Immunomodulatory treatment of body cavities

Abstract

Compositions and methods for treating body cavity cancers, including urinary tract cancers, by a combination of at least one immunomodulatory agent, such as a CTLA-4 antibody or a TLR agonist, and optionally one chemotherapeutic agent, wherein one or more of the therapeutic agents are embedded in a biocompatible hydrogel composition and slowly released from the composition.

Description

Detailed description of the invention 【0001】 [Cross-reference of related applications]

[0001] A benefit is claimed based on U.S. Provisional Patent Application No. 63 / 502,418, filed on 16 May 2023, and the contents of said Provisional Patent Application are incorporated herein by reference in their entirety. 【0002】 [Field]

[0002] Compositions and methods for treating body cavity cancers, including urinary tract cancers, by a combination of at least one immunomodulator and optionally one chemotherapeutic agent, wherein one or more of the therapeutic agents are embedded in a biocompatible hydrogel composition, either in combination or individually, and are slowly released from the composition. 【0003】 [background]

[0003] Cancer is a disease in which abnormal cells proliferate uncontrollably. Urothelial carcinoma of the bladder accounts for more than 90% of all bladder cancers, and of these, about 80% is nonmuscle-invasive bladder cancer (NMIBC), a superficial form of cancer. Transurethral resection and radical cystectomy continue to be the main treatments for NMIBC and MIBC (muscle-invasive bladder cancer), respectively. 【0004】

[0004] The standard treatment for high-grade (HG) NMIBC is transurethral resection of bladder tumor (TURBT) followed by intravesical adjuvant immunotherapy with Bacillus Calmette-Guerin (BCG). Approximately one-third of NMIBC patients do not respond to BCG, and more than half of those who show an initial response experience recurrence or progression during long-term follow-up. For NMIBC patients who do not respond to BCG (i.e., patients with BCG-resistant tumors and BCG-recurrent tumors for which further BCG therapy is not recommended), radical cystectomy is strongly recommended due to the risk of invasive and metastatic disease, and the need for non-surgical treatment options when BCG is unsuccessful is being highlighted. 【0005】 【0005】To activate innate and / or acquired immunity against a target tumor, certain combinations of immunomodulatory compositions and chemotherapeutic compositions have been proposed. For example, anti-CTLA-4 antibodies have been investigated for use in the treatment of various cancers, including bladder cancer. Evaluation of blood samples from patients treated systemically with two doses of ipilimumab (an anti-CTLA-4 antibody) showed an increase in the expression of CD4 + ICOS hi T cells and an approximately five-fold increase in the level of IFN-γ compared to untreated patients, suggesting that ipilimumab increases tumor-responsive cells. However, the toxicity resulting from systemic administration of certain treatments, such as anti-CTLA-4 antibodies, has limited the usefulness of such treatments. 【0006】 【0006】Furthermore, some immune response regulators have a relatively short half-life in terms of their residence time in a body cavity. Such regulators are known to be excreted or metabolized, or simply diffuse out of the body cavity. This short residence duration may reduce the ability of immune checkpoint regulators to activate some immune system cells at the desired site. 【0007】 【0007】Therefore, there remains a great continuing need for new means to control the delivery and activity of immune checkpoint regulators for effective and safe treatment of bladder cancer. 【0008】 [Summary] 【0008】(i) A thermoreversible hydrogel having a viscosity of 5000 cP or less at 5°C and a viscosity of 1×10 6 cP to 9×10 7 cP at 17°C, and (ii) a pharmaceutical composition comprising a mixture with a zarefrelimab liquid solution are described herein. 【0009】

[0009] Further described herein are pharmaceutical compositions for use in the treatment of bladder cancer, wherein a thermoreversible hydrogel is administered intravesically into the bladder of a subject, thereby resulting in a urinary concentration of at least 20-70 μg / mL of zarifremab in the urine of the subject at least 2 hours after administration. 【0010】

[0010] The Specified Specified Kit for Preparing a Pharmaceutical Composition also includes (i) a thermoreversible hydrogel containing 20% ​​to 32% poloxamer 407 and (ii) a solution containing 100 mg to 750 mg of zarifremab, wherein mixing (i) and (ii) forms about 20 to 100 mL of the pharmaceutical composition. 【0011】

[0011] An additional kit for preparing a pharmaceutical composition is described herein, comprising (i) 60 to 73 mL of a thermoreversible hydrogel containing 20% ​​to 32% poloxamer 407 and (ii) 2 to 15 mL of a solution containing 100 mg to 750 mg of zarifremab, wherein mixing (i) and (ii) forms approximately 75 mL of the pharmaceutical composition. 【0012】

[0012] The Specified Specified Kit for Preparing a Pharmaceutical Composition also comprises (i) about 40 to 80 mL of a thermoreversible hydrogel, (ii) about 1 to 10 mL of a mixture of a stabilizer, an active oxygen scavenger, a surfactant, a buffer, and a gelling temperature regulator, and (iii) about 10 to 20 mL of a solution containing 50 mg to 750 mg of zarifremab, wherein when (i), (ii), and (iii) are mixed, about 50 to 100 mL of the pharmaceutical composition is formed. 【0013】

[0013] A ready-to-use pharmaceutical composition for administration to a patient, comprising zarifremab, poloxamer, stabilizers, buffers, reactive oxygen species scavengers, and gelling temperature regulators, is further described herein. 【0014】

[0014] The above and other purposes, features and advantages will become more apparent from the following detailed description, which will proceed with reference to the attached drawings. [Brief explanation of the drawing] 【0015】 [Figure 1] The mean (with 95% confidence interval) urinary concentration of UGN-301 as a percentage of nominal time, for treatment groups and nominal time at concentrations of 100 mg, 300 mg, and 500 mg. Detailed explanation 【0016】

[0016] Terminology 【0017】

[0017] Unless otherwise specified, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art of the field to which this disclosure belongs. The singular terms “a,” “an,” and “the” include plural references unless the context clearly indicates otherwise. Similarly, the term “or” includes “and” unless the context clearly indicates otherwise. Naturally, all base sizes or amino acid sizes, and all molecular weight or molecular mass values ​​given to nucleic acids, polypeptides, and small molecules are approximations and are given for illustrative purposes. Methods and materials similar to or equivalent to those described herein may be used in the practice or testing of this disclosure, but suitable methods and materials are described below. The term “comprise” means “include.” “Substantially consisting of” means a composition, method, or process that includes only the listed features as active or essential elements, but may further include inactive elements. The abbreviation "eg" originates from the Latin phrase "exempli gratia" and is used herein to indicate non-restrictive examples. Therefore, the abbreviation "eg" is synonymous with the term "for example." 【0018】

[0018] In the event of any inconsistency, this specification shall prevail, including any definitions of terms. In addition, all materials, methods, and examples are for illustrative purposes only and shall not be limiting. 【0019】

[0019] Administration: Introduction of the composition to the target via a selected route. The administration of the active compound or composition can be carried out by any route known to those skilled in the art and appropriate to the compound and delivery system. For example, the composition for use in the method described is typically administered locally to the inner surface of a body cavity, such as by intravesical instillation (an exemplary form of local administration). Additional local administration routes may be retrograde administration, such as toward the upper urinary tract. Further examples of local administration include, but are not limited to, local administration, subcutaneous administration, intramuscular administration, subarachnoid administration, intrapericardial administration, intraocular administration, local ophthalmic administration, or administration to the nasal mucosa or lungs by inhalation. In addition, local administration includes administration routes typically used for systemic administration, such as intravascular administration directed toward arterial supply to a specific organ. Thus, in certain embodiments, local administration includes intra-arterial and intravenous administration, where such intra-arterial and intravenous administrations target the vascular system that supplies a specific organ. Local administration also includes incorporating the active compound and agonist into an implantable device or structure, such as a vascular stent or other reservoir, that releases the active agonist and compound over a long period of time for sustained therapeutic effect. 【0020】

[0020] Agonist: A molecule or compound that binds to a target and stimulates a biological response similar to the biological response of a natural substance that normally binds to that target. The target can be a receptor. Agonists are not limited to a specific type of compound and may include peptides and their fragments, as well as other organic or inorganic compounds (e.g., peptide mimes and small molecules) in various embodiments. Agonists may be endogenous, exogenous, complete, partial, reverse, irreversible, or selective agonists. 【0021】

[0021] Analogues, derivatives, or mimetic compounds: Analogues are molecules that differ from the parent compound in chemical structure, such as homologues (differentiated by increments in chemical structure, such as differences in alkyl chain length), molecular fragments, structures that differ by one or more functional groups, or changes in ionization. Structural analogues are often found using quantitative structure-activity relationships (QSAR) with techniques such as those disclosed in Remington (The Science and Practice of Pharmacology, 19th edition (1995), Chapter 28). Derivatives are biologically active molecules derived from a basic structure and include “functional derivatives” as described herein. Mimetic compounds are molecules that mimic the activity of another molecule, such as a biologically active molecule. A biologically active molecule may include a chemical structure that mimics the biological activity of a compound. It is recognized that these terms may overlap under certain circumstances. In certain embodiments of the claimed method, analogues, derivatives, or mimetic compounds having equivalent activity to the explicitly listed compounds may be used in place of the listed compounds. 【0022】

[0022] Animals: Living multicellular vertebrate organisms, a category that includes, for example, mammals and birds. The term mammal includes both humans and non-human mammals. Similarly, the term subject includes both human subjects and veterinary subjects, for example, humans, non-human primates, dogs, cats, horses, and cattle. The term "primate" includes both humans and non-human primates. "Non-human primates" are ape primates such as monkeys, chimpanzees, orangutans, baboons, and macaques. Similarly, the term "subject" includes both human subjects and veterinary subjects such as non-human primates that have internal cavities in which the described method may be effective. 【0023】

[0023] Antagonist: A molecule or compound that tends to neutralize the action of another molecule or compound, or, in some examples, a molecule or compound that blocks a biological response by blocking the ability of a certain chemical substance to bind to its own receptor or other interacting molecule. Antagonists are not limited to a specific type of compound and may include peptides, antibodies and their fragments, and other organic or inorganic compounds (e.g., peptide mimes and small molecules) in various embodiments. In certain embodiments, the antagonist compound is a type of regulatory compound. 【0024】

[0024] Antibody: A protein (or protein complex) comprising one or more polypeptides substantially encoded by an immunoglobulin gene or a fragment of an immunoglobulin gene. Recognized immunoglobulin genes include the constant region genes of kappa, lambda, alpha, gamma, delta, epsilon, and mu, as well as various immunoglobulin variable region genes. The light chain is classified as either kappa or lambda. The heavy chain is classified as gamma, mu, alpha, delta, or epsilon, which then define the classes of immunoglobulins IgG, IgM, IgA, IgD, and IgE, respectively. 【0025】

[0025] The basic structural unit of immunoglobulins (antibodies) is usually a tetramer. Each tetramer is composed of two identical pairs of polypeptide chains, each pair having one light chain (approximately 25 kD) and one heavy chain (approximately 50-70 kD). The N-terminus of each chain defines a variable region of approximately 100-110 or more amino acids, which is mainly involved in antigen recognition. Variable light chain (V L ) and variable heavy chain (V H The terms ) refer to these light and heavy chains, respectively. 【0026】

[0026] As used herein, the term antibody includes complete immunoglobulins and a plurality of distinct fragments or genetically engineered artificial antibodies produced by digestion with various peptidases. For example, pepsin digests an antibody under a disulfide bond in the hinge region to produce F(ab)'2, a dimer of Fab, which itself is V H -C H It is a light chain linked to 1. F(ab)'2 can be reduced under mild conditions to cleave the disulfide bond in the hinge region, thereby converting the F(ab)'2 dimer to a Fab' monomer. The Fab' monomer is substantially a Fab with a portion of the hinge region (see Fundamental Immunology, ed. WE Paul, Raven Press, NY, 1993). While various antibody fragments are defined in terms of digestion of a complete antibody, naturally, Fab' fragments may be newly synthesized either chemically or by utilizing recombinant DNA techniques. Thus, as used herein, the term antibody also includes antibody fragments that are produced by modification of a whole antibody or newly synthesized using recombinant DNA techniques. 【0027】

[0027] Antibodies for use in the methods, compositions, and systems of this disclosure may be monoclonal or polyclonal. For example, monoclonal antibodies can be prepared from mouse hybridomas according to the conventional method of Kohler and Milstein (Nature 256: pp. 495-497, 1975) or a method derived therefrom. Detailed procedures for the production of monoclonal antibodies are described in Harlow and Lane (Antibodies, A Laboratory Manual, CSHL, New York, 1988). 【0028】 【0028】The terms "specifically binds" and "specific binding" refer to the ability of a specific binding agent (such as an antibody) to bind to a target molecular species preferentially over binding to other molecular species when the specific binding agent and the target molecular species are mixed together with other molecular species. A specific binding agent is said to specifically recognize a target molecular species if it can specifically bind to its target. 【0029】 【0029】A single-chain antibody (scFv) is a genetically fused single-chain molecule linked by an appropriate polypeptide linker and contains one or more V H domains and V L domains of one or more antibodies (see, for example, Bird et al., Science, 242:423-426, 1988; Huston et al., Proc. Natl. Acad. Sci., 85:5879-5883, 1988). A diabody is a bivalent bispecific antibody in which the V H domains and V L domains are expressed on one polypeptide chain, but a linker that is too short is used to pair between the two domains of the same chain, thereby forcing these domains to pair with the complementary domains of another chain to create two antigen-binding sites (see, for example, Holliger et al., Proc. Natl. Acad. Sci., 90:6444-6448, 1993; Poljak et al., Structure, 2:1121-1123, 1994). One or more CDRs can be incorporated into the molecule either covalently or non-covalently, and the resulting molecule can be made into an immunoadhesin. The immunoadhesin may incorporate the CDR(s) as part of a larger polypeptide chain, covalently link the CDR(s) to another polypeptide chain, or incorporate the CDR(s) non-covalently. The CDR(s) enable the immunoadhesin to specifically bind to a particular antigen of interest. A chimeric antibody is an antibody that contains one or more regions of one antibody and one or more regions of one or more other antibodies. 【0030】

[0030] An antibody may have one or more binding sites. If there are more than one binding sites, they may be identical or different. For example, natural immunoglobulins have two identical binding sites, single-chain antibodies or Fab fragments have one binding site, while bispecific antibodies or bifunctional antibodies have two different binding sites. 【0031】

[0031] A neutralizing antibody or inhibitory antibody is an antibody that inhibits the activity of at least one of a target—usually a polypeptide—for example, by blocking the binding of that polypeptide to a ligand to which it normally binds, or by interrupting or otherwise interfering with the protein-protein interaction of the polypeptide with a second polypeptide. An activating antibody is an antibody that enhances the activity of a polypeptide. Antibodies can function as mimics of the activity of a target protein or as blockers of the activity of a target protein, from which a therapeutic effect is derived. 【0032】

[0032] Antigen: A compound, composition, or substance that can stimulate antibody production or a T cell response in an animal, and which includes compositions that are injected into or absorbed by the animal. Antigens include those that react with specific humoral or cellular immunity products and are induced by heterologous immunogens. 【0033】

[0033] Cavity: Any fluid-filled space inside a multicellular organism. In certain embodiments, a cavity may include several other cavities. For example, the pelvic cavity of a mammal includes the bladder, and the thoracic cavity includes cavities such as the upper digestive tract and the esophagus. In certain embodiments, a cavity may be the bladder and / or the renal pelvis and calyces and / or the urinary tract such as the ureters. 【0034】

[0034] Binding affinity: A term referring to the strength with which one molecule binds to another molecule at a site on that molecule. When a particular molecule binds to another particular molecule, or specifically binds to it, these two molecules are said to have binding affinity to each other. Binding affinity is related to the association constant and dissociation constant for a pair of molecules, but whether these constants are measured or determined is not important to the methods herein. Rather, the affinity used herein to describe the intermolecular interactions of the methods described is generally the apparent affinity (unless otherwise specified) observed in empirical studies, which can be used to compare the relative strength with which one molecule (e.g., an antibody or other specific binding partner) binds to two other molecules (e.g., two types or variants of a peptide). The concepts of binding affinity, association constant, and dissociation constant are well known. 【0035】

[0035] Cancer: A malignant disease characterized by the abnormal growth and differentiation of cells. The product of neoplasia is a neoplasm (tumor or cancer), which is the abnormal growth of tissue resulting from excessive cell division. Tumors that do not metastasize are called "benign." Tumors that invade surrounding tissues and / or can metastasize are called "malignant." Neoplasia is an example of a proliferative disorder. "Cancer cells" are cells that are neoplasms, for example, cells or cell lines isolated from a tumor. 【0036】

[0036] Solid tumors, such as sarcomas and carcinomas, and examples of solid tumors including cancers of the internal body cavity include fibrosarcoma, myxosarcoma, liposarcoma, chondrosarcoma, osteogenic sarcoma, and other sarcomas, synoviomas, mesothelioma, Ewing's tumor, leiomyosarcoma, rhabdomyosarcoma, colon cancer, lymphoid malignancies, pancreatic cancer, breast cancer, lung cancer (small cell lung cancer and non-small cell lung cancer, etc.), ovarian cancer, prostate cancer, hepatocellular carcinoma, squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, sweat gland carcinoma, medullary thyroid carcinoma, papillary thyroid carcinoma, and Examples include rom affinity cell carcinoma, sebaceous carcinoma, papillary carcinoma, papillary adenocarcinoma, medullary carcinoma, bronchogenic carcinoma, renal cell carcinoma, liver cancer, cholangiocarcinoma, choriocarcinoma, Wilms' tumor, cervical cancer, testicular cancer, seminomas, bladder cancer, melanoma, and CNS tumors (such as glioma, astrocytoma, medulloblastoma, craniopharyogioma, ependymoma, pinealoma, hemangioblastoma, acoustic neuroma, oligodendroglioma, meningioma, neuroblastoma, and retinoblastoma). 【0037】

[0037] Chemotherapy agent: An agent having therapeutic utility in the treatment of diseases characterized by abnormal cell growth or cell hyperplasia. Such diseases include cancer, autoimmune diseases, and diseases such as psoriasis characterized by hyperplastic growth. In one embodiment, the chemotherapeutic agent is a radioactive compound. Those skilled in the art can easily identify chemotherapeutic agents (see, for example, Slapak and Kufe, Chapter 86 of Principles of Cancer Therapy, Harrison's Principles of Internal Medicine, 14th edition; Perry et al., Chemotherapy, Chapter 17 of Abeloff, Clinical Oncology 2nd edition (copyright), 2000 Churchill Livingstone, Inc.; Baltzer L and Berkery R (eds.): Oncology Pocket Guide to chemotherapy, 2nd edition, St. Louis, Mosby-Year Book, 1995; Fischer DS, Knobf MF and Durivage HJ (eds.): The Cancer chemotherapy Handbook, 4th edition, St. Louis, Mosby-Year Book, 1993). Chemootherapeutic agents include small molecule agonists and biological agonists. Chemootherapeutic agents may be administered before, concurrently with, or after the described methods of cancer treatment using chemotherapy, immune checkpoint modulators, and thermoreversible hydrogel compositions. Exemplary chemotherapeutic agents include gemcitabine, mitomycin C, oxaliplatin, and docetaxel. 【0038】

[0038] Contact: Arranging to be directly, physically connected. This includes both solid and liquid forms. Contact can be induced in vitro using isolated cells or in vivo by administration to a subject. An exemplary form of contact is by dropper injection. 【0039】

[0039] Effective amount of compound: A sufficient amount of the compound to achieve the desired effect in the subject being treated. The effective amount of a compound, or "therapeutic effective amount," can be administered in single doses or multiple doses, for example, daily, for the duration of treatment. However, the effective amount of a compound will depend on the compound being applied, the subject being treated, the severity and type of the disease, and the method of administration of the compound. In particular examples, the effective amount of the activator in the described composition and treatment, i.e., at least one chemotherapy and at least one immune checkpoint modulator (i.e., inhibitor or inducer), is an amount that produces in the human subject receiving the composition a greater anticancer effect than that produced by treatment with either activator alone, and in particular embodiments, a synergistic anticancer effect. 【0040】

[0040] Efficacy: Refers to the ability of an agonist to produce a desired therapeutic effect. Efficacy also refers to the strength or potency of a compound. As used herein, "enhancement of efficacy" means increasing the therapeutic effect of an agonist. For example, if the agonist is a chemotherapeutic agent, "enhancement of efficacy" generally refers to increasing the ability of the agonist to kill target cells such as tumor cells. 【0041】

[0041] Increased risk: As used herein, “increased risk” of cancer refers to an increase in the statistical probability of developing or recurring cancer compared to the general population. For example, risk factors such as a history of bladder cancer may increase the risk of a subject experiencing a recurrence of bladder cancer. 【0042】

[0042] Intravesical instillation: A medical procedure that requires the direct / local administration of a drug into the bladder, also known as "intravesical therapy." Equivalent drug administration is possible for other body cavities. In certain embodiments, intravesical instillation requires drug delivery by catheter. In certain embodiments of the methods described herein, a hydrogel composition, such as a reversible (thermo-reversible) hydrogel, is administered to a subject by intravesical instillation without or without prior mixing with an active agent, such as, or not limited to, zarifremab, imiquimod, or gemcitabine. 【0043】

[0043] Inhibition of protein activity: Reducing, limiting, or blocking the action, function, or expression of a protein. The expression "inhibit protein activity" is not an absolute term. Rather, the expression conveys the broad inhibitory effect that various agonists may have on the normal (e.g., uninhibited or control) activity of a protein. Inhibition of protein activity may result in an increase in the level or activity of an indicator of protein activity, but is not necessarily required. For example, this increase may occur when the protein of interest acts as an inhibitor or suppressor of a downstream indicator. Thus, protein activity can be inhibited when the level or activity of any direct or indirect indicator of protein activity changes (e.g., increases or decreases) by at least 10%, at least 20%, at least 30%, at least 50%, at least 80%, at least 100%, or at least 250% or more compared to a control measurement of the same indicator. 【0044】

[0044] Immunotherapy: A method of inducing an immune response that counteracts the products of a target antigen. Immunotherapy based on cellular immune responses produces a cellular response against cells that produce a specific antigenic determinant, while immunotherapy based on humoral immune responses produces a specific antibody against a virus that produces a specific antigenic determinant. 【0045】

[0045] Pharmaceutically acceptable carriers: The activators for use in the methods described may be mixed with a pharmaceutically acceptable carrier. The pharmaceutically acceptable carriers useful in this disclosure are conventional. Remington's Pharmaceutical Sciences, by EW Martin, Mack Publishing Co., Easton, PA, 15th edition (1975), and its most recent edition, describe compositions and formulations suitable for the pharmaceutically acceptable delivery of the compounds disclosed herein. 【0046】

[0046] Pharmaceutical agent: A compound or composition that, when appropriately administered to a subject or cells, can induce a desired therapeutic or preventive effect. Incubation includes exposing the target to the activator for a sufficient time for the activator to interact with the cells. Contact includes incubating the activator in solid or liquid form with cells. 【0047】

[0047] Pharmaceutically acceptable carriers: The pharmaceutically acceptable carriers useful in this disclosure are conventional. Remington's Pharmaceutical Sciences, by E.W. Martin, Mack Publishing Co., Easton, PA, 15th edition (1975), and its most recent edition, describe compositions and formulations suitable for the pharmaceutically acceptable delivery of the compounds disclosed herein. 【0048】

[0048] Generally, the properties of the carrier will depend on the specific method of administration used. For example, parenteral formulations typically include injectable liquids containing a pharmaceutically and physiologically acceptable liquid as a vehicle, such as water, saline, equilibrium salt solution, dextrose aqueous solution, or glycerol. In the case of solid compositions (e.g., in the form of powders, pills, tablets, or capsules), conventional non-toxic solid carriers may include, for example, pharmaceutical-grade mannitol, lactose, starch, or magnesium stearate. In addition to a biologically neutral carrier, the pharmaceutical composition to be administered may contain small amounts of non-toxic auxiliary substances, such as wetting agents or emulsifiers, preservatives, and pH buffers, such as sodium acetate or sorbitan monolaurate. 【0049】

[0049] Prevention or treatment of disease: Prevention of disease refers to inhibiting the complete onset of a disease, for example, inhibiting the onset of myocardial infarction in a person with coronary artery disease, or inhibiting the progression or metastasis of a tumor in a subject with a neoplasm. Treatment refers to a therapeutic intervention that improves the signs or symptoms or pathological condition of a disease after it has started to develop. Prevention or treatment of disease may also refer to the results of an intervention taken to prevent recurrence of a disease that has previously received other treatment, such as surgery to remove a solid tumor in an internal body cavity. 【0050】

[0050] Radiation Therapy (Radiotherapy): Treatment of a disease (e.g., cancer or another disease or condition of overgrowth) by exposure of a subject or its tissue to a radioactive material. Radiation therapy is the medical use of ionizing radiation as part of cancer treatment to control malignant cells. Radiation therapy may be used for curative or adjunctive cancer treatment. Radiation therapy is used as palliative treatment when a cure is not possible and the aim is for local disease management or symptom relief. Radiation therapy may be administered before, concurrently with, or after described methods of cancer treatment using chemotherapy, immune checkpoint modulators and thermoreversible hydrogel compositions. 【0051】

[0051] Subjects susceptible to disease or condition: Subjects who can develop (or recur) a disease or condition, have a tendency to develop (or recur) a disease or condition, or have a predisposition to develop (or recur) a disease or condition. Naturally, subjects who already have or are exhibiting symptoms of a disease or condition are considered "suspicious" because they have already developed those symptoms. 【0052】

[0052] Therapeutic effective dose: A sufficient amount of the compound to achieve the desired effect in the subject being treated. The effective dose of the compound may be administered in single doses or multiple doses, for example, daily, during the treatment period. However, the effective dose will depend on the compound applied, the subject being treated, the severity and type of the disease, and the method of administration of the compound. For example, the therapeutic effective dose of the active ingredient can be measured as the concentration (moles per liter or molar M) of the active ingredient (such as a low molecular weight, peptide, protein, or antibody) in the blood (in vivo) or in a buffer (in vitro) that produces the effect. 【0053】

[0053] Thermoreversible Hydrogels: Thermoreversible hydrogels for use in the compositions and methods described are in liquid form at low temperatures and remain liquid during administration to a patient (e.g., by intravesical instillation). At high temperatures (e.g., typical human body temperature), thermoreversible hydrogels solidify, for example, in a covering of an internal body cavity. Low temperatures can be defined as less than 20°C, preferably less than 15°C. In certain embodiments, this low temperature can be less than 10°C. As used herein, thermoreversible hydrogels are synonymous with "reverse thermohydrogels" and "thermo-reversible hydrogels". Specific examples of thermoreversible hydrogels include compositions such as RTgel, which contain poloxamer with additional optional components. 【0054】

[0054] Urinary tract cancer: Cancer of any site in the urinary tract, including the urothelium, kidney, renal pelvis and ureter, bladder (also called "urinary bladder"), lamina propria, bladder muscle and urethra, as defined by T0, Ta, T1, T2, T3 and T4. Carcinoma in situ (CIS) is cancer found in the inner lining of the bladder endometrium. Intravesical instillation of chemotherapy after surgery is a common treatment for solid tumors of the urinary tract. Treatment using the described UGN-301 in combination with additional therapeutic agents such as TLR agonists and / or chemotherapeutic agents may also be provided for the treatment of cancers of the urinary tract as defined herein. 【0055】

[0055] Washing: Cleaning a part of a body using a liquid. In certain embodiments, “washing” a part of a body results in complete cleanliness of that part. In other embodiments, washing a part of a body does not result in complete cleanliness. In certain embodiments of the methods described, a body cavity is “washed” between administrations of a composition containing the hydrogel and the therapeutic agent. In certain embodiments, the composition is completely washed away from the body cavity. In other embodiments, the composition is not completely washed away from the body cavity. Washing (and removal of liquid) in the methods described herein can be achieved by standard methods of introducing liquid into a body cavity and removing it from the body cavity. 【0056】

[0056] Overview of several embodiments 【0057】

[0057] (i) Viscosity of 5000 cP or less at 5°C, and 1 × 10 at 17°C 6 cP~9×10 7 A pharmaceutical composition comprising a mixture of (ii) a thermoreversible hydrogel having a viscosity of cP and (ii) a liquid solution of zarefrelimab is described herein. 【0058】

[0058] In some embodiments, the pharmaceutical composition containing the thermoreversible hydrogel contains poloxamer, particularly 25% to 32% poloxamer. 【0059】

[0059] In some embodiments, the pharmaceutical composition containing a thermoreversible hydrogel contains a thickening agent. 【0060】

[0060] In another further embodiment, the pharmaceutical composition comprising a thermoreversible hydrogel comprises 20% to 32% (w / w) poloxamer, and the thickener is HPMC. 【0061】

[0061] In some embodiments, the pharmaceutical composition containing the thermoreversible hydrogel comprises 25% to 32% poloxamer, 0.02% to 0.5% HPMC, 0.75% to 1% PEG, and water. 【0062】

[0062] In some embodiments, the pharmaceutical composition comprises a liquid solution of zarefrelimab formulated in a solution containing histidine, sorbitol, methionine, and polysorbate 80, and is adjusted to a pH of 6-7. 【0063】

[0063] In some embodiments, the pharmaceutical composition comprises a liquid solution of zarefrelimab formulated in a solution containing 10-30 mM histidine, 200-300 mM sorbitol, 10-20 mM methionine, and 0.01-0.03% polysorbate 80, and is adjusted to a pH of 6-7. 【0064】

[0064] In some embodiments, the pharmaceutical composition contains zarifremab in a dose of 100 mg to 750 mg. 【0065】

[0065] In some embodiments, the pharmaceutical composition contains zarifremab at a concentration of 0.67 to 10 mg / mL. 【0066】

[0066] In another further embodiment, the pharmaceutical composition is formulated for intravesical administration into the bladder, and at least about 80% of the detectable zarifremab is released from the thermoreversible hydrogel within about 2 to about 12 hours. 【0067】

[0067] In other embodiments, the pharmaceutical composition further comprises at least one additional therapeutic agent, particularly mitomycin C, imiquimod, or gemcitabine. 【0068】

[0068] In some embodiments, the pharmaceutical composition supplies imiquimod at a concentration of about 1 mg / mL to about 10 mg / mL. 【0069】

[0069] In some embodiments, the pharmaceutical composition supplies gemcitabine in doses of about 100 mg to 3000 mg, particularly up to about 2000 mg. 【0070】

[0070] In some embodiments, the pharmaceutical composition has mitomycin C at a concentration of about 0.1 mg / mL to about 4 mg / mL. 【0071】

[0071] In some embodiments, after a single intravesical administration of the pharmaceutical composition, the urinary concentration of zarifremab exceeds 20 μg / mL. 【0072】

[0072] In some embodiments, when the pharmaceutical composition is stored at approximately 2°C to 8°C for at least 24 hours, it is substantially free of aggregates of zarefrelimab after 24 hours. 【0073】

[0073] In other embodiments, the thermoreversible hydrogel further comprises at least a stabilizer, an active oxygen scavenger, a surfactant, a buffer, and a gelation temperature regulator. 【0074】

[0074] In some embodiments, the thermoreversible hydrogel has a gelation temperature of about 10°C to about 20°C, particularly 13°C to about 16°C. 【0075】

[0075] In other embodiments, the poloxamer is poloxamer 407. 【0076】

[0076] In certain embodiments, the pH of the composition is approximately 6 to approximately 8. 【0077】

[0077] Further described herein are pharmaceutical compositions for use in the treatment of bladder cancer, wherein a thermoreversible hydrogel is administered intravesically into the bladder of a subject, thereby resulting in a urinary concentration of at least 20-70 μg / mL of zarifremab in the urine of the subject at least 2 hours after administration. 【0078】

[0078] Further described herein are kits for preparing a pharmaceutical composition, comprising (i) a thermoreversible hydrogel containing 20% ​​to 32% poloxamer 407, and (ii) a solution containing 100 mg to 750 mg of zarifremab, wherein mixing (i) and (ii) forms about 20 to 100 mL of the pharmaceutical composition. 【0079】

[0079] The Specified Specified Kit for Preparing a Pharmaceutical Composition also comprises (i) 60 to 73 mL of a thermoreversible hydrogel containing 20% ​​to 32% poloxamer 407, and (ii) 2 to 15 mL of a solution containing 100 mg to 750 mg of zarifremab, wherein mixing (i) and (ii) forms approximately 75 mL of the pharmaceutical composition. 【0080】

[0080] Further described herein is a kit for preparing a pharmaceutical composition, comprising (i) about 40 to 80 mL of a thermoreversible hydrogel, (ii) about 1 to 10 mL of a mixture of a stabilizer, an active oxygen scavenger, a surfactant, a buffer, and a gelling temperature regulator, and (iii) about 10 to 20 mL of a solution containing 50 mg to 750 mg of zarifremab, wherein mixing (i), (ii), and (iii) forms about 50 to 100 mL of the pharmaceutical composition. 【0081】

[0081] Ready-to-use pharmaceutical compositions for administration to a patient, comprising zarefrelimab, poloxamer, stabilizers, buffers, reactive oxygen species scavengers, and gelling temperature regulators, are also described herein. 【0082】

[0082] In some embodiments, the pharmaceutical composition contains a thermoreversible hydrogel comprising 25% to 32% poloxamer, 0.02% to 0.5% HPMC, and water. 【0083】

[0083] In certain embodiments, the bladder cancer is high-grade or low-grade NMIBC, MIBC, or carcinoma in situ (CIS). 【0084】

[0084] Thermoreversible hydrogel and its properties 【0085】

[0085] A pharmaceutical composition for sustained local delivery comprising a mixture of an immunomodulatory antibody and a thermoreversible hydrogel for administration into a body cavity for the treatment of cancer such as bladder cancer is described herein. In certain embodiments, the pharmaceutical composition of the Disclosure is a thermoreversible hydrogel that is mixed with a stable solution of an activator before administration to a patient. Such a hydrogel has a relatively low viscosity at low temperatures but at higher temperatures (e.g., body temperature) forms a semi-solid gel that can act as a drug depot in a body cavity such as the bladder, providing sustained release of an activator such as zarifremab over a period of about 3 to about 24 hours, including 3 hours, 4 hours, about 5 hours, about 6 hours, about 7 hours, about 8 hours, about 9 hours, about 10 hours, about 11 hours, or about 12 or about 14 or about 16 or about 18 or about 20 or about 24 hours (including all ranges between any of these values). The sustained release of active agents such as zarifremab provided by the compositions of this disclosure results in improved efficacy and tolerability compared to other types of compositions and administration methods, such as systemic administration. The semi-solid gel formed after administration of the compositions of this disclosure into the patient's body cavity slowly disintegrates over time and, when administered intravesically to the bladder, is excreted by normal urinary flow. 【0086】

[0086] The thermoreversible hydrogel of the present invention comprising at least one grade of poloxamer. Poloxamer is a nonionic triblock copolymer comprising a central chain of relatively hydrophobic polypropylene oxide (PO) flanked on both sides by two relatively hydrophilic polyethylene oxide (EO) chains. The PO and EO chains may be of various lengths and can be expressed by the average number of polypropylene oxide monomer units and ethylene oxide monomer units. Various grades of poloxamers, such as those described in U.S. Patent No. 9,040,074 and U.S. Patent No. 9,950,069, are suitable for use in the thermoreversible hydrogel compositions of the present disclosure, each of which is incorporated herein by reference in whole for all purposes. Suitable poloxamers include those manufactured by BASF under the trademark KOLLIPHOR, such as KOLLIPHOR 407. 【0087】

[0087] In certain embodiments, the thermoreversible hydrogels of the Disclosure contain a poloxamer such as poloxamer 407 in an amount of about 20 wt.% to about 32 wt.%, and include about 22 wt.%, about 24 wt.%, about 25%, about 28 wt.%, about 29 wt.%, about 30 wt.%, or about 31 wt.% or about 32% (including any range between any of these values) of KOLLIPHOR 407 (or similar poloxamers having substantially the same composition). Further examples of the claimed thermoreversible hydrogels are described in U.S. Patent No. 10,758,482, which is incorporated herein by reference in its entirety. 【0088】

[0088] The compositions of the present disclosure may also be characterized by their heat-dependent properties (e.g., temperature-dependent viscosity and / or gelation temperature). The compositions of the present disclosure have a viscosity of about 1000 cP or less at 5°C (e.g., viscosities of less than about 100 cP, about 100 cP, about 200 cP, about 300 cP, about 400 cP, and a viscosity range of about 100 cP, about 200 cP, about 300 cP, or about 400 cP or about 500 cP, or any value between about 600 cP, about 700 cP, about 800 cP, about 1000 cP, all measured at 5°C). The compositions of the present disclosure have a viscosity of about 1 × 10 at 19°C. 6 cP~approx. 1.5×10 7 It has a viscosity in the range of cP, approximately 2 × 10⁻⁶ 6 cP, approx. 5×10 6 cP, approx. 6×10 6 cP, approx. 7×10 6 cP, approx. 8×10 6 cP, approx. 9×10 6 cP, approx. 1.0×10 7 cP, approx. 1.1×10 7 cP, approx. 1.2×10 7 cP, approx. 1.3×10 7 cP, approx. 1.4×10 7 cP, or approximately 1.5 × 10⁻⁶ 7 This includes cP and encompasses the entire range between any of these values ​​(all measured between 17 and 21°C, especially at 19°C). 【0089】

[0089] The viscosities described herein are measured using a viscometer that is common in the art, such as a Brookfield viscometer, but not limited to this. 【0090】

[0090] The gelation temperature of the hydrogels of the Disclosure defines the temperature at which the compositions of the Disclosure exhibit a significant increase in viscosity. The thermoreversible hydrogels of the Disclosure have a gelation temperature in the range of about 10°C to about 20°C, including about 10°C, about 11°C, about 12°C, about 13°C, about 14°C, about 15°C, about 16°C, about 17°C, about 18°C, about 19°C, and about 20°C, and encompass all ranges between any of these values. In certain embodiments, the gelation temperature is in the range of about 11°C to about 20°C, or about 12°C to about 14°C. 【0091】

[0091] In certain embodiments, the pharmaceutical compositions of the present disclosure include zarefrelimab and one or more of the following: stabilizers, reactive oxygen species scavengers, surfactants, buffers, thermoreversible hydrogels, gelation temperature regulators, and thickeners. 【0092】

[0092] In certain embodiments, examples of thickeners include modified cellulose, such as hydroxypropyl methylcellulose, methylcellulose, hydroxypropylcellulose, carboxymethylcellulose and their salts, ethylcellulose, etc.; synthetic polymers, such as polyacrylic acid and polymethacrylate, etc.; gelatin, starch and starch derivatives, gums, such as guar gum and xanthan gum, etc.; and any combination of the aforementioned thickeners. In certain embodiments, the compositions of the present disclosure include hydroxypropyl methylcellulose (HPMC). 【0093】

[0093] Suitable stabilizers for the compositions of the present disclosure include sugars or sugar alcohols, such as sucrose, lactose, glucose, fructose, maltose, mannitol, sorbitol, xylitol, lactitol, and combinations thereof. In certain embodiments, the compositions of the present disclosure include sorbitol. 【0094】

[0094] Methionine is a suitable reactive oxygen species scavenger for the compositions of the present disclosure. In certain embodiments, the compositions of the present disclosure include methionine. 【0095】

[0095] Suitable surfactants for the compositions of the present disclosure include polysorbates (e.g., polysorbate 80), sodium dodecyl sulfate, dextran sulfate, and combinations thereof. In certain embodiments, the compositions of the present disclosure include polysorbate 80. 【0096】

[0096] Suitable buffers for the compositions of the present disclosure include buffers that achieve a pH in the range of about 6 to about 8. In some embodiments, suitable buffers achieve a pH in the range of about 6 to about 8 at a temperature of about 2 to 8°C. Specific buffers include histidine buffers and phosphate buffers. In certain embodiments, the compositions of the present disclosure include a histidine buffer. 【0097】

[0097] Examples of the thermoreversible hydrogel compositions described include triblock copolymers having the general form of ABA copolymer or BAB copolymer, in which case A is a hydrophilic block and B is a hydrophobic block. More specifically, A or B is selected from PEO((poly(ethylene oxide))), PLGA(poly(lactic acid-co-glycol) acid, PLA(polylactic acid) and PPO(polypropylene oxide), PGA(polyglycolic acid), PCL-(polycaprolactone), PCLA-poly(ε-caprolactone-co-lactide), PCBCL-poly(α-carboxylate-co-α-benzylcarboxylate-ε-caprolactone). Selected or alternatives include glycolide, lactide, e-caprolactone, p-dioxanone (l,4-dioxan-2-one), trimethylene carbonate (l,3-dioxan-2-one), l,4-dioxepant-2-one (including its dimer l,5,8,l2-tetraoxacyclotetradecane-7,l4-dione), l,5-dioxepant-2-one, 6,6-dimethyl-l,4-dioxan-2-one, and 2,5-diketomol. Foline, pivalolactone, kai-diethylpropiolactone, ethylene carbonate, ethylene oxalate, 3-methyl-l,4-dioxan-2,5-dione, 3,3-diethyl-l,4-dioxan-2,5-dione, 6,8-dioxabicycloctan-7-one, beta-propiolactone, gama-butyrolactone, delta-valerolactone, epsilon-decalactone, 3-methyl-l,4-dioxan-2,5-dione, l,4-di It comprises at least two cyclic monomers selected from the group consisting of oxan-2,5-dione, 2,5-diketomorpholine, alpha,alpha-diethylpropiolactone, gama-butyrolactone, l,4-dioxepant-2-one, l,5-dioxepant-2-one, 6,6-dimethyl-dioxepant-2-one, 6,6-dioxabicycloctan-7-one, or 5,5-dimethyl-l,3-dioxan-2-one. 【0098】

[0098] In other embodiments, the thermoreversible hydrogel composition comprises at least one triblock copolymer having the general form of an ABA copolymer or a BAB copolymer, where A is a hydrophilic block and B is a hydrophobic block. In specific embodiments, the triblock copolymer has the ABA form of an EPO / PPO / EPO block copolymer. 【0099】

[0099] Suitable gelling temperature regulators include polyethylene glycol (PEG), such as PEG200, PEG300, PEG400 and PEG-800. In certain embodiments, the compositions of the present disclosure include PEG-400. 【0100】

[0100] In other embodiments of the present disclosure, the poloxamer includes poloxamer 407, poloxamer 188, poloxamer 124, poloxamer 237, or poloxamer 338 or a combination thereof, having an average molecular weight of 8500 to 16000 DA (including upper and lower limits). 【0101】

[0101] In certain embodiments, the thermoreversible hydrogel of the Disclosure comprises 24-32% poloxamer 407, 0.04-2% HPMC, and 0.5-2% PEG400 and water. In certain embodiments, the hydrogel of the Disclosure comprises 31% poloxamer 407, 0.05% HPMC, 1% PEG400 and water. 【0102】

[0102] In certain embodiments, the poloxamer component in the pharmaceutical composition of the present invention is present at a concentration of 20% to 30% (w / w) and any range within that, for example, 22% to 30%, 23% to 28%, or 24% to 26% of the pharmaceutical composition. In another particular embodiment, the poloxamer is present at a concentration of 24.8% of the pharmaceutical composition. 【0103】

[0103] In certain embodiments, the thermoreversible hydrogel composition may further include at least one of the following: a thickening agent, a mucosal adhesion enhancer, an elution rate controller, a gelation temperature controller, a pH controller, a thickening agent, a stabilizer, an active oxygen scavenger, a surfactant, a buffer, and an additional gelation temperature regulator, an absorption enhancer / tight junction regulator / cell membrane permeability enhancer, an organic acid, or a cyclodextrin. 【0104】

[0104] In certain embodiments, the dissolution rate control agent may include, but is not limited to, silicon dioxide or any derivative thereof, nanoparticles or microparticles of poly(lactide-co-glycolide (PLGA), polylactic acid (PLA), polyglycolic acid (PGA), PLA-PEG copolymer or PLGA-PEG copolymer, nanoparticles or microparticles of polystyrene or polymethyl methacrylate (PMMA), calcium carbonate, microcrystalline cellulose, aluminum hydroxide, Eudragit.RTM.NE, Eudragit.RTM.RS and RL, cellulose acetate and cellulose acetate butyrate, crospovidone, crosslinked carboxymethylcellulose sodium, crosslinked starch glycolate sodium, thickeners, soy, iodide aromatic compounds, cyclodextrin, and cholesterol. 【0105】

[0105] In certain embodiments, gelation temperature control agents include, but are not limited to, urea, polyethylene glycol, short-chain fatty acids and their salts (sodium octanoate, sodium dodecyl sulfate), ethanol, glyceryl monostreatrate, isopropyl myristate, and polysorbate surfactants. 【0106】

[0106] In certain embodiments, the pharmaceutical composition can be sterilized. 【0107】

[0107] In various embodiments, the composition of the present invention comprises about 100 mg to about 900 mg of zarifremab, for example, about 100 mg, about 150 mg, about 200 mg, about 250 mg, about 300 mg, about 350 mg, about 400 mg, about 450 mg, about 500 mg, about 550 mg, about 600 mg, about 650 mg, or about 700 mg, about 750 mg, about 800 mg, about 850 mg, or about 900 mg of zarifremab, encompassing all ranges between any of these values. 【0108】

[0108] In various embodiments, the concentration of zarifremab in the pharmaceutical composition of the Disclosure is in the range of about 1 mg / mL to about 11 mg / mL, and includes any of the following values ​​up to about 1 mg / mL, about 1.3 mg / mL, about 2 mg / mL, about 2.7 mg / mL, about 3.3 mg / mL, about 4 mg / mL, about 4.7 mg / mL, about 5.3 mg / mL, about 6 mg / mL, about 6.7 mg / mL, about 7 mg / mL, about 7.9 mg / mL, about 8.6 mg / mL, about 9.3 mg / mL, about 10 mg / mL, about 10.5 mg / mL, or about 11 mg / mL, 50 mg / mL, or 60 mg / mL, and includes all ranges between any of these values. 【0109】

[0109] In certain embodiments, the composition of the present disclosure comprises about 100 mg to about 750 mg of zarifremab, poloxamer, a histidine buffer to achieve a pH in the range of about 6 to 8, hydroxypropyl methylcellulose, PEG-400, methionine, sorbitol, polysorbate 80, and water, thereby forming a thermoreversible hydrogel. 【0110】

[0110] In certain embodiments, the disclosure relates to a pharmaceutical composition in the form of a thermoreversible hydrogel comprising poloxamer, water, and about 100 mg to about 750 mg of zarifremab. Such a composition has a viscosity of 1000 cP or less at 5°C and 1 × 10⁻¹⁶ at 19°C. 6 cP ~ 1.5 × 10 7 It has a viscosity of cP. 【0111】

[0111] In another further specific embodiment, the pharmaceutical composition comprises poloxamer, water, about 100 mg to about 750 mg of zariflemab, a thickener, a stabilizer, an active oxygen scavenger, a surfactant, a buffer, a solvent / cosolvent, and optionally a gelation temperature regulator. In a specific embodiment, the thickener is hydroxypropyl methylcellulose, the stabilizer is sorbitol, the active oxygen scavenger is methionine, the surfactant is polysorbate 80, and the buffer is a histidine buffer. 【0112】

[0112] Pharmaceutical composition and method of treatment 【0113】

[0113] Pharmaceutical compositions comprising a reverse-thermal hydrogel combined with an activator, such as an antibody, for example, a monoclonal antibody, or for example, an immune checkpoint inhibitor (for example, an antibody or fragment thereof that specifically binds to CTLA-4, PDL-1, or PD-1 and inhibits their activity), are described herein. In other embodiments, the activator may be an antibody-drug conjugate (ADC), which is a product in which a monoclonal antibody is linked to a small molecule drug by a stable linker, for example, gemtuzumab-ozogamicin. In certain embodiments, the immune checkpoint inhibitor may be an inhibitor that is anti-PD1, anti-PDL1, anti-CTLA4, anti-KIR, anti-LAG3, anti-VISTA, anti-TIM3, anti-B7-H3, anti-B7-H4, and anti-BTLA. In certain embodiments, the immune checkpoint modulator is an antagonist antibody, such as pembrolizumab (Keytruda, MK-3475), semiprimab (AGEN-2034), nivolumab (Opdivo, BMS-936558), pizilizumab (CT-011), MEDI0680 (AMP514), TSR-042, AMP-224, durvalumab (MEDI4736), avelumab (MSB0010718C), atezolizumab (MPDL3280A), CK-301, MDX1105 (BMS-936559), ipilimumab (Yervoy), tremelimumab (CP-675, 206), These are lirulumab (BMS986015), IPH2101, relatrimab (BMS986016), IMP321, TSR-033, JNJ-61610588, CA-170, BMS-986207, TSR022, enobrituzumab (MGA271), MGD009, zarifrerimab (AGEN-1884), AGEN-2041 (AGEN-1181), ADU-1604, AK104, ALPN-202, BCD-145, BMS-986249, BPI-002, CBT-509, ADU-1604, ATOR1144, ATOR-1015, and DS-5573a. The representative immune checkpoint inhibitor used in this specification is zarifremab (AGEN-1884). 【0114】

[0114] The pharmaceutical compositions described are used in methods for treating cancer and as compositions for use in treating cancer. In certain embodiments, cancer is urinary tract cancer, kidney cancer, or bladder cancer. The pharmaceutical compositions of the present invention can be administered either before or after standard tumor resection surgery. In some embodiments, the compositions described inhibit tumor regrowth after surgery. In other embodiments, the compositions described reduce tumor size before surgery. In yet another embodiment, the compositions described reduce or even eliminate the presence of a tumor in the bladder. 【0115】

[0115] In certain embodiments, the composition described comprises a reverse thermal hydrogel mixed with a zarifremab solution. Such a composition is referred to herein as UGN-301. 【0116】

[0116] Zarifremab is a recombinant fully human immunoglobulin G1 monoclonal antibody that neutralizes the inhibitory effect of CTLA-4 on the immune response of tumor-specific T cells. Zarifremab aims to enhance tumor immune surveillance and anti-tumor immune responses by blocking CTLA-4 from binding to its ligands (cluster of differentiation [CD]80 and CD86), thereby neutralizing the negative regulatory effect of CTLA-4 on T cells. 【0117】

[0117] Zarifremab was originally identified from a human donor-derived cell antibody library, and an intravenous (IV) formulation is currently under development as monotherapy in patients with advanced or refractory cancer, and as combination therapy with valstirimab (anti-PD1 antibody) in patients with metastatic or locally advanced solid tumors (including cervical cancer, inoperable or recurrent angiosarcoma, and metastatic, locally advanced, and / or unresectable squamous cell carcinoma, adenosquamous carcinoma, or cervical adenocarcinoma). 【0118】

[0118] In yet another embodiment, the described method provides an immune checkpoint inhibitor, in particular zarifremab, in combination with an additional active chemotherapeutic agent or other anticancer agent (such as but not limited to mitomycin C (MMC), imiquimod (UGN201), and gemcitabine), or any pharmaceutical composition comprising one of mitomycin C (MMC), imiquimod, and gemcitabine, mixed with a thermoreversible hydrogel. 【0119】

[0119] As further described herein, additional activators can be formulated in a thermoreversible hydrogel composition together with a checkpoint inhibitor such as zarifremab, or separately in a solution formulation suitable for intravesical administration. Prior to combination with the hydrogel, such activators can be supplied in various forms, including stable solutions, solid forms, or lyophilized forms. In certain embodiments, the additional activator is imiquimod, or gemcitabine or MMC, referred herein as UGN-201. UGN-201 is a formulation of imiquimod, a Toll-like receptor (TLR7) agonist, optimized for intravesical administration into the bladder. This route of administration allows for direct exposure to target tissue and potentially provides effective treatment of NMIBC while minimizing systemic exposure to imiquimod or small molecule chemotherapeutic agents (such as, but not limited to, mitomycin or gemcitabine). 【0120】

[0120] In other embodiments, UGN-301 is provided as a combination treatment with UGN-201 (imiquimod), or an additional active ingredient such as MMC or gemcitabine, or other chemotherapeutic agents, not limited to these. The additional agent is administered once a week for 6 to 12 weeks at the same frequency as UGN-301. In certain embodiments, the additional agent is administered at a lesser or more frequent frequency than UGN-301, as determined by a skilled clinician. 【0121】

[0121] In certain embodiments, UGN-201, a TLR-7 agonist, is administered at a concentration of 1 mg / ml to 10 mg / ml [50 mg to 500 mg] before, after, or simultaneously with the administration of UGN-301. In other embodiments, UGN-201 is administered at a dose of 200 mg [at a concentration of 4 mg / ml]. In some embodiments, 200 mg of UGN-201 is administered at a concentration of 2 to 8 mg / mL. 【0122】

[0122] There may be an optional wash between the administration of UGN-201 and the administration of UGN-301. The wash may last for 0-10 minutes, 10-20 minutes, 20-30 minutes, 30-40 minutes, 40-50 minutes, 50-60 minutes, or up to 2-3 hours, or any time frame in between. 【0123】

[0123] In additional embodiments, UGN-201 may remain in the bladder or bladder cavity alone for up to one hour, up to two hours, up to three hours, or any time in between. 【0124】

[0124] As described herein, pharmaceutical compositions of the present invention, such as UGN301, designed for local intravesical administration, prolong the release of zarifremab in the bladder and increase the concentration of localized zarifremab, while avoiding significant systemic exposure due to immune activation associated with CTLA-4 blockade and reducing systemic toxicity. 【0125】

[0125] The compositions of the present disclosure may be provided for use in the treatment of patients with urinary tract cancer and bladder cancer, such as muscle-non-muscle-invasive bladder cancer (MIBC), muscle-invasive bladder cancer (MIBC), and metastatic bladder cancer, including moderate-risk, low-grade, high-grade, papillary (Ta), lamina propria-invasive (T1), and carcinoma in situ (CIS). For example, the disclosed pharmaceutical compositions may be formulated as a thermoreversible hydrogel containing about 100 mg to about 750 mg of zarifremab, or alternatively, as a kit comprising (a) an aqueous solution of 100 mg to 750 mg of zarifremab and (b) a thermoreversible hydrogel. In the case of the kit, the aqueous solution of zarifremab (a) is immediately mixed with the thermoreversible hydrogel (b) within 12, 24, 48, 72, 96 hours, or 28 hours prior to administration. This composition or kit is considered a monotherapy treatment, and the sole active ingredient in the composition is an anti-CTLA-4 antibody such as zarifremab. The composition can be administered to the patient once a week in a 6-week course, followed by an optional maintenance period, during which the patient receives up to 6 additional doses of the composition between 6 and 12 months after the initial dose. 【0126】

[0126] In certain embodiments, the zarifrelimab solution contains about 100 mg to about 750 mg of zarifrelimab in a volume of about 15 mL, and the thermoreversible hydrogel (as described herein) has a volume of about 60 mL. In certain examples, the therapeutic composition obtained after mixing the zarifrelimab solution and the thermoreversible hydrogel is a hydrogel of about 75 mL containing about 100 mg to about 750 mg of zarifrelimab, although the volume of the combined solution containing zarifrelimab together with the hydrogel composition can vary from 30 mL to 150 mL. 【0127】

[0127] The composition of the zarifremab solution can be any aqueous composition suitable for preventing the loss of activity of zarifremab for about 60 months when stored at about 2 to 8°C. 【0128】

[0128] In certain embodiments, the zarifremab solution is formulated by combining approximately 30–100 mg / mL of zarifremab with 10–30 mM histidine, 200–300 mM sorbitol, 10–20 mM methionine, and 0.01–0.03% polysorbate 80, and is stable for up to 60 months when stored at 2–8°C and adjusted to a pH of 6–7 according to a standard method. In certain embodiments, the zarifremab solution for combination with a poloxamer-based thermoreversible hydrogel contains 20 mM histidine, 250 mM sorbitol, 15 mM methionine, and 0.02% polysorbate 80. In such and additional embodiments, the hydrogel composition may include 25% to 31% poloxamer, 1% PEG, and 0.05% to 2% HPMC, along with 100 to 700 mg of zarifremab used. In optional embodiments, the same diluent as zarifremab (20 mM histidine, 250 mM sorbitol, 15 mM methionine, and 0.02% polysorbate 80) is also used to dilute the concentration of the thermoreversible hydrogel. 【0129】

[0129] In various other embodiments, the relative volume of the zarefrelimab solution (multiple types are possible) and the thermoreversible hydrogel may differ, however, when mixed together, the amount of zarefrelimab in the resulting thermoreversible hydrogel is in the range of approximately 100 mg to approximately 750 mg, the total volume of the thermoreversible hydrogel containing zarefrelimab is in the range of approximately 50 mL to 100 mL, and the viscosity characteristics and gelation temperature characteristics of the composition are consistent with those described herein. For example, the volume of the zarifremab solution(s) (multiple types are possible) can be in the range of approximately 10 mL to approximately 20 mL (including approximately 10 mL, approximately 15 mL, or approximately 20 mL, and including any range between these values), and the volume of the thermoreversible hydrogel can be in the range of approximately 40 mL to approximately 100 mL (including approximately 40 mL, approximately 45 mL, approximately 50 mL, approximately 55 mL, approximately 60 mL, approximately 65 mL, or approximately 70 mL, approximately 80 mL, approximately 90 mL, or approximately 100 mL, and including any range between these values). 【0130】

[0130] The compositions of this disclosure are administered intravesically to the bladder of patients with bladder cancer, including high-grade nonmuscle-invasive bladder cancer, such as papillary (Ta), lamina propria invasive (T1), and carcinoma in situ (CIS), muscle-invasive bladder cancer (MIBC), and metastatic bladder cancer. Typical administration volumes range from about 50 mL to about 100 mL, depending on the volume required by the particular patient. In most embodiments, a single dose of the composition described herein (e.g., about 50 mL, 60 mL, 70 mL, 75 or 80 mL of the thermoreversible hydrogel composition described herein containing about 100 mg to about 750 mg of zarifremab) is administered intravesically once a week for at least about 4 to 6 weeks. Thereafter, single or multiple doses of the disclosed pharmaceutical composition may be administered every 12 weeks. In another embodiment, intravesical administration of the composition described may be once weekly for up to 12 weeks, particularly once weekly for up to 6 weeks, once every 3 weeks for 12 weeks, once every 2 weeks for 12 weeks, or twice weekly for 6 weeks. Alternatively, administration of the composition may be once monthly for 6, 9, and 12 months after the initial administration, or up to three doses every 12 weeks over a longer period, or three additional doses over 6 to 12 months after the initial administration, or additional doses as determined by an experienced clinician. 【0131】

[0131] In another further embodiment of the present disclosure, intravesical administration of the composition described is used to treat bladder cancer once a week for up to four times prior to cystectomy. 【0132】

[0132] In some embodiments, patients with bladder cancer are treated with a composition of the present invention containing zarifrelimab in combination with a therapeutic dose of one or more additional active agents, such as chemotherapeutic agents or TLR agonists, used or developed for the treatment of bladder cancer, as described herein. This treatment is considered a combination treatment comprising zarifrelimab plus at least one additional active ingredient, such as, but not limited to, a TLR7 agonist such as imiquimod, or a chemotherapeutic agent such as gemcitabine or mitomycin C. In some embodiments, the additional active agent is administered before or after the composition containing zarifrelimab. The additional active agent may be formulated in the same thermoreversible hydrogel containing zarifrelimab, so that zarifrelimab and the additional active agent (e.g., imiquimod, mitomycin, or gemcitabine) are administered together or sequentially into the bladder, with optional rinsing between the two components. Alternatively, the thermoreversible hydrogel of this disclosure containing zarifrelimab and an additional activator (e.g., imiquimod) are administered separately (both intravesically). When administered separately, the thermoreversible hydrogel of this disclosure containing zarifrelimab may be administered either before or after the additional activator (e.g., imiquimod, mitomycin, or gemcitabine). The interval between the administration of the thermoreversible hydrogel containing zarifrelimab and the administration of the additional activator may be at least 1 hour to about 36 hours, or 48 hours to 72 hours, with optional lavage between the administration of zarifrelimab and the administration of the additional activator. In certain embodiments of this disclosure, mitomycin is administered in amounts of 20 to 200 mg. In another further embodiment of this disclosure, gemcitabine is administered in a gel formulation in amounts of 100 to 1000 mg, or particularly 50 to 500 mg. In other embodiments, gemcitabine is administered by dropwise infusion in a liquid formulation in an amount between 500 and 2000 mg. 【0133】

[0133] When administered in combination with the thermoreversible hydrogel of this disclosure containing zarifremab, additional activators (e.g., imiquimod, mitomycin, or gemcitabine, but not limited to those listed below) may be administered in any dosage form, preferably intravesically. The dosage regimens of the various combinations described herein (e.g., zarifremab and additional activators), and the provision of activators, vary depending on the activator. In all regimens, zarifremab is provided in a reverse-heat gel as described. Additional activators (multiple types) may be provided beforehand, with or without washing in between, together with the antibody in the same reverse-heat gel formulation as the antibody, and / or after antibody treatment. In certain embodiments, the activator provided in addition to the checkpoint inhibitor antibody is provided in an intravesical infusion solution. In other embodiments, the additional activator is provided in a reverse-heat gel solution to allow for prolonged contact between the additional activator and the target site. 【0134】

[0134] The compositions of the present disclosure are effective in treating low-grade and high-grade bladder cancers, such as non-muscle-invasive bladder cancer (NMIBC), particularly BCG-unresponsive NMIBC, and bladder cancers including MIBC. Drugs such as zarifrimab are effective immunotherapies, but when administered systemically, they may have toxic side effects that limit both the dose and the schedule. The disclosed intravesical thermoreversible hydrogel avoids systemic administration of zarifrimab because zarifrimab is applied locally / topically to the target site, for example, to the site of bladder cancer. However, drugs applied locally / topically are not always therapeutically effective because the drug must be released from the local / topical composition at a certain concentration and contact time, and diffuse through the patient's tissues to deliver a therapeutically effective dose to the target site. Therapeutic antibodies such as zarifremab must also be sufficiently stable in the therapeutic formulation to maintain therapeutic levels of activity. Therefore, while there are examples in the art of therapeutically effective topically applied drug formulations, those skilled in the art recognize that the therapeutic efficacy of topically / topically applied antibodies such as zarifremab is unpredictable. As described herein, a thermoreversible hydrogel administered intravesically, containing approximately 100 mg to approximately 750 mg of zarifremab in combination with at least one additional activator of optional choice, is effective in the treatment of bladder cancer as described herein. 【0135】

[0135] The thermoreversible hydrogels of the present disclosure, when administered intravesically into a patient's bladder, are effective in achieving and maintaining therapeutically effective levels of local / topical contact of zarifremab in the bladder for at least 4 hours post-administration. In some embodiments, post-administration local exposure to zarifremab is maintained for about 4 to about 24 hours post-administration, including about 4 hours, about 5 hours, about 6 hours, about 7 hours, about 8 hours, about 9 hours, about 10 hours, about 11 hours, or about 12 hours, and including all ranges between any of these values. 【0136】

[0136] In various embodiments, after a single intravesical administration of approximately 70 mL of thermoreversible hydrogel containing 100 mg of zarifremab, the range of the area under the pharmacokinetic curve (AUC) of zarifremab in urine is approximately 907 to 1194 h*ug / mL. 【0137】

[0137] In various embodiments, after a single intravesical administration of approximately 70 mL of thermoreversible hydrogel containing 300 mg of zarifremab, the AUC of zarifremab in urine is in the range of approximately 2330 to 7400 hr*ug / mL. 【0138】

[0138] In various embodiments, after a single intravesical administration of approximately 70 mL of thermoreversible hydrogel containing 500 mg of zarifremab, the AUC of zarifremab in urine is in the range of approximately 3708 to 7758 hr*ug / mL. 【0139】

[0139] In various embodiments, after a single intravesical administration of approximately 70 mL of thermoreversible hydrogel containing 700 mg of zarifremab, the AUC of zarifremab in urine is in the range of approximately 4400 to 10000 hr*ug / mL. 【0140】

[0140] In various embodiments, after a single intravesical administration of approximately 70 mL of thermoreversible hydrogel containing 100 mg of zarifremab, the range of urinary zarifremab Cmax is approximately 130 to 272 ug / mL. 【0141】

[0141] In various embodiments, after a single intravesical administration of approximately 70 mL of thermoreversible hydrogel containing 300 mg of zarifremab, the Cmax of zarifremab in urine is in the range of approximately 506 to 897 ug / mL. 【0142】

[0142] In various embodiments, after a single intravesical administration of approximately 70 mL of thermoreversible hydrogel containing 500 mg of zarifremab, the Cmax of zarifremab in urine is in the range of approximately 628 to 1604 ug / mL. 【0143】

[0143] In various embodiments, after a single intravesical administration of approximately 70 mL of thermoreversible hydrogel containing 700 mg of zarifremab, the Cmax of zarifremab in urine is in the range of approximately 500 to 2000 ug / mL. 【0144】

[0144] In various embodiments, after a single intravesical administration of approximately 70 mL of thermoreversible hydrogel containing 100 mg of zarifremab, the range of urinary zarifremab AUC is approximately 0.75 to 1.5 mg*hr / mL. 【0145】

[0145] In various embodiments, after a single intravesical administration of approximately 70 mL of thermoreversible hydrogel containing 200 mg of zarifremab, the AUC of zarifremab in urine is in the range of approximately 1.5 to 3 mg*hr / mL. 【0146】

[0146] In various embodiments, after a single intravesical administration of approximately 70 mL of thermoreversible hydrogel containing 300 mg of zarifremab, the AUC of zarifremab in urine is in the range of approximately 2.25 to 4.5 mg*hr / mL. 【0147】

[0147] In various embodiments, after a single intravesical administration of approximately 70 mL of thermoreversible hydrogel containing 400 mg of zarifremab, the AUC of zarifremab in urine is in the range of approximately 3 to 6 mg*hr / mL. 【0148】

[0148] In various embodiments, after a single intravesical administration of approximately 70 mL of thermoreversible hydrogel containing 500 mg of zarifremab, the AUC of zarifremab in urine is in the range of approximately 3.75 to 7.5 mg*hr / mL. 【0149】

[0149] In various embodiments, after a single intravesical administration of approximately 70 mL of thermoreversible hydrogel containing 600 mg of zarifremab, the AUC of zarifremab in urine is in the range of approximately 4.5 to 9 mg*hr / mL. 【0150】

[0150] In various embodiments, after a single intravesical administration of approximately 70 mL of thermoreversible hydrogel containing 700 mg of zarifremab, the AUC of zarifremab in urine is in the range of approximately 5.25 to 10.5 mg*hr / mL. 【0151】

[0151] In various embodiments, after a single intravesical administration of approximately 70 mL of the thermoreversible hydrogel of the present disclosure, the Cmax of zarifremab in urine is in the range of approximately 0.075–0.2 mg / mL, 0.15–0.4 mg / mL, 0.23–0.6 mg / mL, 0.3–0.8 mg / mL, 0.38–1.0 mg / mL, 0.45–1.2 mg / mL, and 0.5–1.4 mg / mL. 【0152】

[0152] In various embodiments, after a single intravesical administration of approximately 70 mL of thermoreversible hydrogel containing zarifremab, the t max It is found in urine over a period of approximately 2 to 12 hours. 【0153】

[0153] Anti-CTLA-4 antibodies have been shown to increase levels of immunomodulatory cytokines, including but not limited to IL-2, IL-6, and IL-8. Compositions of the Disclosure, administered topically (e.g., intravesically) and containing zarifremab, also achieve increases in levels of immunomodulatory cytokines (e.g., levels of one or more of IL-2, IL-4, IL-6, IL-8, IL-10, IFN-γ, and TNF-α) in bladder tissue, tumor tissue, and urine after administration of at least one dose of the composition according to the Disclosure. Levels of IL-2 that are more than twice that of untreated tissue correlate with a therapeutically effective dose of the composition according to the Disclosure. 【0154】

[0154] The composition of the Disclosure, which contains zarifremab and is administered topically (e.g., intravesically), shows that after approximately two or more doses (once a week) of the composition of the Disclosure, the CD4 levels are compared to the levels before administration of the composition of the Disclosure containing zarifremab. + ICOS hi T cell expression was increased. CD4 + ICOShi The increase in T cell expression ranges from approximately a 2x increase to more than a 10x increase (approximately 2x, 3x, 4x, 4x, 5x, 6x, 7x, 8x, 9x, 10x, or more than a 10x increase in CD4). + ICOS hi This includes an increase in T cell expression. 【0155】

[0155] The composition of the present disclosure, which is administered topically and contains zarifremab, is administered approximately two or more times (once a week) before CD4 + ICOS hi CD4 on the T side + FOXP3 + The ratio to T cells was also increased compared to the ratio before administration of the composition of this disclosure containing zarifremab. + ICOS hi / CD4 + FOX3P + The ratio is generally less than 1, and after administration of about two or more doses (once a week) of the composition of the Disclosure, the ratio increases by about two times or more (including about 2x times or more, about 3x times or more, about 4x times or more, or about 5x times or more, and including all ranges between any of these values). 【0156】

[0156] Various systemically administered anti-CTLA-4 antibodies have been shown to induce immune-related adverse events, such as skin lesions (rash, itching, vitiligo), colitis, hypophysitis, thyroiditis, sarcoidosis, uveitis, Guillain-Barré syndrome, immune-mediated cytopenia, and polymyalgia rheumatica / Horton's disease (Anne Bertrand et al., BMC Medicine (2015)). When anti-CTLA-4 antibodies are administered systemically, the overall incidence of immune-related adverse events is considerably high (over 70%), and the incidence of high-grade immune-related adverse events is 24%. Conversely, the composition of this disclosure containing zarifremab, administered intravesically, shows a significantly lower incidence of immune-related adverse events, including a lower incidence of high-grade immune-related adverse events. For example, when administered as described herein (e.g., intravesically, about once a week for about 6 to 12 weeks), the compositions of this disclosure exhibit an overall incidence of less than 70% of adverse events (including about 65%, about 60%, about 55%, about 50%, about 45%, about 40%, about 35%, about 30%, about 25%, about 20%, about 15%, or about 10%, and including all ranges between any of these values) and an overall incidence of less than 20% of high-grade adverse events (including about 20%, about 15%, about 10%, or about 5%, and including all ranges between any of these values). The incidence is assessed based on the number of systemic and specific immune-related adverse events (irAEs) (skin, gastrointestinal, endocrine, and hepatic disorders) and their grades (1-5; recorded according to versions 2, 3, or 4 of the National Cancer Institute's Common Terminology Criteria for Adverse Events). Grades ≥ 3 are considered high-grade. 【0157】

[0157] Systemically administered anti-CTLA-4 antibodies have been shown to increase levels of immunomodulatory cytokines such as IL-2, IL-4, IL-6, IL-8, IL-10, IFN-γ, and TNF-α. A composition of the Disclosure containing zarifremab administered topically / spotlessly (e.g., intravesically) may also result in an increase in levels of immunomodulatory cytokines (e.g., levels of one or more of IL-2, IL-4, IL-6, IL-8, IL-10, IFN-γ, and TNF-α) upon exposure to six doses of the composition of the Disclosure. The increase in cytokine levels correlates with the administration of a therapeutically effective dose of the composition of the Disclosure. 【0158】

[0158] The composition of the Disclosure, which contains zarifremab and is administered topically (e.g., intravesically), after approximately three or more doses (once a week) of the composition of the Disclosure, CD4 + ICOS hi T cell expression was increased compared to the level before administration of the composition of this disclosure containing zarifremab. CD4 + ICOS hi The increase in T cell expression is CD4 + ICOS hi In terms of T cell expression, the increase ranges from approximately twofold to more than tenfold (including increases of approximately twofold, threefold, fourfold, fourfold, fivefold, sixfold, sevenfold, eightfold, ninefold, tenfold, or more than tenfold). 【0159】

[0159] The composition of the present disclosure, which is administered topically and contains zarifremab, is administered approximately three or more times (once a week) before CD4 + ICOS hi CD4 on the T side + FOXP3 + The ratio to T cells was also increased compared to the ratio before administration of the composition of this disclosure containing zarifremab. + ICOS hi / CD4 + FOX3P +The ratio is generally less than 1, and after administration of approximately three or more doses (once a week) of the composition of the Disclosure, the ratio increases by approximately 2 times or more (including approximately 2x times or more, approximately 3x times or more, approximately 4x times or more, or approximately 5x times or more, and including all ranges between any of these values). 【0160】

[0160] To detect changes in cytokine levels in urine and blood, immunoassays or proximity extension assays can be performed. Immunoassays detect and measure cytokines using antibodies that antagonize each cytokine, along with colorimetric or electrochemiluminescence reactions. Non-limiting examples of this technique include, but are not limited to, ELISA or Mesoscale Discovery techniques. On the other hand, proximity extension assays (PEAs) utilize antibody pairs that antagonize each cytokine and contain a unique DNA sequence that enables hybridization only in the antibody pair. Subsequent proximity extension creates a DNA reporter sequence, which is amplified by a real-time polymerase chain reaction. A non-limiting example of a PEA is Olink. 【0161】

[0161] Changes in immune cell populations and immune cell markers are detected by immunohistochemistry of tumor tissue or by flow cytometry of immune cells in peripheral mononuclear cells (PBMCs) or urine. The concentration of zarifremab in human urine can be detected using immunoassays in combination with colorimetric or electrochemiluminescence. 【0162】

[0162] In a particular embodiment, at least about 80% of the zarifremab is released from the thermoreversible hydrogel within about 4 to about 24 hours after intravesical administration. 【0163】

[0163] In various embodiments, a thermoreversible hydrogel containing zarifremab is administered as monotherapy to treat bladder cancers, including, but not limited to, low-grade or high-grade nonmuscle-invasive bladder cancer, such as high-grade nonmuscle-invasive bladder cancer including papillary (Ta), lamina propria invasive (T1), and carcinoma in situ (CIS), muscle-invasive bladder cancer (MIBC), and metastatic bladder cancer. 【0164】

[0164] In some embodiments, a thermoreversible hydrogel containing zarifrelimab is prepared by mixing a solution of zarifrelimab with a thermoreversible hydrogel disclosed herein. In some embodiments, the zarifrelimab solution and the thermoreversible hydrogel are provided in the form of a kit containing separate vials of the zarifrelimab solution and the thermoreversible hydrogel. In certain embodiments, the zarifrelimab composition contains about 100 mg to about 750 mg of zarifrelimab in the thermoreversible hydrogel. In further specific embodiments, the volume of the composition administered to the patient is about 40 to 100 mL. In yet another further specific embodiment, the volume of the composition administered to the patient is about 70 mL. 【0165】

[0165] A thermoreversible hydrogel comprising 20% ​​to 32% (w / w) poloxamer and water, and approximately 100 to 900 mg of zarifremab is further provided herein. 【0166】

[0166] The pharmaceutical product described contains zarifremab at a concentration of 0.67 to 10 mg / mL, has a viscosity of 5000 cP or less at 5°C, and a viscosity of 1 × 10 at 19°C. 6 cP~9×10 7 It has a viscosity of cP. 【0167】

[0167] The pharmaceutical product described contains a thermoreversible hydrogel having a gelation temperature of approximately 10°C to approximately 18°C. 【0168】

[0168] The pharmaceutical product described contains a thermoreversible hydrogel having a gelation temperature of approximately 11°C to approximately 17°C. 【0169】

[0169] The pharmaceutical product described contains a thermoreversible hydrogel having a gelation temperature of approximately 12°C to 16°C. 【0170】

[0170] The pharmaceutical composition described contains a thermoreversible hydrogel having a gelation temperature of approximately 13°C to approximately 15°C. 【0171】

[0171] The pharmaceutical composition described contains poloxamer 407. 【0172】

[0172] The pharmaceutical composition described contains a thermoreversible hydrogel having at least a thickener, a stabilizer, an active oxygen scavenger, a surfactant, a buffer, and a gelation temperature regulator. 【0173】

[0173] The pharmaceutical composition described contains a thickener such as a cellulose derivative, a stabilizer such as sorbitol, an active oxygen scavenger such as methionine, a surfactant such as polysorbate 80, a buffer such as histidine buffer, and a gelation temperature regulator such as PEG-400. 【0174】

[0174] The pharmaceutical composition described contains a thickening agent such as HPMC. 【0175】

[0175] The pharmaceutical composition described comprises a thermoreversible hydrogel formulated for intravesical administration into the bladder, wherein at least about 80% of the zarifremab is released from the thermoreversible hydrogel within about 4 to about 12 hours. 【0176】

[0176] The pharmaceutical composition described, further comprising at least one additional therapeutic agent / activator. 【0177】

[0177] The pharmaceutical composition described contains at least one additional therapeutic agent such as mitomycin C, imiquimod, or gemcitabine. 【0178】

[0178] The listed pharmaceuticals contain mitomycin C at a concentration of approximately 0.2 mg / mL to approximately 2 mg / mL. 【0179】

[0179] The pharmaceutical composition described contains gemcitabine at a concentration of approximately 100 to 933 mg. 【0180】

[0180] The pharmaceutical composition described has a pH of approximately 6 to approximately 8. 【0181】

[0181] The pharmaceutical composition described yields a urinary concentration of zarifremab exceeding 20 ug / mL after a single intravesical administration of approximately 70 mL. 【0182】

[0182] The pharmaceutical composition described herein, when stored at approximately 2°C to 8°C for at least 24 hours, is substantially free of zarefrelimab aggregates after 24 hours. 【0183】

[0183] The pharmaceutical composition described herein is for use in the treatment of bladder cancer, and the thermoreversible hydrogel is administered intravesically into the bladder of the subject and remains in the bladder for at least 4 hours after administration, thereby resulting in a urinary concentration of at least 20-70 μg / mL of zarifremab in the urine of the subject. 【0184】

[0184] The pharmaceutical composition described increases cytokine levels in the subject for 2 to 24 hours after administration. 【0185】

[0185] This specification describes a kit for preparing a pharmaceutical composition, comprising (a) about 40 to 80 mL of a thermoreversible hydrogel containing poloxamer and water, and (b) about 10 to 20 mL of a solution containing 50 mg to 900 mg of zarifremab, wherein mixing (i) and (ii) forms about 50 to 100 mL of the pharmaceutical composition. 【0186】

[0186] A method for treating bladder cancer is further described herein, comprising the step of administering a pharmaceutical composition comprising a thermoreversible hydrogel containing 20% ​​to 32% (w / w) poloxamer and water and about 50 mg to about 900 mg of zarifrelimab to the bladder of a patient in need thereof, wherein the pharmaceutical composition substantially provides sustained release of zarifrelimab for about 4 to about 24 hours. 【0187】

[0187] The method described further comprises administering to a subject at least one additional therapeutic agent / activator, for example, imiquimod, mitomycin, or gemcitabine, or any mixture thereof. 【0188】

[0188] The method described comprises zarifremab and at least one additional therapeutic agent / activator, which are formulated and administered together in a thermoreversible hydrogel. 【0189】

[0189] The description includes the administration of zarifremab and additional therapeutic agents separately. 【0190】

[0190] The method described includes a pharmaceutical composition having an administered volume of approximately 50 mL to approximately 100 mL. 【0191】

[0191] The method described comprises a pharmaceutical composition administered to a patient at least six times at intervals of approximately one week. 【0192】

[0192] The method described comprises a pharmaceutical composition administered to a patient six times at one-week intervals. 【0193】

[0193] The method described includes the step of administering the pharmaceutical composition once a week for at least six times, and then administering the pharmaceutical composition every 12 weeks. 【0194】

[0194] The method described is for treating bladder cancer selected from high-grade or low-grade NMIBC or MIBC. 【0195】

[0195] A method for preparing a thermoreversible hydrogel is further described herein, comprising the step of mixing (a) a thermoreversible hydrogel containing poloxamer and water with (b) a solution containing zarifrerimab. 【0196】

[0196] The method described has a volume ratio of approximately 80 to approximately 20 (a) to (b). 【0197】

[0197] The method described contains a mixture in a volume of approximately 50 mL to approximately 100 mL. 【0198】 Pharmaceutical compositions in the form of thermoreversible hydrogels prepared by the method described in

[0198] are further described herein. 【0199】

[0199] A method for treating bladder cancer comprising the step of intravesically administering a pharmaceutical composition comprising a thermoreversible hydrogel containing poloxamer and water, and approximately 100 mg to approximately 900 mg of zarifremab, wherein the concentration of zarifremab in the pharmaceutical composition is 0.67 to 10 mg / mL, and the thermoreversible hydrogel has a viscosity of 1000 cP or less at 5°C and 1 × 10⁻⁶ at 19°C. 6 cP ~ 1.5 × 10 7 A method for treating bladder cancer is further described herein, wherein the hydrogel has a viscosity of cP and delivers an excretory concentration of at least 20-70 μg / mL of zarifremab into the urine of the subject, thereby treating the cancer. 【0200】

[0200] An additional method described herein is a method for inhibiting the regrowth of bladder cancer, comprising the step of intravesically administering a pharmaceutical composition comprising a thermoreversible hydrogel containing poloxamer and water and about 50 mg to about 900 mg of zarifremab, wherein the concentration of zarifremab in the pharmaceutical composition is 2.85 to 10 mg / mL, and the thermoreversible hydrogel has a viscosity of 1000 cP or less at 5°C and 1 × 10⁻⁶ at 19°C. 6 cP ~ 1.5 × 10 7This method inhibits bladder cancer regrowth by having a viscosity of cP, and the hydrogel delivers a urinary concentration of at least 20-70 μg / mL of zarifremab into the target urine, thereby inhibiting cancer regrowth. 【0201】

[0201] The method described comprises a pharmaceutical composition administered once a week for 3 to 9 weeks. 【0202】

[0202] A further method described herein is a method for reducing the size of a tumor before surgical resection of the tumor, comprising the step of intravesically administering to a subject a pharmaceutical composition comprising a thermoreversible hydrogel containing poloxamer and water and about 100 mg to about 750 mg of zarifrelimab, wherein the concentration of zarifrelimab in the pharmaceutical composition is 0.67 to 10 mg / mL, and the thermoreversible hydrogel has a viscosity of 400 cP or less at 5°C and 1 × 10⁻⁶ at 19°C. 6 cP ~ 1.5 × 10 7 This method reduces the size of a tumor before surgical resection, by having a viscosity of cP, and its hydrogel resulting in a urinary concentration of at least 20-70 μg / mL of zarifremab in the subject's urine, thereby shrinking the tumor before surgical resection. 【0203】

[0203] The method described includes a hydrogel administered once a week for 1 to 4 weeks. 【0204】

[0204] The method described further includes the step of intravesically administering an additional therapeutic agent selected from imiquimod, mitomycin C, or gemcitabine to the subject. 【0205】

[0205] The method described includes additional therapeutic agents administered before, simultaneously with, or after the administration of the pharmaceutical composition. 【0206】

[0206] 【0207】

[0207] The following examples are provided to illustrate certain features and / or embodiments. These examples should not be construed as limiting the disclosure to any specific features or embodiments described. [Examples] 【0208】

[0208] The following examples utilize the following components and follow the following parameters. 【0209】

[0209] Zarifremab is formulated as an intravesical solution called UGN-301 (supplied by Agenus Inc., Lexington, MA), which is a pharmaceutical composition containing zarifremab, an anti-CTLA4 antibody, and is formulated in a thermoreversible hydrogel for intravesical administration as described herein. Zarifremab is supplied in an amount between 100 and 700 mg in a volume of approximately 70 mL. The resulting supplied concentration range is 2.85 to 10 mg / mL. The concentration of zarifremab obtained in the urine produced is at least 20 to 80 μg / mL after at least 6 hours. UGN-301 is diluted with a thermoreversible hydrogel and diluent before administration to form the "UGN-301 mixture". 【0210】

[0210] Imiquimod (UGN-201) is a liquid formulation supplied at a dose of 200 mg (4 mg / mL) per 50 mL, administered intravesically, and retained in the bladder for approximately 60 minutes. 【0211】

[0211] Gemcitabine is supplied at a dose of 1000 mg (20 mg / mL) in 50 mL of liquid formulation, held in the bladder for at least 90 minutes, and then washed with saline solution (this is an additional regimen, and doses may be used, for example, up to 2000 mg, up to 100 mL in volume, and up to 2 hours in holding time). 【0212】

[0212] The administration regimen is administered once a week for 6 to 12 weeks in the induction schedule, and then in an optional maintenance schedule, as determined by an experienced clinician, it may be one of the following: 【0213】

[0213] Once every three months (at 6, 9, and 12 months after initial treatment), 【0214】

[0214] Administered for 3 weeks every 3 months (at 6, 9, and 12 months from the initial treatment). 【0215】

[0215] Administered every 3 months for 2 weeks (at 6, 9, and 12 months from the initial treatment). 【0216】

[0216] Once every 3 months (at 6, 9, and 12 months after initial treatment), then once every 6 months thereafter (at 18 and 24 months after initial treatment), 【0217】

[0217] Administer for 3 weeks every 3 months (at 6, 9, and 12 months from the initial treatment, and then for 3 weeks every 6 months thereafter (at 18 and 24 months from the initial treatment), or 【0218】

[0218] Administered for 2 weeks every 3 months (at 6, 9, and 12 months from the initial treatment, and then every 6 months for 2 weeks thereafter (at 18 and 24 months from the initial treatment). 【0219】

[0219] Composition of the AGEN1884 (zarefrelimab) drug product: The composition, including the amounts of all inactive components, is as follows. 【0220】

[0220] [Table 1] 【0221】

[0221] Concentration of AGEN1884 (zarefrelimab) excipient in the final mixture of UGN-301. The concentrations are in the range between their content in a 100 mg dose of UGN-301 and a 700 mg dose of UGN-301, as listed in the table below. 【0222】

[0222] [Table 2] 【0223】

[0223] AGEN1884 is extremely stable against aggregation. No increase in aggregates was detected in any sample at all ambient temperatures analyzed (even at high temperatures) up to 9 months. Thus, it can be seen here that AGEN1884 (zarefrelimab), when stored at approximately 2°C to 8°C for at least 24 hours, is substantially free of zarefrelimab aggregates after 24 hours. 【0224】

[0224] Hydrogel for UGN-301 composition: The content of the components in the hydrogel is listed below. 【0225】

[0225] [Table 3] 【0226】

[0226] Example 1 - Treatment with UGN-301 as monotherapy and combination therapy 【0227】

[0227] Systemic anti-CTLA-4 therapy has demonstrated clinical efficacy against solid tumors; however, systemic inhibition by CTLA-4 has also been associated with toxicity and adverse effects at doses delivered systemically, estimated to be in the range of 68–113 mg. These effects are presumed to be due to abnormal systemic immune activation. 【0228】

[0228] This specification describes the finding that intravesical administration of zarifremab in a thermoreversible hydrogel to the bladder wall successfully increased the drug concentration in the target organ without significant systemic exposure, thereby reducing systemic toxicity and adverse effects associated with CTLA-4 blockade. 【0229】

[0229] For a total of 27 patients with recurrent NMIBC, UGN-301 was administered intravesically as monotherapy or combination therapy, as described below. 【0230】

[0230] Monotherapy: A total of 20 patients were treated with UGN-301 alone as follows: 3 patients were treated with 100 mg of UGN-301 (i.e., 100 mg of zarifremab in the final formulation of UGN-301), 6 patients with 300 mg of UGN-301, 8 patients with 500 mg of UGN-301, and 3 patients with 700 mg of UGN-301. 【0231】

[0231] During the initiation period of monotherapy, patients received 70 mL of a UGN-301 mixture as a reverse-thermal hydrogel formulation once a week for 6 weeks, with a minimum of 4 days between treatments. Patients who were disease-free at 12 weeks and 6 months had the option of receiving maintenance therapy with 70 mL of an intravesical UGN-301 mixture as a reverse-thermal hydrogel formulation once a quarter, at 6 months, 9 months, and 12 months after the start of treatment. 【0232】

[0232] Combination treatment with UGN-201: Three patients were treated with 300 mg of UGN-301 and 200 mg of UGN-201, and one patient was treated with a combination of 500 mg of UGN-301 and 200 mg of UGN-201. 【0233】

[0233] During the UGN-301+UGN-201 initiation period, patients received intravesical instillation of 200 mg / 50 mL (4 mg / mL) of UGN-201 as a liquid formulation once a week for 6 weeks, held in the bladder for 1 hour and then emptied, followed by intravesical instillation of 70 mL of UGN-301 as a reverse-heat hydrogel formulation, with a minimum of 4 days between procedures. Patients who were disease-free at week 12 and month 6 had the option of receiving maintenance treatment quarterly at 6, 9, and 12 months after the start of treatment, consisting of 200 mg / 50 mL (4 mg / mL) of UGN-201 as a liquid formulation, held in the bladder for 1 hour and then emptied, followed by 70 mL of the intravesical UGN-301 mixture as a reverse-heat hydrogel formulation. 【0234】

[0234] Combination therapy with gemcitabine: Three patients were treated with a combination of 300 mg UGN-301 and 1000 mg gemcitabine. During the induction period, patients received intravesical instillation of 1000 mg / 50 mL (20 mg / mL) gemcitabine as a liquid formulation once a week for 6 weeks, held in the bladder for 90 minutes and then drained, followed by intravesical instillation of 70 mL of UGN-301 as a reverse-heat hydrogel formulation, with a minimum of 4 days between treatments. Patients who were disease-free at week 12 and month 6 had the option of receiving maintenance therapy quarterly at 6, 9, and 12 months after the start of treatment, consisting of 1000 mg / 50 mL (20 mg / mL) gemcitabine as a liquid formulation held in the bladder for 90 minutes and then drained, followed by 70 mL of the intravesical UGN-301 mixture as a reverse-heat hydrogel formulation. Between each drip infusion, the bladder was flushed with saline solution after the gemcitabine had been expelled, raising the pH inside the bladder before administering UGN-301 drip infusion. 【0235】

[0235] At all dose levels evaluated, the adverse events (TEAEs) that occurred during treatment were mild or moderate in severity, with the exception of two severe events, urinary tract infections, which were not considered to be related to the treatment or procedure of the study (Table 4). No patients experienced serious TEAEs or TEAEs that led to discontinuation of treatment. In addition, no dose-limiting toxicities or immune-related adverse events (irAEs) were observed. 【0236】

[0236] In two or more patients treated with UGN-301 as monotherapy, the following TEAEs were reported: hematuria, UTI, nausea, dysuria, and urinary retention, each occurring in two patients (Table 5). In two or more patients treated with UGN-301 and UGN-201, the reported TEAE was urinary incontinence, while in UGN-301 + gemcitabine, the reported TEAE was dysuria (Table 5). 【0237】

[0237] [Table 4] TIFF2026519989000006.tif128149 【0238】

[0238] N = number of patients in each treatment group; n = number of patients who experienced an event; TEAE = adverse event that occurred during treatment. 【0239】

[0239] a In the combined group, the TEAEs related to the treatments were caused by both treatments. 【0240】

[0240] Note: Percentages are based on the number of patients in each treatment group. 【0241】

[0241] [Table 5] 【0242】

[0242] N = number of patients in each treatment group; n = number of patients who experienced an event; TEAE = adverse event that occurred during treatment. 【0243】

[0243] a One patient had a severe or medically significant TEAE (Thirdly Epileptic Epileptic Examination) of urinary tract infection that the principal investigator considered to be related to the study procedure. 【0244】

[0244] Note: Percentages are based on the number of patients in each treatment group. 【0245】

[0245] Results 【0246】

[0246] In addition to not causing serious side effects, treatment with UGN-301 was effective in treating targeted bladder cancer at doses of 300 mg or higher. Half of the patients who received 300 mg of UGN-301 remained disease-free for more than 9 months after the start of treatment, and two-fifths of the patients who received 500 mg of UGN-301 remained disease-free for more than 3 months after the start of treatment. 【0247】

[0247] Furthermore, in patients who received a combination of 200 mg UGN-201 and 300 mg UGN-301 administered intrabladderally, all three patients who received this combination remained disease-free for three months after the start of treatment, indicating its effectiveness. 【0248】

[0248] In addition, it was found that treatment with a combination of 1000 mg UGN-201 and 300 mg UGN-301 administered intravesically was effective. As shown herein, two patients remained disease-free for three months after the start of treatment. 【0249】

[0249] These results are summarized in Table 6. 【0250】

[0250] [Table 6] 【0251】

[0251] Conclusion 【0252】

[0252] As can be seen from the above, monotherapy treatment of NMIBC using UGN-301 as monotherapy, treatment of NMIBC in combination with UGN-201, or treatment of NMIBC in combination with chemotherapeutic agents resulted in safe and effective treatment courses. Therefore, local administration of zarifremab into the bladder by intravesical administration using a thermoreversible hydrogel (UGN-301) allows for an increase in drug concentration in the target organ without significant systemic exposure, thereby reducing systemic toxicity and adverse effects associated with CTLA-4 blockade, which demonstrates its advantage over systemic administration for the treatment of NMIBC. 【0253】

[0253] Example 2 - Pharmacokinetics of treatment with UGN-301 【0254】

[0254] Serum samples were collected from weeks 1 to 6 prior to administration, and at 1, 2, 6, and 12 hours (±15 minutes) and 24 hours (±1 hour) after UGN-301 administration in weeks 1 and 6, and the concentration of circulating zarifrimab was measured. Samples taken from 16 patients (14 patients received 100-500 mg of UGN-301 alone, and 2 patients received a combination of 300 mg of UGN-301 and 200 mg of UGN-201) showed no systemic exposure to zarifrimab after single and repeated doses of UGN-301 administered intravesically once weekly for up to 6 weeks. The sustained concentrations of zarifrimab detected in urine over time were below the level of quantification, indicating no exposure. This data suggests that intravesical administration of zarifrimab likely reduced systemic adverse events. 【0255】

[0255] Data on anti-zalifrimab antibodies in seven patients who received UGN-301 at doses of 100-500 mg showed that no antibodies antagonizing zarifrimab were produced, and therefore, it did not affect systemic exposure to zarifrimab. 【0256】

[0256] Urinary concentrations were analyzed in patients who received 100, 300, or 500 mg of UGN-301 once weekly (Table 7). Maximum urinary concentrations occurred 2 hours after administration of UGN-301 at all dose levels, and were 197+ / -77, 627+ / -174, and 1106+ / -314 μg / mL for 100, 300, and 500 mg of UGN-301, respectively. Zarifremab was detectable in the urine for at least 12 hours at all evaluated dose levels, and in some patients, it was detectable up to 22-24 hours after administration of 300 and 500 mg of UGN-301. These data demonstrate that the maximum urinary concentration and AUC increase proportionally with doses of approximately 100-500 mg. Based on the available data, the maximum inhibition of CTLA-4 occurs in the range of 4.7–14.7 hours when the time exceeds the threshold concentration (70 μg / mL) (Figure 1 and Table 7). 【0257】

[0257] 【Table 7】 【0258】 【0258】This example demonstrates that the local administration of UGN-301 has successfully achieved sustained release of zalifrelimab that can be detected in the urine of patients up to 24 hours after administration. 【0259】 【0259】 Changes in cytokine expression resulting from treatment with Example 3 - UGN-301 【0260】 【0260】Using the Olink proximity extension assay (PEA) technology, changes in 92 proteins, including various cytokines in urine, after intravesical administration of 100 mg or 300 mg of UGN-301 were evaluated. Overall, the maximum changes occurred within the first 12 hours, and most proteins had returned to or were returning to baseline by 24 hours. Treatment with UGN-301 resulted in changes in abundance for 89 / 92 proteins at any time and at any dose. Evaluation of the proteins that most consistently increased among patients and doses revealed that the cytokines HO-1, CASP-8, CCL20, and ARG1 increased by more than 2 log2-fold between 2 and 24 hours after treatment with 100 or 300 mg of UGN-301. The cytokines GAL-9, TRAIL, ADGRG1, IFN-γ, and CXCL9 increased by more than 2 log2-fold between 2 and 24 hours after treatment with 300 mg of UGN-301. The cytokines CCL3, CCL4, GZMA, and IL18 increased by more than 2 log2-fold between 2 and 24 hours after treatment with 100 mg of UGN-301. Taken together, these data indicate that local administration of UGN-301 leads to a rapid increase in immunomodulatory cytokines. 【0261】 【0261】 Example 4 - Monotherapy - Prevention of Recurrence 【0262】 【0262】As described in Example 1, subjects who have undergone cystectomy to remove a tumor are given an intravesical regimen of UGN-301 at an initial dose of 100 mg, and optionally escalated to 700 mg of UGN-301 to reach a concentration of zarefrelimab in urine of 20 - 80 μg / mL and maintain that for at least 6 hours. UGN-301 is administered weekly for at least 6 weeks. This is found to prevent the regrowth of tumors after surgical resection. 【0263】 【0263】Subjects will optionally be treated during a maintenance period, during which the patient will receive up to 3 additional doses of the treatment assigned to that patient once every 3 months (at 6, 9, and 12 months after the start of treatment). 【0264】 【0264】 Example 5 - Monotherapy - Neoadjuvant Treatment 【0265】 【0265】As described in Example 1, subjects who are prior to undergoing cystectomy are given an intravesical regimen of 100 mg of the antibody in UGN-301, and optionally escalated to 700 mg of the antibody in UGN-301 to maintain a concentration of zarefrelimab in urine of 20 - 80 μg / mL for at least 6 hours. UGN-301 is administered weekly for 1 - 4 weeks prior to surgery, and this treatment is found to reduce tumor size prior to surgery. 【0266】 【0266】Subjects will optionally be treated during a maintenance period, during which the patient will receive up to 3 additional doses of the treatment assigned to that patient once every 3 months (at 6, 9, and 12 months after the start of treatment). 【0267】 【0267】 Example 6 - Combination Therapy (UGN-301 + UGN-201) - Prevention of Recurrence 【0268】

[0268] This example is similar to Example 1, except that the subjects are administered a combination of zarifremab and imiquimod, rather than UGN-301 monotherapy. Subjects requiring it are given a combination regimen administered intravesically weekly for at least 6 weeks, in which 200 mg / 50 mL (4 mg / mL) of UGN-201 as a liquid formulation is held in the bladder for 1 hour, then flushed / washed, followed by 100-700 mg of UGN-301 (zarifremab) formulated in a thermoreversible hydrogel, maintaining a urinary zarifremab concentration of 20-80 μg / mL for at least 6 hours. This is shown to prevent tumor regrowth after surgical resection of the tumor. 【0269】

[0269] Example 7 - Combination therapy (UGN-301 + UGN-201) - Neoadjuvant treatment 【0270】

[0270] This example is similar to Example 1, except that the subjects are administered a combination of zarifrimab and imiquimod, rather than UGN-301 monotherapy. Subjects requiring this treatment are administered the combination regimen intravesically for 1 to 4 weeks weekly, in which 200 mg / 50 mL (4 mg / mL) of UGN-201 as a liquid formulation is held in the bladder for 1 hour and then excreted, followed by 100 to 700 mg of UGN-301 (zarifrimab) formulated in a thermoreversible hydrogel, maintaining a urinary zarifrimab concentration of 20 to 80 μg / mL for at least 6 hours. This treatment is shown to reduce tumor size prior to surgical resection of the tumor(s) or bladder. 【0271】

[0271] Example 8 - Combination therapy (UGN-301 + gemcitabine) - Complete response 【0272】

[0272] This example is similar to Example 1, except that the subjects are administered a combination of zarifremab and gemcitabine, rather than UGN-301 monotherapy. Subjects requiring this regimen are administered intravesically weekly for at least 6 weeks, in which 1000 mg / 50 mL (20 mg / mL) of gemcitabine is held in the bladder for 90 minutes and then drained, followed by saline lavage, then an initial dose of 100 mg of UGN-301, followed by escalation of UGN-301 formulated in a thermoreversible hydrogel, possibly up to 700 mg, to maintain a urinary zarifremab concentration of 20-80 μg / mL for at least 6 hours. This procedure is found to lead to the disappearance of all detectable tumor lesions using techniques established in the art. 【0273】

[0273] Example 9 - Combination therapy (UGN-301 + gemcitabine) - Prevention of relapse 【0274】

[0274] This example is similar to Example 1, except that the subjects are administered a combination of zarifremab and gemcitabine, rather than UGN-301 monotherapy. Subjects requiring it are given the combination regimen intravesically every week for at least 6 weeks, in which 1000 mg / 50 mL (20 mg / mL) of gemcitabine is held in the bladder for 90 minutes and then excreted, followed by saline irrigation, then an initial dose of 100 mg of UGN-301, followed by gradual increases of UGN-301 formulated in a thermoreversible hydrogel, up to 700 mg if necessary, to maintain a urinary zarifremab concentration of 20-80 μg / mL for at least 6 hours. This treatment is shown to prevent tumor regrowth after surgical resection of the tumor. 【0275】

[0275] Example 10 - Combination therapy (UGN-301 + gemcitabine) - Neoadjuvant treatment 【0276】

[0276] This example is similar to Example 1, except that the subjects are administered a combination of zarifrimab and gemcitabine, rather than UGN-301 monotherapy. For subjects requiring it, the combination regimen is administered intravesically for 1 to 4 weeks weekly, in which 1000 mg / 50 mL (20 mg / mL) of gemcitabine is held in the bladder for 90 minutes and then drained, followed by saline irrigation, then an initial dose of 100 mg of UGN-301, followed by gradual increases of UGN-301 formulated in a thermoreversible hydrogel, up to 700 mg if necessary, to maintain a urinary zarifrimab concentration of 20-80 μg / mL for at least 6 hours. This treatment is found to reduce tumor size prior to surgical resection of the tumor(s) or bladder. 【0277】

[0277] Example 11 - Combination therapy (UGN-301 + Mitomycin C) complete response 【0278】

[0278] This example is similar to Example 1, except that the subjects are administered a combination of zarifremab and mitomycin C, rather than UGN-301 monotherapy. Subjects requiring this treatment are administered the combination regimen intravesically weekly for 6 weeks, in which 75 mg of mitomycin C and 200-700 mg of UGN-301 (zarifremab) are formulated together in a thermoreversible hydrogel to maintain a urinary zarifremab concentration of 20-80 μg / mL for at least 6 hours. This treatment is found to lead to the disappearance of all detectable tumor lesions using techniques established in the art. 【0279】

[0279] Example 12 - Combination therapy (UGN-301 + Mitomycin C) - Prevention of relapse 【0280】 【0280】This example is similar to Example 1, except that instead of monotherapy with UGN-301, a combination of zalifrelimab and mitomycin C is administered to the subject. The required subjects are given a combination regimen intravesically for 6 weeks, weekly, in which 75 mg of mitomycin C and 200 - 700 mg of UGN-301 (zalifrelimab) are formulated together in a thermoreversible hydrogel to maintain the concentration of zalifrelimab in the urine at 20 - 80 μg / mL for at least 6 hours. This treatment is found to prevent tumor regrowth after surgical resection of the tumor. 【0281】 【0281】 Example 13 - Combination Therapy (UGN-301 + Mitomycin C) - Neoadjuvant Treatment 【0282】 【0282】This example is similar to Example 1, except that instead of monotherapy with UGN-301, a combination of zalifrelimab and mitomycin C is administered to the subject. The required subjects are given a combination regimen intravesically for 1 - 4 weeks, weekly, in which 75 mg of mitomycin C and 200 - 700 mg of UGN-301 (zalifrelimab) are formulated together in a thermoreversible hydrogel to maintain the concentration of zalifrelimab in the urine at 20 - 80 μg / mL for at least 6 hours. This treatment is found to reduce tumor size prior to surgical resection of the tumor(s) or the bladder.

Claims

[Claim 1] i) Viscosity of 5000 cP or less at 5°C, and 1 × 10⁻¹⁶ at 17°C. ６ cP ~ 9 × 10 ７ A thermoreversible hydrogel having a viscosity of cP, ii) Zarefrelimab liquid solution and A pharmaceutical composition containing a mixture of the following. [Claim 2] The pharmaceutical composition according to claim 1, wherein the thermoreversible hydrogel contains poloxamer. [Claim 3] The pharmaceutical composition according to claim 2, wherein the thermoreversible hydrogel contains 25% to 32% poloxamer. [Claim 4] The pharmaceutical composition according to claim 2, wherein the thermoreversible hydrogel contains a thickening agent. [Claim 5] The pharmaceutical composition according to claim 3, wherein the thermoreversible hydrogel contains 20% to 32% (w / w) poloxamer, and the thickening agent is HPMC. [Claim 6] The pharmaceutical composition according to claim 2, wherein the thermoreversible hydrogel comprises 25% to 32% poloxamer, 0.02% to 0.5% HPMC, 0.75% to 1% PEG, and water. [Claim 7] The pharmaceutical composition according to claim 1, wherein the zarefrelimab liquid solution is formulated in a solution containing histidine, sorbitol, methionine, and polysorbate 80, and is adjusted to a pH of 6 to 7. [Claim 8] The pharmaceutical composition according to claim 7, wherein the zarefrelimab liquid solution is formulated in a solution containing 10-30 mM histidine, 200-300 mM sorbitol, 10-20 mM methionine, and 0.01-0.03% polysorbate 80, and is adjusted to a pH of 6-7. [Claim 9] The pharmaceutical composition according to claim 1, wherein the zarifremab is present in a dose of 100 mg to 750 mg. [Claim 10] The pharmaceutical composition according to claim 1, wherein the zarifremab has a concentration of 0.67 to 10 mg / mL. [Claim 11] The pharmaceutical composition according to claim 1, wherein the composition is formulated for intravesical administration into the bladder, and at least about 80% of the detectable zarifremab is released from the thermoreversible hydrogel within about 2 to about 12 hours. [Claim 12] A pharmaceutical composition according to any one of claims 1 to 11, further comprising at least one additional therapeutic agent. [Claim 13] The pharmaceutical composition according to claim 12, wherein the additional therapeutic agent is mitomycin C, imiquimod, or gemcitabine. [Claim 14] The pharmaceutical composition according to claim 13, wherein the concentration of imiquimod is about 1 mg / mL to about 10 mg / mL. [Claim 15] The pharmaceutical composition according to claim 13, wherein the dose of gemcitabine is approximately 100 mg to 3000 mg. [Claim 16] The pharmaceutical composition according to claim 15, wherein the dose of gemcitabine is up to about 2000 mg. [Claim 17] The pharmaceutical composition according to claim 13, wherein the concentration of mitomycin C is about 0.1 mg / mL to about 4 mg / mL. [Claim 18] The pharmaceutical composition according to any one of claims 1 to 17, wherein, after a single intravesical administration of the zarifremab, the detectable urinary concentration of zarifremab exceeds 20 μg / mL. [Claim 19] The pharmaceutical composition according to any one of claims 1 to 18, which, when stored at approximately 2°C to 8°C for at least 24 hours, substantially does not contain aggregates of zarefrelimab after 24 hours. [Claim 20] The pharmaceutical composition according to any one of claims 1 to 19, wherein the thermoreversible hydrogel further comprises at least a stabilizer, an active oxygen scavenger, a surfactant, a buffer, and a gelation temperature regulator. [Claim 21] The pharmaceutical composition according to any one of claims 1 to 20, wherein the thermoreversible hydrogel has a gelation temperature of about 10°C to about 20°C, particularly 13°C to about 16°C. [Claim 22] The pharmaceutical composition according to any one of claims 1 to 21, wherein the poloxamer is poloxamer 407. [Claim 23] The pharmaceutical composition according to any one of claims 1 to 22, wherein the pH of the pharmaceutical composition is about 6 to about 8. [Claim 24] A pharmaceutical composition according to any one of claims 1 to 23, for use in the treatment of bladder cancer, wherein the thermoreversible hydrogel is administered intravesically into the bladder of a subject, thereby resulting in a urinary concentration of at least 20 to 70 μg / mL of zarifremab in the urine of the subject at least two hours after administration. [Claim 25] The pharmaceutical composition for use according to claim 24, wherein the urinary tract cancer is bladder cancer. [Claim 26] The pharmaceutical composition for use according to claim 25, wherein the bladder cancer is high-grade or low-grade NMIBC, MIBC, or carcinoma in situ (CIS). [Claim 27] A kit for preparing a pharmaceutical composition, i. A thermoreversible hydrogel containing 20% ​​to 32% poloxamer 407, ii. A solution containing 100 mg to 750 mg of zarifremab, Includes, A kit in which mixing (i) and (ii) forms approximately 20 to 100 mL of the aforementioned pharmaceutical composition. [Claim 28] A kit for preparing a pharmaceutical composition, i. 60-73 mL of thermoreversible hydrogel containing 20-32% poloxamer 407, ii. A 2-15 mL solution containing 100 mg to 750 mg of zarifremab, Includes, A kit in which mixing (i) and (ii) forms approximately 75 mL of the aforementioned pharmaceutical composition. [Claim 29] A kit for preparing a pharmaceutical composition, i. Approximately 40-80 mL of thermoreversible hydrogel, ii. A mixture of approximately 1 to 10 mL of a stabilizer, an active oxygen scavenger, a surfactant, a buffer, and a gelling temperature regulator, iii. A solution containing 50 mg to 750 mg of zarifremab in approximately 10 to 20 mL, Includes, A kit in which mixing (i), (ii), and (iii) forms approximately 50 to 100 mL of the aforementioned pharmaceutical composition. [Claim 30] A ready-to-use pharmaceutical composition for administration to a patient, comprising zarifrerimab, poloxamer, stabilizer, buffer, reactive oxygen species scavenger, and gelling temperature regulator. [Claim 31] The pharmaceutical composition according to claim 1, wherein the thermoreversible hydrogel comprises 25% to 32% poloxamer, 0.02% to 0.5% HPMC, and water.

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