Aqueous pharmaceutical compositions of anti-PD1 antibody prorugolimab and uses thereof

Abstract

The present invention relates to aqueous pharmaceutical compositions for the anti-PD-1 antibody prorugolimab, and the use of such pharmaceutical compositions as pharmaceutical agents for treating PD-1 mediated diseases.

Description

[Technical field]

[0001]

[0001] The present invention relates to a novel aqueous composition for an anti-PD-1 antibody, in particular, a novel aqueous composition for the anti-PD-1 antibody prorugolimab, which can be used as a medicinal agent for treating malignant neoplasms. [Background technology]

[0002]

[0002] The programmed cell death 1 (PD-1) protein is an inhibitory member of the CD28 receptor family, which also includes CD28, CTLA-4, ICOS and BTLA. PD-1 is expressed by activated B cells, T cells and myeloid cells (Agata et al., supra; Okazaki et al. (2002) Curr. Opin. Immunol. 14:391779-82; Bennet et al. (2003) J Immunol 170:711-8). The first members of this family, CD28 and ICOS, were detected by their functional effect on increasing T cell proliferation after the addition of monoclonal antibodies (Hutloff et al. (1999) Nature 397:263-266; Hansen et al. (1980) Immunogenics 10:247-260). PD-1 was discovered by screening for differential expression in apoptotic cells (Ishida et al. (1992) EMBO J 11:3887-95). Other members of the family, CTLA-4 and BTLA, were discovered by screening for differential expression in cytotoxic T lymphocytes and TH1 cells, respectively. CD28, ICOS, and CTLA-4 all have unpaired cysteine ​​residues that allow homodimerization. In contrast, PD-1 exists as a monomer and appears to lack the unpaired cysteine ​​residues characteristic of other members of the CD28 family.

[0003] PD-1 is a 55 kDa type I transmembrane protein that is a member of the Ig gene superfamily (Agata et al. (1996) Int Immunol 8:765-72). PD-1 contains a membrane-proximal immunoreceptor tyrosine-based inhibitory motif (ITIM) and a membrane-distal tyrosine-based switch motif (ITSM) (Thomas, ML (1995) J Exp Med 181:1953-6; Vivier, E and Daeron, M (1997) Immunol Today 18:286-91). Although structurally similar to CTLA-4, PD-1 lacks the MYPPY motif that is important for B7-1 and B7-2 binding. PD-1 has been detected to have two ligands, PD-L1 and PD-L2, and has been shown to negatively regulate T cell activation after binding to PD-1 (Freeman et al. (2000) J Exp Med 192:1027-34; Latchman et al. (2001) Nat Immunol 2:261-8; Carter et al. (2002) Eur J Immunol 32:634-43). Both PD-L1 and PD-L2 are B7 homologs that bind to PD-1 but not other members of the CD28 family.

[0004]

[0004] PD-L1, one of the PD-1 ligands, is abundant in various types of human cancers (Dong et al. (2002) Nat. Med. 8:787-9). The interaction between PD-1 and PD-L1 results in a reduction in the number of tumor-infiltrating lymphocytes, a decrease in T cell receptor-mediated proliferation, and the escape of cancer cells from immune surveillance (Dong et al. (2003) J. Mol. Med. 81:281-7; Blank et al. (2005) Cancer Immunol. Immunother. 54:307-314; Konishi et al. (2004) Clin. Cancer Res. 10:5094-100). Immune suppression can be reversed by inhibiting the local PD-L1 / PD-1 interaction, and this effect is additive when the PD-L2 / PD-1 interaction is blocked (Iwai et al. (2002) Proc. Nat'l. Acad. Sci. USA 99:12293-7; Brown et al. (2003) J. Immunol. 170:1257-66).

[0005] PD-1 is an inhibitory member of the CD28 family and is expressed on activated B cells, T cells, and myeloid cells (Agata et al., supra; Okazaki et al. (2002) Curr Opin Immunol 14:391-779-82; Bennett et al. (2003) J Immunol 170:711-8). PD-1-deficient animals are prone to develop a variety of autoimmune diseases, including autoimmune heart disease, and lupus-like syndromes, including arthritis and nephritis (Nishimura et al. (1999) Immunity 11:141-51; Nishimura et al. (2001) Science 291:319-22). Furthermore, PD-1 has been found to play a role in autoimmune encephalomyelitis, systemic lupus erythematosus, graft-versus-host disease (GVHD), type I diabetes, and rheumatoid arthritis (Salama et al. (2003) J Exp Med 198:71-78; Prokunina and Alarcon-Riquelme (2004) Hum Mol Genet 13:R143; Nielsen et al. (2004) Lupus 13:510). In mouse B-cell tumor lines, the ITSM of PD-1 was shown to be essential for blocking BCR-mediated Ca2+ flux and tyrosine phosphorylation of downstream effector molecules (Okazaki et al. (2001) PNAS 98:13866-71).

[0006]

[0006] Several anti-PD-1 antibodies are known in the art, such as nivolumab (BMS) and pembrolizumab (Merck), which are human IgG4 monoclonal antibodies.

[0007]

[0007] Also known is the novel anti-PD1 antibody prorugolimab (also known as BCD-100), a monoclonal human antibody of IgG1 isotype non-effector mutations L234A, L235A. Prorugolimab has been shown to have increased affinity for PD-1 and increased aggregation stability compared to IgG4 antibodies. Furthermore, prorugolimab is currently in clinical trials for various types of malignant neoplasms, including inoperable or metastatic melanoma, melanoma including early-stage melanoma before and after definitive treatment, lung cancer, and non-small cell lung cancer including inoperable or metastatic non-small cell lung cancer (NSCLC).

[0008]

[0008] In light of the above, it is pertinent to create new and improved stable aqueous pharmaceutical compositions for the anti-PD-1 antibody prorugolimab.

[0009] The present invention will be better understood from the following detailed description of embodiments of the invention, taken in conjunction with the accompanying drawings, in which: [Brief description of the drawings]

[0009] [Figure 1]

[0010] FIG. 1 shows the dynamics of BCD-100 concentration μg / ml in patient serum over six doses (adjusted by factors) (BCD-100-1 study). [Diagram 2] FIG. 1 shows the dynamics of BCD-100 concentration μg / ml in patient serum over six doses (adjusted by factors) (BCD-100-1 study). [Diagram 3]

[0011] FIG. 1 shows the BCD-100-2 / MIRACULUM trial design. [Figure 4]

[0012] FIG. 1 shows a schematic diagram of the test. [Diagram 5]

[0013] FIG. 1 shows the overall survival rate of patients in group 1 (BCD-100, 1 mg / kg every 2 weeks) according to the results of the BCD-100-2 / MIRACULUM trial. [Figure 6]

[0014] FIG. 1 shows the overall survival rate of patients in group 2 (BCD-100, 3 mg / kg every 3 weeks) according to the results of the BCD-100-2 / MIRACULUM trial. [Figure 7]

[0015] FIG. 1 shows the progression-free survival rate (based on irRECIST criteria) of patients in the BCD-100 1 mg / kg group according to the results of the BCD-100-2 / MIRACULUM study. [Figure 8]

[0016] FIG. 1 shows the progression-free survival rate (based on irRECIST criteria) of patients in the BCD-100 3 mg / kg group according to the results of the BCD-100-2 / MIRACULUM study. [Figure 9]

[0017] FIG. 1 shows BCD-100 concentrations in patients receiving 1 mg / kg every 2 weeks after administration of a single dose of the product (results from the BCD-100-2 / MIRACULUM study). [Figure 10]

[0018] FIG. 1 shows BCD-100 concentrations in patients receiving 3 mg / kg every 3 weeks (results from the BCD-100-2 / MIRACULUM study). [Figure 11]

[0019] FIG. 1 shows the proportion of Th9 in the total population of helper T cells in a subgroup of patients who received BCD-100 at a dose of 1 mg / kg Q2W, indicating different types of responses to treatment (results from the BCD-100-2 / MIRACULUM study). [Figure 12]

[0020] FIG. 1 shows the proportion of Th9 in the total population of helper T cells in a subgroup of patients who received BCD-100 at a dose of 3 mg / kg Q3W, indicating different types of responses to treatment (results from the BCD-100-2 / MIRACULUM study). DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0010] definition

[0021] The terms used herein generally have their ordinary meanings in the art, in the context of the present invention, and in the specific context in which each term is used. Certain terms used to describe the present invention are discussed below or elsewhere in this specification to provide additional guidance to practitioners regarding the description of the present invention. Synonyms of certain terms are provided. The specification of one or more synonymous terms does not exclude the use of other synonymous terms. The use of examples anywhere in this specification, including examples of any term discussed herein, is merely illustrative and does not limit the scope and meaning of the present invention or any exemplary term in any way. The present invention is not limited to the various aspects provided herein.

[0011]

[0022] "Monoclonal antibody" as used herein relates to a humanized or fully human antibody, unless otherwise stated in the application. The monoclonal antibody according to the invention can be obtained, for example, using recombinant techniques, phage display techniques, synthetic techniques or a combination thereof, or other techniques well known from the prior art.

[0012]

[0023] A population of "monoclonal antibodies" as used herein refers to a homogenous or essentially homogenous antibody population (i.e., at least 96% or 96%, more preferably about 97% or more or 98% or more, or even more preferably at least 99% of the antibodies in the population compete for the same antigen / epitope in an enzyme-linked immunosorbent assay ELISA, or even more preferably the antibodies are identical in amino acid sequence).

[0013]

[0024] Natural full-size antibodies are represented by immunoglobulin molecules that contain four polypeptide chains (two heavy (H) chains with total length of about 50-70 KDa, and two light (L) chains with total length of about 25 KDa) linked through disulfide bonds. The amino-terminal portion of each chain contains a variable domain of about 100-110 or more amino acids involved in binding to the antigen. The carboxyl-terminal domain of each chain determines the constant region that is mostly responsible for effector functions. Light chains are classified as kappa and lambda and are characterized by specific constant regions. Each light chain is characterized by containing a variable N-terminal light chain region (hereafter referred to as VL or VK), and a constant light chain region (CL or CK) consisting of a single domain. Heavy chains are classified as gamma, delta, alpha, mu, and epsilon, which define the immunoglobulin classes IgG, IgM, IgA, IgD, and IgE, respectively. Some of these can be further divided into subclasses (isotypes), such as IgG1, IgG2, IgG3, IgG4, IgA1 and IgA2. Each heavy chain type is characterized by a specific constant region Fc. Each heavy chain comprises a variable N-terminal region (hereafter referred to as VH) and a constant region CH. The constant heavy chain region consists of three domains (CH1, CH2 and CH3) for IgG, IgD and IgA, and four domains (CH1, CH2, CH3 and CH4) for IgM and IgE. The VH and VL variable domains can also be divided into so-called hypervariable regions (complementarity determining regions, CDRs) interspersed with more conserved framework regions (FRs). Each variable domain comprises three CDRs and four FRs, located in the following order from N-terminus to C-terminus: FR1, CDR1, FR2, CDR2, FR3, CDR3 and FR4.

[0014]

[0025] The variable regions of each light / heavy chain pair form the antigen-binding site of the antibody. Thus, an intact IgG antibody has two binding sites. Except for bifunctional or bispecific antibodies, the two binding sites are identical. As used herein, "antigen-binding portion" or "antigen-binding region" or "antigen-binding domain" are interchangeable with the region of an antibody that contains the amino acid residues that interact with the antigen and confer to the antibody its specificity and affinity for the antigen. This antibody fragment contains the frame amino acid residues necessary to maintain the proper conformation of the antigen-binding residues.

[0015]

[0026] "Antibody fragment" can be represented by an antibody fragment having the activity of a full-sized antibody. The antibody fragment can be represented by F(ab')2, F(ab)2, Fab', Fab Fv and scFv.

[0016]

[0027] As used herein, the terms "inhibit" or "neutralize" with respect to the activity of an antibody of the invention refer to the ability to significantly block, prevent, limit, slow, stop, reduce, or reverse, for example, the occurrence or severity of the biological activity (such as the activity of PD-1) or trait, disease, or condition that is being inhibited, including but not limited to, the activity of PD-1. Inhibition or neutralization of PD-1 activity resulting from binding of an antibody of the invention to PD-1 is preferably at least about 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95% or more.

[0017]

[0028] The terms "isolated" or "isolated" with respect to a nucleic acid or protein product (e.g., an antibody) refer to a nucleic acid or protein molecule that is identified and separated from at least one of the contaminants with which it is ordinarily associated in a natural source. Preferably, an "isolated antibody" is an antibody that is substantially free of other antibodies having a particular antigen specificity (e.g., a pharmaceutical composition of the invention contains an isolated antibody that specifically binds to PD-1 and is substantially free of antibodies that specifically bind to antigens other than PD-1).

[0018]

[0029] A polynucleotide is "operably linked" when it has a functional link with another polynucleotide. For example, a promoter or enhancer is operably linked to a coding sequence if it affects the transcription of the sequence. A polypeptide is "operably linked" to another polypeptide when the polynucleotide that codes for it is operably linked, preferably when they are located in the same open reading frame.

[0019]

[0030] The term "specific binding" between an antibody and an antigen target (antigen) refers to immunological specificity. An antibody can specifically bind to an antigen target if it binds to the antigen epitope stronger than other antigen epitopes. Specific binding does not exclude cross-reactivity with other antigens that have similar antigen epitopes.

[0020]

[0031] The VL domain in the antibody of the present invention can be either of the VLlambda or VLkappa type. The term "VL domain" encompasses both VLlambda and VLkappa isotypes that contain one or more amino acid substitutions, insertions or deletions.

[0021]

[0032] The term "pharmaceutical composition" refers to a composition and / or formulation containing a therapeutically effective amount of an antibody of the invention and excipients (carriers, diluents, vehicles, solvents and other excipients).

[0033] The term "buffer" or "buffer solution" as used herein refers to an aqueous solution containing a mixture of an acid (typically a weak acid, e.g., acetic acid, citric acid, etc.) and its conjugate base (e.g., acetate or citrate salts, e.g., sodium acetate, sodium citrate, and hydrates of the above salts, e.g., sodium acetate trihydrate, etc.), or alternatively, a mixture of a base (typically a weak base, e.g., histidine) and its conjugate acid (e.g., histidine hydrochloride). The pH of a "buffer solution" changes only slightly with the addition of small amounts of a strong base or strong acid, as well as dilution or concentration due to the "buffering effect" imparted by the "buffering agent."

[0022]

[0034] As used herein, a "buffer system" includes one or more buffering agents and / or acid / base conjugates thereof, more suitably one or more buffering agents and their acid / base conjugates, and most suitably one buffering agent and its acid / base conjugates. Unless otherwise specified, any concentration referred to herein as a "buffer system" (buffer concentration) can suitably refer to the combined concentration of the buffering agent(s) and / or its acid / base conjugate(s). In other words, a concentration referred to herein as a "buffer system" can refer to the combined concentration of all relevant buffer species (i.e., species that are in dynamic equilibrium with each other, e.g., citrate / citric acid). The overall pH of a composition that includes an associated buffer system is a reflection of the equilibrium concentration of each of the associated buffering species (i.e., the balance of the buffering agent(s) and their acid / base conjugates).

[0023]

[0035] The term "buffering agent" as used herein refers to the acid or base component (typically a weak acid or weak base) of a buffer or buffer solution. A buffering agent helps maintain the pH of a given solution at or near a given value, and the buffering agent is generally selected to complement the given value. A buffering agent may be a single compound that produces the desired buffering effect, especially when the buffering agent is mixed with (and is appropriately proton exchangeable with) an appropriate amount of its corresponding "acid / base conjugate" (depending on the desired given pH).

[0024]

[0036] As used herein, the term "solubilizer" refers to a pharma-ceutically acceptable non-ionic surfactant. Both one solubilizer and their combinations can be used. Exemplary solubilizers include, but are not limited to, polysorbate 20 or polysorbate 80, poloxamer 184 or poloxamer 188, or PLURONIC®.

[0025]

[0037] The terms "osmotic agent" or "tonicity agent", as well as "osmolyte", as used herein refer to an excipient capable of providing the required osmolarity of a liquid antibody solution. In some embodiments, the tonicity agent can increase the osmolarity of a liquid antibody formulation to isotonicity so that the liquid antibody formulation is physiologically compatible with the cells of the tissue of the subject organism. In another embodiment, the tonicity agent can contribute to increasing the stability of the antibody. An "isotonic" drug is a drug that has an osmolarity equivalent to that of human blood. An isotonic drug typically has an osmolarity of about 250-350 mOsm / kg. The term "hypotonic" describes a formulation that has an osmolarity lower than that of human blood. Correspondingly, the term "hypertonic" is used to describe a formulation that has an osmolarity higher than that of human blood. Isotonicity can be measured, for example, using a vapor pressure or freezing point depression osmometer. The tonicity agent may be in an enantiomeric form (e.g., L- or D-enantiomer) or racemic form; in an isomeric form, such as alpha or beta, e.g., alpha, alpha; or beta, beta; or alpha, beta; or beta, alpha; in a free acid or free base form; in a salt form; in a hydrated form (e.g., monohydrate) or anhydrous form. Exemplary osmotic agents include, but are not limited to, sugars (trehalose dihydrate, sucrose, glucose), polyols (mannitol, sorbitol), amino acids (proline, arginine, glycine), or salts (sodium chloride, potassium chloride, magnesium chloride).

[0026]

[0038] The term "long-term storage" or "long-term stability" is understood to mean that the pharmaceutical composition may be stored with a minimum stable shelf-life of 3 months or more, 6 months or more, preferably 1 year or more, and most preferably at least 2 years. In general, the terms "long-term storage" and "long-term stability" further include a stable shelf-life that is at least equivalent to or better than the stable shelf-life typically required for currently available commercial formulations of the anti-PD-1 antibody prorugolimab, without compromising stability that would render the formulation unsuitable for its intended pharmaceutical use.

[0027]

[0039] The term "parenteral administration" typically refers to administration regimens by injection and includes intravenous, intramuscular, intraarterial, intratracheal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, intratracheal, subcutaneous, intraarticular, subcapsular, subarachnoid, intraspinal, epidural and intrasternal injection or infusion, among others.

[0028]

[0040] The term "use" refers to the ability to use the antibody of the present invention or a pharmaceutical composition containing it to treat disease, ameliorate the course of disease, promote remission, or reduce the recurrence rate of a disease or disorder mediated by a receptor to which the antibody of the present invention can bind. Exemplary diseases include, but are not limited to, malignant neoplasms, including inoperable or metastatic melanoma, melanoma, including early stage melanoma before and after definitive treatment; lung cancer, non-small cell lung cancer, including inoperable or metastatic non-small cell lung cancer (NSCLC); non-squamous non-small cell lung cancer, squamous cell lung cancer; small cell lung cancer, including inoperable or metastatic small cell lung cancer; early stage lung cancer before and after definitive treatment; metastatic cervical cancer, including early stage cervical cancer before and after definitive treatment; head and neck tumors, including head and neck squamous cell carcinoma; Hodgkin's lymphoma; gastric and intestinal tumours, metastatic squamous cell esophageal cancer; bladder cancer including metastatic urothelial carcinoma, renal cancer; metastatic endometrial cancer, including early stage endometrial cancer before and after definitive treatment; metastatic breast cancer, including early stage endometrial cancer before and after definitive treatment; metastatic or inoperable liver cancer, including early stage liver cancer before and after definitive treatment; inoperable or metastatic solid tumours including inoperable or metastatic solid tumours with signs of microsatellite instability.

[0029]

[0041] The term "therapeutic method" refers to the ability to treat disease, ameliorate the course of disease, promote remission, or reduce the recurrence rate of disease or disorder associated with PD1 activity using the antibody of the present invention or a pharmaceutical composition containing it. "Treating" or "treatment" of a disease, disorder, or condition can include preventing or delaying the onset of clinical symptoms of the disease, disorder, or condition occurring in humans, inhibiting the disease, disorder, or condition, i.e., arresting, reducing, or delaying the onset of the disease or its recurrence (in the case of maintenance therapy) or at least one clinical or subclinical symptom thereof, or alleviating or relieving the disease, i.e., causing the regression of the disease, disorder, or condition. Exemplary diseases are, but are not limited to, malignant neoplasms, such as melanoma, including inoperable or metastatic melanoma, early stage melanoma before or after definitive surgical treatment; lung cancer, non-small cell lung cancer, including inoperable or metastatic non-small cell lung cancer (NSCLC).

[0030]

[0042] As used herein, the term "aqueous composition" refers to a water-based composition, and the water in the composition may be water, water for injection, or saline (a 0.9% to 1.0% aqueous solution of sodium chloride).

[0031]

[0043] In one aspect of the invention, the subject, or patient, of treatment is a mammal, preferably a human subject. The subject may be male or female of any age.

[0044] As used herein, the terms "comprise," "having," "include," or variations such as "comprises," "comprising," "has," "having," "includes," or "including," and all grammatical variations thereof, are understood to mean the inclusion of a stated integer or group of integers but not the exclusion of any other integer or group of integers. Summary of the Invention [Means for solving the problem]

[0032]

[0045] The present invention discloses an aqueous pharmaceutical composition of the anti-PD-1 antibody prorugolimab, which has improved aggregation stability and increased affinity compared to known anti-PD-1 antibodies based on human antibodies of the IgG4 isotype.

[0033]

[0046] As disclosed in International Patent Application Publication No. 2018 / 013017, which is incorporated herein by reference, the anti-PD-1 antibody prorugolimab (herein referred to as the "antibody of the invention"), a monoclonal human antibody of IgG1 isotype with non-effector mutations L234A, L235A, has been shown to have improved aggregation stability, increased affinity, and improved pharmacokinetic parameters, such as t1 / 2β (hours) or Cmax (μg / ml), when compared to known anti-PD-1 antibodies based on human antibodies of IgG4 isotype, such as nivolumab. Prorugolimab has a weight-average molecular weight of about 146 kDa and is specific for human PD-1. Prorugolimab has a heavy chain containing 459 amino acids (SEQ ID NO: 1) and a human light chain containing 214 amino acids (SEQ ID NO: 2), and the constant portion (Fc) of prorugolimab contains the L234A, L235A mutations.

[0034]

[0047] It may therefore be advantageous to administer an aqueous composition together with the antibodies of the present invention to a patient with a malignant neoplasm.

[0048] A broad aspect of the invention is an aqueous pharmaceutical composition suitable for administration to a subject to inhibit PD-1 protein activity, the aqueous pharmaceutical composition comprising a pharma- ceutical effective amount of the anti-PD-1 antibody prorugolimab, and an effective amount of trehalose dihydrate, acetate, or a histidine-based buffer.

[0035]

[0049] According to one broad aspect of the invention, (a) Prorugolimab at a concentration of 15 mg / ml to 40 mg / ml as an antibody; (b) trehalose dihydrate at a concentration of 80 mg / ml to 110 mg / ml; (c) sodium acetate trihydrate at a concentration between 0.2 mg / mL and 2.5 mg / mL; and (d) Acetic acid in an amount that makes the pH 4.5 to 5.5 and an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

[0036]

[0050] In some aspects of the invention, the prorugolimab may be present at a concentration of 15 mg / ml to 25 mg / ml.

[0051] In some aspects of the invention, the prorugolimab may be present at a concentration of 20 mg / ml.

[0037]

[0052] In some embodiments of the invention, the trehalose dihydrate may be present in a concentration of from 95 mg / ml to 105 mg / ml.

[0053] In some aspects of the invention, the trehalose dihydrate may be present at a concentration of 100 mg / ml.

[0038]

[0054] In some aspects of the invention, the sodium acetate trihydrate may be present in a concentration of 1.6 mg / ml to 1.9 mg / ml.

[0055] In some aspects of the invention, the sodium acetate trihydrate may be present in a concentration of 1.7 mg / ml to 1.8 mg / ml.

[0039]

[0056] In some aspects of the invention, the sodium acetate trihydrate may be present at a concentration of 1.742 mg / ml.

[0057] In some embodiments of the invention, the acetic acid may be added to bring the pH to 5.0.

[0040]

[0058] In some aspects of the invention, the acetic acid may be present in a concentration of 0.04 mg / ml to 0.77 mg / ml.

[0059] In some aspects of the invention, the acetic acid may be present at a concentration of 0.40 mg / ml to 0.50 mg / ml.

[0041]

[0060] In some aspects of the invention, the acetic acid may be present at a concentration of 0.43 mg / ml.

[0061] According to one broad aspect of the invention, (a) Prorugolimab at a concentration of 90 mg / ml to 150 mg / ml as an antibody; (b) trehalose dihydrate at a concentration of 50 mg / ml to 110 mg / ml; (c) sodium acetate trihydrate at a concentration between 0.2 mg / mL and 2.5 mg / mL; and (d) Acetic acid in an amount that makes the pH 4.5 to 5.5 and an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

[0042]

[0062] In some embodiments of the invention, the prorugolimab may be present at a concentration of 90 mg / ml to 110 mg / ml.

[0063] In some aspects of the invention, the prorugolimab may be present at a concentration of 100 mg / ml.

[0043]

[0064] In some embodiments of the invention, the trehalose dihydrate may be present in a concentration of 75 mg / ml to 85 mg / ml.

[0065] In some aspects of the invention, the trehalose dihydrate may be present at a concentration of 80 mg / ml.

[0044]

[0066] In some aspects of the invention, the sodium acetate trihydrate may be present in a concentration of 1.6 mg / ml to 1.9 mg / ml.

[0067] In some aspects of the invention, the sodium acetate trihydrate may be present in a concentration of 1.7 mg / ml to 1.8 mg / ml.

[0045]

[0068] In some aspects of the invention, the sodium acetate trihydrate may be present at a concentration of 1.742 mg / ml.

[0069] In some embodiments of the present invention, the pH can be adjusted to 5.0 to 5.5 by adding the above-mentioned acetic acid.

[0046]

[0070] In some embodiments of the invention, the acetic acid may be added to bring the pH to 5.0.

[0071] In some aspects of the invention, the acetic acid may be present in a concentration of 0.045 mg / ml to 0.77 mg / ml.

[0047]

[0072] In some aspects of the invention, the acetic acid may be present at a concentration of 0.40 mg / ml to 0.50 mg / ml.

[0073] In some aspects of the invention, the acetic acid may be present at a concentration of 0.43 mg / ml.

[0048]

[0074] According to one broad aspect of the invention, (a) Prorugolimab as an antibody at a concentration of 5 mg / ml to 150 mg / ml; (b) trehalose dihydrate at a concentration of 70 mg / ml to 110 mg / ml; (с) L-histidine in a concentration of 0.2 to 2.5 mg / ml; and (d) L-histidine hydrochloride at a concentration of 0.2 to 3.5 mg / ml and an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

[0049]

[0075] In some aspects of the invention, the prorugolimab may be present at a concentration of 15 mg / ml to 40 mg / ml.

[0076] In some aspects of the invention, the prorugolimab may be present at a concentration of 15 mg / ml to 25 mg / ml.

[0050]

[0077] In some aspects of the invention, the prorugolimab may be present at a concentration of 20 mg / ml.

[0078] In some embodiments of the invention, the trehalose dihydrate may be present in a concentration of from 95 mg / ml to 105 mg / ml.

[0051]

[0079] In some aspects of the invention, the trehalose dihydrate may be present at a concentration of 100 mg / ml.

[0080] In some embodiments of the present invention, the L-histidine may be present at a concentration of 0.7 mg / ml to 1.0 mg / ml.

[0052]

[0081] In some aspects of the invention, the L-histidine may be present at a concentration of 0.92 mg / ml.

[0082] In some aspects of the invention, the L-histidine hydrochloride may be present at a concentration of 2.8 mg / ml to 3.3 mg / ml.

[0053]

[0083] In some aspects of the invention, the L-histidine hydrochloride may be present at a concentration of 2.96 mg / ml.

[0084] In some aspects of the invention, the composition may have a pH of 5.5 to 6.5.

[0054]

[0085] In some aspects of the invention, the composition may have a pH of 5.5.

[0086] In some embodiments of the invention, the prorugolimab may be present at a concentration of 90 mg / ml to 110 mg / ml.

[0055]

[0087] In some aspects of the invention, the prorugolimab may be present at a concentration of 100 mg / ml.

[0088] In some embodiments of the invention, the trehalose dihydrate may be present in a concentration of 75 mg / ml to 85 mg / ml.

[0056]

[0089] In some aspects of the invention, the trehalose dihydrate may be present at a concentration of 80 mg / ml.

[0090] In some embodiments of the present invention, the L-histidine may be present at a concentration of 0.7 mg / ml to 1.0 mg / ml.

[0057]

[0091] In some aspects of the invention, the L-histidine may be present at a concentration of 0.92 mg / ml.

[0092] In some aspects of the invention, the L-histidine hydrochloride may be present at a concentration of 2.8 mg / ml to 3.3 mg / ml.

[0058]

[0093] In some aspects of the invention, the L-histidine hydrochloride may be present at a concentration of 2.96 mg / ml.

[0094] In some aspects of the invention, the composition may have a pH of 5.5 to 6.5.

[0059]

[0095] In some aspects of the invention, the composition may have a pH of 5.5 to 6.0.

[0096] In some aspects of the invention, the composition may have a pH of 5.5.

[0097] The aqueous pharmaceutical composition of the anti-PD-1 antibody prorugolimab of the invention may further comprise a suitable solubilizing agent.

[0060]

[0098] In some embodiments of the present invention, the solubilizing agent may be poloxamer 188.

[0099] In some embodiments of the invention, the poloxamer 188 may be present in an amount greater than 0 mg / ml but less than or equal to 1 mg / ml.

[0061]

[0100] In some embodiments of the invention, the poloxamer 188 may be present in amounts of 0 mg / ml, 0.1 mg / ml, 0.2 mg / ml, 0.3 mg / ml, 0.4 mg / ml, 0.5 mg / ml, 0.6 mg / ml, 0.7 mg / ml, 0.8 mg / ml, 0.9 mg / ml, 1.0 mg / ml.

[0062]

[0101] In one aspect, the present invention provides a method for producing a method for treating a cancer cell comprising: (a) Prorugolimab at a concentration of 20 mg / ml as antibody; (b) trehalose dihydrate at a concentration of 100 mg / ml; (с) sodium acetate trihydrate in a concentration of 0.2 to 2.5 mg / ml; and (d) Acetic acid in an amount that makes the pH 4.5 to 5.5 The present invention relates to an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising the compound.

[0063]

[0102] In one aspect, the present invention provides a method for producing a method for treating a cancer cell comprising: (a) Prorugolimab at a concentration of 20 mg / ml as antibody; (b) trehalose dihydrate at a concentration of 100 mg / ml; (с) sodium acetate trihydrate in a concentration of 1.7 mg / ml to 1.8 mg / ml; and (d) Acetic acid in an amount that makes the pH 5.0 The present invention relates to an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising the compound.

[0064]

[0103] In one aspect, the present invention provides a method for producing a method for treating a cancer cell comprising: (a) Prorugolimab at a concentration of 20 mg / ml as antibody; (b) trehalose dihydrate at a concentration of 100 mg / ml; (с) sodium acetate trihydrate in a concentration of 1.742 mg / ml; and (d) Acetic acid in an amount that makes the pH 5.0 The present invention relates to an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising the compound.

[0065]

[0104] In one aspect, the present invention provides a method for producing a method for treating a cancer cell comprising: (a) Prorugolimab at a concentration of 100 mg / ml as antibody; (b) trehalose dihydrate at a concentration of 80 mg / ml; (c) sodium acetate trihydrate in a concentration of 0.2 to 2.5 mg / ml; and (d) Acetic acid in an amount that makes the pH 4.5 to 5.5 The present invention relates to an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising the compound.

[0066]

[0105] In one aspect, the present invention provides a method for producing a method for treating a cancer cell comprising: (a) Prorugolimab at a concentration of 100 mg / ml as antibody; (b) trehalose dihydrate at a concentration of 80 mg / ml; (с) sodium acetate trihydrate in a concentration of 1.7 mg / ml to 1.8 mg / ml; and (d) Acetic acid in an amount that makes the pH 5.0 to 5.5 The present invention relates to an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising the compound.

[0067]

[0106] In one aspect, the present invention provides a method for producing a method for treating a cancer cell comprising: (a) Prorugolimab at a concentration of 100 mg / ml as antibody; (b) trehalose dihydrate at a concentration of 80 mg / ml; (с) sodium acetate trihydrate in a concentration of 1.742 mg / ml; and (d) Acetic acid in an amount that makes the pH 5.0 to 5.5. The present invention relates to an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising the compound.

[0068]

[0107] In one aspect, the present invention provides a method for producing a method for treating a cancer cell comprising: (a) Prorugolimab at a concentration of 100 mg / ml as antibody; (b) trehalose dihydrate at a concentration of 80 mg / ml; (с) sodium acetate trihydrate in a concentration of 1.742 mg / ml; and (d) Acetic acid in an amount that makes the pH 5.0 The present invention relates to an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising the compound.

[0069]

[0108] In one aspect, the invention provides an aqueous pharmaceutical composition of an anti-PD-1 antibody, comprising: (a) Prorugolimab at a concentration of 20 mg / ml as antibody; (b) trehalose dihydrate at a concentration of 100 mg / ml; (с) L-histidine in a concentration of 0.2 to 2.5 mg / ml; and (d) L-histidine hydrochloride at a concentration of 0.2 to 3.5 mg / ml Including, (e) The above composition relates to an aqueous pharmaceutical composition having a pH of 5.5 to 6.5.

[0070]

[0109] In one aspect, the invention provides an aqueous pharmaceutical composition of an anti-PD-1 antibody, comprising: (a) Prorugolimab at a concentration of 20 mg / ml as antibody; (b) trehalose dihydrate at a concentration of 100 mg / ml; (с) L-histidine at a concentration of 0.92 mg / ml; and (d) L-histidine hydrochloride at a concentration of 2.96 mg / ml Including, (e) The above composition relates to an aqueous pharmaceutical composition having a pH of 5.5.

[0071]

[0110] In one aspect, the invention provides an aqueous pharmaceutical composition of an anti-PD-1 antibody, comprising: (a) Prorugolimab at a concentration of 100 mg / ml as antibody; (b) trehalose dihydrate at a concentration of 80 mg / ml; (с) L-histidine in a concentration of 0.2 to 2.5 mg / ml; and (d) L-histidine hydrochloride at a concentration of 0.2 to 3.5 mg / ml Including, (e) The above composition relates to an aqueous pharmaceutical composition having a pH of 5.5 to 6.5.

[0072]

[0111] In one aspect, the invention provides an aqueous pharmaceutical composition of an anti-PD-1 antibody, comprising: (a) Prorugolimab at a concentration of 100 mg / ml as antibody; (b) trehalose dihydrate at a concentration of 80 mg / ml; (с) L-histidine at a concentration of 0.92 mg / ml; and (d) L-histidine hydrochloride at a concentration of 2.96 mg / ml Including, (e) The above composition relates to an aqueous pharmaceutical composition having a pH of 5.5 to 6.0.

[0073]

[0112] In one aspect, the invention provides an aqueous pharmaceutical composition of an anti-PD-1 antibody, comprising: (a) Prorugolimab at a concentration of 100 mg / ml as antibody; (b) trehalose dihydrate at a concentration of 80 mg / ml; (с) L-histidine at a concentration of 0.92 mg / ml; and (d) L-histidine hydrochloride at a concentration of 2.96 mg / ml Including, (e) The above composition relates to an aqueous pharmaceutical composition having a pH of 5.5.

[0074]

[0113] In one aspect, the invention relates to an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

[0075] [Table 1]

[0076]

[0114] In one aspect, the invention relates to an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

[0077] [Table 2]

[0078]

[0115] The aqueous pharmaceutical composition of the anti-PD-1 antibody prorugolimab of the invention may further comprise a suitable solubilizing agent.

[0116] In some embodiments of the present invention, the solubilizing agent may be poloxamer 188.

[0079]

[0117] In some embodiments of the invention, the poloxamer 188 can be present in an amount greater than 0 mg / ml but less than or equal to 1 mg / ml.

[0118] In some embodiments of the invention, the poloxamer 188 may be present in amounts of 0 mg / ml, 0.1 mg / ml, 0.2 mg / ml, 0.3 mg / ml, 0.4 mg / ml, 0.5 mg / ml, 0.6 mg / ml, 0.7 mg / ml, 0.8 mg / ml, 0.9 mg / ml, 1.0 mg / ml.

[0080]

[0119] In one aspect, the present invention provides a method for producing a method for treating a cancer cell comprising: (a) Prorugolimab at a concentration of 20 mg / ml as antibody; (b) trehalose dihydrate at a concentration of 100 mg / ml; (c) sodium acetate trihydrate in a concentration of 0.2 to 2.5 mg / ml; and (d) acetic acid in an amount to give a pH of 4.5 to 5.5; (e) Poloxamer 188 at a concentration greater than 0 mg / ml but less than or equal to 1 mg / ml The present invention relates to an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising the compound.

[0081]

[0120] In one aspect, the present invention provides a method for producing a method for treating a cancer cell comprising: (a) Prorugolimab at a concentration of 20 mg / ml as antibody; (b) trehalose dihydrate at a concentration of 100 mg / ml; (c) sodium acetate trihydrate in a concentration of 1.7 mg / ml to 1.8 mg / ml; and (d) acetic acid in an amount to give a pH of 5.0; (e) Poloxamer 188 at a concentration greater than 0 mg / ml but less than or equal to 1 mg / ml The present invention relates to an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising the compound.

[0082]

[0121] In one aspect, the present invention provides a method for producing a method for treating a cancer cell comprising: (a) Prorugolimab at a concentration of 20 mg / ml as antibody; (b) trehalose dihydrate at a concentration of 100 mg / ml; (с) sodium acetate trihydrate in a concentration of 1.742 mg / ml; and (d) acetic acid in an amount to give a pH of 5.0; (e) Poloxamer 188 at a concentration greater than 0 mg / ml but less than or equal to 1 mg / ml The present invention relates to an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising the compound.

[0083]

[0122] In one aspect, the present invention provides a method for producing a method for treating a cancer cell comprising: (a) Prorugolimab at a concentration of 100 mg / ml as antibody; (b) trehalose dihydrate at a concentration of 80 mg / ml; (c) sodium acetate trihydrate in a concentration between 0.2 and 2.5 mg / ml; and (d) acetic acid in an amount to give a pH of 4.5 to 5.5; (e) Poloxamer 188 at a concentration greater than 0 mg / ml but less than or equal to 1 mg / ml The present invention relates to an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising the compound.

[0084]

[0123] In one aspect, the present invention provides a method for producing a method for treating a cancer cell comprising: (a) Prorugolimab at a concentration of 100 mg / ml as antibody; (b) trehalose dihydrate at a concentration of 80 mg / ml; (c) sodium acetate trihydrate in a concentration of 1.7 mg / ml to 1.8 mg / ml; and (d) acetic acid in an amount to give a pH of 5.0 to 5.5; (e) Poloxamer 188 at a concentration greater than 0 mg / ml but less than or equal to 1 mg / ml The present invention relates to an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising the compound.

[0085]

[0124] In one aspect, the present invention provides a method for producing a method for treating a cancer cell comprising: (a) Prorugolimab at a concentration of 100 mg / ml as antibody; (b) trehalose dihydrate at a concentration of 80 mg / ml; (с) sodium acetate trihydrate in a concentration of 1.742 mg / ml; and (d) acetic acid in an amount to give a pH of 5.0 to 5.5; (e) Poloxamer 188 at a concentration greater than 0 mg / ml but less than or equal to 1 mg / ml The present invention relates to an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising the compound.

[0086]

[0125] In one aspect, the present invention provides a method for producing a method for treating a cancer cell comprising: (a) Prorugolimab at a concentration of 100 mg / ml as antibody; (b) trehalose dihydrate at a concentration of 80 mg / ml; (с) sodium acetate trihydrate in a concentration of 1.742 mg / ml; and (d) acetic acid in an amount to give a pH of 5.0; (e) Poloxamer 188 at a concentration greater than 0 mg / ml but less than or equal to 1 mg / ml The present invention relates to an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising the compound.

[0087]

[0126] In one aspect, the invention provides an aqueous pharmaceutical composition of an anti-PD-1 antibody, comprising: (a) Prorugolimab at a concentration of 20 mg / ml as antibody; (b) trehalose dihydrate at a concentration of 100 mg / ml; (с) L-histidine in a concentration of 0.2 to 2.5 mg / ml; and (d) L-histidine hydrochloride at a concentration of 0.2 to 3.5 mg / ml Including, (e) the composition has a pH of 5.5 to 6.5; (f) An aqueous pharmaceutical composition comprising poloxamer 188 at a concentration greater than 0 mg / ml but less than or equal to 1 mg / ml.

[0088]

[0127] In one aspect, the invention provides an aqueous pharmaceutical composition of an anti-PD-1 antibody, comprising: (a) Prorugolimab at a concentration of 20 mg / ml as antibody; (b) trehalose dihydrate at a concentration of 100 mg / ml; (с) L-histidine at a concentration of 0.92 mg / ml; and (d) L-histidine hydrochloride at a concentration of 2.96 mg / ml Including, (e) the composition has a pH of 5.5; (f) An aqueous pharmaceutical composition comprising poloxamer 188 at a concentration greater than 0 mg / ml but less than or equal to 1 mg / ml.

[0089]

[0128] In one aspect, the invention provides an aqueous pharmaceutical composition of an anti-PD-1 antibody, comprising: (a) Prorugolimab at a concentration of 100 mg / ml as antibody; (b) trehalose dihydrate at a concentration of 80 mg / ml; (с) L-histidine in a concentration of 0.2 to 2.5 mg / ml; and (d) L-histidine hydrochloride at a concentration of 0.2 to 3.5 mg / ml Including, (e) the composition has a pH of 5.5 to 6.5; (f) An aqueous pharmaceutical composition comprising poloxamer 188 at a concentration greater than 0 mg / ml but less than or equal to 1 mg / ml.

[0090]

[0129] In one aspect, the invention provides an aqueous pharmaceutical composition of an anti-PD-1 antibody, comprising: (a) Prorugolimab at a concentration of 100 mg / ml as antibody; (b) trehalose dihydrate at a concentration of 80 mg / ml; (с) L-histidine at a concentration of 0.92 mg / ml; and (d) L-histidine hydrochloride at a concentration of 2.96 mg / ml Including, (e) the composition has a pH of 5.5 to 6.0; (f) An aqueous pharmaceutical composition comprising poloxamer 188 at a concentration greater than 0 mg / ml but less than or equal to 1 mg / ml.

[0091]

[0130] In one aspect, the invention provides an aqueous pharmaceutical composition of an anti-PD-1 antibody, comprising: (a) Prorugolimab at a concentration of 100 mg / ml as antibody; (b) trehalose dihydrate at a concentration of 80 mg / ml; (с) L-histidine at a concentration of 0.92 mg / ml; and (d) L-histidine hydrochloride at a concentration of 2.96 mg / ml Including, (e) the composition has a pH of 5.5; (f) An aqueous pharmaceutical composition comprising poloxamer 188 at a concentration greater than 0 mg / ml but less than or equal to 1 mg / ml.

[0092]

[0131] In some embodiments of the invention, the poloxamer 188 may be present in amounts of 0 mg / ml, 0.1 mg / ml, 0.2 mg / ml, 0.3 mg / ml, 0.4 mg / ml, 0.5 mg / ml, 0.6 mg / ml, 0.7 mg / ml, 0.8 mg / ml, 0.9 mg / ml, 1.0 mg / ml.

[0093]

[0132] In some aspects, aqueous pharmaceutical compositions of the anti-PD-1 antibodies of the invention may be administered parenterally.

[0133] In some aspects, the aqueous pharmaceutical composition of the anti-PD-1 antibody of the invention may be administered intramuscularly.

[0094]

[0134] In some aspects, the aqueous pharmaceutical composition of the anti-PD-1 antibody of the invention may be administered subcutaneously.

[0135] In some aspects, aqueous pharmaceutical compositions of the anti-PD-1 antibodies of the invention may be administered intravenously.

[0095]

[0136] In some embodiments, the aqueous pharmaceutical composition of the anti-PD-1 antibody of the invention may be administered intravenously as an infusion.

[0137] In some aspects, an aqueous pharmaceutical composition of an anti-PD-1 antibody of the invention may be administered intravenously as an infusion over a 60 minute period; if well tolerated, the infusion time can be shortened to 30 minutes.

[0096]

[0138] In one embodiment, the aqueous pharmaceutical composition of the anti-PD-1 antibody prorugolimab of the invention may be present in a vial.

[0139] In some aspects, the vial may be a glass vial.

[0097]

[0140] In some embodiments, the vial may have a volume of 1 ml to 50 ml.

[0141] In some embodiments, the vial may have a volume of 1 ml to 20 ml.

[0142] In some embodiments, the vial may have a volume of 1 ml, 2 ml, 3 ml, 4 ml, 5 ml, 6 ml, 7 ml, 8 ml, 9 ml, 10 ml, 15 ml, 20 ml, 25 ml, 30 ml, 35 ml, 40 ml, 45 ml or 50 ml.

[0098]

[0143] In one embodiment, the aqueous pharmaceutical composition of the anti-PD-1 antibody prorugolimab of the invention may be present in a syringe.

[0144] In some embodiments, the syringe may have a 1 ml capacity.

[0099]

[0145] In some embodiments, the syringe may have a 2 ml capacity.

[0146] In one aspect, the aqueous pharmaceutical composition of the anti-PD-1 antibody prorugolimab of the invention may be present in a pre-filled syringe.

[0100]

[0147] In some embodiments, the pre-filled syringe may have a 1 ml capacity.

[0148] In some embodiments, the pre-filled syringe may have a capacity of 2 ml.

[0149] Another broad aspect of the invention is a method of producing an aqueous pharmaceutical composition suitable for administration to a subject to inhibit PD-1 protein activity. The method includes combining a pharma- ceutical effective amount of the anti-PD-1 antibody prorugolimab with an acetate-based buffer and an effective amount of trehalose. The method includes combining a pharma- ceutical effective amount of the anti-PD-1 antibody prorugolimab with a histidine-based buffer and an effective amount of trehalose.

[0101]

[0150] In some embodiments, poloxamer 188 can be added as a solubilizing agent.

[0151] Another broad aspect of the invention is the use of an aqueous pharmaceutical composition of the anti-PD-1 antibody prorugolimab, as defined herein, to treat a malignant neoplasm.

[0102]

[0152] In some aspects of the invention, the use of an aqueous pharmaceutical composition of the anti-PD-1 antibody prorugolimab is directed to the treatment of: inoperable or metastatic melanoma, including melanoma at early stage before and after curative treatment; lung cancer, including non-small cell lung cancer, including inoperable or metastatic non-small cell lung cancer (NSCLC); non-squamous non-small cell lung cancer, squamous cell lung cancer; small cell lung cancer, including inoperable or metastatic small cell lung cancer; early stage lung cancer, including before and after curative treatment; metastatic cervical cancer, including early stage cervical cancer, including before and after curative treatment; head and neck tumors, including head and neck squamous cell carcinoma; lymphoma; gastric and intestinal tumors, metastatic squamous cell esophageal cancer; metastatic urothelial carcinoma, bladder cancer including renal cancer; metastatic endometrial cancer, including early stage endometrial cancer before and after curative treatment; metastatic breast cancer, including early stage endometrial cancer before and after curative treatment; metastatic or inoperable liver cancer, including early stage liver cancer before and after curative treatment; inoperable or metastatic solid tumors, including inoperable or metastatic solid tumors with signs of microsatellite instability.

[0103]

[0153] Another broad aspect of the present invention is the use of an aqueous pharmaceutical composition of the anti-PD-1 antibody prorugolimab for the treatment of a malignant neoplasm, comprising administering a pharmacologic effective amount of the aqueous pharmaceutical composition as defined herein.

[0104]

[0154] Another broad aspect of the invention is a method for producing a cellular endothelium comprising: (a) Prorugolimab at a concentration of 15 mg / ml to 40 mg / ml as an antibody; (b) trehalose dihydrate at a concentration of 80 mg / ml to 110 mg / ml; (c) sodium acetate trihydrate in a concentration between 0.2 mg / ml and 2.5 mg / ml; and (d) Acetic acid in an amount that makes the pH 4.5 to 5.5 and (b) administering to a subject in need of treatment an anti-PD-1 antibody comprising an anti-PD-1 antibody, the anti-PD-1 antibody comprising an anti-PD-1 antibody and an anti-PD-1 antibody composition ...

[0105]

[0155] In some aspects of the invention, the prorugolimab may be present at a concentration of 15 mg / ml to 25 mg / ml.

[0156] In some aspects of the invention, the prorugolimab may be present at a concentration of 20 mg / ml.

[0106]

[0157] In some embodiments of the invention, the trehalose dihydrate may be present in a concentration of from 95 mg / ml to 105 mg / ml.

[0158] In some aspects of the invention, the trehalose dihydrate may be present at a concentration of 100 mg / ml.

[0107]

[0159] In some aspects of the invention, the sodium acetate trihydrate may be present in a concentration of 1.6 mg / ml to 1.9 mg / ml.

[0160] In some aspects of the invention, the sodium acetate trihydrate may be present in a concentration of 1.7 mg / ml to 1.8 mg / ml.

[0108]

[0161] In some aspects of the invention, the sodium acetate trihydrate may be present at a concentration of 1.742 mg / ml.

[0162] In some embodiments of the invention, the acetic acid may be added to bring the pH to 5.0.

[0109]

[0163] In some aspects of the invention, the acetic acid may be present in a concentration of 0.04 mg / ml to 0.77 mg / ml.

[0164] In some aspects of the invention, the acetic acid may be present at a concentration of 0.40 mg / ml to 0.50 mg / ml.

[0110]

[0165] In some aspects of the invention, the acetic acid may be present at a concentration of 0.43 mg / ml.

[0166] Another broad aspect of the invention is a method for producing a cellular endothelium comprising: (a) Prorugolimab at a concentration of 5 mg / ml to 40 mg / ml as an antibody; (b) trehalose dihydrate at a concentration of 70 mg / ml to 110 mg / ml; (с) L-histidine in a concentration of 0.2 to 2.5 mg / ml; and (d) L-histidine hydrochloride at a concentration of 0.2 to 3.5 mg / ml and (b) administering to a subject in need of treatment an anti-PD-1 antibody comprising an anti-PD-1 antibody, the anti-PD-1 antibody comprising an anti-PD-1 antibody and an anti-PD-1 antibody composition ...

[0111]

[0167] In some aspects of the invention, the prorugolimab may be present at a concentration of 15 mg / ml to 25 mg / ml.

[0168] In some aspects of the invention, the prorugolimab may be present at a concentration of 20 mg / ml.

[0112]

[0169] In some embodiments of the invention, the trehalose dihydrate may be present in a concentration of from 95 mg / ml to 105 mg / ml.

[0170] In some aspects of the invention, the trehalose dihydrate may be present at a concentration of 100 mg / ml.

[0113]

[0171] In some embodiments of the present invention, the L-histidine may be present at a concentration of 0.7 mg / ml to 1.0 mg / ml.

[0172] In some aspects of the invention, the L-histidine may be present at a concentration of 0.92 mg / ml.

[0114]

[0173] In some aspects of the invention, the L-histidine hydrochloride may be present at a concentration of 2.8 mg / ml to 3.3 mg / ml.

[0174] In some aspects of the invention, the L-histidine hydrochloride may be present at a concentration of 2.96 mg / ml.

[0115]

[0175] In some embodiments of the invention, the composition may have a pH of 5.5 to 6.5.

[0176] In some aspects of the invention, the composition may have a pH of 5.5.

[0116]

[0177] In one aspect, (a) Prorugolimab at a concentration of 20 mg / ml as antibody; (b) trehalose dihydrate at a concentration of 100 mg / ml; (c) sodium acetate trihydrate in a concentration between 0.2 and 2.5 mg / ml; and (d) Acetic acid in an amount that makes the pH 4.5 to 5.5 The present invention provides a use of an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

[0117]

[0178] In one aspect, (a) Prorugolimab at a concentration of 20 mg / ml as antibody; (b) trehalose dihydrate at a concentration of 100 mg / ml; (с) sodium acetate trihydrate in a concentration of 1.7 mg / ml to 1.8 mg / ml; and (d) Acetic acid in an amount that makes the pH 5.0 The present invention provides a use of an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

[0118]

[0179] In one aspect, (a) Prorugolimab at a concentration of 20 mg / ml as antibody; (b) trehalose dihydrate at a concentration of 100 mg / ml; (с) sodium acetate trihydrate in a concentration of 1.742 mg / ml; and (d) Acetic acid in an amount that makes the pH 5.0 The present invention provides a use of an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

[0119]

[0180] In one aspect, there is provided an aqueous pharmaceutical composition of an anti-PD-1 antibody, comprising: (a) Prorugolimab at a concentration of 20 mg / ml as antibody; (b) trehalose dihydrate at a concentration of 100 mg / ml; (с) L-histidine in a concentration of 0.2 to 2.5 mg / ml; and (d) L-histidine hydrochloride at a concentration of 0.2 to 3.5 mg / ml; Including, (e) the composition has a pH of 5.5 to 6.5; Use of the aqueous pharmaceutical composition for treating a malignant neoplasm in a subject in need of such treatment is provided.

[0120]

[0181] In one aspect, there is provided an aqueous pharmaceutical composition of an anti-PD-1 antibody, comprising: (a) Prorugolimab at a concentration of 20 mg / ml as antibody; (b) trehalose dihydrate at a concentration of 100 mg / ml; (с) L-histidine at a concentration of 0.92 mg / ml; and (d) L-histidine hydrochloride at a concentration of 2.96 mg / ml Including, (e) the composition has a pH of 5.5 Use of the aqueous pharmaceutical composition for treating a malignant neoplasm in a subject in need of such treatment is provided.

[0121]

[0182] In one aspect, there is provided a use of an aqueous pharmaceutical composition of an anti-PD-1 antibody for treating a malignant neoplasm in a subject in need of such treatment, comprising:

[0122] [Table 3]

[0123]

[0183] In one aspect, there is provided a use of an aqueous pharmaceutical composition of an anti-PD-1 antibody for treating a malignant neoplasm in a subject in need of such treatment, comprising:

[0124] [Table 4]

[0125]

[0184] The aqueous pharmaceutical composition of the anti-PD-1 antibody prorugolimab of the invention may further comprise a suitable solubilizing agent.

[0185] In some embodiments of the present invention, the solubilizing agent may be poloxamer 188.

[0126]

[0186] In some embodiments of the invention, the poloxamer 188 can be present in an amount greater than 0 mg / ml but less than or equal to 1 mg / ml.

[0187] In some embodiments of the invention, the poloxamer 188 may be present in amounts of 0 mg / ml, 0.1 mg / ml, 0.2 mg / ml, 0.3 mg / ml, 0.4 mg / ml, 0.5 mg / ml, 0.6 mg / ml, 0.7 mg / ml, 0.8 mg / ml, 0.9 mg / ml, 1.0 mg / ml.

[0127]

[0188] In some aspects of the invention, use of the aqueous pharmaceutical composition of the anti-PD-1 antibody prorugolimab may comprise administering the composition at a dose of 1 mg / kg body weight of the anti-PD-1 antibody prorugolimab.

[0128]

[0189] In some aspects of the invention, use of the aqueous pharmaceutical composition of the anti-PD-1 antibody prorugolimab may comprise administering the composition at a dose of 3 mg / kg body weight of the anti-PD-1 antibody prorugolimab.

[0129]

[0190] In some aspects of the invention, use of the aqueous pharmaceutical composition of the anti-PD-1 antibody prorugolimab may comprise administering the composition every two weeks.

[0191] In some aspects of the invention, use of the aqueous pharmaceutical composition of the anti-PD-1 antibody prorugolimab may comprise administering the composition once every two weeks.

[0130]

[0192] In some aspects of the invention, use of the aqueous pharmaceutical composition of the anti-PD-1 antibody prorugolimab may comprise administering the composition every three weeks.

[0193] In some aspects of the invention, use of the aqueous pharmaceutical composition of the anti-PD-1 antibody prorugolimab may comprise administering the composition once every three weeks.

[0131]

[0194] In some aspects of the invention, use of the aqueous pharmaceutical composition of the anti-PD-1 antibody prorugolimab may comprise administering the composition every two weeks at a dose of 1 mg / kg body weight of the anti-PD-1 antibody prorugolimab.

[0132]

[0195] In some aspects of the invention, use of the aqueous pharmaceutical composition of the anti-PD-1 antibody prorugolimab may comprise administering the composition once every two weeks at a dose of 1 mg / kg body weight of the anti-PD-1 antibody prorugolimab.

[0133]

[0196] In some aspects of the invention, use of the aqueous pharmaceutical composition of the anti-PD-1 antibody prorugolimab may comprise administering the composition at a dose of 3 mg / kg body weight of the anti-PD-1 antibody prorugolimab every three weeks.

[0134]

[0197] In some aspects of the invention, use of the aqueous pharmaceutical composition of the anti-PD-1 antibody prorugolimab may comprise administering the composition once every three weeks at a dose of 3 mg / kg body weight of the anti-PD-1 antibody prorugolimab.

[0135]

[0198] In some aspects of the invention, the use of the aqueous pharmaceutical composition of the anti-PD-1 antibody prorugolimab may include administering the composition parenterally.

[0199] In some aspects of the invention, the parenteral administration may be intravenous, subcutaneous, or intramuscular.

[0136]

[0200] In some aspects of the invention, the use of the aqueous pharmaceutical composition of the anti-PD-1 antibody prorugolimab may include administering the composition intravenously as an infusion.

[0201] In some aspects, the aqueous pharmaceutical composition of the anti-PD-1 antibody prorugolimab of the invention can be administered intravenously as an infusion over a 60 minute period; if well tolerated, the infusion time can be shortened to 30 minutes.

[0137]

[0202] In some aspects of the invention, the malignant neoplasm is melanoma, including inoperable or metastatic melanoma, early stage melanoma before and after curative treatment; lung cancer, non-small cell lung cancer, including inoperable or metastatic non-small cell lung cancer (NSCLC).

[0138]

[0203] In one aspect, (a) Prorugolimab at a concentration of 15 mg / ml to 40 mg / ml as an antibody; (b) trehalose dihydrate at a concentration of 80 mg / ml to 110 mg / ml; (c) sodium acetate trihydrate in a concentration between 0.2 mg / ml and 2.5 mg / ml; and (d) Acetic acid in an amount that makes the pH 4.5 to 5.5 In one embodiment, a method is provided for treating a malignant neoplasm in a subject in need of such treatment, comprising administering a therapeutically effective amount of an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

[0139]

[0204] In some aspects of the invention, the prorugolimab may be present at a concentration of 15 mg / ml to 25 mg / ml.

[0205] In some aspects of the invention, the prorugolimab may be present at a concentration of 20 mg / ml.

[0140]

[0206] In some embodiments of the invention, the trehalose dihydrate may be present in a concentration of from 95 mg / ml to 105 mg / ml.

[0207] In some aspects of the invention, the trehalose dihydrate may be present at a concentration of 100 mg / ml.

[0141]

[0208] In some aspects of the invention, the sodium acetate trihydrate may be present in a concentration of 1.6 mg / ml to 1.9 mg / ml.

[0209] In some aspects of the invention, the sodium acetate trihydrate may be present in a concentration of 1.7 mg / ml to 1.8 mg / ml.

[0142]

[0210] In some aspects of the invention, the sodium acetate trihydrate may be present at a concentration of 1.742 mg / ml.

[0211] In some embodiments of the invention, the acetic acid may be added to bring the pH to 5.0.

[0143]

[0212] In some aspects of the invention, the acetic acid may be present in a concentration of 0.04 mg / ml to 0.77 mg / ml.

[0213] In some aspects of the invention, the acetic acid may be present at a concentration of 0.40 mg / ml to 0.50 mg / ml.

[0144]

[0214] In some aspects of the invention, the acetic acid may be present at a concentration of 0.43 mg / ml.

[0215] In one aspect, (a) Prorugolimab at a concentration of 5 mg / ml to 40 mg / ml as an antibody; (b) trehalose dihydrate at a concentration of 70 mg / ml to 110 mg / ml; (с) L-histidine in a concentration of 0.2 to 2.5 mg / ml; and (d) L-histidine hydrochloride at a concentration of 0.2 to 3.5 mg / ml In one embodiment, a method is provided for treating a malignant neoplasm in a subject in need of such treatment, comprising administering a therapeutically effective amount of an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

[0145]

[0216] In some aspects of the invention, the prorugolimab may be present at a concentration of 15 mg / ml to 25 mg / ml.

[0217] In some aspects of the invention, the prorugolimab may be present at a concentration of 20 mg / ml.

[0146]

[0218] In some embodiments of the invention, the trehalose dihydrate may be present in a concentration of from 95 mg / ml to 105 mg / ml.

[0219] In some aspects of the invention, the trehalose dihydrate may be present at a concentration of 100 mg / ml.

[0147]

[0220] In some embodiments of the present invention, the L-histidine may be present at a concentration of 0.7 mg / ml to 1.0 mg / ml.

[0221] In some aspects of the invention, the L-histidine may be present at a concentration of 0.92 mg / ml.

[0148]

[0222] In some aspects of the invention, the L-histidine hydrochloride may be present at a concentration of 2.8 mg / ml to 3.3 mg / ml.

[0223] In some aspects of the invention, the L-histidine hydrochloride may be present at a concentration of 2.96 mg / ml.

[0149]

[0224] In some embodiments of the invention, the composition may have a pH of 5.5 to 6.5.

[0225] In some aspects of the invention, the composition may have a pH of 5.5.

[0150]

[0226] In one aspect, (a) Prorugolimab at a concentration of 20 mg / ml as antibody; (b) trehalose dihydrate at a concentration of 100 mg / ml; (c) sodium acetate trihydrate in a concentration between 0.2 and 2.5 mg / ml; and (d) Acetic acid in an amount that makes the pH 4.5 to 5.5 In one embodiment, a method is provided for treating a malignant neoplasm in a subject in need of such treatment, comprising administering a therapeutically effective amount of an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

[0151]

[0227] In one aspect, (a) Prorugolimab at a concentration of 20 mg / ml as antibody; (b) trehalose dihydrate at a concentration of 100 mg / ml; (с) sodium acetate trihydrate in a concentration of 1.7 mg / ml to 1.8 mg / ml; and (d) Acetic acid in an amount that makes the pH 5.0 In one embodiment, a method is provided for treating a malignant neoplasm in a subject in need of such treatment, comprising administering a therapeutically effective amount of an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

[0152]

[0228] In one aspect, (a) Prorugolimab at a concentration of 20 mg / ml as antibody; (b) trehalose dihydrate at a concentration of 100 mg / ml; (с) sodium acetate trihydrate in a concentration of 1.742 mg / ml; and (d) Acetic acid in an amount that makes the pH 5.0 In one embodiment, a method is provided for treating a malignant neoplasm in a subject in need of such treatment, comprising administering a therapeutically effective amount of an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

[0153]

[0229] In one aspect, there is provided an aqueous pharmaceutical composition of a therapeutically effective amount of an anti-PD-1 antibody, comprising: (a) Prorugolimab at a concentration of 20 mg / ml as antibody; (b) trehalose dihydrate at a concentration of 100 mg / ml; (с) L-histidine in a concentration of 0.2 to 2.5 mg / ml; and (d) L-histidine hydrochloride at a concentration of 0.2 to 3.5 mg / ml Including, (e) the composition has a pH of 5.5 to 6.5; A method for treating a malignant neoplasm in a subject in need thereof comprising administering the aqueous pharmaceutical composition is provided.

[0154]

[0230] In one aspect, there is provided an aqueous pharmaceutical composition of a therapeutically effective amount of an anti-PD-1 antibody, comprising: (a) Prorugolimab at a concentration of 20 mg / ml as antibody; (b) trehalose dihydrate at a concentration of 100 mg / ml; (с) L-histidine at a concentration of 0.92 mg / ml; and (d) L-histidine hydrochloride at a concentration of 2.96 mg / ml Including, (e) the composition has a pH of 5.5 A method for treating a malignant neoplasm in a subject in need thereof comprising administering the aqueous pharmaceutical composition is provided.

[0155]

[0231] In one aspect, a method is provided for treating a malignant neoplasm in a subject in need thereof, comprising administering a therapeutically effective amount of an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

[0156] [Table 5]

[0157]

[0232] In one aspect, a method is provided for treating a malignant neoplasm in a subject in need thereof, comprising administering a therapeutically effective amount of an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

[0158] [Table 6]

[0159]

[0233] The aqueous pharmaceutical composition of the anti-PD-1 antibody prorugolimab of the invention may further comprise a suitable solubilizing agent.

[0234] In some embodiments of the present invention, the solubilizing agent may be poloxamer 188.

[0160]

[0235] In some embodiments of the invention, the poloxamer 188 can be present in an amount greater than 0 mg / ml but less than or equal to 1 mg / ml.

[0236] In some embodiments of the invention, the poloxamer 188 may be present in amounts of 0 mg / ml, 0.1 mg / ml, 0.2 mg / ml, 0.3 mg / ml, 0.4 mg / ml, 0.5 mg / ml, 0.6 mg / ml, 0.7 mg / ml, 0.8 mg / ml, 0.9 mg / ml, 1.0 mg / ml.

[0161]

[0237] In some aspects of the present invention, a method for treating a malignant neoplasm in a subject in need of such treatment may comprise administering a therapeutically effective amount of the above-described aqueous pharmaceutical composition of the anti-PD-1 antibody prorugolimab at a dose of 1 mg / kg body weight of the anti-PD-1 antibody prorugolimab.

[0162]

[0238] In some aspects of the present invention, a method for treating a malignant neoplasm in a subject in need of such treatment may comprise administering a therapeutically effective amount of the above-described aqueous pharmaceutical composition of the anti-PD-1 antibody prorugolimab, at a dose of 3 mg / kg body weight of the anti-PD-1 antibody prorugolimab.

[0163]

[0239] In some aspects of the present invention, a method for treating a malignant neoplasm in a subject in need thereof may comprise administering a therapeutically effective amount of the above-described aqueous pharmaceutical composition of the anti-PD-1 antibody prorugolimab every two weeks.

[0164]

[0240] In some aspects of the present invention, a method for treating a malignant neoplasm in a subject in need thereof may comprise administering a therapeutically effective amount of the above-described aqueous pharmaceutical composition of the anti-PD-1 antibody prorugolimab once every two weeks.

[0165]

[0241] In some aspects of the present invention, a method for treating a malignant neoplasm in a subject in need thereof may comprise administering a therapeutically effective amount of the above-described aqueous pharmaceutical composition of the anti-PD-1 antibody prorugolimab every three weeks.

[0166]

[0242] In some aspects of the present invention, a method for treating a malignant neoplasm in a subject in need thereof may comprise administering a therapeutically effective amount of the above-described aqueous pharmaceutical composition of the anti-PD-1 antibody prorugolimab once every three weeks.

[0167]

[0243] In some aspects of the present invention, a method for treating a malignant neoplasm in a subject in need thereof may comprise administering a therapeutically effective amount of the above-described aqueous pharmaceutical composition of the anti-PD-1 antibody prorugolimab every two weeks at a dose of 1 mg / kg body weight of the anti-PD-1 antibody prorugolimab.

[0168]

[0244] In some aspects of the present invention, a method for treating a malignant neoplasm in a subject in need thereof may comprise administering a therapeutically effective amount of the above-described aqueous pharmaceutical composition of the anti-PD-1 antibody prorugolimab at a dose of 1 mg / kg body weight of the anti-PD-1 antibody prorugolimab once every two weeks.

[0169]

[0245] In some aspects of the present invention, a method for treating a malignant neoplasm in a subject in need thereof may comprise administering a therapeutically effective amount of the above-described aqueous pharmaceutical composition of the anti-PD-1 antibody prorugolimab at a dose of 3 mg / kg body weight of the anti-PD-1 antibody prorugolimab every three weeks.

[0170]

[0246] In some aspects of the present invention, a method for treating a malignant neoplasm in a subject in need of such treatment may comprise administering a therapeutically effective amount of the above-described aqueous pharmaceutical composition of the anti-PD-1 antibody prorugolimab at a dose of 3 mg / kg body weight of the anti-PD-1 antibody prorugolimab once every three weeks.

[0171]

[0247] In some aspects of the present invention, a method for treating a malignant neoplasm in a subject in need thereof may comprise parenterally administering a therapeutically effective amount of the above-described aqueous pharmaceutical composition of the anti-PD-1 antibody prorugolimab.

[0172]

[0248] In some aspects of the invention, the parenteral administration may be intravenous, subcutaneous, or intramuscular.

[0249] In some aspects of the present invention, a method for treating a malignant neoplasm in a subject in need thereof may comprise administering a therapeutically effective amount of the above-described aqueous pharmaceutical composition of the anti-PD-1 antibody prorugolimab intravenously as an infusion.

[0173]

[0250] In some aspects, the aqueous pharmaceutical composition of the anti-PD-1 antibody prorugolimab of the invention can be administered intravenously as an infusion over a 60 minute period; if well tolerated, the infusion time can be shortened to 30 minutes.

[0174]

[0251] In some aspects of the invention, the malignant neoplasm is melanoma, including inoperable or metastatic melanoma, early stage melanoma before and after curative treatment; lung cancer, non-small cell lung cancer, including inoperable or metastatic non-small cell lung cancer (NSCLC).

[0175]

[0252] In one aspect, a method is provided for treating a malignant neoplasm in a subject in need thereof, comprising administering a therapeutically effective amount of prorugolimab.

[0253] In some aspects of the invention, a method for treating a malignant neoplasm in a subject in need thereof may comprise administering a therapeutically effective amount of prorugolimab at a dose of 1 mg / kg.

[0176]

[0254] In some aspects of the invention, a method for treating a malignant neoplasm in a subject in need thereof may comprise administering a therapeutically effective amount of prorugolimab at a dose of 3 mg / kg.

[0177]

[0255] In some aspects of the invention, a method for treating a malignant neoplasm in a subject in need thereof may comprise administering a therapeutically effective amount of prorugolimab every two weeks.

[0178]

[0256] In some aspects of the invention, a method for treating a malignant neoplasm in a subject in need thereof may comprise administering a therapeutically effective amount of prorugolimab once every two weeks.

[0179]

[0257] In some aspects of the invention, a method for treating a malignant neoplasm in a subject in need thereof may comprise administering a therapeutically effective amount of prorugolimab every three weeks.

[0180]

[0258] In some aspects of the invention, a method for treating a malignant neoplasm in a subject in need thereof may comprise administering a therapeutically effective amount of prorugolimab once every three weeks.

[0181]

[0259] In some aspects of the invention, a method for treating a malignant neoplasm in a subject in need thereof may comprise administering a therapeutically effective amount of prorugolimab at a dose of 1 mg / kg every two weeks.

[0182]

[0260] In some aspects of the invention, a method for treating a malignant neoplasm in a subject in need thereof may comprise administering a therapeutically effective amount of prorugolimab at a dose of 1 mg / kg once every two weeks.

[0183]

[0261] In some aspects of the invention, a method for treating a malignant neoplasm in a subject in need thereof may comprise administering a therapeutically effective amount of prorugolimab at a dose of 3 mg / kg every three weeks.

[0184]

[0262] In some aspects of the invention, a method for treating a malignant neoplasm in a subject in need thereof may comprise administering a therapeutically effective amount of prorugolimab at a dose of 3 mg / kg once every three weeks.

[0185]

[0263] In some aspects of the invention, a method for treating a malignant neoplasm in a subject in need thereof may comprise parenterally administering a therapeutically effective amount of prorugolimab.

[0186]

[0264] In some aspects of the invention, the parenteral administration may be intravenous, subcutaneous, or intramuscular.

[0265] In some aspects of the invention, a method for treating a malignant neoplasm in a subject in need thereof may comprise administering a therapeutically effective amount of prorugolimab intravenously as an infusion.

[0187]

[0266] In some embodiments, prorugolimab may be administered intravenously as an infusion over 60 minutes; if well tolerated, the infusion time can be shortened to 30 minutes.

[0267] In some aspects of the invention, the malignant neoplasm is melanoma, including inoperable or metastatic melanoma, early stage melanoma before and after curative treatment; lung cancer, non-small cell lung cancer, including inoperable or metastatic non-small cell lung cancer (NSCLC).

[0188]

[0268] In one aspect of the invention, there is provided a pharmaceutical composition suitable for administration to a subject for inhibiting PD-1 protein activity, the pharmaceutical composition comprising, for each 1 mL of the pharmaceutical composition:

[0189] [Table 7]

[0190]

[0269] In one aspect of the invention, there is provided a pharmaceutical composition suitable for administration to a subject for inhibiting PD-1 protein activity, the pharmaceutical composition comprising, for each 1 mL of the pharmaceutical composition:

[0191] [Table 8]

[0192]

[0270] In some aspects of the invention, the aqueous pharmaceutical composition suitable for administration to a subject to inhibit PD-1 protein activity may be administered at a dose of 1 mg / kg body weight of the anti-PD-1 antibody prorugolimab.

[0193]

[0271] In some aspects of the invention, the aqueous pharmaceutical composition suitable for administration to a subject to inhibit PD-1 protein activity may be administered at a dose of 3 mg / kg body weight of the anti-PD-1 antibody prorugolimab.

[0194]

[0272] In some aspects of the present invention, the aqueous pharmaceutical composition suitable for administration to a subject to inhibit PD-1 protein activity may be administered every two weeks.

[0273] In some aspects of the present invention, the aqueous pharmaceutical composition suitable for administration to a subject to inhibit PD-1 protein activity may be administered once every two weeks.

[0195]

[0274] In some aspects of the present invention, the aqueous pharmaceutical composition suitable for administration to a subject to inhibit PD-1 protein activity may be administered once every three weeks.

[0275] In some aspects of the invention, the aqueous pharmaceutical composition suitable for administration to a subject to inhibit PD-1 protein activity may be administered at a dose of 1 mg / kg body weight of the anti-PD-1 antibody prorugolimab every two weeks.

[0196]

[0276] In some aspects of the invention, the aqueous pharmaceutical composition suitable for administration to a subject to inhibit PD-1 protein activity may be administered at a dose of 1 mg / kg body weight of the anti-PD-1 antibody prorugolimab once every two weeks.

[0197]

[0277] In some aspects of the invention, the aqueous pharmaceutical composition suitable for administration to a subject to inhibit PD-1 protein activity may be administered at a dose of 3 mg / kg body weight of the anti-PD-1 antibody prorugolimab every three weeks.

[0198]

[0278] In some aspects of the invention, the aqueous pharmaceutical composition suitable for administration to a subject to inhibit PD-1 protein activity may be administered at a dose of 3 mg / kg body weight of the anti-PD-1 antibody prorugolimab once every three weeks.

[0199]

[0279] In some aspects of the invention, a method for treating a malignant neoplasm in a subject in need thereof may comprise parenterally administering a therapeutically effective amount of prorugolimab.

[0200]

[0280] In some aspects of the invention, the parenteral administration may be intravenous, subcutaneous, or intramuscular.

[0281] In some aspects of the invention, a method for treating a malignant neoplasm in a subject in need thereof may comprise administering a therapeutically effective amount of prorugolimab intravenously as an infusion.

[0201]

[0282] In some embodiments, prorugolimab may be administered intravenously as an infusion over 60 minutes; if well tolerated, the infusion time can be shortened to 30 minutes.

[0283] In some aspects of the invention, the malignant neoplasm is melanoma, including inoperable or metastatic melanoma, early stage melanoma before and after curative treatment; lung cancer, non-small cell lung cancer, including inoperable or metastatic non-small cell lung cancer (NSCLC). Exemplary embodiments:

[0284] The present invention relates to a suitable aqueous pharmaceutical composition of the anti-PD-1 antibody prorugolimab. In one embodiment of the present invention, the aqueous pharmaceutical composition of prorugolimab may include an acetate-based buffer and trehalose. Poloxamer 188 may be added as a solubilizing agent. In another embodiment of the present invention, the aqueous pharmaceutical composition of prorugolimab may include a histidine-based buffer and trehalose. Poloxamer 188 may be added as a solubilizing agent.

[0202]

[0285] The histidine-based buffer may be the result of combining L-histidine with histidine hydrochloride, or additionally with hydrochloric acid, or other acids. Although histidine hydrochloride may be used as the salt of the histidine-based buffer, it is understood that any other histidine-based salt may be used for the histidine-based buffer without departing from the present teachings.

[0203]

[0286] The acetate-based buffer may be the result of combining acetic acid with sodium acetate trihydrate. Although sodium acetate trihydrate may be used as the salt of the acetate-based buffer, it is understood that any other acetate salt, such as potassium acetate, may be used in the acetate-based buffer without departing from the present teachings.

[0204]

[0287] In addition, the compositions of the present invention may further comprise one or more other suitable excipients well known to those skilled in the art.

[0288] In some aspects, the liquid pharmaceutical composition embodiment is stable during storage in the sense that no further process of protein aggregation or modification thereof occurs relative to the indicator of stability at time zero.

[0205]

[0289] In one embodiment, the present inventors have surprisingly obtained an aqueous pharmaceutical composition having a high concentration of the anti-PD-1 antibody prorugolimab, wherein the prorugolimab can be present in a concentration of 90 mg / ml to 150 mg / ml. In some embodiments of the present invention, the prorugolimab can be present in a concentration of 90 mg / ml, 95 mg / ml, 100 mg / ml, 105 mg / ml, 110 mg / ml, 115 mg / ml, 120 mg / ml, 125 mg / ml, 130 mg / ml, 135 mg / ml, 140 mg / ml, 145 mg / ml, or 150 mg / ml.

[0206]

[0290] Such highly concentrated aqueous pharmaceutical compositions of the anti-PD-1 antibody prorugolimab of the present invention exhibited colloidal stability under vigorous stirring (800 rpm) for 120 hours, high thermal stability under heating at 50° C. and 37° C., and a viscosity of less than 50 cP suitable for parenteral administration.

[0207]

[0291] The compositions are suitable for parenteral administration, such as intravenous, subcutaneous, intradermal, intraarterial, intrathecal, intraperitoneal, intraarticular and / or intramuscular administration.

[0292] The provided pharmaceutical compositions can be administered to an individual in need of treatment by systemic injection, e.g., intravenous or subcutaneous or intramuscular injection, or by injection or application at a suitable site, e.g., if a site is available for surgery, by direct injection or application at the site, or by topical use.

[0208]

[0293] The composition may be administered intravenously as an infusion to a subject in need thereof.

[0294] In some aspects, an aqueous pharmaceutical composition of the anti-PD-1 antibody prorugolimab of the invention may be administered intravenously as an infusion over a 60 minute period; if well tolerated, the infusion time can be shortened to 30 minutes.

[0209]

[0295] After dilution, the aqueous pharmaceutical composition of the anti-PD-1 antibody prorugolimab can be used. For this purpose, the required amount of the composition is transferred from the vial to an injection container containing a sterile 0.9% sodium chloride solution or a sterile 5% glucose solution. The concentration of the above composition in the resulting solution can range from 0.5 mg / ml to 10 mg / ml. The resulting solution is stirred by gently rotating the injection container to avoid foaming. Treatment method and use of the aqueous composition

[0296] In another aspect, the invention relates to a method for treating a mammal, comprising the step of administering to the mammal a therapeutically effective amount of the pharmaceutical composition of the invention, wherein the concentration of the anti-PD1 antibody prorugolimab is from 15 mg / ml to 40 mg / ml, and the mammal may have a disease or disorder that can be effectively treated using the anti-PD-1 antibody prorugolimab of the invention.

[0210]

[0297] In another aspect, the invention relates to a method for treating a mammal, comprising the step of administering to the mammal a therapeutically effective amount of the pharmaceutical composition of the invention, wherein the concentration of the anti-PD-1 antibody prorugolimab is 20 mg / ml, and the mammal may have a disease or disorder that can be effectively treated with the anti-PD-1 antibody prorugolimab of the invention.

[0211]

[0298] In a preferred embodiment, the mammal is a human.

[0299] In another aspect, the present invention relates to a method for treating a subject in need of treatment, comprising the step of administering to the subject a therapeutically effective amount of the pharmaceutical composition of the present invention, wherein the concentration of the anti-PD-1 antibody prorugolimab is from 15 mg / ml to 40 mg / ml, and the subject may have a disease or disorder that can be effectively treated using the anti-PD-1 antibody prorugolimab of the present invention.

[0212]

[0300] In another aspect, the present invention relates to a method for treating a subject in need of treatment, comprising the step of administering to the subject a therapeutically effective amount of the pharmaceutical composition of the present invention, wherein the concentration of the anti-PD-1 antibody prorugolimab is 20 mg / ml, and the subject may have a disease or disorder that can be effectively treated with the anti-PD-1 antibody prorugolimab of the present invention.

[0213]

[0301] In a preferred embodiment, the subject is a human.

[0302] In one aspect, the invention relates to a method for treating melanoma comprising administering to a subject in need thereof a therapeutically effective amount of one of the provided compositions of the invention, wherein the concentration of the anti-PD-1 antibody prorugolimab is between 15 mg / ml and 40 mg / ml.

[0214]

[0303] In one aspect, the invention relates to a method for treating melanoma, comprising administering to a subject in need thereof a therapeutically effective amount of one of the provided compositions of the invention, wherein the concentration of the anti-PD-1 antibody prorugolimab is 20 mg / ml.

[0215]

[0304] In one aspect, the invention relates to a method for treating inoperable melanoma comprising administering to a subject in need thereof a therapeutically effective amount of one of the provided compositions of the anti-PD-1 antibody prorugolimab of the invention, wherein the concentration of the anti-PD-1 antibody prorugolimab is between 15 mg / ml and 40 mg / ml.

[0216]

[0305] In one aspect, the present invention relates to a method for treating inoperable melanoma comprising administering to a subject in need thereof a therapeutically effective amount of one of the provided compositions of the anti-PD-1 antibody prorugolimab of the present invention, wherein the concentration of the anti-PD-1 antibody prorugolimab is 20 mg / ml.

[0217]

[0306] In one aspect, the invention relates to a method for treating metastatic melanoma comprising administering to a subject in need thereof a therapeutically effective amount of one of the provided compositions of the anti-PD-1 antibody prorugolimab of the invention, wherein the concentration of the anti-PD-1 antibody prorugolimab is between 15 mg / ml and 40 mg / ml.

[0218]

[0307] In one aspect, the present invention relates to a method for treating metastatic melanoma comprising administering to a subject in need thereof a therapeutically effective amount of one of the provided compositions of the anti-PD-1 antibody prorugolimab of the present invention, wherein the concentration of the anti-PD-1 antibody prorugolimab is 20 mg / ml.

[0219]

[0308] In one aspect, the invention relates to a method for treating early stage melanoma before or after curative treatment, comprising administering to a subject in need thereof a therapeutically effective amount of one of the provided compositions of the anti-PD-1 antibody prorugolimab of the invention, wherein the concentration of the anti-PD-1 antibody prorugolimab is between 15 mg / ml and 40 mg / ml.

[0220]

[0309] In one aspect, the present invention relates to a method for treating early stage melanoma before or after definitive treatment, comprising administering to a subject in need thereof a therapeutically effective amount of one of the provided compositions of the anti-PD-1 antibody prorugolimab of the present invention, wherein the concentration of the anti-PD-1 antibody prorugolimab is 20 mg / ml.

[0221]

[0310] In one aspect, the invention relates to a method for treating lung cancer comprising administering to a subject in need thereof a therapeutically effective amount of one of the provided compositions of the anti-PD-1 antibody prorugolimab of the invention, wherein the concentration of the anti-PD-1 antibody prorugolimab is between 15 mg / ml and 40 mg / ml.

[0222]

[0311] In one aspect, the invention relates to a method for treating lung cancer comprising administering to a subject in need thereof a therapeutically effective amount of one of the provided compositions of the anti-PD-1 antibody prorugolimab of the invention, wherein the concentration of the anti-PD-1 antibody prorugolimab is 20 mg / ml.

[0223]

[0312] In one aspect, the invention relates to a method for treating non-small cell lung cancer (NSCLC) comprising administering to a subject in need thereof a therapeutically effective amount of one of the provided compositions of the anti-PD-1 antibody prorugolimab of the invention, wherein the concentration of the anti-PD-1 antibody prorugolimab is between 15 mg / ml and 40 mg / ml.

[0224]

[0313] In one aspect, the invention relates to a method for treating non-small cell lung cancer (NSCLC) comprising administering to a subject in need thereof a therapeutically effective amount of one of the provided compositions of the anti-PD-1 antibody prorugolimab of the invention, wherein the concentration of the anti-PD-1 antibody prorugolimab is 20 mg / ml.

[0225]

[0314] In one aspect, the invention relates to a method for treating inoperable or metastatic non-small cell lung cancer comprising administering to a subject in need thereof a therapeutically effective amount of one of the provided compositions of the anti-PD-1 antibody prorugolimab of the invention, wherein the concentration of the anti-PD-1 antibody prorugolimab is between 15 mg / ml and 40 mg / ml.

[0226]

[0315] In one aspect, the present invention relates to a method for treating inoperable or metastatic non-small cell lung cancer, comprising administering to a subject in need thereof a therapeutically effective amount of one of the provided compositions of the anti-PD-1 antibody prorugolimab of the present invention, wherein the concentration of the anti-PD-1 antibody prorugolimab is 20 mg / ml.

[0227]

[0316] In one aspect, a method is provided for treating melanoma in a subject in need thereof, comprising administering a therapeutically effective amount of an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

[0228] [Table 9]

[0229]

[0317] In one aspect, a method is provided for treating melanoma in a subject in need thereof, comprising administering a therapeutically effective amount of an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

[0230] [Table 10]

[0231]

[0318] In one aspect, a method is provided for treating inoperable melanoma in a subject in need thereof, comprising administering a therapeutically effective amount of an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

[0232] [Table 11]

[0233]

[0319] In one aspect, a method is provided for treating inoperable melanoma in a subject in need thereof, comprising administering a therapeutically effective amount of an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

[0234] [Table 12]

[0235]

[0320] In one aspect, a method is provided for treating metastatic melanoma in a subject in need thereof, comprising administering a therapeutically effective amount of an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

[0236] [Table 13]

[0237]

[0321] In one aspect, a method is provided for treating metastatic melanoma in a subject in need thereof, comprising administering a therapeutically effective amount of an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

[0238] [Table 14]

[0239]

[0322] In one aspect, there is provided a method for treating early stage melanoma, pre- or post-definitive treatment, in a subject in need thereof, comprising administering a therapeutically effective amount of an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

[0240] [Table 15]

[0241]

[0323] In one aspect, there is provided a method for treating early stage melanoma, pre- or post-definitive treatment, in a subject in need thereof, comprising administering a therapeutically effective amount of an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

[0242] [Table 16]

[0243]

[0324] In one aspect, a method is provided for treating lung cancer in a subject in need thereof, comprising administering a therapeutically effective amount of an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

[0244] [Table 17]

[0245]

[0325] In one aspect, a method is provided for treating lung cancer in a subject in need thereof, comprising administering a therapeutically effective amount of an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

[0246] [Table 18]

[0247]

[0326] In one aspect, a method is provided for treating non-small cell lung cancer (NSCLC) in a subject in need of such treatment, comprising administering a therapeutically effective amount of an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

[0248] [Table 19]

[0249]

[0327] In one aspect, a method is provided for treating non-small cell lung cancer (NSCLC) in a subject in need of such treatment, comprising administering a therapeutically effective amount of an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

[0250] [Table 20]

[0251]

[0328] In one aspect, a method is provided for treating inoperable or metastatic non-small cell lung cancer in a subject in need thereof, comprising administering a therapeutically effective amount of an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

[0252] [Table 21]

[0253]

[0329] In one aspect, a method is provided for treating inoperable or metastatic non-small cell lung cancer in a subject in need thereof, comprising administering a therapeutically effective amount of an aqueous pharmaceutical composition of an anti-PD-1 antibody comprising:

[0254] [Table 22]

[0255]

[0330] In one aspect, the invention relates to a method for treating melanoma comprising administering a therapeutically effective amount of the anti-PD-1 antibody prorugolimab to a subject in need of such treatment.

[0256]

[0331] In one aspect, the invention relates to a method for treating inoperable melanoma comprising administering a therapeutically effective amount of the anti-PD-1 antibody prorugolimab to a subject in need of such treatment.

[0257]

[0332] In one aspect, the invention relates to a method for treating metastatic melanoma comprising administering a therapeutically effective amount of the anti-PD-1 antibody prorugolimab to a subject in need of such treatment.

[0258]

[0333] In one aspect, the invention relates to a method for treating early stage melanoma before or after definitive treatment, comprising administering a therapeutically effective amount of the anti-PD-1 antibody prorugolimab to a subject in need of such treatment.

[0259]

[0334] In one aspect, the invention relates to a method for treating lung cancer comprising administering a therapeutically effective amount of the anti-PD-1 antibody prorugolimab to a subject in need of such treatment.

[0260]

[0335] In one aspect, the invention relates to a method for treating non-small cell lung cancer (NSCLC), comprising administering a therapeutically effective amount of the anti-PD-1 antibody prorugolimab to a subject in need of such treatment.

[0261]

[0336] In one aspect, the invention relates to a method for treating inoperable or metastatic non-small cell lung cancer, comprising administering a therapeutically effective amount of the anti-PD-1 antibody prorugolimab to a subject in need of such treatment.

[0262]

[0337] The therapeutically effective amount of the anti-PD-1 antibodies of the present invention and aqueous compositions comprising the anti-PD-1 antibody prorugolimab of the present invention in the formulations provided will depend on the condition being treated, the severity of the condition, previous treatments, and the patient's medical history and response to the therapeutic agent. The appropriate dose can be adjusted as determined by the attending physician so that the patient can be administered through one or multiple injections.

[0263]

[0338] In one embodiment, the effective amount of anti-PD-1 antibody per dose for a patient is about 0.01-10 mg / kg body weight, or about 1-10 mg / kg body weight, or about 0.05 mg / kg body weight, or about 0.25 mg / kg body weight, or about 0.5 mg / kg body weight, or about 1 mg / kg body weight, or about 2 mg / kg body weight, or about 3 mg / kg body weight, or about 4 mg / kg body weight, or about 5 mg / kg body weight, or about 6 mg / kg body weight, or about 7 mg / kg body weight, or about 8 mg / kg body weight, or about 9 mg / kg body weight, or about 10 mg / kg body weight.

[0264]

[0339] The frequency of administration can typically be about once a week, or about once every two weeks, or about once every three weeks.

[0340] In another embodiment, acceptable doses for administration by injection may include 5-450 mg / dose, or may include 40 mg, or 50 mg, or 60 mg / dose; or may include 70 mg, or 80 mg, or 90 mg, or 100 mg / dose; or may include 110 mg, or 120 mg, or 130 mg, or 140 mg / dose; or may include 150 mg, or 160 mg, or 170 mg, or 180 mg / dose; or may include 190 mg, or 200 mg, or 210 mg, or 220 mg / dose. or may comprise 230mg, or 240mg, or 250mg, or 260mg per dose; or may comprise 270mg, or 280mg, or 290mg per dose; or may comprise 300mg, or 310mg, or 320mg, or 330mg, or 340mg, or 350mg per dose; or may comprise 360mg, or 370mg, or 380mg, or 390mg, or 400mg, or 410mg, or 420mg, or 430mg, or 440mg, or 450mg per dose.

[0265]

[0341] In one embodiment of the present invention, the dose can be delivered as one or more injections. The dose can be delivered as one, two or three injections. In some embodiments of the present invention, the duration of treatment can be one to several injections. In some embodiments of the present invention, the improvement of the patient's condition can be achieved through long-term treatment. In some embodiments of the present invention, the duration of treatment can be until disease progression or lifelong.

[0266]

[0342] In another aspect, the pharmaceutical compositions of the present invention can be prepared as non-packaged formulations, essentially where the components of the pharmaceutical composition are present in amounts higher than may be required for administration and diluted accordingly prior to administration.

[0267]

[0343] Alternatively, the pharmaceutical compositions can be frozen, spray dried or lyophilized and reconstituted prior to application to a suitable sterile carrier. Lyophilization can be carried out using techniques known in the art, including various steps such as freezing, annealing, primary and secondary drying, etc.

[0268]

[0344] The pharmaceutical composition can be administered as a single therapeutic agent or in combination with additional therapeutic agents as required.Thus, in one aspect, the provided method for treatment and / or prevention is used in combination with the administration of a therapeutically effective amount of another active agent.The other active agent can be administered before, during, or after the administration of the pharmaceutical composition of the present invention.The other active agent can be administered as part of the provided composition or, alternatively, as a separate formulation.

[0269]

[0345] Administration of the provided pharmaceutical compositions can be by a variety of means, including parenteral, oral, buccal, nasal, rectal and topical administration. Parenteral administration can include, but is not limited to, percutaneous, subcutaneous, intravenous, intraarterial, intraperitoneal, intradermal, intracardiac, intraventricular, intracranial, intratracheal, intrathecal administration, intramuscular injection, and intravitreal injection.

[0270]

[0346] The pharmaceutical composition of the present invention is particularly suitable for parenteral administration, i.e. subcutaneous, intramuscular, intravenous, intraperitoneal, intraspinal, intraarticular, intrasynovial and / or intrathecal administration. Parenteral administration can be by bolus injection or continuous infusion. Pharmaceutical compositions for injection can be in standard dosage forms, such as, but not limited to, ampoules, vials, pre-filled syringes, or multi-dose containers with added preservatives. In addition, several recent approaches to drug delivery have been developed, and the pharmaceutical composition of the present invention is suitable for administration by these new methods, including, for example, pen injectors such as BD Physioject™, Inject-ease®, Genject®, GenPen®, and needleless devices such as MediJector® and BioJector®. The pharmaceutical composition of the present invention can also be applied to administration methods that have not yet been opened.

[0271]

[0347] See also Langer, 1990, Science 249:1527-1533.

[0348] The provided pharmaceutical composition can also be formulated as a depot preparation. Such long-acting preparations can be administered by implantation (e.g., subcutaneous or intramuscular) or by intramuscular injection. Thus, for example, the composition can be modified with suitable polymers or hydrophobic materials (e.g., as an emulsion in acceptable oil), or ion exchange resins, or moderately soluble derivatives, e.g., moderately soluble salts.

[0272]

[0349] If desired, pharmaceutical composition can be provided in a vial, package, or dispenser device that can contain one or more unit dosage forms containing active ingredient.In one embodiment, dispenser device can include a syringe that contains a single dose of liquid composition that is ready for injection.Syringe can be accompanied by instructions on administration.

[0273]

[0350] In another aspect, the present invention relates to a kit or container containing the aqueous pharmaceutical composition according to the present invention. The concentration of the antibody in the aqueous pharmaceutical composition may vary over a wide range, but is generally within the range of about 1 to about 200 mg / ml. In addition, the kit may include an instruction manual.

[0274]

[0351] The method of obtaining the composition includes adding to the aqueous phase acetate buffer, followed by adding, in any order, the following components: trehalose, prorugolimab and / or a solubilizer selected from the group of polysorbate 20, polysorbate 80, poloxamer 188 or combinations thereof.

[0275]

[0352] The method of obtaining the composition includes adding to the aqueous phase histidine buffer, followed by adding, in any order, the following components: trehalose, prorugolimab and / or a solubilizing agent selected from the group of polysorbate 20, polysorbate 80, poloxamer 188 or combinations thereof. Illustrative studies:

[0353] The following represents an exemplary study to determine reagents and their concentrations for making an aqueous pharmaceutical composition of the anti-PD-1 antibody prorugolimab of the invention.

[0276]

[0354] The examples and studies presented herein are for illustrative purposes and demonstrate the suitability of certain components used in aqueous pharmaceutical compositions of the anti-PD-1 antibody prorugolimab. It is understood that other methods and techniques may be used by those skilled in the art without departing from the concept of the present invention.

[0277]

[0355] The suitability of the aqueous compositions of the present invention was tested using the exemplary methods described herein.

[0356] Example 1 Preparation of stable anti-PD-1 antibody prorugolimab formulation

[0357] Antibody samples (5 mg / ml) were prepared under pressure in a Stirred Cell (Millipore). For this purpose, the initial antibody formulation was placed in the cell, the protein was concentrated to 10 mg / ml under continuous stirring under a compressed air stream, and then at least 10 times the volume of an aqueous solution containing the target formulation, including buffers, osmotic agents, and additional water-soluble stabilizers if necessary, was added to the cell. After diafiltration, the antibody was concentrated to approximately 10 mg / ml, removed from the cell, and the exact protein concentration was determined by UV spectroscopy. An appropriate solution of excipients was then added to the sample to prepare a solution containing the protein at the target concentration of 5 ± 0.2 mg / ml.

[0278]

[0358] Protein samples with concentrations above 50 mg / ml were prepared in Pellicon cassettes (Millipore) in tangential flow mode. For this purpose, the initial antibody formulation was placed in a diafiltration tank, the protein was concentrated to 50-100 mg / ml, and the system was fed with at least 10 times the volume of a solution containing the target formulation, including buffer, osmotic agent, and, if necessary, additional water-soluble stabilizers. Alternatively, concentrates of osmotic agents and water-soluble stabilizers can be added after diafiltration. After diafiltration, the antibody was concentrated to a concentration above the target concentration, removed from the system, and the exact protein concentration was measured. Then, appropriate solutions of excipients were added to the sample to prepare a solution containing the protein at the target concentration.

[0279]

[0359] When obtaining a formulation containing a solubilizer, a detergent concentrate was added to the antibody after diafiltration and concentration with a solution of excipient along with the final dilution of the antibody to the target concentration.

[0360] The antibody solution was filtered using a 0.22 μm membrane before being aseptically filled into the final container (eg, glass / plastic container, vial or syringe).

[0280]

[0361] Example 2 Protein concentration assay in test samples

[0362] Protein concentrations were determined by UV spectroscopy at a wavelength of 280 nm in UV-clear plates.

[0281]

[0363] Each sample was diluted to approximately 0.5 mg / ml with an appropriate solution of excipient. 150 μl of the diluted sample was placed in a UV spectrophotometer plate well. The optical density of the solution in the plate well was measured using a plate spectrophotometer at a wavelength of 280 nm. The appropriate solution of excipient was used as the reference solution.

[0282]

[0364] The protein concentration (C) (mg / ml) was calculated by the following formula:

[0283]

number

[0284] During the ceremony, A 280 is the value of optical density at a wavelength of 280 nm; ε is the extinction coefficient of the test protein; b is the total dilution factor of the sample; l is the layer thickness in the plate well, for 150 μl, l=0.42 cm.

[0285]

[0365] Example 3 PEG Aggregation Studies

[0366] A solution of 20-25% by mass of PEG 6000 in the test excipient composition was prepared. The resulting solution was filtered through a Durapore 0.45 μm filter.

[0286]

[0367] Estimated volumes of samples, excipient solutions, and 20-25% PEG6000 solution were transferred to a 96-well UV plate, such that the concentration of PEG6000 in some wells ranged from 0-18% and the protein concentration in each well was 1 mg / ml. All the resulting solutions in the wells were mixed thoroughly by pipetting.

[0287]

[0368] The turbidity of the solutions was then visually assessed and the optical density of the solutions was measured at a wavelength of 400 nm.

[0369] Protein precipitation in the presence of PEG is associated with the effect of volume displacement, i.e. the protein is sterically excluded from the solvent domain by the polymer. This increases the protein concentration until it exceeds its solubility and precipitates. The less stable the sample, the lower the PEG6000 concentration at which the sample forms visible aggregates (emulsions).

[0288]

[0370] Example 4 Evaluation of colloidal stability by shaking test

[0371] The test samples were divided into two aliquots of 200 μl and placed into glass vials, one vial per formulation was transferred to a refrigerator for degradation at 2-8°C, and the remaining vials were placed into a shaker and shaken at 800 rpm for the prescribed period at 2-8°C. After stress, the vials were vortexed and transferred for analysis.

[0289]

[0372] Example 5 Assessment of colloidal stability by cryoconcentration

[0373] The test samples were divided into two portions and placed in plastic vials: one vial per formulation was stored in a refrigerator at 2-8°C, and the remaining vial was stored for the indicated time in a freezer at -16-20°C. After stress, the vials were removed from the freezer and kept at room temperature until the contents were completely thawed, then the solutions were vortexed and transferred for analysis.

[0290]

[0374] Example 6 Evaluation of thermal stability by heat stress

[0375] The test samples were divided into two portions and placed in separate glass vials: one vial per composition was kept in a refrigerator at 2-8°C, while the remaining vial was incubated in a thermostat at the required temperature for the prescribed time. After heating, the vials were removed from the thermostat, kept at room temperature for approximately 15 minutes, and transferred for analysis.

[0291]

[0376] Example 7 Determination of sample homogeneity by size-exclusion high performance liquid chromatography (SEC HPLC) Tosoh column TSK-GelG3000SW XL 7.8mm ID x 30cm, Catalog No. 08541 Column temperature: 25℃ Mobile phase flow rate: 0.7ml / min Injection volume: 10μl Sample concentration: 5mg / ml Detector wavelength: 220nm Elution time: 25 min

[0292] [Table 23]

[0293] The mobile phase pH was adjusted to 7.0 with orthophosphoric acid. The change in purity after stress was calculated by the following formula: Δ=(destruction of main peak after stress-destruction of main peak before stress)

[0377] Example 8 Determination of sample charge heterogeneity (charge form profile) using Labchip GX II and Caliper

[0378] Preparation of test samples.

[0294]

[0379] The samples were diluted to a concentration of 1 mg / ml. 2 μl of 5 mg / ml Carboxypeptidase B (CpB) solution was added to 200 μl of the resulting solution, and the solution was mixed and incubated at 37° C. for 2 hours. The test samples were dialyzed against three changes of water. For dialysis, the test solutions were placed in 0.5 ml Amicon ultracentrifuge tubes and centrifuged for 10 minutes at 10,000 rpm in an Eppendorf Centrifuge 5417R. The intensity of the solution absorption against water was measured using a Cary 50 Bio spectrophotometer. A test series of probes was prepared at a concentration of 2 mg / ml. 3 μl / well of labeling buffer (from the HT Protein Charge Variant Labeling Kit), 15 μl / well of test solution and 3 μl / well of Dye Mixture (from the HT Protein Charge Variant Labeling Kit) were introduced into a 96-well plate (Bio-Rad). The plate was placed in the dark for 10 minutes, then 36 μl / well of water was added to each well and mixed by pipetting. The solution was centrifuged at 1000 rpm in an Eppendorf Centrifuge 5417R.

[0295]

[0380] Preparation of working solutions and filling of chips

[0381] The HT Protein Charge Variant Labeling Kit was used to prepare working solutions and chips according to the manufacturer's protocol. The initiation of the assay is a standard procedure. HT Protein Charge Variant 90s was used as the assay method.

[0296]

[0382] Example 9 Determination of sample purity using the Caliper Labchip GXII under reducing and non-reducing conditions

[0383] Preparation of test samples Each denaturing and reducing solution was prepared using 700 μl of HT Protein Express Sample Buffer. For reduced samples, 24.5 μl of 1 M dithiothreitol (DTT) was introduced. For non-reduced samples, 24.5 μl of 1 M iodoacetamide (IAM) was introduced into the sample buffer as an alkylating agent.

[0297]

[0384] For each sample, two microtubes were prepared and 35 μl of denaturation buffer was added to one of the two tubes and 35 μl of reduction buffer was added to the other. Samples were diluted to a concentration of 2 mg / ml. 5 μl of sample was introduced into each pair of tubes. Samples were denatured at 100° C. for 5 minutes. The tubes were vortexed, then 70 μl / tube of water was added and the tubes were vortexed. 44 μl / well of each sample was transferred to wells of a 96-well plate.

[0298]

[0385] Preparation of working solutions and filling of chips The HT Protein Express Reagent Kit was used to prepare working solutions and chips according to the manufacturer's protocol. The initiation of the assay was a standard procedure. HT Protein Express 200 was selected as the assay method.

[0299]

[0386] Example 10 Determination of the charge form profile of samples by ion exchange (IE) high performance liquid chromatography (HPLC) Column: ProPac WCX-10 Analytical, 4 x 250 mm Precolumn: Pro Pac WCX-10G, 4 x 50 mm Column temperature: 30℃ Mobile phase flow rate: 0.7ml / min Injection volume: 50μl Sample concentration: 1mg / ml Detector wavelength: 220nm Elution time: 60 min Mobile phase: Eluent A: 0.03M 2-(N-morpholino)ethanesulfonic acid (MES), pH 6.0 Eluent B: 0.03M MES, 0.5M NaCl, pH6.0 Eluent A gradient: 86%-0%-86%

[0387] Prior to the assay, test samples were treated with carboxypeptidase B for 2 hours at +37°C ± 1°C.

[0300]

[0388] The absolute change in charge form profile after stress was calculated by the following formula: Δ=|acid fraction content before stress−acid fraction content after stress|+|dominant fraction content before stress−dominant fraction content after stress|+|base fraction content before stress−base fraction content after stress.

[0301]

[0389] Example 11 Determination of low molecular weight impurities by reducing and non-reducing vertical gel electrophoresis (VEP) in polyacrylamide gels (PAG).

[0390] Polyacrylamide gels (PAAG) were prepared in glass plates in the presence of sodium dodecyl sulfate, the plates consisting of a concentrating layer of 4% PAAG and a separating layer of 12.5% ​​PAG (under reducing conditions) / 8% PAG (under non-reducing conditions).

[0302]

[0391] The electrophoresis chamber was assembled and set up according to the user manual of the vertical electrophoresis device. The probes were prepared by diluting the samples with purified water to a final concentration of 1 mg / ml. A volume equivalent to 40 μg was taken and the prepared probes of the test samples were mixed with 4x sample buffer solution containing 2-mercaptoethanol (reducing conditions) and 4x sample buffer solution without 2-mercaptoethanol (non-reducing conditions) in a ratio of 3:1 (volume / volume) and stirred. The resulting solutions were incubated at (99±1) °C for 3 min (samples containing 2-mercaptoethanol) and at (99±1) °C for 1 min (samples without 2-mercaptoethanol). The solutions were cooled to room temperature, mixed, and transferred to the PAG wells under the electrode buffer solution layer.

[0303]

[0392] Electrophoresis was performed in constant current mode with a water cooling system. The parameters of the power supply were set: the voltage was 110 V during the passage of the dye front through the stacking gel. After moving the dye front into the low separation gel at a level of 5-7 mm, the voltage was increased to 180 V and the power was turned off when the dye front reached the gel bottom line.

[0304]

[0393] After electrophoresis, the gel was separated from the glass and the proteins were fixed in fixation solution for 16-18 hours at room temperature. The gel was then stained (in Acid Blue 83 solution) and washed to obtain clear visualization of the bands. The gel was scanned. Purity and impurities in the test samples were assessed using GelPro software.

[0305]

[0394] Example 12 Determination of relative specific activity

[0395] The relative specific activity of monoclonal anti-PD-1 antibody samples was assessed by their ability to specifically bind to PD-1 protein on the surface of Jurkat PD-1 NFAT system cell membranes. The day before the analysis, PDL-1 was immobilized on the wells of the culture plate. The next day, the plate was washed and 50 μl / well of phytohemagglutinin P (PanEco, Russia, Cat. No. M021) solution was added. Serial dilutions of standard and test samples were prepared using the Freedom Evo robot and introduced into the culture plate at 10 μl / well. Jurkat PD-1 NFAT cell suspension was introduced into the culture plate at 40 μl / well. The plate was incubated at 37 °C and 5% CO2 for 4-6 h. All the above procedures were performed under sterile conditions.

[0306]

[0396] After incubation, 100 μl / well of BioGlo (Promega, USA, Cat. No. G7941) solution was added and luminescence levels were determined.

[0397] Using Magellan software, we constructed a four-parameter curve for the dependence of the mean luminescence value on the protein concentration of solutions of standards and test samples placed on the same tablet.

[0307]

[0398] The relative specific activity (RP) of the test sample was calculated by the following formula:

[0308]

number

[0309] During the ceremony, ED 50 st is the median effective dose value of the standard sample (ng / ml); ED 50 The test is the median effective amount value (ng / ml) of the test sample.

[0310]

[0399] The mean value of the relative specific activity was taken as the final result and was calculated from three independent measurements (determined from three different culture plates).

[0400] Example 13 Viscosity determination The dynamic viscosity of the test solutions was measured by rotational viscometry using a CAP2000+L Brookfield viscometer.

[0311]

[0401] Example 14 Research into the origins of buffer solutions

[0402] Test preparation Four buffer solutions were selected for this study. The molarity and pH of the buffers were selected taking into account possible subcutaneous administration limitations and the рН of the antibodies (рН of the test solutions was selected at the lowest possible physiological value). Composition (per 1 mL):

[0312] [Table 24]

[0313]

[0403] Determining colloidal stability by PEG aggregation The study was performed in triplicate for each sample, and the results are shown in Table 1 and Figure 1.

[0404]

[0314] [Table 25]

[0315]

[0405] As a result, samples in histidine and acetate buffer solutions showed the highest colloidal stability in the presence of PEG. Phosphate and citrate based compositions were excluded from further studies since aggregation at 6% PEG is an indication of unsatisfactory colloidal stability. Based on the results obtained, acetate and histidine buffer solutions were selected for further pH / molarity selection.

[0316]

[0406] Example 15 Solution pH / Buffer Volume Selection

[0407] Test preparation (per 1 ml)

[0317] [Table 26]

[0318]

[0408] The study was performed by heat stress at 50°C for 72 hours. The results are shown in Table 2. The homogeneity of the samples was determined by SE HPLC and electrophoresis using the Labchip system. The charge form profile was analyzed using the Labchip system.

[0319]

[0409]

[0320] [Table 27-1]

[0321] [Table 27-2]

[0322]

[0410] The results of this study show that the presence of a monoclonal anti-PD-1 antibody (prorugolimab) in aqueous solution leads to an increase in the pH level, which is best stabilized by a 20 mM buffer solution.

[0323]

[0411] All tested samples showed high aggregation stability in SE HPLC, as the increase in impurities after heat stress was less than 0.5% in all samples. The histidine-based samples at pH 5.5-6.0 and the acetic acid-based samples at pH 5.0-5.5 showed the best stability.

[0324]

[0412] All formulations exhibited similar stability of charge form profile, and quantitative differences in fractional content changes between formulations were within the precision limits of the method.

[0413] Reducing gel electrophoresis results showed that all histidine-based formulations showed increased stability, whereas under non-reducing conditions, the acetate-based solution showed the best results.

[0325]

[0414] Based on the obtained data on the stability of pH, purity and charge form profile of monoclonal anti-PD-1 antibodies, the following formulation excipients are recommended for further studies:

[0326] [Table 28]

[0327]

[0415] Example 16 Selection of Histidine-Based Pharmaceutical Compositions

[0416] Test preparation For screening of stable pharmaceutical compositions based on 20 mM histidine buffer solution at pH 5.5, the following excipients were selected: mannitol, trehalose dihydrate, sucrose (osmotic agent). All test solutions were isotonic.

[0328]

[0417] Sample (mg~1ml)

[0329] [Table 29-1]

[0330] [Table 29-2]

[0331]

[0418] Sample stability was evaluated by heat stress at 50°C for 72 hours, shaking test at a speed of 800 rpm for 120 hours, and one-time freezing at -16-20°C and thawing at +25±1°C. The turbidity of the solutions was evaluated by spectrophotometry at 400 nm. The homogeneity of the samples was determined by electrophoresis using SE HPLC and Labchip system. The charge form profile was analyzed using the Labchip system.

[0332]

[0419]

[0333] [Table 30-1]

[0334] [Table 30-2]

[0335] [Table 30-3]

[0336] [Table 30-4]

[0337] [Table 30-5]

[0338]

[0420]

[0339] [Table 31]

[0340]

[0421] Table 3 shows the negative effect of mannitol on the thermal and colloidal stability of monoclonal anti-PD-1 antibody in 20 mM histidine buffer solution at pH 5.5: during 96 hours of heat stress, the increase in impurities by SE HPLC was 1.24%-3.18%, and during 120 hours of shaking tests, the solution showed visible aggregation. The mannitol-containing formulation also showed negative stability results during freeze-concentration, after one cycle of freeze-thaw, the increase in impurities by SE HPLC was 1.05-4.45%, much higher than the other formulations.

[0341]

[0422] The formulations containing L-proline also showed poor colloidal stability as a result of visible aggregation during shaking tests.After freeze-thaw and heat stress tests, the increase in impurities in the formulations containing L-proline averaged more than 1% by SE HPLC.

[0342]

[0423] During the course of the experiments, we detected high thermal and colloidal stability (lack of significant changes in quality indicators against all stresses) of prorugolimab in formulations containing trehalose dihydrate without solubilizers such as polysorbate 80 and poloxamer 188.

[0343]

[0424] The best stabilization effect of monoclonal anti-PD-1 antibody in 20 mM histidine buffer solution was shown by trehalose dihydrate, sucrose, and their combination with glycine. These formulations can be used as lyophilized dosage forms of monoclonal antibodies.

[0344]

[0425] Assessment of the stability of candidate histidine-based formulations after lyophilization

[0426] The samples that showed the best stability in the preliminary screening stage were tested for the possibility of producing a lyophilizate to prepare an infusion solution.

[0345]

[0427] For this purpose, solutions containing 20 mg / ml or 100 mg / ml of the monoclonal anti-PD-1 antibody prorugolimab were placed in hydrolysis class I glass vials and the vials were loosely capped with rubber slotted stoppers. The vials containing the solutions were placed in a freeze-drying chamber. Freeze-drying was performed in automatic mode. The freezing stage was performed at minus 40 °C, primary drying was at (0.10 ± 0.03) mbar, and during secondary drying the temperature was increased and the pressure was at (0.05 ± 0.02) mbar. After the drying step, the pressure was reduced to the required level for vacuum -0.76 ± 0.03 mbar, the vials were capped with rubber stoppers, and then the vacuum was released to atmospheric pressure. The capped vials were transferred for further studies. The freeze-dried samples of the monoclonal anti-PD-1 antibody prorugolimab were stored at 2-8 °C.

[0346]

[0428] To prove the stability of the protein, the samples were reconstituted after lyophilization. The monomer content was evaluated by SE HPLC and the charge form profile was analyzed using the Labchip system. We also measured the pH value before and after lyophilization. The results are shown in Table 5.

[0347]

[0429] A slight shift in pH was observed during lyophilization of selected histidine-based formulations and after lyophilizate reconstitution, which is within the precision limits of the method. All formulations showed high purity stability by SE HPLC and charge form profile after reconstitution. However, gel electrophoresis on the LabChip Caliper system showed significant differences in purity between samples. The most stable samples with regard to this parameter are formulations no. 22, 27, and 36, which contain trehalose dihydrate. Formulations containing L-proline also showed significant changes in the charge form profile. Therefore, they are not recommended for use. The possibility of lyophilization of formulations no. 22 and 27 with a protein concentration of 100 mg / ml was demonstrated.

[0348]

[0430]

[0349] [Table 32-1]

[0350] [Table 32-2]

[0351]

[0431] Preparation of high concentration forms and confirmation of stability under accelerated aging

[0432] Based on the results of screening liquid and lyophilized dosage forms, the following formulations were selected for accelerated stability studies.

[0352] [Table 33]

[0353]

[0433] The trehalose dihydrate content was reduced to equilibrate the monoclonal anti-PD-1 antibody-containing solution with an increased concentration of osmolality to physiological levels (approximately 300 mOsm / kg) and reduce the viscosity of the solution to a level below 100 cP. The stability of the samples was confirmed by accelerated aging at +37°C, and the tests were performed by SE HPLC, IE HPLC and VEP, and the relative specific activity of prorugolimab was also determined. The results are shown in Tables 6 and 7.

[0354]

[0434]

[0355] [Table 34]

[0356]

[0435]

[0357] [Table 35]

[0358]

[0436] Example 17 Selection of Acetate-Based Pharmaceutical Compositions

[0437] Test preparation For screening of stable pharmaceutical compositions based on 20 mM acetate buffer solution at pH 5.0, the following excipients were selected: mannitol, trehalose dihydrate, sucrose (osmotic agent), L-proline (osmotic agent and stabilizer), glycine (osmotic agent and stabilizer), polysorbate 80 and poloxamer P188 (solubilizer). All test solutions were isotonic.

[0359]

[0438] Test preparation (per 1 ml)

[0360] [Table 36]

[0361]

[0439] The stability of the samples was evaluated by heat stress at 50°C for 72 hours, shaking test at 800 rpm for 120 hours, freezing once at -16-20°C and thawing at +25±1°C. The turbidity of the solutions was evaluated by spectrophotometry at 400 nm. The homogeneity of the samples was determined by electrophoresis using SE HPLC and Labchip system. The charge form profile was analyzed using the Labchip system.

[0362]

[0440]

[0363] [Table 37-1]

[0364] [Table 37-2]

[0365]

[0441]

[0366] [Table 38]

[0367]

[0442] This study showed a negative effect of mannitol on the thermal and colloidal stability of a monoclonal anti-PD-1 antibody in 20 mM acetate buffer at pH 5.0: during 96 h of heat stress, the increase in impurities by SE HPLC was 0.48-2.59%; during 120 h of shaking test, all test solutions showed no visible aggregation (see UV spectrophotometric results). Mannitol-containing formulations also showed negative stability during freeze concentration: after one cycle of freeze-thaw, the increase in impurities by SE HPLC was 0.69-9.44%, much higher than other formulations.

[0368]

[0443] Formulations containing L-proline also showed lower thermal stability in terms of purity in SE HPLC analysis: the increase in impurities after 96 hours of stress was 1.64-1.93%. After freeze-thawing, the increase in impurities in formulations containing L-proline was below the average value of the most stable formulation in the group. During the course of the experiment, the solubilizer did not show any effect on the thermal and colloidal stability of the protein. In formulations containing trehalose dihydrate, prorugolimab showed high stability in all stress experiments in the absence of surfactants such as polysorbate 80 or poloxamer 188.

[0369]

[0444] The best stabilization effect of monoclonal anti-PD-1 antibody in 20 mM acetate buffer solution was shown by trehalose dihydrate, sucrose, and their combination with glycine. These formulations can be used to prepare lyophilized dosage forms of monoclonal antibodies.

[0370]

[0445] Determining the stability of candidate acetate-based formulations after lyophilization

[0446] The samples with the best stability in the pre-screening stage were tested for the possibility of producing lyophilizates to prepare infusion solutions.

[0371]

[0447] For this purpose, solutions containing 20 mg / ml or 100 mg / ml of monoclonal anti-PD-1 antibody were placed in hydrolysis class I glass vials and the vials were loosely capped with rubber slotted stoppers. The vials containing the solutions were placed in a freeze-drying chamber. Freeze-drying was performed in automatic mode. The freezing stage was performed at minus 40 °C, primary drying at (0.10 ± 0.03) mbar, and during secondary drying the temperature was increased and the pressure was (0.05 ± 0.02) mbar. After the drying step, the pressure was reduced to the level required for vacuum -0.76 ± 0.03 mbar, the vials were capped with rubber stoppers, and then the vacuum was released to atmospheric pressure. The capped vials were transferred for further studies. The freeze-dried samples of monoclonal anti-PD-1 antibody were stored at 2-8 °C.

[0372]

[0448] To prove the stability of the protein, the samples were reconstituted after lyophilization. The monomer content was evaluated by SE HPLC, and the charge form profile was analyzed using the Labchip system. We also measured the pH value before and after lyophilization. The results are shown in Table 8.

[0373]

[0449] Significant shifts in pH were detected within the range of 0.26-0.45 during lyophilization of selected acetate-based formulations and after lyophilizate reconstitution. Such shifts may be caused by the loss of acetate ions during lyophilization. Therefore, if the above events are observed, acetate-based formulations may be recommended for lyophilization.

[0374]

[0450] All formulations showed high purity stability by SE HPLC and showed charge form profile after reconstitution.Studies showed the feasibility of lyophilization of formulations no. 10 and 12, with protein concentration of 100mg / ml.

[0375]

[0451]

[0376] [Table 39]

[0377]

[0452] Preparation of high concentration forms and confirmation of stability under accelerated aging

[0453] Based on the results of screening liquid and lyophilized dosage forms, the following formulations were selected for accelerated stability studies.

[0378] [Table 40]

[0379]

[0454] The trehalose dihydrate content was reduced, the osmolality level of the solution containing the monoclonal anti-PD-1 antibody was equilibrated with a concentration that increased the osmolality level to physiological levels (approximately 300 mOsm / kg), and the viscosity of the solution was reduced to a level below 100 cP. The samples were aged at +37°C and analyzed by SE HPLC, IE HPLC and VEP; the relative specific activity of the protein was also determined. The results are shown in Tables 9 and 10.

[0380]

[0455]

[0381] [Table 41]

[0382]

[0456]

[0383] [Table 42]

[0384]

[0457] While the invention has been described with reference to preferred embodiments, it should be understood that modifications may be employed, as will be apparent to those skilled in the art. Such modifications and variations are to be considered within the purview and scope of the invention.

[0385]

[0458] Representative non-limiting examples of the present invention have been described in detail above. This detailed description is intended only to teach those skilled in the art further details for implementing preferred aspects of the present teachings, and is not intended to limit the scope of the present invention. Furthermore, each of the additional features and teachings disclosed above and below can be utilized separately or in combination with other features and teachings.

[0386]

[0459] Moreover, the combinations of features and steps disclosed in the above detailed description and experimental examples are not necessary to practice the invention in its broadest sense, but are instead taught to illustrate particular embodiments of the invention. Moreover, the features of the specific embodiments described above, and the corresponding independent and dependent claims below, can be combined in ways not specifically and explicitly recited to provide further useful aspects of the invention.

[0387]

[0460] Further examples of studies using various aqueous pharmaceutical compositions comprising the anti-PD-1 antibody prorugolimab to treat malignant neoplasms such as melanoma, including inoperable or metastatic melanoma, early stage melanoma before and after curative treatment; lung cancer, non-small cell lung cancer, including inoperable or metastatic non-small cell lung cancer (NSCLC), are provided. Exemplary pharmaceutical compositions used in these studies are characterized by the following composition (Table 12.1):

[0388] [Table 43]

[0389]

[0461] Key information regarding clinical development of test products The pharmacokinetics, pharmacodynamics, safety and immunogenicity of BCD-100 (prorugolimab) as monotherapy after intravenous administration were studied in a phase I dose-escalation clinical trial. The study demonstrated the safety and benefit of the test product in patients with common forms of malignant neoplasms (melanoma, NSCLC) of various localizations. Based on the results of this study, two BCD-100 dosing regimens were selected for further clinical development: 3 mg / kg i / v once every 3 weeks and 1 mg / kg i / v once every 2 weeks.

[0390]

[0462] The objective of Study No. BCD-100-2 / MIRACULUM on the Use of BCD-100 in Patients with Unresectable or Metastatic Melanoma was to evaluate the pharmacokinetics, efficacy, safety and immunogenicity of two dosing regimens as monotherapy: 3 mg / kg i / v once every 3 weeks and 1 mg / kg i / v once every 2 weeks. The primary endpoint (ORR) was evaluated based on the results of an interim analysis at 6 months. BCD-100 showed a favorable safety profile in all dosing regimens. Both dosing regimens showed efficacy in patients with unresectable or metastatic melanoma, with a DC of approximately 60% and an ORR of 30%.

[0391]

[0463] Example 18 Use of prorugolimab (BCD-100) to treat patients with common forms of malignant neoplasms of various localizations, Phase I study, BCD-100-1

[0464] Test Design Study No. BCD-100-1 investigated the pharmacokinetics and tolerability of BCD-100 (prorugolimab). Study No. BCD-100-1 (NCT030500047) was a phase I multicenter, open-label study in patients with solid tumors conducted within the territory of the Russian Federation. The main task of the study was to evaluate the pharmacokinetic and clinical pharmacological parameters of BCD-100. The main characteristics of the BCD-100-1 study are presented in Table 13.

[0392]

[0465]

[0393] [Table 44]

[0394]

[0466] The first patient was given a starting dose of the BCD-100 product (0.3 mg / kg) every 2 weeks. In the absence of dose-limiting toxicity for 4 weeks, patients were subjected to dose escalation to 1 mg / kg every 2 weeks. After the first patient, the study continued under a conventional dose escalation 3+3 design; i.e., in the absence of DLTs, cohorts of 3 patients were enrolled sequentially at the next dose level every 4 weeks. At each dose level, patients were assigned to receive BCD-100 for 85 days (approximately 3 months) or until DLT signs / disease progression were observed.

[0395]

[0467] If a DLT event was observed in one patient at a particular dose level, the next cohort of three patients received the same level of product; thus, a total of six patients began receiving this dose of BCD-100. If a DLT event was observed in two or more patients at a particular dose level, dose escalation and allocation of new patients to the product was halted.

[0396]

[0468] The follow-up period for patients after treatment with the BCD-100 product was 126 days. During this period, we monitored adverse events and performed clinical examinations and blood analyses (once every 2 weeks for the first 28 days, then once every 28 days).

[0397]

[0469] Test results Summary of patient population characteristics The study enrolled 15 patients aged 18 years or older with common forms of malignant solid neoplasms of various localizations (melanoma including choroidal melanoma, NSCLC, renal cell carcinoma, mesothelioma) regardless of gender. To be eligible for enrollment, patients had to have an ECOG score of 0-2 according to RECIST 1.1 (excluding bone metastases) and at least one measurable target lesion. Disease characteristics and patient distribution are shown in Tables 14 and 15.

[0398]

[0470]

[0399] [Table 45]

[0400]

[0471]

[0401] [Table 46]

[0402]

[0472] The final analysis includes data on 15 patients (6 patients from the BCD-100 1 mg / kg cohort, including 1 patient after dose escalation; 6 patients from the BCD-100 3 mg / kg cohort, and 3 patients from the BCD-100 mg / kg cohort). All patients were treated within the main phase of the study (85 days).

[0403]

[0473] All patient cohorts were well balanced with regard to key demographic characteristics. The gender distribution of patients was even: 53.33% female (8 patients) and 46.67% male (7 patients). The median age of patients was 56 years (minimum age 35 years, maximum age 77 years).

[0404]

[0474] Regarding main disease characteristics, melanoma patients (9 / 15 patients) predominated in the population. This population also included 4 patients with NSCLC, 1 patient with pleural mesothelioma, and 1 patient with renal cell carcinoma. The median disease duration at screening for the entire population was 17.07 months, with a minimum of 0.6 months and a maximum of 91.2 months.

[0405]

[0475] Summary of clinical safety results from Phase I studies Safety was studied in a population that included all patients (n=15) who received at least one dose of the study product. Each of the 15 patients exhibited AEs and / or SAEs, totaling 247 events. Statistical analysis showed no differences between cohorts with regard to AEs (p=0.567, Kruskal-Wallis test). Table 16 summarises the product safety data and is presented below.

[0406]

[0476] Grade 3 AEs by CTCAE 4.03 were detected in 40.00% (6 / 15) of patients, i.e., 2 patients in each cohort. Investigators determined that grade 3 or higher AEs were treatment-related in 4 of these 6 patients: one patient who received BCD-100 at a dose of 1 mg / kg, one patient who received BCD-100 at a dose of 3 mg / kg, and two patients who received BCD-100 at a dose of 10 mg / kg. The majority of these adverse events were hematologic in nature.

[0407]

[0477] SAEs were observed in three patients receiving BCD-100 at a dose of 1 mg / kg and one patient receiving BCD-100 at a dose of 3 mg / kg.

[0478] One case of dose-limiting toxicity (DLT) was detected during the study. Patient 13-09, who received two doses of BCD-100 at 3 mg / kg, showed an abnormal endocrine profile (reduced TSH) and developed autoimmune thyroid disease (immune-mediated grade 2 adverse event according to CTCAE 4.03 criteria). The Data Monitoring and Safety Committee considered this case a DLT event. Patient 13-09, who continued on study treatment, was classified as having a stable disease course according to RECIST 1.1 and irRC criteria.

[0408]

[0479] Excluding autoimmune thyroid disease, immune-mediated AEs were observed in 33.33% (5 of 15) of patients, 33.33% (2 of 6) of patients receiving BCD-100 1 mg / kg, 50.00% (3 of 6) of patients receiving BCD-100 3 mg / kg, but none of patients receiving BCD-100 10 mg / kg. All of these AEs were grade 1 events according to CTCAE 4.03 and were not considered DLT events.

[0409]

[0480] AEs / SAEs led to temporary discontinuation of study treatment in three patients: 16.67% (1 of 6) in patients receiving 1 mg / kg BCD-100 and 33.33% (2 of 6) in patients receiving 3 mg / kg BCD-100. All AEs that led to temporary treatment discontinuation were immune-related (severity 1 hyperthyroidism in patient 10-03 who received 1 mg / kg BCD-100; severity 1 autoimmune thyroid disease in patient 13-10; and severity 2 autoimmune thyroid disease in patient 13-09 (both patients received 3 mg / kg BCD-100)). Patient 10-03 discontinued study treatment. Patients 13-09 and 13-10 received glucocorticoids for disease and then resumed treatment. There were no treatment interruptions beyond the criteria set by the study protocol (2–4 weeks).

[0410]

[0481] Study product was discontinued in 3 patients due to disease progression (2 after 5 doses at 1 mg / kg and 1 after 5 doses at 10 mg / kg). In all cases, the reason for discontinuation was disease progression, not BCD-100 administration.

[0411]

[0482] SAEs led to treatment discontinuation and subsequent death in one study patient. Patient 13-10, who received three doses of 3 mg / kg, experienced a right hemisphere stroke of grade 5 severity. The investigator determined that this AE was possibly related to study treatment.

[0412]

[0483] No differences were observed in terms of changes in laboratory and physiological parameters over time between patient groups. The analysis of these parameters showed isolated instances of statistically significant differences, but these differences were considered to reflect normal natural fluctuations. Deviations detected after biochemical blood tests and blood coagulation tests (increased levels of transaminases, bilirubin values, deviations in blood coagulation indexes, etc.) were, for the most part, typical for the study population (patients with inoperable neoplasms), expected and transient. No further treatment was required, as the parameters normalized without significant consequences.

[0413]

[0484] The results described showed that the safety profile of the BCD-100 product was favorable at all doses studied after intravenous administration.

[0485]

[0414] [Table 47]

[0415]

[0486] immunogenicity Immunogenicity was assessed at screening, day 28 of study treatment, at the end of the main period (day 85), and in patients who continued treatment with the BCD-100 product, immunogenicity was also assessed every 42 days thereafter. Immunogenicity was assessed in two stages as follows: in step 1, we screened serum samples from patients for the presence of binding antibodies (BAB) against the product, and in step 2, we screened BAB titer positive samples for the presence of neutralizing antibodies (NAB).

[0416]

[0487] Immunogenicity was assessed in all patients with analyzable serum samples at screening and on subsequent analysis days (n=15). No samples were detected containing BAB. As no specimens contained BAB, analysis for the presence of NAT was not performed.

[0417]

[0488] Phase I Pharmacokinetic / Metabolic Overview Pharmacokinetic (PK) assays will include data from omitted / lost / spoiled serum specimens for ≤3 patients to measure BCD-100 concentrations following the first dose of product (n=15).

[0418]

[0489] Pharmacokinetics (after the first dose of the product and throughout the entire treatment period) was studied in the following groups: · 1st patient (BCD-100 dose of 0.3 mg / kg) (n=1); · First cohort of patients (BCD-100 dose of 1 mg / kg) (n=5); · A second cohort of patients (BCD-100 dose of 3 mg / kg) (n=6); · Third cohort of patients (BCD-100 dose of 10 mg / kg) (n=3).

[0419]

[0490] The pharmacokinetic studies included evaluation of the standard pharmacokinetic properties (distribution and excretion) of the investigational product following multiple intravenous doses. The analytical results are shown in Table 17.

[0491] Blood samples for pharmacokinetic assays (BCD-100 content in serum) were taken at each dose level in all enrolled patients. The following time points for sample collection were used: before the first dose of product; 30 min, 2 h (±15 min), 4 h (±15 min), 6 h (±15 min), 24 h (±1 h), 48 h (±2 h), 192 h (±8 h), and 336 h (±8 h) after the first dose of product (before the second dose), and before each subsequent dose of product once every two weeks. Blood samples for determining BCD-100 concentration in serum were collected at the same time as samples for immunogenicity analysis (to evaluate the possible effect of antibodies against the product on the pharmacokinetic properties of BCD-100). BCD-100 concentration in serum was determined using a validated ELISA procedure.

[0420]

[0492]

[0421] [Table 48]

[0422]

[0493] All parameters were measured after a single administration of the product, except for the Cmin parameter, which was calculated after six administrations of the product (12 weeks).

[0494] Statistically significant differences between the three cohorts receiving different doses were detected in the following parameters: AUC(0-336h) (p=0.0088, Kruskal-Wallis test); AUMC(0-336h) (p=0.0088, Kruskal-Wallis test); AUC(0-∞) (p=0.0123, Kruskal-Wallis test); Cmax (p=0.0103, Kruskal-Wallis test). A more favorable pharmacokinetic profile was observed with the 3 mg / kg and 10 mg / kg doses compared to the 1 mg / kg dose, but PK, efficacy and safety parameters did not show any association with dose.

[0423]

[0495] Figures 1 and 2 show the BCD-100 product concentration profiles from first dose to week 10 in all dose cohorts (1 mg / kg cohort [n=5], 3 mg / kg cohort [n=6], and 10 mg / kg cohort [n=3]).

[0424]

[0496] Calculation of PK parameters showed that BCD-100 concentrations were directly proportional to the administered dose, reaching a peak between 30 minutes and 6 hours after administration and then gradually decreasing. We did not detect any dependency between the half-life of the administered product and the dose. The values ​​of the T1 / 2 parameters were typical for monoclonal antibodies (12-18 days).

[0425]

[0497] Summary of pharmacodynamic results from Phase I studies Pharmacodynamics were assessed in all patients who had serum samples available at screening and on subsequent analysis days (n=15).

[0426]

[0498] Pharmacodynamics were studied in the following groups: · 1st patient (BCD-100 dose of 0.3 mg / kg) (n=1); · First cohort of patients (BCD-100 dose of 1 mg / kg) (n=5); · A second cohort of patients (BCD-100 dose of 3 mg / kg) (n=6); · Third cohort of patients (BCD-100 dose of 10 mg / kg) (n=3).

[0427]

[0499] The level of PD-1 receptor saturation by the BCD-100 product helps to evaluate the interaction between the investigational product and its target and is the most important pharmacodynamic marker of activity of monoclonal anti-PD-1 antibodies, since PD-1 receptor blockade activates antitumor immune responses. Blood sampling for pharmacodynamic evaluation (% PD-1 receptor saturation by the BCD-100 product) was performed in all patients. Sample collection for PD evaluation was performed before administration of the product, 4 hours and 336 hours after administration (but before the second dose), and before the sixth dose of the product.

[0428]

[0500] The results show high saturation (95%-100%) of the PD-1 receptor by the BCD-100 product at all dose levels and in all cell populations (Table 18).

[0501]

[0429] [Table 49-1]

[0430] [Table 49-2]

[0431]

[0502] Summary of Phase I clinical efficacy results Fourteen of the 15 patients who received BCD-100 were included in the efficacy population. One patient discontinued treatment due to an SAE (death), but tumor response to treatment was not evaluated by CT scan at the time of the patient's death.

[0432]

[0503] Overall response rate (partial response rate + complete response rate) and disease control (stable disease + partial response + complete response) were determined 85 days after treatment with the BCD-100 product. Efficacy assessments performed 85 days after treatment with the BCD-100 product were based on analysis of CT scan results. Tumor response was evaluated by RECIST 1.1 and Immune-Related RECIST (irRECIST) criteria.

[0433]

[0504] According to the irRECIST criteria developed for immunotherapy, disease control was achieved in 28.57% of patients (4 of 14). The overall response rate was 7.14% (1 of 14). No significant differences in response rates were observed between the different patient cohorts (Table 19).

[0434]

[0505]

[0435] [Table 50]

[0436]

[0506] conclusion The analysis of the obtained data clearly indicates a good safety profile of the BCD-100 product used in a wide range of doses from 0.03 to 10.0 mg / kg. The choice of the following two dosing regimens of the BCD-100 product is considered optimal for further clinical studies: 1 mg / kg once every 2 weeks and 3 mg / kg once every 3 weeks.

[0437]

[0507] Example 19 Use of Prorugolimab, BCD-100, Phase II Study, BCD-100-2 / MIRACULUM, to Treat Patients with Unresectable / Metastatic Melanoma

[0508] Test Design Study number BCD-100-2 / MIRACULUM (MIRACULUM, NCT03269565) is a multicenter, open-label, randomized study investigating the pharmacokinetics, efficacy, safety, and immunogenicity of BCD-100 product (prorugolimab) as monotherapy in previously untreated or treated patients with unresectable / metastatic melanoma. BCD-100 product administered intravenously at a dose of 3 mg / kg once every 3 weeks was compared to BCD-100 product administered intravenously at a dose of 1 mg / kg once every 2 weeks. The study is ongoing in the Russian Federation and Belarus. Data after 1 year of treatment were available and analyzed.

[0438]

[0509] Each efficacy parameter was evaluated separately for each group. The primary efficacy endpoint was the overall response rate (partial response rate + complete response rate) by irRECIST in study participants treated with BCD-100 products. The best response to treatment during the above period was considered for calculation of ORR (overall response rate) and disease control. Secondary efficacy endpoints were progression-free survival and overall survival 12 months after the start of treatment, disease control rate (disease stabilization rate + partial response + complete response), time to treatment response, and duration of treatment response (Figure 3, Table 20).

[0439]

[0510]

[0440] [Table 51]

[0441]

[0511] Test results Summary of patient population characteristics

[0512] We enrolled 131 patients aged 18 years or older. The patient population included both men and women with common melanomas, including choroidal melanoma. This population included patients who discontinued before the first dose of BCD-100 or as a result of serious protocol deviations. The modified ITT population, who received at least one dose of BCD-100, included 126 patients. To be eligible for enrollment, patients must have an ECOG score of 0–1 and at least one measurable target lesion according to RECIST 1.1 (excluding bone metastases) (Figure 4, Table 21).

[0442]

[0513]

[0443] [Table 52-1]

[0444] [Table 52-2]

[0445]

[0514] Safety was analyzed in all patients who received at least one dose of study product (the "modified intention to treat" (mITT) population, n=126).

[0515] Efficacy was analyzed in two patient populations: · All patients who received at least one dose of study product (mITT population, n=126); Patients who received at least one dose of study product and were scheduled for at least one dynamic CT examination to assess response (the "per protocol" (PP) population, n=114).

[0446]

[0516] Pharmacokinetic assays included data on all patients who received at least one dose of BCD-100 product with ≤3 omitted / missing / spoiled samples required according to the protocol version in effect at the time of decision (n=125).

[0447]

[0517] Summary of Clinical Efficacy from Study No. BCD-100-2 / MIRACULUM The results of the evaluation of the primary efficacy endpoint (ORR) show sufficient efficacy in both dose regimens of the BCD-100 product in the entire patient population studied. Thus, the ORR and disease control in the PP population were 40.68% and 67.80% in group 1 and 32.73% and 52.73% in group 2. Similar efficacy results were observed in the mITT population. Thus, the expected target response rate (28%) and critical value r (11 responses) were reached in both treatment groups.

[0448]

[0518] Subgroup analysis of lines of treatment demonstrated high efficacy of the BCD-100 product at the lowest dose of 1 mg / kg in the treatment-naïve patient population (ORR was 50.00%).

[0519] Results from the analysis of secondary efficacy endpoints confirmed conclusions regarding the sufficient efficacy of the BCD-100 product at both doses. PFS, OS, and duration of response rates were comparable to those of best-in-class PD-1 inhibitors.

[0449]

[0520] The 12-month PFS was 41.27% for patients in the BCD-100 1 mg / kg group and 34.92% for patients in the BCD-100 3 mg / kg group. The median PFS was 5.78 months (95%CI 3.52--) for patients in the BCD-100 1 mg / kg group and 2.33 months (95%CI 2.07-10.25) for patients in the BCD-100 3 mg / kg group (p=0.400, log-rank test). No statistically significant differences were observed after detailed evaluation of progression-free survival in each group according to line of treatment (by irRECIST criteria, mITT population). BCD-100 products showed comparable efficacy both when used in previously untreated and previously treated patients.

[0450]

[0521] With a median follow-up of 13.8 months (95% CI 13.2-14.7), the median overall survival rate for patients in the BCD-100 1 mg / kg group had not been reached (95% CI - 1 - -). 1"-" currently means "not reached" in this specification.) The 12-month OS rate was 74.60% in patients in the BCD-100 1 mg / kg group. The median OS in patients in the BCD-100 3 mg / kg group was 15 months (95%CI 9.99--), with a median follow-up period of 14.5 months (95%CI 13.9-15.2). The 12-month OS in patients in the BCD-100 3 mg / kg group was 53.97%. After detailed evaluation of the overall survival rate of each group according to the line of treatment, no statistically significant differences were observed. The BCD-100 product showed comparable efficacy both when used in previously untreated and previously treated patients. The median OS was not reached in any of the subgroups of patients who received BCD-100 at a dose of 1 mg / kg (p=0.800; log-rank test). Median OS was 16.8 months (95%CI 9.33--) in previously untreated patients and 15 months (95%CI 7.46--) in the BCD-100 product 3 mg / kg group (p=0.900; log-rank test) (Table 22, Table 23, Figure 5, Figure 6, Figure 7, Figure 8).

[0451]

[0522]

[0452] [Table 53]

[0453]

[0523] Summary of Clinical Safety Results from Study No. BCD-100-2 / MIRACULUM BCD-100 demonstrated a favorable safety profile at both dose regimens (Table 23).

[0454]

[0524]

[0455] [Table 54]

[0456]

[0525] Based on the above data, it can be concluded that the results obtained in the study are not inconsistent with the known data on the safety profile of monoclonal anti-PD-1 receptor antibody preparations.With the exception of the amount of SAEs leading to death (significantly higher amounts were observed in patients in the BCD-100 3 mg / kg group), no statistically significant differences in the product safety profile were observed between the doses studied.

[0457]

[0526] immunogenicity Immunogenicity was assessed in all patients with serum samples available for analysis at screening and on subsequent analysis days (n=121). BAB against BAD-100 was not detected in any patient.

[0458]

[0527] Summary Pharmacokinetic and Metabolic Results from Study No. BCD-100-2 / MIRACULUM The analysis showed that a single intravenous administration of 1-3 mg / kg doses of the BCD-100 product linearly increased the concentration of the studied substance in plasma, which was characterized by a uniform increase, and the half-life of the product, regardless of the dose administered in vivo, was characterized by a period of 11.5-17 days, which is typical for IgG immunoglobulins (Figures 9, 10).

[0459]

[0528] A trend towards a dose-dependent increase in the concentration of the study product in plasma (C max and AUC 0-t , SS parameter) remained after multiple dosing of the BCD-100 product, which corresponds to literature data on the PK profiles of other anti-PD-1 antibody products.

[0460]

[0529] As shown in clinical trial number BCD-100-2 / MIRACULUM, sustained C at sufficient levels against the background of the treatment course mixindicates that therapeutic concentrations of the product are maintained when the BCD-100 product is administered at a dose of 1 mg / kg once every 2 weeks and when the BCD-100 product is administered at a dose of 3 mg / kg once every 3 weeks. Therefore, the use of BCD-100 in both dosing regimens is justified from the standpoint of pharmacokinetic properties.

[0461]

[0530] Due to the fact that many patients lacked the typical terminal elimination phase after a single dose, the T 1 / 2 The parameters were difficult to assess.

[0531] Overall, the BCD-100 product is characterized by pharmacokinetic properties that result in a long retention time of the product in vivo, both at 1 mg / kg once every 2 weeks and at 3 mg / kg once every 3 weeks, sufficient to maintain stable therapeutic concentrations against the background of multiple doses. No dose-dependent efficacy / safety was observed.

[0462]

[0532] Summary of pharmacodynamic results from Phase I studies

[0533] The percentage of PD-1 receptor saturation with BCD-100 products in all leukocyte subpopulations did not differ between treatment arms. PD-1 saturation in the populations of activated helper T cells and cytotoxic leukocytes >99% was observed in 33.33% (14 of 42) of patients analyzed for this parameter. No statistically significant differences were observed between treatment arms in 8 patients receiving BCD-100 at 1 mg / kg and in 6 patients receiving BCD-100 at 3 mg / kg.

[0463]

[0534] Ki-67 is a universal marker of cell proliferation due to the fact that it is found only in dividing cells and is degraded 1.5-2 hours after mitosis. The Ki-67 marker is detected in telomeres, centromere regions, and within the nucleolus. During treatment, the percentage of Ki-67-positive cytotoxic T cells increased in both groups, but no significant differences were observed. Analysis of the portion of Ki-67-positive cytotoxic T cells also did not reveal any statistically significant differences between the groups.

[0464]

[0535] During treatment, the percentage of Th9 in the total population of helper T cells increased mainly in group 2 (BCD-100, 3 mg / kg Q3W), but no significant difference was observed, and no significant difference was observed after group comparison.

[0465]

[0536] According to available literature data, an early increase in Th9 levels is associated with an improved response to treatment with nivolumab for melanoma. 3 Nonomura Y,Otsuka A, Hakashima C,Seidel JA,Kitoh A,Dainichi T,Hakajima S,Sawada Y,Matsushita S,Aoki M,Takenouchi T,Fujimura T,Hatta N,Koreeda S,Fukushima S,Honda T,Kabashima K,Peripheral blood Th9 cells are a possible pharmacodynamic biomaker of nivolmab treatment efficacy in metastatic melanoma patients.Oncoimmunology_2016 Oct 18;5(12):e1248327.It should be noted that these results were obtained in a limited number of patients (n=42), who were heterogeneous in terms of gender and characteristics of the presenting disease.

[0466]

[0537] After comparative analysis of the Th9 portion of the total population of helper T cells in subgroups showing different types of response to treatment with BCD-100 products, no statistically significant differences were shown in both cohorts. However, the graphs provided show a trend towards improved response to treatment in patients who initially showed increased Th9 levels (Figures 11, 12).

[0467]

[0538] conclusion

[0539] The available efficacy, safety, and pharmacokinetic data for BCD-100 are sufficient to justify its use at both a 1 mg / kg every 2 weeks and a 3 mg / kg every 3 weeks dosing regimen.

[0468]

[0540] An international, multicenter, open-label study in a patient population with unresectable / metastatic melanoma The randomized Phase II study No. BCD-100-2 / MIRACULUM showed a significant therapeutic benefit compared to the known data on the efficacy of chemotherapy, with results comparable to those of the best existing treatments. Given the efficacy data with a favorable safety profile, as obtained in this study, the use of the BCD-100 product in routine clinical practice meets the benefit-risk balance in the specific patient population. Without being limited thereto, the present invention includes the following aspects. [Aspect 1] 1. An aqueous pharmaceutical composition of an anti-PD-1 antibody, comprising: (a) Prorugolimab at a concentration of 15 mg / ml to 40 mg / ml as an antibody; (b) trehalose dihydrate at a concentration of 80 mg / ml to 110 mg / ml; (c) sodium acetate trihydrate in a concentration between 0.2 mg / ml and 2.5 mg / ml; and (d) Acetic acid in an amount that makes the pH 4.5 to 5.5 1. An aqueous pharmaceutical composition comprising: [Aspect 2] 2. The aqueous pharmaceutical composition of embodiment 1, wherein the prorugolimab is present in a concentration of 20 mg / ml. [Aspect 3] 3. The aqueous pharmaceutical composition according to aspect 1 or 2, wherein the trehalose dihydrate is present in a concentration of 95 mg / ml to 105 mg / ml. [Aspect 4] 4. The aqueous pharmaceutical composition of aspect 3, wherein the trehalose dihydrate is present in a concentration of 100 mg / ml. [Aspect 5] 5. The aqueous pharmaceutical composition of aspect 4, wherein the sodium acetate trihydrate is present in a concentration of from 1.6 mg / ml to 1.9 mg / ml. [Aspect 6] 6. The aqueous pharmaceutical composition of aspect 5, wherein the sodium acetate trihydrate is present in a concentration of 1.7 mg / ml to 1.8 mg / ml. [Aspect 7] 7. The aqueous pharmaceutical composition of embodiment 6, wherein the sodium acetate trihydrate is present in a concentration of 1.742 mg / ml. [Aspect 8] 3. The aqueous pharmaceutical composition according to claim 1 or 2, wherein the pH is adjusted to 5.0 by adding acetic acid. [Aspect 9] 1. An aqueous pharmaceutical composition of an anti-PD-1 antibody, comprising: (a) Prorugolimab at a concentration of 90 mg / ml to 150 mg / ml as an antibody; (b) trehalose dihydrate at a concentration of 50 mg / ml to 110 mg / ml; (c) sodium acetate trihydrate in a concentration between 0.2 mg / ml and 2.5 mg / ml; and (d) Acetic acid in an amount that makes the pH 4.5 to 5.5 1. An aqueous pharmaceutical composition comprising: [Aspect 10] 10. The aqueous pharmaceutical composition of aspect 9, wherein the prorugolimab is present in a concentration of 90 mg / ml to 110 mg / ml. [Aspect 11] 11. The aqueous pharmaceutical composition of embodiment 9 or 10, wherein the prorugolimab is present in a concentration of 100 mg / ml. [Aspect 12] 10. The aqueous pharmaceutical composition according to aspect 9, wherein the trehalose dihydrate is present in a concentration of 75 mg / ml to 85 mg / ml. [Aspect 13] 13. The aqueous pharmaceutical composition of embodiment 12, wherein the trehalose dihydrate is present in a concentration of 80 mg / ml. [Aspect 14] The sodium acetate trihydrate is present in a concentration of 1.6 mg / ml to 1.9 mg / ml. 10. The aqueous pharmaceutical composition according to embodiment 9. [Aspect 15] 15. The aqueous pharmaceutical composition of aspect 14, wherein the sodium acetate trihydrate is present in a concentration of 1.7 mg / ml to 1.8 mg / ml. [Aspect 16] 16. The aqueous pharmaceutical composition of embodiment 15, wherein the sodium acetate trihydrate is present in a concentration of 1.742 mg / ml. [Aspect 17] 9. The aqueous pharmaceutical composition according to aspect 8, wherein the pH is adjusted to 5.0 to 5.5 by adding the acetic acid. [Aspect 18] 1. An aqueous pharmaceutical composition of an anti-PD-1 antibody, comprising: (a) Prorugolimab as an antibody at a concentration of 5 mg / ml to 150 mg / ml; (b) trehalose dihydrate at a concentration of 70 mg / ml to 110 mg / ml; (с) L-histidine in a concentration of 0.2 to 2.5 mg / ml; and (d) L-histidine hydrochloride at a concentration of 0.2 to 3.5 mg / ml 1. An aqueous pharmaceutical composition comprising: [Aspect 19] 19. The aqueous pharmaceutical composition of aspect 18, wherein the prorugolimab is present in a concentration of 15 mg / ml to 40 mg / ml. [Aspect 20] 20. The aqueous pharmaceutical composition of aspect 19, wherein the prorugolimab is present in a concentration of 15 mg / ml to 25 mg / ml. [Aspect 21] 21. The aqueous pharmaceutical composition of embodiment 20, wherein said prorugolimab is present in a concentration of 20 mg / ml. [Aspect 22] 22. The aqueous pharmaceutical composition according to any one of aspects 18 to 21, wherein the trehalose dihydrate is present in a concentration of 95 mg / ml to 105 mg / ml. [Aspect 23] 23. The aqueous pharmaceutical composition according to aspect 22, wherein the trehalose dihydrate is present in a concentration of 100 mg / ml. [Aspect 24] 19. The aqueous pharmaceutical composition of aspect 18, wherein the prorugolimab is present in a concentration of 90 mg / ml to 110 mg / ml. [Aspect 25] 25. The aqueous pharmaceutical composition of embodiment 24, wherein said prorugolimab is present in a concentration of 100 mg / ml. [Aspect 26] 26. The aqueous pharmaceutical composition of aspect 18, 24 or 25, wherein the trehalose dihydrate is present in a concentration of 75 mg / ml to 85 mg / ml. [Aspect 27] 27. The aqueous pharmaceutical composition according to embodiment 26, wherein the trehalose dihydrate is present in a concentration of 80 mg / ml. [Aspect 28] 19. The aqueous pharmaceutical composition of embodiment 18, wherein the L-histidine is present in a concentration of 0.7 mg / ml to 1.0 mg / ml. [Aspect 29] 29. The aqueous pharmaceutical composition of embodiment 28, wherein the L-histidine is present in a concentration of 0.92 mg / ml. [Aspect 30] 19. The aqueous pharmaceutical composition according to aspect 18, wherein the L-histidine hydrochloride is present in a concentration of 2.8 mg / ml to 3.3 mg / ml. [Aspect 31] 31. The aqueous pharmaceutical composition of embodiment 30, wherein the L-histidine hydrochloride is present in a concentration of 2.96 mg / ml. [Aspect 32] 19. The aqueous pharmaceutical composition according to aspect 18, wherein the composition has a pH of 5.5 to 6.5. [Aspect 33] 33. The aqueous pharmaceutical composition according to aspect 32, wherein the composition has a pH of 5.5 to 6.0. [Aspect 34] 20. The aqueous pharmaceutical composition according to embodiment 1 or 18, further comprising a suitable solubilizer. [Aspect 35] 35. The aqueous pharmaceutical composition according to embodiment 34, wherein the solubilizing agent is poloxamer 188. [Aspect 36] 36. The aqueous pharmaceutical composition of embodiment 35, wherein the poloxamer 188 is in an amount greater than 0 mg / ml but less than or equal to 1 mg / ml. [Aspect 37] 1. An aqueous pharmaceutical composition of an anti-PD-1 antibody, comprising: (a) Prorugolimab at a concentration of 20 mg / ml as antibody; (b) trehalose dihydrate at a concentration of 100 mg / ml; (c) sodium acetate trihydrate in a concentration of 0.2 to 2.5 mg / ml; and (d) Acetic acid in an amount that makes the pH 4.5 to 5.5 1. An aqueous pharmaceutical composition comprising: [Aspect 38] (a) Prorugolimab at a concentration of 20 mg / ml as antibody; (b) trehalose dihydrate at a concentration of 100 mg / ml; (c) sodium acetate trihydrate in a concentration of 1.7 mg / ml to 1.8 mg / ml; and (d) Acetic acid in an amount that makes the pH 5.0 38. An aqueous pharmaceutical composition of the anti-PD-1 antibody of embodiment 37 comprising: [Aspect 39] (a) Prorugolimab at a concentration of 20 mg / ml as antibody; (b) trehalose dihydrate at a concentration of 100 mg / ml; (с) sodium acetate trihydrate in a concentration of 1.742 mg / ml; and (d) Acetic acid in an amount that makes the pH 5.0 40. An aqueous pharmaceutical composition of the anti-PD-1 antibody of embodiment 38 comprising: [Aspect 40] 1. An aqueous pharmaceutical composition of an anti-PD-1 antibody, comprising: (a) Prorugolimab at a concentration of 100 mg / ml as antibody; (b) trehalose dihydrate at a concentration of 80 mg / ml; (c) sodium acetate trihydrate in a concentration between 0.2 and 2.5 mg / ml; and (d) Acetic acid in an amount that makes the pH 4.5 to 5.5 1. An aqueous pharmaceutical composition comprising: [Aspect 41] (a) Prorugolimab at a concentration of 100 mg / ml as antibody; (b) trehalose dihydrate at a concentration of 80 mg / ml; (c) sodium acetate trihydrate in a concentration of 1.7 mg / ml to 1.8 mg / ml; and (d) Acetic acid in an amount that makes the pH 5.0 to 5.5 41. An aqueous pharmaceutical composition of the anti-PD-1 antibody of embodiment 40 comprising: [Aspect 42] (a) Prorugolimab at a concentration of 100 mg / ml as antibody; (b) trehalose dihydrate at a concentration of 80 mg / ml; (с) sodium acetate trihydrate in a concentration of 1.742 mg / ml; and (d) Acetic acid in an amount that makes the pH 5.0 to 5.5. 42. An aqueous pharmaceutical composition of the anti-PD-1 antibody of embodiment 41 comprising: [Aspect 43] 1. An aqueous pharmaceutical composition of an anti-PD-1 antibody, comprising: (a) Prorugolimab at a concentration of 20 mg / ml as antibody; (b) trehalose dihydrate at a concentration of 100 mg / ml; (с) L-histidine in a concentration of 0.2 to 2.5 mg / ml; and (d) L-histidine hydrochloride at a concentration of 0.2 to 3.5 mg / ml; Including, (e) An aqueous pharmaceutical composition, wherein the composition has a pH of 5.5 to 6.5. [Aspect 44] (a) Prorugolimab at a concentration of 20 mg / ml as antibody; (b) trehalose dihydrate at a concentration of 100 mg / ml; (с) L-histidine at a concentration of 0.92 mg / ml; and (d) L-histidine hydrochloride at a concentration of 2.96 mg / ml Including, (e) the composition has a pH of 5.5; 44. An aqueous pharmaceutical composition of the anti-PD-1 antibody of embodiment 43. [Aspect 45] 1. An aqueous pharmaceutical composition of an anti-PD-1 antibody, comprising: (a) Prorugolimab at a concentration of 100 mg / ml as antibody; (b) trehalose dihydrate at a concentration of 80 mg / ml; (с) L-histidine in a concentration of 0.2 to 2.5 mg / ml; and (d) L-histidine hydrochloride at a concentration of 0.2 to 3.5 mg / ml Including, (e) An aqueous pharmaceutical composition, wherein the composition has a pH of 5.5 to 6.5. [Aspect 46] (a) Prorugolimab at a concentration of 100 mg / ml as antibody; (b) trehalose dihydrate at a concentration of 80 mg / ml; (с) L-histidine at a concentration of 0.92 mg / ml; and (d) L-histidine hydrochloride at a concentration of 2.96 mg / ml Including, (e) The aqueous pharmaceutical composition of the anti-PD-1 antibody of aspect 45, wherein the composition has a pH of 5.5 to 6.0. [Aspect 47] 1. An aqueous pharmaceutical composition of an anti-PD-1 antibody, comprising: (a) Prorugolimab at a concentration of 20 mg / ml as antibody; (b) trehalose dihydrate at a concentration of 100 mg / ml; (c) sodium acetate trihydrate at a concentration of 1.742 mg / ml; (d) Acetic acid in an amount that makes the pH 5.0 1. An aqueous pharmaceutical composition comprising: [Aspect 48] 1. An aqueous pharmaceutical composition of an anti-PD-1 antibody, comprising: (a) Prorugolimab at a concentration of 20 mg as antibody; (b) trehalose dihydrate at a concentration of 100 mg; (с) sodium acetate trihydrate with a concentration of 1.742 mg; (d) acetic acid in an amount to provide a pH of 5.0; and (e) Water for injection up to 1.0 ml 1. An aqueous pharmaceutical composition comprising: [Aspect 49] 49. The aqueous pharmaceutical composition according to any one of aspects 37 to 48, further comprising a suitable solubilizer. [Aspect 50] 50. The aqueous pharmaceutical composition according to embodiment 49, wherein the solubilizing agent is poloxamer 188. [Aspect 51] 51. The aqueous pharmaceutical composition of embodiment 50, wherein said poloxamer 188 is present in an amount of greater than 0 mg / ml but less than or equal to 1 mg / ml. [Aspect 52] 52. The aqueous pharmaceutical composition according to any one of the preceding aspects, wherein the composition is intended for parenteral administration. [Aspect 53] 53. The aqueous pharmaceutical composition according to embodiment 52, wherein the composition is intended for intravenous, subcutaneous or intramuscular administration. [Aspect 54] 52. The aqueous pharmaceutical composition according to any one of the preceding aspects, wherein the composition is present in a vial. [Aspect 55] 55. The aqueous pharmaceutical composition according to embodiment 54, wherein the vial is a glass vial. [Aspect 56] 55. The aqueous pharmaceutical composition according to aspect 54, wherein the vial has a volume of from 1 ml to 50 ml. [Aspect 57] 55. The aqueous pharmaceutical composition according to embodiment 54, wherein the vial has a volume of 5 ml, 10 ml, 15 ml or 20 ml. [Aspect 58] 52. The aqueous pharmaceutical composition of any one of the preceding aspects, wherein the composition is in a syringe. [Aspect 59] 59. The aqueous pharmaceutical composition according to embodiment 58, wherein the syringe has a capacity of 1 ml. [Aspect 60] 59. The aqueous pharmaceutical composition according to embodiment 58, wherein said syringe has a capacity of 2 ml. [Aspect 61] 52. The aqueous pharmaceutical composition of any one of the preceding aspects, wherein the composition is in a pre-filled syringe. [Aspect 62] 62. The aqueous pharmaceutical composition according to embodiment 61, wherein said pre-filled syringe has a capacity of 1 ml. [Aspect 63] 62. The aqueous pharmaceutical composition according to embodiment 61, wherein said pre-filled syringe has a capacity of 2 ml. [Aspect 64] 64. Use of an aqueous pharmaceutical composition of the anti-PD-1 antibody prorugolimab according to any one of Aspects 1 to 63 for treating a malignant neoplasm in a subject in need of such treatment. [Aspect 65] The malignant neoplasm is melanoma, including inoperable or metastatic melanoma, early stage melanoma before or after definitive treatment; lung cancer, non-small cell lung cancer, including inoperable or metastatic non-small cell lung cancer (NSCLC); non-squamous non-small cell lung cancer, squamous cell lung cancer; inoperable or metastatic small cell 65. The use according to embodiment 64, wherein the tumor is selected from the group comprising: small cell lung cancer, including alveolar lung cancer; early stage lung cancer before or after curative treatment; metastatic cervical cancer, including early stage cervical cancer before or after curative treatment; head and neck tumors, including head and neck squamous cell carcinoma; Hodgkin's lymphoma; gastric and intestinal tumors, metastatic squamous cell esophageal cancer; metastatic urothelial carcinoma, bladder cancer, including renal cancer; metastatic endometrial cancer, including early stage endometrial cancer before or after curative treatment; metastatic breast cancer, including early stage endometrial cancer before or after curative treatment; metastatic or inoperable liver cancer, including early stage liver cancer before or after curative treatment; inoperable or metastatic solid tumors, including inoperable or metastatic solid tumors with signs of microsatellite instability. [Aspect 66] Use of an aqueous pharmaceutical composition of the anti-PD-1 antibody prorugolimab according to any one of aspects 1 to 8, 19 to 22, 37 to 39, and 43 to 44 for treating a malignant neoplasm in a subject in need of such treatment. [Aspect 67] 67. The use of aspect 66, wherein the aqueous pharmaceutical composition is administered at a dose of 1 mg / kg body weight of the anti-PD-1 antibody prorugolimab. [Aspect 68] 67. The use of aspect 66, wherein the aqueous pharmaceutical composition is administered at a dose of 3 mg / kg body weight of the anti-PD-1 antibody prorugolimab. [Aspect 69] The use of embodiment 66, wherein the aqueous pharmaceutical composition is administered every two weeks. [Aspect 70] The use of embodiment 66, wherein the aqueous pharmaceutical composition is administered every three weeks. [Aspect 71] 67. The use of aspect 66, wherein the aqueous pharmaceutical composition is administered at a dose of 1 mg / kg body weight of the anti-PD-1 antibody prorugolimab every two weeks. [Aspect 72] 67. The use of aspect 66, wherein the aqueous pharmaceutical composition is administered at a dose of 3 mg / kg body weight of the anti-PD-1 antibody prorugolimab every three weeks. [Aspect 73] The use according to embodiment 66, wherein the aqueous pharmaceutical composition is administered parenterally. [Aspect 74] 74. The use according to embodiment 73, wherein said parenteral administration is intravenous, subcutaneous or intramuscular administration. [Aspect 75] The use according to embodiment 73, wherein the aqueous pharmaceutical composition is administered intravenously as an infusion. [Aspect 76] The use according to embodiment 66, wherein the malignant neoplasm is melanoma, including inoperable or metastatic melanoma, early stage melanoma before or after curative treatment; lung cancer, non-small cell lung cancer, including inoperable or metastatic non-small cell lung cancer (NSCLC). [Aspect 77] A method for treating a malignant neoplasm in a subject in need thereof, comprising the step of administering a therapeutically effective amount of the aqueous pharmaceutical composition according to any one of aspects 1-8, 19-22, 37-39, and 43-44. [Aspect 78] 78. The method of embodiment 77, wherein the aqueous pharmaceutical composition is administered at a dose of 1 mg / kg body weight of the anti-PD-1 antibody prorugolimab. [Aspect 79] The method of embodiment 77, wherein the aqueous pharmaceutical composition is administered at a dose of 3 mg / kg body weight of the anti-PD-1 antibody prorugolimab. [Aspect 80] The method of embodiment 77, wherein the aqueous pharmaceutical composition is administered every two weeks. [Aspect 81] The method of embodiment 77, wherein the aqueous pharmaceutical composition is administered every three weeks. [Aspect 82] 78. The method of embodiment 77, wherein the aqueous pharmaceutical composition is administered at a dose of 1 mg / kg body weight of the anti-PD-1 antibody prorugolimab every two weeks. [Aspect 83] The method of embodiment 77, wherein the aqueous pharmaceutical composition is administered at a dose of 3 mg / kg body weight of the anti-PD-1 antibody prorugolimab every three weeks. [Aspect 84] The method of embodiment 77, wherein the aqueous pharmaceutical composition is administered parenterally. [Aspect 85] The method of embodiment 84, wherein said parenteral administration is intravenous, subcutaneous, or intramuscular administration. [Aspect 86] The method of embodiment 84, wherein the aqueous pharmaceutical composition is administered intravenously as an infusion. [Aspect 87] The method of embodiment 77, wherein the malignant neoplasm is melanoma, including inoperable or metastatic melanoma, early stage melanoma before or after curative treatment; lung cancer, non-small cell lung cancer, including inoperable or metastatic non-small cell lung cancer (NSCLC). [Aspect 88] 1. A method for treating a malignant neoplasm in a subject in need thereof, comprising administering a therapeutically effective amount of prorugolimab. [Aspect 89] The method of embodiment 88, wherein said prorugolimab is administered at a dose of 1 mg / kg. [Aspect 90] The method of embodiment 88, wherein said prorugolimab is administered at a dose of 3 mg / kg. [Aspect 91] The method of embodiment 88, wherein said prorugolimab is administered every two weeks. [Aspect 92] The method of embodiment 88, wherein said prorugolimab is administered every three weeks. [Aspect 93] The method of embodiment 88, wherein said prorugolimab is administered at a dose of 1 mg / kg every two weeks. [Aspect 94] The method of embodiment 88, wherein said prorugolimab is administered at a dose of 3 mg / kg every three weeks. [Aspect 95] The method of embodiment 88, wherein the prorugolimab is administered parenterally. [Aspect 96] The method of embodiment 95, wherein said parenteral administration is intravenous, subcutaneous, or intramuscular administration. [Aspect 97] The method of embodiment 95, wherein said prorugolimab is administered intravenously as an infusion. [Aspect 98] The method of embodiment 88, wherein the malignant neoplasm is melanoma, including inoperable or metastatic melanoma, early stage melanoma before or after curative treatment; lung cancer, non-small cell lung cancer, including inoperable or metastatic non-small cell lung cancer (NSCLC).

Claims

1. An aqueous pharmaceutical composition of an anti-PD-1 antibody, comprising: (a) prorugolimab at a concentration of 15 mg / ml to 40 mg / ml as the antibody; (b) trehalose dihydrate at a concentration of 80 mg / ml to 110 mg / ml; (c) sodium acetate trihydrate at a concentration of 0.2 mg / ml to 2.5 mg / ml; and (d) acetic acid in an amount sufficient to bring the pH to 4.5 to 5.5 1. An aqueous pharmaceutical composition comprising:

2. 2. The aqueous pharmaceutical composition of claim 1, wherein the prorugolimab is present in a concentration of 20 mg / ml.

3. 3. The aqueous pharmaceutical composition according to claim 1 or 2, wherein the trehalose dihydrate is present in a concentration of from 95 mg / ml to 105 mg / ml.

4. 4. The aqueous pharmaceutical composition of claim 3, wherein the trehalose dihydrate is present in a concentration of 100 mg / ml.

5. 5. The aqueous pharmaceutical composition of claim 4, wherein the sodium acetate trihydrate is present in a concentration of from 1.6 mg / ml to 1.9 mg / ml.

6. 6. The aqueous pharmaceutical composition of claim 5, wherein the sodium acetate trihydrate is present in a concentration of 1.7 mg / ml to 1.8 mg / ml.

7. 7. The aqueous pharmaceutical composition of claim 6, wherein the sodium acetate trihydrate is present in a concentration of 1.742 mg / ml.

8. 3. The aqueous pharmaceutical composition according to claim 1, wherein the pH is adjusted to 5.0 by adding acetic acid.

9. An aqueous pharmaceutical composition of an anti-PD-1 antibody, comprising: (a) prorugolimab at a concentration of 90 mg / ml to 150 mg / ml as the antibody; (b) trehalose dihydrate at a concentration of 50 mg / ml to 110 mg / ml; (c) sodium acetate trihydrate at a concentration of 0.2 mg / ml to 2.5 mg / ml; and (d) acetic acid in an amount sufficient to bring the pH to 4.5 to 5.5 1. An aqueous pharmaceutical composition comprising:

10. 10. The aqueous pharmaceutical composition of claim 9, wherein the prorugolimab is present in a concentration of 90 mg / ml to 110 mg / ml.

11. 11. The aqueous pharmaceutical composition of claim 9 or 10, wherein the prorugolimab is present in a concentration of 100 mg / ml.

12. 10. The aqueous pharmaceutical composition of claim 9, wherein the trehalose dihydrate is present in a concentration of 75 mg / ml to 85 mg / ml.

13. 13. The aqueous pharmaceutical composition of claim 12, wherein the trehalose dihydrate is present in a concentration of 80 mg / ml.

14. The sodium acetate trihydrate is present in a concentration of 1.6 mg / ml to 1.9 mg / ml. The aqueous pharmaceutical composition according to claim 9.

15. 15. The aqueous pharmaceutical composition of claim 14, wherein the sodium acetate trihydrate is present in a concentration of 1.7 mg / ml to 1.8 mg / ml.

16. 16. The aqueous pharmaceutical composition of claim 15, wherein the sodium acetate trihydrate is present in a concentration of 1.742 mg / ml.

17. 10. The aqueous pharmaceutical composition according to claim 9, wherein the pH is adjusted to 5.0 to 5.5 by adding acetic acid.

18. An aqueous pharmaceutical composition of an anti-PD-1 antibody, comprising: (a) prorugolimab at a concentration of 5 mg / ml to 150 mg / ml as the antibody; (b) trehalose dihydrate at a concentration of 70 mg / ml to 110 mg / ml; (c) L-histidine at a concentration of 0.2 to 2.5 mg / ml; and (d) L-histidine hydrochloride at a concentration of 0.2 to 3.5 mg / ml 1. An aqueous pharmaceutical composition comprising:

19. 19. The aqueous pharmaceutical composition of claim 18, wherein the prorugolimab is present in a concentration of 15 mg / ml to 40 mg / ml.

20. 20. The aqueous pharmaceutical composition of claim 19, wherein the prorugolimab is present in a concentration of 15 mg / ml to 25 mg / ml.

21. 21. The aqueous pharmaceutical composition of claim 20, wherein the prorugolimab is present in a concentration of 20 mg / ml.

22. 22. The aqueous pharmaceutical composition according to any one of claims 18 to 21, wherein the trehalose dihydrate is present in a concentration of from 95 mg / ml to 105 mg / ml.

23. 23. The aqueous pharmaceutical composition of claim 22, wherein the trehalose dihydrate is present in a concentration of 100 mg / ml.

24. 19. The aqueous pharmaceutical composition of claim 18, wherein the prorugolimab is present in a concentration of 90 mg / ml to 110 mg / ml.

25. 25. The aqueous pharmaceutical composition of claim 24, wherein the prorugolimab is present in a concentration of 100 mg / ml.

26. 26. The aqueous pharmaceutical composition of claim 18, 24 or 25, wherein the trehalose dihydrate is present in a concentration of from 75 mg / ml to 85 mg / ml.

27. 27. The aqueous pharmaceutical composition of claim 26, wherein the trehalose dihydrate is present in a concentration of 80 mg / ml.

28. 19. The aqueous pharmaceutical composition of claim 18, wherein the L-histidine is present in a concentration of 0.7 mg / ml to 1.0 mg / ml.

29. 29. The aqueous pharmaceutical composition of claim 28, wherein the L-histidine is present in a concentration of 0.92 mg / ml.

30. 19. The aqueous pharmaceutical composition of claim 18, wherein the L-histidine hydrochloride is present in a concentration of from 2.8 mg / ml to 3.3 mg / ml.

31. 31. The aqueous pharmaceutical composition of claim 30, wherein the L-histidine hydrochloride is present in a concentration of 2.96 mg / ml.

32. 19. The aqueous pharmaceutical composition of claim 18, wherein the composition has a pH of 5.5 to 6.

5.

33. 33. The aqueous pharmaceutical composition of claim 32, wherein the composition has a pH of 5.5 to 6.

0.

34. 20. The aqueous pharmaceutical composition of claim 1 or 18, further comprising a suitable solubilizing agent.

35. 35. The aqueous pharmaceutical composition of claim 34, wherein the solubilizing agent is poloxamer 188.

36. 36. The aqueous pharmaceutical composition of claim 35, wherein the poloxamer 188 is in an amount greater than 0 mg / ml but less than or equal to 1 mg / ml.

37. An aqueous pharmaceutical composition of an anti-PD-1 antibody, comprising: (a) prorugolimab at a concentration of 20 mg / ml as the antibody; (b) trehalose dihydrate at a concentration of 100 mg / ml; (c) sodium acetate trihydrate at a concentration of 0.2 to 2.5 mg / ml; and (d) acetic acid in an amount sufficient to bring the pH to 4.5 to 5.5 1. An aqueous pharmaceutical composition comprising:

38. (a) prorugolimab at a concentration of 20 mg / ml as the antibody; (b) trehalose dihydrate at a concentration of 100 mg / ml; (c) sodium acetate trihydrate at a concentration of 1.7 mg / ml to 1.8 mg / ml; and (d) acetic acid in an amount sufficient to bring the pH to 5.0 38. The aqueous pharmaceutical composition of claim 37, comprising the anti-PD-1 antibody.

39. (a) prorugolimab at a concentration of 20 mg / ml as the antibody; (b) trehalose dihydrate at a concentration of 100 mg / ml; (c) sodium acetate trihydrate at a concentration of 1.742 mg / ml; and (d) acetic acid in an amount sufficient to bring the pH to 5.0 The aqueous pharmaceutical composition of claim 38, comprising the anti-PD-1 antibody.

40. An aqueous pharmaceutical composition of an anti-PD-1 antibody, comprising: (a) prorugolimab at a concentration of 100 mg / ml as the antibody; (b) trehalose dihydrate at a concentration of 80 mg / ml; (c) sodium acetate trihydrate at a concentration of 0.2 to 2.5 mg / ml; and (d) acetic acid in an amount sufficient to bring the pH to 4.5 to 5.5 1. An aqueous pharmaceutical composition comprising:

41. (a) prorugolimab at a concentration of 100 mg / ml as the antibody; (b) trehalose dihydrate at a concentration of 80 mg / ml; (c) sodium acetate trihydrate at a concentration of 1.7 mg / ml to 1.8 mg / ml; and (d) acetic acid in an amount sufficient to bring the pH to 5.0 to 5.5 The aqueous pharmaceutical composition of claim 40, comprising:

42. (a) prorugolimab at a concentration of 100 mg / ml as the antibody; (b) trehalose dihydrate at a concentration of 80 mg / ml; (c) sodium acetate trihydrate at a concentration of 1.742 mg / ml; and (d) acetic acid in an amount sufficient to bring the pH to 5.0 to 5.5 42. The aqueous pharmaceutical composition of claim 41, comprising:

43. An aqueous pharmaceutical composition of an anti-PD-1 antibody, comprising: (a) prorugolimab at a concentration of 20 mg / ml as the antibody; (b) trehalose dihydrate at a concentration of 100 mg / ml; (c) L-histidine at a concentration of 0.2 to 2.5 mg / ml; and (d) L-histidine hydrochloride at a concentration of 0.2 to 3.5 mg / ml; Including, (e) An aqueous pharmaceutical composition, wherein the composition has a pH of 5.5 to 6.

5.

44. (a) prorugolimab at a concentration of 20 mg / ml as the antibody; (b) trehalose dihydrate at a concentration of 100 mg / ml; (C) L-histidine at a concentration of 0.92 mg / ml; and (d) L-histidine hydrochloride at a concentration of 2.96 mg / ml Including, (e) the composition has a pH of 5.5; The aqueous pharmaceutical composition of the anti-PD-1 antibody of claim 43.

45. An aqueous pharmaceutical composition of an anti-PD-1 antibody, comprising: (a) prorugolimab at a concentration of 100 mg / ml as the antibody; (b) trehalose dihydrate at a concentration of 80 mg / ml; (c) L-histidine at a concentration of 0.2 to 2.5 mg / ml; and (d) L-histidine hydrochloride at a concentration of 0.2 to 3.5 mg / ml Including, (e) An aqueous pharmaceutical composition, wherein the composition has a pH of 5.5 to 6.

5.

46. (a) prorugolimab at a concentration of 100 mg / ml as the antibody; (b) trehalose dihydrate at a concentration of 80 mg / ml; (C) L-histidine at a concentration of 0.92 mg / ml; and (d) L-histidine hydrochloride at a concentration of 2.96 mg / ml Including, (e) The anti-PD-1 antibody aqueous pharmaceutical composition of claim 45, wherein the composition has a pH of 5.5 to 6.

0.

47. An aqueous pharmaceutical composition of an anti-PD-1 antibody, comprising: (a) prorugolimab at a concentration of 20 mg / ml as the antibody; (b) trehalose dihydrate at a concentration of 100 mg / ml; (c) sodium acetate trihydrate at a concentration of 1.742 mg / ml; (d) acetic acid in an amount sufficient to bring the pH to 5.0 1. An aqueous pharmaceutical composition comprising:

48. An aqueous pharmaceutical composition of an anti-PD-1 antibody, comprising: (a) prorugolimab at a concentration of 20 mg as antibody; (b) trehalose dihydrate at a concentration of 100 mg; (c) sodium acetate trihydrate in a concentration of 1.742 mg; (d) acetic acid in an amount to bring the pH to 5.0; and (e) up to 1.0 ml of water for injection 1. An aqueous pharmaceutical composition comprising:

49. 49. The aqueous pharmaceutical composition according to any one of claims 37 to 48, further comprising a suitable solubilizer.

50. 50. The aqueous pharmaceutical composition of claim 49, wherein the solubilizing agent is poloxamer 188.

51. 51. The aqueous pharmaceutical composition of claim 50, wherein the poloxamer 188 is present in an amount greater than 0 mg / ml but less than or equal to 1 mg / ml.

52. 52. The aqueous pharmaceutical composition of any one of claims 1 to 51, wherein the composition is intended for parenteral administration.

53. 53. The aqueous pharmaceutical composition of claim 52, wherein the composition is intended for intravenous, subcutaneous or intramuscular administration.

54. 52. The aqueous pharmaceutical composition of any one of claims 1 to 51, wherein the composition is present in a vial.

55. 55. The aqueous pharmaceutical composition of claim 54, wherein the vial is a glass vial.

56. 55. The aqueous pharmaceutical composition of claim 54, wherein the vial has a volume of 1 ml to 50 ml.

57. 55. The aqueous pharmaceutical composition of claim 54, wherein the vial has a volume of 5 ml, 10 ml, 15 ml or 20 ml.

58. 52. The aqueous pharmaceutical composition of any one of claims 1 to 51, wherein the composition is in a syringe.

59. 59. The aqueous pharmaceutical composition of claim 58, wherein the syringe has a capacity of 1 ml.

60. 59. The aqueous pharmaceutical composition of claim 58, wherein the syringe has a capacity of 2 ml.

61. 52. The aqueous pharmaceutical composition of any one of claims 1 to 51, wherein the composition is in a pre-filled syringe.

62. 62. The aqueous pharmaceutical composition of claim 61 , wherein the pre-filled syringe has a capacity of 1 ml.

63. 62. The aqueous pharmaceutical composition of claim 61 , wherein the pre-filled syringe has a capacity of 2 ml.

64. An aqueous pharmaceutical composition according to any one of claims 1 to 63 for treating a malignant neoplasm in a subject in need of such treatment.

65. The malignant neoplasm is melanoma, including inoperable or metastatic melanoma, early stage melanoma before or after definitive treatment; lung cancer, non-small cell lung cancer, including inoperable or metastatic non-small cell lung cancer (NSCLC); non-squamous non-small cell lung cancer, squamous cell lung cancer; inoperable or metastatic small cell 65. The aqueous pharmaceutical composition of claim 64, wherein the tumor is selected from the group comprising: small cell lung cancer, including alveolar lung cancer; early stage lung cancer before or after curative treatment; metastatic cervical cancer, including early stage cervical cancer before or after curative treatment; head and neck tumors, including head and neck squamous cell carcinoma; Hodgkin's lymphoma; stomach and intestinal tumors, metastatic squamous cell esophageal cancer; bladder cancer, including metastatic urothelial carcinoma, renal cancer; metastatic endometrial cancer, including early stage endometrial cancer before or after curative treatment; metastatic breast cancer, including early stage endometrial cancer before or after curative treatment; metastatic or inoperable liver cancer, including early stage liver cancer before or after curative treatment; inoperable or metastatic solid tumors, including inoperable or metastatic solid tumors with signs of microsatellite instability.

66. The aqueous pharmaceutical composition of any one of claims 1 to 8, 19 to 22, 37 to 39, and 43 to 44, for treating a malignant neoplasm in a subject in need of such treatment.

67. 67. The aqueous pharmaceutical composition of claim 66, administered at a dose of 1 mg / kg body weight of the anti-PD-1 antibody prorugolimab.

68. 67. The aqueous pharmaceutical composition of claim 66, administered at a dose of 3 mg / kg body weight of the anti-PD-1 antibody prorugolimab.

69. 67. The aqueous pharmaceutical composition of claim 66, which is administered every two weeks.

70. 67. The aqueous pharmaceutical composition of claim 66, which is administered every three weeks.

71. 67. The aqueous pharmaceutical composition of claim 66, wherein the anti-PD-1 antibody prorugolimab is administered at a dose of 1 mg / kg body weight every two weeks.

72. 67. The aqueous pharmaceutical composition of claim 66, wherein the anti-PD-1 antibody prorugolimab is administered at a dose of 3 mg / kg body weight every three weeks.

73. 67. The aqueous pharmaceutical composition of claim 66, which is administered parenterally.

74. 74. The aqueous pharmaceutical composition of claim 73, wherein the parenteral administration is intravenous, subcutaneous or intramuscular administration.

75. 74. The aqueous pharmaceutical composition of claim 73, which is administered intravenously as an infusion.

76. 67. The aqueous pharmaceutical composition of claim 66, wherein the malignant neoplasm is melanoma, including inoperable or metastatic melanoma, early stage melanoma before or after curative treatment; lung cancer, non-small cell lung cancer, including inoperable or metastatic non-small cell lung cancer (NSCLC).

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