Compositions and methods for treating cancer using subcutaneous administration of anti-PD1 antibodies

Abstract

The present invention relates to compositions and methods for treating cancer in a patient comprising administering to the patient subcutaneously every six weeks a specific amount of a PD-1 antagonist, e.g., an anti-PD-1 antibody (e.g., pemmumab) or an antigen-binding fragment thereof, with or without a hyaluronidase. In particular embodiments, the amount of the anti-PD-1 antibody or antigen-binding fragment thereof is from about 600 mg to about 1000 mg. In particular embodiments, the administration is performed once every about three weeks, and the amount of the anti-PD-1 antibody or antigen-binding fragment thereof is from about 300 mg to about 500 mg. In certain embodiments, the PD-1 antagonist is pembromab or an antigen binding fragment thereof. Also provided are compositions and kits formulated for subcutaneous administration comprising specific doses of an anti-PD-1 antibody or antigen-binding fragment thereof, and uses thereof for the treatment of cancer.

Description

[0001] Cross - Reference to Related Applications

[0002] This application is an international patent application that claims priority and the benefit of U.S. Provisional Application No. 63 / 449,478, filed on March 2, 2023, U.S. Provisional Application No. 63 / 415,928, filed on October 13, 2022, and U.S. Provisional Application No. 63 / 415,526, filed on October 12, 2022, the entire contents of each of which are incorporated herein by reference. Technical Field

[0003] The present invention relates to compositions and therapies that can be used to treat cancer. In particular, the present invention relates to a method of treating cancer that includes administering an anti-PD-1 antibody or an antigen-binding fragment thereof to a patient in need thereof using the dosing regimens specified herein. Compositions and kits formulated for subcutaneous administration are also provided, which contain a specific dose of an anti-PD-1 antibody or an antigen-binding fragment thereof. Background Art

[0004] PD-1 is considered an important player in immune regulation and maintenance of peripheral tolerance. PD-1 is moderately expressed on naive T, B, and natural killer T (NKT) cells and is upregulated by T / B cell receptor signaling on lymphocytes, monocytes, and myeloid cells (Sharpe et al., The function of programmed cell death 1 and its ligands in regulating autoimmunity and infection. Nature Immunology (2007); 8:239-245).

[0005] The two known ligands of PD-1, PD-L1 (B7-H1) and PD-L2 (B7-DC), are expressed in human cancers that occur in a variety of tissues. In large cohorts of cancers (e.g., ovarian cancer, renal cancer, colorectal cancer, pancreatic cancer, and liver cancer and melanoma), it has been shown that PD-L1 expression is associated with poor prognosis and shortened overall survival regardless of subsequent treatment (Dong et al., Nat Med. 8(8):793-800 (2002); Yang et al., Invest Ophthalmol Vis Sci. 49:2518-2525 (2008); Ghebeh et al., Neoplasia 8:190-198 (2006); Hamanishi et al., Proc. Natl. Acad. Sci. USA 104:3360-3365 (2007); Thompson et al., Cancer 5:206-211 (2006); Nomi et al., Clin. Cancer Research 13:2151-2157 (2007); Ohigashi et al., Clin. Cancer Research 11:2947-2953 (2005); Inman et al., Cancer 109:1499-1505 (2007); Shimauchi et al., Int. J. Cancer 121:2585-2590 (2007); Gao et al., Clin. Cancer Research 15:971-979 (2009); Nakanishi J. Cancer Immunol Immunother. 56:1173-1182 (2007); and Hino et al., Cancer 00:1-9 (2010)).

[0006] Similarly, the expression marker of PD-1 on tumor-infiltrating lymphocytes has been found to be dysfunctional T cells in breast cancer and melanoma (Ghebeh et al., BMC Cancer. 20088:5714-15 (2008); and Ahmadzadeh et al., Blood 114:1537-1544 (2009)) and is associated with a poor prognosis in renal cancer (Thompson et al., Clinical Cancer Research 15:1757-1761 (2007)). Thus, it has been proposed that tumor cells expressing PD-L1 interact with T cells expressing PD-1 to attenuate T cell activation and evade immune surveillance, resulting in impaired immune responses to tumors.

[0007] Immune checkpoint therapies targeting the PD-1 axis have made breakthrough progress in the clinical responses of many human cancers (Brahmer et al., N Engl J Med 2012, 366:2455-65; Garon et al., N Engl J Med 2015, 372:2018-28; Hamid et al., N Engl J Med 2013, 369:134-44; Robert et al., Lancet 2014, 384:1109-17; Robert et al., N Engl J Med 2015, 372:2521-32; Robert et al., N Engl J Med 2015, 372:320-30; Topalian et al., N Engl J Med 2012, 366:2443-54; Topalian et al., J Clin Oncol 2014, 32:1020-30; and Wolchok et al., N Engl J Med 2013, 369:122-33). Immune therapies targeting the PD-1 axis include monoclonal antibodies against the PD-1 receptor (KEYTRUDA TM (pembrolizumab), Merck and Co., Inc., Kenilworth, NJ, USA and OPDIVO TM (nivolumab), Bristol-Myers Squibb Company, Princeton, NJ, USA), and those that bind to the PD-L1 ligand (MPDL3280A; TECENTRIQ TM (atezolizumab), Genentech, San Francisco, CA, USA; IMFINZI TM (durvalumab), AstraZeneca Pharmaceuticals LP, Wilmington, DE; and BAVENCIO TM (avelumab), Merck KGaA, Darmstadt, Germany). Both of these treatment methods have proven to have anti-tumor effects in many cancer types.

[0008] Hyaluronidase is an enzyme that degrades hyaluronic acid present in the extracellular matrix. There are six types of hyaluronidase known in humans: Hyal1, Hyal2, Hyal3, Hyal4, HyalPS1, and PH20 / SPAM1. PH20 / SPAM1 (hereinafter referred to as PH20) is expressed in the sperm plasma membrane and acrosomal membrane.

[0009] Hyaluronidase hydrolyzes hyaluronic acid, thereby reducing the viscosity of hyaluronic acid in the extracellular matrix and increasing its permeability into tissues (skin). The subcutaneous region of the skin has a neutral pH of approximately 7.0 to 7.5. Among various types of hyaluronidases, PH20 is widely used (Bookbinder et al., 2006). In instances where PH20 is used, PH20 is typically co-administered with an antibody therapeutic agent by subcutaneous injection (Bookbinder et al., 2006). rHuPH20 (also known as ), approved by the FDA, is designated as an adjuvant to enhance the dispersion and absorption of other injectable drugs.

[0010] Currently approved anti-PD-1 antibody therapies for multiple cancer indications are administered by IV infusion at a dose of (i) 200 mg or 2 mg / kg Q3W or (ii) 400 mg Q6W. Developing a dosing regimen that allows for the safe and effective subcutaneous dosing of anti-PD-1 antibodies with hyaluronidase, which provides exposure comparable to the approved IV infusion dose, would be beneficial. An alternative to IV infusion, such as subcutaneous administration, would provide convenience and flexibility to patients, reduce the time patients spend in the treatment room, and shorten the time required for the provider to administer the treatment. SUMMARY OF THE INVENTION

[0011] The present invention provides an alternative, convenient, cost-effective subcutaneous dosing regimen for treating cancer patients with an anti-PD-1 antibody or an antigen-binding fragment thereof, wherein the dosing regimen is expected to provide a safe and effective dose of the anti-PD-1 antibody or an antigen-binding fragment thereof. Specifically, the present invention provides a method for treating cancer in a human patient in need thereof, comprising subcutaneously administering to the patient a dose of from about 600 mg to about 1000 mg of an anti-PD-1 antibody or an antigen-binding fragment thereof every six weeks; wherein the anti-PD-1 antibody or an antigen-binding fragment thereof comprises (a) light chain (LC) complementarity-determining regions (CDRs) LC-CDR1, LC-CDR2, and LC-CDR3, which comprise the amino acid sequences shown in SEQ ID NOs: 1, 2, and 3, respectively, and heavy chain (HC) CDRs HC-CDR1, HC-CDR2, and HC-CDR3, which comprise the amino acid sequences shown in SEQ ID NOs: 6, 7, and 8, respectively. In certain embodiments, the antibody or an antigen-binding fragment thereof is co-administered with hyaluronidase, and in certain embodiments, the hyaluronidase comprises human hyaluronidase. In an embodiment of the present invention, the antibody or an antigen-binding fragment thereof is pembrolizumab or an antigen-binding fragment thereof. In a further embodiment, the anti-PD-1 antibody is pembrolizumab.

[0012] The present invention also provides a method for treating cancer in a human patient in need thereof, which comprises subcutaneously administering to the patient a dose of about 300 mg to about 500 mg of an anti-PD-1 antibody or an antigen-binding fragment thereof and hyaluronidase about every three weeks; wherein the anti-PD-1 antibody or an antigen-binding fragment thereof comprises (a) light chain (LC) complementarity determining regions (CDRs) LC-CDR1, LC-CDR2 and LC-CDR3, which comprise the amino acid sequences shown in SEQ ID NOs: 1, 2 and 3, respectively, and heavy chain (HC) CDRs HC-CDR1, HC-CDR2 and HC-CDR3, which comprise the amino acid sequences shown in SEQ ID NOs: 6, 7 and 8, respectively. In an embodiment of the present invention, the hyaluronidase is human hyaluronidase. In an embodiment of the present invention, the antibody or antigen-binding fragment is pembrolizumab or an antigen-binding fragment thereof. In a further embodiment, the anti-PD-1 antibody is pembrolizumab.

[0013] The present invention also relates to a pharmaceutical composition for subcutaneous injection, which comprises a dose of about 600 mg to about 1000 mg of an anti-PD-1 antibody or an antigen-binding fragment thereof, and about 700 units to about 50000 units of human hyaluronidase. The present invention also relates to a pharmaceutical composition for subcutaneous injection, which comprises a dose of about 300 mg to about 500 mg of an anti-PD-1 antibody or an antigen-binding fragment thereof, and about 700 units to about 50000 units of human hyaluronidase.

[0014] In all the treatment methods, compositions and uses disclosed herein, the anti-PD-1 antibody or antigen-binding fragment inhibits the binding of PD-L1 to PD-1, and preferably also inhibits the binding of PD-L2 to PD-1. In a specific embodiment of the treatment methods, compositions and uses of the present invention, the anti-PD-1 antibody or antigen-binding fragment is a monoclonal antibody that specifically binds to PD-1 and blocks the binding of PD-L1 to PD-1. In a specific embodiment, the anti-PD-1 antibody comprises a heavy chain and a light chain, wherein the light chain and heavy chain comprise the amino acid sequences of SEQ ID NO: 5 and SEQ ID NO: 10 or 11, respectively.

[0015] In a specific embodiment of the disclosed treatment methods, compositions and uses, the cancer expresses one or both of PD-L1 and PD-L2. In a specific embodiment, PD-L1 expression is present or elevated in the cancer. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 Shows C infused IV at 200 mg Q3W in KEYNOTE-252 troughFlat exposure-response comparison of tumor size change. C from KEYNOTE-555 400 mg Q6W IV infusion trough Falling into the C of establishing a flat exposure-response trough within this range.

[0017] Figure 2A -F. Comparison of observed pharmacokinetic data from KEYNOTE-555 with model-predicted pharmacokinetic properties for pembrolizumab 400 mg Q6W (A: Cycle 1, B: Steady State); Model-predicted pharmacokinetic properties for pembrolizumab 200 mg Q3W and 2 mg / kg Q3W (C) trough Comparison with 400 mg Q6W (C: initial treatment for six weeks, D: steady state); and model predictions for pembrolizumab 200 mg Q3W, 2 mg / kg Q3W, and 10 mg / kg Q2W C trough Comparison with 400 mg Q6W (E: initial treatment for six weeks, F: steady state).

[0018] Figure 3A Simulations based on a pharmacokinetic ("PK") model at cycle 1 for pembrolizumab 790 mg Q6W SC and 400 mg Q6W IV doses are shown. trough Distribution (5th, 25th, 50th, 75th, and 95th percentiles; solid lines from bottom to top). Minimum IV refers to the 5th percentile value of the exposure index distribution from the 400 mg Q6W IV dose; Median IV refers to the 50th percentile value of the exposure index distribution from the 400 mg Q6W IV dose; Maximum IV refers to the 95th percentile value of the exposure index distribution from the 400 mg Q6W IV dose; Minimum IV, Median IV, and Maximum IV are dashed lines from bottom to top.

[0019] Figure 3B The C values ​​of 790 mg Q6W SC and 400 mg Q6W IV doses of pembrolizumab at steady state (cycle 3) using PK model-based simulations are shown. trough Distribution (5th, 25th, 50th, 75th, and 95th percentiles; solid lines from bottom to top). Minimum IV refers to the 5th percentile value of the exposure index distribution from the 400 mg Q6W IV dose; Median IV refers to the 50th percentile value of the exposure index distribution from the 400 mg Q6W IV dose; Maximum IV refers to the 95th percentile value of the exposure index distribution from the 400 mg Q6W IV dose; Minimum IV, Median IV, and Maximum IV are dashed lines from bottom to top.

[0020] Figure 4AShows the AUC at Cycle 1 using PK model-based simulations for pembrolizumab 790 mg Q6W SC and 400 mg Q6W IV doses 0-6周 Distribution (5th, 25th, 50th, 75th, and 95th percentiles; solid lines from bottom to top). Min IV refers to the 5th percentile value of the exposure metric distribution from the 400 mg Q6W IV dose; Median IV refers to the 50th percentile value of the exposure metric distribution from the 400 mg Q6W IV dose; Max IV refers to the 95th percentile value of the exposure metric distribution from the 400 mg Q6W IV dose; Min IV, Median IV, and Max IV are dashed lines from bottom to top.

[0021] Figure 4B Shows the AUC at steady state (Cycle 3) using PK model-based simulations for pembrolizumab 790 mg Q6W SC and 400 mg Q6W IV doses 0-6周 Distribution (5th, 25th, 50th, 75th, and 95th percentiles; solid lines from bottom to top). Min IV refers to the 5th percentile value of the exposure metric distribution from the 400 mg Q6W IV dose; Median IV refers to the 50th percentile value of the exposure metric distribution from the 400 mg Q6W IV dose; Max IV refers to the 95th percentile value of the exposure metric distribution from the 400 mg Q6W IV dose; Min IV, Median IV, and Max IV are dashed lines from bottom to top.

[0022] Figure 5 Shows the C at steady state (Cycle 3) using PK model-based simulations for pembrolizumab 790 mg Q6W SC and 400 mg Q6W IV doses max Distribution (5th, 25th, 50th, 75th, and 95th percentiles; solid lines from bottom to top). Min IV refers to the 5th percentile value of the exposure metric distribution from the 400 mg Q6W IV dose; Median IV refers to the 50th percentile value of the exposure metric distribution from the 400 mg Q6W IV dose; Max IV refers to the 95th percentile value of the exposure metric distribution from the 400 mg Q6W IV dose; Min IV, Median IV, and Max IV are dashed lines from bottom to top.

[0023] Figure 6A Shows the C at Cycle 1 using PK model-based simulations for pembrolizumab-HLN at 395 mg Q3W (left panel is C after 3 weeks trough ; right panel is C after 6 weeks trough ) and 790 mg Q6W SC doses troughDistribution (5th, 25th, 50th, 75th, and 95th percentiles; solid lines from bottom to top). Min Ref refers to the 5th percentile value of the exposure metric distribution from the 790 mg Q6W SC dose; Median Ref refers to the 50th percentile value of the exposure metric distribution from the 790 mg Q6W SC dose; Max Ref refers to the 95th percentile value of the exposure metric distribution from the 790 mg Q6W SC dose; Min Ref, Median Ref, and Max Ref are dashed lines from bottom to top.

[0024] Figure 6B Shows C at steady state using PK model-based simulations at doses of 395 mg Q3W and 790 mg Q6W SC of pembrolizumab-HLN trough Distribution (5th, 25th, 50th, 75th, and 95th percentiles; solid lines from bottom to top). Min Ref refers to the 5th percentile value of the exposure metric distribution from the 790 mg Q6W SC dose; Median Ref refers to the 50th percentile value of the exposure metric distribution from the 790 mg Q6W SC dose; Max Ref refers to the 95th percentile value of the exposure metric distribution from the 790 mg Q6W SC dose; Min Ref, Median Ref, and Max Ref are dashed lines from bottom to top.

[0025] Figure 7A Shows AUC at Cycle 1 using PK model-based simulations at doses of 395 mg Q3W and 790 mg Q6W SC of pembrolizumab-HLN 0-6周 Distribution (5th, 25th, 50th, 75th, and 95th percentiles; solid lines from bottom to top). Min Ref refers to the 5th percentile value of the exposure metric distribution from the 790 mg Q6W SC dose; Median Ref refers to the 50th percentile value of the exposure metric distribution from the 790 mg Q6W SC dose; Max Ref refers to the 95th percentile value of the exposure metric distribution from the 790 mg Q6W SC dose; Min Ref, Median Ref, and Max Ref are dashed lines from bottom to top.

[0026] Figure 7B Shows AUC at steady state using PK model-based simulations at doses of 395 mg Q3W and 790 mg Q6W SC of pembrolizumab-HLN 0-6周Distribution (5th, 25th, 50th, 75th, and 95th percentiles; solid line from bottom to top). Min Ref refers to the 5th percentile value of the exposure metric distribution from the 790 mg Q6W SC dose; Median Ref refers to the 50th percentile value of the exposure metric distribution from the 790 mg Q6W SC dose; Max Ref refers to the 95th percentile value of the exposure metric distribution from the 790 mg Q6W SC dose; Min Ref, Median Ref, and Max Ref are dashed lines from bottom to top.

[0027] Figure 8 is the similarity of pembrolizumab clearance based on the PK model between different indications. The distribution percentiles of the post - hoc estimated individual baseline clearance rate values in subjects for each indication (number of subjects for each indication shown above) are represented by the line (50th percentile), box (25th - 75th percentile), and whiskers (5th - 95th percentile). The sample size (N) for each group is provided in each of the above box - whisker plots. NSCLC = non - small cell lung cancer; HN = head and neck squamous cell carcinoma; UC = urothelial carcinoma; MSIH = high microsatellite instability cancer; HCC = hepatocellular carcinoma; cHL = classical Hodgkin lymphoma; PMBCL = primary mediastinal B - cell lymphoma. Detailed Description

[0028] The present invention provides a method of treatment (e.g., a method of treating cancer) for a patient (e.g., a human patient), which comprises subcutaneously administering a specified dose of an anti - PD - 1 antibody (e.g., pembrolizumab) or an antigen - binding fragment thereof, and optionally hyaluronidase (and in certain embodiments, human hyaluronidase). Such administration is expected to provide a safe and effective dose of the anti - PD - 1 antibody or an antigen - binding fragment thereof. Also provided are compositions and kits formulated for subcutaneous administration, which comprise a dose of an anti - PD - 1 antibody or an antigen - binding fragment thereof, and optionally hyaluronidase (e.g., human hyaluronidase), and their use for treating cancer. In certain embodiments of the present invention, the anti - PD - 1 antibody is pembrolizumab or an antigen - binding fragment of pembrolizumab.

[0029] I. Abbreviations and Definitions

[0030] When used throughout the specification and the appended claims, the following abbreviations apply:

[0031] AUC Area Under the Concentration - Time Curve

[0032] AUCss Area Under the Concentration - Time Curve at Steady State

[0033] CDR Complementary Determining Region

[0034] Cl Confidence Interval

[0035] CL clearance rate

[0036] C max,ss Peak concentration at steady state

[0037] CPS combined positive score

[0038] CV coefficient of variation of the inter-subject distribution of the parameter;

[0039] ECOG Eastern Cooperative Oncology Group

[0040] eGFR: estimated glomerular filtration rate

[0041] E-R exposure (concentration)-response

[0042] F bioavailability

[0043] FFPE formalin-fixed paraffin-embedded

[0044] FR framework region

[0045] GM geometric mean

[0046] HCC hepatocellular carcinoma

[0047] HNSCC head and neck squamous cell carcinoma

[0048] HL Hodgkin lymphoma

[0049] IgG immunoglobulin G

[0050] IHC immunohistochemistry or immunohistochemical

[0051] IMAX: maximum effect of time on CL

[0052] IV intravenous

[0053] ka first-order absorption rate constant

[0054] LPS lymphoma proportion score

[0055] mAb monoclonal antibody

[0056] MCC Merkel cell carcinoma

[0057] MEL melanoma

[0058] MMR mismatch repair

[0059] MPS modified proportion score

[0060] MRI magnetic resonance imaging

[0061] MSI-H high microsatellite instability

[0062] National Cancer Institute (NCI)

[0063] Non-small cell lung cancer (NSCLC)

[0064] Overall survival (OS)

[0065] Programmed death 1 (also known as programmed cell death-1 and programmed death receptor 1) (PD-1)

[0066] Programmed death ligand 1 (PD-L1)

[0067] Programmed death ligand 2 (PD-L2)

[0068] Progression-free survival (PFS)

[0069] Pharmacokinetics (PK)

[0070] Clearance (CL)

[0071] Once every two weeks (Q2W)

[0072] Once every three weeks (Q3W)

[0073] Once every six weeks (Q6W)

[0074] Renal cell carcinoma (RCC)

[0075] Relative standard error (RSE)

[0076] Subcutaneous (SC)

[0077] TI 50 Time to achieve 50% of the maximum effect on clearance

[0078] t lag Absorption lag time

[0079] Tumor proportion score (TPS)

[0080] Central volume of distribution (Vc)

[0081] V H Variable region of immunoglobulin heavy chain

[0082] V L Variable region of immunoglobulin light chain

[0083] Peripheral volume of distribution (Vp)

[0084] The presented population parameter estimates exclude the covariate effects; thus, these estimates apply to a hypothetical typical patient with average characteristics.

[0085] To more readily understand the present invention, certain technical and scientific terms are specifically defined below. Unless otherwise specifically defined elsewhere in this document, all other technical and scientific terms used herein have the meaning commonly understood by one of ordinary skill in the art to which this invention pertains.

[0086] As used throughout the specification and the appended claims, unless the context clearly dictates otherwise, the singular forms "a," "an," and "the" include plural references. References to "or" mean one or both of the alternatives, unless the context clearly dictates otherwise. In some instances, the phrase "and / or" is used to emphasize one or both of the alternatives.

[0087] When modifying the amount of a substance or composition (e.g., mg) or the value of a parameter representing a method step, etc., the term "about" or "approximately" refers to the amount variations that can occur through typical measurement, handling, and sampling procedures involved in the preparation, characterization, and / or use of the substance or composition; through inadvertent errors in these procedures; through differences in the manufacture, source, or purity of the ingredients used to prepare or use the composition or perform the procedure; and so on. In certain embodiments, "about" or "approximately" refers to a variation of ±0.1%, ±0.5%, ±1%, ±2%, ±3%, ±4%, ±5%, ±6%, ±7%, ±8%, ±9%, ±10%, or ±11%.

[0088] When applied to animals, humans, experimental subjects, cells, tissues, organs, or biological fluids, "administering" and "treating" refer to the contact of an exogenous drug, therapeutic agent, diagnostic agent, or composition with an animal, human, subject, cell, tissue, organ, or biological fluid. As used herein, "treating" or "treatment" of cancer refers to administering an anti-PD-1 antibody or antigen-binding fragment to a subject having or diagnosed with cancer to achieve at least one positive therapeutic effect on cancer, such as, for example, reducing the number of cancer cells, shrinking the tumor size, decreasing the rate of cancer cell infiltration into peripheral organs, or decreasing the rate of tumor metastasis or tumor growth. "Treatment" can include one or more of the following: inducing / increasing an anti-tumor immune response, reducing the amount of one or more tumor markers, halting or delaying the growth of a tumor or blood cancer or the progression of a PD-1-related disease (a "PD-1-related disease") (such as cancer) associated with the binding of PD-1 to its ligands PD-L1 and / or PD-L2, stabilizing a PD-1-related disease, inhibiting the growth or survival of tumor cells, eliminating or shrinking the size of one or more cancer lesions or tumors, reducing the level of one or more tumor markers, improving or eliminating the clinical manifestations of a PD-1-related disease, decreasing the severity or duration of the clinical symptoms of a PD-1-related disease (such as cancer), prolonging the survival of a patient relative to the expected survival of a similar untreated patient, and inducing a complete or partial remission of cancer or other PD-1-related diseases.

[0089] The positive therapeutic effects of cancer can be measured in a variety of ways (see W.A. Weber, J. Nucl. Med. 50:1S-10S (2009)). For example, for tumor growth inhibition, according to NCI criteria, T / C ≤ 42% is the minimum level of anti-tumor activity. A T / C < 10% is considered a high level of anti-tumor activity, where T / C (%) = median tumor volume in treated / median tumor volume in control × 100. In certain embodiments, the treatment achieved by a therapeutically effective amount is any one of progression-free survival (PFS), disease-free survival (DFS), or overall survival (OS). PFS, also known as "time to tumor progression," refers to the length of time during and after treatment that cancer does not grow and includes the amount of time a patient experiences a complete or partial remission, as well as the amount of time a patient experiences disease stabilization. DFS refers to the length of time during and after treatment that a patient remains disease-free. OS refers to an extended life expectancy compared to an untreated individual or patient at the initial time. Although embodiments of the methods, compositions, and uses of the present invention may not effectively achieve a positive therapeutic effect in every patient, they should achieve a positive therapeutic effect in a statistically significant number of subjects, such as by any statistical test known in the art (such as Student's t-test, chi 2determined by tests, the Mann-Whitney U test, the Kruskal-Wallis test (H test), the Jonckheere-Terpstra test, and the Wilcoxon test).

[0090] "Antibody" refers to any form of antibody that exhibits the desired biological or binding activity. Thus, it is used in the broadest sense and specifically encompasses, but is not limited to, monoclonal antibodies (including full-length monoclonal antibodies), polyclonal antibodies, humanized, fully human antibodies, and chimeric antibodies. A "parent antibody" is an antibody obtained by exposing the immune system to an antigen prior to modifying the antibody for its intended use (e.g., humanizing the antibody for use as a human therapeutic).

[0091] Under normal circumstances, the basic antibody structural unit consists of a tetramer. Each tetramer contains two pairs of identical polypeptide chains, each pair having one "light" chain (about 25 kDa) and one "heavy" chain (about 50 - 70 kDa). The amino terminus of each chain contains a variable region of about 100 to 110 or more amino acids that is primarily responsible for antigen recognition. The carboxyl-terminal portion of the heavy chain can define a constant region that is primarily responsible for effector functions. Generally, human light chains are classified as κ and λ light chains. In addition, human heavy chains are typically divided into μ, δ, γ, α, or ε, and the isotypes of the antibody are defined as IgM, IgD, IgG, IgA, and IgE, respectively. In both the light and heavy chains, the variable and constant regions are joined by a "J" region of about 12 or more amino acids, and the heavy chain also contains a "D" region of about 10 or more amino acids. Generally, see, Fundamental Immunology Ch. 7 (Paul, W. ed., 2nd ed., Raven Press, N.Y. (1989)).

[0092] The variable regions of each light / heavy chain pair form the antibody binding site. Thus, under normal circumstances, an intact antibody has two binding sites. Except for bifunctional or bispecific antibodies, under normal circumstances, the two binding sites are identical.

[0093] Generally, both the variable domains of the heavy and light chains contain three hypervariable regions, also known as complementarity-determining regions (CDRs), which are located within relatively conserved framework regions (FRs). The CDRs are usually arranged by the framework regions such that they can bind to a specific epitope. Under normal circumstances, from the N-terminus to the C-terminus, both the light and heavy chain variable domains contain FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4. The assignment of amino acids to each domain is usually based on Sequences of Proteins of Immunological InterestDefinition by Kabat et al., National Institutes of Health, Bethesda, Md., 5th Edition, NIH Publication No. 91-3242 (1991), Kabat (1978) Adv. Prot. Chem. 32:1-75, Kabat, et al., (1977) J. Biol. Chem. 252:6609-6616, Chothia, et al., (1987) J Mol. Biol. 196:901-917 or Chothia et al., (1989) Nature 342:878-883.

[0094] Unless otherwise specified, "antibody fragment" or "antigen-binding fragment" refers to an antigen-binding fragment of an antibody, i.e., an antibody fragment that retains the ability to specifically bind to the antigen bound by the full-length antibody, e.g., a fragment that retains one or more CDR regions, e.g., three heavy-chain CDRs and three light-chain CDRs. Examples of antigen-binding fragments include, but are not limited to, Fab, Fab', F(ab')2, and Fv fragments.

[0095] As used in any of the therapeutic methods, compositions, and uses of the present invention, "anti-PD-1 antibody" includes a monoclonal antibody (mAb) or an antigen-binding fragment thereof that specifically binds to human PD-1. Alternative names or synonyms for PD-1 and its ligands include: PDCD1, PD1, CD279, and SLEB2 for PD-1; PDCD1L1, PDL1, B7H1, B7-4, CD274, and B7-H for PD-L1; and PDCD1L2, PDL2, B7-DC, Btdc, and CD273 for PD-L2. In any of the therapeutic methods, compositions, and uses of the present invention in which a human individual is being treated, the anti-PD-1 antibody or an antigen-binding fragment thereof is a PD-1 antagonist that blocks the binding of human PD-L1 to human PD-1 or blocks the binding of both human PD-L1 and PD-L2 to human PD-1. The amino acid sequence of human PD-1 can be found at NCBI locus number: NP_005009. The amino acid sequences of human PD-L1 and PD-L2 can be found at NCBI locus numbers: NP_054862 and NP_079515, respectively. The anti-PD-1 antibody can be a human antibody, a humanized antibody, or a chimeric antibody, and can comprise a human constant region. In a particular embodiment, the human constant region is selected from the following: IgG1, IgG2, IgG3, and IgG4 constant regions, and in a particular embodiment, the human constant region is an IgG1 or IgG4 constant region. In a particular embodiment, the antigen-binding fragment is selected from the following: Fab, Fab'-SH, F(ab')2, scFv, and Fv fragments.

[0096] "AUC" and "Cmax "is a pharmacokinetic parameter that reflects the systemic exposure to a drug (e.g., pembrolizumab) in the human body after its administration, and is generally considered a driver of drug efficacy. "AUC" represents the average exposure within the dosing interval. "C max " is the maximum or highest (peak) drug concentration observed shortly after its administration. In the specific case of pembrolizumab administered subcutaneously, the peak concentration occurs immediately after the end of the infusion. C is generally considered max a measure of the safety driver.

[0097] "Biotherapeutic agent" refers to a biomolecule that blocks ligand / receptor signal transduction in any biological pathway that supports tumor maintenance and / or growth or inhibits the anti-tumor immune response, such as an antibody or a fusion protein.

[0098] The term "buffer" encompasses those reagents that maintain the solution pH of the formulations of the present invention within an acceptable range, or for the lyophilized formulations of the present invention, provide an acceptable solution pH prior to lyophilization. The terms "lyophilize", "lyophilized" and "freeze-dry" refer to the process of first freezing the material to be dried and then removing ice or the frozen solvent by sublimation in a vacuum environment. Excipients can be added to the pre-lyophilized formulations to improve the stability of the lyophilized product during storage.

[0099] "C trough " is the trough concentration reached at the end of the dosing interval. SC:IV C trough Ratio is the C trough ratio (e.g., geometric mean ratio) reached using the SC dose relative to the IV dose at the end of the same dosing interval.

[0100] As used herein, "co-administer" refers to agents administered to a subject simultaneously or at about the same time. The agents may or may not be physically combined before administration. For example, an anti-PD-1 antibody and hyaluronidase may be in separate vials and, when in a liquid solution, can be mixed into the same injection device and administered to a patient simultaneously.

[0101] As used herein, "co-formulated" or "co-formulation" or "coformulation" or "co-formulated" refers to at least two different proteins or agents formulated together and stored in a single vial, container, device or vessel (e.g., an injection device) as a combination product rather than being formulated and stored separately and then mixed or administered separately before administration.

[0102] The terms "cancer", "cancerous", or "malignant" refer to or describe a physiological condition in a mammal that is typically characterized by uncontrolled cell growth. Examples of cancers include, but are not limited to, carcinomas, lymphomas, leukemias, blastomas, and sarcomas. More specific examples of such cancers include, but are not limited to, squamous cell carcinoma, myeloma, small cell lung cancer, non-small cell lung cancer, glioma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, acute myeloid leukemia (AML), multiple myeloma, gastrointestinal (GI) cancer, renal cancer, ovarian cancer, liver cancer, lymphoblastic leukemia, lymphocytic leukemia, colorectal cancer, endometrial cancer, kidney cancer, prostate cancer, thyroid cancer, melanoma, chondrosarcoma, neuroblastoma, pancreatic cancer, glioblastoma multiforme, cervical cancer, brain cancer, gastric cancer, bladder cancer, liver cancer, breast cancer, colon cancer, and head and neck cancer. Other cancers that can be treated according to the present invention include those characterized by elevated expression of one or both of PD-L1 and PD-L2 in a tissue sample to be tested.

[0103] "CDR" refers to one or more complementarity determining regions in the variable region of an immunoglobulin, typically defined using the Kabat numbering system.

[0104] "Chemotherapeutic agent" is a chemical compound that can be used to treat cancer. Classes of chemotherapeutic agents include, but are not limited to: alkylating agents, antimetabolites, kinase inhibitors, spindle poison plant alkaloids, cytotoxic / antitumor antibiotics, topoisomerase inhibitors, photosensitizers, antiestrogens and selective estrogen receptor modulators (SERMs), antiprogestins, estrogen receptor downregulators (ERDs), estrogen receptor antagonists, luteinizing hormone-releasing hormone agonists, antiandrogens, aromatase inhibitors, EGFR inhibitors, VEGF inhibitors, antisense oligonucleotides that inhibit gene expression involved in abnormal cell proliferation or tumor growth. Chemotherapeutic agents useful in the methods, compositions, and uses of the present invention include cell growth inhibitors and / or cytotoxic agents.

[0105] "Chimeric antibody" refers to an antibody in which a portion of the heavy and / or light chain is identical or homologous to the corresponding sequence of an antibody derived from a particular species (e.g., human) or belonging to a particular antibody class or subclass, while the remainder of the one or more chains is identical or homologous to the corresponding sequence of an antibody derived from another species (e.g., mouse) or belonging to another antibody class or subclass, and fragments of such antibodies, provided that they exhibit the desired biological activity.

[0106] Throughout the specification and claims, the term "comprising" or variations thereof, such as "comprise", "comprises", or "comprised of", is used in an inclusive sense, i.e., it specifies the presence of the stated property, but does not exclude the presence or addition of other properties that may materially enhance the operation or utility of any embodiment of the invention, unless the context requires otherwise due to the language of the expression or necessary implication.

[0107] "Conservative modified variants" or "conservative substitutions" refer to the replacement of an amino acid in a protein with another amino acid having similar characteristics (e.g., charge, side chain size, hydrophobicity / hydrophilicity, backbone conformation, and rigidity, etc.), such that changes can often be made without altering the biological activity or other desired properties of the protein, such as antigenic affinity and / or specificity. Those skilled in the art recognize that, in general, single amino acid substitutions in non-essential regions of a polypeptide do not substantially alter biological activity (see, e.g., Watson et al., (1987) Molecular Biology of the Gene, The Benjamin / Cummings Pub. Co., p. 224 (4th ed.)). Additionally, amino acid substitutions that are structurally or functionally similar are less likely to disrupt biological activity. Exemplary conservative substitutions are shown in Table 1.

[0108] Table 1: Exemplary conservative amino acid substitutions

[0109] Original Residue Conservative Substitution Ala(A) Gly;Ser Arg(R) Lys;His Asn(N) Gln;His Asp(D) Glu;Asn Cys(C) Ser;Ala Gln(Q) Asn Glu(E) Asp;Gln Gly(G) Ala His(H) Asn;Gln Ile(I) Leu;Val Leu(L) Ile;Val Lys(K) Arg;His Met(M) Leu;Ile;Tyr Phe(F) Tyr;Met;Leu Pro(P) Ala Ser(S) Thr Thr(T) Ser Trp(W) Tyr;Phe Tyr(Y) Trp;Phe Val(V) Ile;Leu

[0110] "Diagnostic anti-PD-L monoclonal antibody" refers to an mAb that specifically binds to the mature form of the designated PD-L (PD-L1 or PD-L2) expressed on the surface of certain mammalian cells. Mature PD-L lacks the pre-secretory leader sequence, also known as the leader peptide. The terms "PD-L" and "mature PD-L" are used interchangeably herein and, unless otherwise specified or obvious from the context, should be understood to refer to the same molecule.

[0111] As used herein, a diagnostic anti-human PD-L1 mAb or anti-hPD-L1 mAb refers to a monoclonal antibody that specifically binds to mature human PD-L1. The mature human PD-L1 molecule consists of amino acids 19 - 220 of the following sequence:

[0112] MRIFAVFIFMTYWHLLNAFTVTVPKDLYVVEYGSNMTIECKFPVEKQLDLAALIVYWEMEDKNIIQFVHGEEDLKVQHSSYRQRARLLKDQLSLGNAALQITDVKLQDAGVYRCMISYGGADYKRITVKVNAPYNKINQRILVVDPVTSEHELTCQAEGYPKAEVIWTSSDHQVLSGKTTTTNSKREEKLFNVTSTLRINTTTNEIFYCTFRRLDPEENHTAELVIPELPLAHPPNERTHLVILGAILLCLGVALTFIFRLRKGRMMDVKKCGIQDTNSKKQSDTHLEET(SEQ ID NO:21).

[0113] Specific examples of diagnostic anti-human PD-L1 mAbs that can be used as diagnostic mAbs for immunohistochemical (IHC) detection of PD-L1 expression in formalin-fixed paraffin-embedded (FFPE) tumor tissue sections are antibody 20C3 and antibody 22C3, which are described in WO 2014 / 100079. These antibodies contain the light and heavy chain variable region amino acid sequences shown in Table 2 below:

[0114]

[0115] Another anti-human PD-L1 mAb (Chen, B.J. et al., Clin Cancer Res 19:3462-3473 (2013)) reported to be useful for IHC detection of PD-L1 expression in FFPE tissue sections is a rabbit anti-human PD-L1 mAb publicly available from Sino Biological, Inc. (Beijing, P.R. China; catalog number 10084-R015).

[0116] As used herein, "framework region" or "FR" refers to the immunoglobulin variable region excluding the CDR regions.

[0117] "Human antibody" refers to an antibody that contains only human immunoglobulin protein sequences. If produced in a mouse, in mouse cells or in a hybridoma derived from mouse cells, a human antibody may contain mouse carbohydrate chains. Similarly, "mouse antibody" or "rat antibody" refers to an antibody that contains only mouse or rat immunoglobulin sequences, respectively.

[0118] "Humanized antibody" refers to an antibody containing sequences or antibody forms from non-human (e.g., murine) antibodies and human antibodies. Such antibodies contain minimal sequences derived from non-human immunoglobulins. Typically, the humanized antibody will contain substantially all of at least one and usually two variable domains, wherein all or substantially all of the hypervariable loops correspond to those of the non-human immunoglobulin and all or substantially all of the FR regions are those of human immunoglobulin sequences. The humanized antibody will optionally also contain an immunoglobulin constant region (Fc), typically at least a portion of the immunoglobulin constant region of a human immunoglobulin. When necessary, a prefix such as "hum", "hu" or "h" is added to the antibody clone name to distinguish the humanized antibody from the parental rodent antibody. Although certain amino acid substitutions may be included to increase affinity, increase the stability of the humanized antibody or for other reasons, the humanized form of the rodent antibody will generally contain the same CDR sequences as the parental rodent antibody.

[0119] "Hypervariable region" refers to the amino acid residues of an antibody that are responsible for antigen binding. The hypervariable region contains amino acid residues from "complementary determining regions" or "CDRs" (i.e., LC-CDR1, LC-CDR2 and LC-CDR3 in the light chain variable domain and HC-CDR1, HC-CDR2 and HC-CDR3 in the heavy chain variable domain). See Kabat et al., (1991) Sequences of Proteins of Immunological Interest, 5th ed., Public Health Service, National Institutes of Health, Bethesda, Md. (antibody CDR regions defined by sequence); see also Chothia and Lesk (1987) J. Mol. Biol. 196:901-917 (antibody CDR regions defined by structure). The term "framework" or "FR" residues refers to those variable domain residues other than the hypervariable region residues defined herein as CDR residues.

[0120] "Immunogenic agent" refers to a composition capable of inducing a humoral and / or cell-mediated immune response. Immunogenic agents can include, for example, attenuated cancer cells, tumor antigens, antigen-presenting cells (such as dendritic cells pulsed with tumor-derived antigens or nucleic acids), immunostimulatory cytokines (e.g., IL-2, IFNα2, GM-CSF) and cells transfected with genes encoding immunostimulatory cytokines (such as, but not limited to, GM-CSF).

[0121] "In need thereof" refers to in need of treatment.

[0122] As used herein, "Kabat" refers to the immunoglobulin alignment and numbering system pioneered by Elvin A. Kabat ((1991) Sequences of Proteins of Immunological Interest, 5th ed., Public Health Service, National Institutes of Health, Bethesda, Md.).

[0123] As used herein, "monoclonal antibody" or "mAb" or "Mab" refers to a population of antibodies that are substantially homologous, i.e., the amino acid sequences of the antibody molecules comprising the population are identical, except for possible naturally occurring mutations that may be present in minimal amounts. In contrast, conventional (polyclonal) antibody preparations typically comprise a variety of different antibodies that have different amino acid sequences in their variable domains, particularly their CDRs, which are usually specific for different epitopes. The modifier "monoclonal" indicates that the antibody is characterized as being obtained from a substantially homogeneous population of antibodies and should not be construed as requiring that the antibody be made by any particular method. For example, monoclonal antibodies used in accordance with the present invention may be prepared by the hybridoma method first described by Kohler et al., (1975) Nature 256:495, or may be prepared by recombinant DNA methods (see, e.g., U.S. Patent No. 4,816,567). "Monoclonal antibodies" may also be isolated from phage antibody libraries using, for example, the techniques described in Clackson et al., (1991) Nature 352:624-628 and Marks et al., (1991) J. Mol. Biol. 222:581-597. See also Presta (2005) J. Allergy Clin. Immunol. 116:731.

[0124] "Microsatellite instability (MSI)" refers to a form of genomic instability associated with defective DNA mismatch repair in tumors. See Boland et al., Cancer Research 58, 5258-5257, 1998. In one embodiment, MSI analysis can use five microsatellite markers recommended by the National Cancer Institute (NCI) of the United States: BAT25 (GenBank accession number 9834508), BAT26 (GenBank accession number 9834505), D5S346 (GenBank accession number 181171), D2S123 (GenBank accession number 187953), D17S250 (GenBank accession number 177030). Other markers can be used, for example, BAT40, BAT34C4, TGF-β-RII, and ACTC. Commercially available kits for MSI analysis include, for example, the Promega MSI multiplex PCR assay, the CDx (F1CDx) next-generation sequencing using DNA isolated from formalin-fixed paraffin-embedded (FFPE) tumor tissue samples based on in vitro diagnostic devices. CDx (F1CDx) next-generation sequencing.

[0125] "High-frequency microsatellite instability" or "microsatellite instability-high (MSI-H)" refers to a tumor in which two or more of the above five NCI markers display instability of their DNA or a total marker in their DNA ≥ 30-40% confirmed instability (i.e., having insertion / deletion mutations).

[0126] As used herein, "non-MSI-H cancer" refers to microsatellite stable (MSS) and low-frequency MSI (MSI-L) cancers.

[0127] "Microsatellite stable (MSS)" refers to a tumor in which none of the above-specified five NCI markers display instability of their DNA (i.e., having insertion / deletion mutations).

[0128] "Patient" (or "subject" or "individual" as referred to herein) refers to a mammal (e.g., rat, mouse, dog, cat, rabbit) that can be treated using the methods and compositions of the present invention, most preferably a human. In certain embodiments, the patient is an adult human patient. In other embodiments, the patient is a pediatric patient.

[0129] Unless otherwise defined, "PD-L1" or "PD-L2" expression refers to any detectable level of expression of the designated PD-L protein on the cell surface or of the designated PD-L mRNA within a cell or tissue. PD-L protein expression can be detected in an IHC assay of a tumor tissue section using a diagnostic PD-L antibody or by flow cytometry. Alternatively, PD-L protein expression on tumor cells can be detected by PET imaging using an agent (e.g., an antibody fragment, an affibody, etc.) that specifically binds to the desired PD-L target (e.g., PD-L1 or PD-L2). Techniques for detecting and measuring PD-L mRNA expression include reverse transcription polymerase chain reaction (RT-PCR) and real-time quantitative RT-PCR.

[0130] Several methods for quantifying PD-L1 protein expression in an IHC assay of a tumor tissue section have been described. See, e.g., Thompson et al., PNAS 101(49):17174-17179 (2004); Thompson et al., Cancer Res. 66:3381-3385 (2006); Gadiot et al., Cancer 117:2192-2201 (2011); Taube et al., Sci Transl Med 4, 127ra37 (2012); and Toplian et al., New Eng. J Med. 366(26):2443-2454 (2012).

[0131] One method employs a simple binary endpoint of positive or negative for PD-L1 expression, where a positive result is defined by the percentage of tumor cells showing histological evidence of cell surface membrane staining. A tumor tissue section is considered positive for PD-L1 expression if at least 1% and preferably 5% of the total tumor cells show histological evidence of cell surface membrane staining.

[0132] In another method, PD-L1 expression in tumor tissue sections is quantified in tumor cells as well as in infiltrating immune cells consisting predominantly of lymphocytes. The percentages of tumor cells and infiltrating immune cells showing membrane staining are quantified separately as <5%, 5 to 9%, and then increasing in 10% increments up to 100%. For tumor cells, if the score is <5%, the PD-L1 expression is considered negative, and if the score ≥5%, it is positive. The PD-L1 expression in the immune infiltration is reported as a semi-quantitative measurement called the adjusted inflammation score (AIS), which is determined by multiplying the percentage of membrane-stained cells by the infiltration intensity, with the infiltration intensity classified as none (0), mild (1 point, rare lymphocytes), moderate (2 points, focal infiltration of lymphohistiocytic aggregates into the tumor), or severe (3 points, diffuse infiltration). If the AIS ≥5, the tumor tissue section is counted as PD-L1 expression positive by immune infiltration. By using a scoring method to evaluate PD-L1 expression in tumor cells and infiltrating immune cells in tissue sections, PD-L1 protein expression scoring can also be performed on tumor tissue sections stained by IHC with a diagnostic PD-L1 antibody. See WO 2014 / 165422. A PD-L1 scoring method includes examining each tumor nest in a tissue section for staining and assigning to the tissue section one or both of a modified H score (MHS) and a modified proportion score (MPS). To assign the MHS, in all examined tumor nests, four separate percentages are estimated for all viable tumor cells and stained mononuclear inflammatory cells: (a) cells with no staining (intensity = 0), (b) weakly stained (intensity = 1+), (c) moderately stained (intensity = 2+), and (d) strongly stained (intensity = 3+). Cells must have at least partial membrane staining to be included in the weakly, moderately, or strongly stained percentages. The estimated percentages (which sum to 100%) are then entered into the formula 1x(percentage of weakly stained cells) + 2x(percentage of moderately stained cells) + 3x(percentage of strongly stained cells), and the result is assigned as the MHS to the tissue section. The MPS is assigned by estimating the percentage of cells with at least any intensity of partial membrane staining among all viable tumor cells and stained mononuclear inflammatory cells in all examined tumor nests, and the resulting percentage is assigned as the MPS to the tissue section. In a particular embodiment, if the MHS or MPS is positive, the tumor is designated as positive for PD-L1 expression.

[0133] Another method for scoring / quantifying PD-L1 expression in tumors is the "Combined Positive Score" or "CPS", which refers to an algorithm for determining the PD-L1 expression score from a patient's tumor sample. CPS can be used to select patients for treatment with specific treatment regimens, including treatment methods involving administration of anti-PD-1 antibodies, where the expression of PD-L1 in a specific patient population is associated with a higher response rate relative to the same patient population that does not express PD-L1. CPS is determined by determining the number of viable PD-L1-positive tumor cells, the number of viable PD-L-1-negative tumor cells, and the number of viable PD-L1-positive mononuclear inflammatory cells (MICs) in the tumor tissue of a patient with a tumor, and calculating the CPS using the following formula:

[0134]

[0135] In a particular embodiment, the PD-L1 expression scoring method used is the "Lymphocyte Proportion Score". Lymphomas are characterized by homogeneous populations of fused cells that obliterate the architecture of lymph nodes or metastatic sites. "LPS" or "Lymphocyte Proportion Score" is the percentage of the population of cells expressing PD-L1. When determining LPS, no attempt is made to distinguish between true tumor cells and reactive cells. PD-L1 expression is characterized by partial or complete membrane staining of any intensity.

[0136] Another scoring method for PD-L1 expression is the "TPS" or "Tumor Proportion Score", which is the percentage of tumor cells expressing PD-L1 on the cell membrane. TPS generally includes the percentage of tumor cells expressing PD-L1 at any intensity (weak, moderate, or strong), which can be determined by immunohistochemical assays using diagnostic anti-human PD-L1 mAbs (e.g., antibodies 20C3 and 22C3 as described above). If there is membrane staining, including cells with partial membrane staining, the cells are considered to express PD-L1.

[0137] The level of PD-L mRNA expression can be compared to the mRNA expression level of one or more reference genes (such as ubiquitin C) commonly used in quantitative RT-PCR.

[0138] In certain embodiments, the PD-L1 expression level of malignant cells and / or tumor-infiltrating immune cells is determined to be "overexpressed" or "elevated" based on a comparison with the PD-L1 expression level (protein and / or mRNA) of an appropriate control. For example, the control PD-L1 protein or mRNA expression level can be the level quantified in non-malignant cells of the same type or in matched normal tissue sections. In certain embodiments, PD-L1 expression is determined to be elevated in a tumor sample if the PD-L1 protein (and / or PD-L1 mRNA) in the sample is at least 10%, 20%, or 30% higher than in the control.

[0139] "Pembrolizumab" (previously known as MK-3475, SCH 900475, and lambrolizumab), or "pembro" as referred to herein, is a humanized IgG4 mAb having the structure described in WHO Drug Information, Vol. 27, No. 2, pages 161-162 (2013), which includes the heavy and light chain amino acid sequences and CDRs described in Table 3. Pembrolizumab has been approved by the US FDA as described in the TM prescribing information for KEYTRUDA (Merck & Co., Inc., Whitehouse Station, NJ USA; initial US approval 2014, updated March 2021).

[0140] As used herein, "pembrolizumab variant" refers to a monoclonal antibody that contains the same heavy and light chain sequences as those in pembrolizumab, except for having three, two, or one conservative amino acid substitutions at positions outside the light chain CDRs and six, five, four, three, two, or one conservative amino acid substitutions at positions outside the heavy chain CDRs. For example, the variant positions are in the FR regions or the constant regions, and optionally have a deletion of the C-terminal lysine or glycine residue of the heavy chain. In other words, pembrolizumab and pembrolizumab variants contain the same CDR sequences but differ from each other by having no more than three or six other conservative amino acid substitutions in their full-length light and heavy chain sequences, respectively. Pembrolizumab variants are substantially identical to pembrolizumab with respect to the following properties: binding affinity for PD-1 and the ability to block the binding of each of PD-L1 and PD-L2 to PD-1.

[0141] "Pharmaceutical formulation" or "pharmaceutical composition" refers to a formulation in a form that permits the active ingredient to be effective and that does not contain other ingredients that are toxic to the subject to which the formulation is to be administered.

[0142] "Pharmaceutically acceptable" refers to excipients (carriers, additives) and compositions that can be reasonably administered to a subject to provide an effective dose of the active ingredient used and that are "generally regarded as safe", e.g., when administered to humans, they are physiologically tolerable and generally do not produce allergic or similar adverse reactions such as stomach discomfort. In another embodiment, the term refers to molecular entities and compositions that are approved by a federal or state government regulatory agency or listed in the United States Pharmacopeia or other recognized pharmacopeias for use in animals (more particularly humans).

[0143] Pharmacokinetic "steady state" refers to a period of time during which the drug concentration accumulates to a maximum due to multiple administrations and the systemic drug exposure is considered uniform after each subsequent administration; in the specific case of pembrolizumab, steady state is reached at approximately 16 weeks of administration and after approximately 16 weeks of administration.

[0144] "Platinum-containing chemotherapy" (also referred to as platinum agents) refers to the use of one or more chemotherapeutic agents for the treatment of cancer, where the chemotherapeutic agents are platinum coordination complexes. Platinum-containing chemotherapeutic agents are alkylating agents that crosslink DNA, resulting in ineffective DNA mismatch repair and typically leading to apoptosis. Examples of platinum agents include cisplatin, carboplatin, and oxaliplatin.

[0145] As used herein, "RECIST 1.1 response criteria" refers to the definitions set forth in Eisenhauer, E.A. et al., Eur. J. Cancer 45:228-247 (2009) for target or non-target lesions, which depend on the context of the measured response.

[0146] "Therapeutic agent" refers to other agents relative to an anti-PD-1 antibody or its antigen-binding fragment. Therapeutic agents can be, for example, chemotherapeutic agents, biotherapeutic agents, or immunogenic agents.

[0147] "Tissue section" refers to a single part or slice of a tissue sample, e.g., a thin section of tissue cut from a sample of normal tissue or a tumor.

[0148] "Tumor", as applied to a subject diagnosed with or suspected of having cancer, refers to any sized malignant or potentially malignant neoplasm or mass of tissue, including primary tumors and secondary neoplasms. A solid tumor is an abnormal growth or mass of tissue that generally does not contain cysts or fluid areas. Different types of solid tumors are named for the cell types that form them. Examples of solid tumors are sarcomas, carcinomas, and lymphomas. Leukemias (blood cancers) generally do not form solid tumors (National Cancer Institute, Dictionary of Cancer Terms).

[0149] As used herein, "tumor mutational burden" or "TMB" refers to the number of somatic mutations in the tumor genome and / or the number of somatic variants per region of the tumor genome. High TMB (or TMB-H) refers to a tumor with a high mutational burden. In certain embodiments, a tumor is referred to as TMB-H if it contains ≥10 mutations / megabase (Mut / Mb). Tests approved by the US FDA, such as CDx, can be used for solid tumors to determine whether the solid tumor is TMB-H (i.e., has ≥10 mutations / megabase).

[0150] As used herein, "variable region" or "V region" refers to the segment of the IgG chain that is variable in sequence between different antibodies. It extends to Kabat residue 109 in the light chain and to 113 in the heavy chain.

[0151] "PH 20" refers to the wild-type PH20 hyaluronidase of SEQ ID NO:16.

[0152] As used herein, "PH20 variant" refers to a variant of PH20 having an amino acid residue substitution (including M345T, S347T, M348K, K349E, L352Q, L353A, L354I, D355K, N356E, E359D, and I361T) in SEQ ID NO:16.

[0153] "PH20 variant fragment" or "its PH20 variant fragment" or "fragment of a PH20 variant" is a PH20 variant having an N-terminal deletion of amino acid residues 1-36, 1-37, 1-38, 1-39, 1-40, 1-41 or 1-42 of SEQ ID NO:16; and / or a C-terminal deletion of amino acid residues 455-509, 456-509, 457-509, 458-509, 459-509, 460-509, 461-509, 462-509, 463-509, 464-509, 465-509, 466-509, 467-509, 468-509, 469-509, 470-509, 471-509, 472-509, 473-509, 474-509, 475-509, 476-509, 477-509, 478-509, 479-509, 480-509, 481-509, 482-509, 483-509, 484-509, 485-509, 486-509, 487-509, 488-509, 489-509, 490-509, 491-509, 492-509, 493-509, 494-509, 495-509, 496-509, 497-509, 498-509, 499-509, 500-509, 501-509, 502-509, 503-509, 504-509, 505-509, 506-509, 507-509, 508-509 or 509, wherein numbering is referenced to SEQ ID NO:16.

[0154] "Unit" or "U" refers to a unit of hyaluronidase activity: the amount of a PH20 variant or its fragment that causes a change in optical intensity at 600 nm under conditions suitable for hyaluronic acid and enzyme reactions and is calculated using an activity standard according to a calibration curve. Examples of the assay are described in Example 4 of US 2022 / 0089738. Hyaluronic acid (HA) binds to albumin and the albumin-HA complex produces turbidity. When HA is hydrolyzed by hyaluronidase, the turbidity of the albumin-HA complex decreases. Thus, this measurement measures turbidity to determine the hyaluronidase activity of a PH20 variant or its fragment. Hyaluronidase activity is based on the following reaction:

[0155] Hyaluronic acid ––––––––––––> Disaccharides and monosaccharides + Smaller hyaluronic acid fragments. Those skilled in the art understand that hyaluronidase activity in units / mg of hyaluronidase can vary depending on the purity of the hyaluronidase, the method of preparation, etc.

[0156] II. PD-1 Antibodies and Antigen-Binding Fragments Useful in the Invention

[0157] Examples of mAbs that bind to human PD-1 and can be used in the formulations, treatment methods, compositions, and uses of the present invention are described in US 7,521,051, US 8,008,449, and US 8,354,509. Specific anti-human PD-1 mAbs that can be used as PD-1 antagonists or anti-PD-1 antibodies in the treatment methods, compositions, and uses of the present invention include: pembrolizumab (previously known as MK-3475, SCH 900475, and lambrolizumab), and a humanized IgG4 mAb having the structure described in WHO Drug Information, Vol. 27, No. 2, pp. 161-162 (2013).

[0158] In certain embodiments of the treatment methods, compositions, kits, and uses of the present invention, the anti-PD-1 antibody or antigen-binding fragment thereof comprises: (a) light chain CDRs LC-CDR1, LC-CDR2, and LC-CDR3, which comprise the amino acid sequences shown in SEQ ID NOs: 1, 2, and 3, respectively, and heavy chain CDRs HC-CDR1, HC-CDR2, and HC-CDR3, which comprise the amino acid sequences shown in SEQ ID NOs: 6, 7, and 8, respectively. In other embodiments, the anti-PD-1 antibody or antigen-binding fragment thereof is a humanized antibody. In other embodiments, the anti-PD-1 antibody or antigen-binding fragment thereof is a chimeric antibody. In certain embodiments, the anti-PD-1 antibody or antigen-binding fragment thereof is a monoclonal antibody.

[0159] In other embodiments of the treatment methods, compositions, kits, and uses of the present invention, the anti-PD-1 antibody or antigen-binding fragment thereof specifically binds to human PD-1 and comprises (a) a heavy chain variable region that comprises the amino acid sequence shown in SEQ ID NO: 9 or a variant of SEQ ID NO: 9, and (b) a light chain variable region that comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 4 or a variant of SEQ ID NO: 4. In one embodiment, the anti-PD-1 antibody or antigen-binding fragment thereof that specifically binds to human PD-1 comprises (a) a heavy chain variable region that comprises the amino acid sequence shown in SEQ ID NO: 9, and (b) a light chain variable region that comprises the amino acid sequence of SEQ ID NO: 4.

[0160] A variant of a heavy chain variable region sequence or a full-length heavy chain sequence is identical to the reference sequence, except that it has at most 17 conservative amino acid substitutions in the framework regions (i.e., outside the CDRs), and preferably has less than ten, nine, eight, seven, six, or five conservative amino acid substitutions in the framework regions. A variant of a light chain variable region sequence or a full-length light chain sequence is identical to the reference sequence, except that it has at most five conservative amino acid substitutions in the framework regions (i.e., outside the CDRs), and preferably has less than four, three, or two conservative amino acid substitutions in the framework regions.

[0161] In another embodiment of the therapeutic methods, compositions, kits, and uses of the present invention, the anti-PD-1 antibody or its antigen-binding fragment is a monoclonal antibody that specifically binds to human PD-1 and comprises (a) a heavy chain that comprises or consists of an amino acid sequence as shown in any one of SEQ ID NOs: 10-15 or a variant thereof; and (b) a light chain that comprises or consists of an amino acid sequence as shown in SEQ ID NO: 5 or a variant thereof. In one embodiment, the anti-PD-1 antibody or its antigen-binding fragment is a monoclonal antibody that specifically binds to human PD-1 and comprises (a) a heavy chain that consists of an amino acid sequence as shown in any one of SEQ ID NOs: 10-15; and (b) a light chain that consists of an amino acid sequence as shown in SEQ ID NO: 5.

[0162] In yet another embodiment of the therapeutic methods, compositions, kits, and uses of the present invention, the anti-PD-1 antibody or its antigen-binding fragment is a monoclonal antibody that specifically binds to human PD-1 and comprises (a) a heavy chain that comprises or consists of the amino acid sequence as shown in SEQ ID NO: 11; and (b) a light chain that comprises or consists of the amino acid sequence as shown in SEQ ID NO: 5.

[0163] Table 3 below provides a list of the amino acid sequences of exemplary anti-PD-1 mAbs for use in the therapeutic methods, compositions, kits, and uses of the present invention.

[0164] Table 3: Exemplary anti-PD-1 antibody sequences

[0165]

[0166]

[0167]

[0168] As is well known to those skilled in the art, pyroglutamic acid is the conjugate acid of pyroglutamate and is in equilibrium with pyroglutamate in solution.

[0169] III. Human hyaluronidase

[0170] Six forms of hyaluronidase are known to exist in humans: Hyal1, Hyal2, Hyal3, Hyal4, HyalPS1, and PH20 / SPAM1. Recombinant forms of these hyaluronidases with modifications, mutations, additions, or truncations can be used in the disclosed methods, uses, compositions, and kits. See, for example, U.S. Patent Nos. 7,767,429, 8,431,380, 7,871,607, International Publication No. WO 2020 / 022791, U.S. Patent Publication No. US2006 / 0104968, and European Patent 1858926, as well as in many other patents and publications. Examples of such agents are the known agents PEGPH20 or rHuPH20. The methods, uses, compositions, and kits of the present invention encompass the use of any human hyaluronidase or fragment thereof, or variant thereof or fragment thereof.

[0171] PH20 Variants and Their Fragments

[0172] In one embodiment, the PH20 variant or fragment thereof further comprises an amino acid residue substitution at one or more positions selected from: T341, L342, S343, I344, and N363. In one embodiment, the PH20 variant or fragment thereof further comprises one or more amino acid residue substitutions selected from: T341A, T341C, T341D, T341G, T341S, L342W, S343E, I344N, and N363G.

[0173] In one embodiment of the PH20 variant or fragment thereof, the amino acid residue substitution is selected from the group consisting of the following amino acid residue substitution groups:

[0174] (a) T341S, L342W, S343E, I344N, M345T, S347T, M348K, K349E, L352Q, L353A, L354I, D355K, N356E, E359D, and I361T;

[0175] (b) L342W, S343E, I344N, M345T, S347T, M348K, K349E, L352Q, L353A, L354I, D355K, N356E, E359D, and I361T;

[0176] (c) M345T, S347T, M348K, K349E, L352Q, L353A, L354I, D355K, N356E, E359D, I361T, and N363G;

[0177] (d) T341G, L342W, S343E, I344N, M345T, S347T, M348K, K349E, L352Q, L353A, L354I, D355K, N356E, E359D, and I361T;

[0178] (e) T341A, L342W, S343E, I344N, M345T, S347T, M348K, K349E, L352Q, L353A, L354I, D355K, N356E, E359D, and I361T;

[0179] (f) T341C, L342W, S343E, I344N, M345T, S347T, M348K, K349E, L352Q, L353A, L354I, D355K, N356E, E359D, and I361T;

[0180] (g) T341D, L342W, S343E, I344N, M345T, S347T, M348K, K349E, L352Q, L353A, L354I, D355K, N356E, E359D, and I361T;

[0181] (h) I344N, M345T, S347T, M348K, K349E, L352Q, L353A, L354I, D355K, N356E, E359D, and I361T; and

[0182] (i) S343E, I344N, M345T, S347T, M348K, K349E, L352Q, L353A, L354I, D355K, N356E, E359D, and I361T.

[0183] In one embodiment of the PH20 variant or a fragment thereof, the amino acid residue substitutions consist of: T341S, L342W, S343E, I344N, M345T, S347T, M348K, K349E, L352Q, L353A, L354I, D355K, N356E, E359D, and I361T.

[0184] In one aspect of the foregoing embodiments of the PH20 variant fragment, the PH20 variant fragment has an N-terminal deletion comprising amino acid residues 1-36, 1-37, 1-38, 1-39, 1-40, 1-41 or 1-42 in SEQ ID NO: 16. In another embodiment, the PH20 variant has an N-terminal deletion comprising amino acid residues 1-36 in SEQ ID NO: 16. In another embodiment, the PH20 variant has an N-terminal deletion of amino acid residues 1-37 of SEQ ID NO: 16. In another embodiment, the PH20 variant has an N-terminal deletion of amino acid residues 1-38 of SEQ ID NO: 16.

[0185] In another aspect of the foregoing embodiments of the PH20 variant fragment, the PH20 variant fragment has a C-terminal deletion of one or more of amino acid residues 455-509, 456-509, 457-509, 458-509, 459-509, 460-509, 461-509, 462-509, 463-509, 464-509, 465-509, 466-509, 467-509, 468-509, 469-509, 470-509, 471-509, 472-509, 473-509, 474-509, 475-509, 476-509, 477-509, 478-509, 479-509, 480-509, 481-509, 482-509, 483-509, 484-509, 485-509, 486-509, 487-509, 488-509, 489-509, 490-509, 491-509, 492-509, 493-509, 494-509, 495-509, 496-509, 497-509, 498-509, 499-509, 500-509, 501-509, 502-509, 503-509, 504-509, 505-509, 506-509, 507-509, 508-509 or 509, wherein the numbering refers to SEQ ID NO:16. In one embodiment, the PH20 variant fragment thereof has a C-terminal deletion of amino acid residues 455-509, 458-509, 461-509, 464-509, 465-509, 466-509, 467-509, 468-509, 470-509, 471-509, 472-509, 473-509, 474-509, 475-509, 476-509, 478-509, 480-509, 482-509, 484-509, 486-509, 488-509 or 490-509, wherein the numbering refers to SEQ ID NO:16. In one embodiment, the PH20 variant fragment has a C-terminal deletion of amino acid residues 468-509, wherein the numbering refers to SEQ ID NO:16.

[0186] In one embodiment, the PH20 variant fragment consists of the amino acid sequence shown in SEQ ID NO:17 or 18. In other embodiments, the PH20 variant or a fragment thereof is any sequence disclosed in Table 11 of EP3636752.

[0187] Table 4: Hyaluronidase and Exemplary Variants

[0188]

[0189] In one aspect of the methods, compositions, kits, and uses of the present invention, the pharmaceutical composition comprises about 165 mg / mL of an anti-human PD-1 antibody or an antigen-binding fragment thereof, about 10 mM histidine buffer, about 10 mM L-methionine or a pharmaceutically acceptable salt thereof, about 7% w / v sucrose, about 0.02% w / v polysorbate 80, and a PH20 variant or fragment. In one embodiment, the pharmaceutical composition comprises about 130 mg / mL of an anti-human PD-1 antibody or an antigen-binding fragment thereof, about 10 mM histidine buffer, about 10 mM L-methionine or a pharmaceutically acceptable salt thereof, about 7% w / v sucrose, about 0.02% w / v polysorbate 80, and a PH20 variant or fragment.

[0190] In certain embodiments of the pharmaceutical composition, the PH20 variant or a fragment thereof is present at a concentration of about 1000 U / ml. In another embodiment, the concentration of the PH20 variant or a fragment thereof is about 1500 U / ml. In another embodiment, the concentration of the PH20 variant or a fragment thereof is about 2000 U / ml. In another embodiment, the concentration of the PH20 variant or a fragment thereof is about 3000 U / ml. In another embodiment, the concentration of the PH20 variant or a fragment thereof is about 4000 U / ml. In another embodiment, the concentration of the PH20 variant or a fragment thereof is about 5000 U / ml. In another embodiment, the concentration of the PH20 variant or a fragment thereof is about 6000 U / ml. In a further embodiment, the concentration of the PH20 variant or a fragment thereof is about 1000 - 6000 U / ml. In a further embodiment, the concentration of the PH20 variant or a fragment thereof is about 2000 - 5000 U / ml.

[0191] In certain embodiments of the pharmaceutical composition, the PH20 variant or a fragment thereof is present at a concentration of about 150 U / ml. In another embodiment, the concentration of the PH20 variant or a fragment thereof is about 300 U / ml. In another embodiment, the concentration of the PH20 variant or a fragment thereof is about 600 U / ml. In another embodiment, the concentration of the PH20 variant or a fragment thereof is about 750 U / ml. In a further embodiment, the concentration of the PH20 variant or a fragment thereof is about 150 - 5000 U / ml. In a further embodiment, the concentration of the PH20 variant or a fragment thereof is about 500 - 8000 U / ml.

[0192] In another aspect of the methods, compositions, kits, and uses of the present invention, the pharmaceutical composition comprises about 165 mg / mL anti-human PD-1 antibody and 2000 U / ml PH20 variant or fragment. In one embodiment, the pharmaceutical composition comprises about 130 mg / mL anti-human PD-1 antibody and 2000 U / ml PH20 variant or fragment.

[0193] rHuPH20 and Its Fragments

[0194] rHuPH20, also known as Composed of the amino acid sequence in SEQ ID NO:20, which is amino acid residues 36 - 482 of wild - type human PH20 in SEQ ID NO:16 (amino acid residues 1 - 36 are the signal peptide sequence). In one embodiment, rHuPH20 or a variant or fragment is amino acid residues 36 - 464, 36 - 465, 36 - 466, 36 - 467, 36 - 468, 36 - 469, 36 - 470, 36 - 471, 36 - 472, 36 - 473, 36 - 474, 36 - 475, 36 - 476, 36 - 477, 36 - 478, 36 - 479, 36 - 480, 36 - 481, 36 - 482 or 36 - 483 of SEQ ID NO:16; amino acid residues 37 - 464, 37 - 465, 37 - 466, 37 - 467, 37 - 468, 37 - 469, 37 - 470, 37 - 471, 37 - 472, 37 - 473, 37 - 474, 37 - 475, 37 - 476, 37 - 477, 37 - 478, 37 - 479, 37 - 480, 37 - 481, 37 - 482 or 37 - 483 of SEQ ID NO:16; amino acid residues 38 - 464, 38 - 465, 38 - 466, 38 - 467, 38 - 468, 38 - 469, 38 - 470, 38 - 471, 38 - 472, 38 - 473, 38 - 474, 38 - 475, 38 - 476, 38 - 477, 38 - 478, 38 - 479, 38 - 480, 38 - 481, 38 - 482 or 38 - 483 of SEQ ID NO:16; amino acid residues 39 - 464, 39 - 465, 39 - 466, 39 - 467, 39 - 468, 39 - 469, 39 - 470, 39 - 471, 39 - 472, 39 - 473, 39 - 474, 39 - 475, 39 - 476, 39 - 477, 39 - 478, 39 - 479, 39 - 480, 39 - 481, 39 - 482 or 39 - 483 of SEQ ID NO:16; amino acid residues 40 - 464, 40 - 465, 40 - 466, 40 - 467, 40 - 468, 40 - 469, 40 - 470, 40 - 471, 40 - 472, 40 - 473, 40 - 474, 40 - 475, 40 - 476, 40 - 477, 40 - 478, 40 - 479, 40 - 480, 40 - 481, 340 - 482 or 40 - 483 of SEQ ID NO:16;Amino acid residues 41 - 464, 41 - 465, 41 - 466, 41 - 467, 41 - 468, 41 - 469, 41 - 470, 41 - 471, 41 - 472, 41 - 473, 41 - 474, 41 - 475, 41 - 476, 41 - 477, 41 - 478, 41 - 479, 41 - 480, 41 - 481, 41 - 482 or 41 - 48 of SEQ ID NO:16; amino acid residues 42 - 464, 42 - 465, 42 - 466, 42 - 467, 42 - 468, 42 - 469, 42 - 470, 42 - 471, 42 - 472, 42 - 473, 42 - 474, 42 - 475, 42 - 476, 42 - 477, 42 - 478, 42 - 479, 42 - 480, 42 - 481, 42 - 482 or 42 - 483 of SEQ ID NO:16. In a preferred embodiment, the rHuPH20 variant consists of amino acid residues 36 - 483 of SEQ ID NO:16 (which is SEQ ID NO:19). In a preferred embodiment, the rHuPH20 fragment consists of amino acid residues 36 - 477 of SEQ ID NO:16. In a preferred embodiment, the rHuPH20 fragment consists of amino acid residues 36 - 478 of SEQ ID NO:16. In a preferred embodiment, the rHuPH20 fragment consists of amino acid residues 36 - 479 of SEQ ID NO:16. In a preferred embodiment, the rHuPH20 fragment consists of amino acid residues 36 - 480 of SEQ ID NO:16. In a preferred embodiment, the rHuPH20 fragment consists of amino acid residues 36 - 481 of SEQ ID NO:16. In a further embodiment, the rHuPH20 variant or fragment is those disclosed in U.S. Patent No. 7,767,429, the entire content of which is incorporated herein by reference.;

[0195] Table 5: Hyaluronidase and Exemplary Variants

[0196]

[0197] In one aspect, the methods, compositions, kits, and uses of the present invention utilize a pharmaceutical composition comprising about 165 mg / mL anti-human PD-1 antibody, about 10 mM histidine residue, about 10 mM L-methionine or a pharmaceutically acceptable salt thereof, about 7% w / v sucrose, about 0.02% w / v polysorbate 80, and rHuPH20 or a variant or fragment thereof. In one embodiment, the pharmaceutical composition comprises about 130 mg / mL anti-human PD-1 antibody or an antigen-binding fragment thereof, about 10 mM histidine buffer, about 10 mM L-methionine or a pharmaceutically acceptable salt thereof, about 7% w / v sucrose, about 0.02% w / v polysorbate 80, and rHuPH20 or a variant or fragment thereof.

[0198] In certain embodiments of the pharmaceutical composition, rHuPH20 or a variant or fragment thereof is present at a concentration of about 1000 U / ml. In another embodiment, the concentration of rHuPH20 or a variant or fragment thereof is about 1500 U / ml. In another embodiment, the concentration of rHuPH20 or a variant or fragment thereof is about 2000 U / ml. In another embodiment, the concentration of rHuPH20 or a variant or fragment thereof is about 3000 U / ml. In another embodiment, the concentration of rHuPH20 or a variant or fragment thereof is about 4000 U / ml. In another embodiment, the concentration of rHuPH20 or a variant or fragment thereof is about 5000 U / ml. In another embodiment, the concentration of rHuPH20 or a variant or fragment thereof is about 6000 U / ml. In a further embodiment, the concentration of rHuPH20 or a variant or fragment thereof is about 1000 - 6000 U / ml. In a further embodiment, the concentration of rHuPH20 or a variant or fragment thereof is about 2000 - 5000 U / ml.

[0199] In certain embodiments of the pharmaceutical composition, rHuPH20 or a variant or fragment thereof is present at a concentration of about 150 U / ml. In another embodiment, the concentration of rHuPH20 or a variant or fragment thereof is about 300 U / ml. In another embodiment, the concentration of rHuPH20 or a variant or fragment thereof is about 600 U / ml. In another embodiment, the concentration of rHuPH20 or a variant or fragment thereof is about 750 U / ml. In a further embodiment, the concentration of rHuPH20 or a variant or fragment thereof is about 150 - 5000 U / ml. In a further embodiment, the concentration of rHuPH20 or a variant or fragment thereof is about 500 to 8000 U / ml.

[0200] In another aspect, the methods, compositions, kits and uses of the present invention utilize a pharmaceutical composition comprising about 165 mg / mL anti-human PD-1 antibody or antigen-binding fragment thereof and about 2000 U / ml rHuPH20 or variant or fragment. In one embodiment, the pharmaceutical composition comprises about 130 mg / mL anti-human PD-1 antibody or antigen-binding fragment thereof and about 2000 U / ml rHuPH20 or variant or fragment.

[0201] IV. Methods and Uses of the Present Invention

[0202] The present invention provides a method for treating cancer in a human patient in need thereof, which comprises subcutaneously administering to the patient a dose of about 600 mg to about 1000 mg of an anti-PD-1 antibody or antigen-binding fragment thereof and human hyaluronidase approximately every six weeks. In another aspect, the present invention provides the use of an anti-PD-1 antibody or antigen-binding fragment thereof and human hyaluronidase for the preparation of a medicament for treating cancer in a human patient, wherein a dose of about 600 mg to about 1000 mg of an anti-PD-1 antibody or antigen-binding fragment thereof is subcutaneously administered to the patient approximately every six weeks. In another aspect, the present invention provides the use of an anti-PD-1 antibody or antigen-binding fragment thereof for the preparation of a medicament for treating cancer in a human patient, wherein a dose of about 600 mg to about 1000 mg of an anti-PD-1 antibody or antigen-binding fragment thereof and human hyaluronidase are co-subcutaneously administered to the patient approximately every six weeks. In another aspect, the present invention provides a pharmaceutical composition for treating cancer in a human patient, which comprises an anti-PD-1 antibody or antigen-binding fragment thereof and human hyaluronidase, wherein a dose of about 600 mg to about 1000 mg of an anti-PD-1 antibody or antigen-binding fragment thereof and human hyaluronidase are subcutaneously administered to the patient approximately every six weeks. In another aspect, the present invention provides a pharmaceutical composition for treating cancer in a human patient, which comprises an anti-PD-1 antibody or antigen-binding fragment thereof, wherein a dose of about 600 mg to about 1000 mg of an anti-PD-1 antibody or antigen-binding fragment thereof and human hyaluronidase are co-subcutaneously administered to the patient approximately every six weeks.

[0203] The present invention also provides a method for treating cancer in a human patient in need thereof, which comprises subcutaneously administering to the patient a dose of about 600 mg to about 1000 mg of an anti-PD-1 antibody or an antigen-binding fragment thereof approximately every six weeks. In another aspect, the present invention provides the use of an anti-PD-1 antibody or an antigen-binding fragment thereof for the preparation of a medicament for treating cancer in a human patient, wherein a dose of about 600 mg to about 1000 mg of the anti-PD-1 antibody or an antigen-binding fragment thereof is subcutaneously administered to the patient approximately every six weeks. In another aspect, the present invention provides an anti-PD-1 antibody or an antigen-binding fragment thereof for use in treating cancer in a human patient, wherein a dose of about 600 mg to about 1000 mg of the anti-PD-1 antibody or an antigen-binding fragment thereof is subcutaneously administered to the patient approximately every six weeks.

[0204] The present invention further provides a method for treating cancer in a human patient in need thereof, which comprises subcutaneously administering to the patient about 300 mg to about 500 mg of an anti-PD-1 antibody or an antigen-binding fragment thereof and human hyaluronidase approximately every three weeks. In another aspect, the present invention provides the use of an anti-PD-1 antibody or an antigen-binding fragment thereof and human hyaluronidase for the preparation of a medicament for treating cancer in a human patient, wherein a dose of about 300 mg to about 500 mg of the anti-PD-1 antibody or an antigen-binding fragment thereof and human hyaluronidase is subcutaneously administered to the patient approximately every three weeks. In another aspect, the present invention provides the use of an anti-PD-1 antibody or an antigen-binding fragment thereof for the preparation of a medicament for treating cancer in a human patient, wherein a dose of about 300 mg to about 500 mg of the anti-PD-1 antibody or an antigen-binding fragment thereof and human hyaluronidase is co-subcutaneously administered to the patient approximately every three weeks. In another aspect, the present invention provides a pharmaceutical composition for treating cancer in a human patient, which comprises an anti-PD-1 antibody or an antigen-binding fragment thereof and human hyaluronidase, wherein a dose of about 300 mg to about 500 mg of the anti-PD-1 antibody or an antigen-binding fragment thereof and human hyaluronidase is subcutaneously administered to the patient approximately every three weeks. In another aspect, the present invention provides a pharmaceutical composition for treating cancer in a human patient, which comprises an anti-PD-1 antibody or an antigen-binding fragment thereof, wherein a dose of about 300 mg to 500 mg of the anti-PD-1 antibody or an antigen-binding fragment thereof and human hyaluronidase is co-administered subcutaneously to the patient.

[0205] In a particular embodiment of the present invention, the anti-PD-1 antibody or an antigen-binding fragment thereof is pembrolizumab. In other embodiments, the anti-PD-1 antibody or an antigen-binding fragment thereof is a pembrolizumab variant.

[0206] In an embodiment of any of the methods or uses of the present invention, the bioavailability of the combination of an anti-PD-1 antibody or an antigen-binding fragment thereof and human hyaluronidase is 55-60%. In an embodiment of any of the methods or uses described herein, the bioavailability of the combination of an anti-PD-1 antibody or an antigen-binding fragment thereof and human hyaluronidase is 57-59%. In an embodiment of any of the methods or uses described herein, the bioavailability of the combination of an anti-PD-1 antibody or an antigen-binding fragment thereof and human hyaluronidase is 57%.

[0207] In an embodiment of any of the methods or uses of the present invention, subcutaneous administration of an anti-PD-1 antibody or an antigen-binding fragment thereof results in the C of the antibody or the antigen-binding fragment trough of the anti-PD-1 antibody or an antigen-binding fragment thereof at a dose of 400 mg administered by intravenous (IV) administration every 6 weeks trough within 20% of. In another embodiment, subcutaneous administration of an anti-PD-1 antibody or an antigen-binding fragment thereof results in the C of the antibody or the antigen-binding fragment trough at least the same as or greater than that of the anti-PD-1 antibody or an antigen-binding fragment thereof at a dose of 400 mg administered by intravenous (IV) administration every 6 weeks trough by less than 35%. In another embodiment, subcutaneous administration of an anti-PD-1 antibody or an antigen-binding fragment thereof results in the C of the antibody or the antigen-binding fragment trough at least the same as or greater than that of the anti-PD-1 antibody or an antigen-binding fragment thereof at a dose of 400 mg administered by intravenous (IV) administration every 6 weeks trough by less than 30%. In another embodiment, subcutaneous administration of an anti-PD-1 antibody or an antigen-binding fragment thereof results in the C of the antibody or the antigen-binding fragment trough being greater than that of the anti-PD-1 antibody or an antigen-binding fragment thereof at a dose of 400 mg administered by intravenous (IV) administration every 6 weeks trough by approximately 25-30%. In another embodiment, subcutaneous administration of an anti-PD-1 antibody or an antigen-binding fragment thereof results in the C of the antibody or the antigen-binding fragment trough being greater than that of the anti-PD-1 antibody or an antigen-binding fragment thereof at a dose of 400 mg administered by intravenous (IV) administration every 6 weeks trough by approximately 30%. In another embodiment, subcutaneous administration of an anti-PD-1 antibody or an antigen-binding fragment thereof results in the C of the antibody or the antigen-binding fragment trough being greater than that of the anti-PD-1 antibody or an antigen-binding fragment thereof at a dose of 400 mg administered by intravenous (IV) administration every 6 weeks trough by approximately 20-35%.

[0208] In certain embodiments of the methods or uses of the present invention, subcutaneous administration of a dose of an anti-PD-1 antibody (e.g., pembrolizumab) or an antigen-binding fragment thereof results in C trough equal to or higher than that of the dose administered via the 400 mg Q6W IV route. In embodiments of any of the methods or uses described herein, subcutaneous administration of an anti-PD-1 antibody or an antigen-binding fragment thereof results in subcutaneous C trough equal to or higher than that of the IV C trough with a ratio (e.g., geometric mean ratio) of at least 0.8, at least 1, at least 1.2, at least 1.3, at least 1.4, at least 1.5, or at least 1.6. In certain embodiments, subcutaneous administration results in PK properties with an SC:IV C trough ratio of at least 0.8 or higher. In certain embodiments, subcutaneous administration results in PK properties with an SC:IVC trough ratio of at least 1.0 or higher. In certain embodiments, subcutaneous administration results in an SC:IV C trough ratio of at least 1.2 or higher. In certain embodiments, subcutaneous administration results in an SC:IV C trough ratio of at least 1.3 or higher. In certain embodiments, subcutaneous administration results in an SC:IV C trough ratio of at least 1.4 or higher. In certain embodiments, subcutaneous administration results in an SC:IV C trough ratio of at least 1.5 or higher. In certain embodiments, subcutaneous administration results in an SC:IV C trough ratio of at least 1.6 or higher. trough ratio of at least 1.6 or higher.

[0209] In certain embodiments of the methods or uses of the present invention, subcutaneous administration of a dose of an anti-PD-1 antibody (e.g., pembrolizumab) or an antigen-binding fragment thereof results in an SC:IV C trough ratio of 0.8 to 1.6, 1.0 to 1.6, 1.1 to 1.6, 1.2 to 1.6, 1.3 to 1.6, 1.4 to 1.6, 1.2 to 1.5, 1.3 to 1.5, 1.4 to 1.5, or 1.3 to 1.4.

[0210] In certain embodiments of the methods or uses of the present invention, subcutaneous administration of a dose of an anti-PD-1 antibody (e.g., pembrolizumab) or an antigen-binding fragment thereof results in an SC:IV C trough ratio of 1.0 to 1.6. In certain embodiments, subcutaneous administration results in an SC:IV C trough ratio of 1.1 to 1.6. In certain embodiments, subcutaneous administration results in an SC:IVC troughThe ratio is 1.2 to 1.6. In certain embodiments, subcutaneous administration results in SC:IV C trough The ratio is 1.3 to 1.6. In certain embodiments, subcutaneous administration results in SC:IV C trough The ratio is 1.4 to 1.6. In certain embodiments, subcutaneous administration results in SC:IV C trough The ratio is 1.2 to 1.5. In certain embodiments, subcutaneous administration results in SC:IV C trough The ratio is 1.3 to 1.5. In certain embodiments, subcutaneous administration results in SC:IV C trough The ratio is 1.4 to 1.5. In certain embodiments, subcutaneous administration results in SC:IV C trough The ratio is 1.3 to 1.4. In certain embodiments, subcutaneous administration results in SC:IV C trough The ratio is 1.2 to 1.3. In certain embodiments, subcutaneous administration results in SC:IV C trough The ratio is 1.2 to 1.4.

[0211] In certain embodiments of the methods or uses of the present invention, subcutaneous administration of a dose of an anti-PD-1 antibody (e.g., pembrolizumab) or an antigen-binding fragment thereof results in an AUC that is at least 0.8 times the AUC of a 400 mg dose of an anti-PD-1 antibody or an antigen-binding fragment thereof administered by the Q6W IV route at cycle 1 or at steady state (0-6周) of the AUC (0-6周) . In certain embodiments of the methods or uses of the present invention, subcutaneous administration of a dose of an anti-PD-1 antibody (e.g., pembrolizumab) or an antigen-binding fragment thereof is an AUC that is approximately 1.0 times the AUC of a 400 mg dose of an anti-PD-1 antibody or an antigen-binding fragment thereof administered by the Q6W IV route during treatment (e.g., at cycle 1 or at steady state) (0-6周) of the AUC (0-6周) . In certain embodiments of the methods or uses of the present invention, subcutaneous administration of a dose of an anti-PD-1 antibody (e.g., pembrolizumab) or an antigen-binding fragment thereof results in an AUC that is approximately 0.8 to 1.2 times the AUC of a 400 mg dose of an anti-PD-1 antibody or an antigen-binding fragment thereof administered by the Q6W IV route during treatment (e.g., at cycle 1 or at steady state) (0-6周) of the AUC (0-6周) . In certain embodiments of the methods or uses of the present invention, subcutaneous administration of a dose of an anti-PD-1 antibody (e.g., pembrolizumab) or an antigen-binding fragment thereof results in an AUC that is approximately 1.0 to 1.1 times the AUC of a 400 mg dose of an anti-PD-1 antibody or an antigen-binding fragment thereof administered by the Q6W IV route during treatment (e.g., at cycle 1 or at steady state) (0-6周) of the AUC(0-6周) . In certain embodiments, after six cycles of administration, subcutaneous administration results in a PK profile with an SC:IV AUC of at least 0.8, 1.0 or higher (0-6周) ratio.

[0212] In an embodiment of any of the methods or uses of the present invention, subcutaneous administration of an anti-PD-1 antibody (e.g., pembrolizumab) co-formulated with human hyaluronidase (e.g., PH20 variant 2) at 760 - 790 mg every six weeks results in a C trough that is within 20% of the C trough of a 400 mg dose of the anti-PD-1 antibody administered by the intravenous (IV) route every 6 weeks. In a particular embodiment of the method or use of the present invention, subcutaneous administration of an anti-PD-1 antibody (e.g., pembrolizumab) co-formulated with human hyaluronidase (e.g., PH20 variant 2) at 760 - 790 mg every six weeks results in a subcutaneous C trough to IV C trough ratio (e.g., geometric mean ratio) of approximately 1.0 to 1.35 compared to the dose administered by the 400 mg Q6W IV route. In one embodiment of the foregoing embodiments, the subcutaneous administration of the anti-PD-1 antibody (e.g., pembrolizumab) is co-formulated with PH20 variant 2 and is 790 mg every six weeks. In a preferred embodiment of the method or use of the present invention, subcutaneous administration of an anti-PD-1 antibody (e.g., pembrolizumab) co-formulated with PH20 variant 2 at 790 mg every six weeks results in a subcutaneous C trough to IV C trough ratio (e.g., geometric mean ratio) of approximately 1.3 compared to the dose administered by the 400 mg Q6W IV route.

[0213] In a particular embodiment of the method or use of the present invention, subcutaneous administration of an anti-PD-1 antibody (e.g., pembrolizumab) co-formulated with human hyaluronidase (e.g., PH20 variant 2) at 760 - 790 mg every six weeks results in an AUC (0-6周) that is approximately 0.8 to 1.2 times the AUC (0-6周) of a 400 mg dose of the anti-PD-1 antibody administered by the Q6W IV route during the treatment duration. In a particular embodiment of the method or use of the present invention, subcutaneous administration of an anti-PD-1 antibody (e.g., pembrolizumab) co-formulated with human hyaluronidase (e.g., PH20 variant 2) at 760 - 790 mg every six weeks results in an AUC (0-6周) that is approximately 0.8 to 1.2 times the AUC (0-6周)a ratio of about 1.0 to 1.2. In certain embodiments of the methods or uses of the present invention, subcutaneous administration of an anti-PD-1 antibody (e.g., pembrolizumab) co-formulated with human hyaluronidase (e.g., PH20 variant 2) at 760 - 790 mg every six weeks results in an AUC (0-6周) that is a ratio of about 1.0 to 1.1 of the AUC (0-6周) for a 400 mg dose of the anti-PD-1 antibody administered via the Q6W IV route of administration over the duration of treatment. In one embodiment of the foregoing embodiments, subcutaneous administration of the anti-PD-1 antibody (e.g., pembrolizumab) co-formulated with PH20 variant 2 is 790 mg every six weeks. In a preferred embodiment of the methods or uses of the present invention, subcutaneous administration of an anti-PD-1 antibody (e.g., pembrolizumab) co-formulated with PH20 variant 2 at 790 mg every six weeks results in an AUC (0-6周) that is a ratio of about 1.1 of the AUC (0-6周) for the dose administered via the 400 mg Q6W IV route of administration.

[0214] In embodiments of any of the methods or uses of the present invention, subcutaneous administration of an anti-PD-1 antibody (e.g., pembrolizumab) co-formulated with human hyaluronidase (e.g., PH20 variant 2) at 760 - 790 mg every six weeks results in a C max that is approximately 40% lower than the C max for a 400 mg dose of the anti-PD-1 antibody administered via the intravenous (IV) route of administration every six weeks in cycle 1. In embodiments of any of the methods or uses of the present invention, subcutaneous administration of an anti-PD-1 antibody (e.g., pembrolizumab) co-formulated with human hyaluronidase (e.g., PH20 variant 2) at 760 - 790 mg every six weeks results in a C max that is approximately 35 - 38% lower than the C max for a 400 mg dose of the anti-PD-1 antibody administered via the intravenous (IV) route of administration every six weeks in cycle 1. In embodiments of any of the methods or uses of the present invention, subcutaneous administration of an anti-PD-1 antibody (e.g., pembrolizumab) co-formulated with human hyaluronidase (e.g., PH20 variant 2) at 760 - 790 mg every six weeks results in a C max that is approximately 20 - 30% lower than the C max for a 400 mg dose of the anti-PD-1 antibody or antigen-binding fragment thereof administered via the intravenous (IV) route of administration every six weeks at steady state. In embodiments of any of the methods or uses of the present invention, subcutaneous administration of an anti-PD-1 antibody (e.g., pembrolizumab) co-formulated with human hyaluronidase (e.g., PH20 variant 2) at 760 - 790 mg every six weeks results in a C max, which is about 22 - 25% lower than the C of a 400 mg dose of anti-PD-1 antibody administered via the intravenous (IV) route every 6 weeks at steady state. In one embodiment of the foregoing embodiments, the subcutaneous administration of an anti-PD-1 antibody (e.g., pembrolizumab) co-formulated with human hyaluronidase (e.g., PH20 variant 2) is 790 mg every six weeks. In an embodiment of any of the methods or uses of the present invention, subcutaneous administration of an anti-PD-1 antibody (e.g., pembrolizumab) co-formulated with PH20 variant 2 at 790 mg every six weeks results in a C max , which is about 22% lower than the C of a 400 mg dose of anti-PD-1 antibody administered via the intravenous (IV) route every 6 weeks at steady state. In an embodiment of any of the methods or uses of the present invention, subcutaneous administration of an anti-PD-1 antibody (e.g., pembrolizumab) co-formulated with PH20 variant 2 at 790 mg every six weeks results in a C max , which is about 22% lower than the C of a 400 mg dose of anti-PD-1 antibody administered via the intravenous (IV) route every 6 weeks at steady state. In an embodiment of any of the methods or uses of the present invention, subcutaneous administration of an anti-PD-1 antibody (e.g., pembrolizumab) co-formulated with PH20 variant 2 at 790 mg every six weeks results in a C max , which is about 35% lower than the C of a 400 mg dose of anti-PD-1 antibody administered via the intravenous (IV) route every 6 weeks at cycle 1. max , which is about 35% lower than the C of a 400 mg dose of anti-PD-1 antibody administered via the intravenous (IV) route every 6 weeks at cycle 1. max In an embodiment of any of the methods or uses of the present invention, subcutaneous administration of an anti-PD-1 antibody (e.g., pembrolizumab) co-formulated with human hyaluronidase (e.g., PH20 variant 2) at 380 - 395 mg every three weeks results in a C

[0215] , which is within 20% of the C of a 400 mg dose of anti-PD-1 antibody administered via the intravenous (IV) route every 6 weeks. In a particular embodiment of the methods or uses of the present invention, subcutaneous administration of an anti-PD-1 antibody (e.g., pembrolizumab) co-formulated with human hyaluronidase (e.g., PH20 variant 2) at 380 - 395 mg every three weeks results in a subcutaneous C trough to IV C trough ratio (e.g., geometric mean ratio) of about 1.0 to 2.0 of the dose administered via the 400 mg Q6W IV route. In one embodiment of the foregoing embodiments, the subcutaneous administration of an anti-PD-1 antibody (e.g., pembrolizumab) is co-formulated with PH20 variant 2 and is 395 mg every three weeks. In a preferred embodiment of the methods or uses of the present invention, subcutaneous administration of an anti-PD-1 antibody (e.g., pembrolizumab) co-formulated with PH20 variant 2 at 395 mg every three weeks results in a subcutaneous C trough to IV C trough ratio (e.g., geometric mean ratio) of about 1.5 - 2.0 of the dose administered via the 400 mg Q6W IV route. trough to IV C trough ratio (e.g., geometric mean ratio).

[0216] In certain embodiments of the methods or uses of the present invention, subcutaneous administration of an anti-PD-1 antibody (e.g., pembrolizumab) co-formulated with human hyaluronidase (e.g., PH20 variant 2) at 380 - 395 mg every three weeks results in an AUC (0-6周) that is about 0.8 to 1.2 times the AUC (0-6周) of a 400 mg dose of the anti-PD-1 antibody administered via the Q6W IV route during the treatment duration. In certain embodiments of the methods or uses of the present invention, subcutaneous administration of an anti-PD-1 antibody (e.g., pembrolizumab) co-formulated with human hyaluronidase (e.g., PH20 variant 2) at 380 - 395 mg every three weeks results in an AUC (0-6周) that is about 0.8 to 1.1 times the AUC (0-6周) of a 400 mg dose of the anti-PD-1 antibody administered via the Q6W IV route during the treatment duration. In one embodiment of the foregoing embodiments, the subcutaneous administration of the anti-PD-1 antibody (e.g., pembrolizumab) co-formulated with PH20 variant 2 is 395 mg every three weeks. In a preferred embodiment of the methods or uses of the present invention, subcutaneous administration of an anti-PD-1 antibody (e.g., pembrolizumab) co-formulated with PH20 variant 2 at 395 mg every three weeks results in an AUC (0-6周) that is about 1.1 times the AUC (0-6周) of the dose administered via the 400 mg Q6W IV route.

[0217] In one embodiment of the foregoing embodiments, the ratio is the geometric mean ratio. In one embodiment of the foregoing embodiments, the ratio is at Cycle 1 (3 weeks or 6 weeks, depending on the subcutaneous administration cycle). In one embodiment of the foregoing embodiments, the ratio is at steady state. In one embodiment of the foregoing embodiments, the ratio is during the treatment duration.

[0218] In certain embodiments of the methods or uses of the present invention, the cancer is selected from the following: melanoma, lung cancer, head and neck cancer, bladder cancer, breast cancer, gastrointestinal cancer, multiple myeloma, hepatocellular carcinoma, Merkel cell carcinoma, cutaneous squamous cell carcinoma, lymphoma, renal cancer, mesothelioma, ovarian cancer, esophageal cancer, anal cancer, cholangiocarcinoma, colorectal cancer, endometrial cancer, cervical cancer, thyroid cancer, salivary gland cancer, prostate cancer (e.g., hormone-refractory prostate cancer), pancreatic cancer, colon cancer, liver cancer, thyroid cancer, glioblastoma, glioma, and other neoplastic malignancies.

[0219] In certain embodiments, the lung cancer is non-small cell lung cancer.

[0220] In certain embodiments, the lung cancer is small cell lung cancer.

[0221] In certain embodiments, the lymphoma is Hodgkin lymphoma.

[0222] In certain embodiments, the lymphoma is non-Hodgkin lymphoma. In certain embodiments, the lymphoma is primary mediastinal large B-cell lymphoma (PMBCL). In certain embodiments, the lymphoma is diffuse large B-cell lymphoma (DLBCL). In certain embodiments, the lymphoma is mantle cell lymphoma.

[0223] In certain embodiments, the breast cancer is triple-negative breast cancer.

[0224] In certain embodiments, the breast cancer is ER+ / HER2- breast cancer.

[0225] In certain embodiments, the breast cancer is HR+ / HER2- breast cancer.

[0226] In certain embodiments, the breast cancer is HER2+ breast cancer.

[0227] In certain embodiments, the breast cancer is ER+ breast cancer.

[0228] In certain embodiments, the breast cancer is germline BRCA-mutated HER2- breast cancer.

[0229] In certain embodiments, the bladder cancer is urothelial carcinoma.

[0230] In certain embodiments, the head and neck cancer is nasopharyngeal carcinoma. In certain embodiments, the cancer is thyroid cancer. In other embodiments, the cancer is salivary gland cancer. In other embodiments, the cancer is head and neck squamous cell carcinoma.

[0231] In certain embodiments, the cancer is metastatic colorectal cancer with high microsatellite instability (MSI-H).

[0232] In certain embodiments, the cancer is microsatellite stable (MSS) colorectal cancer.

[0233] In certain embodiments, the cancer is solid tumor with high microsatellite instability (MSI-H).

[0234] In certain embodiments, the cancer is solid tumor with high mutational burden.

[0235] In a particular embodiment of the method or use of the present invention, the cancer is selected from the following: melanoma, non-small cell lung cancer, small cell lung cancer, head and neck squamous cell carcinoma, classical Hodgkin lymphoma, primary mediastinal large B-cell lymphoma, urothelial carcinoma, microsatellite instability-high or mismatch repair-deficient cancer, gastric cancer, esophageal cancer, cervical cancer, hepatocellular carcinoma, Merkel cell carcinoma, renal cell carcinoma, endometrial cancer, cancer characterized by a tumor with a high mutation burden, cutaneous squamous cell carcinoma, and triple-negative breast cancer.

[0236] In a particular embodiment of the method or use of the present invention, the cancer is selected from the following: melanoma, non-small cell lung cancer, head and neck squamous cell carcinoma, classical Hodgkin lymphoma, primary mediastinal large B-cell lymphoma, urothelial carcinoma, microsatellite instability-high or mismatch repair-deficient cancer, gastric cancer, esophageal cancer, cervical cancer, hepatocellular carcinoma, Merkel cell carcinoma, renal cell carcinoma, endometrial cancer, cancer characterized by a tumor with a high mutation burden, cutaneous squamous cell carcinoma, and triple-negative breast cancer.

[0237] In a particular embodiment of the method or use of the present invention, the cancer is selected from the following: melanoma, non-small cell lung cancer, head and neck squamous cell carcinoma, urothelial carcinoma, classical Hodgkin lymphoma, primary mediastinal large B-cell lymphoma (PMBCL), MSI-H cancer, MSI-H or mismatch repair-deficient colorectal cancer, gastric cancer, gastroesophageal junction adenocarcinoma, esophageal cancer, cervical cancer, hepatocellular carcinoma, Merkel cell carcinoma, renal cell carcinoma, endometrial cancer, cutaneous squamous cell carcinoma, cancer with a high tumor mutational burden (TMB-H), and triple-negative breast cancer.

[0238] In a first embodiment (Embodiment E1), the present invention comprises a method of treating cancer in a human patient in need thereof, which comprises subcutaneously administering to the patient a dose of about 600 mg to about 1000 mg of an anti-PD-1 antibody (e.g., pembrolizumab) or an antigen-binding fragment thereof approximately once every six weeks. In a particular embodiment, the anti-PD-1 antibody or an antigen-binding fragment thereof is administered once every six weeks.

[0239] In a second embodiment (Embodiment E2), the present invention includes a method of treating unresectable or metastatic melanoma in a human patient in need thereof, comprising subcutaneously administering to the patient a dose of about 600 mg to about 1000 mg of an anti-PD-1 antibody (e.g., pembrolizumab) or an antigen-binding fragment thereof about once every six weeks. In certain embodiments, the anti-PD-1 antibody or an antigen-binding fragment thereof is administered once every six weeks. In a third embodiment (Embodiment E3), the present invention includes a method of treating metastatic non-small cell lung cancer (NSCLC) in a human patient in need thereof, comprising subcutaneously administering to the patient a dose of about 600 mg to about 1000 mg of an anti-PD-1 antibody (e.g., pembrolizumab) or an antigen-binding fragment thereof about once every six weeks. In certain embodiments, the anti-PD-1 antibody or an antigen-binding fragment thereof is administered once every six weeks.

[0240] In a sub-embodiment of Embodiment E3 (Embodiment E3-A), the patient has a tumor with high PD-L1 expression [(tumor proportion score (TPS) ≥ 50%)] and has not previously been treated with platinum-containing chemotherapy.

[0241] In a further sub-embodiment of Embodiment E3 (Embodiment E3-B), the patient has a tumor with PD-L1 expression (TPS ≥ 1%) and has previously been treated with platinum-containing chemotherapy. In certain embodiments of Embodiment E3-B, the patient had disease progression during or after receiving platinum-containing chemotherapy or at least one prior chemotherapy.

[0242] In another sub-embodiment of Embodiment E3 (Embodiment E3-C), the patient has a tumor with PD-L1 expression (TPS ≥ 1%) and has not previously been treated with platinum-containing chemotherapy. In one embodiment, the patient has stage III non-small cell lung cancer and is not suitable for surgical resection or definitive chemoradiation. In one embodiment, the patient has metastatic non-small cell lung cancer. In yet another sub-embodiment of Embodiment E3 (Embodiment E3-D), the PD-L1 expression of the patient's tumor is not detected. In this embodiment, the patient is treated with an anti-PD-1 antibody or an antigen-binding fragment thereof regardless of PD-L1 expression. In certain embodiments, the patient has not previously been treated with platinum-containing chemotherapy.

[0243] In certain embodiments of Embodiment E3 (including Embodiment E3-A, E3-B, and E3-C), the PD-L1 TPS is determined by an FDA-approved assay.

[0244] In certain embodiments of Embodiment E3 (including Embodiment E3-A, E3-B, E3-C, and E3-D), the patient's tumor does not have EGFR or ALK genomic aberrations.

[0245] In certain embodiments of Embodiment E3 (including Embodiments E3-A, E3-B, E3-C, and E3-D), the patient's tumor has an EGFR or ALK genomic abnormality and has disease progression during or after receiving treatment for the EGFR or ALK abnormality prior to receiving an anti-PD-1 antibody or an antigen-binding fragment thereof.

[0246] In a fourth embodiment (Embodiment E4), the present invention includes a method of treating metastatic or stage III non-small cell lung cancer (NSCLC) in a human patient in need thereof, comprising: (1) subcutaneously administering to the patient a dose of from about 600 mg to about 1000 mg of an anti-PD-1 antibody (e.g., pembrolizumab) or an antigen-binding fragment thereof approximately once every six weeks, and (2) administering to the patient pemetrexed and platinum chemotherapy (e.g., carboplatin). In a sub-embodiment of Embodiment E4, the patient has not previously been treated with an anti-cancer therapeutic agent prior to initiating the combination treatment regimen of the anti-PD-1 antibody or an antigen-binding fragment thereof, pemetrexed, and carboplatin.

[0247] In certain embodiments of Embodiments E3 and E4 (including sub-embodiments thereof), the patient has non-small cell lung cancer.

[0248] In certain embodiments of Embodiments E3 and E4 (including sub-embodiments thereof), the patient is also treated with carboplatin and paclitaxel or albumin-bound paclitaxel. In one embodiment, carboplatin is administered by intravenous infusion at an AUC of 5-6 mg / ml / min, paclitaxel is administered by intravenous infusion at 200 mg / m 2 every 21 days, and albumin-bound paclitaxel is administered by intravenous infusion at 100 mg / m 2 every 7 days.

[0249] In a sub-embodiment of Embodiment E4, pemetrexed is administered to the patient in an amount of 500 mg / m 2 .

[0250] In a sub-embodiment of Embodiment E4, pemetrexed is administered to the patient in an amount of 500 mg / m 2 every 3 weeks.

[0251] In a sub-embodiment of Embodiment E4, pemetrexed is administered to the patient by intravenous infusion every 21 days. In a particular embodiment, the infusion time is about 10 minutes.

[0252] In a sub - embodiment of Embodiment E4 (Embodiment E4 - A), the present invention further includes administering to a patient about 400 μg to about 1000 μg of folic acid once a day, starting about 7 days before administering pemetrexed to the patient and continuing until about 21 days after administering the last dose of pemetrexed to the patient. In certain embodiments, the folic acid is administered orally.

[0253] In a sub - embodiment of Embodiments E4 and E4 - A (Embodiment E4 - B), the present invention further includes administering to a patient about 1 mg of vitamin B about 1 week before the first administration of pemetrexed and about every three cycles of pemetrexed administration (i.e., about every 9 weeks). 12 In certain embodiments, the vitamin B is administered intramuscularly. 12 .

[0254] In a sub - embodiment of Embodiments E4, E4 - A, and E4 - B (Embodiment E4 - C), the present invention further includes administering to a patient about 4 mg of dexamethasone twice a day on the day before, the day of, and the day after pemetrexed administration. In certain embodiments, the dexamethasone is administered orally. In a fifth embodiment (Embodiment E5), the present invention includes a method of treating recurrent or metastatic head and neck squamous cell carcinoma (HNSCC) in a human patient in need thereof, which includes subcutaneously administering to the patient a dose of about 600 mg to about 1000 mg of an anti - PD - 1 antibody (e.g., pembrolizumab) or an antigen - binding fragment thereof about once every six weeks.

[0255] In a sub - embodiment of Embodiment E5 (Embodiment E5 - A), the patient was previously treated with platinum - containing chemotherapy. In certain embodiments, the patient has disease progression during or after platinum - containing chemotherapy.

[0256] In a sub - embodiment of Embodiment E5 (Embodiment E5 - B), the patient has metastatic or unresectable recurrent HNSCC, and the method further includes administering platinum and 5 - FU (fluorouracil) for first - line treatment of HNSCC.

[0257] In a sub - embodiment of Embodiment E5 (Embodiment E5 - C), the anti - PD - 1 antibody (e.g., pembrolizumab) is administered as a single agent for first - line treatment of patients with metastatic or unresectable recurrent HNSCC, wherein the patient's tumor expresses PD - L1 (CPS≥1%).

[0258] In a sub - embodiment of Embodiment E5 (Embodiment E5 - D), the anti - PD - 1 antibody (e.g., pembrolizumab) is administered in combination with platinum and 5 - FU (fluorouracil) chemotherapy to a patient for first - line treatment of metastatic or unresectable recurrent head and neck squamous cell carcinoma, wherein the patient's tumor expresses PD - L1 and has a CPS≥1.

[0259] In a sub - embodiment of Embodiment E5 (Embodiment E5 - E), an anti - PD - 1 antibody (e.g., pembrolizumab) is administered for the treatment of metastatic or unresectable recurrent head and neck squamous cell carcinoma in a patient, wherein the patient's tumor expresses PD - L1, has a ≥50% TPS, and has progressed during or after platinum - containing chemotherapy.

[0260] In the embodiments of Embodiment E5 - B and E5 - D, the platinum - containing chemotherapy is carboplatin administered by intravenous infusion at an AUC of 5 mg / ml / min every three weeks, or cisplatin administered by intravenous infusion at 100 mg / m 2 administered, and 5 - FU administered at 1000 mg / m 2 / day for 4 consecutive days every three weeks.

[0261] In a sixth embodiment (Embodiment E6), the present invention includes a method for treating refractory or recurrent classical Hodgkin lymphoma (cHL) in a human patient in need thereof, which comprises subcutaneously administering to the patient a dose of about 600 mg to about 1000 mg of an anti - PD - 1 antibody (e.g., pembrolizumab) or an antigen - binding fragment thereof approximately once every six weeks.

[0262] In a seventh embodiment (Embodiment E7), the present invention includes a method for treating classical Hodgkin lymphoma (cHL) in a human patient in need thereof, which comprises subcutaneously administering to the patient a dose of about 600 mg to about 1000 mg of an anti - PD - 1 antibody (e.g., pembrolizumab) or an antigen - binding fragment thereof approximately once every six weeks, wherein the patient relapses after (a) first - line or multiple - line therapy for cHL, (b) second - line or higher - line therapy for cHL, or (c) third - line or higher - line therapy for cHL.

[0263] In sub - embodiments of Embodiment E6 and E7, the patient is an adult human patient.

[0264] In alternative sub - embodiments of Embodiment E6 and E7, the patient is a pediatric patient.

[0265] In an eighth embodiment (Embodiment E8), the present invention includes a method for treating locally advanced or metastatic urothelial carcinoma in a human patient in need thereof, which comprises subcutaneously administering to the patient a dose of about 600 mg to about 1000 mg of an anti - PD - 1 antibody (e.g., pembrolizumab) or an antigen - binding fragment thereof approximately once every six weeks. In one embodiment, the patient is ineligible for platinum - containing chemotherapy or has disease progression during or after platinum - containing chemotherapy or within 12 months of neoadjuvant or adjuvant treatment with platinum - containing chemotherapy.

[0266] In a sub - embodiment of Embodiment E8, the patient is ineligible for cisplatin - containing chemotherapy.

[0267] In a sub - embodiment of Embodiment E8, the patient has disease progression during or after platinum - containing chemotherapy or within 12 months of neoadjuvant or adjuvant treatment with platinum - containing chemotherapy.

[0268] In a sub - embodiment of Embodiment E8, the patient's tumor expresses PD - L1. In other sub - embodiments of Embodiment E8, the patient's tumor expresses PD - L1 (CPS≥10).

[0269] In a ninth embodiment (Embodiment E9), the present invention includes a method for treating unresectable or metastatic microsatellite instability - high (MSI - H) or mismatch repair (MMR) - defective solid tumors in a human patient in need thereof, comprising subcutaneously administering to the patient a dose of about 600 mg to about 1000 mg of an anti - PD - 1 antibody (e.g., pembrolizumab) or an antigen - binding fragment thereof approximately once every six weeks.

[0270] In a sub - embodiment of Embodiment E9, the patient has disease progression after prior anti - cancer treatment.

[0271] In a sub - embodiment of Embodiment E9, the patient has advanced or recurrent endometrial cancer. In one embodiment, there is disease progression during or after prior treatment with platinum - containing therapy and the patient is not suitable for curative surgery or radiotherapy.

[0272] In a sub - embodiment of Embodiment E9, the patient has unresectable or metastatic gastric cancer, small intestine cancer, or cholangiocarcinoma. In one embodiment, the patient has disease progression during or after at least one prior therapy.

[0273] In a tenth embodiment (Embodiment E10), the present invention includes a method for treating unresectable or metastatic MSI - H or MMR - defective colorectal cancer in a human patient in need thereof, comprising subcutaneously administering to the patient a dose of about 600 mg to about 1000 mg of an anti - PD - 1 antibody (e.g., pembrolizumab) or an antigen - binding fragment thereof approximately once every six weeks.

[0274] In a sub - embodiment of Embodiment E10, the patient has disease progression after prior use of fluoropyrimidine, oxaliplatin, and irinotecan.

[0275] In the eleventh embodiment (Embodiment E11), the present invention includes a method for treating recurrent locally advanced unresectable or metastatic gastric cancer or gastroesophageal junction adenocarcinoma in a human patient in need thereof, which comprises subcutaneously administering to the patient a dose of about 600 mg to about 1000 mg of an anti-PD-1 antibody (e.g., pembrolizumab) or an antigen-binding fragment thereof about once every six weeks. In a particular embodiment, the gastric cancer or gastroesophageal junction adenocarcinoma is HER2-positive. In a particular embodiment, the present invention further includes treating the patient with trastuzumab, fluoropyrimidine, and platinum-containing chemotherapy. In a particular embodiment, the treatment with the anti-PD-1 antibody, trastuzumab, fluoropyrimidine, and platinum-containing chemotherapy is first-line treatment. In one embodiment, trastuzumab is administered at 8 mg / kg at the first infusion and at 6 mg / kg in subsequent cycles, followed by chemotherapy: cisplatin 80 mg / m 2 for up to 6 cycles and 5-FU 800 mg / m 2 / day for 5 days or oxaliplatin 130 mg / m 2 for up to 6 - 8 cycles, each administered every three weeks and capecitabine 1,000 mg / m 2 twice daily for 14 days. In one embodiment, the anti-PD-1 antibody (e.g., pembrolizumab) administered subcutaneously every 6 weeks is administered on day 1 of each cycle before trastuzumab and chemotherapy.

[0276] In the twelfth embodiment (Embodiment E12), the present invention includes a method for treating recurrent locally advanced or metastatic esophageal cancer or gastroesophageal junction adenocarcinoma in a human patient in need thereof, which comprises subcutaneously administering to the patient a dose of about 600 mg to about 1000 mg of an anti-PD-1 antibody (e.g., pembrolizumab) or an antigen-binding fragment thereof about once every six weeks.

[0277] In a sub-embodiment of Embodiment E12, the method further comprises combinatorial administration of fluoropyrimidine-based chemotherapy and platinum-containing chemotherapy.

[0278] In another twelfth embodiment (Embodiment E12), the present invention includes a method for treating recurrent locally advanced or metastatic esophageal cancer or HER-2 negative gastroesophageal junction adenocarcinoma in a human patient in need thereof, which comprises subcutaneously administering to the patient a dose of about 600 mg to about 1000 mg of an anti-PD-1 antibody (e.g., pembrolizumab) or an antigen-binding fragment thereof in combination with fluoropyrimidine-based chemotherapy and platinum-containing chemotherapy, and the patient's tumor has a PD-L1 combined positive score (CPS) ≥ 10.

[0279] In a sub-embodiment of Embodiment E12, the fluoropyrimidine-based chemotherapy and platinum-containing chemotherapy are cisplatin 80 mg / m 2For up to 6 cycles and 5-FU 800 mg / m 2 / day for 5 days or oxaliplatin 130 mg / m 2 For up to 6 - 8 cycles, each administered every three weeks and capecitabine 1,000 mg / m 2 Twice daily for 14 days.

[0280] In sub - embodiments of embodiments E11 and E12, the patient's tumor expresses PD - L1. In sub - embodiments of embodiments E11 and E12, the patient's tumor has a PD - L1 combined positive score (CPS) ≥1.

[0281] In sub - embodiments of embodiments E11 and E12, the patient has disease progression at or after one or more prior lines of therapy. In certain embodiments, the prior lines of therapy include fluoropyrimidine and platinum - containing chemotherapy.

[0282] In sub - embodiments of embodiments E11 and E12, the patient has disease progression at or after two or more prior lines of therapy, said lines of therapy including fluoropyrimidine - containing chemotherapy and platinum - containing chemotherapy.

[0283] In sub - embodiments of embodiments E11 and E12, the patient has disease progression at or after one or more prior lines of therapy, said lines of therapy including HER2 / neu - targeted therapy.

[0284] In sub - embodiments of embodiments E11 and E12, the patient has disease progression at or after two or more prior lines of therapy, said lines of therapy including HER2 / neu - targeted therapy.

[0285] In a thirteenth embodiment (embodiment E13), the present invention includes a method of treating cancer in a human patient in need thereof, comprising subcutaneously administering to the patient a dose of about 600 mg to about 1,000 mg of an anti - PD - 1 antibody (e.g., pembrolizumab) or an antigen - binding fragment thereof approximately once every six weeks, wherein the patient has a cancer selected from the group consisting of melanoma, lung cancer, head and neck cancer, bladder cancer, breast cancer, gastrointestinal cancer, multiple myeloma, hepatocellular carcinoma, lymphoma, kidney cancer, mesothelioma, ovarian cancer, esophageal cancer, anal cancer, cholangiocarcinoma, colorectal cancer, cervical cancer, hepatocellular carcinoma, Merkel cell carcinoma, renal cell carcinoma, endometrial cancer, cutaneous squamous cell carcinoma, thyroid cancer, and salivary gland cancer.

[0286] In a fourteenth embodiment (Embodiment E14), the invention includes a method of treating cancer in a human patient in need thereof, comprising subcutaneously administering to the patient a dose of from about 600 mg to about 1000 mg of an anti-PD-1 antibody (e.g., pembrolizumab) or an antigen-binding fragment thereof, about once every six weeks, wherein the patient has small cell lung cancer. In a sub-embodiment, the patient was previously treated with platinum-based chemotherapy and at least one other prior line of therapy.

[0287] In a fifteenth embodiment (Embodiment E15), the invention includes a method of treating non-Hodgkin lymphoma in a human patient in need thereof, comprising subcutaneously administering to the patient a dose of from about 600 mg to about 1000 mg of an anti-PD-1 antibody (e.g., pembrolizumab) or an antigen-binding fragment thereof, about once every six weeks.

[0288] In a sub-embodiment of Embodiment 15, the non-Hodgkin lymphoma is primary mediastinal large B-cell lymphoma (PMBCL). In a particular embodiment, wherein the patient has PMBCL, the patient has refractory PMBCL. In a particular embodiment, the patient has relapsed after one or more prior lines of therapy. In a particular embodiment, the patient has relapsed after two or more prior lines of therapy. In a particular embodiment, the patient has not been previously treated with another line of therapy. In a particular embodiment, the patient is an adult. In a particular embodiment, the patient is a pediatric patient.

[0289] In a sixteenth embodiment (Embodiment E16), the invention includes a method of treating metastatic squamous NSCLC in a human patient in need thereof, comprising: (1) subcutaneously administering to the patient a dose of from about 600 mg to about 1000 mg of an anti-PD-1 antibody (e.g., pembrolizumab) or an antigen-binding fragment thereof, about once every six weeks, and (2) administering to the patient (i) carboplatin and paclitaxel, or (ii) carboplatin and albumin-bound paclitaxel.

[0290] In a seventeenth embodiment (Embodiment E17), the invention includes a method of treating Merkel cell carcinoma (MCC) in a human patient in need thereof, comprising subcutaneously administering to the patient a dose of from about 600 mg to about 1000 mg of an anti-PD-1 antibody (e.g., pembrolizumab) or an antigen-binding fragment thereof, about once every six weeks. In a particular sub-embodiment of Embodiment E17, the cancer is recurrent locally advanced MCC. In a particular sub-embodiment of Embodiment E17, the cancer is metastatic MCC.

[0291] In a sub-embodiment of Embodiment E17, the patient is an adult human patient. In an alternative sub-embodiment of Embodiment E17, the patient is a pediatric patient.

[0292] In the eighteenth embodiment (Embodiment E18), the present invention includes a method for adjuvant treatment of melanoma in a human patient in need thereof, which comprises subcutaneously administering to the patient a dose of about 600 mg to about 1000 mg of an anti-PD-1 antibody (e.g., pembrolizumab) or an antigen-binding fragment thereof approximately once every six weeks, wherein the patient has previously had one or more melanoma lesions resected. In a sub-embodiment of Embodiment E18, the method comprises treating resected high-risk stage III melanoma. In a sub-embodiment of Embodiment E18, the method comprises treating resected stage IIB or IIC melanoma.

[0293] In the nineteenth embodiment (Embodiment E19), the present invention includes a method for treating hepatocellular carcinoma (HCC) in a human patient in need thereof, which comprises subcutaneously administering to the patient a dose of about 600 mg to about 1000 mg of an anti-PD-1 antibody (e.g., pembrolizumab) or an antigen-binding fragment thereof approximately once every six weeks. In a particular embodiment of Embodiment E19, the patient has previously been treated with sorafenib.

[0294] In the twentieth embodiment (Embodiment E20), the present invention includes a method for treating renal cell carcinoma (RCC) in a human patient in need thereof, which comprises subcutaneously administering to the patient a dose of about 600 mg to about 1000 mg of an anti-PD-1 antibody (e.g., pembrolizumab) or an antigen-binding fragment thereof approximately once every six weeks.

[0295] In a sub-embodiment of Embodiment E20, the cancer is advanced clear cell RCC.

[0296] In a sub-embodiment of Embodiment E20, the patient has advanced or metastatic renal cell carcinoma (RCC).

[0297] In a sub-embodiment of Embodiment E20 (Embodiment E20C), the anti-PD-1 antibody (e.g., pembrolizumab) is for adjuvant treatment of RCC patients with medium-high or high recurrence risk after nephrectomy or after nephrectomy and resection of metastatic lesions.

[0298] In the twenty-first embodiment (Embodiment E21), the present invention includes a method for treating breast cancer in a human patient in need thereof, which comprises subcutaneously administering to the patient a dose of about 600 mg to about 1000 mg of an anti-PD-1 antibody (e.g., pembrolizumab) or an antigen-binding fragment thereof approximately once every six weeks.

[0299] In a sub - embodiment of embodiment E21, the breast cancer is triple - negative breast cancer. In a further sub - embodiment, the patient is further treated with chemotherapy. In a further sub - embodiment, the TNBC is recurrent unresectable or metastatic TNBC and the patient's tumor expresses PD - L1 (CPS≥10).

[0300] In a sub - embodiment of embodiment E21, the breast cancer is ER + / HER2 - breast cancer.

[0301] In a further sub - embodiment of embodiment E21, the patient has high - risk early or locally advanced TNBC, and the method comprises treating the patient with a combination of an anti - PD - 1 antibody (e.g., pembrolizumab) and chemotherapy as neoadjuvant therapy, and then treating the patient with an anti - PD - 1 antibody (e.g., pembrolizumab) as a single agent as adjuvant therapy after surgery. In one embodiment, the neoadjuvant anti - PD - 1 antibody (e.g., pembrolizumab) is administered to the patient on day 1 of cycles 1 - 4 of the treatment regimen for four cycles in combination with: carboplatin administered every 3 weeks at AUC 5mg / mL / min on day 1 of cycles 1 - 4 of the treatment regimen or at AUC 1.5mg / mL / min on days 1, 8, and 15 of cycles 1 - 4 of the treatment regimen, and paclitaxel 80mg / m 2 ; followed by an additional four cycles of neoadjuvant anti - PD - 1 antibody (e.g., pembrolizumab) on day 1 of cycles 5 - 8 of the treatment regimen in combination with: doxorubicin 60mg / m 2 or epirubicin 90mg / m 2 and cyclophosphamide 600mg / m every 3 weeks on day 1 of cycles 5 - 8 of the treatment regimen 2 ; and after surgery, nine cycles of adjuvant anti - PD - 1 antibody (e.g., pembrolizumab).

[0302] In the twenty - second embodiment (embodiment E22), the present invention includes a method of treating nasopharyngeal cancer in a human patient in need thereof, which comprises subcutaneously administering to the patient a dose of about 600mg to about 1000mg of an anti - PD - 1 antibody (e.g., pembrolizumab) or an antigen - binding fragment thereof approximately once every six weeks.

[0303] In the twenty - third embodiment (embodiment E23), the present invention includes a method of treating thyroid cancer in a human patient in need thereof, which comprises subcutaneously administering to the patient a dose of about 600mg to about 1000mg of an anti - PD - 1 antibody (e.g., pembrolizumab) or an antigen - binding fragment thereof approximately once every six weeks.

[0304] In the twenty-fourth embodiment (Embodiment E24), the present invention includes a method for treating salivary gland cancer in a human patient in need thereof, which comprises subcutaneously administering to the patient a dose of about 600 mg to about 1000 mg of an anti-PD-1 antibody (e.g., pembrolizumab) or an antigen-binding fragment thereof about once every six weeks.

[0305] In the twenty-fifth embodiment (Embodiment E25), the present invention includes a method for treating cancer in a human patient in need thereof, which comprises subcutaneously administering to the patient a dose of about 600 mg to about 1000 mg of an anti-PD-1 antibody (e.g., pembrolizumab) or an antigen-binding fragment thereof about once every six weeks, wherein the cancer is selected from the following: melanoma, non-small cell lung cancer (NSCLC), head and neck squamous cell carcinoma (HNSCC), recurrent or refractory classical Hodgkin lymphoma (cHL), primary mediastinal large B-cell lymphoma (PMBCL), urothelial carcinoma, microsatellite instability-high or mismatch repair-deficient colorectal cancer, gastric cancer, esophageal cancer, cervical cancer, hepatocellular carcinoma, Merkel cell carcinoma, renal cell carcinoma, endometrial cancer, TMB-H cancer, cutaneous squamous cell carcinoma, and triple-negative breast cancer.

[0306] In a sub-embodiment of Embodiment E25 (Embodiment 25B), the present invention includes a method for treating cancer in a human patient in need thereof, which comprises subcutaneously administering to the patient a dose of about 600 mg to about 1000 mg of an anti-PD-1 antibody (e.g., pembrolizumab) or an antigen-binding fragment thereof about once every six weeks, wherein the cancer is selected from the following: melanoma, non-small cell lung cancer, recurrent or refractory classical Hodgkin lymphoma, primary mediastinal large B-cell lymphoma, head and neck squamous cell carcinoma, urothelial carcinoma, esophageal cancer, gastric cancer, cervical cancer, PMBCL, MSI-H cancer, hepatocellular carcinoma, Merkel cell carcinoma, renal cell carcinoma, endometrial cancer, TMB-H cancer, cutaneous squamous cell carcinoma, and triple-negative breast cancer.

[0307] In the twenty-sixth embodiment (Embodiment E26), the present invention includes a method for treating cancer in a human patient in need thereof, which comprises subcutaneously administering to the patient a dose of about 600 mg to about 1000 mg of an anti-PD-1 antibody (e.g., pembrolizumab) or an antigen-binding fragment thereof about once every six weeks, wherein the cancer is a stromal core tumor.

[0308] In a sub - embodiment of Embodiment E26, the hematopoietic malignancies are selected from the following: acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML), diffuse large B - cell lymphoma (DLBCL), EBV - positive DLBCL, primary mediastinal large B - cell lymphoma, T - cell / histiocyte - rich large B - cell lymphoma, follicular lymphoma, Hodgkin's lymphoma (HL), mantle cell lymphoma (MCL), multiple myeloma (MM), myeloid leukemia - 1 protein (MCL - 1), myelodysplastic syndrome (MDS), non - Hodgkin lymphoma (NHL), and small lymphocytic lymphoma (SLL).

[0309] In the twenty - seventh embodiment (Embodiment E27), the invention includes a method of treating cancer in a human patient in need thereof, comprising subcutaneously administering to the patient a dose of about 600 mg to about 1000 mg of an anti - PD - 1 antibody (e.g., pembrolizumab) or an antigen - binding fragment thereof, about once every six weeks, wherein the patient has a tumor with a high mutational burden. In a sub - embodiment of Embodiment E27, the tumor is a solid tumor. In a particular embodiment, the patient is an adult human patient. In a particular embodiment, the patient is a pediatric patient.

[0310] In a sub - embodiment of Embodiment E27, the high mutational burden is at least about 10 mutations per megabase of the genome examined. In other embodiments, the high mutational burden is at least about 11 mutations per megabase of the genome examined, at least about 12 mutations per megabase of the genome examined, or at least about 13 mutations per megabase of the genome examined.

[0311] In the twenty - eighth embodiment (Embodiment E28), the invention includes a method of treating esophageal cancer in a human patient in need thereof, comprising subcutaneously administering to the patient a dose of about 600 mg to about 1000 mg of an anti - PD - 1 antibody (e.g., pembrolizumab) or an antigen - binding fragment thereof, about once every six weeks.

[0312] In a sub - embodiment of Embodiment E28, the patient has undergone one prior line of standard therapy before receiving the anti - PD - 1 antibody or an antigen - binding fragment thereof. In a further embodiment, the patient has undergone one or more lines of standard therapy before receiving the anti - PD - 1 antibody or an antigen - binding fragment thereof. In another embodiment, the patient has undergone two or more lines of standard therapy before receiving the anti - PD - 1 antibody or an antigen - binding fragment thereof. In a particular embodiment, the standard therapy includes one or more of the following: paclitaxel, docetaxel, or irinotecan.

[0313] In a sub - embodiment of Embodiment E28, the patient has advanced or metastatic adenocarcinoma or squamous cell carcinoma of the esophagus.

[0314] In a sub - embodiment of Embodiment E28, the patient has advanced or metastatic Siewert type I adenocarcinoma of the esophagogastric junction.

[0315] In a sub - embodiment of Embodiment E28, the patient's tumor expresses PD - L1 (Combined Positive Score [CPS] ≥ 10).

[0316] In the twenty - ninth embodiment (Embodiment E29), the present invention includes a method for treating high - risk non - muscle - invasive bladder cancer (NMIBC) in a patient in need thereof, which comprises subcutaneously administering to the patient a dose of about 600 mg to about 1000 mg of an anti - PD - 1 antibody (e.g., pembrolizumab) or an antigen - binding fragment thereof approximately once every six weeks. In certain embodiments, the patient has NMIBC and carcinoma in situ (CIS) or CIS plus papillary disease.

[0317] In a sub - embodiment of Embodiment E29, the patient has previously been treated with standard therapy prior to treatment with the anti - PD - 1 antibody or an antigen - binding fragment thereof. In certain embodiments, the previous therapy is Bacillus Calmette - Guérin (BCG) therapy. In certain embodiments, the patient is non - responsive to BCG therapy. In certain embodiments, the patient is ineligible for radical cystectomy or elects not to undergo radical cystectomy.

[0318] In the thirtieth embodiment (Embodiment E30), the present invention includes a method for treating cutaneous squamous cell carcinoma (cSCC) in a human patient in need thereof, which comprises subcutaneously administering to the patient a dose of about 600 mg to about 1000 mg of an anti - PD - 1 antibody (e.g., pembrolizumab) or an antigen - binding fragment thereof approximately once every six weeks. In certain embodiments, the cutaneous squamous cell carcinoma is incurable by surgery or radiotherapy.

[0319] In the thirty - first embodiment (Embodiment E31), the present invention includes a method for treating endometrial cancer in a human patient in need thereof, which comprises subcutaneously administering to the patient a dose of about 600 mg to about 1000 mg of an anti - PD - 1 antibody (e.g., pembrolizumab) or an antigen - binding fragment thereof approximately once every six weeks.

[0320] In some sub - embodiments of Embodiment E31, the endometrial cancer is advanced endometrial cancer that is non - MSI - H or mismatch repair - deficient (dMMR). In certain embodiments, the patient has disease progression after previous systemic therapy.

[0321] In some sub - embodiments of embodiment E31, the endometrial cancer is advanced endometrial cancer that is MSI - H or dMMR, as determined by an FDA - approved test, where the patient has had disease progression following prior systemic therapy in any case. In a particular embodiment, the patient is not suitable for curative surgery or radiotherapy.

[0322] In the thirty - second embodiment (embodiment E32), the invention includes a method of treating cervical cancer in a human patient in need thereof, comprising subcutaneously administering to the patient a dose of about 600 mg to about 1000 mg of an anti - PD - 1 antibody (e.g., pembrolizumab) or an antigen - binding fragment thereof approximately once every six weeks. In one embodiment, the cervical cancer is recurrent or metastatic cervical cancer and the patient has had disease progression during or after chemotherapy.

[0323] In a sub - embodiment of embodiment E32, the method further comprises treating the patient with chemotherapy with or without bevacizumab. In a particular embodiment, the cervical cancer is persistent, recurrent or metastatic cervical cancer and the patient's tumor expresses PD - L1 (CPS > 1). In one embodiment, the chemotherapy is paclitaxel 175 mg / m 2 and cisplatin 50 mg / m 2 or paclitaxel 175 mg / m 2 and carboplatin AUC 5 mg / mL / min, and is administered on day 1 of every three weeks. In one embodiment, the chemotherapy with bevacizumab is paclitaxel 175 mg / m 2 and cisplatin 50 mg / m 2 or paclitaxel 175 mg / m 2 and carboplatin AUC 5 mg / mL / min and bevacizumab 15 mg / kg, and is administered on day 1 of every three weeks.

[0324] In a sub - embodiment of embodiment E32, the cervical cancer is recurrent or metastatic cervical cancer that has had disease progression during or after chemotherapy and the patient's tumor expresses PD - L1 (CPS > 1).

[0325] In the thirty - third embodiment (embodiment E33), the invention includes a method of treating stage IB, II or IIIA non - small cell lung cancer (NSCLC) in a human patient in need thereof, comprising subcutaneously administering to the patient a dose of about 600 mg to about 1000 mg of an anti - PD - 1 antibody (e.g., pembrolizumab) or an antigen - binding fragment thereof approximately once every six weeks for adjuvant treatment after resection.

[0326] In a sub - embodiment (E33 - A) of embodiment E33, the patient has previously received platinum - based chemotherapy.

[0327] In any method or use of the invention described herein (including embodiments E1 - E33), the anti - PD - 1 antibody or its antigen - binding fragment is any antibody or antigen - binding fragment described in Section II, "PD - 1 Antibodies and Antigen - Binding Fragments Useful in the Invention" of the specific embodiments of the invention herein. In a particular embodiment, the anti - PD - 1 antibody is pembrolizumab or its antigen - binding fragment, or an antibody that cross - competes with pembrolizumab for binding to human PD - 1. In a particular embodiment, the anti - PD - 1 antibody is a pembrolizumab variant.

[0328] In any method or use of the invention described herein (including embodiments E1 - E33), the anti - PD - 1 antibody or its antigen - binding fragment can be co - administered or co - formulated with the hyaluronidase described in Section III. In one embodiment, the anti - PD - 1 antibody or its antigen - binding fragment is co - administered with human hyaluronidase. In another embodiment, the anti - PD - 1 antibody or its antigen - binding fragment is co - formulated with human hyaluronidase. In a particular embodiment, the human hyaluronidase is rHuPH20. In a particular embodiment, the human hyaluronidase is PH20 variant 2.

[0329] In any method or use of the invention described herein (including embodiments E1 - E33), the anti - PD - 1 antibody or its antigen - binding fragment is administered to a patient at a dose of about 600 mg to about 1000 mg every six weeks. In a further embodiment, the anti - PD - 1 antibody or its antigen - binding fragment is administered to a patient at a dose of about 600 mg to about 950 mg every six weeks. In a further embodiment, the anti - PD - 1 antibody or its antigen - binding fragment is administered to a patient at a dose of about 600 mg to about 900 mg every six weeks. In a further embodiment, the anti - PD - 1 antibody or its antigen - binding fragment is administered to a patient at a dose of about 650 mg to about 800 mg every six weeks. In a further embodiment, the anti - PD - 1 antibody or its antigen - binding fragment is administered to a patient at a dose of about 660 mg to about 800 mg every six weeks. In a further embodiment, the anti - PD - 1 antibody or its antigen - binding fragment is administered to a patient at a dose of about 700 mg to about 800 mg every six weeks. In a further embodiment, the anti - PD - 1 antibody or its antigen - binding fragment is administered to a patient at a dose of about 760 mg to about 800 mg every six weeks. In a further embodiment, the anti - PD - 1 antibody or its antigen - binding fragment is administered to a patient at a dose of about 760 mg to about 790 mg every six weeks.

[0330] In any method or use of the invention described herein (including embodiments E1-E33), the anti-PD-1 antibody or antigen-binding fragment is administered at a dose of 600 mg to 1000 mg every six weeks. In a further embodiment, the anti-PD-1 antibody or antigen-binding fragment is administered to a patient at a dose of 600 mg to 950 mg every six weeks. In a further embodiment, the anti-PD-1 antibody or antigen-binding fragment is administered at a dose of from 600 mg to 900 mg every six weeks. In a further embodiment, the anti-PD-1 antibody or its antigen-binding fragment is administered to a patient at a dose of 650 mg to 800 mg every six weeks. In a further embodiment, the anti-PD-1 antibody or its antigen-binding fragment is administered to a patient at a dose of from 660 mg to 800 mg every six weeks. In a further embodiment, the anti-PD-1 antibody or antigen-binding fragment is administered at a dose of 700 mg to 800 mg every six weeks. In a further embodiment, the anti-PD-1 antibody or antigen-binding fragment is administered at a dose of 760 mg to 800 mg every six weeks. In a further embodiment, the anti-PD-1 antibody or antigen-binding fragment is administered at a dose of 760 mg to 790 mg every six weeks.

[0331] In any method or use of the invention described herein (including embodiments E1-E33), the anti-PD-1 antibody or its antigen-binding fragment is administered at a dose of 760 mg every six weeks. In one embodiment, the anti-PD-1 antibody or its antigen-binding fragment is administered at a dose of 770 mg every six weeks. In another embodiment, the anti-PD-1 antibody or its antigen-binding fragment is administered at a dose of 780 mg every six weeks. In another embodiment, the anti-PD-1 antibody or its antigen-binding fragment is administered at a dose of 790 mg every six weeks. In another embodiment, the anti-PD-1 antibody or its antigen-binding fragment is administered at a dose of 800 mg every six weeks. In another embodiment, the anti-PD-1 antibody or its antigen-binding fragment is administered at a dose of 810 mg every six weeks. In another embodiment, the anti-PD-1 antibody or its antigen-binding fragment is administered at a dose of 650 mg every six weeks. In another embodiment, the anti-PD-1 antibody or its antigen-binding fragment is administered at a dose of 750 mg every six weeks.

[0332] In any method or use of the invention described herein (including embodiments E1 - E33), the anti - PD - 1 antibody or an antigen - binding fragment thereof is administered subcutaneously to a patient at a dose of about 300 mg to about 500 mg every about three weeks in combination with human hyaluronidase (including those described in Section III). In any method or use of the invention described herein (including embodiments E1 - E33), the anti - PD - 1 antibody or an antigen - binding fragment thereof is administered subcutaneously to a patient at a dose of about 300 mg to about 500 mg every about three weeks in combination with PH20 variant 2. In any method or use of the invention described herein (including embodiments E1 - E33), the anti - PD - 1 antibody or an antigen - binding fragment thereof is administered subcutaneously to a patient at a dose of about 300 mg to about 500 mg every about three weeks in combination with rHuPH20. In certain embodiments, the anti - PD - 1 antibody or an antigen - binding fragment thereof is administered subcutaneously to a patient every three weeks, every three weeks plus or minus 5 days, plus or minus 4 days, plus or minus 3 days, plus or minus 2 days, or plus or minus 1 day.

[0333] In a further embodiment, the anti - PD - 1 antibody or antigen - binding fragment is administered in combination with human hyaluronidase at a dose of about 300 mg to about 500 mg every three weeks. In a further embodiment, the anti - PD - 1 antibody or antigen - binding fragment is administered in combination with human hyaluronidase at a dose of about 350 mg to about 450 mg every three weeks. In a further embodiment, the anti - PD - 1 antibody or antigen - binding fragment is administered in combination with human hyaluronidase at a dose of about 360 mg to about 420 mg every three weeks. In a further embodiment, the anti - PD - 1 antibody or antigen - binding fragment is administered in combination with human hyaluronidase at a dose of about 380 mg to about 120 mg every three weeks. In a further embodiment, the anti - PD - 1 antibody or antigen - binding fragment is administered in combination with human hyaluronidase at a dose of 380 mg every three weeks. In a further embodiment, the anti - PD - 1 antibody or antigen - binding fragment is administered in combination with human hyaluronidase at a dose of 350 mg every three weeks. In another embodiment, the anti - PD - 1 antibody or antigen - binding fragment is administered in combination with human hyaluronidase at a dose of 390 mg every three weeks. In another embodiment, the anti - PD - 1 antibody or antigen - binding fragment is administered in combination with human hyaluronidase at a dose of 395 mg every three weeks. In another embodiment, the anti - PD - 1 antibody or antigen - binding fragment is administered in combination with human hyaluronidase at a dose of 325 mg every three weeks. In another embodiment, the anti - PD - 1 antibody or antigen - binding fragment is administered in combination with human hyaluronidase at a dose of 400 mg every three weeks.

[0334] In a further embodiment, the anti-PD-1 antibody or antigen-binding fragment is administered with human hyaluronidase at a dose of 300 mg to 500 mg every three weeks. In a further embodiment, the anti-PD-1 antibody or antigen-binding fragment is administered with human hyaluronidase at a dose of 350 mg to 450 mg every three weeks. In a further embodiment, the anti-PD-1 antibody or antigen-binding fragment is administered with human hyaluronidase at a dose of 360 mg to 420 mg every three weeks. In a further embodiment, the anti-PD-1 antibody or antigen-binding fragment is administered with human hyaluronidase at a dose ranging from 380 mg to 410 mg every three weeks.

[0335] In any method or use of the invention described herein (including embodiments E1-E33), the anti-PD-1 antibody or its antigen-binding fragment is co-administered with the human hyaluronidase PH20 variant or fragment as defined herein. In another embodiment, the anti-PD-1 antibody or its antigen-binding fragment is co-formulated with PH20 variant 2 in a composition. For example, US20220089739, the content of which is incorporated herein by reference, describes the preparation of a liquid composition comprising pembrolizumab and a PH20 variant.

[0336] In one embodiment, the composition comprises 130 mg / ml of the anti-PD-1 antibody or its antigen-binding fragment. In other embodiments, the composition comprises 165 mg / ml of the anti-PD-1 antibody or its antigen-binding fragment.

[0337] In a further embodiment, the composition further comprises L-methionine. In a particular embodiment, L-methionine is present at a concentration of about 10 mM.

[0338] In a further embodiment, the composition further comprises a histidine buffer at about pH 5.0 to pH 6.0. In a particular embodiment, histidine is present at a concentration of about 10 mM.

[0339] In a further embodiment, the composition further comprises sucrose. In a particular embodiment, sucrose is present at a concentration of about 70 mg / mL. In a particular embodiment, sucrose is present at a concentration of 7% (w / v).

[0340] In a further embodiment of the invention, the composition further comprises polysorbate 80. In a particular embodiment, polysorbate 80 is present at a concentration of about 0.2 mg / mL. In a particular embodiment, polysorbate 80 is present at a concentration of 0.02% (w / v).

[0341] In certain embodiments, the composition comprises 10 mM L-methionine, 10 mM histidine, pH 5.5, 7% sucrose, 0.02% polysorbate 80, and 130 mg / mL anti-PD-1 antibody or antigen-binding fragment thereof.

[0342] In certain embodiments, the composition comprises 10 mM L-methionine, 10 mM histidine, pH 5.5, 7% sucrose, 0.02% polysorbate 80, and 165 mg / mL anti-PD-1 antibody or antigen-binding fragment thereof.

[0343] In certain embodiments of the methods or uses described herein, the anti-PD-1 antibody or antigen-binding fragment thereof is administered subcutaneously in one or more injections. In certain embodiments, the anti-PD-1 antibody or antigen-binding fragment thereof is administered in 2 injections.

[0344] In one embodiment, 790 mg of anti-PD-1 antibody or antigen-binding fragment thereof is administered subcutaneously in one injection as a composition comprising 130 mg / mL. In one embodiment, 790 mg of anti-PD-1 antibody or antigen-binding fragment thereof is administered subcutaneously in two injections.

[0345] In one embodiment, 790 mg of anti-PD-1 antibody or antigen-binding fragment thereof is administered subcutaneously in one injection as a composition comprising 165 mg / mL. In one embodiment, 790 mg of anti-PD-1 antibody or antigen-binding fragment thereof is administered subcutaneously in two injections as a composition comprising 165 mg / mL. In a further embodiment, 4.8 mL of a composition comprising 165 mg / mL anti-PD-1 antibody or antigen-binding fragment thereof is administered subcutaneously in one injection.

[0346] In one embodiment, 395 mg of anti-PD-1 antibody or antigen-binding fragment thereof is administered subcutaneously in one injection as a composition comprising 130 mg / mL. In one embodiment, 395 mg of anti-PD-1 antibody or antigen-binding fragment thereof is administered subcutaneously in two injections.

[0347] In one embodiment, 395 mg of anti-PD-1 antibody or antigen-binding fragment thereof is administered subcutaneously in one injection as a composition comprising 165 mg / mL. In one embodiment, 395 mg of anti-PD-1 antibody or antigen-binding fragment thereof is administered subcutaneously in two injections as a composition comprising 165 mg / mL. In a further embodiment, 2.4 mL of a composition comprising 165 mg / mL anti-PD-1 antibody or antigen-binding fragment thereof is administered subcutaneously in one injection. In a further embodiment, 1.2 mL of a composition comprising 165 mg / mL anti-PD-1 antibody or antigen-binding fragment thereof is administered subcutaneously in two separate injections.

[0348] In any method and use of the present invention, the anti-PD-1 antibody or antigen-binding fragment thereof at the doses described herein is co-formulated or co-administered with a dose of human hyaluronidase from about 700 units to about 50,000 units. In one embodiment, the dose is from about 2,000 units to about 20,000 units of human hyaluronidase. In one embodiment, the dose is from about 5,000 units to about 15,000 units of human hyaluronidase. In one embodiment, from about 10,000 units to about 30,000 units of human hyaluronidase is co-formulated or co-administered with a dose of the anti-PD-1 antibody or antigen-binding fragment thereof. In one embodiment, the dose is from about 7,000 units to about 13,000 units of human hyaluronidase. In one embodiment, the dose is from about 8,000 units to about 10,000 units of human hyaluronidase. In one embodiment, the dose is from about 4,000 units to about 6,000 units of human hyaluronidase. In one embodiment, the dose is 4,800 units of human hyaluronidase. In one embodiment, the dose is 9,600 units of human hyaluronidase.

[0349] In one embodiment, human hyaluronidase is co-formulated or co-administered with the anti-PD-1 antibody or antigen-binding fragment thereof at a ratio of about 12.15 units: 1 mg. In one embodiment, human hyaluronidase is co-formulated or co-administered with the anti-PD-1 antibody or antigen-binding fragment thereof at a ratio of about 12 units: 1 mg. In one embodiment, human hyaluronidase is co-formulated or co-administered with the anti-PD-1 antibody or antigen-binding fragment thereof at a ratio of about 10 to 14 units: 1 mg. In one embodiment, human hyaluronidase is co-formulated or co-administered with the anti-PD-1 antibody or antigen-binding fragment thereof at a ratio of about 8 to 18 units: 1 mg. In one embodiment, human hyaluronidase is co-formulated or co-administered with the anti-PD-1 antibody or antigen-binding fragment thereof at a ratio of about 6 to 25 units: 1 mg. In one embodiment, human hyaluronidase is co-formulated or co-administered with the anti-PD-1 antibody or antigen-binding fragment thereof at a ratio of about 3 to 36 units: 1 mg.

[0350] In any of the methods or uses described herein, including embodiments E1 - E33 and their sub - embodiments, the method may further comprise administering one or more "other therapeutic agents" (as used herein, "other therapeutic agents" refers to other agents relative to the anti - PD - 1 antibody or its antigen - binding fragment). The other therapeutic agents can be, for example, chemotherapeutic agents, biotherapeutic agents (including but not limited to antibodies against CTLA4, TIGIT, VEGF, EGFR, Her2 / neu, VEGF receptors, other growth factor receptors, CD20, CD40, CD - 40L, OX - 40, 4 - 1BB, and ICOS), immunogenic agents (e.g., attenuated cancer cells, tumor antigens, antigen - presenting cells such as dendritic cells pulsed with tumor - derived antigens or nucleic acids, immunostimulatory cytokines (e.g., IL - 2, IFNα2, GM - CSF), and cells transfected with genes encoding immunostimulatory cytokines (such as but not limited to GM - CSF)).

[0351] As shown above, in certain embodiments of the methods or uses of the present invention, the method further comprises administering other therapeutic agents. In certain embodiments, the other therapeutic agents are an anti - CTLA4 antibody or its antigen - binding fragment, an anti - LAG3 antibody or its antigen - binding fragment, an anti - GITR antibody or its antigen - binding fragment, and an anti - TIGIT antibody or its antigen - binding fragment, an anti - CD27 antibody or its antigen - binding fragment, an anti - ILT3 antibody or its antigen - binding fragment, or an anti - ILT4 antibody or its antigen - binding fragment. In one embodiment, the other therapeutic agent is a Newcastle disease virus vector expressing IL - 12. In a further embodiment, the other therapeutic agent is dinaciclib. In another embodiment, the other therapeutic agent is navarixin. In a further embodiment, the other therapeutic agent is vicriviroc.

[0352] In a further embodiment, the other therapeutic agent is an oncolytic virus. In one embodiment, the other therapeutic agent is Coxsackievirus or CVA21. In one embodiment, the other therapeutic agent is CAVATAK TM 。

[0353] In yet another embodiment, the other therapeutic agent is a STING agonist. In a further embodiment, the other therapeutic agent is an IL-27 antagonist. In one embodiment, the other therapeutic agent is a PARP inhibitor. In one embodiment, the other therapeutic agent is a multi-kinase inhibitor. In one embodiment, the other therapeutic agent is a MEK inhibitor. In one embodiment, the other therapeutic agent is a 4-1BB agonist. In one embodiment, the other therapeutic agent is nemtabrutinib. In another embodiment, the other therapeutic agent is belzutifan.

[0354] Examples of chemotherapeutic agents include alkylating agents such as thiotepa and cyclophosphamide; alkyl sulfonates such as busulfan, improsulfan and piposulfan; aziridines such as benzodopa, carboquone, meturedopa and uredepa; ethyleneimines and methylamelamines including altretamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramide and trimethylolomelamine; polyacetylenes (especially bullatacin and bullatacinone); camptothecin (including the synthetic analogue topotecan); bryostatin; calicheamicin; CC-1065 (including its synthetic analogues adozelesin, carzelesin and bizelesin); cryptophycin (especially cryptophycin 1 and cryptophycin 8); dolastatin; duocarmycin (including the synthetic analogues KW-2189 and CB1-TM1); eleutherobin; pancratistatin; sarcodictyin; spongistatin; nitrogen mustards such as chlorambucil, chlornaphazine, chlorophosphamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, novembichin, phenesterine, prednimustine, trophosphamide, uracil mustard; nitrosureas such as carmustine, chlorozotocin, fotemustine, lomustine, nimustine and ranimustine; antibiotics such as the enediyne antibiotics (e.g., calicheamicin, especially calicheamicin γ and calicheamicin ω (see, e.g., Agnew, Chem Intl. Ed. Engl, 33:183-186 (1994)); dynemicin, including dynemicin A; bisphosphonates such as clodronic acid; esperamicin;and neocarzinostatin chromophore and related chromoprotein enediyne antibiotic chromophores, aclacinomysin, actinomycin, anthramycin, azaserine, bleomycin, actinomycin C, carzinophilin, chromomycin, actinomycin D, daunomycin, detorubicin, 6-diazo-5-oxo-L-norleucine, doxorubicin (including morpholino-doxorubicin, cyanomorpholino-doxorubicin, 2-pyrrolinyl-doxorubicin and deoxydoxorubicin), epirubicin, esorubicin, idarubicin, marcellomycin, mitomycin (such as mitomycin C), mycophenolic acid, nogalamycin, olivomycin, peplomycin, potfiromycin, puromycin, quelamycin, rodorubicin, streptonigrin, streptozocin, tubercidin, bestatin, zinostatin, zorubicin; antimetabolites, such as methotrexate and 5-fluorouracil (5-FU); folic acid analogues, such as denopterin, methotrexate, pteropterin, trimetrexate; purine analogues, such as fludarabine, 6-mercaptopurine, thioguanine, thioinosine; pyrimidine analogues, such as ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, didoxuridine, floxuridine, enocitabine, fluorouracil; androgens, such as calusterone, drostanolone propionate, epitiostanol, methylandrostane, testolactone; antiadrenal agents, such as aminoglutethimide, mitotane, trilostane; folic acid supplements, such as leucovorin; acetylcysteine; aldophosphamide glycoside; aminolevulinic acid; eniluracil; ametantrone; bexarotene; bisantrene; edatrexate; defofamine; colchicine amide; diaziquone; eflornithine; elisidepsin; epothilone; etoglucid; gallium nitrate; hydroxyurea; lentinan; lonidamine; maytansinoids, such as maytansine and ansamitocin; mitoguazone; mitoxantrone; mopidanmol; nitracrine; pentostatin; phenamet; pirarubicin; losoxantrone; podophyllinic acid; 2-ethylhydrazide; procarbazine; razoxane; rhizoxin; sizofiran; spirogermanium; tenuazonic acid; triaziquone;2,2′,2″-Trichloroethylamine; trichothecene (especially T-2 toxin, verracurin A, roridin A, and anguidine); urethane; vindesine; dacarbazine; mannomustine; dibromomannitol; mitolactol; pipobroman; gacytosine; cytarabine (“Ara-C”); cyclophosphamide; thiotepa; taxane, such as paclitaxel and docetaxel; chlorambucil; gemcitabine; 6-thioguanine; mercaptopurine; methotrexate; platinum analogs, such as cisplatin and carboplatin; vinblastine; platinum; etoposide (VP-16); ifosfamide; mitoxantrone; vincristine; vinorelbine; novantrone; teniposide; edatrexate; daunomycin; aminopterin; capecitabine; ibandronate; CPT-11; topoisomerase inhibitor RFS2000; difluoromethylornithine (DMFO); retinoid, such as retinoic acid; capecitabine; and a pharmaceutically acceptable salt, acid, or derivative of any of the foregoing. Also included are antihormonal agents for modulating or inhibiting the action of hormones on tumors, such as antiestrogens and selective estrogen receptor modulators (SERM), including, for example, tamoxifen, raloxifene, droloxifene, 4-hydroxytamoxifen, trioxifene, clomoxifene, LY117018, onapristone, and toremifene (Fareston); aromatase inhibitors that inhibit aromatase, which regulates estrogen production in the adrenal gland, such as, for example, 4(5)-imidazole, aminoglutethimide, megestrol acetate, exemestane, formestane, fadrozole, vorozole, letrozole, and anastrozole; and antiandrogens, such as flutamide, nilutamide, bicalutamide, leuprolide, and goserelin; and a pharmaceutically acceptable salt, acid, or derivative of any of the foregoing.;

[0355] In certain embodiments that include the step of administering other therapeutic agents (i.e., other than an anti-PD-1 antibody (e.g., pembrolizumab) or an antigen-binding fragment thereof), the other therapeutic agents in the combination therapy can be administered using the same dosing regimen (dose, frequency, and duration of treatment) as is typically used when the agent is used as a single therapy for treating the same cancer. In other embodiments, the total amount of the other therapeutic agent received by the patient in the combination therapy is lower than the total amount of the agent as a single therapy, e.g., a smaller dose, a lower frequency dose, and / or a shorter treatment duration.

[0356] The appropriate dosage is determined by the clinician, for example, using parameters or factors known or suspected to affect or predicted to affect the treatment in the art. The appropriate dosage of the medicament (“therapeutically effective amount”) will depend on, for example, the condition to be treated, the severity and duration of the condition, whether the medicament is administered for prophylactic or therapeutic purposes, previous therapies, the patient's clinical history and response to the medicament, the type of medicament used, and the judgment of the attending physician. Generally, the dosage starts at an amount slightly below the optimal dose and is then increased in smaller increments until the desired or optimal effect is achieved relative to any negative side effects.

[0357] The other therapeutic agents in the combination therapy can be administered orally, intratumorally, or parenterally (including intravenous, intramuscular, intraperitoneal, subcutaneous, rectal, topical, and transdermal administration routes). For example, the combination therapy can comprise an anti-PD-1 antibody or an antigen-binding fragment thereof and an anti-CTLA antibody or an antigen-binding fragment thereof, both of which can be administered intravenously or subcutaneously, and a chemotherapeutic agent, which can be administered orally.

[0358] In certain embodiments, the combination therapy of the present invention can be administered before or after surgical removal of the tumor and can be used before, during, or after radiotherapy. The combination therapy of the present invention can also be used when the patient's tumor is inoperable.

[0359] In certain embodiments, the combination therapy of the present invention is administered to a patient who has not been previously treated with a biotherapeutic agent or a chemotherapeutic agent, i.e., an initial treatment. In other embodiments, the combination therapy is administered to a patient who has not achieved a sustained response after previous therapy with a biotherapeutic agent or a chemotherapeutic agent, i.e., a patient who has undergone treatment.

[0360] The combination therapy of the present invention can be used to treat tumors large enough to be detected by palpation or by imaging techniques well known in the art, such as MRI, ultrasound, or CAT scan. In certain embodiments, the combination therapy of the present invention is used to treat advanced tumors having a diameter of at least about 200 mm 3 、300 mm 3 、400 mm 3 、500 mm 3 、750 mm 3 or up to 1000 mm 3 in diameter.

[0361] In certain embodiments, the combination therapy of the invention is administered to a human patient having cancer expressing PD-L1. In certain embodiments, PD-L1 expression is detected on FFPE or frozen tissue sections of a tumor sample removed from the patient using a diagnostic anti-human PD-L1 antibody or antigen-binding fragment thereof in an IHC assay. The patient's physician may direct a diagnostic test to determine PD-L1 expression in a tumor tissue sample removed from the patient prior to initiating treatment with an anti-PD-1 antibody or antigen-binding fragment thereof, but it is contemplated that the physician may direct a first or subsequent diagnostic test at any time after initiating treatment, such as, for example, after completion of a treatment cycle.

[0362] V. Kits and Compositions

[0363] The invention also relates to a pharmaceutical composition for subcutaneous injection comprising an anti-PD-1 antibody or antigen-binding fragment thereof in a dose of about 600 mg to about 1000 mg and about 700 units to about 50,000 units of human hyaluronidase. The invention also relates to a pharmaceutical composition for subcutaneous injection comprising an anti-PD-1 antibody or antigen-binding fragment thereof in a dose of about 600 mg to about 1000 mg and about 7290 units to about 12,150 units of human hyaluronidase. In one embodiment, the pharmaceutical composition for subcutaneous injection comprises an anti-PD-1 antibody or antigen-binding fragment thereof in a dose of about 700 mg to about 800 mg and about 700 units to about 50,000 units of human hyaluronidase. In one embodiment, the pharmaceutical composition for subcutaneous injection comprises an anti-PD-1 antibody or antigen-binding fragment thereof in a dose of about 700 mg to about 800 mg and about 8505 units to about 9720 units of human hyaluronidase. In one embodiment, the pharmaceutical composition for subcutaneous injection comprises 790 mg of an anti-PD-1 antibody or antigen-binding fragment thereof and 9600 units of human hyaluronidase.

[0364] The invention also relates to a pharmaceutical composition for subcutaneous injection comprising an anti-PD-1 antibody or antigen-binding fragment thereof in a dose of about 600 mg to about 1000 mg. In one embodiment, the pharmaceutical composition for subcutaneous injection comprises an anti-PD-1 antibody or antigen-binding fragment thereof in a dose of about 700 mg to about 800 mg. In one embodiment, the pharmaceutical composition for subcutaneous injection comprises an anti-PD-1 antibody or antigen-binding fragment thereof in a dose of about 760 mg to about 790 mg. In one embodiment, the pharmaceutical composition for subcutaneous injection comprises 790 mg of an anti-PD-1 antibody or antigen-binding fragment thereof.

[0365] The present invention also relates to a pharmaceutical composition for subcutaneous injection, which comprises an anti-PD-1 antibody or an antigen-binding fragment thereof in a dose of about 300 mg to about 500 mg and about 700 units to about 50,000 units of human hyaluronidase. The present invention also relates to a pharmaceutical composition for subcutaneous injection, which comprises an anti-PD-1 antibody or an antigen-binding fragment thereof in a dose of about 300 mg to about 500 mg and about 3,645 units to about 6,075 units of human hyaluronidase. In one embodiment, the pharmaceutical composition for subcutaneous injection comprises an anti-PD-1 antibody or an antigen-binding fragment thereof in a dose of about 350 mg to about 400 mg and about 700 units to about 50,000 units of human hyaluronidase. In one embodiment, the pharmaceutical composition for subcutaneous injection comprises an anti-PD-1 antibody or an antigen-binding fragment thereof in a dose of about 350 mg to about 400 mg and about 4,252 units to 4,860 units of human hyaluronidase. In one embodiment, the pharmaceutical composition for subcutaneous injection comprises an anti-PD-1 antibody in a dose of 395 mg or an antigen-binding fragment thereof and 4,800 units of human hyaluronidase. In one embodiment, the pharmaceutical composition for subcutaneous injection comprises 395 mg of pembrolizumab and about 33 μg of human hyaluronidase of SEQ ID NO: 18. In another embodiment, the pharmaceutical composition for subcutaneous injection comprises 790 mg of pembrolizumab and about 66 μg of human hyaluronidase of SEQ ID NO: 18. In one embodiment, the pharmaceutical composition for subcutaneous injection comprises an anti-PD-1 antibody in a dose of 395 mg and about 33 μg of human hyaluronidase of SEQ ID NO: 18. In another embodiment, the pharmaceutical composition for subcutaneous injection comprises an anti-PD-1 antibody in a dose of 790 mg and about 66 μg of human hyaluronidase of SEQ ID NO: 18. In one embodiment of the foregoing embodiments, the anti-PD-1 antibody is a monoclonal antibody, which comprises a heavy chain consisting of the amino acid sequence shown in SEQ ID NO: 11 and a light chain consisting of the amino acid sequence shown in SEQ ID NO: 5.

[0366] In any of the methods, uses, and compositions described herein, from about 700 units to about 50,000 units of human hyaluronidase are co-formulated with a dose of an anti-PD-1 antibody or an antigen-binding fragment thereof. In one embodiment, from about 3,000 units to about 15,000 units of human hyaluronidase are co-formulated with a dose of an anti-PD-1 antibody or an antigen-binding fragment thereof. In one embodiment, from about 3,000 units to about 7,000 units of human hyaluronidase are co-formulated with a dose of an anti-PD-1 antibody or an antigen-binding fragment thereof. In one embodiment, from about 7,000 units to about 14,000 units of human hyaluronidase are co-formulated with a dose of an anti-PD-1 antibody or an antigen-binding fragment thereof. In one embodiment, from about 10,000 units to about 30,000 units of human hyaluronidase are co-formulated with a dose of an anti-PD-1 antibody or an antigen-binding fragment thereof. In one embodiment, from about 8,000 units to about 10,000 units of human hyaluronidase are co-formulated with a dose of an anti-PD-1 antibody or an antigen-binding fragment thereof. In one embodiment, from about 4,000 units to about 5,000 units of human hyaluronidase are co-formulated with a dose of an anti-PD-1 antibody or an antigen-binding fragment thereof. In one embodiment, about 9,600 units of human hyaluronidase are co-formulated with a dose of an anti-PD-1 antibody or an antigen-binding fragment thereof. In one embodiment, about 4,800 units of human hyaluronidase are co-formulated with a dose of an anti-PD-1 antibody or an antigen-binding fragment thereof.

[0367] In one embodiment, human hyaluronidase and an anti-PD-1 antibody or an antigen-binding fragment thereof are co-formulated at a ratio of about 12.15 units: 1 mg. In one embodiment, human hyaluronidase and an anti-PD-1 antibody or an antigen-binding fragment thereof are co-formulated at a ratio of about 12 units: 1 mg. In one embodiment, human hyaluronidase and an anti-PD-1 antibody or an antigen-binding fragment thereof are co-formulated at a ratio of about 10 to 14 units: 1 mg. In one embodiment, human hyaluronidase and an anti-PD-1 antibody or an antigen-binding fragment thereof are co-formulated at a ratio of about 8 to 18 units: 1 mg. In one embodiment, human hyaluronidase and an anti-PD-1 antibody or an antigen-binding fragment thereof are co-formulated at a ratio of about 6 to 25 units: 1 mg. In one embodiment, human hyaluronidase and an anti-PD-1 antibody or an antigen-binding fragment thereof are co-formulated at a ratio of about 3 to 36 units: 1 mg.

[0368] In the foregoing embodiments, the pharmaceutical composition may be contained in a vial, an in vivo device, or one or more prefilled syringes. Any of the pharmaceutical compositions described herein may be used in the methods or uses described in Section IV.

[0369] The present invention also relates to a kit for treating a patient suffering from cancer, the kit comprising: (a) a composition for subcutaneous injection, which comprises an anti-PD-1 antibody or an antigen-binding fragment thereof in a dose of about 600 mg to about 1000 mg or about 300 mg to about 500 mg (or the dose or composition described in Section IV or V), and optionally human hyaluronidase (in any amount described in Sections III, IV or V), or any pharmaceutical composition in the foregoing embodiments of this section; and (b) instructions for using the composition or pharmaceutical composition of (a) in any method or use for treating cancer described herein.

[0370] The kit of the present invention may provide an anti-PD-1 antibody or an antigen-binding fragment thereof and optionally human hyaluronidase in a container, together with package inserts. The container contains a dose of a composition comprising at least about 600 mg to about 1000 mg, or about 300 mg to about 500 mg, of an anti-PD-1 antibody or an antigen-binding fragment thereof, and optionally about 70 units to about 50,000 units of human hyaluronidase, as well as a package insert or label comprising instructions for treating a patient suffering from cancer with the composition. The container may be of any shape and / or material (e.g., plastic or glass). For example, the container may be a vial, syringe or bottle. The kit may further comprise other materials that can be used to administer the agent, such as in vivo devices, needles and syringes. In a particular embodiment of the kit, the instructions state that the agent is intended for treating a patient as described in any one of Embodiments E1-E33 of the section headed "Methods and Uses of the Present Invention" in Section IV above.

[0371] In one embodiment, the composition comprises 130 mg / ml of an anti-PD-1 antibody or an antigen-binding fragment thereof. In other embodiments, the composition comprises 165 mg / ml of an anti-PD-1 antibody or an antigen-binding fragment thereof.

[0372] In a further embodiment, the composition further comprises L-methionine. In a particular embodiment, L-methionine is present at a concentration of about 10 nM.

[0373] In a further embodiment, the composition further comprises a histidine buffer at about pH 5.0 to pH 6.0. In a particular embodiment, histidine is present at a concentration of about 10 mM.

[0374] In a further embodiment, the composition further comprises sucrose. In a particular embodiment, sucrose is present at a concentration of about 70 mg / mL. In a particular embodiment, sucrose is present at a concentration of 7% (w / v).

[0375] In a further embodiment of the present invention, the composition further comprises polysorbate 80. In certain embodiments, polysorbate 80 is present at a concentration of about 0.2 mg / mL. In certain embodiments, polysorbate 80 is present at a concentration of 0.02% (w / v).

[0376] In certain embodiments, the composition comprises 10 mM L-methionine, 10 mM histidine, pH 5.5, 7% sucrose, 0.02% polysorbate 80, and 130 mg / mL anti-PD-1 antibody or an antigen-binding fragment thereof.

[0377] In certain embodiments, the composition comprises 10 mM L-methionine, 10 mM histidine, pH 5.5, 7% sucrose, 0.02% polysorbate 80, and 165 mg / mL anti-PD-1 antibody or an antigen-binding fragment thereof.

[0378] In a further embodiment, the composition further comprises 2000 U / ml PH20 variant 2. In a further embodiment, the composition further comprises 2000 U / ml rHuPH20.

[0379] In one embodiment, the composition is contained in a vial. In another embodiment, the composition is contained in one or more prefilled syringes. In one embodiment, the composition is contained in two prefilled syringes.

[0380] In any kit or composition of the present invention, the anti-PD-1 antibody or antigen-binding fragment can be any of the antibodies or antigen-binding fragments described in Section II, "PD-1 Antibodies and Antigen-Binding Fragments Useful in the Present Invention" of the specific embodiments of the present invention. In one embodiment, the anti-PD-1 antibody or an antigen-binding fragment thereof is pembrolizumab. In another embodiment, the anti-PD-1 antibody or an antigen-binding fragment thereof is a pembrolizumab variant.

[0381] In any kit or composition of the present invention, the anti-PD-1 antibody or an antigen-binding fragment thereof can be co-formulated with the human hyaluronidase described in Section III.

[0382] These and other aspects of the present invention, including the exemplary specific embodiments listed below, will be apparent from the teachings contained herein.

[0383] General methods

[0384] Standard methods in molecular biology are described in Sambrook, Fritsch and Maniatis (2nd edition 1982 & 1989, 3rd edition 2001), Molecular Cloning, A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY; Sambrook and Russell (2001) Molecular Cloning, 3rd edition, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY; Wu (1993) Recombinant DNA, Vol. 217, Academic Press, San Diego, CA). Standard methods are also presented in Ausbel et al., (2001) Current Protocols in Molecular Biology, Vols. 1-4, John Wiley and Sons, Inc. New York, NY, which describe cloning and DNA mutagenesis in bacterial cells (Vol. 1), cloning in mammalian cells and yeast (Vol. 2), glycoconjugates and protein expression (Vol. 3) and bioinformatics (Vol. 4).

[0385] All publications mentioned herein are incorporated herein by reference to describe and disclose methods and materials that may be related to the present invention.

[0386] After describing various embodiments of the present invention with reference to the accompanying drawings, it should be understood that the present invention is not limited to these exact embodiments, and various changes and modifications can be made by those skilled in the art without departing from the scope or spirit of the present invention as defined in the appended claims.

[0387] Example 1

[0388] Population PK model development

[0389] Currently, pembrolizumab is approved for use in multiple cancer indications at doses of 200 mg or 2 mg / kg Q3W or 400 mg Q6W administered by IV infusion. An alternative subcutaneous ("SC") formulation using hyaluronidase would provide convenience and flexibility for patients and prescribers. A Phase I clinical study was conducted to evaluate the bioavailability of an SC formulation of pembrolizumab and the hyaluronidase PH20 variant 2 combination ("SC pembrolizumab-HLN" or "pembrolizumab-HLN SC") at two different solution strengths of pembrolizumab (concentration 165 mg / ml (arm 1) and 130 mg / ml (arm 2)). In this study, subcutaneous doses (650 mg over a 6-week dosing interval (cycle), providing pembrolizumab at two different concentrations / volumes (as shown in Table 6)) were used and compared to the IV dose of pembrolizumab (400 mg over a 6-week dosing interval) to evaluate the bioavailability of two different subcutaneous solution strengths of pembrolizumab (in two independent study arms).

[0390]

[0391] Both SC and IV pembrolizumab were administered to patients with advanced melanoma or non-small cell lung cancer (NSCLC) or renal cell carcinoma (RCC) in each study arm in the crossover design shown below: Cycle 1: SC 650 mg; Cycle 2: IV 400 mg; Cycle 3: SC 650 mg; Cycles 4 - 18: IV 400 mg.

[0392] Eligible patients were aged ≥ 18 years, had unresectable stage III or IV melanoma unsuitable for local therapy, measurable disease according to RECIST v1.1, Eastern Cooperative Oncology Group performance status score 0 or 1, or had not received prior therapy for advanced disease (except BRAF / MEK inhibitors for BRAF V600 mutated disease and ≥ 4 weeks of prior adjuvant or neoadjuvant therapy received from randomization); had a histologically or cytologically confirmed diagnosis of metastatic (stage IV [M1a, M1b, or M1c according to current AJCC criteria]) non-squamous NSCLC without EGFR, ALK, or ROS1 genomic tumor abnormalities, or metastatic (stage IV [M1a, M1b, or M1c according to current AJCC criteria]) squamous NSCLC; had a histologically or cytologically confirmed diagnosis of RCC with a clear cell component, with or without sarcomatoid features, unresectable, locally advanced / metastatic (i.e., stage IV RCC according to AJCC); and had not received prior systemic therapy for advanced ccRCC.

[0393] For participants with NSCLC or RCC, treatment with pembrolizumab-HLN SC and pembrolizumab IV consisted of combinations with standard therapies as shown below:

[0394] NSCLC: Pemetrexed and platinum (carboplatin or cisplatin) chemotherapy for 1L treatment of patients with metastatic non-squamous NSCLC without EGFR or ALK genomic tumor aberrations; or carboplatin and paclitaxel or nab-paclitaxel for 1L treatment of patients with metastatic squamous NSCLC.

[0395] RCC: Axitinib for 1L treatment of patients with advanced RCC.

[0396] Table 7: Study Interventions

[0397]

[0398]

[0399] Serum concentration data from 81 subjects collected during Cycle 1 (i.e., Weeks 0-6) and Cycle 2 (i.e., Weeks 12) of the Phase I clinical trial were used to characterize the PK of the SC pembrolizumab co-formulation with hyaluronidase, as well as PK data for widely historic pembrolizumab IV using population PK analysis. A non-linear mixed effects model was used for the Phase 1 data, which were from prior data of a previously established pembrolizumab reference PK model. The reference pembrolizumab PK model was based on pembrolizumab PK data collected from 2993 patients with various cancers who received doses of pembrolizumab at 1 to 10 mg / kg Q2W, 2 to 10 mg / kg Q3W, or 200 mg Q3W in Phase I or Phase III clinical studies. PK parameters for the absorption phase of SC-administered pembrolizumab were estimated from the Phase I data, and any differences between the two solution strengths were also evaluated. Distribution and elimination parameters (clearance (CL), central volume of distribution (Vc), inter-compartmental clearance (Q), and peripheral volume of distribution (Vp)) were determined by the reference IV model, as these are the same for both IV and SC administration. Given the small sample size and short duration of SC administration in the study, i.e., two treatment cycles, parameters describing the time-dependence and impact of patient baseline characteristics on pembrolizumab PK were also determined from the previously established reference IV PK model.

[0400] A new population PK model was developed that can simultaneously describe pembrolizumab PK after IV or SC (with hyaluronidase). The final parameter estimates for the combined SC and IV population PK models are shown in Table 8. The absorption phase for SC administration was characterized by first-order absorption rate (ka) and bioavailability (F) parameters. The distribution and elimination phases were described by a two-compartment model with a time-dependent clearance and a fixed effect of body weight, as historically established in the reference pembrolizumab PK model. The inclusion of covariates effects for SC solution strength was not statistically significant, indicating no meaningful differences in bioavailability and absorption rate between the two pembrolizumab-HLN SC solution strengths (i.e., Arm 1 165 mg / mL and Arm 2 130 mg / ml). Goodness-of-fit evaluations indicated no structural bias as a function of drug concentration or time. The analysis showed that the predicted bioavailability of pembrolizumab-HLN was 57% (range: 38% to 75%) when administered SC. The median time to reach maximum pembrolizumab serum concentration in the subcutaneous formulation with hyaluronidase was estimated to be 4 days (range, 2 to 35 days). Additionally, in the Phase I study, anti-drug antibodies (ADA) were observed in 1 / 83 (<2%) subjects.

[0401]

[0402]

[0403] Results from the ongoing Phase I study indicate no new systemic safety signals for SC pembrolizumab-HLN. All injection site reactions were not severe, mostly mild (Grade 1) and were effectively managed. The overall safety profile during SC pembrolizumab-HLN and pembrolizumab IV cycles was consistent with the known safety profiles of pembrolizumab monotherapy and standard therapies administered in participants with NSCLC and RCC.

[0404] The two tested SC solution strengths of pembrolizumab-HLN (130 mg / ml and 165 mg / ml) had similar absorption PK. SC administration of pembrolizumab-HLN was well tolerated and had no significant ADA. Based on an analysis of PK model simulations using the estimated bioavailability and inter-subject variability from this study, it is believed that a subcutaneous dose of 760 to 790 mg Q6W of pembrolizumab-HLN should result in similar exposure to the approved dose of 400 mg Q6W pembrolizumab IV.

[0405] Example 2

[0406] SC pembrolizumab co-formulated with hyaluronidase across multiple tumor types was evaluated using a once-weekly (Q6W) dosing schedule based on modeling and simulation with 400 mg Q6W IV as the reference regimen.

[0407] Pembrolizumab (an anti-PD-1 checkpoint inhibitor) currently approved for multiple cancer indications has been shown to be safe and effective when administered by intravenous infusion (IV) at doses of 200 mg Q3W, 2 mg / kg Q3W, or 400 mg Q6W. The robust characteristics of pembrolizumab pharmacokinetics (PK) and exposure (concentration)-response (E-R) relationships in terms of efficacy and safety allow the use of model-based approaches to support alternative administration routes of pembrolizumab. Previously, KN-252 efficacy data in 294 patients with advanced melanoma showed a flat exposure-response relationship in the C trough range of 3.5 - 32.5 μg / ml at the 200 mg Q3W IV pembrolizumab dose. KN-555 data in 89 patients with advanced melanoma showed that the observed C trough at 400 mg Q6W IV was exactly within the previously established flat exposure-response relationship. Therefore, the C trough at 400 mg Q6W IV was sufficient to maximize the efficacy of pembrolizumab Figure 1 .

[0408] KEYNOTE-555 (NCT03665597) is an open-label, multi-cohort, phase 1 clinical trial investigating pembrolizumab in patients with advanced melanoma. Cohort B studied the pharmacokinetics, efficacy, and safety of intravenous administration of pembrolizumab at a dose of 400 mg every 6 weeks. 101 patients were screened and treated. Clinically meaningful objective response rates and duration of continuous progression-free survival were observed within the expected range for front-line pembrolizumab. The objective response rate was 50.5% (95% CI, 40.4 - 60.6); 19 patients (18.8%) had a complete response and 32 (31.7%) had a partial response. The median duration of response was not reached (NR; range, 2.4+ to 21.0+ months). The median progression-free survival was 13.8 months (95% CI, 4.1 - NR). The observed pharmacokinetic exposure was consistent with model predictions for pembrolizumab 400 mg Q6W IV and within the exposure of other approved and tested regimens (2 mg / kg or 200 mg Q3W IV and 10 mg / kg Q2W IV) (see Figure 2). Safety in Cohort B was comparable to the known properties of the pembrolizumab 200 mg Q3W IV regimen in melanoma. Grade 3 - 5 treatment-related adverse events occurred in 13 patients (12.9%). Three patients (3.0%) discontinued treatment due to treatment-related adverse events.

[0409] The phase 2 KEYNOTE-B68 trial investigated the efficacy and safety of intravenous (IV) pembrolizumab 400 mg every 6 weeks in patients (pts) with relapsed / refractory (R / R) classical Hodgkin lymphoma (cHL) or primary mediastinal B-cell lymphoma (PMBCL). At data cutoff, 66 pts were enrolled (60 R / R cHL, 6 R / R PMBCL). Median follow-up (range) was 8.9 months (1-15.9) for pts with R / R cHL and 10.6 months (5.1-15.4) for pts with R / R PMBCL. The objective response rate (ORR) was 65% (95% confidence interval [CI], 51.6–76.9 [33.3% complete response {CR}; 31.7% partial response {PR}]) for pts with R / R cHL and 50% (95% CI, 11.8–88.2 [33.3% CR; 16.7% PR]) for pts with R / R PMBCL. Drug-related adverse events (AEs) occurred in 24 pts (40%) with R / R cHL and 2 pts (33.3%) with R / R PMBCL. Grade ≥3 drug-related AEs occurred in 3 pts (5%) with R / R cHL and 1 pt (16.7%) with R / R PMBCL. Immune-mediated AEs occurred in 13 pts (21.7%) with R / R cHL and 1 pt (16.7%) with R / R PMBCL. Grade ≥3 immune-mediated AEs occurred in 2 pts (3.3%) with R / R cHL. At approximately 9 months of follow-up, pembrolizumab 400 mg Q6W IV demonstrated robust antitumor activity in pts with R / R cHL and R / R PMBCL and no new safety concerns. The ORR observed in R / R cHL and R / R PMBCL was similar to that of pembrolizumab 200 mg Q3W IV.

[0410] PK model-based simulations were conducted to select the subcutaneous (SC) dose, aiming for consistency of SC PK exposure profiles with those of the approved 400 mg Q6W IV dose and overall exposure profiles based on the clinical experience with pembrolizumab IV. The simulations were performed on a pooled dataset of a phase 1 study evaluating pembrolizumab-HLN SC (total N = 3105 subjects) and a reference pembrolizumab IV PK dataset of 2993 subjects with melanoma or non-small cell lung cancer (NSCLC) from previous phase I and III trials of IV pembrolizumab.

[0411] The serum concentrations of pembrolizumab-HLN SC in the dose range of 600 mg to 1200 mg Q6W and the dose of 400 mg Q6W of pembrolizumab IV from Cycle 1 to Cycle 3 (18 weeks, reaching steady state) were simulated using the SC and IV PK combination model (as described in Table 8), including the estimated population mean PK parameters and the between-subject variability and residuals for each parameter. For each subject in the dataset, the simulated trough concentration (C trough ) and the area under the curve (AUC) exposure at the end of the dosing interval were determined at Cycle 1 (first dose) and Cycle 3 (representing steady state). C trough and AUC 0-6周 represent PK exposure metrics and are considered drivers of pembrolizumab efficacy. Cycle 1 represents the PK exposure achieved after administration of the first dose. Cycle 3 represents the PK exposure achieved at steady state, and then these exposures will be maintained during treatment. The geometric mean (GM) of C trough and AUC 0-6周 was calculated for each SC dose of pembrolizumab and the 400 mg IV dose. Then, the geometric mean ratio (GMR) (as the ratio of the GM of each formulation group) of SC versus IV pembrolizumab for both of these PK exposure metrics was calculated for treatment Cycles 1 and 3.

[0412] Simulations based on the PK model showed that the dose range of 600 to 1000 mg Q6W of pembrolizumab-HLN SC (using hyaluronidase PH20 variant 2) resulted in exposure comparable to the approved dose of 400 mg Q6W of pembrolizumab administered by IV infusion, and is safe given the highest clinically evaluated dose of 10 mg / kg Q2W IV with established safety. In principle, given that the exposure-response relationship for the efficacy and safety of pembrolizumab has been well established, similar PK exposure would result in similar efficacy and safety of pembrolizumab. Therefore, SC doses in this range are expected to have comparable efficacy and safety to 400 mg Q6W IV.

[0413] For the lower limit of exposure (AUC or C trough ), it is appropriate to use the lower limit of the 90% CI of the generally accepted GMR > 0.8 or so to determine non-inferiority. This will determine that the efficacy of a certain dose of pembrolizumab administered subcutaneously co-formulated with hyaluronidase is not inferior to that of the dose of pembrolizumab (400 mg Q6W) administered by the IV route (“IV pembrolizumab”).

[0414] The developed simulations showed that SC pembrolizumab-HLN doses of 600 mg Q6W or higher all had an average SC:IV C troughRatio, and SC pembrolizumab-HLN doses of 600 mg Q6W or higher all had an average SC:IV AUC ratio greater than 0.8. See Tables 9A and 9B. Tables 9A-B show the C at Cycle 1 of different evaluated Q6W SC doses relative to 400 mg Q6W IV pembrolizumab trough and the C at steady state trough (C trough ss) SC (subcutaneous):IV (intravenous) geometric mean (GM) ratio (GMR). Tables 9C-D show the SC:IV GMR of AUC at Cycle 1 (AUC.C1) and at steady state (AUC.ss). 0-6周

[0415] Table 9A: Passed GM and GMR of C trough

[0416]

[0417]

[0418] Table 9B: Passed GM and GMR of C troughss

[0419]

[0420] Table 9C: Passed GM and GMR of AUC at Cycle 1

[0421]

[0422] Table 9D: Passed GM and GMR of AUCss

[0423]

[0424]

[0425] The dose range of 760 to 790 mg Q6W of SC pembrolizumab-HLN had a PK exposure similar to that of the approved 400 mg Q6W pembrolizumab dose by IV infusion. Based on the analysis conducted, it is expected that the efficacy of the 760 to 790 mg Q6W SC pembrolizumab-HLN dose will be similar to that of 400 mg Q6W IV pembrolizumab, as follows:

[0426] Throughout the entire treatment duration, it is expected that the C of the 760 to 790 mg Q6W SC dose will be approximately 25 to 30% higher than that of 400 mg Q6W IV. Additionally, at Cycle 1 and steady state, the distribution of C trough largely overlaps between SC and IV. See trough ​​​​Figure 3A and Figure 3B Over the entire treatment duration, the AUC of the 760 to 790 mg Q6W SC dose is expected to be approximately 2 to 10% higher than the AUC 0-6周 exposure of the 400 mg Q6W IV. In addition, at Cycle 1 and at steady state, the distribution of the AUC 0-6周 mostly overlaps between SC and IV. See 0-6周 . Figure 4A and Figure 4B .

[0427] In the case of comparing SC pembrolizumab to IV pembrolizumab, it is important to note the key differences inherent in the PK properties between SC and IV administration. Generally, for comparable doses (adjusted for bioavailability), the concentration following SC administration gradually increases over ~4 days and the peak concentration (C max ) is much lower than the C max reached at the end of the IV infusion. Specifically, the C max at the 760 to 790 mg Q6W SC dose is expected to be substantially lower (decreased by ~38 to 35% at Cycle 1 and by ~25 to 22% at steady state) compared to the C max reached with 400 mg Q6W IV. See Figure 5 . Thus, within the pembrolizumab-HLN SC dose range, the C max over the entire course of treatment is not expected to increase compared to the approved IV dose of 400 mg Q6W. In addition, all PK exposures of 760 to 790 mg Q3W SC pembrolizumab-HLN will surely be far lower than 5 times the high dose / exposure of 10 mg / kg Q2W IV, which is the highest clinically evaluated dose with established safety. Therefore, the safety profile following SC administration of 760 to 790 mg Q6W SC pembrolizumab-HLN is not likely to be different from the safety profile previously established for the identified 400 mg Q6W IV pembrolizumab dose, and thus no further quantitative assessment of the exposure upper limit is evaluated.

[0428] The selected dose for the Phase III clinical trial using the hyaluronidase PH20 variant 2 and pembrolizumab SC is 790 mg Q6W. Figure 3A and Figure 3B summarize the results of population simulations, which include the variability of the C trough for the 790 mg Q6W SC and 400 mg Q6W IV doses of pembrolizumab. The simulations developed suggest that the 790 mg Q6W SC dose results in a range of C trough in different patients, which generally overlaps with the 400 mg Q6W IV dose. Figure 4A andFigure 4B Summarizes the results of population simulations that included the AUC variability for pembrolizumab 790 mg Q6W SC and 400 mg Q6W IV doses. Simulations showed that the 790 mg Q6W SC dose produced a range of AUCs in different patients that generally overlapped with the 400 mg Q6W IV dose. 0-6周 Simulations showed that the 790 mg Q6W SC dose produced a range of AUCs in different patients that generally overlapped with the 400 mg Q6W IV dose. 0-6周 that generally overlapped with the 400 mg Q6W IV dose. Figure 5 Summarizes the results of population simulations that included the C variability for pembrolizumab 790 mg Q6W SC and 400 mg Q6W IV doses. Simulations showed that the 790 mg Q6W SC dose produced a range of Cs in different patients that were generally lower than the 400 mg Q6W IV dose. max Simulations showed that the 790 mg Q6W SC dose produced a range of Cs in different patients that were generally lower than the 400 mg Q6W IV dose. max that were generally lower than the 400 mg Q6W IV dose. Figure 3A and Figure 3B , Figure 4A and Figure 4B describe the distributions (5th, 25th, 50th, 75th, and 95th percentiles) of C trough and AUC 0-6周 at cycle 1 and steady state, respectively, and Figure 5 describes the distribution (5th, 25th, 50th, 75th, and 95th percentiles) of C max at steady state for pembrolizumab doses of 790 mg Q6W SC and 400 mg Q6W IV using PK model-based simulations. Simulated PK exposure metrics in 3105 subjects are shown.

[0429] In summary, model-based simulations supported by Tables 9A-D, Figure 3A , 3B , 4A, 4B, and 5 indicate that the 790 mg Q6W dose of pembrolizumab-HLN administered SC should produce optimal PK exposure properties similar to those of the approved 400 mg Q6W dose of pembrolizumab IV, maintaining efficacy while remaining within the clinical safety margins.

[0430] Example 3

[0431] A Phase 3, randomized, open-label clinical study evaluating the pharmacokinetics and safety of subcutaneous pembrolizumab co-formulated with hyaluronidase (pembrolizumab-HLN) compared to intravenous pembrolizumab in the first-line treatment of participants with metastatic non-small cell lung cancer in combination with chemotherapy

[0432] This is a Phase 3, randomized, active-control, parallel-group, multi-center, open-label study of pembrolizumab-HLN and platinum doublet chemotherapy compared to pembrolizumab IV and platinum doublet chemotherapy in participants with untreated metastatic NSCLC.

[0433] After a screening period of up to 28 days, participants will be randomly assigned to Arm 1 or Arm 2 at a ratio of 2:1. One cycle is 6 weeks. Arm 1 is the combination of pembrolizumab-HLN Q6W and platinum doublet chemotherapy for up to 18 cycles. Arm 2 is the combination of pembrolizumab IV Q6W and platinum doublet chemotherapy for up to 18 cycles.

[0434] The platinum doublet chemotherapy is as follows: For non-squamous NSCLC: Up to 4 infusions of pemetrexed Q3W with platinum chemotherapy (cisplatin Q3W or carboplatin Q3W), followed by pemetrexed maintenance until one of the conditions for stopping the study intervention is met. For squamous NSCLC: Up to 4 infusions of carboplatin Q3W with taxane (paclitaxel Q3W or nab-paclitaxel [days 1, 8, 15, 22, 29, and 36 of the first and second cycles]).

[0435] Randomization will be stratified by ECOG performance status, histology, PD-L1 TPS, and region. Each participant will receive the study intervention until one of the conditions for stopping the study intervention is met.

[0436] The dual primary objective of this study is to compare the AUC of pembrolizumab-HLN versus pembrolizumab IV in cycle 1 under the Q6W dosing regimen 0-6周 and the C trough at steady state (cycle 3). Non-inferiority will be evaluated using a non-inferiority margin of 0.8. Tumor response will be evaluated according to the RECIST 1.1 adaptation. Safety assessments will also be conducted.

[0437] The combination of pembrolizumab IV and platinum doublet chemotherapy will be used as first-line treatment for patients with metastatic NSCLC. This study will primarily compare the pembrolizumab exposure between pembrolizumab-HLN (Arm 1) and pembrolizumab IV (Arm 2), both in combination with chemotherapy, in treatment-naive participants with metastatic NSCLC under the Q6W dosing regimen. Other descriptive comparisons of pembrolizumab C trough will also be made between pembrolizumab-HLN and pembrolizumab 200 mg IV Q3W using model-based exposure.

[0438] Study objectives and endpoints:

[0439] In treatment-naive participants with metastatic NSCLC:

[0440] Table 10

[0441]

[0442]

[0443] Table 11: Overall study design

[0444]

[0445] AUC = Area Under the Curve; IV = Intravenous; Q3W = Every 3 weeks; Q6W = Every 6 weeks; SC = Subcutaneous. a Cycle = 6 weeks b For non-squamous, AUC 5 mg / ml / min; for squamous, AUC 6 mg / ml / min

[0446] Rationale for Pharmacokinetic Endpoints

[0447] The first cycle AUC 0-6周 and steady state (cycle 3) C trough of the dual primary endpoints were used to compare pembrolizumab exposure between pembrolizumab-HLN and pembrolizumab IV Q6W. The first cycle AUC 0-6周 is the most conservative PK value to ensure non-inferiority of SC exposure relative to IV. Due to accumulation, any exposure differences between SC and IV administration of pembrolizumab should decrease after multiple doses. Therefore, demonstrating non-inferiority of SC AUC exposure in the first cycle also implies non-inferiority at steady state. Based on our analysis, higher AUC exposure at steady state after SC dosing is not expected to exceed the established clinical safety boundary of pembrolizumab (i.e., the exposure of 10 mg / kg Q2W IV).

[0448] Steady state (cycle 3) C trough is the concentration at the end of the 6-week dosing interval after the third dose, which is the steady state for pembrolizumab. Since the pharmacological activity of mAb is mediated through direct interaction with specific targets, target saturation can be used as a surrogate for maximal pharmacological and therapeutic activity. At the approved IV dose, pembrolizumab exposure is expected to maintain PD-1 target saturation throughout the dosing interval, thereby maintaining efficacy. Therefore, the C trough of the approved IV dose of 400 mg Q6W can be considered a threshold above which target saturation and efficacy will be maintained. By demonstrating non-inferiority of pembrolizumab steady state (cycle 3) C trough for pembrolizumab-HLN, it can be inferred that it will maintain efficacy similar to that of pembrolizumab IV administration.

[0449] The secondary PK endpoints will be able to further characterize pembrolizumab exposure: for comparison with pembrolizumab IV Q6W: cycle 1: C max , C trough ; steady state (cycle 3): AUC 0-6周 and C max . For comparison with pembrolizumab IV Q3W: model-based C trough at cycle 1 and steady state.

[0450] PK model-based simulations showed that the 790 mg Q6W pembrolizumab dose resulted in exposures similar to those of the approved 400 mg Q6W pembrolizumab IV dose. The following are expected to retain efficacy for SC pembrolizumab at the 790 mg Q6W dose:

[0451] · The C trough is expected to be approximately 30% higher for the entire treatment duration at the 790 mg Q6W SC dose than the C trough of 400 mg Q6W IV. In addition, the C trough distribution overlapped between SC and IV at both cycle 1 and steady state (see Figure 3A and Figure 3B ).

[0452] · The AUC 0-6周 exposure is expected to be approximately 10% higher for the entire treatment duration at the 790 mg Q6W SC dose than the AUC 0-6周 exposure of 400 mg Q6W IV. In addition, the AUC 0-6周 distribution overlapped between SC and IV at both cycle 1 and steady state (see Figure 4A and Figure 4B ).

[0453] The following are expected to maintain safety for SC pembrolizumab at the 790 mg Q6W dose:

[0454] · The C max is expected to be lower than the C max achieved at 400 mg Q6W IV (approximately 35% lower at cycle 1 and approximately 22% lower at steady state) ( Figure 5 ). Thus, there is no expected increase in C max relative to the approved 400 mg Q3W IV dose over the entire treatment duration.

[0455] · All SC exposures (C max , C avg , C trough ) are expected to remain below the Cmax and initial concentration of 400 mg Q6W IV and well below the highest doses and exposures established for clinical safety (i.e., 10 mg / kg Q2W) over the 6-week dosing interval and the duration of the entire treatment.

[0456] In summary, as discussed above, the developed model-based simulations showed that the 790 mg Q6W dose of subcutaneous pembrolizumab-HLN should result in optimal PK exposure profiles similar to those of the approved 400 mg Q6W dose of pembrolizumab, thus maintaining efficacy while remaining within the clinical safety margins.

[0457] Example 4

[0458] Based on the evaluation using modeling and simulation with Q6W SC pembrolizumab-HLN as the reference regimen, a dosing schedule of SC pembrolizumab-HLN every three weeks (Q3W) in multiple tumor types was developed.

[0459] The developed PK model-based simulation showed that the dosing regimen of 790 mg Q6W of SC pembrolizumab with hyaluronidase PH20 variant 2 resulted in similar exposure to the 400 mg Q6W dose of approved pembrolizumab administered by IV infusion. Given that the exposure-response relationship for the efficacy and safety of pembrolizumab has been well established, in principle, similar PK exposure leads to similar efficacy and safety of pembrolizumab. Using such exposure-matching methods, the Q3W dose of SC pembrolizumab-HLN was selected using PK model-based simulation, with the Q6W SC pembrolizumab-HLN dose as the reference.

[0460] PK model-based simulation was performed to select the Q3W SC dose, aiming for consistency in the PK exposure profile of the SC dose with that of the Q6W SC dose and the overall exposure profile based on the clinical experience with pembrolizumab IV. This simulation was performed on the pooled dataset of the phase 1 study of pembrolizumab-HLN SC (a total of N = 3105 subjects), and the reference pembrolizumab IV PK dataset included 2993 melanoma or non-small cell lung cancer (NSCLC) subjects from previous phase I and III trials of IV pembrolizumab.

[0461] Using the combined SC and IV PK model (described in Table 8), the serum concentration of pembrolizumab was simulated for a dose range of 380 mg to 410 mg Q3W of pembrolizumab-HLN SC from cycle 1 to cycle 6 or cycle 3 (18 weeks, reaching steady state) and the corresponding Q6W SC dose range that is twice the Q3W dose. The model contains estimates of population-average PK parameters and between-subject variability and residuals for each parameter. For each subject in the dataset, the simulated trough concentration (C trough ) and the area under the curve (AUC) exposure were determined at both the end of the dosing interval at cycle 1 (first dose) and at steady state (cycle 6 for Q3W or cycle 3 for Q6W). C trough and AUC indicate PK exposure metrics and are considered drivers of pembrolizumab efficacy. Cycle 1 represents the PK exposure achieved after administration of the first dose. The PK exposure reached at steady state is then the exposure that will be maintained throughout the treatment period. The C troughThe geometric mean (GM) of and AUC. Then, for the 1st treatment cycle and steady state, calculate the geometric mean ratio (GMR) of SC vs IV pembrolizumab for both of these PK exposure metrics (as the GM ratio for each formulation group).

[0462] The simulations developed showed that the Q3W SC pembrolizumab-HLN dose corresponding to half the Q6W SC pembrolizumab-HLN dose had over the Q6W SC dose from cycle 1 to steady state for C trough (mean SC-Q3W:SC-Q6W C trough ratio greater than 1), comparable AUC exposure at cycle 1 (mean SC-Q3W:SC-Q6W AUC ratio greater than 0.8) and the same AUC exposure at steady state (mean SC-Q3W:SC-Q6W AUC ratio of 1). See Tables 12A-D. Tables 12A-B show C trough and C trough at steady state (C troughss ) Q3W:Q6W geometric mean ratio (GMR). In Table 12A, the sampling period for the SC-Q3W regimen was 3 weeks and the sampling period for the SC-Q6W reference was 6 weeks. In Table 12B, the sampling period for the SC-Q3W regimen was 6 weeks and the sampling period for the SC-Q6W reference was 6 weeks. Tables 12C-D show the AUC 0-6周 Q3W:Q6W GMR at cycle 1 (AUC.C1), cycle 2 and steady state (AUC.ss) at different evaluated Q3W and Q6W SC doses of pembrolizumab-HLN. In Tables 12C-D, the sampling period for the SC-Q3W regimen was 6 weeks and the sampling period for the SC-Q6W reference was 6 weeks.

[0463] Table 12A: Passed trial GM and GMR of C trough at 3W in cycle 1

[0464]

[0465] Table 12B: Passed trial GM and GMR of C trough ss

[0466]

[0467] Table 12C: Passed trial GM and GMR of AUC.6w ss

[0468]

[0469] Table 12D: GM and GMR Passing the Test in the 1st / 2nd Cycle of AUC.6w

[0470]

[0471] The selected dose of pembrolizumab SC using hyaluronidase PH20 variant 2 is 790 mg Q6W. The corresponding selected dose of pembrolizumab SC using hyaluronidase PH20 variant 2 at Q3W is 395 mg. Based on our analysis, it is expected that SC pembrolizumab-HLN retains efficacy at a dose of 395 mg Q3W:

[0472] It is expected that C at a SC dose of 395 mg Q3W throughout the treatment duration trough is about 15 to 60% higher than that of 790 mg Q6W SC. In addition, C trough distributions overlap mostly between SC-Q3W and SC-Q6W both in the 1st cycle and at steady state. See Figure 6A and Figure 6B .

[0473] It is expected that the AUC at a SC dose of 395 mg Q3W throughout the treatment duration 0-6周 exposure is similar to that of 790 mg Q6W SC. In addition, the AUC 0-6周 distributions overlap mostly between SC-Q3W and SC-Q6W both in the 1st cycle and at steady state. See Figure 7A and Figure 7B .

[0474] It is expected that SC pembrolizumab-HLN using a dose of 395 mg Q3W will maintain safety, considering that C max will remain below the 790 mg Q6W dose throughout the treatment period from the 1st cycle to steady state.

[0475] Figure 6A - 6B summarizes the results of population simulations, which include the variability of C trough for the 395 mg Q3W SC and 790 mg Q6W SC doses of pembrolizumab-HLN. The developed simulations show that the 395 mg Q3W SC dose results in a C trough range for different patients that generally overlaps with the 790 mg Q6W SC dose. Figure 7A and Figure 7B summarize the results of population simulations, which include the variability of AUC 0-6周 for the 395 mg Q3W SC and 790 mg Q6W SC doses of pembrolizumab-HLN. The simulations show that the 395 mg Q3W SC dose results in an AUC 0-6周The range typically overlaps with the 790 mg Q6W SC dose. Figure 6A and Figure 6B , Figure 7A and Figure 7B describe the distribution of C trough and AUC 0-6周 at Cycle 1 and steady state (percentiles 5, 25, 50, 75, and 95), using PK model-based simulations at the 395 mg Q3W and 790 mg Q6W SC doses of pembrolizumab-HLN. Simulated PK exposure metrics in 3105 subjects are shown.

[0476] In summary, as supported by Tables 12A-D, Figure 6A , 6B , 7A, and 7B, model-based simulations suggest that the 395 mg Q3W dose of pembrolizumab-HLN administered SC should result in optimal PK exposure properties, which are consistent with pembrolizumab-HLN SC 790 mg Q6W, thereby maintaining efficacy while remaining within the clinical safety margins. Additionally, based on our analysis, 300 - 500 mg Q3W SC pembrolizumab-HLN doses are also expected to be safe and effective and have PK exposure comparable to 600 - 1000 mg Q6W SC pembrolizumab-HLN doses.

[0477] Approximately 50 participants with unresectable advanced melanoma were assigned to Arm 4 of the Phase I clinical trial in Table 7 (165 mg / mL pembrolizumab-HLN SC, 2.39 ml). The treatment cycle was 21 days (Q3W), and the dose was 395 mg of pembrolizumab, using pembrolizumab-HLN SC. PK exposure metrics, including C trough , C max and AUC, will be monitored at Cycle 1 and steady state (Cycle 6). Overall survival, safety, and anti-pembrolizumab antibodies will be monitored in Arm 4. The trough concentration of pembrolizumab (C trough ) after pembrolizumab-HLN SC treatment will be measured by collecting patient PK samples at any time on Day 1 before dosing (0 - 3 hours) in Cycle 1 and Cycle 6; on Days 2, 4, 6, 10, and 15 in Cycle 1 and Cycle 6. The maximum plasma concentration of pembrolizumab (C max)。The maximum area under the curve (AUC) of pembrolizumab after pembrolizumab-HLN SC treatment will be measured by collecting patient PK samples at pre-dose (0 - 3 hours) on Day 1 of Cycle 1 and Cycle 6; and at any time on Days 2, 4, 6, 10, and 15 of Cycle 1 and Cycle 6.

[0478] Example 5

[0479] Based on an evaluation using PK modeling and simulation with 400 mg Q6W IV as the reference regimen, a dosing schedule of SC pembrolizumab (without hyaluronidase) every six weeks (Q6W) in multiple tumor types was developed.

[0480] PK data from the Phase 1 study of SC pembrolizumab-HLN as described above were generally consistent with PK data from the Phase 1 study of SC pembrolizumab without hyaluronidase (the same formulation in KEYNOTE-555 Cohort A, Table 6 without hyaluronidase). The results of comparing these two studies are shown in Table 13. Thus, the bioavailability ranges of these formulations largely overlap, and the data confirm that the addition of hyaluronidase does not significantly affect the PK of SC pembrolizumab administration. Therefore, the PK exposures (C trough , AUC, C max ) profiles are expected to be similar. Thus, by extrapolation, it is also expected that the selected Q6W dose of 790 mg and the dose range of 600 to 1000 mg identified for SC pembrolizumab with hyaluronidase will be safe and effective when applied to SC pembrolizumab without hyaluronidase.

[0481] Table 13: Pharmacokinetic data of subcutaneous formulations of pembrolizumab-HLN and pembrolizumab

[0482]

[0483] Example 6

[0484] Pharmacokinetics of pembrolizumab between different tumor types and combination therapies

[0485] During the clinical development of pembrolizumab, extensive historical and current data from multiple indications and treatment settings have shown that both PK and immunogenicity are generally consistent across different tumor types and between monotherapy and combination therapies. In addition, based on 8 randomized dose comparisons in melanoma and non-small cell lung cancer (NSCLC), the exposure-response relationships for both efficacy and safety of pembrolizumab have been well established and have been shown to be flat over a >5-fold dose / exposure range in clinical studies ranging from 2 mg / kg Q3W to 10 mg / kg Q2W IV. Based on pooled data analysis of treatment arms from various studies, consistent, flat exposure-response relationships have also been observed for other indications (e.g., head and neck squamous cell carcinoma (HNSCC), classical Hodgkin lymphoma (cHL), UC (urothelial carcinoma), GC (gastric cancer), PMBCL (primary mediastinal large B-cell lymphoma), and MSI-H cancers) Figure 8 )

[0486] PK Consistency Across Tumor Types

[0487] As the basis for the pembrolizumab label, the reference PK model for pembrolizumab is based on a definitive analysis characterizing the PK profile of pembrolizumab from a robust dataset of 2,993 melanoma or NSCLC participants from KEYNOTE-001, KEYNOTE-002, KEYNOTE-006, KEYNOTE-101, and KEYNOTE-024 (including doses of 2 mg / kg Q3W, 10 mg / kg Q3W, 10 mg / kg Q2W, and 200 mg Q3W). In the reference analysis, tumor type had no meaningful effect on PK. The model also evaluated the PK parameter consistency between other approved indications (HNSCC, UC, GC, MSI-H cancers, cHL, PMBCL, HCC, and cervical cancer). Data from participants with these cancer types were added to the reference dataset (based on melanoma and NSCLC), and the parameters of the reference model were re-estimated. This updated PK analysis including several approved tumor types yielded model parameter estimates consistent with the reference analysis based on melanoma and NSCLC.

[0488] In addition, PK consistency between individual tumor types and the reference PK model (melanoma and NSCLC) has been evaluated by covering the concentrations observed within the 90% prediction intervals of the model. These analyses showed that the observed concentrations decreased within the predicted concentration ranges, regardless of weight-based or fixed dosing; the smallest differences were observed in hematologic malignancies (such as cHL and PMBCL) and were considered not clinically significant. This indicates that the PK model is able to adequately describe the PK of pembrolizumab across different tumor types, confirming the similarity of pembrolizumab PK across different indications following IV administration.

[0489] PK consistency between monotherapy and combination therapy

[0490] Chemotherapy is usually metabolized in the liver and generally has no effect on the disposition of monoclonal antibodies in the body. As described in the literature, monoclonal antibodies are mainly catabolized by the human reticuloendothelial system. Under normal circumstances, from a pharmacological perspective, the PK interaction between pembrolizumab and small molecules is not predictable because metabolic or transporter protein pathways are not involved in the disposition of pembrolizumab.

[0491] In multiple clinical studies where pembrolizumab was administered with chemotherapy (KEYNOTE-021, KEYNOTE-189: using carboplatin / cisplatin and pemetrexed; KEYNOTE-407: using carboplatin and paclitaxel / albumin-bound paclitaxel; KEYNOTE-048: using carboplatin / cisplatin and 5-FU; and KEYNOTE-426: using axitinib), no effect on the overall exposure of pembrolizumab was observed compared to the use of pembrolizumab monotherapy.

[0492] Applicability of pembrolizumab-HLN administration

[0493] In the context of SC administration, bioavailability and absorption are not expected to be affected by tumor type or combination therapy (Anselmo AC et al., Nat Rev Drug Discov. 2019;18:19-40), and given that the distribution and elimination phases are the same as for IV, PK consistency should be maintained between different tumor types and treatment settings. Therefore, based on studies of combination chemotherapy in NSCLC, the comparison of pembrolizumab PK parameters between SC and IV formulations is equally applicable to infer matching exposures between SC and IV, and thus to infer the bridging efficacy and safety of pembrolizumab indications.

Claims

1. A method of treating cancer in a human patient in need thereof, comprising subcutaneously administering to the patient a dose of about 600 mg to about 1000 mg of an anti-PD-1 antibody or an antigen-binding fragment thereof and human hyaluronidase every six weeks, wherein the anti-PD-1 antibody or an antigen-binding fragment thereof comprises: A light chain (LC) variable region comprising complementarity-determining regions (CDRs) LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1, LC-CDR2, and LC-CDR3 comprise the amino acid sequences shown in SEQ ID NOs: 1, 2, and 3, respectively, and a heavy chain (HC) variable region comprising CDR HC-CDR1, HC-CDR2, and HC-CDR3, wherein CDR HC-CDR1, HC-CDR2, and HC-CDR3 comprise the amino acid sequences shown in SEQ ID NOs: 6, 7, and 8, respectively.

2. A method of treating cancer in a human patient in need thereof, comprising subcutaneously administering to the patient a dose of about 600 mg to about 1000 mg of an anti-PD-1 antibody or an antigen-binding fragment thereof every six weeks, wherein the anti-PD-1 antibody or an antigen-binding fragment thereof comprises: A light chain (LC) variable region comprising complementarity-determining regions (CDRs) LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1, LC-CDR2, and LC-CDR3 comprise the amino acid sequences shown in SEQ ID NOs: 1, 2, and 3, respectively, and a heavy chain (HC) variable region comprising CDR HC-CDR1, HC-CDR2, and HC-CDR3, wherein CDR HC-CDR1, HC-CDR2, and HC-CDR3 comprise the amino acid sequences shown in SEQ ID NOs: 6, 7, and 8, respectively.

3. The method according to any one of claims 1-2, wherein the dose is 650 to 800 mg administered every six weeks.

4. The method according to any one of claims 1-2, wherein the dose is 700 to 800 mg administered every six weeks.

5. The method according to any one of claims 1-2, wherein the dose is 760 to 790 mg administered every six weeks.

6. The method according to any one of claims 1-2, wherein the dose is 790 mg administered every six weeks.

7. A method of treating cancer in a human patient in need thereof, comprising subcutaneously administering to the patient a dose of about 300 mg to about 500 mg of an anti-PD-1 antibody or an antigen-binding fragment thereof and human hyaluronidase every three weeks, wherein the anti-PD-1 antibody or antigen-binding fragment comprises: A light chain (LC) variable region comprising complementarity-determining regions (CDRs) LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1, LC-CDR2, and LC-CDR3 comprise the amino acid sequences shown in SEQ ID NOs: 1, 2, and 3, respectively, and a heavy chain (HC) variable region comprising CDR HC-CDR1, HC-CDR2, and HC-CDR3, wherein CDR HC-CDR1, HC-CDR2, and HC-CDR3 comprise the amino acid sequences shown in SEQ ID NOs: 6, 7, and 8, respectively.

8. The method according to claim 7, wherein the dose is 325 to 400 mg every three weeks.

9. The method according to claim 7, wherein the dose is 380 to 410 mg every three weeks.

10. The method according to claim 7, wherein the dose is 395 mg every three weeks.

11. The method according to any one of claims 1-10, wherein subcutaneous administration of the anti-PD-1 antibody or antigen-binding fragment thereof results in C of the antibody or antigen-binding fragment thereof trough within 20% of C of a 400 mg dose of the anti-PD-1 antibody or antigen-binding fragment thereof administered by intravenous (IV) administration every 6 weeks trough or at least the same as or up to 35% greater than it.

12. The method according to any one of claims 1-10, wherein subcutaneous administration of the anti-PD-1 antibody or antigen-binding fragment thereof results in an SC:IV C ratio of 0.8 to 1.6, 1.0 to 1.6, 1.1 to 1.6, 1.2 to 1.6, 1.3 to 1.6, 1.4 to 1.6, 1.2 to 1.5, 1.3 to 1.5, 1.4 to 1.5 or 1.3 to 1.4 compared to a 400 mg dose of the anti-PD-1 antibody or antigen-binding fragment thereof administered intravenously (IV) every 6 weeks. trough Ratio.

13. The method according to any one of claims 1-12, wherein subcutaneous administration of the anti-PD-1 antibody or antigen-binding fragment thereof results in an AUC of the anti-PD-1 antibody or antigen-binding fragment thereof at a dose of 400 mg administered by the intravenous (IV) administration route every 6 weeks (0-6周) that is at least 1.0-fold the AUC of the antibody or antigen-binding fragment thereof (0-6周) .

14. The method according to any one of claims 1-13, wherein the cancer is selected from the following: melanoma, non-small cell lung cancer, head and neck cancer, urothelial cancer, breast cancer, gastric cancer, gastroesophageal junction adenocarcinoma, multiple myeloma, hepatocellular carcinoma, Merkel cell carcinoma, renal cell carcinoma, endometrial cancer, cutaneous squamous cell carcinoma, non-Hodgkin lymphoma, Hodgkin lymphoma, mesothelioma, ovarian cancer, small cell lung cancer, esophageal cancer, anal cancer, biliary tract cancer, colorectal cancer, cervical cancer, thyroid cancer, salivary gland cancer, prostate cancer, and glioblastoma.

15. The method according to any one of claims 1-13, wherein the cancer is selected from the following: melanoma, non-small cell lung cancer, head and neck cancer, urothelial cancer, breast cancer, gastric cancer, gastroesophageal junction adenocarcinoma, hepatocellular carcinoma, Merkel cell carcinoma, renal cell carcinoma, endometrial cancer, cutaneous squamous cell carcinoma, non-Hodgkin lymphoma, Hodgkin lymphoma, mesothelioma, ovarian cancer, small cell lung cancer, esophageal cancer, colorectal cancer, and cervical cancer.

16. The method according to any one of claims 1-15, wherein the patient has a tumor with a high mutation burden or has a solid tumor with high microsatellite instability (MSI-H) or mismatch repair deficiency.

17. The method according to any one of claims 1-16, wherein the cancer is unresectable or metastatic melanoma; or resected stage IIB, IIC, or III melanoma.

18. The method according to any one of claims 1-16, wherein the cancer is metastatic non-small cell lung cancer (NSCLC).

19. The method according to claim 18, wherein the patient has a tumor with PD-L1 expression measured by a tumor proportion score (TPS) of ≥1% and has not been previously treated with platinum-containing chemotherapy, or the patient has a tumor with PD-L1 expression measured by a tumor proportion score (TPS) of ≥50%.

20. The method according to any one of claims 18-19, wherein the patient's tumor does not have EGFR or ALK genomic abnormalities.

21. The method according to any one of claims 18 - 20, wherein the method further comprises administering to the patient a therapeutically effective amount of pemetrexed and platinum chemotherapy.

22. The method according to claim 21, wherein the patient has non-squamous non-small cell lung cancer, and the pemetrexed is administered to the patient intravenously at a dose of 500 mg / m 2 every 21 days, and the platinum chemotherapy is cisplatin administered to the patient at a dose of 75 mg / m 2 every 21 days.

23. The method according to claim 21 or claim 22, which further comprises administering to the patient about 400 μg to about 1000 μg of folic acid once daily starting about 7 days before administering pemetrexed to the patient and continuing until about 21 days after the last dose of pemetrexed administered to the patient.

24. The method according to any one of claims 21-23, further comprising administering about 1 mg of vitamin B to the patient about 1 week before the first administration of pemetrexed and about every three cycles of pemetrexed administration 12 .

25. The method according to any one of claims 21 - 24, which further comprises administering dexamethasone to the patient twice daily on the day before, on the day of, and on the day after pemetrexed administration.

26. The method according to claim 18, wherein the NSCLC is squamous or non - squamous, and the patient is also treated with a therapeutically effective amount of carboplatin and paclitaxel or nab - paclitaxel.

27. The method according to claim 26, wherein the carboplatin is administered by intravenous infusion at an AUC of 5-6 mg / ml / min, the paclitaxel is administered by intravenous infusion at 200 mg / m every 21 days, 2 and the albumin-bound paclitaxel is administered by intravenous infusion at 100 mg / m every 7 days. 2 administered.

28. The method according to any one of claims 1 - 12, wherein the cancer is resected stage IB, II, or IIIA non - small cell lung cancer.

29. The method according to any one of claims 1 - 12, wherein the cancer is recurrent or metastatic head and neck squamous cell carcinoma (HNSCC) or cervical cancer.

30. The method according to claim 29, wherein the patient's tumor expresses PD - L1 as measured by a combined positive score (CPS) ≥1.

31. The method according to any one of claims 1 - 12, wherein: (1) The patient is an adult and the cancer is recurrent or refractory classical Hodgkin lymphoma (cHL), or (2) the patient is a pediatric patient and the cancer is refractory cHL, or cHL that recurs after 2 or more lines of therapy for cHL.

32. The method according to any one of claims 1 - 12, wherein the cancer is locally advanced or metastatic urothelial carcinoma, locally advanced or metastatic gastric cancer or gastroesophageal junction adenocarcinoma, refractory or recurrent primary mediastinal large B - cell lymphoma (PMBCL), hepatocellular carcinoma, renal cell carcinoma (RCC), recurrent or locally advanced or metastatic Merkel cell carcinoma (MCC).

33. The method according to any one of claims 1 - 12, wherein the cancer is triple - negative breast cancer, ER+ / HER2 - breast cancer, or locally advanced or metastatic esophageal cancer or gastroesophageal junction.

34. The method according to claim 33, wherein the patient's tumor expresses PD - L1 as measured by a combined positive score (CPS) ≥10.

35. The method according to any one of claims 1 - 12, wherein the cancer is advanced renal cell carcinoma (RCC).

36. The method according to any one of claims 1-12, wherein the cancer is selected from the following: melanoma, non-small cell lung cancer, head and neck squamous cell carcinoma, urothelial carcinoma, classical Hodgkin lymphoma, primary mediastinal large B-cell lymphoma (PMBCL), MSI-H cancer, MSI-H or mismatch repair-deficient colorectal cancer, gastric cancer, gastroesophageal junction adenocarcinoma, esophageal cancer, cervical cancer, hepatocellular carcinoma, Merkel cell carcinoma, renal cell carcinoma, endometrial cancer, cutaneous squamous cell carcinoma, tumors with high tumor mutational burden (TMB-H) cancer, and triple-negative breast cancer.

37. The method according to claim 1 and any one of claims 3-36, wherein about 700 units to about 50,000 units of human hyaluronidase are co-formulated with the anti-PD-1 antibody or its antigen-binding fragment.

38. The method according to claim 1 and any one of claims 3-36, wherein the human hyaluronidase is co-formulated with the anti-PD-1 antibody or its antigen-binding fragment at a ratio of about 3 to 36 units: 1 mg.

39. The method according to claim 1 and any one of claims 3-36, wherein the human hyaluronidase is co-formulated with the anti-PD-1 antibody or its antigen-binding fragment at a ratio of about 6 to 25 units: 1 mg.

40. The method according to claim 1 and any one of claims 3-36, wherein the human hyaluronidase is co-formulated with the anti-PD-1 antibody or its antigen-binding fragment at a ratio of about 10 to 14 units: 1 mg.

41. The method according to claim 1 and any one of claims 3-36, wherein the human hyaluronidase is co-formulated with the anti-PD-1 antibody or its antigen-binding fragment at a ratio of about 12.15 units: 1 mg.

42. An anti-PD-1 antibody or its antigen-binding fragment for use in the method of treating cancer according to any one of claims 1-41.

43. An anti-PD-1 antibody for use in the method of treating cancer according to any one of claims 1-41.

44. A pharmaceutical composition for subcutaneous injection comprising an anti-PD-1 antibody or its antigen-binding fragment in a dose of about 600 mg to about 1000 mg and human hyaluronidase, wherein the human hyaluronidase is co-formulated with the anti-PD-1 antibody or its antigen-binding fragment at a ratio of about 3 to 36 units of human hyaluronidase: 1 mg of anti-PD-1 antibody or its binding fragment, wherein the anti-PD-1 antibody or its antigen-binding fragment comprises: a light chain (LC) variable region comprising complementarity-determining regions (CDRs) LC-CDR1, LC-CDR2, and LC-CDR3, wherein LC-CDR1, LC-CDR2, and LC-CDR3 comprise the amino acid sequences shown in SEQ ID NOs: 1, 2, and 3, respectively, and a heavy chain (HC) variable region comprising CDRs HC-CDR1, HC-CDR2, and HC-CDR3, wherein CDRs HC-CDR1, HC-CDR2, and HC-CDR3 comprise the amino acid sequences shown in SEQ ID NOs: 6, 7, and 8, respectively.

45. The pharmaceutical composition according to claim 44, wherein the dose is about 700 mg to about 800 mg of the anti-PD-1 antibody or antigen-binding fragment thereof, and wherein the human hyaluronidase is co-formulated with the anti-PD-1 antibody or antigen-binding fragment thereof at a ratio of about 12.15 units: 1 mg.

46. The pharmaceutical composition according to claim 44, which comprises a 790 mg dose of the anti-PD-1 antibody or antigen-binding fragment thereof and 9600 units of human hyaluronidase.

47. A pharmaceutical composition for subcutaneous injection, which comprises a dose of about 300 mg to about 500 mg of the anti-PD-1 antibody or antigen-binding fragment thereof and human hyaluronidase, wherein the human hyaluronidase is co-formulated with the anti-PD-1 antibody or antigen-binding fragment thereof at a ratio of about 3 to 36 units of human hyaluronidase: 1 mg of anti-PD-1 antibody or antigen-binding fragment thereof, and wherein the anti-PD-1 antibody or antigen-binding fragment thereof comprises: a light chain (LC) variable region comprising complementary determining regions (CDRs) LC-CDR1, LC-CDR2 and LC-CDR3, wherein LC-CDR1, LC-CDR2 and LC-CDR3 comprise the amino acid sequences shown in SEQ ID NOs: 1, 2 and 3 respectively, and a heavy chain (HC) variable region comprising CDR HC-CDR1, HC-CDR2 and HC-CDR3, wherein CDR HC-CDR1, HC-CDR2 and HC-CDR3 comprise the amino acid sequences shown in SEQ ID NOs: 6, 7 and 8 respectively.

48. The pharmaceutical composition according to claim 47, wherein the dose is about 350 mg to about 400 mg of the anti-PD-1 antibody or antigen-binding fragment thereof, and wherein the human hyaluronidase is co-formulated with the anti-PD-1 antibody or antigen-binding fragment thereof at a ratio of about 12.15 units: 1 mg.

49. The pharmaceutical composition according to claim 47, which comprises a 395 mg dose of the anti-PD-1 antibody or antigen-binding fragment thereof and 4800 units of human hyaluronidase.

50. The pharmaceutical composition according to any one of claims 44 to 49, which is in a vial, syringe, infusion device or apparatus.

51. The method, use or pharmaceutical composition according to any one of claims 1 and 3 - 50, wherein the human hyaluronidase is a PH20 variant or a fragment thereof, wherein the PH20 variant has amino acid residue substitutions including M345T, S347T, M348K, K349E, L352Q, L353A, L354I, D355K, N356E, E359D and I361T in SEQ ID NO:16, and the fragment has an N-terminal deletion of amino acid residues 1 - 36, 1 - 37, 1 - 38, 1 - 39, 1 - 40, 1 - 41 or 1 - 42 of SEQ ID NO:16; and / or a C-terminal deletion of amino acid residues 455 - 509, 456 - 509, 457 - 509, 458 - 509, 459 - 509, 460 - 509, 461 - 509, 462 - 509, 463 - 509, 464 - 509, 465 - 509, 466 - 509, 467 - 509, 468 - 509, 469 - 509, 470 - 509, 471 - 509, 472 - 509, 473 - 509, 474 - 509, 475 - 509, 476 - 509, 477 - 509, 478 - 509, 479 - 509, 480 - 509, 481 - 509, 482 - 509, 483 - 509, 484 - 509, 485 - 509, 486 - 509, 487 - 509, 488 - 509, 489 - 509, 490 - 509, 491 - 509, 492 - 509, 493 - 509, 494 - 509, 495 - 509, 496 - 509, 497 - 509, 498 - 509, 499 - 509, 500 - 509, 501 - 509, 502 - 509, 503 - 509, 504 - 509, 505 - 509, 506 - 509, 507 - 509, 508 - 509 or 509, wherein the numbering refers to SEQ ID NO:

16.

52. The method, use or pharmaceutical composition according to any one of claims 1 and 3 - 50, wherein the human hyaluronidase is rHuPH20 or a variant or fragment thereof, and wherein the rHuPH20 or a variant or fragment thereof is SEQ IDAmino acid residues 36 - 464, 36 - 465, 36 - 466, 36 - 467, 36 - 468, 36 - 469, 36 - 470, 36 - 471, 36 - 472, 36 - 473, 36 - 474, 36 - 475, 36 - 476, 36 - 477, 36 - 478, 36 - 479, 36 - 480, 36 - 481, 36 - 482, 36 - 483, 37 - 464, 37 - 465, 37 - 466, 37 - 467, 37 - 468, 37 - 469, 37 - 470, 37 - 471, 37 - 472, 37 - 473, 37 - 474, 37 - 475, 37 - 476, 37 - 477, 37 - 478, 37 - 479, 37 - 480, 37 - 481, 37 - 482, 37 - 483, 38 - 464, 38 - 465, 38 - 466, 38 - 467, 38 - 468, 38 - 469, 38 - 470, 38 - 471, 38 - 472, 38 - 473, 38 - 474, 38 - 475, 38 - 476, 38 - 477, 38 - 478, 38 - 479, 38 - 480, 38 - 481, 38 - 482, 38 - 483, 39 - 464, 39 - 465, 39 - 466, 39 - 467, 39 - 468, 39 - 469, 39 - 470, 39 - 471, 39 - 472, 39 - 473, 39 - 474, 39 - 475, 39 - 476, 39 - 477, 39 - 478, 39 - 479, 39 - 480, 39 - 481, 39 - 482, 39 - 483, 40 - 464, 40 - 465, 40 - 466, 40 - 467, 40 - 468, 40 - 469, 40 - 470, 40 - 471, 40 - 472, 40 - 473, 40 - 474, 40 - 475, 40 - 476, 40 - 477, 40 - 478, 40 - 479, 40 - 480, 40 - 481, 40 - 482, 40 - 483, 41 - 464, 41 - 465, 41 - 466, 41 - 467, 41 - 468, 41 - 469, 41 - 470, 41 - 471, 41 - 472, 41 - 473, 41 - 474, 41 - 475, 41 - 476, 41 - 477, 41 - 478, 41 - 479, 41 - 480, 41 - 481, 41 - 482, 41 - 483, 42 - 464, 42 - 465, 42 - 466, 42 - 467, 42 - 468, 42 - 469, 42 - 470, 42 - 471, 42 - 472, 42 - 473, 42 - 474, 42 - 475, 42 - 476, 42 - 477, 42 - 478, 42 - 479, 42 - 480, 42 - 481, 42 - 482 or 42 - 483.

53. The method, use or pharmaceutical composition according to any one of claims 1 and 3 - 50, wherein the human hyaluronidase is SEQ ID NO:

18.

54. The method, use or pharmaceutical composition according to any one of claims 1 and 3 - 50, wherein the human hyaluronidase is SEQ ID NO:17, 19 or 20.

55. The method, use or pharmaceutical composition according to any one of claims 1 - 54, wherein the anti-PD-1 antibody or an antigen-binding fragment thereof comprises: (a) a heavy chain variable region comprising the amino acid sequence as shown in SEQ ID NO:9 or a variant of SEQ ID NO:9, and (b) a light chain variable region comprising the amino acid sequence as shown in SEQ ID NO:4 or a variant of SEQ ID NO:

4.

56. The method, use or pharmaceutical composition according to any one of claims 1-54, wherein the anti-PD-1 antibody or antigen-binding fragment thereof comprises a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO:9, and a light chain variable region comprising the amino acid sequence shown in SEQ ID NO:

4.

57. The method, use or pharmaceutical composition according to any one of claims 1-42 and 44-56, wherein the anti-PD-1 antibody or antigen-binding fragment thereof is an anti-PD-1 antibody.

58. The method, use or pharmaceutical composition according to claim 57, wherein the anti-PD-1 antibody is a monoclonal antibody comprising: (a) a heavy chain comprising the amino acid sequence shown in any one of SEQ ID NO:10-15 or a variant of any one of SEQ ID NO:10-15, and (b) a light chain comprising the amino acid sequence shown in SEQ ID NO:5 or a variant of SEQ ID NO:

5.

59. The method, use or pharmaceutical composition according to claim 58, wherein the anti-PD-1 antibody is a monoclonal antibody comprising a heavy chain consisting of the amino acid sequence shown in SEQ ID NO:10 and a light chain consisting of the amino acid sequence shown in SEQ ID NO:

5.

60. The method, use or pharmaceutical composition according to claim 58, wherein the anti-PD-1 antibody is a monoclonal antibody comprising a heavy chain consisting of the amino acid sequence shown in SEQ ID NO:11 and a light chain consisting of the amino acid sequence shown in SEQ ID NO:

5.

61. The method, use or pharmaceutical composition according to claim 58, wherein the anti-PD-1 antibody is pembrolizumab.

62. The method, use or pharmaceutical composition according to claim 58, wherein the anti-PD-1 antibody is a pembrolizumab variant.

63. The method, use or pharmaceutical composition according to any one of claims 1-62, wherein the anti-PD-1 antibody or antigen-binding fragment thereof is in a composition comprising 130 mg / mL of the anti-PD-1 antibody or antigen-binding fragment thereof.

64. The method, use or pharmaceutical composition according to any one of claims 1-62, wherein the anti-PD-1 antibody or antigen-binding fragment thereof is in a composition comprising 165 mg / mL of the anti-PD-1 antibody or antigen-binding fragment thereof.

65. The method, use or pharmaceutical composition according to claim 63 or claim 64, wherein the composition comprises 500 to 8000 U / ml of the human hyaluronidase.

66. The method, use or pharmaceutical composition according to claim 63 or claim 64, wherein the composition comprises 2000 U / ml of the human hyaluronidase.

67. The method, use or pharmaceutical composition according to any one of claims 63 to 66, wherein the composition further comprises 10 mM L-methionine, 10 mM histidine, pH 5.5, 7% sucrose and 0.02% polysorbate 80.

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