Anti-CD20 antibody formulations

Abstract

The present application relates to aqueous pharmaceutical formulations for subcutaneous administration comprising an anti-CD20 antibody at a concentration of 100 mg / ml to 120 mg / ml. The formulation further comprises sodium acetate, sucrose, methionine, polysorbate, hyaluronidase, and polyvinylpyrrolidone (PVP) K12. Articles of manufacture that comprise such formulations and methods of treatment that involve the use of such formulations are also included.

Description

Attorney Docket No. 146392068840ANTI-CD20 ANTIBODY FORMULATIONSCROSS REFERENCE TO RLATED APPLICATIONS

[0001] This application claims priority to and the benefit of U.S. Provisional Application No. 63 / 714,501, filed on October 31, 2024, the contents of which are incorporated herein by reference in its entirety.REFERENCE TO AN ELECTRONIC SEQUENCE LISTING

[0002] The contents of the electronic sequence listing (146392068840SEQLIST.xml; Size: 11,860 bytes; and Date of Creation: October 22, 2025) is herein incorporated by reference in its entirety.FIELD OF THE INVENTION

[0003] The present disclosure relates to formulations for subcutaneous administration of an anti-CD20 antibody, articles of manufacture comprising such formulations, and methods for treating multiple sclerosis (MS) in a patient with such formulations.BACKGROUND OF THE INVENTION

[0004] To improve patient convenience and compliance with monoclonal antibody (mAb) based pharmaceutical treatments, there is a desire for means of administration in addition to intravenous (i.v. or IV) administration by healthcare professionals. One such means is subcutaneous (s.c. or SC) administration, which in certain cases may be self-administered. Smaller volumes are preferred for SC administration which makes highly concentrated antibody formulation more favorable. However, multiple challenges are presented during the development of a highly concentrated antibody formulation that can be used for SC administration into a subject. Depending on the specific physiochemical and biological properties of a particular antibody molecule, such challenges include solubility in both the stock solution and the subcutaneous space of the subject, stability, toxicity, bioavailability, and immunogenicity. See e.g., J Pharm Sci. 2022 Apr;l l l(4):861-867.

[0005] Ocrelizumab (“OCR”) is a recombinant, humanized, monoclonal IgGl antibody that selectively targets and depletes CD20-expressing B cells, while preserving the capacity of B- cell reconstitution and preexisting humoral immunity. CD20 is a B-cell surface molecule that is restricted in expression to pre-B cells and mature B cells but is not expressed earlier in the development of B cells. Ocrelizumab administered by intravenous (IV) infusion at a dose of1MF-363724269Attorney Docket No. 146392068840600 mg every 6 months was approved by the U.S. Food and Drug Administration (FDA) in 2017 for the treatment of adult patients with multiple sclerosis.

[0006] All references cited herein, including patent applications, patent publications, and UniProtKB / Swiss-Prot Accession numbers are herein incorporated by reference in their entirety, as if each individual reference were specifically and individually indicated to be incorporated by reference.SUMMARY OF THE APPLICATION

[0007] The present application in one aspect provides an aqueous pharmaceutical formulation, the formulation comprising an anti-CD20 antibody in a concentration of about 99 mg / ml to about 121 mg / ml, hyaluronidase in a concentration of at least about 500 U / ml, polyvinylpyrrolidone (PVP) K12 in a concentration of about 5% (w / v) to about 10% (w / v), methionine in a concentration of about 10 mM to about 50 mM, sucrose or trehalose, a surfactant, and a buffer, wherein the formulation has a pH of about 5.0 to about 5.6, wherein the anti-CD20 antibody comprises a VH domain comprising the amino acid sequence set forth in SEQ ID NO: 8, a VL domain comprising the amino acid sequence set forth in SEQ ID NO: 7, and a human IgGl constant region, wherein the formulation is suitable for subcutaneous administration. In some embodiments, hyaluronidase is in a concentration of about 1000 U / ml to about 3000 U / ml. In some embodiments, hyaluronidase is in a concentration of about 1500 U / ml to about 2500 U / ml. In some embodiments, hyaluronidase is in a concentration of about 1800 U / ml to about 2200 U / ml. In some embodiments, hyaluronidase is in a concentration of about 2000 U / ml.

[0008] In some embodiments according to any of the formulations described above, the anti- CD20 antibody is in a concentration of about 110 mg / ml.

[0009] In some embodiments according to any of the formulations described above, the PVP K12 is in a concentration of about 5% (w / v) to about 7.5% (w / v). In some embodiments, the PVP K12 is in a concentration of about 7.5% (w / v).

[0010] In some embodiments according to any of the formulations described above, the hyaluronidase is in a concentration of about 2000 U / ml.

[0011] In some embodiments according to any of the formulations described above, methionine is in a concentration of about 10 mM to about 40 mM. In some embodiments,2MF-363724269Attorney Docket No. 146392068840 methionine is in a concentration of about 25 mM to about 35 mM. In some embodiments, methionine is in a concentration of about 30 mM.

[0012] In some embodiments according to any of the formulations described above, the formulation comprises sucrose. In some embodiments, the sucrose is in a concentration of about 150 mM to about 210 mM. In some embodiments, the sucrose is in a concentration of about 180 mM.

[0013] In some embodiments according to any of the formulations described above, the surfactant is polysorbate 20. In some embodiments, the polysorbate is in a concentration of at least 0.03% (w / v). In some embodiments, the polysorbate is in a concentration of about 0.03% (w / v) to about 0.08% (w / v) or about 0.04% (w / v) to about 0.08% (w / v). In some embodiments, the polysorbate is in a concentration of about 0.06% (w / v).

[0014] In some embodiments according to any of the formulations described above, the pH is about 5.3.

[0015] In some embodiments according to any of the formulations described above, the buffer in the formulation is an acetate buffer. In some embodiments, the acetate buffer has an acetate concentration of about 20 mM to about 50 mM. In some embodiments, the acetate buffer has an acetate concentration of about 37 mM to about 47 mM. In some embodiments, the acetate buffer has an acetate concentration of about 43 mM.

[0016] In some embodiments according to any of the formulations described above, the buffer is a sodium acetate buffer. In some embodiments, the sodium acetate buffer has a sodium concentration of about 2 mM to about 8 mM and an acetate concentration of about 37 mM to about 47 mM. In some embodiments, the sodium acetate buffer has a sodium concentration of about 5 mM and an acetate concentration of about 43 mM.

[0017] The present application in another aspect provides an aqueous pharmaceutical formulation, the formulation comprising an anti-CD20 antibody in a concentration of 110 mg / ml, sucrose in a concentration of 180 mM, methionine in a concentration of 30 mM, polysorbate 20 in a concentration of 0.06% (w / v), hyaluronidase in a concentration of 2000 U / ml, polyvinylpyrrolidone (PVP) K12 in a concentration of about 7.5% (w / v), and an acetate buffer, and wherein the formulation has a pH of 5.0 to 5.6 (e.g., about 5.3), wherein the anti-CD20 antibody comprises a VH domain comprising the amino acid sequence set forth in SEQ ID NO: 8, a VL domain comprising the amino acid sequence set forth in SEQ3MF-363724269Attorney Docket No. 146392068840ID NO: 7, and a human IgGl constant region, wherein the formulation is suitable for subcutaneous administration. In some embodiments, the formulation has a pH of 5.3.

[0018] In some embodiments according to any of the formulations described above, the formulation has a sodium acetate buffer, and wherein the sodium acetate buffer has a sodium concentration of 5 mM and an acetate concentration of 43 mM.

[0019] In some embodiments according to any of the formulations described above, the weight average molecular weight of PVP K12 is 2000 Dalton to 7000 Dalton. In some embodiments, the molecular weight is assessed by size exclusion chromatography (SEC) using light scattering detection. In some embodiments, the weight average molecular weight of PVP K12 is 2000 Dalton to 3000 Dalton.

[0020] In some embodiments according to any of the formulations described above, the anti- CD20 antibody comprises a light chain comprising the amino acid sequence of SEQ ID NO: 9 and a heavy chain comprising the amino acid sequence of SEQ ID NO: 10 or 11.

[0021] In some embodiments according to any of the formulations described above, the anti- CD20 antibody is ocrelizumab.

[0022] In some embodiments according to any of the formulations described above, the hyaluronidase is recombinant human hyaluronidase.

[0023] In some embodiments according to any of the formulations described above, a) the antibody in the formulation is stable at 2-8°C for at least 6 months, at least 12 months, at least 18 months or at least 24 months, b) the antibody and hyaluronidase in the formulation are stable after continuous agitation for 24 hours at room temperature, and / or c) there is i) less than 1% of aggregation of the antibody in the formulation as assessed by sum of HMW forms by SE-UHPLC after 25 days at 40°C and / or ii) less than 6% of degradation of the antibody in the formulation as measured by sum of pre-peaks by CE-SDS after 25 days at 40°C.

[0024] In some embodiments according to any of the formulations described above, the hyaluronidase in the formulation is stable when it retains at least about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of its biological activity.

[0025] In some embodiments according to any of the formulations described above, the antibody in the formulation is stable when it retains at least about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of its biological activity.4MF-363724269Attorney Docket No. 146392068840

[0026] In some embodiments according to any of the formulations described above, the biological activity is measured by antibody binding to CD20.

[0027] In some embodiments according to any of the formulations described above, the formulation is sterile.

[0028] The present application in another aspect provides a method of promoting solubility and bioavailability of an anti-CD20 antibody in an aqueous formulation, comprising adding a buffer comprising polyvinylpyrrolidone (PVP) K12 into a solution comprising the anti-CD20 antibody for preparation of the formulation, wherein the formulation comprises the antibody in a concentration of about 100 mg / ml to about 130 mg / ml and the PVP K12 in a concentration of about 7.5% (w / v).

[0029] In some embodiments according to any of the methods described above, the formulation further comprises hyaluronidase.

[0030] The present application in another aspect provides a method of protecting an aqueous pharmaceutical formulation comprising an anti-CD20 antibody and hyaluronidase from agitation stress, comprising adding a buffer comprising polyvinylpyrrolidone (PVP) K12 into a solution comprising the anti-CD20 antibody for preparation of the formulation, wherein the formulation comprises the PVP K12 in a concentration of at least about 5% (w / v).

[0031] In some embodiments according to any of the methods described above, the PVP KI 2 has a concentration of between about 5% (w / v) and about 10% (w / v). In some embodiments, the PVP K12 has a concentration of about 7.5% (w / v).

[0032] In some embodiments according to any of the methods described above, hyaluronidase is in a concentration of about 1000 U / ml to about 3000 U / ml. In some embodiments, hyaluronidase is in a concentration of about 1500 U / ml to about 2500 U / ml. In some embodiments, the hyaluronidase is in a concentration of about 1800 U / ml to about 2200 U / ml. In some embodiments, the hyaluronidase is in a concentration of about 2000 U / ml. In some embodiments, the hyaluronidase is a recombinant human hyaluronidase.

[0033] In some embodiments according to any of the methods described above, the anti- CD20 antibody is in a concentration of about 110 mg / ml.

[0034] In some embodiments according to any of the methods described above, the formulation further comprises methionine. In some embodiments, methionine is in a concentration of about 10 mM to about 50 mM, or about 10 mM to about 40 mM, or about 255MF-363724269Attorney Docket No. 146392068840 mM to about 35 mM. In some embodiments, methionine is in a concentration of about 30 mM.

[0035] In some embodiments according to any of the methods described above, the formulation comprises sucrose or trehalose.

[0036] In some embodiments according to any of the methods described above, the formulation comprises sucrose. In some embodiments, the formulation comprises sucrose in a concentration of about 150 mM to about 210 mM. In some embodiments, the sucrose is in a concentration of about 180 mM.

[0037] In some embodiments according to any of the methods described above, the surfactant is polysorbate. In some embodiments, the surfactant is polysorbate 20.

[0038] In some embodiments according to any of the methods described above, the polysorbate is in a concentration of about 0.04% (w / v) to about 0.08% (w / v). In some embodiments, the polysorbate is in a concentration of about 0.06% (w / v).

[0039] In some embodiments according to any of the methods described above, the pH is about 4.8 to about 5.8. In some embodiments, the pH is about 5.0 to about 5.6. In some embodiments, the pH is about 5.3.

[0040] In some embodiments according to any of the methods described above, the formulation comprises a sodium acetate buffer. In some embodiments, the sodium acetate buffer has an acetate concentration of about 20 mM to about 50 mM. In some embodiments the acetate concentration is about 37 mM to about 47 mM. In some embodiments, the acetate concentration is 43 mM. In some embodiments the sodium acetate buffer has a sodium concentration of about 2 mM to about 8 mM. In some embodiments, the sodium concentration is 5 mM.

[0041] In some embodiments according to any of the methods described above, the formulation comprises an anti-CD20 antibody in a concentration of about 110 mg / ml, sucrose in a concentration of about 180 mM, methionine in a concentration of about 30 mM, polysorbate 20 in a concentration of about 0.06% (w / v), hyaluronidase in a concentration of about 2000 U / ml, and PVP K12 in a concentration of about 7.5% (w / v), a sodium acetate buffer having a sodium concentration of about 5 mM and an acetate concentration of about 43 mM, and wherein the formulation has a pH of about 5.3.6MF-363724269Attorney Docket No. 146392068840

[0042] In some embodiments according to any of the methods described above, the weight average molecular weight of PVP K12 is 2000 Dalton to 7000 Dalton. In some embodiments, the molecular weight is assessed by size exclusion chromatography (SEC) using light scattering detection. In some embodiments, the weight average molecular weight of PVP K12 is 2000 Dalton to 3000 Dalton.

[0043] In some embodiments according to any of the methods described above, the anti- CD20 antibody comprises a VH domain comprising the amino acid sequence set forth in SEQ ID NO: 8, a VL domain comprising the amino acid sequence set forth in SEQ ID NO: 7, and a human IgGl constant region, and wherein the formulation is suitable for subcutaneous administration.

[0044] In some embodiments according to any of the methods described above, the anti- CD20 antibody comprises a light chain comprising the amino acid sequence of SEQ ID NO: 9 and a heavy chain comprising the amino acid sequence of SEQ ID NO: 10 or 11.

[0045] The present application in another aspect provides a method of treating multiple sclerosis in a patient comprising subcutaneously administering the formulation of any one of any of the formulations described above.

[0046] In some embodiments according to any of the methods described above, the patient has not been subjected to a prior intravenous administration of the anti-CD20 antibody at a dose of about 600 mg.

[0047] In some embodiments according to any of the methods described above, the patient has not been subjected to a prior intravenous administration of the anti-CD20 antibody.

[0048] In some embodiments according to any of the methods described above, the patient has not been subjected to a prior subcutaneous administration of the anti-CD20 antibody at a dose lower than about 900 mg.

[0049] In some embodiments according to any of the methods described above, the patient has received a prior treatment with an anti-CD20 antibody. In some embodiments, the prior treatment comprises an intravenous administration of the anti-CD20 antibody. In some embodiments, the prior treatment comprises the intravenous administration of the anti-CD20 antibody at a dose of about 300 mg or about 600 mg.

[0050] In some embodiments according to any of the methods described above, the method comprises subcutaneously administering more than one dose of the anti-CD20 antibody.7MF-363724269Attorney Docket No. 146392068840

[0051] In some embodiments according to any of the methods described above, the method comprises administering at least two, three or four doses of the anti-CD20 antibody.

[0052] In some embodiments according to any of the methods described above, the anti- CD20 antibody is administered at a frequency of no more than once every 24 weeks or about once every 24 weeks.

[0053] In some embodiments according to any of the methods described above, the anti- CD20 antibody is administered at a frequency of no more than once every 6 months or about once every 6 months.

[0054] In some embodiments according to any of the methods described above, anti-CD20 antibody is the only medicament administered to the patient to treat multiple sclerosis.

[0055] In some embodiments according to any of the methods described above, the patient is subject to a) an oral dexamethasone or an equivalent corticosteroid and b) an oral antihistamine administration (e.g., desloratadine) shortly prior to the subcutaneous administration of the anti-CD20 antibody. In some embodiments, the oral dexamethasone or the equivalent corticosteroid and antihistamine are administered at least or within 30 minutes, 20 minutes, or 15 minutes prior to the subcutaneous administration. In some embodiments, the patient is subject to 20 mg dexamethasone and 5 mg desloratadine.

[0056] In some embodiments according to any of the methods described above, the multiple sclerosis is relapsing multiple sclerosis (RMS).

[0057] In some embodiments according to any of the methods described above, the patient has a clinical isolated syndrome (CIS), relapsing-remitting multiple sclerosis (RRMS) or active secondary progressive multiple sclerosis (SPMS).

[0058] In some embodiments according to any of the methods described above, the multiple sclerosis is a primary progressive multiple sclerosis (PPMS).

[0059] In some embodiments according to any of the methods described above, the patient has an Expanded Disability Status Scale (EDSS) score of 0-6.5.

[0060] In some embodiments according to any of the methods described above, the subcutaneous administration takes no more than about 10 minutes.

[0061] In some embodiments according to any of the methods described above, said anti- CD20 antibody in the formulation is about 110 mg / ml.8MF-363724269Attorney Docket No. 146392068840

[0062] In some embodiments according to any of the methods described above, the anti- CD20 antibody comprises a light chain comprising the amino acid sequence of SEQ ID NO: 9 and a heavy chain comprising the amino acid sequence of SEQ ID NO: 10 or 11.

[0063] In some embodiments according to any of the methods described above, the anti- CD20 antibody is ocrelizumab.

[0064] It is to be understood that one, some, or all of the properties of the various embodiments described herein may be combined to form other embodiments of the present application. These and other aspects of the application will become apparent to one of skill in the art. These and other embodiments of the application are further described by the detailed description that follows.BRIEF DESCRIPTION OF THE DRAWINGS

[0065] The drawings illustrate certain features and advantages of this disclosure. These embodiments are not intended to limit the scope of the appended claims in any manner.

[0066] FIG. 1 shows the study design of the Study B described in Example 2.

[0067] FIG. 2 shows incidence and severity of microscopic findings at subcutaneous site B on Study Day 1, Day 15, and Day 29 in Study B.

[0068] FIG. 3 shows the concentration of ocrelizumab in male rat serum on Study Day 1 or Study Day 15 in Study B.

[0069] FIG. 4 shows Cmax, AUC and T1 / 2 after first and second administration in Study B.

[0070] FIG. 5 shows size exclusion HPLC results of ocrelizumab. Absorbance at 280 (Left) and radioactivity (Right) from SEC-HPLC are shown. Retention times for single peaks are listed in the upper right corner of the control traces, test radiolabels and for dose material as indicated on the left text panels. Y-axis is percent relative response and X-axis is retention time (min).

[0071] FIG. 6 shows SPECT-CT imaging of ocrelizumab in rats subcutaneously injected with ocrelizumab. SPECT-CT images of each group are shown. Antibody concentrations were depicted using a concentration-based color map with black representing 0% ID / g and red representing the highest signal of 100 %ID / g at 2 h, 50% ID / g at 1 d, 25% ID / g at 2 d and 15% ID / g at 3 d. All 3 rats per group are shown at each time point. Dose group information is9MF-363724269Attorney Docket No. 146392068840 on the far-left panel. All animals are positioned on their sides to have the interscapular subcutaneous injection site on the left side of the image.

[0072] FIG. 7 shows SPECT SC bolus quantification in rats subcutaneously injected with ocrelizumab. SC bolus volumes of interest (VOI) were segmented from a larger VOI of the back of the animal ranging from the base of the neck to the bottom of the rib cage from images shown in FIG. 6. The SC bolus was then segmented by thresholding the signal activity to be greater than or equal to 1 %ID / g. The data was then converted to %ID based on the volume of the VOI, assuming the tissue density to be that of water. Error bars are representing standard deviation, and each sample is the average of the group (n=3). Time points analyzed are listed in Table 9.

[0073] FIG. 8 shows plasma exposure of ocrelizumab in rats subcutaneously injected with ocrelizumab. Plasma concentration time profiles. Time points analyzed are listed in Table 9. Error bars are representing standard deviation, and each sample is the average of the group (n=3). The y-axis is plotted on log scale.

[0074] FIG. 9 shows PVP K12 enhances solubility of ocrelizumab high concentration formulations in a turbidity plate screening assay.

[0075] FIG. 10A shows vial agitation study results.

[0076] FIG. 10B shows loss of rHuPH20 activity under agitation stress.

[0077] FIG. 11 shows an overview of the surface tension measurements using different conditions of ocrelizumab formulation buffers, and the drug products.

[0078] FIG. 12 shows analytical data for ocrelizumab SC 40 mg / mL Ocrevus Zunovo® (triangle) and ocrelizumab high concentration (or “HC”) (circle).

[0079] FIGs. 13A-13B show stress comparability between ocrelizumab HC formulated with 240 mM sucrose (Phi DP) and 180 mM sucrose (Phlll DP) by IE-HPLC. Main peak: FIG. 13A; Acidic peak: FIG. 13B.

[0080] FIGs. 14A-14B show stress comparability between ocrelizumab HC formulated with 240 mM sucrose (Phi DP) and 180 mM sucrose (Phlll DP) by SE-UHPLC. Main peak: FIG. 14A; Sum of HMW forms: FIG. 14B.

[0081] FIGs. 15A-15B show stress comparability between ocrelizumab HC formulated with 240 mM sucrose (Phi DP) and 180 mM sucrose (Phlll DP) by non-reduced CE-SDS. Main peak: FIG. 15A; Sum of pre-peaks (LMW forms): FIG. 15B.10MF-363724269Attorney Docket No. 146392068840

[0082] FIGs. 16A-16C show stability from three lots of ocrelizumab high concentration formulations after 5x F / T at -20°C with the main peak by SE-UHPLC (FIG. 16A), sum of HMW forms by SE-UHPLC (FIG. 16B), and overlaid chromatograms from SE-UHPLC analysis (FIG. 16C).

[0083] FIGs. 17A-17C show stability from three lots of ocrelizumab high concentration formulations after frozen storage at -8°C for 30 days with the main peak by SE-UHPLC (FIG. 17A), sum of HMW forms by SE-UHPLC (FIG. 17B), and overlaid chromatograms from SE-UHPLC analysis (FIG. 17C).

[0084] FIG. 18 shows PS20 concentrations, instances of visible particles, and subvisible particle count for both PCL and CALB enzymes.

[0085] FIGs. 19A-19C show pictures of dialysis cassettes from the buffer exchange at TO (FIG. 19A), 48 hours (FIG. 19B), and 72 hours (FIG. 19C) at room temperature.DETAILED DESCRIPTION

[0086] The present application provides an aqueous pharmaceutical formulation comprising a high concentration of an anti-CD20 antibody ocrelizumab for subcutaneous administration. The pharmaceutical formulation further comprises hyaluronidase, polyvinylpyrrolidone (PVP), methionine, a stabilizer, a surfactant and a buffer that provides a pH of about 5.0 to about 5.6.

[0087] The present application in one aspect provides an aqueous pharmaceutical formulation, the formulation comprising an anti-CD20 antibody in a concentration of about 99 mg / ml to 121 mg / ml, hyaluronidase in a concentration of at least about 500, polyvinylpyrrolidone (PVP) K12 in a concentration of about 5% (w / v) to about 10% (w / v), methionine in a concentration of about 10 mM to about 50 mM, sucrose or trehalose, a surfactant, and a buffer, wherein the formulation has a pH of about 5.0 to about 5.6, wherein the anti-CD20 antibody comprises a VH domain comprising the amino acid sequence set forth in SEQ ID NO: 8, a VL domain comprising the amino acid sequence set forth in SEQ ID NO: 7, and a human IgGl constant region, wherein the formulation is suitable for subcutaneous administration. In some embodiments, hyaluronidase is in a concentration of about 1000 U / ml to about 3000 U / ml. In some embodiments, hyaluronidase is in a concentration of about 1500 U / ml to about 2500 U / ml. In some embodiments, hyaluronidase11MF-363724269Attorney Docket No. 146392068840 is in a concentration of about 1800 U / ml to about 2200 U / ml. In some embodiments, hyaluronidase is in a concentration of about 2000 U / ml.

[0088] The present application further provides an aqueous pharmaceutical formulation comprising an anti-CD20 antibody in a concentration of about 110 mg / ml to about 120 mg / ml, sodium acetate in a concentration of about 20 mM, sucrose in a concentration of about 180 mM to about 240 mM, methionine in a concentration of about 30 mM, polysorbate (e.g., polysorbate 20) in a concentration of about 0.06% (w / v), hyaluronidase in a concentration of about 2000 U / ml, and polyvinylpyrrolidone (PVP) K12 in a concentration of about 7.5% (w / v), and wherein the formulation has a pH of about 5.3, wherein the anti-CD20 antibody comprises a VH domain comprising the amino acid sequence set forth in SEQ ID NO: 8, a VL domain comprising the amino acid sequence set forth in SEQ ID NO: 7, and a human IgGl constant region, and wherein the formulation is suitable for subcutaneous administration. In some embodiments, the anti-CD20 antibody is ocrelizumab.

[0089] The present application is at least partly based upon inventors’ findings that the high concentration ocrelizumab (e.g., 110-120 mg / mL) formulations described herein achieved multiple advantageous effects for subcutaneous administration in subjects such as human patients. These advantageous effects include a) high concentration ocrelizumab that is soluble not only in storage solution but also in subcutaneous space after it is administered, b) minimum toxicity and c) desirable bioavailability. See e.g. Examples 2, 3, 4, and 6. A representative formulation is as described in Table 1 of Example 1. Furthermore, the present application provides PVP KI 2 surprisingly a) acts as an effective surfactant and protects both ocrelizumab and hyaluronidase from agitate stress (see e.g., Example 5), b) acts as an effective cryoprotectant and protects ocrelizumab from multiple freeze and thaw (see e.g., Example 8), c) protects degradation of polysorbate degradation (see e.g., Example 9). It was also surprisingly found that an anti-CD20 formulation with sucrose at a concentration of 180 mM is more effective in preventing aggregation and degradation as compared to a similar formulation with sucrose at a concentration of 240 mM. See e.g., Example 7A.

[0090] The present application further provides articles of manufacture comprising such formulations and methods of treatment comprising administering such formulations.12MF-363724269Attorney Docket No. 146392068840I. DEFINITIONS

[0091] A “B cell” is a lymphocyte that matures within the bone marrow, and includes a naive B cell, memory B cell, or effector B cell (plasma cells). The B cell herein may be a normal or non-malignant B cell.

[0092] The “CD20” antigen, or “CD20,” is an about 35-kDa, non-glycosylated phosphoprotein found on the surface of greater than 90% of B cells from peripheral blood or lymphoid organs. CD20 is present on both normal B cells as well as malignant B cells, but is not expressed on stem cells. Other names for CD20 in the literature include “B -lymphocyte- restricted antigen” and “Bp35”. The CD20 antigen is described in Clark et al. Proc. Natl. Acad. Sci. (USA) 82:1766 (1985), for example.

[0093] “Antibodies” are usually heterotetrameric glycoproteins of about 150,000 daltons, composed of two identical light (L) chains and two identical heavy (H) chains.

[0094] The “light chains” of antibodies (immunoglobulins) from mammalian species can be assigned to one of two clearly distinct types, called kappa (K) and lambda (1), based on the amino acid sequences of their constant domains.

[0095] The “heavy chains” of antibodies from mammalian species can also be assigned to different classes. There are five major classes of intact antibodies: IgA, IgD, IgE, IgG, and IgM, and several of these may be further divided into subclasses (isotypes), e.g., IgGl, IgG2, IgG3, IgG4, IgA, and IgA2. The heavy chain constant domains that correspond to the different classes of antibodies are called a, 6, e, y, and p, respectively. The subunit structures and three-dimensional configurations of different classes of immunoglobulins are well known.

[0096] The term “ocrelizumab” “OCR” (CAS Registration No. 637334-45-3) herein refers to the genetically engineered humanized monoclonal antibody directed against the CD20 antigen and comprising (a) a light chain comprising the amino acid sequence of SEQ ID NO: 9 and (b) a heavy chain comprising the amino acid sequence of SEQ ID NO: 10 or 11. Ocrelizumab® is sold by Genentech®.

[0097] A “subject” or “patient” herein is a human subject or patient. Generally, the subject or patient is eligible for treatment for multiple sclerosis. For the purposes herein, such eligible subject or patient is one who is experiencing, has experienced, or is likely to experience, one or more signs, symptoms or other indicators of multiple sclerosis; has been diagnosed with multiple sclerosis, whether, for example, newly diagnosed (with “new onset” MS), previously13MF-363724269Attorney Docket No. 146392068840 diagnosed with a new relapse or exacerbation, previously diagnosed and in remission, etc.', and / or is at risk for developing multiple sclerosis.

[0098] As used herein, “treatment” or “treating” is an approach for obtaining beneficial or desired results including clinical results. For purposes of this application, beneficial or desired clinical results include, but are not limited to, one or more of the following: decreasing one or more symptoms resulting from the disease, diminishing the extent of the disease, stabilizing the disease (e.g., preventing or delaying the worsening of the disease), delaying or slowing the progression of the disease, ameliorating the disease state, decreasing the dose of one or more other medications required to treat the disease, and / or increasing the quality of life.

[0099] As used herein, “delaying” or “slowing” the progression of multiple sclerosis means to prevent, defer, hinder, slow, retard, stabilize, and / or postpone development of the disease. This delay can be of varying lengths of time, depending on the history of the disease and / or individual being treated.

[0100] A “symptom” of MS is any morbid phenomenon or departure from the normal in structure, function, or sensation, experienced by the subject and indicative of MS.

[0101] “Multiple sclerosis” refers to the chronic inflammatory, often disabling disease of the central nervous system characterized by demyelination and neurodegeneration. There are three internationally recognized forms of MS, namely, primary progressive multiple sclerosis (PPMS), relapsing-remitting multiple sclerosis (RRMS), and secondary progressive multiple sclerosis (SPMS).

[0102] “Progressive multiple sclerosis” as used herein refers to primary progressive multiple sclerosis (PPMS), and secondary progressive multiple sclerosis (SPMS). In some embodiments, progressive multiple sclerosis is characterized by documented, irreversible loss of neurological function persisting for > 6 months that cannot be attributed to clinical relapse.

[0103] “Primary progressive multiple sclerosis” or “PPMS” is characterized by a gradual progression of the disease from its onset with rare superimposed relapses and remissions. There may be periods of a leveling off of disease activity and there may be good and bad days or weeks. PPMS differs from RRMS and SPMS in that onset is typically in the late thirties or early forties, men are as likely women to develop it, and initial disease activity is often in the spinal cord and not in the brain. PPMS disease activity can also be observed (or found) in the brain. PPMS is the sub-type of MS that is least likely to show inflammatory14MF-363724269Attorney Docket No. 146392068840(gadolinium enhancing) lesions on MRI scans. The Primary Progressive form of the disease affects about 15% of all people with multiple sclerosis. PPMS may be defined according to the criteria in Thompson et al. (2018) Lancet 7(2): 162-173. The subject with PPMS treated herein is usually one with probable or definitive diagnosis of PPMS.

[0104] “Relapsing-remitting multiple sclerosis” or “RRMS” is characterized by relapses (also known as exacerbations) during which time new symptoms can appear and old ones resurface or worsen. The relapses are followed by periods of remission, during which time the person fully or partially recovers from the deficits acquired during the relapse. Relapses can last for days, weeks or months and recovery can be slow and gradual or almost instantaneous. The vast majority (about 85%) of people presenting with MS are first diagnosed with RRMS. This is typically when they are in their twenties or thirties, though diagnoses much earlier or later are known. Twice as many women as men present with this sub-type of MS. During relapses, myelin, a protective insulating sheath around the nerve fibers (neurons) in the white matter regions of the central nervous system (CNS), may be damaged in an inflammatory response by the body’s own immune system. This causes a wide variety of neurological symptoms that vary considerably depending on which areas of the CNS are damaged.

[0105] In some embodiments, an initial or first “antibody dose” refers to contact with or exposure to the antibody herein in one or more administrations over a period of time of about 1-20 days. The administrations may be given at one time or at fixed or irregular time intervals over this period of exposure. Initial and later (e.g. second or third) antibody doses are separated in time from each other as described in detail herein. In some embodiments, an initial or first “antibody dose” refers to subcutaneous administration.

[0106] As used herein, an “interval” between antibody doses refers to time period between an earlier antibody dose and a later antibody dose.

[0107] A “stable” formulation is one in which the protein therein essentially retains its physical stability and / or chemical stability and / or biological activity upon storage. Preferably, the formulation essentially retains its physical and chemical stability, as well as its biological activity upon storage. The storage period is generally selected based on the intended shelf life of the formulation. Various analytical techniques for measuring protein stability are available in the art and are reviewed 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), for example. Stability can be measured at a selected temperature for a15MF-363724269Attorney Docket No. 146392068840 selected time period. Stability can be evaluated qualitatively and / or quantitatively in a variety of different ways, including evaluation of aggregate formation (for example using size exclusion chromatography, by measuring turbidity, and / or by visual inspection); by assessing charge heterogeneity using cation exchange chromatography, image capillary isoelectric focusing (icIEF) or capillary zone electrophoresis; amino-terminal or carboxy-terminal sequence analysis; mass spectrometric analysis; SDS-PAGE analysis to compare reduced and intact antibody; peptide map (for example tryptic or LYS-C) analysis; evaluating biological activity or antigen binding function of the antibody; etc. Instability may involve any one or more of: aggregation, deamidation (e.g., Asn deamidation), oxidation (e.g., Met oxidation), isomerization (e.g., Asp isomeriation), clipping / hydrolysis / fragmentation (e.g., hinge region fragmentation), succinimide formation, unpaired cysteine(s), N-terminal extension, C- terminal processing, glycosylation differences, etc.

[0108] As used herein, “biological activity” of a monoclonal antibody includes the ability of the antibody to bind to antigen and resulting in a measurable biological response which can be measured in vitro or in vivo.

[0109] “Corticosteroid” refers to any one of several synthetic or naturally occurring substances with the general chemical structure of steroids that mimic or augment the effects of the naturally occurring corticosteroids. Examples of synthetic corticosteroids include prednisone, prednisolone (including methylprednisolone), dexamethasone, glucocorticoid and betamethasone.

[0110] A “package insert” is used to refer to instructions customarily included in commercial packages of therapeutic products, that contain information about the indications, usage, dosage, administration, contraindications, other therapeutic products to be combined with the packaged product, and / or warnings relating to the use of such therapeutic products, etc.

[0111] “Methionine” described herein refers to L-Methionine.

[0112] A “label” is used herein to refer to information customarily included with commercial packages of pharmaceutical formulations including containers such as vials and package inserts, as well as other types of packaging.

[0113] Reference to “about” a value or parameter herein includes (and describes) variations that are directed to that value or parameter per se. For example, description referring to “about X” includes description of “X.” In some embodiments, “about” a value or parameter describes a range of 50% to 150%, 60% to 140%, 70% to 130%, 80% to 120%, 90% to16MF-363724269Attorney Docket No. 146392068840110%, or 95% to 105% of the value or parameter. In some embodiments, “about” a value or parameter describes a range of 90% to 110% of the value or parameter.

[0114] As used herein and in the appended claims, the singular forms “a,” “or,” and “the” include plural referents unless the context clearly dictates otherwise. It is understood that aspects and variations of the application described herein include “consisting” and / or “consisting essentially of’ aspects and variations.

[0115] The pharmaceutical compositions of the application may include a “therapeutically effective amount” of an anti-CD20 antibody. A “therapeutically effective amount” refers to an amount effective, at dosages and for periods of time necessary, to achieve the desired therapeutic result. A therapeutically effective amount of the antibody may vary according to factors such as the disease state, age, sex, and weight of the individual, and the ability of the antibody or antibody portion to elicit a desired response in the individual. A therapeutically effective amount is also one in which any toxic or detrimental effects of the antibody or antibody portion are outweighed by the therapeutically beneficial effects. Dosage regimens may be adjusted to provide the optimum desired response.

[0116] It is to be understood that one, some, or all of the properties of the various embodiments described herein may be combined to form other embodiments of the present application. These and other aspects of the application are apparent to one of skill in the art.

[0117] All references cited herein, including patent applications and publications, are incorporated by reference in their entirety.II. Antibody Formulations and Preparation

[0118] The application herein relates to aqueous pharmaceutical formulations comprising an anti-CD20 antibody.

[0119] Provided herein is an aqueous pharmaceutical formulation comprising an anti-CD20 antibody in a concentration of about 99 mg / ml to 121 mg / ml (e.g., 110 mg / ml), hyaluronidase in a concentration of at least about 500 U / ml, polyvinylpyrrolidone (PVP) KI 2 in a concentration of about 5% (w / v) to about 10% (w / v), methionine in a concentration of about 10 mM to about 50 mM, sucrose or trehalose, a surfactant, and a buffer, wherein the formulation has a pH of about 5.0 to about 5.6 (e.g., about 5.3), wherein the anti-CD20 antibody comprises a VH domain comprising the amino acid sequence set forth in SEQ ID NO: 8, a VL domain comprising the amino acid sequence set forth in SEQ ID NO: 7, and a human IgGl constant region, wherein the formulation is suitable for subcutaneous17MF-363724269Attorney Docket No. 146392068840 administration. In some embodiments, hyaluronidase is in a concentration of about 1000 U / ml to about 3000 U / ml. In some embodiments, hyaluronidase is in a concentration of about 1500 U / ml to about 2500 U / ml. In some embodiments, hyaluronidase is in a concentration of about 1800 U / ml to about 2200 U / ml. In some embodiments, hyaluronidase is in a concentration of about 2000 U / ml. In some embodiments, the PVP K12 has a weight average molecular weight of less than 7,000 Da. In some embodiments, the PVP K12 has a weight average molecular weight of about 2,000 Da to about 7,000 Da. In some embodiments, the PVP K12 has a weight average molecular weight of about 2,000 Da to about 5,000 Da. In some embodiments, the PVP K12 has a weight average molecular weight of about 3,000 Da to about 7,000 Da. In some embodiments, the PVP KI 2 has a weight average molecular weight of about 5,000 Da to about 7,000 Da. In some embodiments, the PVP KI 2 has a weight average molecular weight of about 2,000 Da to about 3,000 Da. In some embodiments, the PVP K12 is in a concentration of about 5% (w / v) to 7.5% (w / v). In some embodiments, the PVP K12 is in a concentration of 7.5% (w / v). In some embodiments, methionine is in a concentration of about 10 mM to about 40 mM. In some embodiments, methionine is in a concentration of 25 mM to 35 mM. In some embodiments, methionine is in a concentration of 30 mM. In some embodiments, the formulation comprises sucrose. In some embodiments, the sucrose is in a concentration of about 130 to about 260 mM, about 150 mM to about 210 mM. or about 180 mM. In some embodiments, the surfactant is polysorbate 20. In some embodiments, the polysorbate 20 is in a concentration of at least 0.03% (w / v). In some embodiments, the polysorbate is in a concentration of about 0.03% (w / v) to about 0.08% (w / v), about 0.04% (w / v) to about 0.08% (w / v), or about 0.06% (w / v). In some embodiments, the buffer in the formulation is an acetate buffer. In some embodiments, the acetate buffer has an acetate concentration of about 20 mM to about 50 mM. In some embodiments, the acetate buffer has an acetate concentration of about 37 mM to about 47 mM. In some embodiments, the acetate buffer has an acetate concentration of about 43 mM. In some embodiments, the buffer is a sodium acetate buffer. In some embodiments, the sodium acetate buffer has a sodium concentration of about 2 mM to about 8 mM and has an acetate concentration of about 37 mM to about 47 mM. In some embodiments, the sodium acetate buffer has a sodium concentration of about 5mM and an acetate concentration of about 43 mM. In some embodiments, the formulation has a pH of 5.3.18MF-363724269Attorney Docket No. 146392068840

[0120] Further provided herein is an aqueous pharmaceutical formulation comprising an anti- CD20 antibody in a concentration of about 99 mg / ml to 121 mg / ml (e.g., 110 mg / ml), hyaluronidase in a concentration of at least about 500 U / ml, polyvinylpyrrolidone (PVP) having a weight average molecular weight of about 2,000 Da to about 11,000 Da in a concentration of about 5% (w / v) to about 10% (w / v), methionine in a concentration of about 10 mM to about 50 mM, sucrose or trehalose, a surfactant, and a buffer, wherein the formulation has a pH of about 5.0 to about 5.6 (e.g., about 5.3), wherein the anti-CD20 antibody comprises a VH domain comprising the amino acid sequence set forth in SEQ ID NO: 8, a VL domain comprising the amino acid sequence set forth in SEQ ID NO: 7, and a human IgGl constant region, wherein the formulation is suitable for subcutaneous administration. In some embodiments, the PVP is in a concentration of about 5% (w / v) to 7.5% (w / v). In some embodiments, the PVP is in a concentration of 7.5% (w / v). In some embodiments, methionine is in a concentration of about 10 mM to about 40 mM. In some embodiments, methionine is in a concentration of 25 mM to 35 mM. In some embodiments, methionine is in a concentration of 30 mM. In some embodiments, the formulation comprises sucrose. In some embodiments, the sucrose is in a concentration of about 130 to about 260 mM, about 150 mM to about 210 mM. or about 180 mM. In some embodiments, the surfactant is polysorbate 20. In some embodiments, the polysorbate 20 is in a concentration of at least 0.03% (w / v). In some embodiments, the polysorbate is in a concentration of about 0.03% (w / v) to about 0.08% (w / v), about 0.04% (w / v) to about 0.08% (w / v), or about 0.06% (w / v). In some embodiments, the buffer in the formulation is an acetate buffer. In some embodiments, the acetate buffer has an acetate concentration of about 20 mM to about 50 mM. In some embodiments, the acetate buffer has an acetate concentration of about 37 mM to about 47 mM. In some embodiments, the acetate buffer has an acetate concentration of about 43 mM. In some embodiments, the buffer is a sodium acetate buffer. In some embodiments, the sodium acetate buffer has a sodium concentration of about 2 mM to about 8 mM and has an acetate concentration of about 37 mM to about 47 mM. In some embodiments, the sodium acetate buffer has a sodium concentration of about 5mM and an acetate concentration of about 43 mM. In some embodiments, hyaluronidase is in a concentration of about 1000 U / ml to about 3000 U / ml. In some embodiments, hyaluronidase is in a concentration of about 1500 U / ml to about 2500 U / ml. In some embodiments, hyaluronidase is in a concentration of about 1800 U / ml to about 2200 U / ml. In some embodiments, hyaluronidase is in a concentration of about 2000 U / ml. In some embodiments, the formulation has a pH of 5.3.19MF-363724269Attorney Docket No. 146392068840

[0121] Further provided herein is an aqueous pharmaceutical formulation comprising an anti- CD20 antibody in a concentration of about 99 mg / ml to 121 mg / ml (e.g., 110 mg / ml), hyaluronidase in a concentration of at least about 500 U / ml, polyvinylpyrrolidone (PVP) having a weight average molecular weight that is less than 7,000 Da in a concentration of about 5% (w / v) to about 10% (w / v), methionine in a concentration of about 10 mM to about 50 mM, sucrose or trehalose, a surfactant, and a buffer, wherein the formulation has a pH of about 5.0 to about 5.6 (e.g., about 5.3), wherein the anti-CD20 antibody comprises a VH domain comprising the amino acid sequence set forth in SEQ ID NO: 8, a VL domain comprising the amino acid sequence set forth in SEQ ID NO: 7, and a human IgGl constant region, wherein the formulation is suitable for subcutaneous administration. In some embodiments, the PVP is in a concentration of about 5% (w / v) to 7.5% (w / v). In some embodiments, the PVP is in a concentration of 7.5% (w / v). In some embodiments, the PVP has a weight average MW of 2,000 Da to 7,000 Da. In some embodiments, the PVP has a weight average MW of 2,000 Da to 3,000 Da. In some embodiments, the PVP has a weight average MW of 2,000 Da to 5,000 Da. In some embodiments, the PVP has a weight average MW of 5,000 Da to 7,000 Da. In some embodiments, methionine is in a concentration of about 10 mM to about 40 mM. In some embodiments, methionine is in a concentration of 25 mM to 35 mM. In some embodiments, methionine is in a concentration of 30 mM. In some embodiments, the formulation comprises sucrose. In some embodiments, the sucrose is in a concentration of about 130 to about 260 mM, about 150 mM to about 210 mM. or about 180 mM. In some embodiments, the surfactant is polysorbate 20. In some embodiments, the polysorbate 20 is in a concentration of at least 0.03% (w / v). In some embodiments, the polysorbate is in a concentration of about 0.03% (w / v) to about 0.08% (w / v), about 0.04% (w / v) to about 0.08% (w / v), or about 0.06% (w / v). In some embodiments, the buffer in the formulation is an acetate buffer. In some embodiments, the acetate buffer has an acetate concentration of about 20 mM to about 50 mM. In some embodiments, the acetate buffer has an acetate concentration of about 37 mM to about 47 mM. In some embodiments, the acetate buffer has an acetate concentration of about 43 mM. In some embodiments, the buffer is a sodium acetate buffer. In some embodiments, the sodium acetate buffer has a sodium concentration of about 2 mM to about 8 mM and has an acetate concentration of about 37 mM to about 47 mM. In some embodiments, the sodium acetate buffer has a sodium concentration of about 5mM and an acetate concentration of about 43 mM. In some embodiments, hyaluronidase is in a concentration of about 1000 U / ml to about 3000 U / ml. In some embodiments, hyaluronidase is in a concentration of about 1500 U / ml to about 250020MF-363724269Attorney Docket No. 146392068840U / ml. In some embodiments, hyaluronidase is in a concentration of about 1800 U / ml to about 2200 U / ml. In some embodiments, hyaluronidase is in a concentration of about 2000 U / ml. In some embodiments, the formulation has a pH of 5.3.

[0122] Further provided herein is an aqueous pharmaceutical formulation comprising an anti- CD20 antibody in a concentration of about 110 mg / ml to 120 mg / ml , hyaluronidase in a concentration of about 2000 U / ml, polyvinylpyrrolidone (PVP) K12 in a concentration of about 7.5% (w / v), methionine (e.g., at a concentration of 20-30 mM), a stabilizer (e.g., a sugar), a surfactant, and a buffer, wherein the formulation has a pH of about 5.0 to about 5.6 (e.g., about 5.3), wherein the anti-CD20 antibody comprises a VH domain comprising the amino acid sequence set forth in SEQ ID NO: 8, a VL domain comprising the amino acid sequence set forth in SEQ ID NO: 7, and a human IgGl constant region, wherein the formulation is suitable for subcutaneous administration. In some embodiments, the PVP K12 has a weight average molecular weight of about 2,000 Da to about 7,000 Da. In some embodiments, the PVP K12 has a weight average molecular weight of about 2,000 Da to about 5,000 Da, 3,000 Da to about 7,000 Da, 5,000 Da to about 7,000 Da or about 2,000 Da to about 3,000 Da. In some embodiments, the PVP KI 2 has a weight average molecular weight of about 2,000 Da to about 3,000 Da. In some embodiments, methionine is in a concentration of about 10 mM to about 50 mM or 10 mM to about 40 mM. In some embodiments, methionine is in a concentration of 25 mM to 35 mM. In some embodiments, methionine is in a concentration of 30 mM. In some embodiments, the stabilizer is sucrose. In some embodiments, the sucrose is in a concentration of about 130 to about 260 mM, about 150 mM to about 210 mM. or about 180 mM. In some embodiments, the surfactant is polysorbate 20. In some embodiments, the polysorbate 20 is in a concentration of at least 0.03% (w / v). In some embodiments, the polysorbate is in a concentration of about 0.03% (w / v) to about 0.08% (w / v), about 0.04% (w / v) to about 0.08% (w / v), or about 0.06% (w / v). In some embodiments, the buffer in the formulation is an acetate buffer. In some embodiments, the acetate buffer has an acetate concentration of about 20 mM to about 50 mM. In some embodiments, the acetate buffer has an acetate concentration of about 37 mM to about 47 mM. In some embodiments, the acetate buffer has an acetate concentration of about 43 mM. In some embodiments, the buffer is a sodium acetate buffer. In some embodiments, the sodium acetate buffer has a sodium concentration of about 2 mM to about 8 mM and has an acetate concentration of about 37 mM to about 47 mM. In some embodiments, the sodium acetate buffer has a sodium concentration of about 5mM and an21MF-363724269Attorney Docket No. 146392068840 acetate concentration of about 43 mM. In some embodiments, the formulation has a pH of 5.3.

[0123] Provided herein is an aqueous pharmaceutical formulation comprising an anti-CD20 antibody in a concentration of about 99 mg / ml to 121 mg / ml (e.g., 110 mg / ml), hyaluronidase in a concentration of about 1800 U / ml to about 2200 U / ml, polyvinylpyrrolidone (PVP) K12 in a concentration of about 6.75% (w / v) to about 8.25% (w / v), methionine in a concentration of about 27 mM to about 33 mM, sucrose in a concentration of about 162 mM to about 198 mM, a polysorbate 20 in a concentration of about 0.054% (w / v) to about 0.066 (w / v), and an acetate buffer, wherein the formulation has a pH of about 5.0 to about 5.6 (e.g., about 5.3), wherein the anti-CD20 antibody comprises a VH domain comprising the amino acid sequence set forth in SEQ ID NO: 8, a VL domain comprising the amino acid sequence set forth in SEQ ID NO: 7, and a human IgGl constant region, wherein the formulation is suitable for subcutaneous administration. In some embodiments, hyaluronidase is in a concentration of about 2000 U / ml. In some embodiments, the PVP K12 has a weight average molecular weight of less than 7,000 Da. In some embodiments, the PVP K12 has a weight average molecular weight of about 2,000 Da to about 7,000 Da. In some embodiments, the PVP K12 has a weight average molecular weight of about 2,000 Da to about 5,000 Da. In some embodiments, the PVP K12 has a weight average molecular weight of about 3,000 Da to about 7,000 Da. In some embodiments, the PVP K12 has a weight average molecular weight of about 5,000 Da to about 7,000 Da. In some embodiments, the PVP K12 has a weight average molecular weight of about 2,000 Da to about 3,000 Da. In some embodiments, the PVP K12 is in a concentration of 7.5% (w / v). In some embodiments, methionine is in a concentration of 30 mM. In some embodiments, the sucrose is in a concentration of about 180 mM. In some embodiments, the polysorbate 20 is in a concentration of about 0.06% (w / v). In some embodiments, the acetate buffer has an acetate concentration of about 20 mM to about 50 mM. In some embodiments, the acetate buffer has an acetate concentration of about 37 mM to about 47 mM. In some embodiments, the acetate buffer has an acetate concentration of about 43 mM. In some embodiments, the buffer is a sodium acetate buffer. In some embodiments, the sodium acetate buffer has a sodium concentration of about 2 mM to about 8 mM and has an acetate concentration of about 37 mM to about 47 mM. In some embodiments, the sodium acetate buffer has a sodium concentration of about 5mM and an acetate concentration of about 43 mM. In some embodiments, the formulation has a pH of 5.3.22MF-363724269Attorney Docket No. 146392068840

[0124] Provided herein is an aqueous pharmaceutical formulation comprising an anti-CD20 antibody in a concentration of about 104.5 mg / ml to 115.5 mg / ml (e.g., 110 mg / ml), hyaluronidase in a concentration of about 1900 U / ml to about 2100 U / ml, polyvinylpyrrolidone (PVP) KI 2 in a concentration of about 7.125% (w / v) to about 7.875% (w / v), methionine in a concentration of about 28.5 mM to about 31.5 mM, sucrose in a concentration of about 171 mM to about 189 mM, a polysorbate 20 in a concentration of about 0.057% (w / v) to about 0.063 (w / v), and an acetate buffer, wherein the formulation has a pH of about 5.0 to about 5.6 (e.g., about 5.3), wherein the anti-CD20 antibody comprises a VH domain comprising the amino acid sequence set forth in SEQ ID NO: 8, a VL domain comprising the amino acid sequence set forth in SEQ ID NO: 7, and a human IgGl constant region, wherein the formulation is suitable for subcutaneous administration. In some embodiments, hyaluronidase is in a concentration of about 2000 U / ml. In some embodiments, the PVP K12 has a weight average molecular weight of less than 7,000 Da. In some embodiments, the PVP K12 has a weight average molecular weight of about 2,000 Da to about 7,000 Da. In some embodiments, the PVP K12 has a weight average molecular weight of about 2,000 Da to about 5,000 Da. In some embodiments, the PVP K12 has a weight average molecular weight of about 3,000 Da to about 7,000 Da. In some embodiments, the PVP KI 2 has a weight average molecular weight of about 5,000 Da to about 7,000 Da. In some embodiments, the PVP K12 has a weight average molecular weight of about 2,000 Da to about 3,000 Da. In some embodiments, the PVP K12 is in a concentration of 7.5% (w / v). In some embodiments, methionine is in a concentration of 30 mM. In some embodiments, the sucrose is in a concentration of about 180 mM. In some embodiments, the polysorbate 20 is in a concentration of about 0.06% (w / v). In some embodiments, the acetate buffer has an acetate concentration of about 20 mM to about 50 mM. In some embodiments, the acetate buffer has an acetate concentration of about 37 mM to about 47 mM. In some embodiments, the acetate buffer has an acetate concentration of about 43 mM. In some embodiments, the buffer is a sodium acetate buffer. In some embodiments, the sodium acetate buffer has a sodium concentration of about 2 mM to about 8 mM and has an acetate concentration of about 37 mM to about 47 mM. In some embodiments, the sodium acetate buffer has a sodium concentration of about 5mM and an acetate concentration of about 43 mM. In some embodiments, the formulation has a pH of 5.3.

[0125] Further provided herein is an aqueous pharmaceutical formulation comprising an anti- CD20 antibody in a concentration of about 110 mg / ml to 120 mg / ml , hyaluronidase in a23MF-363724269Attorney Docket No. 146392068840 concentration of about 2000 U / ml, polyvinylpyrrolidone (PVP) having a weight average molecular weight that is less than 7,000 Da in a concentration of about 7.5% (w / v), methionine (e.g., at a concentration of 20-30 mM), a stabilizer (e.g., a sugar), a surfactant, and a buffer, wherein the formulation has a pH of about 5.0 to about 5.6 (e.g., about 5.3), wherein the anti-CD20 antibody comprises a VH domain comprising the amino acid sequence set forth in SEQ ID NO: 8, a VL domain comprising the amino acid sequence set forth in SEQ ID NO: 7, and a human IgGl constant region, wherein the formulation is suitable for subcutaneous administration. In some embodiments, the PVP has a weight average molecular weight of about 2,000 Da to about 7,000 Da. In some embodiments, the PVP has a weight average molecular weight of about 2,000 Da to about 5,000 Da, 3,000 Da to about 7,000 Da, 5,000 Da to about 7,000 Da or about 2,000 Da to about 3,000 Da. In some embodiments, the PVP has a weight average molecular weight of about 2,000 Da to about 3,000 Da. In some embodiments, methionine is in a concentration of about 10 mM to about 50 mM or 10 mM to about 40 mM. In some embodiments, methionine is in a concentration of 25 mM to 35 mM. In some embodiments, methionine is in a concentration of 30 mM. In some embodiments, the stabilizer is sucrose. In some embodiments, the sucrose is in a concentration of about 130 to about 260 mM, about 150 mM to about 210 mM. or about 180 mM. In some embodiments, the surfactant is polysorbate 20. In some embodiments, the polysorbate 20 is in a concentration of at least 0.03% (w / v). In some embodiments, the polysorbate is in a concentration of about 0.03% (w / v) to about 0.08% (w / v), about 0.04% (w / v) to about 0.08% (w / v), or about 0.06% (w / v). In some embodiments, the buffer in the formulation is an acetate buffer. In some embodiments, the acetate buffer has an acetate concentration of about 20 mM to about 50 mM. In some embodiments, the acetate buffer has an acetate concentration of about 37 mM to about 47 mM. In some embodiments, the acetate buffer has an acetate concentration of about 43 mM. In some embodiments, the buffer is a sodium acetate buffer. In some embodiments, the sodium acetate buffer has a sodium concentration of about 2 mM to about 8 mM and has an acetate concentration of about 37 mM to about 47 mM. In some embodiments, the sodium acetate buffer has a sodium concentration of about 5mM and an acetate concentration of about 43 mM. In some embodiments, the formulation has a pH of 5.3.

[0126] Further provided herein is an aqueous pharmaceutical formulation comprising an anti- CD20 antibody in a concentration of about 110 mg / ml to 120 mg / ml , hyaluronidase in a concentration of about 2000 U / ml, polyvinylpyrrolidone (PVP) having K- value of 10-14 in a24MF-363724269Attorney Docket No. 146392068840 concentration of about 7.5% (w / v), methionine (e.g., at a concentration of 20-30 mM), a stabilizer e.g., a sugar), a surfactant, and a buffer, wherein the formulation has a pH of about 5.0 to about 5.6 (e.g., about 5.3), wherein the anti-CD20 antibody comprises a VH domain comprising the amino acid sequence set forth in SEQ ID NO: 8, a VL domain comprising the amino acid sequence set forth in SEQ ID NO: 7, and a human IgGl constant region, wherein the formulation is suitable for subcutaneous administration. In some embodiments, the PVP has a weight average molecular weight of about 2,000 Da to about 7,000 Da. In some embodiments, the PVP has a weight average molecular weight of about 2,000 Da to about 5,000 Da, 3,000 Da to about 7,000 Da, 5,000 Da to about 7,000 Da or about 2,000 Da to about 3,000 Da. In some embodiments, the PVP =has a weight average molecular weight of about 2,000 Da to about 3,000 Da. In some embodiments, methionine is in a concentration of about 10 mM to about 50 mM or 10 mM to about 40 mM. In some embodiments, methionine is in a concentration of 25 mM to 35 mM. In some embodiments, methionine is in a concentration of 30 mM. In some embodiments, the stabilizer is sucrose. In some embodiments, the sucrose is in a concentration of about 130 to about 260 mM, about 150 mM to about 210 mM. or about 180 mM. In some embodiments, the surfactant is polysorbate 20. In some embodiments, the polysorbate 20 is in a concentration of at least 0.03% (w / v). In some embodiments, the polysorbate is in a concentration of about 0.03% (w / v) to about 0.08% (w / v), about 0.04% (w / v) to about 0.08% (w / v), or about 0.06% (w / v). In some embodiments, the buffer in the formulation is an acetate buffer. In some embodiments, the acetate buffer has an acetate concentration of about 20 mM to about 50 mM. In some embodiments, the acetate buffer has an acetate concentration of about 37 mM to about 47 mM. In some embodiments, the acetate buffer has an acetate concentration of about 43 mM. In some embodiments, the buffer is a sodium acetate buffer. In some embodiments, the sodium acetate buffer has a sodium concentration of about 2 mM to about 8 mM and has an acetate concentration of about 37 mM to about 47 mM. In some embodiments, the sodium acetate buffer has a sodium concentration of about 5mM and an acetate concentration of about 43 mM. In some embodiments, the formulation has a pH of 5.3.

[0127] Further provided herein is an aqueous pharmaceutical formulation comprising an anti- CD20 antibody in a concentration of about 110 mg / ml to 120 mg / ml, hyaluronidase in a concentration of about 2000 U / ml, polyvinylpyrrolidone (PVP) K12 in a concentration of about 5% (w / v) to about 10% (w / v), methionine in a concentration of about 10 mM to about 50 mM, sucrose having a concentration of about 180 mM to about 240 mM, a surfactant25MF-363724269Attorney Docket No. 146392068840 comprising polysorbate 20 at a concentration of about 0.06% (w / v), and a buffer, wherein the formulation has a pH of about 5.0 to about 5.6 (e.g., about 5.3), wherein the anti-CD20 antibody comprises a VH domain comprising the amino acid sequence set forth in SEQ ID NO: 8, a VL domain comprising the amino acid sequence set forth in SEQ ID NO: 7, and a human IgGl constant region, wherein the formulation is suitable for subcutaneous administration. In some embodiments, methionine in a concentration of about 30 mM. In some embodiments, the buffer in the formulation is an acetate buffer. In some embodiments, the acetate buffer has an acetate concentration of about 20 mM to about 50 mM. In some embodiments, the acetate buffer has an acetate concentration of about 37 mM to about 47 mM. In some embodiments, the acetate buffer has an acetate concentration of about 43 mM. In some embodiments, the buffer is a sodium acetate buffer. In some embodiments, the sodium acetate buffer has a sodium concentration of about 2 mM to about 8 mM and has an acetate concentration of about 37 mM to about 47 mM. In some embodiments, the sodium acetate buffer has a sodium concentration of about 5mM and an acetate concentration of about 43 mM. In some embodiments, the formulation has a pH of 5.3.

[0128] Further provided herein is an aqueous pharmaceutical formulation comprising an anti- CD20 antibody in a concentration of about 110 mg / ml to about 120 mg / ml, hyaluronidase in a concentration of about 2000 U / ml, polyvinylpyrrolidone (PVP) KI 2 in a concentration of about 7.5% (w / v), methionine in a concentration of about 30 mM, sucrose having a concentration of about 180 mM to about 240 mM, a surfactant comprising polysorbate 20 at a concentration of about 0.06% (w / v), and a buffer, wherein the formulation has a pH of about 5.0 to about 5.6 (e.g., about 5.3), wherein the anti-CD20 antibody comprises a VH domain comprising the amino acid sequence set forth in SEQ ID NO: 8, a VL domain comprising the amino acid sequence set forth in SEQ ID NO: 7, and a human IgGl constant region, wherein the formulation is suitable for subcutaneous administration. In some embodiments, the buffer in the formulation is an acetate buffer. In some embodiments, the acetate buffer has an acetate concentration of about 20 mM to about 50 mM. In some embodiments, the acetate buffer has an acetate concentration of about 37 mM to about 47 mM. In some embodiments, the acetate buffer has an acetate concentration of about 43 mM. In some embodiments, the buffer is a sodium acetate buffer. In some embodiments, the sodium acetate buffer has a sodium concentration of about 2 mM to about 8 mM and has an acetate concentration of about 37 mM to about 47 mM. In some embodiments, the sodium acetate buffer has a sodium26MF-363724269Attorney Docket No. 146392068840 concentration of about 5mM and an acetate concentration of about 43 mM. In some embodiments, the formulation has a pH of 5.3.

[0129] Further provided herein is an aqueous pharmaceutical formulation comprising an anti- CD20 antibody in a concentration of about 110 mg / ml, hyaluronidase in a concentration of about 2000 U / ml, polyvinylpyrrolidone (PVP) K12 in a concentration of about 7.5% (w / v), methionine in a concentration of about 30 mM, sucrose having a concentration of about 180 mM, a surfactant comprising polysorbate 20 at a concentration of about 0.06% (w / v), and a buffer, wherein the formulation has a pH of about 5.0 to about 5.6 (e.g., about 5.3), wherein the anti-CD20 antibody comprises a VH domain comprising the amino acid sequence set forth in SEQ ID NO: 8, a VL domain comprising the amino acid sequence set forth in SEQ ID NO: 7, and a human IgGl constant region, wherein the formulation is suitable for subcutaneous administration. In some embodiments, the pH of the formulation is about 5.3. In some embodiments, the buffer in the formulation is an acetate buffer. In some embodiments, the acetate buffer has an acetate concentration of about 20 mM to about 50 mM. In some embodiments, the acetate buffer has an acetate concentration of about 37 mM to about 47 mM. In some embodiments, the acetate buffer has an acetate concentration of about 43 mM. In some embodiments, the buffer is a sodium acetate buffer. In some embodiments, the sodium acetate buffer has a sodium concentration of about 2 mM to about 8 mM and has an acetate concentration of about 37 mM to about 47 mM. In some embodiments, the sodium acetate buffer has a sodium concentration of about 5mM and an acetate concentration of about 43 mM. In some embodiments, the formulation has a pH of 5.3.

[0130] Further provided herein is an aqueous pharmaceutical formulation comprising an anti-CD20 antibody in a concentration of about 100 mg / ml to about 120 mg / ml (e.g., 110 mg / ml), sodium acetate in a concentration of about 20 mM to about 30 mM, sucrose in a concentration of about 180 mM to about 240 mM, methionine in a concentration of about 25 mM to about 30 mM, polysorbate (e.g., polysorbate 20) in a concentration of about 0.06% (w / v), hyaluronidase in a concentration of about or at least about 1500 U / ml (e.g., about or at least about any of 1500 U / ml, 1600 U / ml, 1700 U / ml, 1800 U / ml, and 2000 U / ml), and a low molecular weight PVP (e.g., a PVP with a weight average MW of about 3K to 4K, e.g., PVP K12) in a concentration of about 7.5% (w / v), wherein the formulation has a pH of about 5.0- 5.6 (e.g., about 5.3), wherein the anti-CD20 antibody comprises a VH domain comprising the amino acid sequence set forth in SEQ ID NO: 8, a VL domain comprising the amino acid sequence set forth in SEQ ID NO: 7, and a human IgGl constant region, wherein the27MF-363724269Attorney Docket No. 146392068840 formulation is suitable for subcutaneous administration. In some embodiments, the anti-CD20 antibody comprises a light chain comprising the amino acid sequence of SEQ ID NO: 9 and a heavy chain comprising the amino acid sequence of SEQ ID NO: 10 or 11. In some embodiments, the CD20 antibody is ocrelizumab. In some embodiments, the formulation comprises glacial acetic acid (e.g., at a concentration of about 3.5 mM to about 4.5 mM). In some embodiments, the formulation does not comprise trehalose or trehalose dihydrate. In some embodiments, the sucrose is in a concentration of about 180 mM, about 190 mM, about 200 mM, about 210 mM, about 220 mM, about 230 mM, or about 240 mM. In some embodiments, the sucrose is in a concentration of about 180 mM to about 190 mM, about 190 mM to about 200 mM, about 200 mM to about 210 mM, about 210 mM to about 220 mM, about 220 mM to about 230 mM, about 230 mM to about 240 mM. In some embodiments, the formulation has a pH of 5.3.

[0131] Further provided herein is an aqueous pharmaceutical formulation comprising an anti- CD20 antibody in a concentration of about 110 mg / ml to about 120 mg / ml, sodium acetate in a concentration of about 20 mM, sucrose in a concentration of about 180 mM to about 240 mM, methionine in a concentration of about 30 mM, polysorbate 20 in a concentration of about 0.05% (w / v) to about 0.07 (w / v) (e.g., 0.06% (w / v)), hyaluronidase in a concentration of about 2000 U / ml, and polyvinylpyrrolidone (PVP) K12 in a concentration of about 7.5% (w / v), wherein the formulation has a pH of about 5.3, wherein the anti-CD20 antibody comprises a VH domain comprising the amino acid sequence set forth in SEQ ID NO: 8, a VL domain comprising the amino acid sequence set forth in SEQ ID NO: 7, and a human IgGl constant region, wherein the formulation is suitable for subcutaneous administration. In some embodiments, the anti-CD20 antibody comprises a light chain comprising the amino acid sequence of SEQ ID NO: 9 and a heavy chain comprising the amino acid sequence of SEQ ID NO: 10 or 11. In some embodiments, the CD20 antibody is ocrelizumab. In some embodiments, the formulation comprises glacial acetic acid (e.g., at a concentration of about 3.5 mM to about 4.5 mM). In some embodiments, the sucrose is in a concentration of about 180 mM, about 190 mM, about 200 mM, about 210 mM, about 220 mM, about 230 mM, or about 240 mM. In some embodiments, the sucrose is in a concentration of about 180 mM to about 190 mM, about 190 mM to about 200 mM, about 200 mM to about 210 mM, about 210 mM to about 220 mM, about 220 mM to about 230 mM, about 230 mM to about 240 mM. In some embodiments, the formulation has a pH of 5.3.28MF-363724269Attorney Docket No. 146392068840

[0132] Further provided herein is an aqueous pharmaceutical formulation comprising an anti- CD20 antibody in a concentration of about 110 mg / ml to about 120 mg / ml (e.g., 110 mg / ml), sodium acetate in a concentration of about 20 mM, sucrose in a concentration of about 180 mM to about 240 mM, methionine in a concentration of about 30 mM, polysorbate (e.g., polysorbate 20) in a concentration of about 0.06% (w / v), hyaluronidase in a concentration of about 2000 U / ml, and polyvinylpyrrolidone (PVP) K12 in a concentration of about 7.5% (w / v), wherein the formulation has a pH of about 5.3, wherein the anti-CD20 antibody comprises a VH domain comprising the amino acid sequence set forth in SEQ ID NO: 8, a VL domain comprising the amino acid sequence set forth in SEQ ID NO: 7, and a human IgGl constant region, wherein the formulation is suitable for subcutaneous administration. In some embodiments, the anti-CD20 antibody comprises a light chain comprising the amino acid sequence of SEQ ID NO: 9 and a heavy chain comprising the amino acid sequence of SEQ ID NO: 10 or 11. In some embodiments, the CD20 antibody is ocrelizumab. In some embodiments, the formulation comprises glacial acetic acid (e.g., at a concentration of about 3.5 mM to about 4.5 mM). In some embodiments, the sucrose is in a concentration of about 180 mM, about 190 mM, about 200 mM, about 210 mM, about 220 mM, about 230 mM, or about 240 mM. In some embodiments, the sucrose is in a concentration of about 180 mM to about 190 mM, about 190 mM to about 200 mM, about 200 mM to about 210 mM, about 210 mM to about 220 mM, about 220 mM to about 230 mM, about 230 mM to about 240 mM. In some embodiments, the formulation has a pH of 5.3.

[0133] In some embodiments, the anti-CD20 antibody in the formulation is in an amount of about 100 mg / ml to about 120 mg / ml. In some embodiments, the anti-CD20 antibody in the formulation is in an amount of about 110 mg / ml to about 120 mg / ml. In some embodiments, the buffer is sodium acetate. In some embodiments, the buffer in the formulation is in a concentration of about 15 mM to about 30 mM. In some embodiments, the sodium acetate is in a concentration of about 20 mM to 30 mM. In some embodiments, the sodium acetate is in a concentration of about 20 mM. In some embodiments, the sucrose in the formulation is about 180 mM to about 240 mM. In some embodiments, the sucrose in the formulation is about 180 mM. In some embodiments, the sucrose in the formulation is about 240 mM. In some embodiments, the surfactant in the formulation is polysorbate (e.g., polysorbate 20). In some embodiments, polysorbate 20 in the formulation is in a concentration of about 0.004% (w / v) to about 0.08% (w / v). In some embodiments, polysorbate 20 in the formulation is in a concentration of about 0.06% (w / v). In some embodiments, the formulation has a pH of29MF-363724269Attorney Docket No. 146392068840 about 5.0 to about 6.3. In some embodiments, the formulation has a pH of about 5.0 to about 5.6. In some embodiments, the formulation has a pH of about 5.3. In some embodiments, provided herein is an aqueous pharmaceutical formulation comprising an anti-CD20 antibody in a concentration of about 110 mg / ml to about 120 mg / ml, sodium acetate in a concentration of about 20 mM, sucrose in a concentration of about 180 mM, methionine in a concentration of about 30 mM, polysorbate in a concentration of about 0.06% (w / v), hyaluronidase in a concentration of about 2000 U / ml, and polyvinylpyrrolidone (PVP) KI 2 in a concentration of about 7.5% (w / v), wherein the formulation has a pH of about 5.3. In some embodiments, the formulation is suitable for subcutaneous administration. In some embodiments, the sucrose is in a concentration of about 180 mM, about 190 mM, about 200 mM, about 210 mM, about 220 mM, about 230 mM, or about 240 mM. In some embodiments, the sucrose is in a concentration of about 180 mM to about 190 mM, about 190 mM to about 200 mM, about 200 mM to about 210 mM, about 210 mM to about 220 mM, about 220 mM to about 230 mM, about 230 mM to about 240 mM. In some embodiments, the formulation has a pH of 5.3.

[0134] In some embodiments, the formulation comprises an anti-CD20 antibody in a concentration of about 100 mg / ml to about 120 mg / ml. In some embodiments, the formulation comprises an anti-CD20 antibody in a concentration of about 110 mg / ml to about 120 mg / ml. In some embodiments, the formulation comprises an anti-CD20 antibody in a concentration of about 100 mg / ml to about 110 mg / ml. In some embodiments, the formulation comprises an anti-CD20 antibody in a concentration of about 105 mg / ml to about 115 mg / ml. In some embodiments, the formulation comprises an anti-CD20 antibody in a concentration of about 100 mg / ml, about 105 mg / ml, about 110 mg / ml, about 115 mg / ml, or about 120 mg / ml. In some embodiments, the formulation comprises an anti-CD20 antibody in a concentration of about 100 mg / ml. In some embodiments, the formulation comprises an anti-CD20 antibody in a concentration of about 105 mg / ml. In some embodiments, the formulation comprises an anti-CD20 antibody in a concentration of about 110 mg / ml. In some embodiments, the formulation comprises an anti-CD20 antibody in a concentration of about 115 mg / ml. In some embodiments, the formulation comprises an anti-CD20 antibody in a concentration of about 120 mg / ml. In some embodiments, the anti-CD20 antibody is ocrelizumab.

[0135] In some embodiments, the antibody in the formulation is stable at -20°C for at least about 6 months, at least about 12 months, at least about 18 months, at least two years, at least30MF-363724269Attorney Docket No. 146392068840 three years, or at least four years. In some embodiments, the antibody in the formulation is stable at 2-8°C for at least about 6 months, at least about 12 months, at least about 18 months, or at least 24 months

[0136] In some embodiments, the antibody in the formulation retains at least about any of 50%, 60%, 70%, 80%, 90%, and 95% of its biological activity exhibited before storage, i.e., at the time the pharmaceutical formulation was prepared. In some embodiments, the biological activity is measured by antibody binding to CD20.

[0137] The various aspects and embodiments described in this section in the context of a liquid pharmaceutical composition also apply to articles of manufacture containing such a formulation, in particular, wherein the formulation is in a single fixed dose of the anti-CD20 antibody. In other aspects, the various aspects and embodiments described in this section in the context of a liquid pharmaceutical composition also apply to methods of treating multiple sclerosis in a patient comprising subcutaneously administering an anti-CD20 antibody to the patient, wherein the anti-CD20 antibody is in an aqueous formulation as described herein. Similarly, the various aspects and embodiments described in this section in the context of a liquid pharmaceutical composition also apply to an anti-CD20 antibody for use with a delivery device, according to the methods described herein, and an anti-CD20 antibody for use according to the methods described herein, in combination with a delivery device, and an anti-CD20 antibody for use according to the methods described herein, wherein the anti- CD20 antibody is administered by a delivery device (e.g., an on-body device).

[0138] In some embodiments, the formulation (e.g. the formulation comprising polyvinylpyrrolidone (PVP) K12 in a concentration of about 7.5% (w / v)) has improved biodistribution after SC administration compared to a corresponding formulation comprising polyvinylpyrrolidone (PVP) K12 in a concentration of about 5% or about 10%.

[0139] In some embodiments, the formulation (e.g. the formulation comprising polyvinylpyrrolidone (PVP) K12 in a concentration of about 7.5% (w / v)) has improved biodistribution after SC administration in an animal model (e.g., rat) compared to a corresponding formulation comprising polyvinylpyrrolidone (PVP) KI 2 in a concentration of about 5% or about 10%. In some embodiments, improved biodistribution is measured by percent bolus retention, specifically decreased percent bolus retention, at the subcutaneous injection site, e.g., by quantitative SPEC-CT imaging.31MF-363724269Attorney Docket No. 146392068840

[0140] In some embodiments, the formulation (e.g. the formulation comprising polyvinylpyrrolidone (PVP) K12 in a concentration of about 7.5% (w / v)) has increased systemic exposure of ocrelizumab after SC administration in an animal model e.g., rat) compared to a corresponding formulation comprising polyvinylpyrrolidone (PVP) KI 2 in a concentration of about 5% or about 10%. In some embodiments, the increased systemic exposure of ocrelizumab in a subject is measured via radioactivity measurement in plasma after administration of the liquid pharmaceutical formulation as described herein. In some embodiments, non-compartmental analysis method is used to determine pharmacokinetic parameters based on plasma concentration-time profiles. In some embodiments, improved biodistribution is measured by increased pharmacokinetic Cmax or plasma drug concentrationtime curve (AUC) compared to subjects subcutaneously dosed with a corresponding liquid pharmaceutical formulation.

[0141] In some embodiments, the AUCo- / achieved by the formulation (e.g., the formulation comprising polyvinylpyrrolidone (PVP) K12 in a concentration of about 7.5% (w / v), e.g., the formulation shown in Table 1) after SC administration is increased by at least 5%, 10%, 15%, 20%, or 25% as compared to a corresponding AUCo- / achieved by a control formulation (e.g., a corresponding formulation comprising a 5% or 10% PVP P12).

[0142] In some embodiments, the Cmax achieved by the formulation (e.g., the formulation comprising polyvinylpyrrolidone (PVP) K12 in a concentration of about 7.5% (w / v), e.g., the formulation shown in Table 1) after SC administration is increased by at least 5%, 10%, 15%, 20%, 25% or 28% as compared to a corresponding Cmax achieved by a control formulation (e.g., a corresponding formulation comprising a 5% or 10% PVP P12).

[0143] In some embodiments, the formulation (e.g. the formulation comprising polyvinylpyrrolidone (PVP) K12 in a concentration of about 7.5% (w / v), e.g., the formulation shown in Table 1) achieves a bioavailability after SC administration of at least about any of about 50%, 55%, 60%, 65%, or 70%.Antibodies and their Production

[0144] In some embodiments, the anti-CD20 antibody comprises a humanized anti-CD20 antibody. In some embodiments, the humanized anti-CD20 antibody comprises a light chain variable region comprising CDR LI having the amino acid sequence set forth in SEQ ID NO: 1, CDR L2 having the amino acid sequence set forth in SEQ ID NO: 2, and CDR L3 having the amino acid sequence set forth in SEQ ID NO: 3, and a heavy chain variable region32MF-363724269Attorney Docket No. 146392068840 comprising CDR Hl having the amino acid sequence set forth in SEQ ID NO: 4, CDR H2 having the amino acid sequence set forth in SEQ ID NO: 5, and CDR H3 having the amino acid sequence set forth in SEQ ID NO: 6, and optionally a human IgGl constant region.

[0145] In some embodiments, the anti-CD20 antibody comprises a VH domain comprising the amino acid sequence set forth in SEQ ID NO: 8, a VL domain comprising the amino acid sequence set forth in SEQ ID NO: 7, and a human IgGl constant region.

[0146] In some embodiments, the anti-CD20 antibody comprises a light chain comprising the amino acid sequence of SEQ ID NO: 9 and a heavy chain comprising the amino acid sequence of SEQ ID NO: 10 or 11. In some embodiments, the CD20 antibody is ocrelizumab.

[0147] In some embodiments, the anti-CD20 antibody comprises a light chain comprising the amino acid sequence set forth in SEQ ID NO: 9, and a heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 10, and a human IgGl constant region.

[0148] In some embodiments, the anti-CD20 antibody comprises a light chain comprising the amino acid sequence set forth in SEQ ID NO: 9, and a heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 11, and a human IgGl constant region.

[0149] In some embodiments, the anti-CD20 antibody is ocrelizumab.

[0150] In some embodiments, the amino acid lysine at the C-terminus of the heavy chain of the anti-CD20 antibody is absent.

[0151] Methods of producing such an anti-CD20 antibody are described in further detail in, e.g., U.S. Patent No. 7,799,900.Other components in the formulation

[0152] The formulations described herein further comprise other components such as the components described herein.

[0153] In some embodiments, the formulation comprises a sugar or polyol stabilizer. Exemplary sugar or polyol stabilizers include sucrose, trehalose, mannitol, sorbitol, and glycerol, which is to maintain a osmolality of the formulation with the range between 300- 600 mOsmo / kg. See e.g., Usach et al., Adv Ther. 2019 Nov;36(l l):2986-2996. Studies conducted by inventors supported the formulations described herein with sucrose in a concentration of about 130 mM to about 260 mM achieves the optimal osmolality range of 300-600 mOsmo / kg. In some embodiments, the formulation comprises sucrose. In some embodiments, the sucrose is in a concentration of about 130 mM to about 260 mM. In some33MF-363724269Attorney Docket No. 146392068840 embodiments, the sucrose is in a concentration of about 140 mM to about 240 mM. In some embodiments, the sucrose is in a concentration of about 150 mM to about 220 mM. In some embodiments, the sucrose is in a concentration of about 160 mM to about 200 mM. In some embodiments, the sucrose is in a concentration of about 170 mM to about 190 mM. In some embodiments, the sucrose is in a concentration of about any one or more of 130-135 mM, 135-140 mM, 140-145 mM, 145-150 mM, 150-155 mM, 155-160 mM, 160-165 mM, 165- 170 mM, 170-175 mM, 175-180 mM, 180-185 mM, 185-190 mM, 190-195 mM, 195-200 mM, 200-205 mM, 205-210 mM, 210-215 mM, 215-220 mM, 220-225 mM, 225-230 mM, 230-235 mM, 235-240 mM, 240-245 mM, 245-250 mM, 250-255 mM, and / or 255-260 mM. In some embodiments, the sucrose is in a concentration of about 180 mM to about 240 mM.In some embodiments, the sucrose is in a concentration of about 180 mM to about 200 mM.In some embodiments, the sucrose is in a concentration of about 180 mM to about 220 mM.In some embodiments, the sucrose is in a concentration of about 200 mM to about 240 mM.In some embodiments, the sucrose is in a concentration of about 220 mM to about 240 mM.In some embodiments, the sucrose is in a concentration of about 220 mM to about 260 mM.In some embodiments, the sucrose is in a concentration of about 180 mM, about 190 mM, about 200 mM, about 210 mM, about 220 mM, about 230 mM, or about 240 mM. In some embodiments, the sucrose is in a concentration of about 180 mM to about 190 mM, about 190 mM to about 200 mM, about 200 mM to about 210 mM, about 210 mM to about 220 mM, about 220 mM to about 230 mM, about 230 mM to about 240 mM. In some embodiments, the sucrose is in a concentration of about 180 mM. In some embodiments, the sucrose is in a concentration of about 190 mM. In some embodiments, the sucrose is in a concentration of about 200 mM. In some embodiments, the sucrose is in a concentration of about 210 mM. In some embodiments, the sucrose is in a concentration of about 220 mM. In some embodiments, the sucrose is in a concentration of about 230 mM. In some embodiments, the sucrose is in a concentration of about 240 mM.

[0154] In some embodiments, the formulation comprises a surfactant. Exemplary surfactants include polysorbate 20, polysorbate 80 and poloxamer 188. In some embodiments, the formulation comprises polysorbate. In some embodiments, the formulation comprises polysorbate 20. In some embodiments, the polysorbate 20 is in a concentration of at least about 0.03% (w / v) (e.g., about 0.03% (w / v) to about 0.08% (w / v)). In some embodiments, the polysorbate 20 is in a concentration of about 0.04% (w / v) to about 0.08% (w / v). In some embodiments, the polysorbate 20 is in a concentration of about 0.04% (w / v) to about 0.06%34MF-363724269Attorney Docket No. 146392068840(w / v). In some embodiments, the polysorbate 20 is in a concentration of about 0.06% (w / v) to about 0.08% (w / v). In some embodiments, the polysorbate 20 is in a concentration of about 0.04% (w / v) to about 0.05% (w / v). In some embodiments, the polysorbate 20 is in a concentration of about 0.05% (w / v) to about 0.06% (w / v). In some embodiments, the polysorbate 20 is in a concentration of about 0.06% (w / v) to about 0.07% (w / v). In some embodiments, the polysorbate 20 is in a concentration of about 0.07% (w / v) to about 0.08% (w / v). In some embodiments, the polysorbate 20 is in a concentration of about 0.04% (w / v), about 0.05% (w / v), about 0.06% (w / v), about 0.07% (w / v), about 0.08% (w / v). In some embodiments, the polysorbate 20 is in a concentration of about 0.04% (w / v). In some embodiments, the polysorbate 20 is in a concentration of about 0.05% (w / v). In some embodiments, the polysorbate 20 is in a concentration of about 0.06% (w / v). In some embodiments, the polysorbate 20 is in a concentration of about 0.07% (w / v). In some embodiments, the polysorbate 20 is in a concentration of about 0.08% (w / v). In some embodiments, the polysorbate is polysorbate 80. In some embodiments, the polysorbate 80 is in a concentration of at least about 0.03% (w / v). In some embodiments, the polysorbate 80 is in a concentration of about 0.04% (w / v) to about 0.08% (w / v). In some embodiments, the polysorbate is polysorbate 80. In some embodiments, the polysorbate 80 is in a concentration of about 0.04% (w / v) to about 0.05% (w / v), about 0.05% (w / v) to about 0.06% (w / v), about 0.06% (w / v) to about 0.07% (w / v), and / or about 0.07% (w / v) to about 0.08% (w / v).

[0155] In some embodiments, the formulation comprises polyvinylpyrrolidone (PVP) (CAS number 9003-39-8, also named “povidone”, which is used interchangeably herein). See e.g., WO 2010 / 057109, which is incorporated herein by reference by its entirety. Commercially available PVP is available in a variety of viscosity grades according to its K- value. For example, BASF Corporation produce a PVP K12 called Kollidon® 12, having a K-value falling within the range of 10.2 to 13.8 and a PVP K17 called Kollidon® 17 , having a K- value falling within the range of 15.3 to 18.0. In some embodiments, the PVP has a K-value of 10 to 17. In some embodiments, the PVP has a K-value of 10-14. In some embodiments, the PVP has a K-value of 12.

[0156] The K-value of a PVP solution can be determined by measuring its relative viscosity compared to water as measured using a capillary viscometer at 25 °C and calculated as:K value = logz + (c + 1.5c logz)2+ 1.5c log z — cj / (0.15c + 0.003c2)35MF-363724269Attorney Docket No. 146392068840 where c is the weight, on the anhydrous basis, of the specimen tested in each 100.0 mL of solution (g), and z is the viscosity of the sample solution relative to that of water.

[0157] In some embodiments the PVP has a K value determined as falling in the range 10 - 14, or 10.2 - 13.8. The K value may be determined using the above method. In some embodiments K12 refers to a K value falling in the range 10.2 - 13.8.

[0158] PVP can also be defined by reference to its weight average molecular weight. For example, Kollidon® 12 produced by BASF has a weight average molecular weight of 2000- 3000 Daltons (Da). In some embodiments, the PVP has a weight average molecular weight (MW) of about 2K (z.e., 2000) Da to UK Da. In some embodiments, the formulation comprises PVP with a weight average MW of about 2K-3K, 3K-4K, 4K-5K, 5K-6K, 6K-7K, 7K-8K, 8K-9K, 9K-10K, and / or 10K-11K Da. In some embodiments, the formulation comprises polyvinylpyrrolidone (PVP) with a weight molecular weight (MW) (e.g., weight average MW) of about 2K to 5K, or about 3K to 4K, or about 3.5K. In some embodiments, the formulation comprises polyvinylpyrrolidone (PVP) K12. In some embodiments, the formulation comprises polyvinylpyrrolidone (PVP) K12 in a concentration of about 5% to about 10% (w / v). In some embodiments, the formulation comprises polyvinylpyrrolidone (PVP) K12 in a concentration of about 7.5% (w / v). The molecular weight can be measured by methods such as size exclusion chromatographic (SEC) using light scattering detection, a method to determine an absolute molecular weight without the need to apply a reference material.

[0159] In some embodiments, the formulation comprises PVP K12 that has a weight average molecular weight of 2K Da to 7K Da in a concentration of about 5% to about 10% (w / v) (e.g., about 7.5% (w / v)). In some embodiments, the formulation comprises PVP K12 that has a weight average molecular weight of 2K Da to 7K Da in a concentration of about 7.5% (w / v).

[0160] In some embodiments, the formulation comprises PVP K12 that has a weight average molecular weight of 2K Da to 3K Da in a concentration of about 5% to about 10% (w / v).

[0161] In some embodiments, the formulation comprises PVP KI 2 that has a weight average molecular weight of 2K Da to 3K Da in a concentration of about 7.5% (w / v).

[0162] In some embodiments, the PVP K12 is from BASF. In some embodiments, the PVP KI 2 is from Ashland.36MF-363724269Attorney Docket No. 146392068840

[0163] In some embodiments, the formulation comprises hyaluronidase. In some embodiments, hyaluronidase is in a concentration of at least about 500 U / ml. In some embodiments, hyaluronidase is in a concentration of about 1000 U / ml to about 3000 U / ml. In some embodiments, hyaluronidase is in a concentration of about 1500 U / ml to about 2500 U / ml. In some embodiments, hyaluronidase is in a concentration of about 1800 U / ml to about 2200 U / ml. In some embodiments, hyaluronidase is in a concentration of about 2000 U / ml. In some embodiments, the recombinant human hyaluronidase comprises rHuPH20.

[0164] In some embodiments, the formulation has a pH of about 5.0 to about 5.6. In some embodiments, the formulation has a pH of about 5.0, about 5.1, about 5.2, about 5.3, about 5.4, about 5.5, or about 5.6. In some embodiments, the formulation has a pH of about 5.0 to about 5.1, about 5.1 to about 5.2, about 5.2 to about 5.3, about 5.3 to about 5.4, about 5.4 to about 5.5, about 5.5 to about 5.6. In some embodiments, the formulation has a pH of about 5.0. In some embodiments, the formulation has a pH of about 5.1. In some embodiments, the formulation has a pH of about 5.2. In some embodiments, the formulation has a pH of about 5.3. In some embodiments, the formulation has a pH of about 5.4. In some embodiments, the formulation has a pH of about 5.5. In some embodiments, the formulation has a pH of about 5.6.

[0165] Various kinds of buffer can be used to achieve a pH of about 5.0 to about 5.6. Exemplary buffers include an acetate buffer and a histidine buffer. In some embodiments, the formulation comprises an acetate buffer. In some embodiments, the formulation has an acetate concentration of about 20 mM to about 50 mM. In some embodiments, the acetate buffer has a concentration of about any one or more of 17-22 mM, 22-27 mM, 27-32 mM, 32-37 mM, 37-42 mM, 42-47 mM, and / or 47-52 mM. In some embodiments, the acetate buffer has a concentration of about 37 mM to about 47 mM. In some embodiments, the acetate buffer has a concentration of about 43 mM.

[0166] In some embodiments, the acetate buffer is a sodium acetate buffer. In some embodiments, the sodium acetate buffer has a sodium concentration of about 2 mM to about 8 mM and has an acetate concentration of about 37 mM to about 47 mM. In some embodiments, the sodium acetate buffer has a sodium concentration of about 5mM and an acetate concentration of about 43 mM. In some embodiments, the formulation has a sodium acetate buffer having an acetate concentration of about 15 mM to about 30 mM. In some embodiments, the sodium acetate buffer has an acetate concentration of about 15 mM to 30 mM. In some embodiments, the sodium acetate buffer has an acetate concentration of about37MF-363724269Attorney Docket No. 14639206884020 mM to 30 mM. In some embodiments, the sodium acetate buffer has an acetate concentration of about 15 mM to 25 mM. In some embodiments, the sodium acetate buffer has a concentration of about 15 mM, about 20 mM, about 25 mM, or about 30 mM. In some embodiments, the sodium acetate has an acetate concentration of about 15 mM. In some embodiments, the sodium acetate has an acetate concentration of about 20 mM. In some embodiments, the sodium acetate has an acetate concentration of about 25 mM. In some embodiments, the sodium acetate has an acetate concentration of about 30 mM.

[0167] In some embodiments, the acetate concentration is determined using the Cedex Bio HT with acetate analysis.

[0168] In some embodiments, the sodium concentrations is measured using an ion-selective electrode method.

[0169] In some embodiments, the formulation comprises glacial acetic acid. In some embodiments, the formulation comprises glacial acetic acid at a concentration of about 3.5 mM to about 4.5 mM. In some embodiments, the formulation comprises glacial acetic acid at a concentration of about 3.5 mM, 4 mM, 4.5 mM, or 5 mM. In some embodiments, the formulation comprises glacial acetic acid at a concentration of about 3.5 mM to about 4 mM, about 4 mM to about 4.5 mM, or about 4.5 mM to about 5 mM. In some embodiments, the formulation comprises glacial acetic acid at a concentration of about 3.5 mM. In some embodiments, the formulation comprises glacial acetic acid at a concentration of about 4 mM. In some embodiments, the formulation comprises glacial acetic acid at a concentration of about 5 mM.

[0170] In some embodiments, the formulation is essentially free of one or more preservatives, such as benzyl alcohol, phenol, m-cresol, chlorobutanol and benzethonium Cl. In some embodiments, the formulation is essentially free of hydrogen peroxide (e.g., having no more than 500 ppm, 400 ppm, 300 ppm, or 200 ppm).

[0171] In some embodiments, the formulation does not comprise arginine.

[0172] In some embodiments, the physical stability, chemical stability, or biological activity of the antibody in the formulation is evaluated or measured. Any methods known in the art and described in the Examples herein may be used to evaluate the stability and biological activity of the antibody in the formulation. For example, stability of the antibody in the formulation can be measured by, but not limited to, size exclusion chromatography (SEC or SE-HPLC), imaged capillary isoelectric focusing (ICIEF), peptide mapping, small-volume38MF-363724269Attorney Docket No. 146392068840 light obscuration (HIAC) assay, and capillary electrophoresis (CE) techniques such as CE- sodium dodecyl sulfate (CE-SDS) and CE-glycan analysis. In some embodiments, the antibody in the formulation is stable at -20°C for at least about 6 months, at least about 8 months, at least about 10 months, at least about 12 months, at least about 14 months, at least about 16 months, at least about 18 months, at least about 20 months, at least about 21 months, at least about 22 months, at least about 23 months, at least about 24 months, at least about 3 years, or at least about 4 years. In some embodiments, the antibody in the formulation is stable at 2°C to 8°C (e.g., 5°C) for at least about 6 months, at least about 8 months, at least about 10 months, at least about 12 months, at least about 14 months, at least about 16 months, at least about 18 months, at least about 20 months, at least about 21 months, at least about 22 months, at least about 23 months, or at least about 24 months. In some embodiments, the stability of the antibody (i.e., an antibody monomer) is measured by size exclusion chromatography in the formulation after storage. In some embodiments, the stability of the antibody is (i.e., an antibody monomer) measured by imaged capillary isoelectric focusing in the formulation after storage. In some embodiments, the percent of antibody monomer in the formulation as compared to total protein (e.g., including antibody and aggregates) is greater than about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94% or about 95% after storage at -20 °C for at least about 6 months, at least about 12 months, at least about 18 months, or at least about 24 months. In some embodiments, the percent of antibody monomer in the formulation as compared to (e.g., including antibody and aggregates) is greater than about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94% or about 95% after storage at 2°C to 8°C (e.g., 5°C) for at least about 6 months, at least about 12 months, at least about 18 months, or at least about 24 months. In some embodiments, the percent of antibody monomer in the formulation as compared to (e.g., including antibody and aggregates) is greater than about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94% or about 95% after agitation at room temperature (e.g., about 15°C to 25°C) for at least about 2 hours, at least about 4 hours, at least about 6 hours, at least about 8 hours, at least about 10 hours, at least about 12 hours, at least about 14 hours, at least about 16 hours, at least about 18 hours, at least about 20 hours, or at least about 24 hours. In some embodiments, the percent of total aggregates (e.g., high molecular weight species and low molecular weight species) in the formulation is less than39MF-363724269Attorney Docket No. 146392068840 any of about 0.1%, about 0.2%, about 0.3%, about 0.4%, about 0.5%, about 0.6%, about 0.7%, about 0.8%, about 0.9%, about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, or about 10% after storage at -20°C for at least about 6 months, at least about 12 months, at least about 18 months, or at least about 24 months. In some embodiments, the percent of total aggregates (e.g., high molecular weight species and low molecular weight species) in the formulation is less than any of about 0.1%, about 0.2%, about 0.3%, about 0.4%, about 0.5%, about 0.6%, about 0.7%, about 0.8%, about 0.9%, about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, or about 10% after storage at 2°C to 8°C (e.g., 5°C) for at least about 6 months, at least about 12 months, at least about 18 months, or at least about 24 months. In some embodiments, the percent of total aggregates (e.g., high molecular weight species and low molecular weight species) in the formulation is less than any of about 0.1%, about 0.2%, about 0.3%, about 0.4%, about 0.5%, about 0.6%, about 0.7%, about 0.8%, about 0.9%, about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, or about 10% after agitation at room temperature (e.g., about 15°C to 25°C) for at least about 2 hours, at least about 4 hours, at least about 6 hours, at least about 8 hours, at least about 10 hours, at least about 12 hours, at least about 14 hours, at least about 16 hours, at least about 18 hours, at least about 20 hours, or at least about 24 hours.

[0173] The methods for preparing the formulations described herein can be found in the Examples such as Example 1. For example, anti-CD20 antibody formulation can be prepared by concentrating in 20 mM sodium acetate using a tangential flow filtration system which is to achieve a pH of about 5.0-5.6 (e.g., pH of about 5.3). Specifically, anti-CD20 antibody was produced and purified in a 20mM sodium acetate buffer. The purification process includes affinity chromatography, multimodal anion-exchange chromatography and hydrophobic interaction chromatography. The antibody was then subject to a small virus retentive filtration, ultrafiltration and diafiltration (UF / DF). See e.g., W02006031560 for exemplary process for concentrating antibodies, the content of which is incorporated hereby by reference in its entirety. A conditioning buffer with all other ingredients except hyaluronidase is prepared and mixed with the antibody; and hyaluronidase is then added to obtain the final formulation.III. Articles of Manufacture

[0174] The application further provides articles of manufacture or kits (such as kits-of parts) containing a formulation describe herein useful for the treatment of multiple sclerosis. In40MF-363724269Attorney Docket No. 146392068840 some embodiments, the formulation is in a single fixed dose of the anti-CD20 antibody. In some embodiments, the article of manufacture comprises a syringe or a device comprising a needle containing the formulation, optionally wherein the device is an on-body device.

[0175] In some embodiments, the article of manufacture further comprises a label denoting that the anti-CD20 antibody or pharmaceutical formulation is indicated for treating patients with multiple sclerosis (e.g., RMS or PPMS) according to a method described herein and / or is indicated for subcutaneous administration.

[0176] In some embodiments, the article of manufacture or kit comprises, packaged together, a pharmaceutical composition comprising an aqueous pharmaceutical formulation described herein and a label denoting the anti-CD20 antibody or pharmaceutical composition is indicated for treating patients with multiple sclerosis and suppresses disability progression in patients having multiple sclerosis. In some embodiments, the article of manufacture or kit comprises, packaged together, an aqueous pharmaceutical formulation described herein and a label denoting the anti-CD20 antibody or pharmaceutical composition is indicated for treating patients with multiple sclerosis (e.g., RMS or PPMS). In some embodiments, the label provides instructions for administering a therapeutically effective amount of an anti-CD20 antibody subcutaneously to the patient. In some embodiments, the label states that the anti- CD20 antibody is administered subcutaneously every 6 months for about 12, 18, or 24 months. In some embodiments, the label states that the anti-CD20 antibody is administered subcutaneously every 24 weeks for about 12, 18, or 24 months. In some embodiments, the anti-CD20 antibody comprises a VH domain comprising the amino acid set forth in SEQ ID NO: 8, a VL domain comprising the amino acid sequence set forth in SEQ ID NO: 7, and a human IgGl constant region. In some embodiments, the anti-CD20 antibody comprises a light chain comprising the amino acid sequence of SEQ ID NO: 9 and a heavy chain comprising the amino acid sequence of SEQ ID NO: 11.

[0177] In certain embodiments, the article of manufacture or kit comprises a container and a label or package insert on or associated with the container. Suitable containers include, for example, bottles, vials, syringes (e.g., prefilled syringes), etc. The containers may be formed from a variety of materials such as glass or plastic. The container holds or contains a composition that is effective for treating the multiple sclerosis and may have a sterile access port (for example the container may be an intravenous solution bag or a vial having a stopper pierceable by a hypodermic injection needle).41MF-363724269Attorney Docket No. 146392068840

[0178] In some embodiments, the article of manufacture or kit comprises a device suitable for subcutaneous administration comprising a needle. In some embodiments, the device is an on-body injector. In some embodiments, the device comprises an infusion pump. In some embodiments, the device comprises a CRONO ambulatory infusion pump. In some embodiments, the device comprises a Lapas patch pump. In some embodiments, the device comprises an enFuse on-body platform. In some embodiments, the device comprises a wearable injector or wearable injection device. In some embodiments, the device comprises a DrugDeliverySystem. See e.g., Badkar et al., Drug Des Devel Ther. 2021 Jan 13; 15: 159- 170.

[0179] Optionally, the article of manufacture or kit provided herein further comprises a container comprising an agent other than the antibody for treatment and further comprising instructions on treating the patient with such agent.

[0180] In some embodiments, the article of manufacture comprises a single-use vial containing a quantity of an anti-CD20 antibody formulation suitable for subcutaneous administration to a patient, wherein said quantity is sufficient to deliver a therapeutically effective amount of the anti-CD20 antibody. In some embodiments, the anti-CD20 antibody comprises a VH domain comprising the amino acid sequence set forth in SEQ ID NO: 8, a VL domain comprising the amino acid sequence set forth in SEQ ID NO: 7, and a human IgGl constant region. In some embodiments, the anti-CD20 antibody is ocrelizumab.IV. Methods of Treatment.

[0181] The present application further comprises methods of treating multiple sclerosis in a patient comprising subcutaneously administering an anti-CD20 antibody to the patient, wherein the anti-CD20 antibody is in an aqueous pharmaceutical formulation described herein. In some embodiments, the formulation comprises an anti-CD20 antibody in a concentration of about 110 mg / ml to about 120 mg / ml e.g., 110 mg / ml), sodium acetate in a concentration of about 20 mM, sucrose in a concentration of about 180 mM to about 240 mM, methionine in a concentration of about 30 mM, polysorbate in a concentration of about 0.06% (w / v), hyaluronidase in a concentration of about 2000 U / ml, and polyvinylpyrrolidone (PVP) K12 in a concentration of about 7.5% (w / v), and wherein the formulation has a pH of about 5.3, wherein the anti-CD20 antibody comprises a VH domain comprising the amino acid sequence set forth in SEQ ID NO: 8, a VL domain comprising the amino acid sequence set forth in SEQ ID NO: 7, and a human IgGl constant region. In some embodiments, said anti-CD20 antibody in the formulation is about 110 mg / ml. In some embodiments, the anti-42MF-363724269Attorney Docket No. 146392068840CD20 antibody comprises a light chain comprising the amino acid sequence of SEQ ID NO: 9 and a heavy chain comprising the amino acid sequence of SEQ ID NO: 10 or 11. In some embodiments, the anti-CD20 antibody is ocrelizumab.

[0182] In some embodiments, the patient has not been subjected to a prior intravenous administration of the anti-CD20 antibody at a dose of about 600 mg. In some embodiments, the patient has not been subjected to a prior intravenous administration of the anti-CD20 antibody. In some embodiments, the patient has not been subjected to a prior subcutaneous administration of the anti-CD20 antibody at a dose lower than about 900 mg.

[0183] In some embodiments, the patient has received a prior treatment with an anti-CD20 antibody, optionally wherein the prior treatment comprises an intravenous administration of the anti-CD20 antibody, optionally the prior treatment comprises the intravenous administration of the anti-CD20 antibody at a dose of about 300 mg or about 600 mg.

[0184] In some embodiments, the method comprises subcutaneously administering more than one dose of the anti-CD20 antibody. In some embodiments, the method comprises administering at least two, three, or four doses of the anti-CD20 antibody.

[0185] In some embodiments, the anti-CD20 antibody is administered at a frequency of no more than once every 24 weeks or about once every 24 weeks.

[0186] In some embodiments, the anti-CD20 antibody is administered at a frequency of no more than once every 6 months or about once every 6 months.

[0187] In some embodiments, the anti-CD20 antibody is the only medicament administered to the patient to treat multiple sclerosis.

[0188] In some embodiments, the patient is subject to an oral dexamethasone and an oral antihistamine administration shortly prior to the subcutaneous administration of the anti- CD20 antibody, optionally the oral dexamethasone and antihistamine are administered within 30 minutes, 20 minutes, or 15 minutes prior to the subcutaneous administration, further optionally the patient is subject to 20 mg dexamethasone and 5 mg desloratadine.

[0189] In some embodiments, the multiple sclerosis is relapsing multiple sclerosis (RMS).

[0190] In some embodiments, the patient has a clinical isolated syndrome (CIS), relapsingremitting multiple sclerosis (RRMS) or active secondary progressive multiple sclerosis (SPMS).43MF-363724269Attorney Docket No. 146392068840

[0191] In some embodiments, the multiple sclerosis is a primary progressive multiple sclerosis (PPMS).

[0192] In some embodiments, the patient has an Expanded Disability Status Scale (EDSS) score of 0-6.5. In some embodiments, the patient has an Expanded Disability Status Scale (EDSS) score of 0-0.5, 0.5-1, 1-1.5, 1.5-2, 2-2.5, 2.5-3, 3-3.5, 3.5-4, 4-4.5, 4.5-5, 5-5.5, 5.5-6, and / or 6-6.5.

[0193] In some embodiments, the subcutaneous administration takes no more than about any of 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 minutes. In some embodiments the subcutaneous administration uses a needle having a gauge from 25 to 30, or a gauge of about 24 to 26. In some embodiments the administration takes no more than about 30, 25, 20, 15, or 10 minutes using a 24 gauge needle.B. Dosing Regimen

[0194] The following section describes various aspects (embodiments) of dosing and treatment regimens, any and all of which apply to the methods described herein.

[0195] The present application in one aspect provides a method of treating multiple sclerosis in an individual comprising administering an effective amount of an anti-CD20 antibody formulation subcutaneously. In some embodiments, the anti-CD20 antibody is administered every six months or 24 weeks.

[0196] In some embodiments, the anti-CD20 antibody formulation is administered subcutaneously by a healthcare professional to an individual with a manual syringe or syringe pump. In some embodiments, the anti-CD20 antibody formulation is administered through a SC infusion set into the abdominal SC space, except for the 5 cm area directly around the navel, about every six months or 24 weeks. In some embodiments, a minimum of about 22 weeks or about five months is maintained between each subcutaneous dose of anti-CD20 antibody. In some embodiments, the anti-CD20 antibody formulation is administered at home. In some embodiments, the anti-CD20 antibody formulation is administered in a healthcare facility.

[0197] Additional SC spaces may be used for SC administration. These spaces include the outer area of the upper arm, the front of the thigh, midway to the outer side, 4 inches below the top of the thigh to 4 inches above the knee, the upper back, and the lower back in the upper area of the buttock, just behind the hip bone, this has the slowest rate of absorption among sites.44MF-363724269Attorney Docket No. 146392068840

[0198] In some embodiments, provided herein are methods of treating individuals with MS comprising administering a therapeutically effective amount an antibody in the formulation described herein, wherein the antibody binds to CD20 comprising a variable heavy chain (VH) and a variable light chain (VL), wherein the VH comprises a CDRL1 comprising the amino acid sequence set forth in SEQ ID NO: 1, a CDRL2 comprising the amino acid sequence set forth in SEQ ID NO: 2, a CDRL3 comprising the amino acid sequence set forth in SEQ ID NO: 3, a CDRH1 comprising the amino acid sequence set forth in SEQ ID NO: 4, a CDRH2 comprising the amino acid sequence set forth in SEQ ID NO: 5, a CDRH3 comprising the amino acid sequence set forth in SEQ ID NO: 6 to the individual. In some embodiments, the CD20 antibody has a VL domain comprising the amino acid sequence set forth in SEQ ID NO: 7 and a VH domain comprising the amino acid sequence set forth in SEQ ID NO: 8. In some embodiments, the antibody comprises a light chain comprising the amino acid sequence set forth in SEQ ID NO: 9 and a heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 10. In some embodiments, the antibody is ocrelizumab. In some embodiments, the antibody has an IgGl isotype. In some embodiments, the antibody is a humanized antibody. In some embodiments, the antibody is a full-length antibody. In some embodiments, the anti-CD20 antibody formulation is administered about every 6 months or 24 weeks. In some embodiments, the anti-CD20 antibody formulation is administered at least twice, at least three times, at least four times or at least five times. In some embodiments, the anti-CD20 antibody formulation is administered about every six months or about every 24 weeks for about 24 months. In some embodiments the individual has secondary progressive multiple sclerosis (SPMS). In some embodiments the individual has relapsing remitting multiple sclerosis (RRMS). In some embodiments, the individual has relapsing MS (RMS).

[0199] In some embodiments, to reduce potential injection reactions, about 20 mg of dexamethasone (or an equivalent corticosteroid) and / or about 5 mg of desloratadine are administered to the individual prior to subcutaneous administration of an anti-CD20 antibody. In some embodiments, 20 mg of dexamethasone (or an equivalent corticosteroid) and / or 5 mg of desloratadine are administered to the individual shortly before subcutaneous administration of an anti-CD20 antibody. In some embodiments, 20 mg of dexamethasone (or an equivalent corticosteroid) and / or 5 mg of desloratadine are administered to the individual at least about any of 60 minutes, 50 minutes, 40 minutes, or 30 minutes (e.g., at least about 30 minutes) prior to subcutaneous administration of an anti-CD20 antibody. In some embodiments, 20 mg of dexamethasone (or an equivalent corticosteroid) and 5 mg of desloratadine are45MF-363724269Attorney Docket No. 146392068840 administered to the individual about 30 min to about 60 minutes, about 30 minutes to about 40 minutes, about 40 minutes to about 50 minutes, or about 50 minutes to about 60 minutes prior to subcutaneous administration of an anti-CD20 antibody. In some embodiments an equivalent dose of an alternative steroid or antihistaminic is administered to the individual prior to subcutaneous administration of an anti-CD20 antibody.

[0200] The application herein relates to aqueous pharmaceutical formulations comprising an anti-CD20 antibody. In some embodiments, the formulation comprises an anti-CD20 antibody, sodium acetate, sucrose, a buffer, methionine, hyaluronidase, polyvinylpyrrolidone, and polysorbate (e.g., polysorbate 20), wherein the formulation has a pH of about 5.0 to about 7.0. In some embodiments, the anti-CD20 antibody in the formulation is in an amount of about 100 mg / ml to about 120 mg / ml. In some embodiments, the anti-CD20 antibody in the formulation is in an amount of about 110 mg / ml to about 120 mg / ml. In some embodiments, the buffer is sodium acetate. In some embodiments, the buffer in the formulation is in a concentration of about 15 mM to about 30 mM. In some embodiments, the sodium acetate is in a concentration of about 15 mM to 30 mM. In some embodiments, the sodium acetate is in a concentration of about 20 mM. In some embodiments, the sucrose in the formulation is about 180 mM to about 240 mM. In some embodiments, the sucrose in the formulation is about 180 mM. In some embodiments, the sucrose in the formulation is about 240 mM. In some embodiments, the polysorbate in the formulation is polysorbate 20. In some embodiments, polysorbate 20 in the formulation is in a concentration of about 0.04% (w / v) to about 0.08% (w / v). In some embodiments, polysorbate 20 in the formulation is in a concentration of about 0.06% (w / v). In some embodiments, the formulation comprises hyaluronidase in a concentration of about 2000 U / ml. In some embodiments, the formulation comprises polyvinylpyrrolidone (PVP) K12 in a concentration of about 7.5% (w / v). In some embodiments, the formulation has a pH of about 5.0 to about 6.3. In some embodiments, the formulation has a pH of about 5.0 to about 5.6. In some embodiments, the formulation has a pH of about 5.3. In some embodiments, provided herein is an aqueous pharmaceutical formulation comprising an anti-CD20 antibody in a concentration of about 110 mg / ml to about 120 mg / ml, sodium acetate in a concentration of about 20 mM, sucrose in a concentration of about 180 mM to about 240 mM, methionine in a concentration of about 30 mM, polysorbate 20 in a concentration of about 0.06% (w / v), hyaluronidase in a concentration of about 2000 U / ml, and polyvinylpyrrolidone (PVP) KI 2 in a concentration of about 7.5% (w / v), wherein the formulation has a pH of about 5.3. In some embodiments, the46MF-363724269Attorney Docket No. 146392068840 formulation is suitable for subcutaneous administration. In some embodiments, the recombinant human hyaluronidase comprises rHuPH20. In some embodiments, the anti- CD20 antibody is ocrelizumab.

[0201] In some embodiments, an anti-CD20 antibody in the aqueous pharmaceutical formulation described herein is administered multiple times (e.g., at least two times, or at least three times) over a period of 12 months or more. In some embodiments, an anti-CD20 antibody is administered multiple times (e.g., at least two, three, four times) over a period of 18 months or more. In some embodiments, an anti-CD20 antibody is administered multiple times (e.g., at least two, three, four or five times) over a period of 24 months or more.

[0202] In some aspects, provided herein is a method of treating MS in an individual comprising administering an anti-CD20 antibody in the aqueous pharmaceutical formulation described herein subcutaneously to the individual about every six months or 24 weeks for about at least 12 months, at least 18 months, or at least 24 months; wherein each administration of the anti-CD20 antibody comprises a subcutaneous administration of a therapeutically effective amount of an anti-CD20 antibody.

[0203] In some embodiments, the method comprises (i) administering an anti-CD20 antibody in the aqueous pharmaceutical formulation described herein subcutaneously to the individual on day 0, (ii) administering a second dose of the anti-CD20 antibody in the aqueous pharmaceutical formulation described herein subcutaneously about 6 months (e.g., 180, 181, 182, 183, 184, or 185 days) later, (iii) administering a third dose of the anti-CD20 antibody in the aqueous pharmaceutical formulation described herein subcutaneously about 12 months later, (iv) administering a fourth dose of the anti-CD20 antibody in the aqueous pharmaceutical formulation described herein subcutaneously about 18 months later, (v) administering a fifth dose of the anti-CD20 antibody in the aqueous pharmaceutical formulation described herein subcutaneously about 24 months later. In some embodiments, a therapeutically effective amount of the anti-CD20 antibody in the aqueous pharmaceutical formulation described herein is administered during each dose.

[0204] In some embodiments, the anti-CD20 antibody is the only medicament administered to the individual to treat multiple sclerosis. In some embodiments, the anti-CD20 antibody is the only disease modifying therapy (DMT) administered to the patient to treat multiple sclerosis. For example, in some embodiments, the anti-CD20 antibody is administered in47MF-363724269Attorney Docket No. 146392068840 combination with one or more of: methylprednisolone (or equivalent); an antihistamine (e.g., diphenhydramine or equivalent); an analgesic e.g., acetaminophen); and an antipyretic.

[0205] In some embodiments, the anti-CD20 antibody in the aqueous pharmaceutical formulation described herein is administered via a device suitable for subcutaneous administration. In some embodiments, the device comprises an infusion pump. In some embodiments, the device comprises a CRONO ambulatory infusion pump. In some embodiments, the device comprises a Lapas patch pump. In some embodiments, the device comprises an enFuse® on-body platform. In some embodiments, the device comprises a wearable injector or wearable injection device. In some embodiments, the device comprises a drug delivery system. See e.g., Badkar et al., Drug Des Devel Ther. 2021 Jan 13; 15: 159- 170.

[0206] In some embodiments, provided herein are methods of treating individuals with multiple sclerosis by administering an anti-CD20 antibody subcutaneously to the individual, wherein each subcutaneous anti-CD20 antibody administration comprises delivering a therapeutically effective amount of the anti-CD20 antibody SC. In some embodiments the individual has secondary progressive multiple sclerosis (SPMS). In some embodiments the individual has relapsing remitting multiple sclerosis (RRMS). In some embodiments, the individual has relapsing MS (RMS).

[0207] Also provided herein is a method of treating MS in an individual comprising a dosing schedule of at least 24 months comprising administering a therapeutically effective amount of an anti-CD20 antibody in the aqueous pharmaceutical formulation described herein SC about every 6 months or 24 weeks. In some embodiments, consecutive doses of the anti-CD20 antibody is administered at least about 22 weeks or about five months apart.

[0208] In some embodiments, the method comprises administering an anti-CD20 antibody in the aqueous pharmaceutical formulation described herein on weeks 0, 26, 52, 78, and 104. In some embodiments, the method comprises administering an anti-CD20 antibody on weeks 0, 24, 48, 72, and 96. In some embodiments, administering an anti-CD20 antibody comprises a subcutaneous injection by a healthcare professional to an individual with a manual syringe or syringe pump. In some embodiments, subcutaneous administration of an anti-CD20 antibody comprises a therapeutically effective amount of the anti-CD20 antibody.

[0209] In some embodiments, provided herein is a method of treating MS in an individual comprising administering an anti-CD20 antibody in the aqueous pharmaceutical formulation described herein subcutaneously, wherein the anti-CD20 antibody administration comprises48MF-363724269Attorney Docket No. 146392068840 delivering a therapeutically effective amount of the anti-CD20 antibody subcutaneously to the individual during an anti-CD20 antibody delivery period, wherein during the anti-CD20 antibody delivery period, there is a therapeutically effective amount of anti-CD20 antibody in the serum of the individual for treating MS.C. Premedication

[0210] In certain embodiments, the patient is premedicated prior to (e.g., shortly prior to) subcutaneous administration with an anti-CD20 antibody in the aqueous pharmaceutical formulation described herein. Administering one or more premedication into a patient “shortly” prior to the subcutaneous administration in some cases refers to administering the one or more premedication within about 2 hours, 1 hour, 45 minutes, 30 minutes, 20 minutes, 15 minutes, 10 minutes, or 5 minutes prior to the subcutaneous administration of the anti- CD20 antibody.

[0211] In certain embodiments, the patient is premedicated with methylprednisolone (or an equivalent, e.g., an alternative steroid) within approximately 30 minutes to an hour prior to each subcutaneous administration of anti-CD20 antibody in the aqueous pharmaceutical formulation described herein. In certain embodiments, the patient is premedicated with methylprednisolone (or an equivalent) within about 30 minutes e.g., within about 20 minutes or 15 minutes) prior to each subcutaneous administration of anti-CD20 antibody. In certain embodiments, the patient is premedicated with 100 mg IV methylprednisolone (or an equivalent) within approximately 30 minutes to an hour prior to each subcutaneous administration of anti-CD20 antibody. In certain embodiments, the patient is premedicated with 100 mg IV methylprednisolone (or an equivalent) within about 30 minutes (e.g., within about 20 minutes or 15 minutes) prior to each subcutaneous administration of anti-CD20 antibody.

[0212] In certain embodiments, the patient is additionally (or alternatively) premedicated with an antihistaminic drug (e.g. diphenhydramine) approximately 30-60 minutes before each subcutaneous administration of anti-CD20 antibody in the aqueous pharmaceutical formulation described herein. In certain embodiments, the patient is additionally (or alternatively) premedicated with an antihistaminic drug (e.g. diphenhydramine) within about 30 minutes (e.g., within about 20 minutes or 15 minutes) before each subcutaneous administration of anti-CD20 antibody. In certain embodiments, the patient is additionally (or alternatively) premedicated with an antipyretic (e.g. acetaminophen / paracetamol)49MF-363724269Attorney Docket No. 146392068840 approximately 30-60 minutes before each subcutaneous administration of anti-CD20 antibody. In certain embodiments, the patient is additionally (or alternatively) premedicated with an antipyretic (e.g. acetaminophen / paracetamol) within about 30 minutes (e.g., within about 20 minutes or 15 minutes) before each subcutaneous administration of anti-CD20 antibody.D. Prior treatment

[0213] In some embodiments, the patient has been subjected to a prior treatment for MS.

[0214] In some embodiments, the patient has not been subjected to a prior treatment for MS.

[0215] In some embodiments, the patient has not been subjected to a prior intravenous administration of an anti-CD20 antibody (such as any anti-CD20 antibody described herein) at a dose of about 600 mg. In some embodiments, the patient has not been subjected to a prior intravenous administration of an anti-CD20 antibody (such as any anti-CD20 antibody described herein).

[0216] In some embodiments, the patient has not been subjected to a prior subcutaneous administration of an anti-CD20 antibody (such as any anti-CD20 antibody described herein) at a dose lower than about 900 mg.

[0217] In some embodiments, the patient has not been previously administered an anti-CD20 antibody (such as any anti-CD20 antibody described herein). In some embodiments, the patient has not been previously administered an anti-CD20 antibody (such as any anti-CD20 antibody described herein) within 2 years prior to the start of treatment with anti-CD20 antibody described herein. In some embodiments, the anti-CD20 antibody is ocrelizumab.

[0218] In some embodiments, the patient has received a prior treatment with an anti-CD20 antibody. In some embodiments, the prior treatment comprises an intravenous administration of the anti-CD20 antibody. In some embodiments, the prior treatment comprises the intravenous administration of the anti-CD20 antibody at a dose of about 600 mg. In some embodiments, the prior treatment is at least about 1 month, 2 months, 3 months, 6 months, 9 months, 12 months before the time when the methods described herein are conducted.

[0219] In some embodiments, the patient has not been subjected to a prior treatment with cladribine, atacicept, or alemtuzumab.50MF-363724269Attorney Docket No. 146392068840

[0220] In some embodiments, the patient has not been subjected to a prior treatment with fingolimod, siponimod, ponesimod, or ozanimod within 6 weeks prior to the methods comprising subcutaneously administering the anti-CD20 antibody described herein.

[0221] In some embodiments, the patient has not been subjected to a prior treatment with interferons beta (la or lb), or glatiramer acetate within 2 weeks prior to the methods comprising subcutaneously administering the anti-CD20 antibody described herein.

[0222] In some embodiments, the patient has not been subjected to a prior treatment with natalizumab within 4.5 months prior to the methods comprising subcutaneously administering the anti-CD20 antibody described herein.

[0223] In some embodiments, the patient has not been subjected to a prior treatment with mitoxantrone within 2 years prior to the methods comprising subcutaneously administering the anti-CD20 antibody described herein.

[0224] The specification is considered to be sufficient to enable one skilled in the art to practice the application. Various modifications of the application in addition to those shown and described herein will become apparent to those skilled in the art from the foregoing description and fall within the scope of the appended claims. All publications, patents, and patent applications cited herein are hereby incorporated by reference in their entirety for all purposes.EXEMPLARY EMBODIMENTS - PART A

[0225] 1. An aqueous pharmaceutical formulation, the formulation comprising an anti- CD20 antibody in a concentration of about 110 mg / ml to about 120 mg / ml, sodium acetate in a concentration of about 20 mM, sucrose in a concentration of about 180 mM to about 240 mM, methionine in a concentration of about 30 mM, polysorbate in a concentration of about 0.06% (w / v), hyaluronidase in a concentration of about 2000 U / ml, and polyvinylpyrrolidone (PVP) K12 in a concentration of about 7.5% (w / v), and wherein the formulation has a pH of about 5.3, wherein the anti-CD20 antibody comprises a VH domain comprising the amino acid sequence set forth in SEQ ID NO: 8, a VL domain comprising the amino acid sequence set forth in SEQ ID NO: 7, and a human IgGl constant region, wherein the formulation is suitable for subcutaneous administration.

[0226] 2. The formulation of embodiment 1, wherein said anti-CD20 antibody in the formulation is about 110 mg / ml.51MF-363724269Attorney Docket No. 146392068840

[0227] 3. The formulation of embodiment 1 or embodiment 2, wherein the anti-CD20 antibody comprises a light chain comprising the amino acid sequence of SEQ ID NO: 9 and a heavy chain comprising the amino acid sequence of SEQ ID NO: 10 or 11.

[0228] 4. The formulation of any one of embodiments 1-3, wherein the anti-CD20 antibody is ocrelizumab.

[0229] 5. The formulation of any one of embodiments 1-4, wherein the hyaluronidase is recombinant human hyaluronidase.

[0230] 6. The formulation of any one of embodiments 1-5, wherein the formulation is stable at 2-8°C for at least 6 months, at least 12 months, at least 18 months or at least 24 months.

[0231] 7. The formulation of embodiment 6, wherein the antibody in the formulation retains at least about 50%, 60%, 70%, 80%, 90%, or 95% of its biological activity after storage.

[0232] 8. The formulation of embodiment 7, wherein the biological activity is measured by antibody binding to CD20.

[0233] 9. The formulation of any one of embodiment 1-8, which is sterile.

[0234] 10. An article of manufacture containing a formulation of any one of embodiments 1-9, wherein the formulation is in a single fixed dose of the anti-CD20 antibody.

[0235] 11. The article of manufacture of embodiment 10, which comprises a single-used vial.

[0236] 12. The article of manufacture of embodiment 10 or embodiment 11, which comprises a syringe or a device comprising a needle, optionally wherein the device is an on- body device.

[0237] 13. The article of manufacture of any one of embodiments 10-12, further comprising a package insert instructing a user to administer the fixed doses of the anti-CD20 antibody subcutaneously to a patient with multiple sclerosis, optionally wherein the user is the patient.

[0238] 14. A method of treating multiple sclerosis in a patient comprising subcutaneously administering an anti-CD20 antibody to the patient, wherein the anti-CD20 antibody is in an52MF-363724269Attorney Docket No. 146392068840 aqueous formulation comprising an anti-CD20 antibody in a concentration of about 110 mg / ml to about 120 mg / ml, sodium acetate in a concentration of about 20 mM, sucrose in a concentration of about 180 mM, methionine in a concentration of about 30 mM, polysorbate in a concentration of about 0.06% (w / v), hyaluronidase in a concentration of about 2000 U / ml, and polyvinylpyrrolidone (PVP) KI 2 in a concentration of about 7.5% (w / v), and wherein the formulation has a pH of about 5.3, wherein the anti-CD20 antibody comprises a VH domain comprising the amino acid sequence set forth in SEQ ID NO: 8, a VL domain comprising the amino acid sequence set forth in SEQ ID NO: 7, and a human IgGl constant region.

[0239] 15. The method of embodiment 14, wherein the patient has not been subjected to a prior intravenous administration of the anti-CD20 antibody at a dose of about 600 mg.

[0240] 16. The method of embodiment 14 or embodiment 15, wherein the patient has not been subjected to a prior intravenous administration of the anti-CD20 antibody.

[0241] 17. The method of any one of embodiments 14-16, wherein the patient has not been subjected to a prior subcutaneous administration of the anti-CD20 antibody at a dose lower than about 900 mg.

[0242] 18. The method of embodiment 14, wherein the patient has received a prior treatment with an anti-CD20 antibody, optionally wherein the prior treatment comprises an intravenous administration of the anti-CD20 antibody, optionally wherein the prior treatment comprises the intravenous administration of the anti-CD20 antibody at a dose of about 300 mg or about 600 mg.

[0243] 19. The method of any one of embodiments 14-18, wherein the method comprises subcutaneously administering more than one dose of the anti-CD20 antibody.

[0244] 20. The method of embodiment 19, wherein the method comprises administering at least two, three or four doses of the anti-CD20 antibody.

[0245] 21. The method of any one of embodiments 14-20, wherein the anti-CD20 antibody is administered at a frequency of no more than once every 24 weeks or about once every 24 weeks.

[0246] 22. The method of any one of embodiments 14-21, wherein the anti-CD20 antibody is administered at a frequency of no more than once every 6 months or about once every 6 months.53MF-363724269Attorney Docket No. 146392068840

[0247] 23. The method of any one of embodiments 14-22, wherein anti-CD20 antibody is the only medicament administered to the patient to treat multiple sclerosis.

[0248] 24. The method of any one of embodiments 14-23, wherein the patient is subject to an oral dexamethasone and an oral antihistamine administration shortly prior to the subcutaneous administration of the anti-CD20 antibody, optionally wherein the oral dexamethasone and antihistamine are administered within 30 minutes, 20 minutes, or 15 minutes prior to the subcutaneous administration, further optionally wherein the patient is subject to 20 mg dexamethasone and 5 mg desloratadine.

[0249] 25. The method of any one of embodiments 14-24, wherein the multiple sclerosis is relapsing multiple sclerosis (RMS).

[0250] 26. The method of embodiment 14-25, wherein the patient has a clinical isolated syndrome (CIS), relapsing-remitting multiple sclerosis (RRMS) or active secondary progressive multiple sclerosis (SPMS).

[0251] 27. The method of any one of embodiments 14-26, wherein the multiple sclerosis is a primary progressive multiple sclerosis (PPMS).

[0252] 28. The method of any one of embodiments 14-27, wherein the patient has an Expanded Disability Status Scale (EDSS) score of 0-6.5.

[0253] 29. The method of any one of embodiments 14-28, wherein the subcutaneous administration takes no more than about 10 minutes.

[0254] 30. The method of any one of embodiments 14-29, wherein said anti-CD20 antibody in the formulation is about 110 mg / ml.

[0255] 31. The method of any one of embodiments 14-30, wherein the anti-CD20 antibody comprises a light chain comprising the amino acid sequence of SEQ ID NO: 9 and a heavy chain comprising the amino acid sequence of SEQ ID NO: 10 or 11.

[0256] 32. The method of any one of embodiments 14-31, wherein the anti-CD20 antibody is ocrelizumab.

[0257] 33. An aqueous pharmaceutical formulation, the formulation comprising ocrelizumab in a concentration of 110 mg / ml, sodium acetate in a concentration of 20 mM, sucrose in a concentration of 180 mM or 240 mM, methionine in a concentration of 30 mM, polysorbate in a concentration of 0.06% (w / v), hyaluronidase in a concentration of 2000 U / ml, and polyvinylpyrrolidone (PVP) K12 in a concentration of 7.5% (w / v), and wherein the54MF-363724269Attorney Docket No. 146392068840 formulation has a pH of 5.3, wherein the formulation is suitable for subcutaneous administration.

[0258] 34. The formulation of embodiment 33, wherein sucrose is in a concentration of 180 mM.

[0259] 35. The formulation of embodiment 33, wherein sucrose is in a concentration of 240 mM.

[0260] 36. A method of treating multiple sclerosis in a patient comprising subcutaneously administering the formulation of any one of embodiments 33-35.EXEMPLARY EMBODIMENTS - PART B

[0261] 1. An aqueous pharmaceutical formulation, the formulation comprising an anti- CD20 antibody in a concentration of about 99 mg / ml to about 121 mg / ml, hyaluronidase in a concentration of at least about 500 U / ml, polyvinylpyrrolidone (PVP) K12in a concentration of about 5% (w / v) to about 10% (w / v), methionine in a concentration of about 10 mM to about 50 mM, sucrose or trehalose, a surfactant, and a buffer, wherein the formulation has a pH of about 5.0 to about 5.6, wherein the anti-CD20 antibody comprises a VH domain comprising the amino acid sequence set forth in SEQ ID NO: 8, a VL domain comprising the amino acid sequence set forth in SEQ ID NO: 7, and a human IgGl constant region, wherein the formulation is suitable for subcutaneous administration. In some embodiments, hyaluronidase is in a concentration of about 1000 U / ml to about 3000 U / ml. In some embodiments, hyaluronidase is in a concentration of about 1500 U / ml to about 2500 U / ml. In some embodiments, hyaluronidase is in a concentration of about 1800 U / ml to about 2200 U / ml. In some embodiments, hyaluronidase is in a concentration of about 2000 U / ml.

[0262] 2. The formulation of embodiment 1, wherein the anti-CD20 antibody is in a concentration of about 110 mg / ml.

[0263] 3. The formulation of embodiment 1 or embodiment 2, wherein the PVP K12 is in a concentration of about 5% (w / v) to about 7.5% (w / v), optionally wherein the PVP K12 is in a concentration of about 7.5% (w / v).

[0264] 4. The formulation of any one of embodiments 1-3, wherein the hyaluronidase is in a concentration of about 2000 U / ml.55MF-363724269Attorney Docket No. 146392068840

[0265] 5. The formulation of any one of embodiments 1-4, wherein methionine is in a concentration of about 10 mM to about 40 mM, optionally wherein methionine is in a concentration of about 25 mM to about 35 mM, further optionally wherein methionine is in a concentration of about 30 mM.

[0266] 6. The formulation of any one of embodiments 1-5, wherein the formulation comprises sucrose, optionally wherein the sucrose is in a concentration of about 150 mM to about 210 mM.

[0267] 7. The formulation of embodiment 6, wherein the sucrose is in a concentration of about 180 mM.

[0268] 8. The formulation of any one of embodiments 1-7, wherein the surfactant is polysorbate 20.

[0269] 9. The formulation of embodiment 8, wherein the polysorbate is in a concentration of at least about 0.03% (w / v), optionally wherein the polysorbate is in a concentration of about 0.03% (w / v) to about 0.08% (w / v) or about 0.04% (w / v) to about 0.08% (w / v).

[0270] 10. The formulation of embodiment 9, wherein the polysorbate is in a concentration of about 0.06% (w / v).

[0271] 11. The formulation of any one of embodiments 1-10, wherein the pH is about 5.3.

[0272] 12. The formulation of any one of embodiments 1-11, wherein the buffer in the formulation is an acetate buffer.

[0273] 13. The formulation of embodiment 12, wherein the acetate buffer has an acetate concentration of about 20 mM to about 50 mM.

[0274] 14. The formulation of embodiment 13, wherein the acetate buffer has an acetate concentration of about 37 mM to about 47 mM, optionally wherein the acetate buffer has an acetate concentration of about 43 mM.

[0275] 15. The formulation of any one of embodiments 1-14, wherein the buffer is a sodium acetate buffer.

[0276] 16. The formulation of embodiment 15, wherein the sodium acetate buffer has a sodium concentration of about 2 mM to about 8 mM and an acetate concentration of about 37 mM to about 47 mM.56MF-363724269Attorney Docket No. 146392068840

[0277] 17. The formulation of embodiment 16, wherein the sodium acetate buffer has a sodium concentration of about 5 mM and an acetate concentration of about 43 mM.

[0278] 18. An aqueous pharmaceutical formulation, the formulation comprising an anti- CD20 antibody in a concentration of 110 mg / ml, sucrose in a concentration of 180 mM, methionine in a concentration of 30 mM, polysorbate 20 in a concentration of 0.06% (w / v), hyaluronidase in a concentration of 2000 U / ml, polyvinylpyrrolidone (PVP) KI 2 in a concentration of about 7.5% (w / v), and an acetate buffer, and wherein the formulation has a pH of 5.0 to 5.6 (e.g., 5.3), wherein the anti-CD20 antibody comprises a VH domain comprising the amino acid sequence set forth in SEQ ID NO: 8, a VL domain comprising the amino acid sequence set forth in SEQ ID NO: 7, and a human IgGl constant region, wherein the formulation is suitable for subcutaneous administration.

[0279] 19. The formulation of embodiment 18, wherein the formulation has a sodium acetate buffer, and wherein the sodium acetate buffer has a sodium concentration of 5 mM and an acetate concentration of 43 mM.

[0280] 20. The formulation of any one of embodiments 1-19, wherein the weight weight average molecular weight of PVP KI 2 is 2000 Dalton to 7000 Dalton, optionally wherein the weight average molecular weight of PVP K12 is 2000 Dalton to 3000 Dalton.

[0281] 21. The formulation of any one of embodiments 1-20, wherein the anti-CD20 antibody comprises a light chain comprising the amino acid sequence of SEQ ID NO: 9 and a heavy chain comprising the amino acid sequence of SEQ ID NO: 10 or 11.

[0282] 22. The formulation of any one of embodiments 1-21, wherein the anti-CD20 antibody is ocrelizumab.

[0283] 23. The formulation of any one of embodiments 1-22, wherein the hyaluronidase is recombinant human hyaluronidase.

[0284] 24. The formulation of any one of embodiments 1-23, wherein: a) the antibody in the formulation is stable at 2-8°C for at least 6 months, at least 12 months, at least 18 months or at least 24 months, b) the antibody and hyaluronidase in the formulation are stable after continuous agitation for 24 hours at room temperature, and / or c) there is i) less than 1% of aggregation of the antibody in the formulation as assessed by sum of HMW forms by SE- UHPLC after 25 days at 40°C and / or ii) less than 6% of degradation of the antibody in the formulation as measured by sum of pre-peaks by CE-SDS after 25 days at 40°C.57MF-363724269Attorney Docket No. 146392068840

[0285] 25. The formulation of embodiment 24, wherein the hyaluronidase in the formulation is stable when it retains at least about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of its biological activity.

[0286] 26. The formulation of embodiment 24 or embodiment 25, wherein the antibody in the formulation is stable when it retains at least about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of its biological activity.

[0287] 27. The formulation of embodiment 26, wherein the biological activity is measured by antibody binding to CD20.

[0288] 28. The formulation of any one of embodiments 1-27, which is sterile.

[0289] 29. A method of promoting solubility and bioavailability of an anti-CD20 antibody in an aqueous formulation, comprising adding a buffer comprising polyvinylpyrrolidone (PVP) K12 into a solution comprising the anti-CD20 antibody for preparation of the formulation, wherein the formulation comprises the antibody in a concentration of about 100 mg / ml to about 130 mg / ml and the PVP K12 in a concentration of about 7.5% (w / v).

[0290] 30. The method of embodiment 29, wherein the formulation further comprises hyaluronidase.

[0291] 31. A method of protecting an aqueous pharmaceutical formulation comprising an anti-CD20 antibody and hyaluronidase from agitation stress, comprising adding a buffer comprising polyvinylpyrrolidone (PVP) KI 2 into a solution comprising the anti-CD20 antibody for preparation of the formulation, wherein the formulation comprises the PVP K12 in a concentration of at least about 5% (w / v).

[0292] 32. The method of embodiment 31, wherein the PVP K12 has a concentration of between about 5% (w / v) and about 10% (w / v), optionally wherein the PVP K12 has a concentration of about 7.5% (w / v).

[0293] 33. The method of any one of embodiments 30-32, wherein the hyaluronidase is in a concentration of at least 500 U / ml. In some embodiments, hyaluronidase is in a concentration of about 1000 U / ml to about 3000 U / ml. In some embodiments, hyaluronidase is in a concentration of about 1500 U / ml to about 2500 U / ml. In some embodiments, hyaluronidase is in a concentration of about 1800 U / ml to about 2200 U / ml, optionally58MF-363724269Attorney Docket No. 146392068840 wherein the hyaluronidase is in a concentration of about 2000 U / ml, further optionally wherein the hyaluronidase is a recombinant human hyaluronidase.

[0294] 34. The method of any one of embodiments 29-33, wherein the anti-CD20 antibody is in a concentration of about 110 mg / ml.

[0295] 35. The method of any one of embodiments 29-34, wherein the formulation further comprises methionine, optionally wherein methionine is in a concentration of about 10 mM to 50 mM, or about 10 mM to about 40 mM, or about 25 mM to about 35 mM, further optionally wherein methionine is in a concentration of about 30 mM.

[0296] 36. The method of any one of embodiments 29-35, wherein the formulation comprises sucrose or trehalose.

[0297] 37. The method of embodiment 36, wherein the formulation comprises sucrose, optionally wherein the formulation comprises sucrose in a concentration of about 150 mM to about 210 mM, further optionally wherein the sucrose is in a concentration of about 180 mM.

[0298] 38. The method of any one of embodiments 29-37, wherein the surfactant is polysorbate, optionally polysorbate 20.

[0299] 39. The method of embodiment 38, wherein the polysorbate is in a concentration of about 0.04% (w / v) to about 0.08% (w / v), optionally wherein the polysorbate is in a concentration of about 0.06% (w / v).

[0300] 40. The method of any one of embodiments 29-39, wherein the pH is about 4.8 to about 5.8, optionally about 5.0 to about 5.6, further optionally 5.3.

[0301] 41. The method of any one of embodiments 29-40, wherein the formulation comprises a sodium acetate buffer.

[0302] 42. The method of embodiment 41, wherein the sodium acetate buffer has an acetate concentration of about 20 mM to about 50 mM.

[0303] 43. The method of embodiment 41, wherein the acetate concentration is about 37 mM to about 47 mM, further optionally wherein the acetate concentration is about 43 mM.

[0304] 44. The method of embodiment 43, wherein the sodium acetate buffer has a sodium concentration of about 2 mM to about 8 mM, optionally wherein the sodium concentration is about 5 mM.59MF-363724269Attorney Docket No. 146392068840

[0305] 45. The method of any one of embodiments 29-44, wherein the formulation comprises an anti-CD20 antibody in a concentration of about 110 mg / ml, sucrose in a concentration of about 180 mM, methionine in a concentration of about 30 mM, polysorbate 20 in a concentration of about 0.06% (w / v), hyaluronidase in a concentration of about 2000 U / ml, and PVP K12 in a concentration of about 7.5% (w / v), a sodium acetate buffer having a sodium concentration of about 5 mM and an acetate concentration of about 43 mM, and wherein the formulation has a pH of about 5.3.

[0306] 46. The method of any one of embodiments 29-45, wherein the weight average molecular weight of PVP KI 2 is 2000 Dalton to 7000 Dalton, optionally wherein the molecular weight is assessed by size exclusion chromatography (SEC) using light scattering detection, further optionally wherein the weight average molecular weight of PVP KI 2 is 2000 Dalton to 3000 Dalton.

[0307] 47. The method of any one of embodiments 29-46, wherein the anti-CD20 antibody comprises a VH domain comprising the amino acid sequence set forth in SEQ ID NO: 8, a VL domain comprising the amino acid sequence set forth in SEQ ID NO: 7, and a human IgGl constant region, and wherein the formulation is suitable for subcutaneous administration.

[0308] 48. The method of embodiment 47, wherein the anti-CD20 antibody comprises a light chain comprising the amino acid sequence of SEQ ID NO: 9 and a heavy chain comprising the amino acid sequence of SEQ ID NO: 10 or 11.

[0309] 49. A method of treating multiple sclerosis in a patient comprising subcutaneously administering the formulation of any one of embodiments 1-20.

[0310] 50. The method of embodiment 49, wherein the patient has not been subjected to a prior intravenous administration of the anti-CD20 antibody at a dose of about 600 mg.

[0311] 51. The method of embodiment 49 or embodiment 50, wherein the patient has not been subjected to a prior intravenous administration of the anti-CD20 antibody.

[0312] 52. The method of any one of embodiments 49-51, wherein the patient has not been subjected to a prior subcutaneous administration of the anti-CD20 antibody at a dose lower than about 900 mg.

[0313] 53. The method of embodiment 49, wherein the patient has received a prior treatment with an anti-CD20 antibody, optionally wherein the prior treatment comprises an60MF-363724269Attorney Docket No. 146392068840 intravenous administration of the anti-CD20 antibody, optionally wherein the prior treatment comprises the intravenous administration of the anti-CD20 antibody at a dose of about 300 mg or about 600 mg.

[0314] 54. The method of any one of embodiments 49-53, wherein the method comprises subcutaneously administering more than one dose of the anti-CD20 antibody.

[0315] 55. The method of embodiment 54, wherein the method comprises administering at least two, three or four doses of the anti-CD20 antibody.

[0316] 56. The method of any one of embodiments 49-55, wherein the anti-CD20 antibody is administered at a frequency of no more than once every 24 weeks or about once every 24 weeks.

[0317] 57. The method of any one of embodiments 49-56, wherein the anti-CD20 antibody is administered at a frequency of no more than once every 6 months or about once every 6 months.

[0318] 58. The method of any one of embodiments 49-57, wherein anti-CD20 antibody is the only medicament administered to the patient to treat multiple sclerosis.

[0319] 59. The method of any one of embodiments 49-58, wherein the patient is subject to a) an oral dexamethasone or an equivalent corticosteroid and b) an oral antihistamine administration (e.g., desloratadine) shortly prior to the subcutaneous administration of the anti-CD20 antibody, optionally wherein the oral dexamethasone or the equivalent corticosteroid and antihistamine are administered at least or within 30 minutes, 20 minutes, or 15 minutes prior to the subcutaneous administration, further optionally wherein the patient is subject to 20 mg dexamethasone and 5 mg desloratadine.

[0320] 60. The method of any one of embodiments 49-59, wherein the multiple sclerosis is relapsing multiple sclerosis (RMS).

[0321] 61. The method of any one of embodiments 49-60, wherein the patient has a clinical isolated syndrome (CIS), relap sing -remitting multiple sclerosis (RRMS) or active secondary progressive multiple sclerosis (SPMS).

[0322] 62. The method of any one of embodiments 49-61, wherein the multiple sclerosis is a primary progressive multiple sclerosis (PPMS).

[0323] 63. The method of any one of embodiments 49-62, wherein the patient has an Expanded Disability Status Scale (EDSS) score of 0-6.5.61MF-363724269Attorney Docket No. 146392068840

[0324] 64. The method of any one of embodiments 49-63, wherein the subcutaneous administration takes no more than about 10 minutes.

[0325] 65. The method of any one of embodiments 49-64, wherein said anti-CD20 antibody in the formulation is about 110 mg / ml.

[0326] 66. The method of any one of embodiments 49-65, wherein the anti-CD20 antibody comprises a light chain comprising the amino acid sequence of SEQ ID NO: 9 and a heavy chain comprising the amino acid sequence of SEQ ID NO: 10 or 11.

[0327] 67. The method of any one of embodiments 49-66, wherein the anti-CD20 antibody is ocrelizumab.EXEMPLARY EMBODIMENTS - PART C

[0328] The following numbered statements relate to aspects of the present disclosure and form part of the description.

[0329] 1. An aqueous pharmaceutical formulation, the formulation comprising an anti- CD20 antibody in a concentration of about 99 mg / ml to 121 mg / ml, polyvinylpyrrolidone (PVP) K12 in a concentration of about 5% (w / v) to about 10% (w / v), a surfactant, and a buffer, wherein the formulation has a pH of about 5.0 to about 5.6, wherein the anti-CD20 antibody comprises a VH domain comprising the amino acid sequence set forth in SEQ ID NO: 8, a VL domain comprising the amino acid sequence set forth in SEQ ID NO: 7, and a human IgGl constant region, wherein the formulation is suitable for subcutaneous administration.

[0330] 2. The formulation of statement 1, wherein the anti-CD20 antibody is in a concentration of about 100 to about 120 mg / ml, optionally about 110 mg / ml.

[0331] 3. The formulation of statement 1 or statement 2, wherein:(i) the PVP K12 is in a concentration of about 5% (w / v) to 7.5% (w / v),(ii) the PVP K12 is in a concentration of about 7.5% (w / v) to 10% (w / v),(iii) the PVP K12 is in a concentration of 7.0 % to 8.0 %; or(iv) the PVP K12 is in a concentration of 7.5% (w / v).

[0332] 4. The formulation of any one of statements 1-3, wherein the formulation comprises hyaluronidase, wherein the hyaluronidase is optionally in a concentration of about62MF-363724269Attorney Docket No. 1463920688401000 U / ml to about 3000 U / ml, about 1500 U / ml to about 2500 U / ml, or about 1800 U / ml to about 2200 U / ml, optionally about 2000 U / ml.

[0333] 5. The formulation of any one of statements 1-4, wherein the formulation comprises methionine, optionally wherein the methionine is in a concentration of about 10 mM to about 50 mM, of about 20 mM to about 50 mM, of about 30 mM to about 50 mM, or about 30 mM.

[0334] 6. The formulation of any one of statements 1-5, wherein the formulation comprises sucrose or trehalose, wherein the formulation optionally comprises sucrose, further optionally wherein the sucrose is in a concentration of about 150 mM to about 210 mM, further optionally wherein the formulation does not comprise trehalose.

[0335] 7. The formulation of statement 6, wherein the sucrose is in a concentration of about 180 mM.

[0336] 8. The formulation of any one of statements 1-7, wherein the surfactant is polysorbate 20.

[0337] 9. The formulation of statement 8, wherein the polysorbate is in a concentration of at least 0.03% (w / v), optionally wherein the polysorbate is in a concentration of about 0.03% (w / v) to about 0.08% (w / v) or about 0.04% (w / v) to about 0.08% (w / v).

[0338] 10. The formulation of statement 9, wherein the polysorbate is in a concentration of about 0.06% (w / v).

[0339] 11. The formulation of any one of statement 1-10, wherein the pH is about 5.1 - 5.6, or 5.2 - 5.5, or about 5.3.

[0340] 12. The formulation of any one of statements 1-11, wherein the formulation does not comprise arginine.

[0341] 13. The formulation of any one of statements 1-12, wherein the buffer in the formulation is an acetate buffer, wherein optionally the acetate buffer has an acetate concentration of about 20 mM to about 50 mM.

[0342] 14. The formulation of statement 13, wherein the acetate buffer has an acetate concentration of about 37 mM to about 47 mM, optionally wherein the acetate buffer has an acetate concentration of about 43 mM.63MF-363724269Attorney Docket No. 146392068840

[0343] 15. The formulation of any one of statements 1-14, wherein the buffer is a sodium acetate buffer.

[0344] 16. The formulation of statement 15, wherein the sodium acetate buffer has a sodium concentration of about 2 mM to about 8 mM and an acetate concentration of about 37 mM to about 47 mM.

[0345] 17. The formulation of statement 16, wherein the sodium acetate buffer has a sodium concentration of about 5 mM and an acetate concentration of about 43 mM.

[0346] 18. An aqueous pharmaceutical formulation, the formulation comprising an anti- CD20 antibody in a concentration of 110 mg / ml, sucrose in a concentration of 180 mM, methionine in a concentration of 30 mM, polysorbate 20 in a concentration of 0.06% (w / v), hyaluronidase in a concentration of 2000 U / ml, polyvinylpyrrolidone (PVP) KI 2 in a concentration of about 7.5% (w / v), and an acetate buffer, and wherein the formulation has a pH of 5.0 to about 5.6, wherein the anti-CD20 antibody comprises a VH domain comprising the amino acid sequence set forth in SEQ ID NO: 8, a VL domain comprising the amino acid sequence set forth in SEQ ID NO: 7, and a human IgGl constant region, wherein the formulation is suitable for subcutaneous administration.

[0347] 19. The formulation of statement 18, wherein the formulation has a sodium acetate buffer, and wherein the sodium acetate buffer has a sodium concentration of 5 mM and an acetate concentration of about 43 mM.

[0348] 20. The formulation of any one of statements 1-19, wherein the weight average molecular weight of PVP KI 2 is 2000 Dalton to 7000 Dalton, optionally wherein the molecular weight is assessed by size exclusion chromatography (SEC) using light scattering detection, further optionally wherein the weight average molecular weight of PVP KI 2 is 2000 Dalton to 3000 Dalton.

[0349] 21. The formulation of any one of statements 1-20, wherein the anti-CD20 antibody comprises a light chain comprising the amino acid sequence of SEQ ID NO: 9 and a heavy chain comprising the amino acid sequence of SEQ ID NO: 10 or 11.

[0350] 22. The formulation of any one of statements 1-21, wherein the anti-CD20 antibody is ocrelizumab.64MF-363724269Attorney Docket No. 146392068840

[0351] 23. The formulation of any one of statements 1-22, wherein the hyaluronidase is recombinant human hyaluronidase.

[0352] 24. The formulation of any one of statements 1-23, wherein: a) the antibody in the formulation is stable at 2-8°C for at least 6 months, at least 12 months, at least 18 months or at least 24 months, b) the antibody and hyaluronidase in the formulation are stable after continuous agitation for 24 hours at room temperature, and / or c) there is i) less than 1% of aggregation of the antibody in the formulation as assessed by sum of HMW forms by SE-UHPLC after 25 days at 40°C and / or ii) less than 6% of degradation of the antibody in the formulation as measured by sum of pre-peaks by CE- SDS after 25 days at 40°C.

[0353] 25. The formulation of statement 24, wherein the hyaluronidase in the formulation is stable when it retains at least about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of its biological activity.

[0354] 26. The formulation of statement 24 or statement 25, wherein the antibody in the formulation is stable when it retains at least about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of its biological activity.

[0355] 27. The formulation of statement 26, wherein the biological activity is measured by antibody binding to CD20.

[0356] 28. The formulation of any one of statements 1-27, which is sterile.

[0357] 29. A formulation of any one of statements 1-28 for use in a method of treating multiple sclerosis in a patient comprising subcutaneously administering the formulation.

[0358] 30. The formulation for use according to statement 29, wherein the patient has not been subjected to a prior intravenous administration of the anti-CD20 antibody at a dose of about 600 mg.

[0359] 31. The formulation for use according to statement 29 or 30, wherein the patient has not been subjected to a prior intravenous administration of the anti-CD20 antibody.

[0360] 32. The formulation for use according to any one of statement 29-31, wherein the patient has not been subjected to a prior subcutaneous administration of the anti-CD20 antibody at a dose lower than about 900 mg.65MF-363724269Attorney Docket No. 146392068840

[0361] 33. The formulation for use according to statement 29, wherein the patient has received a prior treatment with an anti-CD20 antibody, optionally wherein the prior treatment comprises an intravenous administration of the anti-CD20 antibody, optionally wherein the prior treatment comprises the intravenous administration of the anti-CD20 antibody at a dose of about 300 mg or about 600 mg.

[0362] 34. The formulation for use according to any one of statements 29-33, wherein the method comprises subcutaneously administering more than one dose of the anti-CD20 antibody.

[0363] 35. The formulation for use according to of statement 34, wherein the method comprises administering at least two, three or four doses of the anti-CD20 antibody.

[0364] 36. The formulation for use according to any one of statements 29-35, wherein the anti-CD20 antibody is administered at a frequency of no more than once every 24 weeks or about once every 24 weeks.

[0365] 37. The formulation for use according to any one of statements 29-36, wherein the anti-CD20 antibody is administered at a frequency of no more than once every 6 months or about once every 6 months.

[0366] 38. The formulation for use according to any one of statements 29-37, wherein anti-CD20 antibody is the only medicament administered to the patient to treat multiple sclerosis.

[0367] 39. The formulation for use according to any one of statements 29-38, wherein the patient is subject to a) an oral dexamethasone or an equivalent corticosteroid and b) an oral antihistamine administration (e.g., desloratadine) shortly prior to the subcutaneous administration of the anti-CD20 antibody, optionally wherein the oral dexamethasone or the equivalent corticosteroid and antihistamine are administered at least or within 30 minutes, 20 minutes, or 15 minutes prior to the subcutaneous administration, further optionally wherein the patient is subject to 20 mg dexamethasone and 5 mg desloratadine.

[0368] 40. The formulation for use according to any one of statements 29-39, wherein the multiple sclerosis is relapsing multiple sclerosis (RMS).

[0369] 41. The formulation for use according to any one of statements 29-40, wherein the patient has a clinical isolated syndrome (CIS), relapsing-remitting multiple sclerosis (RRMS) or active secondary progressive multiple sclerosis (SPMS).66MF-363724269Attorney Docket No. 146392068840

[0370] 42. The formulation for use according to any one of statements 29-41, wherein the multiple sclerosis is a primary progressive multiple sclerosis (PPMS).

[0371] 43. The formulation for use according to any one of statements 29-42, wherein the patient has an Expanded Disability Status Scale (EDSS) score of 0-6.5.

[0372] 44. The formulation for use according to of any one of statements 29-43, wherein the subcutaneous administration takes no more than about 10 minutes.

[0373] 45. The formulation for use according to any one of statements 29-44, wherein said anti-CD20 antibody in the formulation is about 110 mg / ml.

[0374] 46. The formulation for use according to any one of statements 29-45, for use in a method of treating multiple sclerosis with reduced inflammation at the subcutaneous injection site, wherein inflammation is reduced compared with administration of the corresponding formulation in which the PVP K-12 is replaced with PVP K-17.

[0375] 47. PVP K-12 for use in a method of reducing inflammation at the injection site of a subcutaneously administered anti-CD20 antibody, wherein the PVP K-12 is formulated with the anti-CD20 antibody for subcutaneous administration, wherein optionally the PVP K- 12 is formulated with the anti-CD20 antibody in a formulation according to any one of statements 29-35.

[0376] 48. The formulation for use according to any one of statements 29-45, for use in a method of treating multiple sclerosis with increased bioavailability of the anti-CD20 antibody, wherein bioavailability is increased compared to administration of a pharmaceutical formulation containing the anti-CD20 antibody and PVP KI 2 at a concentration of 5% and / or 10%.

[0377] 49. PVP-K12 for use in a method of increasing bioavailability of a subcutaneously administered anti-CD20 antibody, wherein the PVP K-12 is formulated with the anti-CD20 antibody for subcutaneous administration in a formulation at 7.5 % PVP K-12, wherein optionally the PVP K-12 is formulated with the anti-CD20 antibody in a formulation according to any one of statements 29-45.

[0378] 50. The formulation for use according to any one of statements 29-49, wherein the anti-CD20 antibody comprises a light chain comprising the amino acid sequence of SEQ ID NO: 9 and a heavy chain comprising the amino acid sequence of SEQ ID NO: 10 or 11.67MF-363724269Attorney Docket No. 146392068840

[0379] 51. The formulation for use according to any one of statements 29-50, wherein the anti-CD20 antibody is ocrelizumab.EXAMPLES

[0380] The application will be more fully understood by reference to the following examples. They should not, however, be construed as limiting the scope of the application. It is understood that the examples and embodiments described herein are for illustrative purposes only and that various modifications or changes in light thereof will be suggested to persons skilled in the art and are to be included within the spirit and purview of this application and scope of the appended claims.Example 1: Developing a high-concentration CD20 antibody formulation for subcutaneous (SC) administration

[0381] There are multiple challenges in developing a high concentration antibody formulation for subcutaneous administration in a human patient. Such formulation needs to achieve at least a) desirable solubility not only in storage solution but also in subcutaneous space after it is administered, b) minimum toxicity and c) desirable bioavailability. For ocrelizumab specifically, previous subcutaneous tolerability studies conducted by the applicant with formulations having ocrelizumab concentrations above 50 mg / mL resulted in significant inflammation at the injection site. Further in vitro investigation into the potential cause for the injection site reaction revealed that ocrelizumab has a protein concentration dependent insolubility in high salt (and to a lesser extent, neutral pH) environments.

[0382] As discussed below, the inventors successfully developed high concentration ocrelizumab (e.g., 110-120 mg / mL) formulations for SC administration that are stable, well tolerated and achieved desirable systemic exposures.

[0383] The components of an exemplary high concentration ocrelizumab formulation for subcutaneous delivery are described in Table 1.Table 1: Components of exemplary high-concentration SC ocrelizumab formulation68MF-363724269Attorney Docket No. 146392068840

[0384] As described in the present application, a concentration at 20 mM of a sodium acetate buffer refers to an acetate concentration at 20 mM in the sodium acetate buffer that was used to prepare the formulation. A sodium acetate buffer with a concentration at 20 mM can be made, for example, by mixing 16 mM sodium acetate and 4 mM glacial acetic acid. However, because of the high concentration of the antibody and due to the Gibbs-Donnan effect and volume excursion effects during ultrafiltration and diafiltration (UF / DF) process, the final acetate concentration is about 43 mM (in the range of 37 mM to 47 mM) and the final sodium concentration is about 5 mM (in the range of 2 mM to 8 mM) in the formulation. Although 2000 U / ml is used for the formulation, inventors note that a much lower concentration (e.g., 500 U / ml) will work similarly to e.g., enhance tissue permeability and reduces injection pain / pressure.Preparation of Formulations

[0385] All ocrelizumab used in the studies below was prepared by using protein in 20 mM or 30 mM sodium acetate and concentrating the material using Amicon® Ultra 15mL or equivalent to achieve a pH of 5.0-5.6. Ocrelizumab used in the GLP study was prepared by concentrating in 20 mM sodium acetate using a tangential flow filtration system which is to achieve a pH of 5.0-5.6 (e.g., 5.3). Specifically, anti-CD20 antibody was produced and purified in a 20mM sodium acetate buffer. The purification process includes affinity chromatography, multimodal anion-exchange chromatography and hydrophobic interaction chromatography. The antibody was then subject to a small virus retentive filtration, ultrafiltration and diafiltration (UF / DF). A conditioning buffer with all other ingredients except hyaluronidase was prepared and mixed with the antibody; hyaluronidase was then added to obtain the final formulation. Protein concentration was tested using a UV- spectrophotometer.

[0386] Other proteins used as controls were used from filled drug product vials manufactured by Roche (Rituxan Hycela®).69MF-363724269Attorney Docket No. 146392068840Example 2: High-concentration ocrelizumab formulations that are well tolerated after subcutaneous injectionAnimal model and scoring system

[0387] The rat local tolerance model was selected as an appropriate and validated species for assessing SC ocrelizumab formulations. Toxicity studies in cynomolgus monkeys, minipigs, and rats administered ocrelizumab not formulated for SC injection revealed similar injection site reactions based on histologic findings.

[0388] In the rat SC local tolerance models described for each study following, 0.25-0.5 mL of test formulation were administered subcutaneously to the lower or upper dorsal region of the animal. Prior to dosing and as needed thereafter, the area used for dosing was clipped of hair, and dose sites were marked and maintained as needed. Local tolerance evaluation was based on the Draize skin irritation scoring system (see Table 2), clinical observations, body weights, and macroscopic and microscopic anatomic pathology. Clinical observations were performed daily for the first 3-4 days or as indicated for each study and injection sites were examined for redness or swelling and were recorded and scored based on an established scoring scale (Draize, 1959). Skin samples at the site of injection were collected and bisected (without separating the two halves) at necropsy for routine histology. Skin samples collected during necropsy were placed into 10% neutral buffered formalin (NBF) for fixation. The skin samples were trimmed to produce histological sections. A skin sample from each animal in all dose groups was embedded in paraffin, processed in the routine manner, sectioned, stained with hematoxylin and eosin, and examined microscopically.Table 2: Scoring scale for injection site inflammation (based on scoring scale of Draize 1959)70MF-363724269Attorney Docket No. 146392068840Study A.Materials and Methods

[0389] Three rats per group were subcutaneously administered a single dose of test article in a fixed dose volume of 0.5 mL. All test articles in this study were formulated in 2000 U / ml rHuPH20 (“PH20”), Sodium Acetate 30 mM, Sucrose 240 mM, Methionine 30 mM, polysorbate 20 0.06%, pH 5.3.

[0390] Cohort 1 tested formulations containing no PVP, high molecular weight PVP (K17; 10%) and various levels of low molecular weight PVP (K12; 1%, 2.5%, 10%) with ocrelizumab at a concentration of 135 mg / mL.

[0391] Cohort 2 tested formulations containing various levels of K12 PVP (5%, 10%) and K17 PVP (10%) with ocrelizumab at a lower concentration of 120 mg / mL.

[0392] The design of the study is described below in Table 3.Table 3: Design of rat toxicity studyResults and Conclusions

[0393] The score system is shown in Table 6. Cohort 1 and cohort 2 results are shown inTable 4 and Table 5, respectively.

[0394] Specifically, in cohort 1, overall, minimal Draize scoring (minimal to no redness and swelling) was observed on skin irritation. However, upon histopathological assessment, marked inflammation, characterized by mixed cell infiltrate with degenerate71MF-363724269Attorney Docket No. 146392068840 neutrophils / edema and lymphocytes accompanied by small numbers of macrophages with intracellular hyaline material (presumptive ocrelizumab), was observed in control groups by the end of study on Day 4. In the formulations prepared with K12, inflammation increased with decreasing amounts of PVP, with mild-moderate inflammation with 10% K12, minimal- marked inflammation with 2.5% K12, and severe inflammation with 1% K12. These results indicate the unsuitability of these 135 mg / ml formulations to be used for subcutaneous administration in human.

[0395] In cohort 2, overall, minimal Draize scoring (minimal to no redness and swelling) was observed on skin irritation. Upon histopathological assessment, minimal-mild inflammation was observed in all KI 2 groups and mild-moderate inflammation in the K17 group by the end of study on Day 4. Specifically, in this cohort of 120 mg / ml ocrelizumab formulations those with KI 2 were better tolerated than formulations with K17.Table 4: Cohort 1 results72MF-363724269Attorney Docket No. 146392068840Table 5: Cohort 2 results73MF-363724269Attorney Docket No. 146392068840Table 6: Injection Site Severity ScoresStudy B.

[0396] Study B was a GLP rat subcutaneous injection tolerability study comparing ocrelizumab in the PVP-containing high concentration formulation against a control of Rituxan Hycela®, the marketed product containing rituximab for subcutaneous injection. Ocrelizumab and vehicle are formulated in 20 mM sodium acetate, 240 mM sucrose, 30 mM methionine, 0.06% polysorbate 20, 2000 U / ml rHuPH20, and 7.5% PVP K12. Rituxan Hycela® is a high concentration SC formulation of an anti-CD20 antibody (rituximab; 120 mg / ml) approved for treatment of autoimmune disorders as well as for some types of cancer. Each mL of Rituxan Hycela® solution contains rituximab (120 mg), hyaluronidase human (2,000 Units), L-histidine (0.53 mg), L-histidine hydrochloride monohydrate (3.47 mg), L- methionine (1.49 mg), polysorbate 80 (0.6 mg), a,a-trehalose dehydrate (79.45 mg), and water for injection.

[0397] The local tolerability of high concentration ocrelizumab formulation was assessed with a subcutaneous (SC) administration of 0.5 mL ocrelizumab formulation to male Sprague Dawley rats as a single dose or repeat doses (Q2W x2) for 4 weeks. Vehicle and Rituxan Hycela® were used as controls. Rituxan Hycela® was added as a high protein concentration control as this formulation was previously demonstrated to be well tolerated in both nonclinical and clinical studies. The reversibility, persistence, or delayed occurrence of any effects were evaluated after a 3- and 14-day observational period or after a 14-day74MF-363724269Attorney Docket No. 146392068840 observational period. Additionally, a toxicokinetic profile was obtained from the repeat-dosed rats (Group 3).Materials and Methods

[0398] Specifically, the study design is described in FIG. 1. There were three groups of animals dosed in this study. In the single dose group (n= 12), animals received a single 0.5 mL SC injection of both the vehicle control and ocrelizumab formulation at two separate injections sites on Study Day 1, and animals were necropsied on Days 4 and 15. In the repeat dose group (n=6), animals received 0.5 mL SC injection of both vehicle control and ocrelizumab formulation at two separate injections sites on Study Days 1 and 15, and animals were necropsied on Day 29. Additionally, Rituxan Hycela® (n=6) was included in the study as a comparator group.

[0399] Assessment of tolerability was based on mortality, clinical observations, body weights, food consumption, dermal observations, and anatomic pathology of the dose sites, as described in previous studies. Blood samples were collected for toxicokinetic evaluations from Group 3.Results and Conclusions

[0400] All animals survived to their scheduled necropsy. No formulation-related clinical, dermal, or macroscopic observations or alterations in body weight or food consumption were noted in any groups.

[0401] Minimal-slight inflammatory findings were noted microscopically at both dose sites of all animals. The incidence and severity of findings were generally similar between animals and injection sites whether administered Rituxan Hycela® or ocrelizumab formulation on Study Day 1 only or on Study Days 4, 15, and 29. See FIG. 2.

[0402] Exposure to high concentration ocrelizumab, as assessed by mean Cmax and AUC values, was generally similar on TK Days 0 and 14 (corresponding to Study Day 1 and 15), with mild accumulation after multiple doses. Ocrelizumab formulation’s mean Tmax values were 2.67 days on TK Day 0 and 1.33 days on TK Day 14. The concentration of the antibody after the administration remains constant for at least 10 days after it reaches its maximum in the serum. See FIG. 3.

[0403] Systemic ocrelizumab exposure was confirmed and generally similar upon single and repeat SC administration at the 60 mg dose level over a 4- week period. Exposure to the ocrelizumab formulation following the first and second SC dose administrations was assessed75MF-363724269Attorney Docket No. 146392068840 by the mean maximum concentration (Cmax) and the area under the concentration-time curve within the 14-day dosing interval (AUC). The mean Cmax after the first and second administrations were 544 pg / mL and 707 pg / mL, respectively, while the mean AUC values were 5940 day* pg / mL and 6240 day* pg / mL, respectively. The mean time to peak drug concentration (Tmax) following the first and second doses was 2.67 days and 1.33 days, respectively (FIG. 4).

[0404] In conclusion, subcutaneous administration of 0.5 mL of Ocrelizumab formulation [120 mg / mL high-concentration ocrelizumab (2000 U / ml rHuPH20 and 7.5% K12 PVP)] was well tolerated. The findings from this group are comparable to the high- concentration vehicle control and Rituxan Hycela® comparator groups. This dose level of ocrelizumab high concentration formulation corresponded to mean maximum observed concentration (Cmax) and area under the concentration-time curve from 0 to 14 days post second dose (AUC 14-28) values of 707 g / mL and 6240 day*pg / mL, respectively, in male Sprague Dawley rats on Study Day 15.Example 3: The high concentration ocrelizumab formulation achieves a desirable biodistribution

[0405] This example describes experiments investigating the biodistributions, bioavailability, and systemic exposure achieved by various ocrelizumab formulations after subcutaneous administration.MethodsAnimal Studies

[0406] Female Sprague Dawley rats, ranging in 200-220 g, were randomly assigned into groups of 3. At 24h and Ih prior to dose administration, rats received 0.1 mL intraperitoneal injections of 30 mg / kg sodium iodide in saline solution to prevent thyroid sequestration of radioactive iodine. All animals received a single 0.5 mL flat dose of 120 mg / mL ocrelizumab with various formulations as outlined in Table 7. Following intravenous (IV) or interscapular subcutaneous (SC) injections, rats were anesthetized with isofluorane and blood sampled via jugular vein at 2 hours (h), 3 h, 6 h, 1 day (d), 2 d, 3 d, 7 d, 14 d and 21 d. Whole blood was processed for plasma using lithium heparin microtainers. 50 pL of whole blood, 50 pL of plasma and 1-5 pL aliquots of reference doses were counted on a Wizard2 gamma counter76MF-363724269Attorney Docket No. 146392068840(Revvity) to determine the percent of injected dose per mL (%ID / mL). Radio- specific activity was then used to convert %ID / mL values into pg antibody per mL.Table 7. Group Dose FormulationDose formulation, antibody concentration, pH and osmolality are summarized for each group. All groups received a single 0.5 mL of dose material as indicated by the dose route. Abbreviations: IV is intravenous, SC is subcutaneous in the interscapular area.Radiochemistry

[0407] Ocrelizumab formulated in 30 mM sodium acetate pH 5.3 was first buffer exchanged into 10 mM phosphate buffered 150 mM NaCl (PBS) using NAP5 desalting columns (GE Healthcare). 1 mCi of sodium [125I]iodide was then oxidized in lodogen tubes (Perkin Elmer) for 5 minutes before being transferred to a separate tube containing 75 pg of antibody. The indirect iodination was conducted for 1 min before unreacted iodine was removed using NAP5 columns. Following buffer exchange into 30 mM sodium acetate pH 5.4 and concentration with a 10 kDa molecular weight cutoff spin column (Millipore), radio purity was confirmed by using SEC-HPLC by resolving antibodies on an Acquity UPLC Protein BEH SEC 1.7pm 200A column (Waters) with isocratic PBS for 10 min.SPECT-CT Imaging

[0408] Rats were anesthetized under sevofluorane for SPECT-CT (MILabs) imaging at 2h, Id, 2d and 3d following dose administration. 20 min imaging sessions were acquired with the77MF-363724269Attorney Docket No. 146392068840 high-energy general-purpose vector collimator with 3.6 mm pinholes. Each image was reconstructed using a SROSEM reconstruction method (MILabs, version 11) at 0.8 mm with 4 subsets and 4 iterations within the 15-45 keV window for 1251. Images were then corrected for attenuation and co-registered to the CT images at 200 pm voxel resolution. Resulting images were rendered for viewing and analyzed using VivoQuant™ software (InVicro) using 3D voxel annotation. Briefly, volumes of interest were segmented to include the torso above the liver and excluding the chest cavity as demarked by the connecting edges of the lungs as identified by CT. The subcutaneous bolus was then segmented from the torso by thresholding for signal greater than or equal to 1 %ID / g. All data were plotted in PRISM (GraphPad) software. Data are represented as mean values + / - SD.Pharmacokinetic Analysis

[0409] Samples were subjected to gamma counting alongside dose standards to determine the %ID / mL of each sample. Then, using the radiospecific activity, values were converted to pg / mL of ocrelizumab. Serum radio-concentration-time profiles were used to estimate the PK parameters in rat using non-compartmental analysis (NCA) (Phoenix™ WinNonlin®, Version 6.4; Pharsight Corporation; Mountain View, CA, USA). Each animal was analyzed separately, and results for each dose group were summarized as mean + / - standard deviation (SD). Antibody half-life, highest observed concentration (Cmax), time of observed Cmax (Tmax), area under the curve extrapolated to infinity (AUC0-co) and clearance (CL_F) were calculated. AUC was determined using the log-linear trapezoidal rule. Bioavailability was calculated by dividing the AUC0-co for the subcutaneous dosed groups by the intravenous dosed group. Nominal sample collection times and nominal dose solution concentrations were used in PK analysis. No animals were excluded from the statistical analysis.ResultsGeneration of imaging agents

[0410] Ocrelizumab was radiolabeled with 1251 randomly through tyrosine residues using the indirect iodination method to reduce antibody oxidative damage. Incorporation of 1251 at the given reaction conditions was 46%, which is consistent with this labeling method. The resulting in a radiospecific activity was 6.4 uCi / pg of antibody. The molar labeling ratio was 0.45, indicating a low probability of a single antibody having more than one radiolabel (Table 8).78MF-363724269Attorney Docket No. 146392068840Table 8. Radiolabeling ResultsRadiolabeling product information is summarized in the table. Radiolabeling efficiency, radioactivity concentration, antibody concentration, radiospecific activity and molar incorporation of activity are listed.

[0411] Even though a fraction of antibodies were radiolabeled, this is common practice for measuring radio-PK and distribution. Greater than 99% radiopurity was confirmed using size exclusion chromatography, fitted with both absorbance and radio-detection inline serial sensors. Free radio iodine has a later retention time of 7.8 min, and was absent in radiolabeled stock preparations and dose formulations. Radiolabeled ocrelizumab showed a retention time of 4.3 min, while the A280 protein peak for both radiolabeled and unlabeled ocrelizumab was observed at 3.9 min. The aligned retention time, along with the absence of shoulder peaks in either radio or protein signal indicated that the radiolabeling method did not induce dramatic protein degradation or aggregation. Radiolabeled ocrelizumab was shown to exhibit a consistent, but slightly left-shifted retention time of 4.1 min when added to dose formulation of 120 mg / mL ocrelizumab for IV dosing (30 mM sodium acetate pH 5.3, 240 mM Sucrose, 30 mM Methionine, 0.06% w / v polysorbate 20) or SC dosing (same components as IV with additional 2000 units / mL Hyaluronidase and PVP). Slight left temporal shifts are to be expected in SEC-HPLC traces when high concentrations of protein are co-injected (FIG. 5).SPECT-CT Imaging

[0412] Distribution of radiolabeled ocrelizumab was imaged with SPECT-CT static imaging at 2 h, 1 d, 2 d and 3 d post single IV or SC dosing. At the earliest time point of 2 h, the IV dose group showed the highest systemic exposure as seen by signal in the heart, descending aorta and general vascularized tissue. The SC dose group without PVP formulation exhibited a smaller bolus with higher antibody concentration compared to the other three groups formulated with PVP. 1 d following dose administration, the IV dosed group showed generalized systemic distribution, with faint splenic uptake. In contrast, all of the SC dosed groups showed an expanded SC bolus, with faint distribution highlighting more pronounced splenic uptake. On the second day, slight thyroid uptake of free 1251 is observed for all dose groups, which is expected without continued thyroid block with nonradioactive sodium iodide. All PVP formulated doses showed a significant drop in signal associated with the previous SC bolus site. This was in contrast to the SC formulation group lacking PVP, which79MF-363724269Attorney Docket No. 146392068840 showed a consistent retention of ocrelizumab in the SC dosing site at a high concentration. This SC bolus retention for the non PVP formulation group continued to the final imaging day (3 d) (FIG. 6).

[0413] Dose material in the subcutaneous space was measured by generating a large volume of interest and further segmenting the activity over a threshold of 1 %ID / g. The activity measured as subcutaneous over 1 %ID / g was plotted as the percent of injected dose (FIG. 7). SC bolus volumes of interest (VOI) were segmented from a larger VOI of the back of the animal ranging from the base of the neck to the bottom of the rib cage from images shown in FIG. 7. The SC bolus was then segmented by thresholding the signal activity to be greater than or equal to 1 %ID / g. The data was then converted to %ID based on the volume of the VOI, assuming the tissue density to be that of water.

[0414] Time points analyzed are listed in Table 9. Animals were bled and imaged according to the time points indicated.Table 9. Animal study timepoints

[0415] Ocrelizumab dosed without PVP formulation showed the highest concentration in the SC space throughout the 3-day imaging portion of the study. Formulation with 5% PVP improved the SC clearance, but was still at a detectable range on the third day. This is in contrast to formulation with both 7.5% and 10% PVP reduced the signal to undetectable ranges by the third day. We observed that the formulation with 7.5% PVP showed the fastest reduction in signal of the concentrations tested.Pharmacokinetics and Subcutaneous Bioavailability

[0416] Systemic exposure of ocrelizumab was evaluated as radioactivity measurement in plasma collected at 2 h, 3 h, 6 h, 1 d, 2 d, 3 d, 7 d, 14 d and 21 d post dose (FIG. 8).80MF-363724269Attorney Docket No. 146392068840

[0417] Non-compartmental analysis method was used to determine pharmacokinetic parameters based on plasma concentration-time profiles (Table 10). All PVP formulated groups had a significantly higher Cmax compared to the non-PVP SC dosed group, with the 7.5% PVP formulated group showing the highest Cmax of 1759 ug / mL. The 7.5% PVP formulation group also had the highest AUCO-co (15,179 d*ug / mL), lowest CL_F (4 mL / d / kg) and highest bioavailability (73%) out of all of the SC formulation groups.Table 10. Pharmacokinetic parameters of ocrelizumab in rats based on Noncompartmental Analysis of Systemic Exposure.NCA values are summarized for all dose groups. Data represent averages (n=3) for each group ± standard deviation. Bioavailability was calculated by dividing AUCo-o of the SC group to the average AUCo-o of the IV dosed group.

[0418] As above, Ocrelizumab formulations with a 7.5% PVP achieved an unexpectedly advantageous high Cmax, AUC0-co , and SC bioavailability.Example 4: 7.5% PVP K12 enhances solubility of anti-CD20 antibody in high concentration formulation in PBS pH 7.2-7.4 (which corresponds to the pH of subcutaneous space)

[0419] The solubility of ocrelizumab was measured by assessing the turbidity of the sample. The turbidity test is a measure of light scattering of the sample using UV- Visible spectrophotometer (or automated Microplate Reader). The formulated samples were monitored at wavelengths 340, 345, 350, 355, and 360 nm and the average scattering at these wavelengths was reported as the turbidity of the sample (FIG. 9). Control (CTRL) corresponds to anti-CD20 antibody at 110 mg / mL with 0% PVP. A turbidity reading of 0.5 OD or above is considered significant precipitation of ocrelizumab. As shown, 7.5% PVP KI 2 allows minimum turbidity for a formulation with an antibody concentration of 110 mg / mL to 140 mg / mL at a pH of 7.2 to 7.4.81MF-363724269Attorney Docket No. 146392068840Example 5: Povidone K12 reduces surface tension of the solution and can act as a surfactant to improve agitation stability of anti-CD20 antibody at high concentrationSummary

[0420] The purpose of this example is to investigate if povidone K12 can act as a surfactant in addition to its main function as a solubilizer. The results demonstrate that povidone K12 can act as a surfactant and prevent degradation of ocrelizumab high concentration at the airliquid interface during agitation stress. The agitation stability was assessed in the ocrelizumab subcutaneous (SC) high concentration (HC) drug product composed of 110 mg / mL ocrelizumab and 2000 U / ml rHuPH20 formulated in 20 mM sodium acetate (As discussed above, due to the Gibbs-Donnan effect and volume excursion effects, the final sodium concentration is about 5 mM (in the range of 2 mM to 8 mM), and the final acetate concentration is about 43 mM (in the range of 37 mM to 47 mM) in the formulation.), 240 mM sucrose, 30 mM methionine at pH 5.3 with 0 or 50 mg / mL povidone K12 and / or with 0 to 0.4 mg / mL of polysorbate 20. The results showed that 50 mg / mL povidone K12 is effective at preventing drug product degradation by protein denaturation at the air-liquid or liquid- surface interface.Materials and Methods

[0421] Five formulations of ocrelizumab HC, as shown in Table 11, were assessed for the agitation stress study. A concentration of 50 mg / mL povidone KI 2 is used in this study because it is the lowest end of the acceptable specification.Table 11: Overview of ocrelizumab SC high concentration formulations for the agitation stability study.aAs discussed above, due to the Gibbs-Donnan effect and volume excursion effects, the final sodium concentration is about 5 mM (in the range of 2 mM to 8 mM), and the final acetate concentration is about 43 mM (in the range of 37 mM to 47 mM) in the formulation.82MF-363724269Attorney Docket No. 146392068840

[0422] Three sets of samples were prepared for the agitation study. Each set contained 5 formulations and was subjected to the following:1. A fill volume of 4 mL of each formulation in autoclaved 6 mL Type 1 glass vials; a total of three vials per formulation.2. One set of the samples was placed at 2-8°C as TO.3. One set of the sample was placed horizontally on the arm shaker set at 100 rpm at ambient temperature and agitated continuously for 24 hours.4. One set of the sample was placed next to the arm shaker for 24 hours without agitation at ambient temperature as the control vials.

[0423] The following product quality attributes were assessed: visible particles by visual inspection (VP), CAC, pH, protein concentration, SE-UHPLC, and enzyme (rHuPH20) activity.

[0424] In addition, the surface tension of the ocrelizumab HC samples containing different combinations of povidone K12 and / or polysorbate 20, as shown in Table 12, was measured. The surface tension measurements were made using the Biolin Scientific Attension Theta Pendant Drop tensiometer. In this technique, the shape of a pendant solution drop image is fitted to the surface tension as a function of time. The measurements were taken up to 120 seconds from the creation of a fresh drop.Table 12: Overview of ocrelizumab SC high concentration formulations for the surface tension test83MF-363724269Attorney Docket No. 146392068840Results / Discussion

[0425] The following was observed:1. As expected, in the absence of a surfactant such as polysorbate 20 or povidone K12 (positive control), significant increases in visible particles, solution opalescence, HMW forms, and complete loss of rHuPH20 activity were observed after 24 hours of agitation compared to the control (TO and 24h without agitation) (FIGs. 10A-10B).2. In the sample with 50 mg / mL povidone K12 (without a surfactant such as polysorbate 20), it is surprisingly found that the solution opalescence was significantly improved compared to the positive control. In addition, no increases in HMWF were observed. The rHuPH20 activity significantly improved from no activity detected in the positive control to only a slight decrease (11%) in rHuPH20 activity in the sample formulated with 50 mg / mL povidone (without polysorbate 20).3. No product quality impact was observed for the samples formulated with 50 mg / mL povidone K12 and with polysorbate 20 levels of 0.3 and 0.4 mg / mL after 24h of agitation.

[0426] The surface tension of the formulation buffers with and / or without povidone KI 2 and / or polysorbate 20 as well as the drug product at 110 mg / mL (HC) and 40 mg / mL (low concentration) was assessed and shown in FIG. 11. The following was observed:1. The presence of 0.6 mg / mL polysorbate 20 in the formulation buffer or drug product reduces the surface tension in comparison to the formulation without povidone and polysorbate 20 (positive control).2. The presence of 75 mg / mL povidone K12 in the formulation buffer (without polysorbate) reduces the surface tension in comparison to the formulation without povidone and polysorbate 20. However, 75 mg / mL povidone K12 is not sufficient to reduce the surface tension to the same level as the formulation buffer or DP containing 0.6 mg / mL polysorbate 20.

[0427] The data shows that povidone KI 2 can reduce the surface tension compared to the positive control (without povidone K12 and polysorbate 20) and function as a surfactant. The reduction in surface tension by povidone KI 2 and its function as a surfactant explains the84MF-363724269Attorney Docket No. 146392068840 improved solution opalescence and rHuPH20 activity as well as no increases in HMW forms (when povidone K12 is present in the formulation without polysorbate 20) after agitation for 24 hours.Conclusion

[0428] Povidone KI 2 can function as a surfactant to lower the surface tension and prevent product degradation at the air-liquid interface during agitation on its own (without polysorbate 20).Example 6: High concentration formulation provides excellent stability for the anti- CD20 antibody

[0429] This example assesses the stability of ocrelizumab subcutaneous (SC) high concentration (110 mg / mL) drug product (DP) and ocrelizumab SC low concentration (40 mg / mL) DP under the stressed condition of 40°C. This stress comparability study was conducted with ocrelizumab high concentration DP and low concentration stored in parallel. The degradation rates and profiles by SE-UHPLC, IE-HPLC, and non-reduced CE-SDS were compared between the two drug products. The results demonstrated excellent charge and fragmentation stability of the ocrelizumab high concentration formulation after storage at 40°C / 75% RH (relative humidity) for 28 days, as shown by IE-HPLC and non-reduced CE- SDS.Material and Methods

[0430] The ocrelizumab high concentration (110 mg / mL) and low concentration (40 mg / mL) DP with composition listed in Table 13 were used in these studies. Ocrelizumab high concentration and low concentration DP vials were placed side-by-side stability at 40°C / 75% RH, with samples pulled at 0, 7, 14, and 28 days.Table 13. Comparison of Ocrelizumab SC Low Concentration and High Concentration Formulations85MF-363724269Attorney Docket No. 146392068840NA = not applicableaSodium acetate trihydrate and glacial acetic acid are added to reach the target solution pH of 5.3.bDue to the Gibbs-Donnan effect and volume excursion effects, the final sodium concentration is about 5 mM (in the range of 2 mM to 8 mM), and the final acetate concentration is about 43 mM (in the range of 37 mM to 47 mM) in the formulation.

[0431] The following product quality attributes were assessed: Visual Inspection (VP), color, clarity / opalescence, pH, protein concentration, osmolality, IE-HPLC, SE-UHPLC, CE-SDS, potency, Polysorbate 20 content, povidone K12 content (only ocrelizumab high concentration), and subvisible particles by Light Obscuration.Results / Conclusion

[0432] Little to no changes were observed in the ocrelizumab high concentration (110 mg / mL) or low concentration (40 mg / mL) DP with respect to VP, color, clarity / opalescence, pH, protein concentration, potency, polysorbate 20, povidone K12 content, and subvisible particles after storage at 40°C for 28 days.

[0433] The ocrelizumab high concentration (110 mg / mL) DP has excellent charge variant and fragmentation stability trends, as demonstrated by IE-HPLC and non-reduced CE-SDS, which suggests that povidone KI 2 provides charge variants and fragmentation stability of ocrelizumab (FIG. 12). The aggregation rate measured by SE-UHPLC is higher in the high concentration DP as shown by the increase in the % sum of HMWF (corresponding to decrease in % main peak) (FIG. 12). It is an expected stability trend for ocrelizumab high concentration DP, as its protein concentration is approximately 3 times higher than that of the low concentration DP.

[0434] Degradation trends and profiles were assessed between the low (40 mg / mL) and high concentration (110 mg / mL) DP’s. The results demonstrate excellent stability profiles for the high concentration DP after storage at 40°C / 75% RH for 28 days and suggests that povidone KI 2 provides stability of ocrelizumab high concentration.86MF-363724269Attorney Docket No. 146392068840Example 7: Formulation with sucrose at a concentration of 180 mM is more effective in preventing aggregation and degradation of anti-CD20 antibody

[0435] As shown in this Example, an anti-CD20 formulation with sucrose at a concentration of 180 mM is more effective in preventing aggregation and degradation as compared to a similar formulation with sucrose at a concentration of 240 mM.Summary

[0436] The objective of this study was to evaluate the stability behavior of ocrelizumab subcutaneous (SC) high concentration (HC) drug product (DP) formulated with 240 mM sucrose and 180 mM sucrose under the stressed condition of 40°C / 75% relative humidity (RH) in a side-by-side stress stability study. After 28 days of storage at 40°C / 75% RH, results demonstrated 180 mM sucrose is more effective at preventing aggregation and degradation of ocrelizumab than samples formulated with 240 mM sucrose.Materials and Methods

[0437] The following ocrelizumab SC high concentration drug product formulations were used for the stress stability study:1. Drug product composed of 110 mg / mL ocrelizumab and 2000 U / ml of rHuPH20 (hyaluronidase) formulated with 20 mM sodium acetate, 240 mM sucrose, 30 mM methionine, 75 mg / mL povidone K12, 0.6 mg / mL polysorbate 20 at pH 5.3.2. Development drug product composed of 110 mg / mL ocrelizumab and 2000 U / ml of rHuPH20 (hyaluronidase) formulated with 20 mM sodium acetate, 180 mM sucrose, 30 mM methionine, 75 mg / mL povidone K12, 0.6 mg / mL polysorbate 20 at pH 5.3.3. GMP drug product composed of 110 mg / mL ocrelizumab and 2000 U / ml of rHuPH20 (hyaluronidase) formulated with 20 mM sodium acetate, 180 mM sucrose, 30 mM methionine, 75 mg / mL povidone K12, 0.6 mg / mL polysorbate 20 at pH 5.3.

[0438] As discussed above, due to the Gibbs-Donnan effect and volume excursion effects, the final sodium concentration is about 5 mM (in the range of 2 mM to 8 mM), and the final acetate concentration is about 43 mM (in the range of 37 mM to 47 mM) in above three formulations.

[0439] The ocrelizumab SC HC DP vials were placed side-by-side for stability testing at 40°C / 75% RH, with samples pulled at 0, 7, 14, and 28 days. The following product quality attributes were assessed: Visual Inspection (VP), color, clarity / opalescence, pH, protein87MF-363724269Attorney Docket No. 146392068840 concentration, IE-HPLC, SE-UHPLC, CE-SDS, potency, and subvisible particles by Light Obscuration.Results / Conclusion

[0440] The results demonstrate that the samples formulated with 180 mM sucrose show similar or lower level of degradation rates relative to samples formulated with 240 mM sucrose (refer to FIGs. 13A-13B; FIGs. 14A-14B, FIGs. 15A-15B). Lower degradation rates were observed in the percent sum of pre-peaks (LMW forms) and main peak by NR CE-SDS for the samples formulated with 180 mM sucrose (FIGs. 15A-15B). Little to no changes were observed in samples formulated with 180 mM or 240 mM sucrose with respect to visible particles, sub-visible particles, color, clarity and opalescence, pH, and protein concentration.Example 8: Povidone K12 is an excellent cryoprotectant and stabilizes anti-CD20 monoclonal antibody formulations during freeze / thaw and frozen storage

[0441] This example summarizes the study to investigate if povidone K12 can act as a cryoprotectant in the ocrelizumab high concentration (HC) formulation. The results demonstrated that in the presence of 50 mg / mL povidone (and no sucrose) in the formulation, no change in the % HMW forms by size exclusion chromatography (SE-UHPLC) after frozen storage at -8°C for 30 days and up to 5X freeze and thaw (F / T) cycles at ambient temperature and -20°C. The results suggest that povidone K12 is an excellent cryoprotectant and stabilizes ocrelizumab high concentration during frozen storage and from freeze / thaw stress. Introduction

[0442] Sucrose has historically been used as a cryoprotectant to stabilize mAbs during storage by preventing or reducing the formation of water crystals during freezing. In these studies, povidone K12 (also known as polyvinylpyrrolidone, or PvP) was investigated to determine if it can also act as a cryoprotectant in addition to its main function as a solubilizer. This study evaluated the % HMW forms by SE-UHPLC of ocrelizumab HC DS formulated with and without sucrose or / and povidone K12 after 5X F / T cycles at -20°C and ambient temperature as well as frozen storage at -8 °C (-8 °C was selected to accelerate frozen storage instability). The results showed both sucrose and povidone K12 can act as a cryoprotectant and protect ocrelizumab during frozen storage at -8 °C and up to 5X freeze / thaw (F / T) at -20°C and ambient temperature.88MF-363724269Attorney Docket No. 146392068840Materials and Methods

[0443] Three formulations of ocrelizumab HC, where the sucrose and povidone K12 levels were varied, were investigated. Ocrelizumab HC formulation composed of 110 mg / mL ocrelizumab in 20 mM sodium acetate (due to the Gibbs-Donnan effect and the volume exclusion effects, the sodium concentration is 5 mM, and the acetate concentration is 43 mM nominally in the final drug substance and drug product) at pH 5.3, 30 mM methionine, 0.6 mg / mL polysorbate 20 with either no sucrose and no povidone K12, no sucrose and with 50 mg / mL povidone K12, or with 240 mM sucrose and no povidone K12 were studied in the -8°C, -20°C and 5X F / T stability studies. A concentration of 50 mg / mL povidone K12 is used in this study because it is the lowest end of the acceptable specification.

[0444] Three sets of samples were prepared for the -8°C, -20°C and F / T stability studies.

[0445] Each set contained 3 formulations and was subjected to the following:1. A fill volume of 2 mL of each formulation into autoclaved 2 mL Type I glass vial; a total of three vials per formulation.2. Three sets of samples were stored at -40°C for a minimum of 2 hours to ensure the solution is completely frozen prior to -8°C and FT study.3. One set of samples was placed at -8°C (short-term frozen storage) with samples pulled at 0, 14, and 30 days.4. One set of samples was placed at -20°C (F / T study) with samples pulled at0, 3x and 5x F / T. Between each F / T cycle, samples were stored at -40°C for a minimum of 2 hours, followed by overnight storage at -20°C, and then thawed at room temperature for a minimum of 2 hours.

[0446] Frozen storage instability is expected to result in an increase in HMW forms; therefore, HMW forms were monitored by SE-UHPLC as it is the most stability-indicating and sensitive method.Results

[0447] The following was observed based on the ocrelizumab HC formulated with 50 mg / mL povidone K12 (no sucrose) or with 240 mM sucrose (no povidone K12, negative control):• No product quality impact (no increases in HMW forms) was observed after 5X F / T cycles as well as after 30 days of storage at -8°C.• No differences in degradation rates for SE-UHPLC in %main peak and %sum of HMW forms were observed after 5X F / T cycles (FIGs. 16A-16B) and after frozen storage at -8°C for 30 days (FIGs. 17A-17B).89MF-363724269Attorney Docket No. 146392068840• No new peaks in SE-UHPLC chromatographic profiles were observed after 5X F / T cycles (FIG. 16C) and after frozen storage at -8°C for 30 days (FIG. 17C).

[0448] The following was observed based on the ocrelizumab HC formulated without sucrose and without povidone (positive control) as expected:• A decrease of 1.7 in percent main peak and an increase of 1.7 in percent sum of HMW forms by SE-UHPLC after 5X F / T cycles.• A decrease of 0.8 in percent main peak and an increase of 0.8 in percent sum of HMW forms by SE-UHPLC after frozen storage at -8°C for 30 days.Conclusion

[0449] Povidone K12 at 50 mg / mL (in the absence of sucrose) substantially prevented any increases in the HMW forms. The results show that povidone KI 2 is an excellent cryoprotectant and is as effective as sucrose at preventing the formation of HMW forms during frozen storage of monoclonal antibodies.Example 9: Povidone KI 2 slows down polysorbate degradation by enzymatic hydrolysis and particle formation

[0450] The goal of this study is to evaluate the effect of the presence of polyvinylpyrrolidone (PVP, povidone K12) on free fatty acid (FFA) particle formation and polysorbate 20 (PS20) degradation.Introduction

[0451] Ocrelizumab high concentration is formulated in presence of PVP K12, a polymer that improves the solubility of the active pharmaceutical ingredient, ocrelizumab, in the subcutaneous space. However, the potential effects of PVP KI 2 on other components of the formulation are not well understood. The purpose of this study was to elucidate whether the presence of PVP KI 2 in the formulation could improve the solubility of free fatty acids resulting from polysorbate degradation.

[0452] PS20 is known to degrade through a hydrolytic mechanism into free fatty acids with limited solubility in aqueous solutions. Experiments were conducted similarly to Doshi, N., et al. (2021), but modified to compare a formulation with / without PVP (Doshi, N., et al. (2021) Pharm Res 38, 531-548). Two PS20 hydrolyzing enzymes were individually added to buffer containing PS20 with and without PVP. After the addition of PS20 hydrolyzing enzymes, visible particles, subvisible particles, and PS20 concentrations were measured at set time points over two days. PS20 concentrations were determined by UHPLC-ELSD.90MF-363724269Attorney Docket No. 146392068840

[0453] This data aims to provide insight into the effect of polyvinylpyrrolidone on FFA particle formation and polysorbate degradation.Materials and Methods

[0454] All experiments were conducted using complete formulation buffer composed of 20 mM sodium acetate, 30 mM methionine, 240 mM sucrose, 0.6 mg / mL polysorbate 20 at pH 5.3, with or without the presence of PVP. The following polysorbate-degrading enzymes were added:• Pseudomonas cepacia lipase (PCL) Item # 62309-500MG from Sigma Aldrich (St. Louis, MO).• Candida antarctica lipase B (CALB) Item # 62288-50MG-F from Sigma Aldrich (St. Louis, MO).

[0455] PCL and CALB enzymes were resuspended in ultra-pure water for stock concentrations of 81.4 U / ml and 3.95 U / ml respectively. Experimental groups consisted of formulation buffer without (-) PVP with (+) CALB, formulation buffer+PVP+CALB, formulation buffer-PVP+PCL, and formulation buffer+PVP+PCL.

[0456] For each experimental group, a master mix was prepared and 20 mL of the mix was aliquoted individually in five 20cc vials and immediately incubated at 5 °C until measured. Before measurements were taken, one vial was removed and placed on ice for each time point. For CALB and PCL, master mixes for both formulation buffers with and without PVP had the final CALB concentration of 0.01 U / ml and final PCL concentration of 12 U / ml with the remaining volume consisting of the respective buffer.

[0457] The enzyme-formulation buffer mixture samples were measured at 5 time points, 0 hrs, 1 hr, 3 hrs, 1 day, and 2 days after the master mixes were made. At each time point the samples were measured using Agilent Technologies 1260 Infinity HPLC equipped with an Agilent 1260 Infinity ELSD. Additionally, at each timepoint, samples were measured using the Liquid Particle Counter HIAC and inspected for visible particle formation.Results / Conclusion

[0458] Polysorbate 20 concentrations were measured at each time point by HPLC-ELSD and are shown in Table 14. The sample subvisible particle counts are shown in Table 15.91MF-363724269Attorney Docket No. 146392068840Table 14: Polysorbate 20 concentrations (mg / mL) measured by HPLC-ELSDTable 15: Subvisible particle counts > 2pm (particles per mL) measured by HIACSamples that exceeded the particle counting capabilities of the HIAC were noted as “Exceeded.” The samples taken on the final day for exceeded experimental groups were not taken and noted as not available (N / A).

[0459] FIG. 18 shows the polysorbate 20 concentrations normalized to their starting concentrations found in Table 14, subvisible particle counts in Table 15, and instances of observed visible particle formation. These data suggest that the presence of PVP does not accelerate FFA particle generation. The polysorbate 20 concentrations of FIG. 18 additionally show that the presence of PVP reduces the rate of polysorbate 20 degradation, most prevalently seen with PCL, an enzyme that primarily targets higher order esters. Despite there being no observed difference in onset of visible particle formation, a small difference in92MF-363724269Attorney Docket No. 146392068840 subvisible particle count was observed at the 1-day timepoint with CALB (boxed), an enzyme that primarily targets lower order esters. This lower subvisible particle count for the PVP case suggests that PVP has a protective effect against subvisible particle formation, only apparent with the enzyme with a slower polysorbate 20 degradation rate.

[0460] PVP has positive improvements for slowing PS20 degradation and subsequent subvisible particle formation.Example 10: Povidone molecular weight range 2000-11000 (polydispersity) is effective at solubilizing ocrelizumab SC under simulated physiological conditions (PBS at pH 7.2)Summary

[0461] The solubility of ocrelizumab subcutaneous (SC) high concentration (HC) formulated with different combination of Povidone K12 and Povidone K17 in 10 mM phosphate buffer saline (PBS) at pH 7.2 was assessed to study the effect of povidone molecular weight distribution (e.g., polydispersity) on the solubility of ocrelizumab SC HC. The results demonstrated no precipitation or insolubility of ocrelizumab SC HC when formulated with combinations of Povidone KI 2 and Povidone K17 after buffer exchanged into PBS at room temperature for 72 hours.

[0462] The ocrelizumab SC HC formulation composed of 110 mg / mL ocrelizumab formulated with 20 mM sodium acetate (As discussed above, due to the Gibbs-Donnan effect and volume excursion effects, the final sodium concentration is about 5 mM (in the range of 2 mM to 8 mM), and the final acetate concentration is about 43 mM (in the range of 37 mM to 47 mM) in the formulation), 180 mM sucrose, 30 mM methionine, 0.6 mg / mL polysorbate 20 at pH 5.3 with either povidone K12 or povidone K17 or mixtures of both at a final concentration of 75 mg / mL, were used in these experiments.Introduction

[0463] Povidone KI 2 (also known as polyvinylpyrrolidone or PvP KI 2) is added to ocrelizumab HC formulation to increase the ocrelizumab solubility at high concentrations when injected under the skin. Povidone K12 is polydisperse and has a weight average molecular weight of 2000 - 7000 Daltons. To study the effect of the polydispersity of povidone on the solubility of ocrelizumab SC HC in PBS (to mimic physiological environment), different mixtures of K12 and K17 were added in the formulation to achieve93MF-363724269Attorney Docket No. 146392068840 the final povidone concentration (75 mg / mL) in the formulation. Povidone K17 (also known as PvP K17) has a molecular weight range of 7,000 - 11,000 Daltons.Materials and Methods

[0464] In this investigative study, the ocrelizumab HC formulations with either PVP K12, PVP K17, or different mixtures of PVP K12 and PVP K17 in the formulation (Table 16) was buffer exchanged into PBS to assess the solubility.Table 16. Overview of Ocrelizumab High Concentration Formulations with different PVP K12, PVP K17, and Mixtures of BothResults

[0465] The appearance of the ocrelizumab HC solution formulated with different mixtures of PVP KI 2 and PVP K17 after dialysis into PBS using the 2 kD MWCO cassettes was visually assessed for any precipitation occurring at TO, 2 hours, 4 hours, 24 hours, 48 hours and 72 hours at room temperature during the buffer exchange. Pictures of the dialysis cassettes containing the buffer exchanged samples were taken and shown in FIG. 19A, FIG. 19B, FIG. 19C. The following was observed:1. At TO, after 2 or 4 hours of buffer exchange, no precipitation was observed in all formulations.2. After 24h, 48h, and 72h of buffer exchange, precipitation was observed in the positive control sample formulated without PVP (sample #1) only. Note that in sample #1 (positive control), precipitation was first observed after 24 hours of buffer exchange and confirmed after 48 hours of buffer exchange. The buffer exchange of sample #1 was terminated after 48 hours. No precipitation was observed in samples formulated with PVP12, PVP17, or the mixture over 72h.94MF-363724269Attorney Docket No. 146392068840Conclusion

[0466] These experiments confirm that PVP is able to solubilize ocrelizumab HC formulation in PBS. No precipitation was observed in all samples formulated with PVP(s) (either PVP K12 only, PVP K17 only, or a mixture of PVP K12 and PVP K17) and buffer exchanged into PBS at room temperature for up to 72 hours. The results demonstrated that the solubility of the ocrelizumab HC formulation is not affected by the molecular weight distribution (e.g., polydispersity) of povidone.Example 11: Ocrelizumab HC formulation

[0467] It is expected that in real- world manufacturing the actual concentration of ingredients in a formulation will differ slightly from the target concentration because of e.g., manufacturing variability. Typically observed concentration variation is within about 10% of the target. A long-term (at least 24-month) stability study is being conducted using the HC formulations that have various ingredients at concentrations that is at the upper or lower limits (see Table below).

[0468] As discussed above, due to the Gibbs-Donnan effect, because of the high concentration of the antibody and volume excursion effects during ultrafiltration and diafiltration (UF / DF) process, the final acetate concentration is about 43 mM (in the range of 37 mM to 47 mM) and the final sodium concentration is about 5 mM (in the range of 2 mM to 8 mM) in the formulation. The final sodium concentrations was measured using an ion- selective electrode method, while acetate concentration was determined using the Cedex Bio HT with acetate analysis.95MF-363724269Attorney Docket No. 14639206884096MF-363724269

Claims

Attorney Docket No. 146392068840CLAIMS1. An aqueous pharmaceutical formulation, the formulation comprising an anti- CD20 antibody in a concentration of about 99 mg / ml to about 121 mg / ml, hyaluronidase in a concentration of at least about 1000 U / ml, polyvinylpyrrolidone (PVP) KI 2 in a concentration of about 5% (w / v) to about 10% (w / v), methionine in a concentration of about 10 mM to about 50 mM, sucrose or trehalose, a surfactant, and a buffer, wherein the formulation has a pH of about 5.0 to about 5.6, wherein the anti-CD20 antibody comprises a VH domain comprising the amino acid sequence set forth in SEQ ID NO: 8, a VL domain comprising the amino acid sequence set forth in SEQ ID NO: 7, and a human IgGl constant region, wherein the formulation is suitable for subcutaneous administration.

2. The formulation of claim 1, wherein the anti-CD20 antibody is in a concentration of about 110 mg / ml.

3. The formulation of claim 1 or claim 2, wherein the PVP K12 is in a concentration of about 5% (w / v) to about 7.5% (w / v), optionally wherein the PVP K12 is in a concentration of about 7.5% (w / v).

4. The formulation of any one of claims 1-3, wherein the hyaluronidase is in a concentration of about 1000 U / ml to about 3000 U / ml, optionally 1500 U / ml to about 2500 U / ml, further optionally about 1800 U / ml to about 2200 U / ml, further optionally about 2000 U / ml.

5. The formulation of any one of claims 1-4, wherein methionine is in a concentration of about 10 mM to about 40 mM, optionally wherein methionine is in a concentration of 25 mM to about 35 mM, further optionally wherein methionine is in a concentration of about 30 mM.

6. The formulation of any one of claims 1-5, wherein the formulation comprises sucrose, optionally wherein the sucrose is in a concentration of about 150 mM to about 210 mM.97MF-363724269Attorney Docket No. 1463920688407. The formulation of claim 6, wherein the sucrose is in a concentration of about 180 mM.

8. The formulation of any one of claims 1-7, wherein the surfactant is polysorbate 20.

9. The formulation of claim 8, wherein the polysorbate is in a concentration of at least 0.03% (w / v), optionally wherein the polysorbate is in a concentration of about 0.03% (w / v) to about 0.08% (w / v) or about 0.04% (w / v) to about 0.08% (w / v).

10. The formulation of claim 9, wherein the polysorbate is in a concentration of about 0.06% (w / v).

11. The formulation of any one of claims 1-10, wherein the pH is about 5.3.

12. The formulation of any one of claims 1-11, wherein the buffer in the formulation is an acetate buffer.

13. The formulation of claim 12, wherein the acetate buffer has an acetate concentration of about 20 mM to about 50 mM.

14. The formulation of claim 13, wherein the acetate buffer has an acetate concentration of about 37 mM to about 47 mM, optionally wherein the acetate buffer has an acetate concentration of about 43 mM.

15. The formulation of any one of claims 1-14, wherein the buffer is a sodium acetate buffer.

16. The formulation of claim 15, wherein the sodium acetate buffer has a sodium concentration of about 2 mM to about 8 mM and an acetate concentration of about 37 mM to about 47 mM.

17. The formulation of claim 16, wherein the sodium acetate buffer has a sodium concentration of about 5 mM and an acetate concentration of about 43 mM.98MF-363724269Attorney Docket No. 14639206884018. An aqueous pharmaceutical formulation, the formulation comprising an anti- CD20 antibody in a concentration of 110 mg / ml, sucrose in a concentration of 180 mM, methionine in a concentration of 30 mM, polysorbate 20 in a concentration of 0.06% (w / v), hyaluronidase in a concentration of 2000 U / ml, polyvinylpyrrolidone (PVP) KI 2 in a concentration of 7.5% (w / v), and an acetate buffer, and wherein the formulation has a pH of 5.0 to about 5.6, wherein the anti-CD20 antibody comprises a VH domain comprising the amino acid sequence set forth in SEQ ID NO: 8, a VL domain comprising the amino acid sequence set forth in SEQ ID NO: 7, and a human IgGl constant region, wherein the formulation is suitable for subcutaneous administration.

19. The formulation of claim 18, wherein the formulation has a sodium acetate buffer, and wherein the sodium acetate buffer has a sodium concentration of 5 mM and an acetate concentration of 43 mM.

20. The formulation of any one of claims 1-19, wherein the weight average molecular weight of PVP KI 2 is 2000 Dalton to 7000 Dalton, optionally wherein the molecular weight is assessed by size exclusion chromatography (SEC) using light scattering detection, optionally wherein the weight average molecular weight of PVP K12 is 2000 Dalton to 3000 Dalton.

21. The formulation of any one of claims 1-20, wherein the anti-CD20 antibody comprises a light chain comprising the amino acid sequence of SEQ ID NO: 9 and a heavy chain comprising the amino acid sequence of SEQ ID NO: 10 or 11.

22. The formulation of any one of claims 1-21, wherein the anti-CD20 antibody is ocrelizumab.

23. The formulation of any one of claims 1-22, wherein the hyaluronidase is recombinant human hyaluronidase.

24. The formulation of any one of claims 1-23, wherein:99MF-363724269Attorney Docket No. 146392068840 a) the antibody in the formulation is stable at 2-8°C for at least 6 months, at least 12 months, at least 18 months or at least 24 months, b) the antibody and hyaluronidase in the formulation are stable after continuous agitation for 24 hours at room temperature, and / or c) there is i) less than 1% of aggregation of the antibody in the formulation as assessed by sum of HMW forms by SE-UHPLC after 25 days at 40°C and / or ii) less than 6% of degradation of the antibody in the formulation as measured by sum of pre-peaks by CE- SDS after 25 days at 40°C.

25. The formulation of claim 24, wherein the hyaluronidase in the formulation is stable when it retains at least about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of its biological activity.

26. The formulation of claim 24 or claim 25, wherein the antibody in the formulation is stable when it retains at least about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of its biological activity.

27. The formulation of claim 26, wherein the biological activity is measured by antibody binding to CD20.

28. The formulation of any one of claims 1-27, which is sterile.

29. A method of promoting solubility and bioavailability of an anti-CD20 antibody in an aqueous formulation, comprising adding a buffer comprising polyvinylpyrrolidone (PVP) K12 into a solution comprising the anti-CD20 antibody for preparation of the formulation, wherein the formulation comprises the antibody in a concentration of about 100 mg / ml to about 130 mg / ml and the PVP K12 in a concentration of about 7.5% (w / v).

30. The method of claim 29, wherein the formulation further comprises hyaluronidase.

31. A method of protecting an aqueous pharmaceutical formulation comprising an anti-CD20 antibody and hyaluronidase from agitation stress, comprising adding a buffer comprising polyvinylpyrrolidone (PVP) KI 2 into a solution comprising the anti-CD20100MF-363724269Attorney Docket No. 146392068840 antibody for preparation of the formulation, wherein the formulation comprises the PVP K12 in a concentration of at least about 5% (w / v).

32. The method of claim 31, wherein the PVP K12 has a concentration of between about 5% (w / v) and about 10% (w / v), optionally wherein the PVP K12 has a concentration of about 7.5% (w / v).

33. The method of any one of claims 30-32, wherein the hyaluronidase is in a concentration of at least about 1000 U / ml, optionally about 1000 U / ml to about 3000 U / ml, optionally about 1800 U / ml to about 2200 U / ml, further optionally wherein the hyaluronidase is in a concentration of about 2000 U / ml, further optionally wherein the hyaluronidase is a recombinant human hyaluronidase.

34. The method of any one of claims 29-33, wherein the anti-CD20 antibody is in a concentration of about 110 mg / ml.

35. The method of any one of claims 29-34, wherein the formulation further comprises methionine, optionally wherein methionine is in a concentration of about 10 mM to 50 mM, or about 10 mM to about 40 mM, or about 25 mM to about 35 mM, further optionally wherein methionine is in a concentration of about 30 mM.

36. The method of any one of claims 29-35, wherein the formulation comprises sucrose or trehalose.

37. The method of claim 36, wherein the formulation comprises sucrose, optionally wherein the formulation comprises sucrose in a concentration of about 150 mM to about 210 mM, further optionally wherein the sucrose is in a concentration of about 180 mM.

38. The method of any one of claims 29-37, wherein the surfactant is polysorbate, optionally polysorbate 20.

39. The method of claim 38, wherein the polysorbate is in a concentration of about 0.04% (w / v) to about 0.08% (w / v), optionally wherein the polysorbate is in a concentration of about 0.06% (w / v).101MF-363724269Attorney Docket No. 14639206884040. The method of any one of claims 29-39, wherein the pH is about 4.8 to about 5.8, optionally about 5.0 to about 5.6, further optionally 5.3.

41. The method of any one of claims 29-40, wherein the formulation comprises a sodium acetate buffer.

42. The method of claim 41, wherein the sodium acetate buffer has an acetate concentration of about 20 mM to about 50 mM.

43. The method of claim 41, wherein the acetate concentration is about 37 mM to about 47 mM, further optionally wherein the acetate concentration is 43 mM.

44. The method of claim 43, wherein the sodium acetate buffer has a sodium concentration of about 2 mM to about 8 mM, optionally wherein the sodium concentration is 5 mM.

45. The method of any one of claims 29-44, wherein the formulation comprises an anti-CD20 antibody in a concentration of about 110 mg / ml, sucrose in a concentration of about 180 mM, methionine in a concentration of about 30 mM, polysorbate 20 in a concentration of about 0.06% (w / v), hyaluronidase in a concentration of about 2000 U / ml, and PVP K12 in a concentration of about 7.5% (w / v), a sodium acetate buffer having a sodium concentration of about 5 mM and an acetate concentration of about 43 mM, and wherein the formulation has a pH of about 5.3.

46. The method of any one of claims 29-45, wherein the weight average molecular weight of PVP KI 2 is 2000 Dalton to 7000 Dalton, optionally wherein the molecular weight is assessed by size exclusion chromatography (SEC) using light scattering detection, optionally wherein the weight average molecular weight of PVP K12 is 2000 Dalton to 3000 Dalton.

47. The method of any one of claims 29-46, wherein the anti-CD20 antibody comprises a VH domain comprising the amino acid sequence set forth in SEQ ID NO: 8, a VL domain comprising the amino acid sequence set forth in SEQ ID NO: 7, and a human102MF-363724269Attorney Docket No. 146392068840IgGl constant region, and wherein the formulation is suitable for subcutaneous administration.

48. The method of claim 47, wherein the anti-CD20 antibody comprises a light chain comprising the amino acid sequence of SEQ ID NO: 9 and a heavy chain comprising the amino acid sequence of SEQ ID NO: 10 or 11.

49. A method of treating multiple sclerosis in a patient comprising subcutaneously administering the formulation of any one of claims 1-28.

50. The method of claim 49, wherein the patient has not been subjected to a prior intravenous administration of the anti-CD20 antibody at a dose of about 600 mg.

51. The method of claim 49 or claim 50, wherein the patient has not been subjected to a prior intravenous administration of the anti-CD20 antibody.

52. The method of any one of claims 49-51, wherein the patient has not been subjected to a prior subcutaneous administration of the anti-CD20 antibody at a dose lower than about 900 mg.

53. The method of claim 49, wherein the patient has received a prior treatment with an anti-CD20 antibody, optionally wherein the prior treatment comprises an intravenous administration of the anti-CD20 antibody, optionally wherein the prior treatment comprises the intravenous administration of the anti-CD20 antibody at a dose of about 300 mg or about 600 mg.

54. The method of any one of claims 49-53, wherein the method comprises subcutaneously administering more than one dose of the anti-CD20 antibody.

55. The method of claim 54, wherein the method comprises administering at least two, three or four doses of the anti-CD20 antibody.103MF-363724269Attorney Docket No. 14639206884056. The method of any one of claims 49-55, wherein the anti-CD20 antibody is administered at a frequency of no more than once every 24 weeks or about once every 24 weeks.

57. The method of any one of claims 49-56, wherein the anti-CD20 antibody is administered at a frequency of no more than once every 6 months or about once every 6 months.

58. The method of any one of claims 49-57, wherein anti-CD20 antibody is the only medicament administered to the patient to treat multiple sclerosis.

59. The method of any one of claims 49-58, wherein the patient is subject to a) an oral dexamethasone or an equivalent corticosteroid and b) an oral antihistamine administration (e.g., desloratadine) shortly prior to the subcutaneous administration of the anti-CD20 antibody, optionally wherein the oral dexamethasone or the equivalent corticosteroid and antihistamine are administered at least or within 30 minutes, 20 minutes, or 15 minutes prior to the subcutaneous administration, further optionally wherein the patient is subject to 20 mg dexamethasone and 5 mg desloratadine.

60. The method of any one of claims 49-59, wherein the multiple sclerosis is relapsing multiple sclerosis (RMS).

61. The method of any one of claims 49-60, wherein the patient has a clinical isolated syndrome (CIS), relapsing-remitting multiple sclerosis (RRMS) or active secondary progressive multiple sclerosis (SPMS).

62. The method of any one of claims 49-61, wherein the multiple sclerosis is a primary progressive multiple sclerosis (PPMS).

63. The method of any one of claims 49-62, wherein the patient has an Expanded Disability Status Scale (EDSS) score of 0-6.5.

64. The method of any one of claims 49-63, wherein the subcutaneous administration takes no more than about 10 minutes.104MF-363724269Attorney Docket No. 14639206884065. The method of any one of claims 49-64, wherein said anti-CD20 antibody in the formulation is about 110 mg / ml.

66. The method of any one of claims 49-65, wherein the anti-CD20 antibody comprises a light chain comprising the amino acid sequence of SEQ ID NO: 9 and a heavy chain comprising the amino acid sequence of SEQ ID NO: 10 or 11.

67. The method of any one of claims 49-66, wherein the anti-CD20 antibody is ocrelizumab.105MF-363724269

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