH value in the range of about 6.0. In other embodiments, the pH value is 6.0.

In certain embodiments, the buffer has a pH value of about 5.7, about 5.8, about 5.9, about 6.0, about 6.1, about 6.2, or about 6.3. In certain embodiments, the buffer has a pH value of 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, or 6.3. Examples of buffers that will control pH in this range include succinate (sodium or potassium), histidine, sodium acetate, phosphate (sodium or potassium), tris (tris(hydroxymethyl)aminomethane), diethanolamine, citrate (sodium), and other organic acid buffers.

In some embodiments of the present invention, the buffer is a histidine or acetate with a pH value of from about 5.5 to about 6.5. In some embodiments of the present invention, the buffer is histidine or acetate with a pH value between 5.5 and 6.5. In some embodiments, the buffer is an L-histidine buffer. In embodiments where the formulation is lyophilized, it is preferred that the buffer is non-acetate since acetate buffer systems are incompatible with the lyophilization process.

When a range of pH values is specified, such as “pH 5.5 to 6.5,” the range is intended to include the stated values. Unless otherwise stated, for a lyophilized formula, pH refers to the pH after reconstitution of the lyophilized formulations of the present invention. pH is typically measured at 25°C using a standard pH meter with a glass bulb. As used herein, a solution containing “histidine buffer at pH X” refers to a solution at pH X and containing a histidine buffer, i.e. pH is intended to indicate the pH of a solution.

In addition to the anti-RSV antibody or antigen-binding fragment thereof, stabilizer and buffer in the amounts/concentrations specified above, the compositions of the present invention also contain an antioxidant. In embodiments of the present invention, the antioxidant is methionine. In embodiments of the present invention, the antioxidant is L-methionine or a pharmaceutically acceptable salt thereof. In further embodiments, the methionine is L-methionine. In other embodiments, the antioxidant is L-methionine HCl. In other embodiments, the antioxidant is histidine.

In some embodiments, the antioxidant (eg, L-methionine) is present in the compositions of the present invention in an amount of from about 1 mM to about 20 mM. In some embodiments, the antioxidant is present in an amount of about 5 mM to about 20 mM, about 5 mM to about 15 mM, or about 5 mM to about 10 mM. In some embodiments, the antioxidant is present in an amount of about 1 mM, about 2 mM, about 3 mM, about 4 mM, about 5 mM, about 6 mM, about 7 mM, about 8 mM, about 9 mM, about 10 mM, about 11 mM, about 12 mM, about 13 mM, about 14 mM, about 15 mM, about 16 mM, about 17 mM, about 18 mM, about 19 mM, or about 20 mM. In some embodiments, the antioxidant is present in an amount of 5 to 20 mM, 5 to 15 mM, or 5 to 10 mM. In some embodiments, the antioxidant is present in an amount of 1 mM, 2 mM, 3 mM, 4 mM, 5 mM, 6 mM, 7 mM, 8 mM, 9 mM, 10 mM, 11 mM, 12 mM, 13 mM, 14 mM, 15 mM, 16 mM, 17 mM, 18 mM, 19 mM or 20 mM.

In embodiments where the antioxidant is histidine, histidine may be present in amounts up to 100 mM. In such embodiments, histidine can serve as a buffer, antioxidant, and/or reduce the viscosity of the formulations described herein. In some embodiments, histidine may be present at a concentration of about 10-20 mM, about 20-30 mM, about 30-40 mM, about 40-50 mM, about 50-60 mM, about 60-70 mM, about 70 mM-80 mM, about 80-90 mM or about 90-100 mM. In some embodiments, histidine may be present at concentrations of 10-20 mM, 20-30 mM, 30-40 mM, 40-50 mM, 50-60 mM, 60-70 mM, 70-80 mM, 80-90 mM, or 90 -100 mM. In some embodiments, histidine may be present at a concentration of about 10 mM, about 20 mM, about 30 mM, about 40 mM, about 50 mM, about 60 mM, about 70 mM, about 80 mM, about 90 mM, or about 100 mM. In some embodiments, histidine may be present at a concentration of 10 mM, 15 mM, 20 mM, 25 mM, 30 mM, 35 mM, 40 mM, 45 mM, 50 mM, 55 mM, 60 mM, 65 mM, 70 mM, 75 mM , 80 mM, 85 mM, 90 mM, 95 mM or 100 mM.

In addition to the anti-RSV antibody or antigen-binding fragment thereof, stabilizer, buffer and antioxidant in the amounts/concentrations specified above, the compositions of the present invention also contain a surfactant. Surfactants are typically added to formulations to provide stability, reduce and/or prevent aggregation, or to prevent and/or inhibit protein damage during processing conditions such as purification, filtration, freeze-drying, transportation, storage and delivery. In some embodiments of the present invention, the surfactant is useful to provide additional stability to the active ingredient(s), i.e. antibodies to RSV or its antigen-binding fragment.

Surfactants that can be used in the compositions of the present invention include, but are not limited to: nonionic surfactants such as polyoxyethylene sorbitan fatty acid esters (polysorbates, sold under the trade name TWEEN (Uniquema Americas LLC, Wilmington , Delaware)), including polysorbate-20 (polyoxyethylene sorbitan monolaurate), polysorbate-40 (polyoxyethylene sorbitan monopalmitate), polysorbate-60 (polyoxyethylene sorbitan monostearate) and polysorbate-80 (polyoxyethylene sorbitan monooleate); polyoxyethylene alkyl ethers such as BRIJ 58 (Uniquema Americas LLC, Wilmington, DE) and BRIJ 35; poloxamers (eg poloxamer 188); TRITON X-100 (Union Carbide Corp., Houston, TX) and TRITON X-114; NP40; Span 20, Span 40, Span 60, Span 65, Span 80 and Span 85; copolymers of ethylene and propylene glycol (for example, the PLURONIC series of nonionic surfactants such as PLURONIC F68, PLURONIC 10R5, PLURONIC F108, PLURONIC F127, PLURONIC F38, PLURONIC L44, PLURONIC L62 (BASF Corp., Ludwigshafen, Germany) and dodecyl sulf sodium at ( DSN).

The amount of surfactant to be included in the compositions of the present invention is an amount sufficient to perform the desired function, i.e. the minimum amount required to stabilize the active pharmaceutical ingredient (i.e., anti-RSV antibody or antigen-binding fragment thereof) in the formulation. Typically, the surfactant is present in a concentration of from about 0.008% to about 0.1% on a weight-volume basis. In some embodiments of this aspect of the present invention, the surfactant is present in the composition in an amount from about 0.01% to about 0.04% w/v; from about 0.01% to about 0.03% w/v, from about 0.01% to about 0.02% w/v, from about 0.015% to about 0.04% w/v attitude; from about 0.015% to about 0.03% w/v, from about 0.015% to about 0.02% w/v, from about 0.02% to about 0.04% w/v, from about 0.02% to about 0.035% on a weight-volume basis, or from about 0.02% to about 0.03% on a weight-volume basis. In particular embodiments, the surfactant is present in an amount of about 0.02% on a weight-volume basis. mass-volume ratio. In alternative embodiments, the surfactant is present in an amount of about 0.01%, about 0.015%, about 0.025%, about 0.03%, about 0.035%, or about 0.04% on a weight-volume basis.

In exemplary embodiments of the present invention, the surfactant is a nonionic surfactant selected from the group consisting of polysorbate 20, polysorbate 80, and F127. In preferred embodiments, the surfactant is polysorbate 80.

In certain embodiments, anti-RSV antibody formulations contain PS80 in an amount of from about 0.01% to about 0.04% on a weight-volume basis. In further embodiments, anti-RSV antibody formulations contain PS80 in an amount of about 0.008%, about 0.01%, about 0.015%, about 0.02%, about 0.025%, about 0.03%, about 0.035%, about 0.04 % or about 0.045% in mass-volume ratio. In certain embodiments, anti-RSV antibody formulations contain PS80 in an amount of about 0.02% w/v. In some embodiments, anti-RSV antibody formulations contain PS80 in an amount of 0.01% to 0.04% w/v. In some embodiments, anti-RSV antibody formulations contain PS80 at an amount of 0.008%, 0.01%, 0.015%, 0.02%, 0.025%, 0.03%, 0.035%, 0.04%, or 0.045% w/v In certain embodiments, anti-RSV antibody formulations contain PS80 at 0.02% w/v.

In certain embodiments, the anti-RSV antibody compositions of the present invention contain a chelator selected from DTPA and EDTA. In some embodiments, the chelator is DTPA. In further embodiments, the amount of DTPA is from about 10 μM to about 90 μM, from about 25 μM to about 75 μM, or about 50 μM. In some embodiments, the chelator is DTPA. In some embodiments, the amount of DTPA is 10 μM to 90 μM, 25 μM to 75 μM, or 50 μM.

In addition to the anti-RSV antibody or antigen-binding fragment thereof, embodiments of the formulation may contain a stabilizer, buffer, antioxidant, surfactant, and chelator in the amounts/concentrations specified above.

The present disclosure also provides an anti-RSV antibody composition as described herein, the composition being contained in a glass vial or injection device (eg, syringe).

In additional embodiments, the present disclosure provides anti-RSV antibody formulations described herein wherein, after storing the formulation at 2-8°C for 12 months, the percentage of heavy chain and light chain measured by CE-SDS under reducing conditions is > 96%.

In additional embodiments, the present disclosure provides anti-RSV antibody formulations described herein wherein, after storing the formulation at 2-8°C for 12 months, the percentage of intact IgG in the formulation as measured by CE-SDS under non-reducing conditions is >97 %.

In additional embodiments, the present disclosure provides anti-RSV antibody formulations described herein wherein after storing the formulation at 2-8°C for 12 months, the percentage of monomer measured by HP-SEC is >98.5%.

In additional embodiments, the present disclosure provides anti-RSV antibody formulations described herein wherein after storage of the formulation at 2-8°C for 12 months, the percentage of high molecular weight particles measured by HP-SEC is <1.5%.

In further embodiments, the present disclosure provides anti-RSV antibody formulations described herein wherein after storage of the formulation at 25° C. for 12 months, the percentage of monomer measured by HP-SEC is >98.0%.

In further embodiments, the present disclosure provides anti-RSV antibody formulations described herein wherein after storage of the formulation at 25° C. for 6 months, the percentage of high molecular weight particles measured by HP-SEC is <2%.

In further embodiments, the present disclosure provides anti-RSV antibody formulations described herein wherein, after storing the formulation at 40° C. for 3 months, the percentage of monomer as measured by HP-SEC is >94.0%, >94.5 % or >95.0%.

In further embodiments, the present disclosure provides anti-RSV antibody formulations described herein wherein, after storing the formulation at 40° C. for 3 months, the percentage of high molecular weight species measured by HP-SEC is <5.5%, <5. 0% or <4.4%.

V. Specific Aspects and Embodiments of the Present Invention In one aspect, the present disclosure provides an anti-RSV antibody composition comprising: a) an anti-RSV antibody or antigen binding fragment thereof at a concentration of about 50 mg/ml to about 250 mg/ml; b) a buffer at a concentration of about 5 mM to about 20 mM; c) a stabilizer selected from the group consisting of: (i) a non-reducing sugar at a concentration of about 6% to about 8% on a mass-volume (w/v) basis; (ii) L-arginine or a pharmaceutically acceptable L-arginine salt at a concentration of about 25 mM to about 75 mM; L-proline or a pharmaceutically acceptable L-proline salt at a concentration of about 25 mM to about 75 mM; L-glutamate or a pharmaceutically acceptable salt of L-glutamate at a concentration of about 25 mM to about 75 mM; L-lysine or a pharmaceutically acceptable L-lysine salt at a concentration of about 25 mM to about 75 mM, or a mixture of such amino acids, and (iii) a chelator at a concentration of about 25 μM to 75 μM; d) a nonionic surfactant at a concentration of about 0.01% to about 0.10%; and optionally e) an antioxidant at a concentration of about 1 mM to about 20 mM.

In another aspect, the present disclosure provides an anti-RSV antibody composition comprising: a) an anti-RSV antibody or antigen binding fragment thereof at a concentration of about 50 mg/ml to about 150 mg/ml; b) histidine at a concentration of about 5 mM to about 20 mM; c) sucrose at a concentration of from about 6% to about 8% in a mass-volume ratio (w/v); d) an excipient selected from the following: L-arginine or a pharmaceutically acceptable L-arginine salt at a concentration of about 25 mM to about 75 mM, L-lysine or a pharmaceutically acceptable L-lysine salt at a concentration from about 25 mM to about 75 mM , a mixture of L-arginine (HCl) and L-glutamate at a concentration of about 25 mM to about 75 mM and a mixture of L-arginine (HCl) and L-lysine (HCl) at a concentration of about 25 mM to about 75 mM; e) DTPA at a concentration of about 25 to 75 μM; f) polysorbate 80 at a concentration of about 0.01% to about 0.10%; and optionally g) L-methionine at a concentration of about 1 mM to about 20 mM.

In another aspect, the present disclosure provides an anti-RSV antibody composition comprising: a) an anti-RSV antibody or antigen binding fragment thereof at a concentration of about 100 mg/ml; b) histidine at a concentration of about 10 mM; c) sucrose at a concentration of 7% in a mass-volume ratio (w/v); d) an excipient selected from L-arginine (HCl) at a concentration of 50 mM, L-lysine (HCl) at a concentration of 50 mM, a mixture of L-arginine at a concentration of 25 mM and L-glutamate at a concentration of 25 mM and a mixture of L-arginine at a concentration of 25 mM (HCl) and L-lysine (HCl) at a concentration of 25 mM; f) DTPA at a concentration of 50 μM; f) polysorbate 80 at a concentration of 0.02% and optionally g) L-methionine at a concentration of 1 mM to 20 mM.

In another aspect, the present disclosure provides an anti-RSV antibody composition comprising: a) an anti-RSV antibody or antigen binding fragment thereof at a concentration of about 100 mg/ml; b) histidine at a concentration of about 10 mM; c) sucrose at a concentration of 7% in a mass-volume ratio (w/v); d) a mixture of L-arginine at a concentration of 25 mM (HCl) and L-lysine (HCl) at a concentration of 25 mM; e) DTPA at a concentration of 50 µM and f) polysorbate 80 at a concentration of 0.02%.

In another aspect, the concentration of the anti-RSV antibody or antigen-binding fragment thereof is from about 50 mg/ml to about 150 mg/ml. In another aspect, the concentration of the anti-RSV antibody or antigen-binding fragment thereof is from about 75 mg/ml to about 125 mg/ml. In another aspect, the concentration of the anti-RSV antibody or antigen-binding fragment thereof is from 50 mg/ml to 150 mg/ml. In another aspect, the concentration of the anti-RSV antibody or antigen-binding fragment thereof is from 75 mg/ml to 125 mg/ml. In another aspect, the concentration of the anti-RSV antibody or antigen-binding fragment thereof is 100 mg/ml. In another aspect, the concentration of the anti-RSV antibody or antigen-binding fragment thereof is 75 mg/ml. In another aspect, the concentration of the anti-RSV antibody or antigen-binding fragment thereof is 50 mg/ml.

In another aspect, the composition has a pH value of from about 5.5 to about 6.5. According to another aspect, the composition has a pH value from 5.5 to 6.5. According to another aspect, the composition has a pH value of 6.0.

In another aspect, the buffer is histidine. In another aspect, the concentration of histidine is from about 1 mM to about 20 mM. In another aspect, the concentration of histidine is from about 5 mM to about 15 mM. According to another aspect, the concentration of histidine is from 1 mM to 20 mM. According to another aspect, the concentration of histidine is from 5 mM to 15 mM. In another aspect, the histidine concentration is 10 mM.

According to another aspect, the composition comprises a stabilizer selected from a non-reducing sugar; L-arginine or a pharmaceutically acceptable salt of L-arginine; L-proline or a pharmaceutically acceptable salt of L-proline; L-glutamate or a pharmaceutically acceptable salt of L-glutamate; L-lysine or a pharmaceutically acceptable salt of L-lysine or a mixture of such amino acids. In another aspect, the composition contains an excipient selected from L-arginine (HCl) at a concentration of about 50 mM, L-lysine (HCl) at a concentration of about 50 mM, a mixture of L-arginine at a concentration of about 25 mM and L-glutamate at a concentration of about 25 mM and a mixture of L-arginine at a concentration of about 25 mM (HCl) and L-lysine (HCl) at a concentration of about 25 mM.

In another aspect, the composition contains a chelator that is DTPA at a concentration of about 25 μM to about 75 μM. In another aspect, the composition contains a chelator that is DTPA at a concentration of about 50 μM. In another aspect, the composition contains a chelator that is DTPA at a concentration of 50 μM.

In another aspect, the composition contains a nonionic surfactant that is PS80 at a concentration of from about 0.01% to about 0.10% on a weight-volume basis. In another aspect, the composition contains a nonionic surfactant that is PS80 in concentration of about 0.02% w/v. In another aspect, the formulation contains a nonionic surfactant that is PS80 at a concentration of 0.02% w/v

In another aspect, the composition contains an antioxidant that is L-methionine at a concentration of about 1 mM to about 20 mM. In another aspect, the composition contains an antioxidant that is L-methionine at a concentration of about 10 mM. In another aspect, the composition contains an antioxidant that is L-methionine at a concentration of 10 mM.

In another aspect, the present disclosure provides an anti-RSV antibody composition comprising: a) an anti-RSV antibody or antigen binding fragment thereof at a concentration of about 50 mg/ml to about 250 mg/ml; b) histidine at a concentration of about 5 mM to about 20 mM; c) sucrose at a concentration of about 4% to about 8% w/v; d) an excipient selected from the group consisting of the following: L-arginine or a pharmaceutically acceptable L-arginine salt at a concentration of about 25 mM to about 75 mM, L-lysine or a pharmaceutically acceptable L-lysine salt at a concentration of about 25 mM to about 75 mM, a mixture of L-arginine or a pharmaceutically acceptable salt of L-arginine at a concentration of about 25 to 75 mM and L-glutamate or a pharmaceutically acceptable salt of L-glutamate at a concentration of about 25 mM to about 75 mM and a mixture of L-arginine or a pharmaceutically acceptable L-arginine salt at a concentration of about 25 mM to about 75 mM and L-lysine or a pharmaceutically acceptable L-lysine salt at a concentration from about 25 mM to about 75 mM and e) polysorbate 80 at a concentration of about 0.01 % to about 0.10% w/v.

In one embodiment, the anti-RSV antibody composition comprises: a) an anti-RSV antibody or antigen-binding fragment thereof at a concentration of about 50 mg/ml to about 250 mg/ml; b) histidine at a concentration of about 5 mM to about 20 mM; c) sucrose at a concentration of about 4% to about 8% w/v; d) L-arginine or a pharmaceutically acceptable L-arginine salt at a concentration of about 25 mM to about 75 mM; and e) polysorbate 80 at a concentration of about 0.01% to about 0.10% w/v. In one embodiment, the anti-RSV antibody composition comprises: a) an anti-RSV antibody or an antigen-binding fragment thereof at a concentration of 50 mg/ml to 250 mg/ml; b) histidine at a concentration of 5 mM to 20 mM; c) sucrose at a concentration of 4% to 8% w/v; d) L-arginine or a pharmaceutically acceptable L-arginine salt at a concentration of 25 mM to 75 mM; and e) polysorbate 80 at a concentration of 0.01% to 0.10% w/v.

In one embodiment, the anti-RSV antibody composition comprises: a) an anti-RSV antibody or antigen-binding fragment thereof at a concentration of about 50 mg/ml to about 250 mg/ml; b) histidine at a concentration of about 5 mM to about 20 mM; c) sucrose at a concentration of about 4% to about 8% w/v; d) L-lysine or a pharmaceutically acceptable L-lysine salt at a concentration of about 25 mM to about 75 mM; and e) polysorbate 80 at a concentration of about 0.01% to about 0.10% w/v. In one embodiment, the anti-RSV antibody composition comprises: a) an anti-RSV antibody or antigen-binding fragment at a concentration of 50 mg/ml to 250 mg/ml; b) histidine at a concentration of 5 mM to 20 mM; c) sucrose at a concentration of 4% to 8% w/v; d) L-lysine or a pharmaceutically acceptable L-lysine salt at a concentration of 25 mM to 75 mM; and e) polysorbate 80 at a concentration of 0.01% to 0.10% w/v.

In one embodiment, the anti-RSV antibody composition comprises: a) an anti-RSV antibody or antigen-binding fragment thereof at a concentration of about 50 mg/ml to about 250 mg/ml; b) histidine at a concentration of about 5 mM to about 20 mM; c) sucrose at a concentration of about 4% to about 8% w/v; d) a mixture of L-arginine or a pharmaceutically acceptable salt of L-arginine at a concentration of about 25 mM to about 75 mM and L-glutamate or a pharmaceutically acceptable salt of L-glutamate at a concentration of about 25 mM to about 75 mM and e) polysorbate 80 b concentrations from about 0.01% to about 0.10% w/v. In one embodiment, the anti-RSV antibody composition comprises: a) an anti-RSV antibody or an antigen-binding fragment thereof at a concentration of 50 mg/ml to 250 mg/ml; b) histidine at a concentration of 5 mM to 20 mM; c) sucrose at a concentration of 4% to 8% w/v; d) a mixture of L-arginine or a pharmaceutically acceptable salt of L-arginine at a concentration of 25 mM to 75 mM and L-glutamate or a pharmaceutically acceptable salt of L-glutamate at a concentration of 25 mM to 75 mM and e) polysorbate 80 at a concentration of 0. 01% to 0.10% w/v.

In one embodiment, the anti-RSV antibody composition comprises: a) an anti-RSV antibody or antigen-binding fragment thereof at a concentration of about 50 mg/ml to about 250 mg/ml; b) histidine at a concentration of about 5 mM to about 20 mM; c) sucrose at a concentration of about 4% to about 8% w/v; d) a mixture of L-arginine or a pharmaceutically acceptable salt of L-arginine at a concentration of about 25 mM to about 75 mM and L-lysine or a pharmaceutically acceptable salt of L-lysine at a concentration of about 25 mM to about 75 mM and e) polysorbate 80 in concentrations from about 0.01% to about 0.10% w/v. In one embodiment, the anti-RSV antibody composition comprises: a) an anti-RSV antibody or an antigen-binding fragment thereof at a concentration of 50 mg/ml to 250 mg/ml; b) histidine at a concentration of 5 mM to 20 mM; c) sucrose at a concentration of 4% to 8% w/v; d) a mixture of L-arginine or a pharmaceutically acceptable salt of L-arginine at a concentration from 25 mM to 75 mM and L-lysine or a pharmaceutically acceptable salt of L-lysine at a concentration from 25 mM to 75 mM and e) polysorbate 80 at a concentration from 0. 01% to 0.10% w/v.

In yet another aspect, the present disclosure provides an anti-RSV antibody composition comprising: a) an anti-RSV antibody or antigen binding fragment thereof at a concentration of about 50 mg/ml to about 250 mg/ml; b) histidine at a concentration of about 5 mM to about 100 mM; c) sucrose at a concentration of about 4% to about 8% w/v; and d) polysorbate 80 at a concentration of about 0.01% to about 0.10% w/v. In one embodiment, the present disclosure provides an anti-RSV antibody composition comprising: a) an anti-RSV antibody or antigen binding fragment thereof at a concentration of about 50 mg/ml to 250 mg/ml; b) histidine at a concentration of 5 mM to 100 mM; c) sucrose at a concentration of 4% to 8% w/v; and d) polysorbate 80 at a concentration of 0.01% to 0.10% w/v.

In one aspect, the anti-RSV antibody composition comprises: a) an anti-RSV antibody or antigen-binding fragment thereof at a concentration of about 50 mg/ml to about 250 mg/ml; b) histidine at a concentration of about 5 mM to about 100 mM; c) sucrose at a concentration of about 4% to about 8% w/v; d) L-arginine or a pharmaceutically acceptable L-arginine salt at a concentration of about 25 mM to about 75 mM; and e) polysorbate 80 at a concentration of about 0.01% to about 0.10% w/v. In one embodiment, the anti-RSV antibody composition comprises: a) an anti-RSV antibody or an antigen-binding fragment thereof at a concentration of about 50 mg/ml to 250 mg/ml; b) histidine at a concentration of 5 mM to 100 mM; c) sucrose at a concentration of 4% to 8% w/v; d) L-arginine or a pharmaceutically acceptable L-arginine salt at a concentration of 25 mM to 75 mM; and e) polysorbate 80 at a concentration of 0.01% to 0.10% w/v.

In one aspect, the anti-RSV antibody composition comprises: a) an anti-RSV antibody or antigen-binding fragment thereof at a concentration of about 50 mg/ml to about 250 mg/ml; b) histidine at a concentration of about 5 mM to about 100 mM; c) sucrose at a concentration of about 4% to about 8% w/v; d) L-lysine or a pharmaceutically acceptable L-lysine salt at a concentration of about 25 mM to about 75 mM; and e) polysorbate 80 at a concentration of about 0.01% to about 0.10% w/v. In one embodiment, the anti-RSV antibody composition comprises: a) an anti-RSV antibody or an antigen-binding fragment thereof at a concentration of 50 mg/ml to 250 mg/ml; b) histidine at a concentration of 5 mM to 100 mM; c) sucrose at a concentration of 4% to 8% w/v; d) L-lysine or a pharmaceutically acceptable L-lysine salt at a concentration of 25 mM to 75 mM; and e) polysorbate 80 at a concentration of 0.01% to 0.10% w/v.

In one aspect, the anti-RSV antibody composition comprises: a) an anti-RSV antibody or antigen-binding fragment thereof at a concentration of about 50 mg/ml to about 250 mg/ml; b) histidine at a concentration of about 5 mM to about 100 mM; c) sucrose at a concentration of about 4% to about 8% w/v; d) a mixture of L-arginine or a pharmaceutically acceptable salt of L-arginine at a concentration of about 25 mM to about 75 mM and L-glutamate or a pharmaceutically acceptable salt of L-glutamate at a concentration of about 25 mM to about 75 mM and e) polysorbate 80 b concentrations from about 0.01% to about 0.10% w/v. In one embodiment, the anti-RSV antibody composition comprises: a) an anti-RSV antibody or an antigen-binding fragment thereof at a concentration of about 50 mg/ml to 250 mg/ml; b) histidine at a concentration of 5 mM to 100 mM; c) sucrose at a concentration of 4% to about 8% w/v; d) a mixture of L-arginine or a pharmaceutically acceptable salt of L-arginine at a concentration of 25 mM to 75 mM and L-glutamate or a pharmaceutically acceptable salt of L-glutamate at a concentration of 25 mM to 75 mM and e) polysorbate 80 at a concentration of 0. 01% to 0.10% w/v.

In one aspect, the anti-RSV antibody composition comprises: a) an anti-RSV antibody or antigen-binding fragment thereof at a concentration of about 50 mg/ml to about 250 mg/ml; b) histidine at a concentration of about 5 mM to about 100 mM; c) sucrose at a concentration of about 4% to about 8% w/v; d) a mixture of L-arginine or a pharmaceutically acceptable salt of L-arginine at a concentration of about 25 mM to about 75 mM and L-lysine or a pharmaceutically acceptable salt of L-lysine at a concentration of about 25 mM to about 75 mM and e) polysorbate 80 b concentrations from about 0.01% to about 0.10% w/v. In one embodiment, the anti-RSV antibody composition comprises: a) an anti-RSV antibody or antigen-binding fragment thereof at a concentration of about 50 mg/ml to 250 mg/ml; b) histidine at a concentration of 5 mM to 100 mM; c) sucrose at a concentration of 4% to 8% w/v; d) a mixture of L-arginine or a pharmaceutically acceptable salt of L-arginine at a concentration of 25 mM to 75 mM and L-lysine or a pharmaceutically acceptable salt of L-lysine at a concentration of 25 mM to 75 mM and e) polysorbate 80 at a concentration of 0. 01% to 0.10% w/v.

In some embodiments of any of the above aspects and embodiments, the concentration of histidine is from about 70 mM to about 90 mM. In some embodiments, the histidine concentration is between 70 mM and 90 mM. In some embodiments, the histidine concentration is about 80 mM. In some embodiments, the histidine concentration is 80 mM.

In some embodiments of the above aspects and embodiments, the anti-RSV antibody composition comprises an anti-RSV antibody or an antigen-binding fragment thereof at a concentration of about 125 mg/mL to about 175 mg/mL. In one embodiment, the anti-RSV antibody composition comprises an anti-RSV antibody or an antigen-binding fragment thereof at a concentration of 125 mg/ml to 175 mg/ml. In one embodiment, the anti-RSV antibody composition contains an anti-RSV antibody or an antigen-binding fragment thereof at a concentration of about 150 mg/ml. In one embodiment, the anti-RSV antibody composition comprises an anti-RSV antibody or an antigen-binding fragment thereof at a concentration of 150 mg/ml.

In some embodiments of the above aspects and embodiments, the anti-RSV antibody composition comprises an anti-RSV antibody or an antigen-binding fragment thereof at a concentration of about 150 mg/ml to about 200 mg/ml. In some embodiments, the anti-RSV antibody composition comprises an anti-RSV antibody or an antigen-binding fragment thereof at a concentration of 150 mg/mL to 200 mg/mL. In some embodiments, the anti-RSV antibody composition comprises an anti-RSV antibody or an antigen-binding fragment thereof at a concentration of about 175 mg/ml. In some embodiments, the anti-RSV antibody composition comprises an anti-RSV antibody or an antigen-binding fragment thereof at a concentration of 175 mg/mL.

In some embodiments of the above aspects and embodiments, the anti-RSV antibody composition comprises an anti-RSV antibody or an antigen-binding fragment thereof at a concentration of about 175 mg/ml to about 225 mg/ml. In some embodiments, the anti-RSV antibody composition comprises an anti-RSV antibody or an antigen-binding fragment thereof at a concentration of 175 mg/mL to 225 mg/mL. In some embodiments, the anti-RSV antibody composition comprises an anti-RSV antibody or an antigen-binding fragment thereof at a concentration of about 200 mg/ml. In some embodiments, the anti-RSV antibody composition comprises an anti-RSV antibody or an antigen-binding fragment thereof at a concentration of 200 mg/ml.

In some embodiments of the above aspects and embodiments, the anti-RSV antibody composition further comprises diethylenetriamine pentaacetate (DTPA) at a concentration of about 25 to about 75 μM. In some embodiments, the anti-RSV antibody formulation further comprises diethylenetriamine pentaacetate (DTPA) at a concentration of 25 to 75 μM.

In some embodiments of the above aspects and embodiments, the anti-RSV antibody composition further comprises L-methionine at a concentration of about 1 mM to about 20 mM. In some embodiments, the anti-RSV antibody composition further comprises L-methionine at a concentration of 1 mM to 20 mM.

In some embodiments, the pH of the anti-RSV antibody formulation is between 5.5 and 6.5. In some embodiments, the pH of the anti-RSV antibody formulation is about 6.0. In some embodiments, the pH of the anti-RSV antibody formulation is 6.0.

In some embodiments, the anti-RSV antibody comprises CDRs having the amino acid sequences of SEQ ID NO: 1-6. In some embodiments, the anti-RSV antibody comprises a heavy chain variable region having the amino acid sequence of SEQ ID NO: 7 and a light chain variable region having the amino acid sequence of SEQ ID NO: 8. In some embodiments, the anti-RSV antibody comprises an immunoglobulin heavy chain , consisting of the amino acid sequence shown in SEQ ID NO: 9, and an immunoglobulin light chain, consisting of the amino acid sequence shown in SEQ ID NO: 10.

In one aspect, the present disclosure provides a method of treating or preventing an RSV infection in a human patient in need thereof, comprising: administering to the patient an effective amount of an anti-RSV antibody composition of any of the above aspects and embodiments.

In some embodiments of the foregoing method, an effective amount of the anti-RSV antibody formulation comprises a fixed dose of the anti-RSV antibody or antigen-binding fragment thereof, ranging from about 10 mg to about 150 mg. In some embodiments, an effective amount of the anti-RSV antibody formulation comprises a fixed dose of the anti-RSV antibody or antigen-binding fragment thereof, ranging from 10 mg to 150 mg. In some embodiments, an effective amount of the anti-RSV antibody formulation provides a fixed dose of the anti-RSV antibody or antigen binding fragment thereof, ranging from about 25 mg to about 125 mg. In some embodiments, an effective amount of the anti-RSV antibody formulation comprises a fixed dose of the anti-RSV antibody or antigen-binding fragment thereof, ranging from 25 mg to 125 mg. In some embodiments, an effective amount of the anti-RSV antibody formulation provides a fixed dose of the anti-RSV antibody or antigen binding fragment thereof, ranging from about 50 mg to about 100 mg. In some embodiments, an effective amount of the anti-RSV antibody formulation comprises a fixed dose of the anti-RSV antibody or antigen binding fragment thereof, ranging from 50 mg to 100 mg. In some embodiments, an effective amount of the anti-RSV antibody formulation provides a fixed dose of the anti-RSV antibody or antigen-binding fragment thereof of about 100 mg. In some embodiments, an effective amount of the anti-RSV antibody formulation provides a fixed dose of the anti-RSV antibody or antigen-binding fragment thereof of about 100 mg.

In some embodiments of the above method, the anti-RSV antibody composition is administered intramuscularly.

In some embodiments, the present disclosure provides an anti-RSV antibody formulation in accordance with any of the above aspects or embodiments for use in a method of treating or preventing RSV infection in a human patient.

In some embodiments, the present disclosure provides for use of an anti-RSV antibody composition in accordance with any of the above aspects or embodiments in the manufacture of a medicament for treating or preventing RSV infection in a human patient.

According to some embodiments of the present invention, any of the compositions described herein is in an aqueous solution. In alternative embodiments, the present invention provides lyophilized formulations prepared by lyophilizing an aqueous formulation to produce a reconstituted formulation of the present invention, as discussed in more detail below.

VI. Lyophilized pharmaceutical compositions

Lyophilized formulations of therapeutic proteins (eg, anti-RSV antibody or antigen-binding fragment thereof) provide several advantages. Lyophilized formulations generally have better chemical stability than solution formulations and thus increase half-life. The lyophilized formulation may also be reconstituted at varying concentrations depending on clinical factors such as route of administration or dosage. For example, the lyophilized formulation may be reconstituted at a high concentration (ie, in a small volume) if required for subcutaneous administration, or at a lower concentration if administered intravenously. High concentrations may also be necessary if a high dosage is required for a particular subject, especially when administered subcutaneously where the volume of injection must be minimized. One such lyophilized antibody formulation is disclosed in US Pat. No. 6,267,958, which is incorporated herein by reference in its entirety. Lyophilized formulations of another therapeutic protein are disclosed in US Pat. No. 7,247,707, which is incorporated herein by reference in its entirety.

Typically, the lyophilized formulation is prepared in anticipation of reconstitution at a high concentration of drug product (DP, in an exemplary embodiment, an anti-RSV antibody or antigen binding fragment thereof), that is, in anticipation of reconstitution in a small volume of water. Subsequent dilution with water or isotonic buffer can easily be used to dilute the DP to a lower concentration. Typically, the excipients are included in the lyophilized formulation of the present invention in amounts that will result in an approximately isotonic composition when reconstituted at a high DP concentration, for example, for subcutaneous administration. Reconstitution in a larger volume of water to obtain a lower concentration of DP will necessarily reduce the tonicity of the reconstituted solution, but such reduction may have little effect for non-subcutaneous administration, such as intravenous administration. If isotonicity is desired at a lower DP concentration, the lyophilized powder can be reconstituted in a standard small volume of water and then diluted with an isotonic diluent such as 0.9% sodium chloride.

In one embodiment of the present invention, an anti-RSV antibody (or an antigen-binding fragment thereof) is formulated as a lyophilized powder for reconstitution and use for intravenous, subcutaneous or intramuscular administration. In some embodiments, the antibody (or antigen binding fragment thereof) is provided in an amount of about 50 mg/vial and is reconstituted with sterile water for injection prior to use. If desired, the reconstituted antibody can be aseptically reconstituted with 0.9% Sodium Chloride Injection USP in a sterile IV container. In some embodiments, the target pH of the reconstituted formulation is 5.5 ± 0.5. In various embodiments, the lyophilized formulation of the present invention can reconstitute the anti-RSV antibody to high concentrations, such as about 20, 25, 30, 40, 50, 60, 75, 100, 125, 150, 175 or more mg/ml.

Lyophilized formulations, by definition, are essentially dry and therefore the concept of concentration is not useful in describing them. Describing a lyophilized formulation in terms of the mass of components in a unit dose vial is more applicable, but is problematic because it varies for different doses or vial sizes. When describing the lyophilized formulations of the present invention, it is useful to express the amount of a component as the ratio of the weight of the component to the weight of the drug substance (DS) in the same sample (eg, vial). This ratio can be expressed as a percentage. Such ratios reflect the intrinsic properties of the lyophilized formulations of the present invention, regardless of vial size, dosage, and reconstitution protocol.

In other embodiments, the lyophilized composition of an anti-RSV antibody or antigen binding fragment thereof is defined in terms of the pre-lyophilization solution used to prepare the lyophilized composition, such as the pre-lyophilization solution. In one embodiment, the lyophilization solution contains the antibody or antigen binding fragment thereof at a concentration of about 10 mg/ml, about 25 mg/ml, or about 50 mg/ml. Such pre-lyophilization solutions may have a pH value of 4.4-5.2 (including about 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5 1 and 5.2), for example, preferably about pH 4.8 or about 5.5.

In other embodiments, the lyophilized formulation of an anti-RSV antibody or antigen-binding fragment thereof is defined as a reconstituted solution prepared from the lyophilized formulation.

Reconstituted solutions may contain the antibody or antigen binding fragment thereof at concentrations of about 50, 60, 75, 80, 90 or 100 mg/ml or higher concentrations such as 150 mg/ml, 167 mg/ml, 200 mg/ml or about 250 mg/ml. Such reconstituted solutions may have a pH value of about 5.5 or a pH range from about 5.0 to about 6.0.

The lyophilized compositions of the present invention are prepared by lyophilizing (freeze-drying) a pre-lyophilization solution. Freeze drying is carried out by freezing the composition and subsequent sublimation of water at a temperature suitable for primary drying. Under these conditions, the temperature of the product is below the eutectic point or temperature of destruction of the composition. Typically, the primary drying shelf temperature will be in the range of about -30 to 25°C (assuming the product remains frozen during primary drying) with a suitable pressure typically in the range of about 50 to 250 mTorr. The composition, size and type of container containing the sample (eg, glass vial), and volume of liquid will determine the time required for drying, which can vary from several hours to several days (eg, 40-60 hours). The secondary drying step can be carried out at a temperature of about 0-40°C, depending primarily on the type and size of the container and the type of protein used. The secondary drying time is determined by the desired residual moisture level of the product and typically takes at least about 5 hours. Typically, the moisture content of the lyophilized composition is less than about 5%, and preferably less than about 3%. The pressure can be the same as during the primary drying stage. Freeze-drying conditions can be varied depending on the composition and size of the bottle.

In some cases, it may be desirable to lyophilize the protein formulation in the container in which the protein reconstitution is to occur to avoid a transfer step. The container in this case can be, for example, a bottle with a volume of 3, 5, 10, 20, 50 or 100 ml.

The lyophilized formulations of the present invention are reconstituted before administration. The protein can be reconstituted at concentrations of about 10, 15, 20, 25, 30, 40, 50, 60, 75, 80, 90 or 100 mg/ml or higher concentrations such as 150 mg/ml, 200 mg/ml, 250 mg/ml or 300 mg/ml to about 500 mg/ml. High protein concentrations are particularly useful when subcutaneous delivery of the reconstituted formulation is contemplated. However, for other routes of administration, such as intravenous administration, lower protein concentrations (eg, about 5 to 50 mg/ml) may be desirable.

Reduction typically occurs at around 25°C to ensure complete hydration, although other temperatures can be used if desired. The time required for reconstitution will depend, for example, on the type of diluent, the amount of excipient(s) and protein. Exemplary diluents include sterile water, bacteriostatic water for injection (BWFI), pH buffer solution (eg, phosphate buffered saline), sterile saline, Ringer's solution, or dextrose solution.

The present disclosure provides a liquid formulation of an anti-RSV antibody reconstituted from a lyophilized formulation, the reconstituted solution containing: a) an anti-RSV antibody or antigen binding fragment thereof at a concentration of about 50 mg/ml to about 150 mg/ml; b) histidine at a concentration of about 5 mM to about 20 mM; c) sucrose at a concentration of from about 6% to about 8% in a mass-volume ratio (w/v); d) an excipient selected from the following: L-arginine or a pharmaceutically acceptable salt of L-arginine in a concentration of about 25 mM to about 75 mM, L-proline or a pharmaceutically acceptable salt of L-proline from about 25 mM to about 75 mM, L β-glutamate or a pharmaceutically acceptable salt of L-glutamate at a concentration of about 25 mM to about 75 mM, L-lysine or a pharmaceutically acceptable salt of L-lysine at a concentration of about 25 mM to about 75 mM, a mixture of L-arginine and L-glutamate in concentrations of about 25 mM to about 75 mM and a mixture of L-arginine (HCl) and L-lysine (HCl) at a concentration of about 25 mM to about 75 mM; e) DTPA at a concentration of about 25 μM to 75 μM; f) polysorbate 80 at a concentration of about 0.01% to about 0.10%; and optionally g) L-methionine at a concentration of about 1 mM to about 20 mM.

The present disclosure provides a liquid formulation of an anti-RSV antibody reconstituted from a lyophilized formulation, the reconstituted solution containing: a) an anti-RSV antibody or antigen binding fragment thereof at a concentration of about 100 mg/ml; b) histidine at a concentration of about 10 mM; c) sucrose at a concentration of 7% in a mass-volume ratio (w/v); d) an excipient selected from L-arginine (HCl) at a concentration of 50 mM, L-lysine (HCl) at a concentration of 50 mM, a mixture of L-arginine at a concentration of 25 mM and L-glutamate at a concentration of 25 mM and a mixture of L-arginine at a concentration of 25 mM (HCl) and L-lysine (HCl) at a concentration of 25 mM; f) DTPA at a concentration of 50 μM; f) polysorbate 80 at a concentration of 0.02% and optionally g) L-methionine at a concentration of 1 mM to 20 mM.

The present disclosure provides a liquid formulation of an anti-RSV antibody reconstituted from a lyophilized formulation, the reconstituted solution containing: a) an anti-RSV antibody or antigen binding fragment thereof at a concentration of about 100 mg/ml; b) histidine at a concentration of about 10 mM; c) sucrose at a concentration of 7% in a mass-volume ratio (w/v); d) a mixture of L-arginine at a concentration of 25 mM (HCl) and L-lysine (HCl) at a concentration of 25 mM; e) DTPA at a concentration of 50 µM and f) polysorbate 80 at a concentration of 0.02%.

The present disclosure also provides a liquid anti-RSV antibody formulation reconstituted from a lyophilized formulation, the reconstituted solution containing: a) an anti-RSV antibody or antigen binding fragment thereof at a concentration of about 50 mg/ml to about 250 mg/ml; b) histidine at a concentration of about 5 mM to about 20 mM; c) sucrose at a concentration of about 4% to about 8% w/v; d) an excipient selected from the group consisting of the following: L-arginine or a pharmaceutically acceptable L-arginine salt at a concentration of about 25 mM to about 75 mM, L-lysine or a pharmaceutically acceptable L-lysine salt at a concentration of about 25 mM to about 75 mM, a mixture of L-arginine or a pharmaceutically acceptable salt of L-arginine at a concentration of about 25 to 75 mM and L-glutamate or a pharmaceutically acceptable salt of L-glutamate at a concentration of about 25 mM to about 75 mM and a mixture of L-arginine or a pharmaceutically acceptable L-arginine salt at a concentration of about 25 mM to about 75 mM and L-lysine or a pharmaceutically acceptable L-lysine salt at a concentration from about 25 mM to about 75 mM and e) polysorbate 80 at a concentration of about 0.01 % to about 0.10% w/v.

VII. Liquid pharmaceutical compositions

A liquid antibody formulation can be prepared by exchanging the buffer of a drug (eg, anti-RSV antibody) that is in liquid form with the desired buffer as a final step in the purification process. According to this embodiment, there is no lyophilization step. The drug substance in the final buffer is concentrated to the desired concentration. Excipients such as sucrose, methionine and polysorbate 80 are added to the drug substance and diluted with an appropriate buffer to the final protein concentration. The final drug formulation is filtered, for example using 0.22 µm filters, and dispensed into the final container (eg glass vials or syringes). An example of such a liquid formulation is a final liquid formulation containing a) an anti-RSV antibody or an antigen-binding fragment thereof at a concentration of about 100 mg/ml; b) histidine at a concentration of about 10 mM; c) sucrose at a concentration of 7% in a mass-volume ratio (w/v); d) a mixture of L-arginine at a concentration of 25 mM (HCl) and L-lysine (HCl) at a concentration of 25 mM; e) DTPA at a concentration of 50 µM and f) polysorbate 80 at a concentration of 0.02%.

VIII. Methods of application

The present invention also provides a method for preventing or treating RSV infection in a subject, the method comprising administering an effective amount of any of the compositions of the present invention; those. any composition described herein to the subject. In some embodiments of this method, the composition is administered intravenously to a subject. In other embodiments, the composition is administered to the subject subcutaneously. In other embodiments, the composition is administered intramuscularly.

In a specific embodiment, a prophylactic, therapeutic, or pharmaceutical composition of the present invention is administered to a mammal, preferably a human, to treat, prevent, or alleviate one or more symptoms associated with RSV infection, in an amount effective to reduce RSV titers. In accordance with this embodiment, an effective amount of an anti-RSV antibody or antibody fragment reduces RSV titers in the lungs as measured, for example, by the concentration of RSV in sputum samples or lung lavage fluid of a mammal. In another embodiment, a prophylactic, therapeutic or pharmaceutical composition comprising an anti-RSV antibody of the present invention or fragments thereof is administered to a mammal, preferably a human, to treat, prevent or alleviate symptoms associated with RSV infection in an amount effective to neutralize RSV and/or or blocking RSV infection in mammals.

The compositions of the present invention can also be used immunotherapeutically for RSV disease in both humans and other animals. The term "immunotherapeutic" or "immunotherapy" as used herein in combination with an anti-RSV antibody or antigen-binding fragments thereof of the present invention means both prophylactic and therapeutic administration as well as passive immunization with substantially purified polypeptide products, as well as gene therapy by transferring polynucleotide sequences encoding the product or part thereof. Passive immunization involves the transfer of active humoral immunity or the provision of antibodies to a subject in need. Accordingly, in certain embodiments of the present invention, the present invention provides methods for conveying active humoral immunity and methods for providing anti-RSV antibodies or antigen-binding fragments thereof, such as IgG antibodies, to a patient at risk of RSV infection. Thus, monoclonal antibodies or antigen-binding fragments thereof can be administered to high-risk subjects to reduce the likelihood and/or severity of RSV disease, or administered to subjects already demonstrating active RSV infection.

The present invention also provides a method for modulating or treating at least one disease associated with RSV in an adult or child, cell, tissue, organ, animal or patient, including, but not limited to, lower respiratory tract infections, pneumonia, tracheobronchitis, bronchiolitis, bronchitis and any associated infections or inflammatory disorders, such as, without limitation, at least one or at least one of the following: inflammation associated with systemic inflammatory response syndrome, sepsis syndrome, gram-positive sepsis, gram-negative sepsis, culture-negative sepsis, fungal sepsis, neutropenic fever, urosepsis, meningococcemia, respiratory distress syndrome in adults, allergic rhinitis, perennial rhinitis, bronchial asthma, systemic anaphylaxis, receptor hypersensitivity reaction, chronic obstructive pulmonary disease (COPD), hypersensitivity pneumonitis, granulomas caused by intracellular organisms, drug sensitivity, cachexia, cystic fibrosis, chronic lung diseases of newborns; at least one infectious disease in a cell, tissue, organ, animal or patient, including, without limitation, at least one of the following: acute or chronic bacterial infection, acute and chronic parasitic or infectious processes, including bacterial , viral and fungal infections, HIV infection, HIV neuropathy, meningitis, hepatitis (A, B or C, etc.), septic arthritis, peritonitis, pneumonia, epiglottitis, E. coli 0157:h7, hemolytic-uremic syndrome , thrombolytic thrombocytopenic purpura, malaria, dengue hemorrhagic fever, leishmaniasis, leprosy, toxic shock syndrome, streptococcal myositis, gas gangrene, Mycobacterium tuberculosis, Mycobacterium avium intracellulare, Pneumocystis pneumonia, pelvic inflammatory disease, orchitis, epididymitis, Legionella , Lyme disease, influenza A, Epstein-Barr virus associated hemophagocytic syndrome, vital encephalitis, aseptic meningitis, etc. Such a method may optionally involve administering an effective amount of a composition or pharmaceutical composition comprising at least one anti-RSV antibody or antigenic fragment thereof to a cell, tissue, organ, animal or patient in need of such modulation, treatment or therapy.

In one embodiment, a prophylactic, therapeutic, or pharmaceutical composition comprising an anti-RSV antibody of the present invention or fragments thereof is administered to a mammal, preferably a human, to treat, prevent, or alleviate one or more symptoms associated with RSV infection. In another embodiment, prophylactic, therapeutic, or pharmaceutical compositions containing an anti-RSV antibody of the present invention or fragments thereof are administered to a person with cystic fibrosis, bronchopulmonary dysplasia, congenital heart disease, congenital immunodeficiency or acquired immunodeficiency, or a person who has undergone a transplant (e.g., bone marrow, lung, or hematopoietic stem cell (HSCT)) to treat, prevent, or alleviate one or more symptoms associated with RSV infection.

In another embodiment, prophylactic, therapeutic, or pharmaceutical compositions containing an anti-RSV antibody of the present invention or fragments thereof are administered to a human infant, preferably a premature human infant or a human infant at risk of hospitalization due to RSV infection for the treatment, prevention, or relief of one or more symptoms associated with RSV infection. In yet another embodiment, the prophylactic, therapeutic or pharmaceutical compositions of the present invention are administered to elderly people or people in group homes (eg, nursing homes or rehabilitation centers) or immunocompromised individuals.

According to another embodiment, the present invention provides a method for preventing respiratory infection caused by RSV. In another embodiment, the present invention provides a method for preventing lower respiratory tract infection caused by RSV. In another embodiment, the present invention provides a method for preventing a lower respiratory tract infection caused by RSV A and B strains. In another embodiment, the present invention provides a method for preventing a medically supervised lower respiratory tract infection caused by RSV A and B strains in infants , born at >29 weeks' gestation and <8 months of age at the time of dosing. In another embodiment, the present invention provides a method for preventing RSV lower respiratory tract infection in all infants entering their first RSV season and in children with chronic lung disease or congenital heart disease entering their first and second RSV season.

IX. General methods

Standard techniques in molecular biology are described in Sambrook, Fritsch and Maniatis (1982 & 1989 ^2nd Edition, 2001 ^3rd Edition) Molecular Cloning, A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY; Sambrook and Russell (2001) Molecular Cloning, ^3rd ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY; Wu (1993) Recombinant DNA, Vol. 217, Academic Press, San Diego, CA). Standard methods are also presented in Ausubel, et al. (2001) Current Protocols in Molecular Biology, Vols. 1-4, John Wiley and Sons, Inc. New York, NY, which covers bacterial cloning and DNA mutagenesis (Volume 1), cloning in mammalian cells and yeast (Volume 2), glycoconjugates and protein expression (Volume 3), and bioinformatics (Volume 4).

Methods for protein purification have been described, including immunoprecipitation, chromatography, electrophoresis, centrifugation and crystallization (Coligan, et al. (2000) Current Protocols in Protein Science, Vol. 1, John Wiley and Sons, Inc., New York). Chemical analysis, chemical modification, post-translational modification, production of fusion proteins, protein glycosylation are described (see, for example, Coligan, et al. (2000) Current Protocols in Protein Science, Vol. 2, John Wiley and Sons, Inc., New York; Ausubel, et al (2001) Current Protocols in Molecular Biology, Volume 3, John Wiley and Sons, Inc., NY, NY, pp. 16.0.5-16.22.17; Sigma-Aldrich, Co. (2001) Products for Life Science Research, St. Louis, MO; pp. 45-89; Amersham Pharmacia Biotech (2001) BioDirectory, Piscataway, N.J., pp. 384-391). The production, purification and fragmentation of polyclonal and monoclonal antibodies are described (Coligan, et al. (2001) Current Protocols in Immunology, Vol. 1, John Wiley and Sons, Inc., New York; Harlow and Lane (1999) Using Antibodies, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY; Harlow and Lane, supra). Standard methods are available for characterizing ligand/receptor interactions (see, for example, Coligan, et al. (2001) Current Protocols in Immunology, Vol. 4, John Wiley, Inc., New York).

Monoclonal, polyclonal and humanized antibodies can be prepared (see, for example, Sheperd and Dean (eds.) (2000) Monoclonal Antibodies, Oxford Univ. Press, New York, NY; Kontermann and Dubel (eds.) (2001) Antibody Engineering, Springer-Verlag, New York; Harlow and Lane (1988) Antibodies A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, pp. 139-243; Carpenter, et al. (2000) J. Immunol. 165: 6205; He, et al. (1998) J. Immunol. 160:1029; Tang et al. (1999) J. Biol. Chem. 274: 27371-27378; Baca et al. (1997) J. Biol. Chem. 272:10678-10684; Chothia et al (1989) Nature 342:877-883; Foote and Winter (1992) J Mol Biol 224:487-499; US Patent No. 6329511).

An alternative to humanization is the use of human antibody libraries displayed on phage libraries or human antibody libraries in transgenic mice (Vaughan et al. (1996) Nature Biotechnol. 14:309-314; Barbas (1995) Nature Medicine 1:837-839; Mendez et al. al (1997) Nature Genetics 15:146–156; Hoogenboom and Chames (2000) Immunol. Today 21:371–377; Barbas et al (2001) Phage Display: A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York; Kay et al. (1996) Phage Display of Peptides and Proteins: A Laboratory Manual, Academic Press, San Diego, CA; de Bruin et al. (1999) Nature Biotechnol. 17:397-399).

Antigen purification is not required to form antibodies. Animals can be immunized with cells carrying the antigen of interest. Splenocytes can then be isolated from immunized animals, and the splenocytes can be fused with a myeloma cell line to produce a hybridoma (see, for example, Meyaard et al. (1997) Immunity 7:283-290; Wright et al. (2000) Immunity 13: 233-242; Preston et al, supra; Kaithamana et al (1999) J Immunol 163:5157-5164).

Antibodies can be conjugated, for example, to small drug molecules, enzymes, liposomes, polyethylene glycol (PEG). Antibodies are useful for therapeutic, diagnostic kits or other purposes and include antibodies associated, for example, with dyes, radioisotopes, enzymes or metals, for example colloidal gold (see, for example, Le Doussal et al. (1991) J. Immunol 146:169-175 Gibellini et al (1998) J Immunol 160:3891-3898 Hsing and Bishop (1999) J Immunol 162:2804-2811 Everts et al (2002) J Immunol 168:883-889).

Flow cytometry techniques are available, including fluorescence activated cell sorting (FACS) (see, for example, Owens, et al. (1994) Flow Cytometry Principles for Clinical Laboratory Practice, John Wiley and Sons, Hoboken, NJ; Givan (2001) Flow Cytometry, ^2nd ed.; Wiley-Liss, Hoboken, NJ; Shapiro (2003) Practical Flow Cytometry, John Wiley and Sons, Hoboken, NJ). Fluorescent reagents suitable for modifying nucleic acids are available, including nucleic acid primers and probes, polypeptides and antibodies, for use, for example, as diagnostic reagents (Molecular Probes (2003) Catalog, Molecular Probes, Inc., Eugene, OR; Sigma-Aldrich (2003) Catalog, St. Louis, MO).

Standard techniques for the histology of the immune system have been described (see, for example, Muller-Harmelink (ed.) (1986) Human Thymus: Histopathology and Pathology, Springer Verlag, New York, NY; Hiatt, et al. (2000) Color Atlas of Histology, Lippincott , Williams, and Wilkins, Phila, PA; Louis, et al. (2002) Basic Histology: Text and Atlas, McGraw-Hill, New York, NY).

Software packages and databases are available to determine, for example, antigenic fragments, leader sequences, protein folding, functional domains, glycosylation sites, and sequence alignments (see, for example, GenBank, Vector NTI Suite (Informax, Inc, Bethesda, MD); GCG Wisconsin Package (Accelrys, Inc., San Diego, CA); DECYPHER (TimeLogic Corp., Crystal Bay, Nevada); Menne, et al. (2000) Bioinformatics 16:741-742; Menne, et al. (2000) Bioinformatics Applications Note 16:741-742 Wren, et al (2002) Comput Methods Programs Biomed 68:177-181 von Heijne (1983) Eur J Biochem 133:17-21 von Heijne (1986) Nucleic Acids Res 14:4683–4690).

X. Analytical methods.

Analytical methods suitable for assessing product stability include size exclusion chromatography (SEC), dynamic light scattering (DLS), differential scanning calorimetry (DSC), iso-aspartate quantitation, activity, UV at 340 nm, UV spectroscopy and FTIR . SEC (J. Pharm. Scien., 83:1645-1650, (1994); Pharm. Res., 11:485 (1994); J. Pharm. Bio. Anal., 15:1928 (1997); J. Pharm Bio. Anal., 14:1133-1140 (1986)) measures the percentage of monomer in the product and gives information about the amount of soluble aggregates. DSC (Pharm. Res., 15:200 (1998); Pharm. Res., 9:109 (1982)) provides information on the protein denaturation temperature and glass transition temperature. DLS (American Lab., November (1991)) measures the average diffusion coefficient and provides information on the number of soluble and insoluble aggregates. UV at 340 nm measures the intensity of scattered light at 340 nm and provides information about the amount of soluble and insoluble aggregates. UV spectroscopy measures absorbance at 278 nm and provides information about protein concentration. FTIR (Eur. J. Pharm. Biopharm., 45:231 (1998); Pharm. Res., 12:1250 (1995); J. Pharm. Scien., 85:1290 (1996); J. Pharm. Scien. , 87:1069 (1998)) measures the IR spectrum in one amide region and provides information about the secondary structure of the protein.

The iso-aspartate content of the samples is measured using the Isoaspartate Isoquant Determination System (Promega). The kit uses the enzyme isoaspartyl methyltransferase (PIMT) to specifically detect the presence of isoaspartic acid residues in a target protein. PIMT catalyzes the transfer of a methyl group from S-adenosyl-L-methionine to isoaspartic acid at the α-carboxyl position, thereby forming S-adenosyl-L-homocysteine (SAH). It is a relatively small molecule and can generally be isolated and quantified by reverse phase HPLC using the SAH HPLC standards provided in the kit.

The effectiveness or bioidentity of an antibody can be measured by its ability to bind to its antigen. The specific binding of an antibody to its antigen can be quantified by any method known to those skilled in the art, for example, an immunoassay such as an ELISA (enzyme-linked immunosorbent assay).

All publications cited herein are incorporated by reference for the purpose of describing and disclosing methodologies and materials that may be used in connection with the present invention.

Having described various embodiments of the present invention herein with reference to the accompanying drawings, it is understood that the present invention is not limited to these precise embodiments, and that various changes and modifications may be made by one skilled in the art without departing from the scope or spirit of the present invention. invention as defined in the accompanying claims.

EXAMPLE 1

Stability of anti-RSV antibody formulation at different pH values The present study was performed with a concentration of 100 mg/ml anti-RSV antibody in 10 mM histidine buffer. Sucrose was added to the formulation at a concentration of seven% w/v to increase the volumetric stability (as a stabilizer and nonionic tonicity modifier) of the molecule. Briefly, drug substance (DS) was prepared at a concentration of 100 mg/mL in 10 mM histidine, 7% sucrose at pH 5.5, pH 6.0, and pH 6.5. The formulated DS was bottled and tested for stability at 5°C (ambient humidity), 25°C (60% relative humidity) and 40°C (75% relative humidity) for three months, protected from light.

Samples were assessed by size exclusion chromatography (SEC) for purity, which determined the percentage of monomer, as well as the percentage of high molecular weight particles (HMW) and late elution peaks (LMW particles). Ultra-high performance exclusion chromatography (UP-SEC) was performed by diluting samples (antibodies to RSV) to 1.0 mg/mL in the mobile phase (100 mM sodium phosphate and 100 mM sodium chloride, pH 7.0); flow rate 0.5 ml/min. Diluted samples were injected (5 μL) into a UPLC equipped with a Waters BEN200 column and UV detector. Proteins in the sample were separated by size and detected by UV absorbance at 214 nm.

This study was conducted to examine the effect of pH on the stability of anti-RSV antibody formulation. Based on the results of a heat stability study up to 3 months, there was no noticeable difference between the stability of the 100 mg/mL formulations at pH 5.5, 6.0, and 6.5 for any of the conditions. No changes were seen in UP-SEC % monomer levels at 5°C; however, a small decrease (~2%) was noted at 25°C and a larger decrease (~20%) at 40°C. Based on the results, a pH value of 6.0 was selected for further development (see Table 1).

EXAMPLE 2

Screening (to reduce aggregation) of excipients (amino acids and their mixtures) of the composition of antibodies to RSV

This study compared the stability of anti-RSV antibody in 10 mM histidine (pH=6), 0.02% PS80, 7% sucrose in the presence of (i) 50 mM L-Arg.HCl or (ii) L-Lys .HCl at a concentration of 50 mM or (iii) a mixture of L-Arg at a concentration of 25 mM and L-Glu at a concentration of 25 mM or (iv) a mixture of L-Arg. HCl at a concentration of 25 mM and L-Lys.HCl at a concentration of 25 mM.

These four excipients were tested for reduced aggregation, improved chemical stability (see Example 3) and reduced viscosity (see Example 4). The four excipients studied were:

(i) L-Arg.HCl at a concentration of 50 mM;

(ii) L-Lys.HCl at a concentration of 50 mM;

(iii) a mixture of L-Arg at a concentration of 25 mM and L-Glu at a concentration of 25 mM; and

(iv) a mixture of L-Arg.HCl at a concentration of 25 mM and L-Lys.HCl at a concentration of 25 mM.

The first two excipients were studied to understand the effects of positively charged lysine and arginine in the presence of the chloride counteranion. A third excipient was studied to understand the effect of arginine in the presence of another counteranion (glutamate). The fourth excipient was a mixture of the first two excipients to determine whether the two excipients could play a complementary role. It was hypothesized that this mixture may play a better role than individual excipients, since the mixture of excipients has different pKa and differentially protects the distribution of monoclonal antibody surface changes, thereby increasing repulsive interactions, which in turn reduces aggregation and viscosity.

To evaluate the stability of the formulations, five formulations were bottled and stabilized at 5°C (ambient humidity), 25°C (60% relative humidity) and 40°C (75% relative humidity) for eight weeks, protected from light. place. The five formulations are listed in Table 2 below.

Samples were assessed by size exclusion chromatography (SEC) for purity, and the percentage of monomer, as well as the percentage of high molecular weight particles (HMW) and late elution peaks (LMW particles), were determined. Ultra-high performance exclusion chromatography (UP-SEC) was performed by diluting samples to 1.0 mg/mL (anti-RSV) in mobile phase (100 mM sodium phosphate and 100 mM sodium chloride, pH 7.0); flow rate 0.5 ml/min. Diluted samples were injected (5 μL) into a UPLC equipped with a Waters BEN200 column and UV detector. Proteins in the sample were separated by size and detected by UV absorbance at 214 nm.

UPSEC data for estimating high molecular weight (HMW or aggregate) levels, % monomers and low molecular weight (LMW) particles are shown in Table 3. [T0=time 0; 4W=4 weeks and 8W=8 weeks]

As shown in FIG. 1, fig. 2 and fig. 3, UP-SEC analysis of samples to determine percentage HMW and percentage monomer showed that at 5°C, 25°C and 40°C, all formulations showed a trend towards an increase in peak % HMW and peak % LMW (and a subsequent decrease in peak % monomer) until the 8 week time point. At 25°C, all formulations showed similar trends but smaller changes compared to 40°C. At a temperature of 5°C no significant changes were observed.

As shown in FIG. 2 and 3, formulation 1 shows a greater increase in %HMW and %LMW compared to the other formulations. In addition, formulation 1 showed a greater reduction in % monomer. Thus, all four sets of excipients reduce the %HMW and %LMW and thus improve the stability of the RSV antibody DP.

EXAMPLE 3

Screening of anti-RSV antibody formulation excipients (amino acids and mixtures thereof) to improve the chemical stability of the anti-RSV antibody

This study compared the stability of anti-RSV antibody in 10 mM histidine (pH=6), 0.02% PS80, 7% sucrose in the presence of (i) 50 mM L-Arg.HCl or (ii) L -Lys.HCl at a concentration of 50 mM or (iii) a mixture of L-Arg at a concentration of 25 mM and L-Glu at a concentration of 25 mM or (iv) a mixture of L-Arg.HCl at a concentration of 25 mM and Lys.HCl at a concentration of 25 mM .

These four excipients were examined for improved chemical stability. The four excipients studied were:

(i) L-Arg.HCl at a concentration of 50 mM;

(ii) L-Lys.HCl at a concentration of 50 mM;

(iii) a mixture of L-Arg at a concentration of 25 mM and L-Glu at a concentration of 25 mM; and

(iv) a mixture of L-Arg.HCl at a concentration of 25 mM and Lys.HCl at a concentration of 25 mM.

The first two excipients were studied to understand the effects of positively charged lysine and arginine in the presence of the chloride counteranion. A third excipient was studied to understand the effect of arginine in the presence of another counteranion (glutamate). The fourth excipient was a mixture of the first two excipients to determine whether the two excipients could play a complementary role. It has been suggested that this mixture may play a better role than individual excipients, since the mixture of excipients has different pKa and protects the surface alteration distribution of the monoclonal antibody differently, and this may affect the chemical degradation of the monoclonal antibody.

To evaluate the stability of the formulations, five formulations were bottled and stabilized at 5°C (ambient humidity), 25°C (60% relative humidity) and 40°C (75% relative humidity) for eight weeks, protected from light. place. These five formulations are shown in Example 2.

Ion exchange chromatography was performed to assess chemical stability and monitor changes in the charge variant profile over time. The ion exchange HPLC method was carried out using a Dionex ProPac WCX-10 column and a UV detector at 280 nm. Samples were diluted with purified water and injected at 80 μg for analysis. The mobile phase used for IEX analysis of thermal stability samples was a gradient of the following mobile phases (mobile phase A: 20 mM MOPS, pH 7.2; mobile phase B: 50 mM sodium phosphate, 60 mM sodium chloride, pH 8.0) . The assay is performed using a mobile phase gradient from 20 mM MOPS, pH 7.2 to 50 mM sodium phosphate, 60 mM NaCl, pH 8.0. UV detection was performed at 280 nm.

HP-IEX data for assessing levels of acidic variants, % peak analyte and % alkaline variants are shown in Table 4. [T0=time 0; 4W=4 weeks and 8W=8 weeks]

As shown in FIG. 4, fig. 5 and fig. 6, HP-IEX analysis of samples to determine chemical stability showed that at 5°C, 25°C and 40°C, all formulations showed a trend towards an increase in acidic peak % and a subsequent decrease in monomer peak % until the 8 week time point. At 25°C, all formulations showed similar trends but smaller changes compared to 40°C. At a temperature of 5°C no significant changes were observed.

As can be seen in FIG. 5 and 6, formulation 1 shows a greater increase in acid peak % compared to the other formulations. In addition, composition 1 showed a greater decrease in % peak of the determined component compared to other compositions. Thus, all four sets of excipients reduce the peak % of acidic forms and thus improve the chemical stability of the RSV antibody DP.

EXAMPLE 4

Screening of anti-RSV antibody formulation excipients (to reduce the viscosity of the anti-RSV antibody)

This study compared the stability of anti-RSV antibody in 10 mM histidine (pH=6), 0.02% PS80, 7% sucrose in the presence of (i) 50 mM L-Arg.HCl or (ii) ) L-Lys.HCl at a concentration of 50 mM or (iii) a mixture of L-Arg at a concentration of 25 mM and L-Glu at a concentration of 25 mM or (iv) a mixture of L-Arg.HCl at a concentration of 25 mM and L-Lys.HCl at a concentration of 25 mM.

These four excipients were tested for their ability to reduce the viscosity of the RSV antibody. The four excipients studied were:

(i) L-Arg.HCl at a concentration of 50 mM;

(ii) L-Lys.HCl at a concentration of 50 mM;

(iii) a mixture of L-Arg at a concentration of 25 mM and L-Glu at a concentration of 25 mM; and

(iv) a mixture of L-Arg.HCl at a concentration of 25 mM and L-Lys.HCl at a concentration of 25 mM.

The first two excipients were studied to understand the effects of positively charged lysine and arginine in the presence of the chloride counteranion. A third excipient was studied to understand the effect of arginine in the presence of another counteranion (glutamate). The fourth excipient was a mixture of the first two excipients to see if the two excipients could play a complementary role. It was hypothesized that this mixture may play a better role than individual excipients because the mixture of excipients has different pKa and differentially screens the distribution of surface changes of the monoclonal antibody, thereby increasing the repulsive protein-protein interaction and decreasing the viscosity of the monoclonal antibody.

For viscosity measurements, samples were loaded into a 500 μL syringe and viscosity was measured (five times for each sample) at 20°C using an MVROC viscometer. VROC sensors determine viscosity by measuring the pressure drop as a sample passes through the sensor flow path at specific locations from the inlet. Viscosity was measured as a function of shear rate, with the shear rates of the samples determined from the viscosity of the sample.

As shown in Table 5 and FIG. 7, all four sets of excipients reduce the viscosity of the RSV antibody DP.

EXAMPLE 5

Addition of a chelator to an anti-RSV antibody formulation to reduce aggregation This study compared the stability of an anti-RSV antibody in 10 mM histidine (pH=6), 0.02% PS80, 7% sucrose in the presence or absence of DTPA in concentration of 50 μM (diethylenetriamine pentaacetate).

To evaluate the stability of the formulations, two formulations were bottled and stabilized at 5°C (ambient humidity), 25°C (60% relative humidity) and 40°C (75% relative humidity) for eight weeks, protected from light. place. These two compositions are shown in Table 6.

Samples were assessed by size exclusion chromatography (SEC) for purity, and the percentage of monomer, as well as the percentage of high molecular weight particles (HMW) and late elution peaks (LMW particles), were determined. Ultra-high performance exclusion chromatography (UP-SEC) was performed by diluting samples to 1.0 mg/mL in the mobile phase (100 mM sodium phosphate and 100 mM sodium chloride, pH 7.0); flow rate 0.5 ml/min. Diluted samples were injected (5 μL) into a UPLC equipped with a Waters BEN200 column and UV detector. Proteins in the sample were separated by size and detected by UV absorbance at 214 nm.

UPSEC data for estimating levels of high molecular weight particles (HMW or aggregates), % monomer and LMW (low molecular weight particles) are shown in Table 7. [T0=time 0; 4W=4 weeks and 8W=8 weeks]

As shown in FIG. 8, fig. 9 and fig. 10, UP-SEC analysis of samples to determine % HMW and % monomer showed that at 5°C, 25°C and 40°C, both formulations showed a trend towards an increase in % HMW peak and % LMW peak (and a subsequent decrease in % LMW peak). monomer peak) to the 8 week time point. At 25°C, both formulations showed similar trends but smaller changes compared to 40°C. At a temperature of 5°C no significant changes were observed.

As can be seen in FIG. 9 and 10, formulation 1 showed a greater increase in %HMW and %LMW compared to formulation 2. Additionally, formulation 1 showed a greater decrease in %monomer compared to formulation 2. Thus, DTPA (formulation 2) reduced %HMW and %LMW and thus improves the aggregation of anti-RSV antibody DP.

EXAMPLE 6

Adding a chelator to an anti-RSV antibody formulation to improve the chemical stability of the anti-RSV antibody

This study compares the chemical stability of anti-RSV antibody in 10 mM histidine (pH=6), 0.02% PS80, 7% sucrose in the presence or absence of 50 μM DTPA.

To evaluate the chemical stability of the formulations, two formulations were bottled and assessed for stability at 5°C (ambient humidity), 25°C (60% relative humidity), and 40°C (75% relative humidity) for eight weeks in a protected environment. from the light place. These two compositions are presented in example 5.

Ion exchange chromatography was performed to assess chemical stability and monitor changes in the charge variant profile over time. The ion exchange HPLC method was carried out using a Dionex ProPac WCX-10 column and a UV detector at 280 nm. Samples were diluted with purified water and injected at 80 μg for analysis. The mobile phase used for IEX analysis of thermal stability samples was a gradient of the following mobile phases (mobile phase A: 20 mM MOPS, pH 7.2; mobile phase B: 50 mM sodium phosphate, 60 mM sodium chloride, pH 8.0) . The assay was performed using a mobile phase gradient from 20 mM MOPS, pH 7.2 to 50 mM sodium phosphate, 60 mM NaCl, pH 8.0. UV detection was performed at 280 nm.

HP-IEX data for assessing levels of acidic variants, % peak analyte and % alkaline variants are shown in Table 8. [T0=time 0; 4W=4 weeks and 8W=8 weeks]

As shown in FIG. 11, fig. 12 and fig. 13, HP-IEX analysis of the samples to determine chemical stability showed that at 5°C, 25°C and 40°C, both formulations showed a trend towards an increase in peak % acidic species and a subsequent decrease in peak % monomer) up to the 8 week time point. At 25°C, both formulations showed similar trends but smaller changes compared to 40°C. At a temperature of 5°C no significant changes were observed.

As can be seen in FIG. 12 and 13, formulation 1 showed a greater increase in acid peak % compared to formulation 2. Additionally, formulation 1 showed a greater decrease in analyte peak % compared to formulation 2. Thus, DTPA (formulation 2) reduced acid peak % and thus improves the chemical stability of the RSV antibody DP.

EXAMPLE 7

Adding a chelator to an anti-RSV antibody formulation to reduce PS80 degradation

This study compared the chemical stability of PS80 in anti-RSV antibody in 10 mM histidine (pH=6), 0.02% PS80, 7% sucrose in the presence or absence of 50 μM DTPA. Degradation of PS80 is typically observed during long-term storage of a monoclonal antibody, and an experiment was conducted to determine the compatibility of DTPA with PS80.

To evaluate the chemical stability of PS80 in two formulations, the two formulations were bottled and stabilized at 5°C (ambient humidity), 25°C (60% RH), and 40°C (75% RH) for eight weeks in a dark place. These two compositions are presented in example 5.

The results in Table 9 show similar PS80 degradation profiles for both formulations. In fig. Figure 14 shows the % degradation of PS80 at 40°C. The percentage of degradation was slightly less for formulation 2 (in the presence of DTPA). Thus, DTPA may protect against PS80 degradation, and the two excipients (DTPA and PS80) are mutually compatible. [T0=time 0; 4W=4 weeks and 8W=8 weeks]

EXAMPLE 8

Increasing concentration of excipients to reduce viscosity and maintain stability

To evaluate the effect of increased concentrations of excipients on reducing viscosity, antibodies to RSV were prepared at two different concentrations (about 200 mg/ml and 154 mg/ml) in four different formulations: the first composition containing histidine at a concentration of about 10 mM, sucrose at a concentration 7% and PS80 at a concentration of 0.02%, pH 6, and three formulations with the addition of an additional high concentration of excipients: 70 mM histidine (for a total concentration of 80 mM histidine), 70 mM lysine or 70 mM Arg.

To measure the viscosity of the four formulations, samples were loaded into a 500 μL syringe and the viscosity was measured (four times for each sample) at room temperature using an MVROC viscometer. VROC sensors determine viscosity by measuring the pressure drop as a sample passes through the sensor flow path at specific locations from the inlet. Viscosity was measured as a function of shear rate (measured in reciprocal seconds [s ^-1 ]). The shear rate used was 997 s ^-1 . In fig. 15 shows a bar graph comparing four different anti-RSV antibody formulations at their respective concentrations (about 200 mg/ml and 154 mg/ml).

Compared to a formulation containing about 10 mM histidine, 7% sucrose and 0.02% PS80, pH 6, the addition of higher concentrations of histidine, lysine and arginine showed a significant reduction in viscosity. As shown in FIG. 15, the four formulations did not show significantly different viscosities at an anti-RSV antibody concentration of 154 mg/mL. However, at higher anti-RSV antibody concentrations of approximately 200 mg/mL, formulations using additional 70 mM histidine, 70 mM lysine, or 70 mM arginine showed a significant reduction in mean viscosity compared to the former formulation. Antibody to RSV at a concentration of 213 mg/ml, composed of histidine at a concentration of about 10 mM, sucrose at a concentration of 7% and PS80 at a concentration of 0.02%, pH 6, has an average viscosity of 64.97 centipoise (cP). The use of an additional 70 mM histidine excipient (total histidine concentration 80 mM) reduced the average viscosity of the 204 mg/mL anti-RSV antibody formulation to 30.85 cP. The use of 70 mM lysine as an excipient reduced the average viscosity of the 192 mg/ml anti-RSV antibody formulation to 19.81 cP. The use of arginine at a concentration of 70 mM as an excipient reduced the average viscosity of the 217 mg/ml anti-RSV antibody formulation to 29.27 cP.

The stability of each formulation was monitored for 3 months at temperatures of 5°C, 25°C and 40°C. Samples were assessed by size exclusion chromatography (SEC) for purity, which determined the percentage of monomer, as well as the percentage of high molecular weight particles (HMW) and late elution peaks (LMW particles). Ion exchange chromatography was performed as described in Example 6 to assess chemical stability and monitor changes in the charge variant profile over time. The values are listed in Table 10 below.

Compared to a formulation containing about 10 mM histidine, 7% sucrose, and 0.02% PS80, pH 6, the addition of higher concentrations of histidine, lysine, and arginine did not significantly increase the number of high or low molecular weight species and did not lead to a significant decrease in the percentage of monomer.

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LIST OF SEQUENCES

<110> MERCK SHARP & DOOME CORP.

Hashemi, Venus

De, Arnab

Narasimhan, Chakravarthy Nachu

<120> COMPOSITION OF ANTIBODIES TO RSV AND METHODS OF THEIR APPLICATION

<130> 24641-WO-PCT

<150>US 62/747,418

<151> 2018-10-18

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<170> Patent In version 3.5

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Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Asn Ile

65 70 75 80

Ala Tyr Leu Gln Met Asn Ser Leu Lys Thr Glu Asp Thr Ala Val Tyr

85 90 95

Tyr Cys Thr Arg Gly Ala Pro Tyr Gly Gly Asn Ser Asp Tyr Tyr Tyr

100 105 110

Gly Leu Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala

115 120 125

Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser

130 135 140

Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe

145 150 155 160

Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly

165 170 175

Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu

180 185 190

Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr

195 200 205

Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys

210 215 220

Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro

225 230 235 240

Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys

245 250 255

Pro Lys Asp Thr Leu Tyr Ile Thr Arg Glu Pro Glu Val Thr Cys Val

260 265 270

Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr

275 280 285

Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu

290 295 300

Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His

305 310 315 320

Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys

325 330 335

Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln

340 345 350

Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu

355 360 365

Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro

370 375 380

Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn

385 390 395 400

Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu

405 410 415

Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val

420 425 430

Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln

435 440 445

Lys Ser Leu Ser Leu Ser Pro Gly Lys

450 455

<210> 10

<211> 214

<212> Protein

<213> Artificial sequence

<220>

<223> Light chain RB1+YTE

<400> 10

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Thr Ser Gln Asp Val Arg Gly Ala

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile

35 40 45

Phe Asp Ala Ser Ser Leu Glu Thr Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Val Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Ala Tyr Tyr Cys Gln Gln Phe Leu Asp Phe Pro Phe

85 90 95

Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys Arg Thr Val Ala Ala

100 105 110

Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly

115 120 125

Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala

130 135 140

Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln

145 150 155 160

Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser

165 170 175

Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr

180 185 190

Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser

195 200 205

Phe Asn Arg Gly Glu Cys

210

<---

Claims (25)

1. An anti-RSV antibody composition for treating or preventing RSV infection in a human patient, comprising: a) from about 50 mg/ml to about 250 mg/ml of an antibody to RSV or an antigen binding fragment thereof containing complementarity determining regions (CDRs) having the amino acid sequences: SEQ ID NO: 1 (heavy chain CDR 1), SEQ ID NO: 2 (heavy chain CDR 2), SEQ ID NO: 3 (heavy chain CDR 3), SEQ ID NO: 4 (light chain CDR 1), SEQ ID NO: 5 (light chain CDR 2) and SEQ ID NO: 6 ( CDR 3 light chain); b) about 5 mM to about 20 mM histidine; c) about 4% to about 8% w/v sucrose; d) about 70 mM L-arginine or a pharmaceutically acceptable L-arginine salt; And e) about 0.01% to about 0.10% w/v polysorbate 80. 2. The anti-RSV antibody composition of claim 1, comprising from about 125 mg/ml to about 175 mg/ml of anti-RSV antibody or antigen-binding fragment thereof. 3. An anti-RSV antibody composition according to claim 1, containing about 150 mg/ml of an anti-RSV antibody or an antigen-binding fragment thereof. 4. The anti-RSV antibody composition of claim 1, comprising from about 150 mg/ml to about 200 mg/ml of anti-RSV antibody or antigen-binding fragment thereof. 5. The anti-RSV antibody composition of claim 1, comprising about 175 mg/ml anti-RSV antibody or antigen-binding fragment thereof. 6. The anti-RSV antibody composition of claim 1, comprising from about 175 mg/ml to about 225 mg/ml of anti-RSV antibody or antigen-binding fragment thereof. 7. An anti-RSV antibody composition according to claim 1, containing about 200 mg/ml of an anti-RSV antibody or an antigen-binding fragment thereof. 8. Composition of an antibody to RSV according to any one of paragraphs. 1-7, further containing from about 25 to about 75 μM diethylenetriamine pentaacetate (DTPA). 9. Composition of an antibody to RSV according to any one of paragraphs. 1-8, further containing from about 1 mm to about 20 mm L-methionine. 10. Composition of an antibody to RSV according to any one of paragraphs. 1-9, in which the pH value is from 5.5 to 6.5. 11. Composition of an antibody to RSV according to any one of claims. 1-10, in which the pH value is 6.0. 12. Composition of an antibody to RSV according to any one of claims. 1-11, wherein the anti-RSV antibody comprises a heavy chain variable region having the amino acid sequence of SEQ ID NO: 7 and a light chain variable region having the amino acid sequence. SEQ ID NO: 8. 13. Composition of an antibody to RSV according to any one of paragraphs. 1-11, wherein the anti-RSV antibody includes an immunoglobulin heavy chain consisting of the amino acid sequence set forth in SEQ ID NO: 9 and an immunoglobulin light chain consisting of the amino acid sequence set forth in SEQ ID NO: 10. 14. Use of an anti-RSV antibody composition according to any one of claims. 1-13 for the treatment or prevention of RSV infection in a human patient in need thereof. 15. Use of the anti-RSV antibody composition of claim 14, wherein the anti-RSV antibody composition is applied in an effective amount that provides a fixed dose of the anti-RSV antibody or antigen binding fragment thereof from about 10 mg to about 150 mg. 16. Use of the anti-RSV antibody composition of claim 14, wherein the anti-RSV antibody composition is applied in an effective amount that provides a fixed dose of the anti-RSV antibody or antigen binding fragment thereof from about 25 mg to about 125 mg. 17. Use of the anti-RSV antibody composition of claim 14, wherein the anti-RSV antibody composition is applied in an effective amount that provides a fixed dose of the anti-RSV antibody or antigen binding fragment thereof from about 50 mg to about 100 mg. 18. Use of the anti-RSV antibody composition of claim 14, wherein the anti-RSV antibody composition is applied in an effective amount that provides a fixed dose of the anti-RSV antibody or antigen binding fragment thereof of about 100 mg. 19. Use of an anti-RSV antibody composition according to any one of claims. 14-18, in which the anti-RSV antibody formulation is administered intramuscularly. 20. Use of an anti-RSV antibody composition according to any one of claims. 1-13 for the manufacture of a medicament for the treatment or prevention of RSV infection in a human patient.

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