Patent Description:
Therapeutic macromolecules (e.g., antibodies) must be formulated in a manner that not only makes the molecules suitable for administration to patients, but also maintains their stability during storage. For example, therapeutic antibodies in liquid solution are prone to degradation, aggregation and/or undesired chemical modifications unless the solution is formulated properly. The stability of an antibody in liquid formulation depends not only on the kinds of excipients used in the formulation, but also on the amounts and proportions of the excipients relative to one another. Furthermore, other considerations aside from stability must be taken into account when preparing a liquid antibody formulation. Examples of such additional considerations include the viscosity of the solution and the concentration of antibody that can be accommodated by a given formulation. Thus, when formulating a therapeutic antibody, great care must be taken to arrive at a formulation that remains stable, contains an adequate concentration of antibody, and possesses a suitable viscosity as well as other properties which enable the formulation to be conveniently administered to patients.

Antibodies to the human interleukin-<NUM> receptor (hIL-6R) are one example of a therapeutically relevant macromolecule that requires proper formulation. Anti-hIL-6R antibodies are clinically useful for the treatment and/or prevention of diseases such as rheumatoid arthritis, ankylosing spondylitis, and other conditions. Exemplary anti-IL-6R antibodies are described, inter alia, in <CIT>; <CIT>; <CIT>; <CIT>; and <CIT>. A particularly important anti-hIL-6R antibody with great therapeutic potential is the antibody referred to in <CIT> as VQ8F11-<NUM> (also referred to herein as "mAb1").

Although anti-hIL-6R antibodies are known, there remains a need in the art for novel pharmaceutical formulations comprising anti-hIL-6R antibodies which are sufficiently stable and also suitable for administration to patients.

The present invention provides a pharmaceutical formulation for use in a method of treatment, prevention and/or amelioration of any disease or disorder associated with IL-<NUM> activity, wherein the method comprises injection of the pharmaceutical formulation into the subject, and wherein the pharmaceutical formulation comprises:.

Also described herein are pharmaceutical formulations comprising a human antibody that specifically binds to human interleukin-<NUM> receptor (hIL-6R). The formulations of the disclosure may comprise excipients in addition to the anti-hIL-6R antibody. For example, in certain aspects, the formulation may comprise (i) a human antibody that specifically binds to hIL-6R; (ii) at least one amino acid; and (iii) at least one carbohydrate. The amino acid can be, e.g., histidine and/or arginine. The carbohydrate can be a sugar such as, e.g., sucrose, glucose, mannitol, lactose or trehalose.

According to certain aspects of the present disclosure, the formulation further comprises a non-ionic surfactant. The non-ionic surfactant can be, e.g., polysorbate <NUM>, polysorbate <NUM>, polyoxyethylene sorbitan monooleate, polyethylene glycol, etc..

The antibody contained within the pharmaceutical formulations of the present disclosure can be any antibody which specifically binds to hIL-6R. Exemplary antibodies that may be contained within the formulations of the disclosure are antibodies comprising a heavy chain variable region (HCVR) and a light chain variable region (LCVR), wherein the HCVR comprises a heavy chain complementary determining region (HCDR) <NUM> having the amino acid sequence of SEQ ID NO: <NUM>, a HCDR2 having the amino acid sequence of SEQ ID NO:<NUM>, and a HCDR3 having the amino acid sequence of SEQ ID NO:<NUM>; and wherein the LCVR comprises a light chain complementary determining region (LCDR) <NUM> having the amino acid sequence of SEQ ID NO: <NUM>, a LCDR2 having the amino acid sequence of SEQ ID NO:<NUM>, and a LCDR3 having the amino acid sequence of SEQ ID NO:<NUM>. The antibody contained within the formulations of the present invention are antibodies comprising a HCVR having the amino acid sequence of SEQ ID NO:<NUM> and a LCVR having the amino acid sequence of SEQ ID NO:<NUM>.

The antibody formulations of the present invention may be contained within any suitable container useful for storing pharmaceutical formulations. Examples of such suitable containers include, e.g., glass or plastic vials, syringes and cartridges. The container may be clear or opaque (e.g., amber colored).

According to certain aspects, the pharmaceutical formulations remain relatively stable following storage for several days, months or years at a given temperature. For example, in certain exemplary aspects, a high percentage of the antibody (e.g., <NUM>%, <NUM>%, <NUM>% or more) is maintained in its native form following at least <NUM>, <NUM>, <NUM> or more months of storage. The percentage of native form of the antibody may be measured, e.g., by SE-HPLC, or by any other method known in the art. The storage temperature at which stability of the antibody is maintained can be, e.g., -<NUM>, -<NUM>, -<NUM>, <NUM>, <NUM>, <NUM>, <NUM>, or higher.

Before the present invention is described, it is to be understood that this invention is not limited to particular methods and experimental conditions described, as such methods and conditions may vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting, since the scope of the present invention will be limited only by the appended claims.

As used herein, the term "about," when used in reference to a particular recited numerical value, means that the value may vary from the recited value by no more than <NUM>%. For example, as used herein, the expression "about <NUM>" includes <NUM> and <NUM> and all values in between (e.g., <NUM>, <NUM>, <NUM>, <NUM>, etc.).

Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, the preferred methods and materials are now described.

As used herein, the expression "pharmaceutical formulation" means a combination of at least one active ingredient (e.g., a small molecule, macromolecule, compound, etc. which is capable of exerting a biological effect in a human or non-human animal), and at least one inactive ingredient which, when combined with the active ingredient and/or one or more additional inactive ingredients, is suitable for therapeutic administration to a human or non-human animal. The term "formulation," as used herein, means "pharmaceutical formulation" unless specifically indicated otherwise. The present disclosure provides pharmaceutical formulations comprising at least one therapeutic polypeptide. According to certain aspects of the present disclosure, the therapeutic polypeptide is an antibody that binds specifically to human interleukin-<NUM> receptor (hIL-6R) or an antigen-binding fragment thereof. More specifically, the present disclosure includes pharmaceutical formulations that comprise: (i) a human antibody that specifically binds to hIL-6R; (ii) histidine; and (iii) a carbohydrate. Additional components may be included in the formulations of the present disclosure such as, e.g., at least one non-ionic surfactant, and at least one additional amino acid. Specific exemplary components and formulations included within the present disclosure are described in detail below.

The pharmaceutical formulations may, in certain aspects, be fluid formulations. As used herein, the expression "fluid formulation" means a mixture of at least two components that exists predominantly in the fluid state at about <NUM> to about <NUM>. Fluid formulations include, inter alia, liquid formulations. Fluid formulations may be of low, moderate or high viscosity depending on their particular constituents.

Specifically, the present invention provides a pharmaceutical formulation for use in a method of treatment, prevention and/or amelioration of any disease or disorder associated with IL-<NUM> activity, wherein the method comprises injection of the pharmaceutical formulation into the subject, and wherein the pharmaceutical formulation comprises:.

The pharmaceutical formulations of the present disclosure may comprise a human antibody, or an antigen-binding fragment thereof, that binds specifically to hIL-6R. As used herein, the term "hIL-6R" means a human cytokine receptor that specifically binds interleukin-<NUM> (IL-<NUM>). In certain aspects, the antibody contained within the pharmaceutical formulations of the present disclosure binds specifically to the extracellular domain of hIL-6R. The extracellular domain of hIL-6R is represented by the amino acid sequence of SEQ ID NO:<NUM>. In the present invention, the pharmaceutical formulation comprises a human antibody as set forth in the claims.

The term "antibody", as used herein, is generally intended to refer to immunoglobulin molecules comprising four polypeptide chains, two heavy (H) chains and two light (L) chains inter-connected by disulfide bonds, as well as multimers thereof (e.g., IgM); however, immunoglobulin molecules consisting of only heavy chains (i.e., lacking light chains) are also encompassed within the definition of the term "antibody. " Each heavy chain comprises a heavy chain variable region (abbreviated herein as HCVR or VH) and a heavy chain constant region. The heavy chain constant region comprises three domains, CH1, CH2 and CH3. Each light chain comprises a light chain variable region (abbreviated herein as LCVR or VL) and a light chain constant region. The light chain constant region comprises one domain (CL1). The VH and VL regions can be further subdivided into regions of hypervariability, termed complementary determining regions (CDRs), interspersed with regions that are more conserved, termed framework regions (FR). Each VH and VL is composed of three CDRs and four FRs, arranged from amino-terminus to carboxy-terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4.

Unless specifically indicated otherwise, the term "antibody," as used herein, shall be understood to encompass complete antibody molecules as well as antigen-binding fragments thereof. The term "antigen-binding portion" or "antigen-binding fragment" of an antibody (or simply "antibody portion" or "antibody fragment"), as used herein, refers to one or more fragments of an antibody that retain the ability to specifically bind to hIL-6R.

An "isolated antibody", as used herein, is intended to refer to an antibody that is substantially free of other antibodies having different antigenic specificities (e.g., an isolated antibody that specifically binds hIL-6R is substantially free of antibodies that specifically bind antigens other than hlL-6R).

The term "specifically binds," or the like, means that an antibody or antigen-binding fragment thereof forms a complex with an antigen that is relatively stable under physiologic conditions. Specific binding can be characterized by a dissociation constant of at least about 1x10-<NUM> M or greater. Methods for determining whether two molecules specifically bind are well known in the art and include, for example, equilibrium dialysis, surface plasmon resonance, and the like. An isolated antibody that specifically binds hIL-6R may, however, have cross-reactivity to other antigens, such as IL-6R molecules from other species. In the context of the present disclosure, multispecific (e.g., bispecific) antibodies that bind to hIL-6R as well as one or more additional antigens are deemed to "specifically bind" hIL-6R. Moreover, an isolated antibody may be substantially free of other cellular material and/or chemicals.

Exemplary anti-hIL-6R antibodies that may be included in the pharmaceutical formulations described herein are set forth in <CIT>.

According to certain aspects of the present disclosure, the anti-hIL-6R antibody, or antigen-binding fragment thereof, comprises a heavy chain complementary determining region (HCDR) <NUM> having an amino acid sequence selected from the group consisting of SEQ ID NO: <NUM>, <NUM>, <NUM> and <NUM>; a HCDR2 having an amino acid sequence selected from the group consisting of SEQ ID NO:<NUM>, <NUM>, <NUM> and <NUM>; and a HCDR3 having an amino acid sequence selected from the group consisting of SEQ ID NO:<NUM>, <NUM>, <NUM> and <NUM>. In certain aspects, the anti-hIL-6R antibody, or antigen-binding fragment thereof, comprises HCDR1-HCDR2-HCDR3 domains, respectively, selected from the group consisting of: (i) SEQ ID NOs:<NUM> - <NUM> - <NUM>; (ii) SEQ ID NOs:<NUM> - <NUM> - <NUM>; (iii) SEQ ID NOs:<NUM> - <NUM> - <NUM>; and (iv) SEQ ID NOs:<NUM> - <NUM> - <NUM>.

According to certain aspects of the present disclosure, the anti-hIL-6R antibody, or antigen-binding fragment thereof, comprises a light chain complementary determining region (LCDR) <NUM> having an amino acid sequence selected from the group consisting of SEQ ID NO: <NUM>, <NUM>, <NUM> and <NUM>; a LCDR2 having an amino acid sequence selected from the group consisting of SEQ ID NO:<NUM>, <NUM>, <NUM> and <NUM>; and a LCDR3 having an amino acid sequence selected from the group consisting of SEQ ID NO:<NUM>, <NUM>, <NUM> and <NUM>. In certain aspects, the anti-hIL-6R antibody, or antigen-binding fragment thereof, comprises LCDR1-LCDR2-LCDR3 domains, respectively, selected from the group consisting of: (i) SEQ ID NOs:<NUM> - <NUM> - <NUM>; (ii) SEQ ID NOs:<NUM> - <NUM> - <NUM>; (iii) SEQ ID NOs:<NUM> - <NUM> - <NUM>; and (iv) SEQ ID NOs:<NUM> - <NUM> - <NUM>.

In certain aspects, the anti-hIL-6R antibody, or antigen-binding fragment thereof, comprises HCDR1-HCDR2-HCDR3 / LCDR1-LCDR2-LCDR3 domains, respectively, selected from the group consisting of: (i) SEQ ID NOs:<NUM> - <NUM> - <NUM> / SEQ ID NOs:<NUM> - <NUM> - <NUM>; (ii) SEQ ID NOs:<NUM> - <NUM> - <NUM> / SEQ ID NOs:<NUM> - <NUM> - <NUM>; (iii) SEQ ID NOs:<NUM> - <NUM> - <NUM> / SEQ ID NOs:<NUM> - <NUM> - <NUM>; and (iv) SEQ ID NOs:<NUM> - <NUM> - <NUM> / SEQ ID NOs:<NUM> - <NUM> - <NUM>.

In certain aspects, the anti-hIL-6R antibody, or antigen-binding fragment thereof, comprises a heavy chain variable region (HCVR) having an amino acid sequence selected from the group consisting of SEQ ID NO:<NUM>, <NUM>, <NUM> and <NUM>. In certain aspects, the anti-hIL-6R antibody, or antigen-binding fragment thereof, comprises a light chain variable region (LCVR) having an amino acid sequence selected from the group consisting of SEQ ID NO:<NUM>, <NUM>, <NUM> and <NUM>. In certain aspects, the anti-hIL-6R antibody, or antigen-binding fragment thereof, comprises a HCVR/LCVR amino acid sequence pair selected from the group consisting of SEQ ID NO:<NUM>/<NUM>; <NUM>/<NUM>; <NUM>/<NUM> and <NUM>/<NUM>.

The non-limiting, exemplary antibody used in the Examples herein is referred to as "mAb1. " This antibody is also referred to in <CIT> as VQ8F11-<NUM>. mAb1 (VQ8F11-<NUM>) comprises an HCVR/LCVR amino acid sequence pair having SEQ ID NOs:<NUM>/<NUM>, and HCDR1-HCDR2-HCDR3 / LCDR1-LCDR2-LCDR3 domains represented by SEQ ID NOs:<NUM> - <NUM> - <NUM> / SEQ ID NOs:<NUM> - <NUM> - <NUM>.

The amount of antibody, or antigen-binding fragment thereof, contained within the pharmaceutical formulations of the present disclosure may vary depending on the specific properties desired of the formulations, as well as the particular circumstances and purposes for which the formulations are intended to be used. The pharmaceutical formulations may contain about <NUM>/mL to about <NUM>/mL of antibody; about <NUM>/mL to about <NUM>/mL of antibody; about <NUM>/mL to about <NUM>/mL of antibody; about <NUM>/mL to about <NUM>/mL of antibody; about <NUM>/mL to about <NUM>/mL of antibody; or about <NUM>/mL to about <NUM>/mL of antibody. For example, the formulations may comprise about <NUM>/mL; about <NUM>/mL; about <NUM>/mL; about <NUM>/mL; about <NUM>/mL; about <NUM>/mL; about <NUM>/mL; about <NUM>/mL; about <NUM>/mL; about <NUM>/mL; about <NUM>/mL; about <NUM>/mL; about <NUM>/mL; about <NUM>/mL; about <NUM>/mL; about <NUM>/mL; about <NUM>/mL; about <NUM>/mL; about <NUM>/mL; about <NUM>/mL; about <NUM>/mL; about <NUM>/mL; about <NUM>/mL; about <NUM>/mL; about <NUM>/mL; about <NUM>/mL; about <NUM>/mL; about <NUM>/mL; about <NUM>/mL; about <NUM>/mL; about <NUM>/mL; about <NUM>/mL; about <NUM>/mL; about <NUM>/mL; about <NUM>/mL; about <NUM>/mL; about <NUM>/mL; about <NUM>/mL; about <NUM>/mL; about <NUM>/mL; about <NUM>/mL; about <NUM>/mL; about <NUM>/mL; about <NUM>/mL; about <NUM>/mL; about <NUM>/mL; about <NUM>/mL; about <NUM>/mL; about <NUM>/mL; or about <NUM>/mL of an antibody or an antigen-binding fragment thereof, that binds specifically to hIL-6R.

The pharmaceutical formulations of the present disclosure comprise one or more excipients. The term "excipient," as used herein, means any non-therapeutic agent added to the formulation to provide a desired consistency, viscosity or stabilizing effect.

In certain aspects, the pharmaceutical formulation of the disclosure comprises at least one amino acid. Exemplary amino acids suitable for use in the formulations of the present disclosure include, inter alia, arginine and/or histidine. In the present invention the pharmaceutical formulation comprises histidine and arginine.

The amount of amino acid contained within the pharmaceutical formulations of the present disclosure may vary depending on the specific properties desired of the formulations, as well as the particular circumstances and purposes for which the formulations are intended to be used. The formulations may contain about <NUM> to about <NUM> of an amino acid; about <NUM> to about <NUM> of an amino acid; about <NUM> to about <NUM> of an amino acid; or about <NUM> to about <NUM> of an amino acid. For example, the pharmaceutical formulations of the present disclosure may comprise about <NUM>; about <NUM>; about <NUM>; about <NUM>; about <NUM>; about <NUM>; about <NUM>; about <NUM>; about <NUM>; about <NUM>; about <NUM>; about <NUM>; about <NUM>; about <NUM>; about <NUM>; about <NUM>; about <NUM>; about <NUM>; about <NUM>; about <NUM>; about <NUM>; about <NUM>; about <NUM>; about <NUM>; about <NUM>; about <NUM>; about <NUM>; about <NUM>; about <NUM>; about <NUM>; <NUM>; about <NUM>; about <NUM>; about <NUM>; about <NUM>; about <NUM>; about <NUM>; about <NUM>; about <NUM>; about <NUM>; about <NUM>; about <NUM>; about <NUM>; about <NUM>; about <NUM>; about <NUM>; about <NUM>; about <NUM>; about <NUM>; about <NUM>; about <NUM>; about <NUM>; about <NUM>; about <NUM>; about <NUM>; about <NUM>; about <NUM>; about <NUM>; about <NUM>; about <NUM>; about <NUM>; about <NUM>; or about <NUM> of an amino acid (e.g., histidine and/or arginine). In the present invention, the pharmaceutical formulation comprises histidine at a concentration of from <NUM> to <NUM> and arginine at a concentration of from <NUM> to <NUM>.

The pharmaceutical formulations of the present disclosure may also comprise one or more carbohydrate, e.g., one or more sugar. The sugar can be a reducing sugar or a non-reducing sugar. "Reducing sugars" include, e.g., sugars with a ketone or aldehyde group and contain a reactive hemiacetal group, which allows the sugar to act as a reducing agent. Specific examples of reducing sugars include fructose, glucose, glyceraldehyde, lactose, arabinose, mannose, xylose, ribose, rhamnose, galactose and maltose. Non-reducing sugars can comprise an anomeric carbon that is an acetal and is not substantially reactive with amino acids or polypeptides to initiate a Maillard reaction. Specific examples of non-reducing sugars include sucrose, trehalose, sorbose, sucralose, melezitose and raffinose. Sugar acids include, for example, saccharic acids, gluconate and other polyhydroxy sugars and salts thereof. In the present invention, the pharmaceutical formulation comprises sucrose.

The amount of sugar contained within the pharmaceutical formulations will vary depending on the specific circumstances and intended purposes for which the formulations are used. The formulations may contain about <NUM>% to about <NUM>% sugar; about <NUM>% to about <NUM>% sugar; about <NUM>% to about <NUM>% sugar; about <NUM>% to about <NUM>% sugar; about <NUM>% to about <NUM>% sugar; about <NUM>% to about <NUM>% sugar; or about <NUM>% to about <NUM>% sugar. For example, the pharmaceutical formulations may comprise about <NUM>%; about <NUM>%; about <NUM>%; about <NUM>%; about <NUM>%; about <NUM>%; about <NUM>%; about <NUM>%; about <NUM>%; about <NUM>%; about <NUM>%; about <NUM>%; <NUM>%; about <NUM>%; about <NUM>%; about <NUM>%; about <NUM>%; about <NUM>%; about <NUM>%; about <NUM>%; about <NUM>%; about <NUM>%; about <NUM>%; about <NUM>%; about <NUM>%; about <NUM>%; about <NUM>%; about <NUM>%; about <NUM>%; about <NUM>%; about <NUM>%; about <NUM>%; <NUM>%; about <NUM>%; about <NUM>%; about <NUM>%; about <NUM>%; about <NUM>%; about <NUM>%; or about <NUM>% sugar (e.g., sucrose). In the present invention, the pharmaceutical formulation comprises sucrose in an amount of from <NUM>% to <NUM>% w/v.

The pharmaceutical formulations may also comprise one or more surfactant. As used herein, the term "surfactant" means a substance which reduces the surface tension of a fluid in which it is dissolved and/or reduces the interfacial tension between oil and water. Surfactants can be ionic or non-ionic. Exemplary non-ionic surfactants that can be included in the formulations of the present disclosure include, e.g., alkyl poly(ethylene oxide), alkyl polyglucosides (e.g., octyl glucoside and decyl maltoside), fatty alcohols such as cetyl alcohol and oleyl alcohol, cocamide MEA, cocamide DEA, and cocamide TEA. Specific non-ionic surfactants that can be included in the formulations of the present disclosure include, e.g., polysorbates such as polysorbate <NUM>, polysorbate <NUM>, polysorbate <NUM>, polysorbate <NUM>, polysorbate <NUM>, polysorbate <NUM>, polysorbate <NUM>, and polysorbate <NUM>; poloxamers such as poloxamer <NUM>, poloxamer <NUM>; polyethylene-polypropylene glycol; or polyethylene glycol (PEG). Polysorbate <NUM> is also known as TWEEN <NUM>, sorbitan monolaurate and polyoxyethylenesorbitan monolaurate. In the present invention, the pharmaceutical formulation comprises polysorbate <NUM>.

The amount of surfactant contained within the pharmaceutical formulations of the present disclosure may vary depending on the specific properties desired of the formulations, as well as the particular circumstances and purposes for which the formulations are intended to be used. The formulations may contain about <NUM>% to about <NUM>% surfactant; or about <NUM>% to about <NUM>% surfactant. For example, the formulations may comprise about <NUM>%; about <NUM>%; about <NUM>%; about <NUM>%; about <NUM>%; about <NUM>%; about <NUM> %; about <NUM>%; about <NUM>%; about <NUM>%; about <NUM>%; about <NUM>%; about <NUM>%; about <NUM>%; about <NUM>%; about <NUM>%; about <NUM>%; about <NUM>%; about <NUM>%; about <NUM>%; about <NUM>%; about <NUM>%; about <NUM>%; about <NUM>%; about <NUM>%; or about <NUM>% surfactant (e.g., polysorbate <NUM>). In the present invention, the pharmaceutical formulation comprises polysorbate <NUM> in an amount of from <NUM>% to <NUM>% w/v.

The pharmaceutical formulations described herein may have a pH of from about <NUM> to about <NUM>. For example, the formulations may have a pH of about <NUM>; about <NUM>; about <NUM>; about <NUM>; about <NUM>; about <NUM>; about <NUM>; about <NUM>; about <NUM>; about <NUM>; about <NUM>; about <NUM>; about <NUM>; about <NUM>; about <NUM>; or about <NUM>.

According to one aspect of the present disclosure, the pharmaceutical formulation comprises: (i) a human antibody that specifically binds to hIL-6R (e.g., mAb1); (ii) an amino acid (e.g., histidine); and (iii) a sugar (e.g., sucrose). Specific, non-limiting exemplary aspects encompassed by this aspect of the disclosure are set forth in Table <NUM>. The formulations in Tables <NUM>, 2A, 2B, 3A, 3C are not in the scope of the claims. The formulations in the first four columns of each of the Tables 3B, 3D, 3E, 3F are not in the scope of the claims.

According to another aspect of the present disclosure, the pharmaceutical formulation comprises: (i) a human antibody that specifically binds to hIL-6R (e.g., mAb1); (ii) an amino acid (e.g., histidine); (iii) a sugar (e.g., sucrose); and (iv) a surfactant (e.g., polysorbate <NUM>). Specific, non-limiting exemplary aspects encompassed by this aspect of the disclosure are set forth in Tables 2A and 2B.

According to another aspect of the present disclosure, the pharmaceutical formulation comprises: (i) a human antibody that specifically binds to hIL-6R (e.g., mAb1); (ii) a first amino acid (e.g., histidine); (iii) a sugar (e.g., sucrose); (iv) a surfactant (e.g., polysorbate <NUM>); and (v) a second amino acid (e.g., arginine). Specific, non-limiting exemplary aspects encompassed by this aspect of the disclosure are set forth in Tables 3A, 3B, 3C, 3D, 3E and 3F.

Additional non-limiting examples of pharmaceutical formulations encompassed by the present disclosure are set forth elsewhere herein, including the working Examples presented below.

The pharmaceutical formulations described herein typically exhibit high levels of stability. The term "stable," as used herein in reference to the pharmaceutical formulations, means that the antibodies within the pharmaceutical formulations retain an acceptable degree of structure and/or function and/or biological activity after storage for a defined amount of time. A formulation may be stable even though the antibody contained therein does not maintain <NUM>% of its structure and/or function and/or biological activity after storage for a defined amount of time. Under certain circumstances, maintenance of about <NUM>%, about <NUM>%, about <NUM>%, about <NUM>%, about <NUM>%, about <NUM>%, about <NUM>% or about <NUM>% of an antibody's structure and/or function and/or biological activity after storage for a defined amount of time may be regarded as "stable.

Stability can be measured, inter alia, by determining the percentage of native antibody remaining in the formulation after storage for a defined amount of time at a given temperature. The percentage of native antibody can be determined by, inter alia, size exclusion chromatography (e.g., size exclusion high performance liquid chromatography [SE-HPLC]). An "acceptable degree of stability," as that phrase is used herein, means that at least <NUM>% of the native form of the antibody can be detected in the formulation after storage for a defined amount of time at a given temperature. In certain aspects, at least about <NUM>%, <NUM>%, <NUM>%, <NUM>%, <NUM>%, <NUM>%, <NUM>%, <NUM>%, <NUM>%, <NUM>% or <NUM>% of the native form of the antibody can be detected in the formulation after storage for a defined amount of time at a given temperature. The defined amount of time after which stability is measured can be at least <NUM> month, at least <NUM> months, at least <NUM> months, at least <NUM> months, at least <NUM> months, at least <NUM> months, at least <NUM> months, at least <NUM> months, at least <NUM> months, at least <NUM> months, at least <NUM> months, at least <NUM> months, at least <NUM> months, at least <NUM> months, or more. The temperature at which the pharmaceutical formulation may be stored when assessing stability can be any temperature from about -<NUM> to about <NUM>, e.g., storage at about -<NUM>, about -<NUM>, about <NUM>, about <NUM>, about <NUM>, or about <NUM>. For example, a pharmaceutical formulation may be deemed stable if after <NUM> months of storage at <NUM>, greater than about <NUM>%, <NUM>%, <NUM>% or <NUM>% of native antibody is detected by SE-HPLC. A pharmaceutical formulation may also be deemed stable if after <NUM> months of storage at <NUM>, greater than about <NUM>%, <NUM>%, <NUM>% or <NUM>% of native antibody is detected by SE-HPLC. A pharmaceutical formulation may also be deemed stable if after <NUM> months of storage at <NUM>, greater than about <NUM>%, <NUM>%, <NUM>% or <NUM>% of native antibody is detected by SE-HPLC. A pharmaceutical formulation may also be deemed stable if after <NUM> months of storage at <NUM>, greater than about <NUM>%, <NUM>%, <NUM>% or <NUM>% of native antibody is detected by SE-HPLC. A pharmaceutical formulation may also be deemed stable if after <NUM> months of storage at <NUM>, greater than about <NUM>%, <NUM>%, <NUM>% or <NUM>% of native antibody is detected by SE-HPLC. A pharmaceutical formulation may also be deemed stable if after <NUM> months of storage at <NUM>, greater than about <NUM>%, <NUM>%, <NUM>% or <NUM>% of native antibody is detected by SE-HPLC.

Other methods may be used to assess the stability of the formulations such as, e.g., differential scanning calorimetry (DSC) to determine thermal stability, controlled agitation to determine mechanical stability, and absorbance at about <NUM> or about <NUM> to determine solution turbidities. For example, a formulation may be considered stable if, after <NUM> or more months of storage at about <NUM> to about <NUM>, the change in OD<NUM> of the formulation is less than about <NUM> (e.g., <NUM>, <NUM>, <NUM>, <NUM>, or less) from the OD<NUM> of the formulation at t=<NUM>.

Stability may also be assessed by measuring the biological activity and/or binding affinity of the antibody to its target. For example, a formulation may be regarded as stable if, after storage at e.g., <NUM>, <NUM>, <NUM>, etc. for a defined amount of time (e.g., <NUM> to <NUM> months), the anti-IL-6R antibody contained within the formulation binds to IL-6R with an affinity that is at least <NUM>%, <NUM>%, <NUM>%, <NUM>%, <NUM>%, <NUM>%, or more of the binding affinity of the antibody prior to said storage. Additional methods for assessing the stability of an antibody in formulation are demonstrated in the Examples presented below.

In the fluid form, the pharmaceutical formulations may, exhibit low to moderate levels of viscosity. "Viscosity" as used herein may be "kinematic viscosity" or "absolute viscosity. " "Kinematic viscosity" is a measure of the resistive flow of a fluid under the influence of gravity. When two fluids of equal volume are placed in identical capillary viscometers and allowed to flow by gravity, a viscous fluid takes longer than a less viscous fluid to flow through the capillary. For example, if one fluid takes <NUM> seconds to complete its flow and another fluid takes <NUM> seconds, the second fluid is twice as viscous as the first on a kinematic viscosity scale. "Absolute viscosity", sometimes called dynamic or simple viscosity, is the product of kinematic viscosity and fluid density (Absolute Viscosity = Kinematic Viscosity x Density). The dimension of kinematic viscosity is L<NUM>/T where L is a length and T is a time. Commonly, kinematic viscosity is expressed in centistokes (cSt). The SI unit of kinematic viscosity is mm<NUM>/s, which is <NUM> cSt. Absolute viscosity is expressed in units of centipoise (cP). The SI unit of absolute viscosity is the milliPascal-second (mPa-s), where <NUM> cP = <NUM> mPa-s.

As used herein, a low level of viscosity, in reference to a fluid formulation of the present disclosure, will exhibit an absolute viscosity of less than about <NUM> cPoise (cP). For example, a fluid formulation of the disclosure will be deemed to have "low viscosity," if, when measured using standard viscosity measurement techniques, the formulation exhibits an absolute viscosity of about <NUM> cP, about <NUM> cP, about <NUM> cP, about <NUM> cP, about <NUM> cP, about <NUM> cP, about <NUM> cP, about <NUM> cP, about <NUM> cP, about <NUM> cP, about <NUM> cP, about <NUM> cP, about <NUM> cP, about <NUM> cP, about <NUM> cP, about <NUM> cP, or less. As used herein, a moderate level of viscosity, in reference to a fluid formulation of the present disclosure, will exhibit an absolute viscosity of between about <NUM> cP and about <NUM> cP. For example, a fluid formulation of the disclosure will be deemed to have "moderate viscosity," if when measured using standard viscosity measurement techniques, the formulation exhibits an absolute viscosity of about <NUM> cP, about <NUM> cP, about <NUM> cP, about <NUM> cP, about <NUM> cP, about <NUM> cP, about <NUM> cP, about <NUM> cP, about <NUM> cP, about <NUM> cP or about <NUM> cP.

As illustrated in Example <NUM> below, the present inventors have made the surprising discovery that low to moderate viscosity fluid formulations comprising high concentrations of an anti-hIL-6R antibody (e.g., up to at least <NUM>/mL) can be obtained by formulating the antibody with <NUM> to <NUM> histidine and <NUM> to <NUM> arginine. In addition, it was further discovered that the viscosity of the formulation could be decreased to an even greater extent by adjusting the sucrose content to less than <NUM>%.

The pharmaceutical formulations disclosed herein may be contained within any container suitable for storage of medicines and other therapeutic compositions. For example, the pharmaceutical formulations may be contained within a sealed and sterilized plastic or glass container having a defined volume such as a vial, ampule, syringe, cartridge, or bottle. Different types of vials can be used to contain the formulations of the present disclosure including, e.g., clear and opaque (e.g., amber) glass or plastic vials. Likewise, any type of syringe can be used to contain and/or administer the pharmaceutical formulations of the present disclosure.

The pharmaceutical formulations described herein may be contained within "normal tungsten" syringes or "low tungsten" syringes. As will be appreciated by persons of ordinary skill in the art, the process of making glass syringes generally involves the use of a hot tungsten rod which functions to pierce the glass thereby creating a hole from which liquids can be drawn and expelled from the syringe. This process results in the deposition of trace amounts of tungsten on the interior surface of the syringe. Subsequent washing and other processing steps can be used to reduce the amount of tungsten in the syringe. As used herein, the term "normal tungsten" means that the syringe contains greater than <NUM> parts per billion (ppb) of tungsten. The term "low tungsten" means that the syringe contains less than <NUM> ppb of tungsten. For example, a low tungsten syringe can contain less than about <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM>, <NUM> or fewer ppb of tungsten.

The rubber plungers used in syringes, and the rubber stoppers used to close the openings of vials, may be coated to prevent contamination of the medicinal contents of the syringe or vial and/or to preserve their stability. Thus, pharmaceutical formulations described herein may be contained within a syringe that comprises a coated plunger, or within a vial that is sealed with a coated rubber stopper. For example, the plunger or stopper may be coated with a fluorocarbon film. Examples of coated stoppers and/or plungers suitable for use with vials and syringes containing the pharmaceutical formulations of the present disclosure are mentioned in, e.g., <CIT>; <CIT>; <CIT>; <CIT>; and <CIT>. Particular exemplary coated rubber stoppers and plungers that can be used in the context of the present disclosure are commercially available under the tradename "FluroTec®," available from West Pharmaceutical Services, Inc. (Lionville, PA).

According to certain aspects of the present invention, the pharmaceutical formulations may be contained within a low tungsten syringe that comprises a fluorocarbon-coated plunger. As discussed in the Examples section below, the combination of a low tungsten syringe and a fluorocarbon-coated plunger was observed to yield surprising stability characteristics with regard to the pharmaceutical formulations described herein.

The pharmaceutical formulations can be administered to a patient by parenteral routes such as injection (e.g., subcutaneous, intravenous, intramuscular, intraperitoneal, etc.) or percutaneous, mucosal, nasal, pulmonary and/or oral administration. Numerous reusable pen and/or autoinjector delivery devices can be used to subcutaneously deliver the pharmaceutical formulations described herein. Examples include, but are not limited to AUTOPEN™ (Owen Mumford, Inc. , Woodstock, UK), DISETRONIC™ pen (Disetronic Medical Systems, Bergdorf, Switzerland), HUMALOG MIX <NUM>/<NUM>™ pen, HUMALOG™ pen, HUMALIN <NUM>/<NUM>™ pen (Eli Lilly and Co. , Indianapolis, IN), NOVOPEN™ I, II and III (Novo Nordisk, Copenhagen, Denmark), NOVOPEN JUNIOR™ (Novo Nordisk, Copenhagen, Denmark), BD™ pen (Becton Dickinson, Franklin Lakes, NJ), OPTIPEN™, OPTIPEN PRO™, OPTIPEN STARLET™, and OPTICLIK™ (sanofi-aventis, Frankfurt, Germany), to name only a few. Examples of disposable pen and/or autoinjector delivery devices having applications in subcutaneous delivery of a pharmaceutical composition described herein include, but are not limited to the SOLOSTAR™ pen (sanofi-aventis), the FLEXPEN™ (Novo Nordisk), and the KWIKPEN™ (Eli Lilly), the SURECLICK™ Autoinjector (Amgen, Thousand Oaks, CA), the PENLET™ (Haselmeier, Stuttgart, Germany), the EPIPEN (Dey, L. ), and the HUMIRA™ Pen (Abbott Labs, Abbott Park, IL), to name only a few.

The use of a microinfusor to deliver the pharmaceutical formulations described herein is also contemplated herein. As used herein, the term "microinfusor" means a subcutaneous delivery device designed to slowly administer large volumes (e.g., up to about <NUM> or more) of a therapeutic formulation over a prolonged period of time (e.g., about <NUM>, <NUM>, <NUM>, <NUM>, <NUM> or more minutes). See, e.g., <CIT>; <CIT>; and <NPL>). Microinfusors are particularly useful for the delivery of large doses of therapeutic proteins contained within high concentration (e.g., about <NUM>, <NUM>, <NUM>, <NUM>, <NUM> or more mg/mL) and/or viscous solutions.

The pharmaceutical formulations of the present invention are useful, inter alia, for the treatment, prevention and/or amelioration of any disease or disorder associated with IL-<NUM> activity, including diseases or disorders mediated by activation of the IL-<NUM> receptor. Exemplary, non-limiting diseases and disorders that can be treated and/or prevented by the administration of the pharmaceutical formulations of the present invention include, e.g., rheumatoid arthritis, ankylosing spondylitis, Crohn's disease, ulcerative colitis, pancreatitis, juvenile idiopathic arthritis, vasculitis, Kawasaki disease, systemic lupus erythematosis, psoriasis, psoriatic arthritis, Sjogren syndrome, Still's disease, Castleman's disease, multiple sclerosis, diseases associated with abnormal blood coagulation or fibrinolysis (e.g., thrombosis), cancer (e.g., breast cancer, leukemia, ovarian cancer, melanoma, prostate cancer, pancreatic cancer, lymphoma, lung cancer, renal cell carcinoma, colorectal cancer, multiple myeloma, etc.), cachexia, chronic rejection of transplanted organs and cells, cardiopathy, viral infection (e.g., HIV infection, EBV infection, etc.), plasmacytosis, hyperimmunoglobulinemia, anemia, nephritis, mesothelioma, and hearing loss and other inner ear disorders.

Thus, the present invention provides use of the formulation defined in the claims in a method of treating, preventing, and/or ameliorating any disease or disorder associated with IL-<NUM> activity or IL-6R activation (including any of the above mentioned exemplary diseases, disorders and conditions). The subject to which the pharmaceutical formulation is administered can be, e.g., any human or non-human animal that is in need of such treatment, prevention and/or amelioration, or who would otherwise benefit from the inhibition or attenuation of IL-<NUM> and/or IL-6R-mediated activity. For example, the subject can be an individual that is diagnosed with, or who is deemed to be at risk of being afflicted by any of the aforementioned diseases or disorders. The present disclosure further includes the use of any of the pharmaceutical formulations disclosed herein in the manufacture of a medicament for the treatment, prevention and/or amelioration of any disease or disorder associated with IL-<NUM> activity or IL-6R activation (including any of the above mentioned exemplary diseases, disorders and conditions).

The following examples are put forth so as to provide those of ordinary skill in the art with a complete disclosure and description of how to make and use the methods and compositions of the invention, and are not intended to limit the scope of what the inventors regard as their invention. Efforts have been made to ensure accuracy with respect to numbers used (e.g., amounts, temperature, etc.) but some experimental errors and deviations should be accounted for. Unless indicated otherwise, parts are parts by weight, molecular weight is average molecular weight, temperature is in degrees Centigrade, and pressure is at or near atmospheric.

In this Example, various formulations were created containing an anti-human IL-6R antibody without excipients. The exemplary antibody used in this and all subsequent Examples set forth below is an antibody comprising a heavy chain variable region (HCVR) with the amino acid sequence of SEQ ID NO:<NUM>, and a light chain variable region (LCVR) with the amino acid sequence of SEQ ID NO:<NUM>. This antibody is referred to herein as "mAb1".

As a preliminary experiment, the stability of mAb1 in liquid solution was determined following various amounts of time in frozen storage at -<NUM>, -<NUM> and -<NUM>. The concentration of mAb1 used in this Example was <NUM>/mL. At various time points, the stability of mAb1 was determined by size exclusion high performance liquid chromatography (SE-HPLC) and by cation exchange high performance liquid chromatography (CEX-HPLC). Stability was assessed based on the percentage of native mAb1 remaining in the sample (by SE-HPLC; Table <NUM>) and by the percentage of acidic species observed in the sample (by CEX-HPLC; Table <NUM>) (An increase in percent acidic species is consistent with deamidation of the antibody and is thus considered an undesired phenomenon with respect to the pharmaceutical formulations of the present disclosure).

The results of Tables <NUM> and <NUM> are depicted in <FIG> and <FIG>, respectively. These results show that mAb1 can remain stable at a concentration of <NUM>/mL for at least <NUM> months when stored at -<NUM>.

Eight different formulations containing mAb1 and minimal excipients as shown in Table <NUM> were prepared.

The formulations were tested for stability by SE-HPLC after various amounts of time at - <NUM> and -<NUM>. The results, expressed in percent of native mAb1 remaining, are shown in Tables <NUM> (-<NUM> storage) and <NUM> (-<NUM>).

The results of Tables <NUM> and <NUM> are depicted in <FIG> and <FIG>, respectively. As shown in this Example, the stability of mAb1 was maintained to a significant extent in formulations <NUM>, <NUM>, <NUM> and <NUM> after several months of storage at -<NUM> and -<NUM>. These results indicate that the stability of mAb1 at -<NUM> and -<NUM> can be enhanced by the addition of at least <NUM>% sucrose.

A stabilized formulation containing various concentrations of mAb1 was prepared for use in Examples <NUM> and <NUM> below. This formulation, designated "Formulation A", is shown in Table <NUM>.

Formulation A (see Example <NUM>) containing <NUM>, <NUM> or <NUM>/mL mAb1 was tested for stability after several months of storage at <NUM> in clear vials. Stability was assessed by the following parameters: (a) visual appearance; (b) turbidity (OD <NUM>); (c) pH; (d) percent total mAb1 recovered (as measured by RP-HPLC); (d) percent native mAb1 recovered (as measured by SE-HPLC); (e) percent main peak mAb1 recovered (as measured by CEX-HPLC); and (f) percent acidic species mAb1 recovered (as measured by CEX-HPLC). The stability results for Formulation A containing <NUM>, <NUM> and <NUM>/mL of mAb1 are summarized in Tables <NUM>, <NUM> and <NUM>, respectively.

The results of this Example demonstrate that Formulation A containing <NUM>, <NUM> or <NUM>/mL mAb1 remained stable after at least <NUM> months of storage, at <NUM> in clear vials, with about <NUM>% or more of native mAb1 remaining in all samples after <NUM> months of storage under such conditions. For the <NUM> and <NUM>/mL formulations, <NUM>% and <NUM>% of native mAb1, respectively, was detected after up to <NUM> months of storage at <NUM>. In addition, the percent acidic species remained at <NUM>% or lower for all time points analyzed, thus confirming the stability of the formulations.

Similar stability studies were also carried out using Formulation A containing <NUM>/mL mAb1 following storage at <NUM>-<NUM>. No significant degradation was observed for any of the concentrations tested after <NUM> months of <NUM>-<NUM> storage as determined by SE-HPLC and CEX-HPLC (data not shown).

Additional experiments were conducted to compare the stability of Formulation A (see Example <NUM>) containing <NUM> and <NUM>/mL mAb1 manufactured in amber glass vials to the same formulation manufactured in clear vials. Two types of amber vials were used in this Example: <NUM> and <NUM> amber vials. Stability was assessed following storage at <NUM>, <NUM> or <NUM> based on the same parameters as used in Example <NUM>. The results for the <NUM>/mL and <NUM>/mL formulations are summarized in Tables <NUM> through <NUM>.

As shown in this Example, Formulation A containing <NUM>/mL or <NUM>/mL mAb1 exhibited equivalent stability profiles when stored in either clear or amber vials. Moreover, as demonstrated in Example <NUM> for storage in clear vials, relatively high stability of mAb1 was maintained in Formulation A when stored in either clear or amber vials at <NUM> for up to <NUM> months.

Several formulations were prepared containing <NUM>/mL, <NUM>/mL and <NUM>/mL mAb1 and various quantities of histidine, arginine and sucrose. Viscosity and osmolality were measured for each formulation. Additionally, the stability of the <NUM>/mL formulations after <NUM> weeks of storage at <NUM> was assessed in terms of percent native mAb1 remaining (by SE-HPLC) and percent main peak remaining (by CEX-HPLC). The results are summarized in Table <NUM>. The formulations presented in Table <NUM> are not in the scope of the claims.

The results presented in Table <NUM> indicate that increasing the histidine concentration to <NUM> or <NUM> and adding arginine to the formulation (<NUM> or <NUM>) significantly reduced the viscosity of the formulation as compared to formulations containing only <NUM> histidine and no arginine. Furthermore, reducing the sucrose concentration from <NUM>% to <NUM>% with the added histidine and arginine decreased the viscosity of the formulation to an even greater extent.

Based at least in part on the foregoing, the following Formulations (designated "Formulation B" and "Formulation C") set forth in Table <NUM> were prepared.

Formulation B (see Table <NUM>) containing <NUM>/mL mAb1 was prepared in a <NUM> glass vial and in two different syringes: regular and low tungsten. The preparations were stored at <NUM>, <NUM> and <NUM> for various amounts of time. The stability of mAb1 following storage was measured by SE-HPLC and CEX-HPLC. The results are shown in Table <NUM>. (An increase in percent acidic species is consistent with deamidation of the antibody and is thus considered an undesired phenomenon with respect to the pharmaceutical formulations of the present disclosure).

As shown in this table, Formulation B containing <NUM>/mL mAb1, stored at <NUM> in a glass vial or syringe, remained relatively stable for at least <NUM> months.

A series of experiments was carried out to assess the stability of different mAb1 formulations in prefilled syringes. For these experiments various luer and staked needle, regular-tungsten and low-tungsten syringes were used in combination with different types of plungers (coated and uncoated) and tip-caps. The formulations were tested for stability after storage in prefilled syringes at <NUM>, <NUM> and <NUM> for various amounts of time (ranging from <NUM> days to <NUM> months, depending on the conditions tested).

Six different formulations of mAb1 were tested for stability in prefilled syringes in this Example: (<NUM>) Formulation A (see Table <NUM>) containing <NUM>/mL mAb1; (<NUM>) Formulation A (see Table <NUM>) containing <NUM>/mL mAb1; (<NUM>) Formulation B (see Table <NUM>) containing <NUM>/mL mAb1; (<NUM>) Formulation B (see Table <NUM>) containing <NUM>/mL mAb1; (<NUM>) Formulation C (see Table <NUM>) containing <NUM>/mL of mAb1; and (<NUM>) Formulation C (see Table <NUM>) containing <NUM>/mL of mAb1.

Stability was assessed by the following parameters: (a) visual analysis; (b) turbidity (OD<NUM>); (c) percent recovery by RP-HPLC; (d) percent native mAb1 by SE-HPLC; (e) percent main peak mAb1 by CEX-HPLC; and (f) percent acidic species by CEX-HPLC.

The results from a representative experiment assessing the stability of Formulation A, containing <NUM>/mL mAb1 in two different syringes (Syringe #<NUM> and Syringe #<NUM>) are shown in Tables <NUM> and <NUM> below.

The results from another representative experiment assessing the stability of Formulation C, containing <NUM>/mL mAb1 in two different syringes (Syringe #<NUM> and Syringe #<NUM>) are shown in Tables <NUM> and <NUM> below.

The results from this set of experiments demonstrate that the different formulations remain relatively stable in prefilled syringes, especially when stored at temperatures of <NUM> and below, for one month or greater. Moreover, the various formulations disclosed herein appeared to have enhanced stability when contained in low tungsten syringes containing fluorocarbon-coated plungers.

Claim 1:
A pharmaceutical formulation for use in a method of treatment, prevention and/or amelioration of any disease or disorder associated with IL-<NUM> activity, wherein the method comprises injection of the pharmaceutical formulation into the subject, and wherein the pharmaceutical formulation comprises:
(i) a human antibody that specifically binds to human interleukin-<NUM> receptor (hIL-6R), wherein the antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: <NUM> and a light chain variable region comprising the amino acid sequence of SEQ ID NO:<NUM>;
(ii) histidine at a concentration of from <NUM> to <NUM>;
(iii) arginine at a concentration of from <NUM> to <NUM>;
(iv) sucrose in an amount of from <NUM>% to <NUM>% w/v; and
(v) polysorbate <NUM> in an amount of from <NUM>% to <NUM>% w/v.