Bicyclic peptide ligands specific for EphA2 and uses thereof

Abstract

The invention relates to a bicyclotoxin conjugate BT5528 or a pharmaceutically acceptable salt thereof, a pharmaceutical composition of the bicyclotoxin conjugate BT5528 and an application of the bicyclotoxin conjugate BT5528.

Description

Technical Field

[0001] This invention relates to bicyclic toxin conjugates or pharmaceutically acceptable salts thereof, or pharmaceutical compositions thereof. The invention also provides the use of bicyclic toxin conjugates or pharmaceutically acceptable salts thereof, or pharmaceutical compositions thereof, for the prevention or treatment of diseases, conditions, or disorders characterized by EphA2 overexpression in diseased tissues. Background of the Invention

[0003] Cyclic peptides can bind to protein targets with high affinity and target specificity, making them an attractive class of molecules for therapeutic development. In fact, several cyclic peptides (such as the antimicrobial peptide vancomycin, the immunosuppressant cyclosporine, or the anticancer drug octreotide) have been successfully used clinically (Driggers et al. (2008), Nat Rev Drug Discov). 7 (7), 608-24). The favorable binding properties stem from the relatively large interaction surface formed between the peptide and the target, and the reduced conformational flexibility of the cyclic structure. Typically, macrocyclic compounds bind to surfaces of hundreds of square angstroms, such as the cyclic peptide CXCR4 antagonist CVX15 (400 Å). 2 ; Wu et al. (2007), Science 330, 1066-71 (a protein with integrin αVb3 (355 Å) 2 Cyclic peptides with Arg-Gly-Asp motifs that bind to urokinase-type plasminogen activators (Xiong et al. (2002), Science 296 (5565), 151-5)) or cyclic peptide inhibitors upain-1 (603 Å) that bind to urokinase-type plasminogen activators. 2 ; Zhao et al. (2007), J Struct Biol 160 (1), 1-10).

[0004] Due to their cyclic configuration, macrocyclic peptides are less flexible than linear peptides, resulting in less entropy loss during target binding and thus higher binding affinity. The reduced flexibility also leads to target-specific conformation locking compared to linear peptides, enhancing binding specificity. This effect has been demonstrated by potent and selective inhibitors of matrix metalloproteinase 8 (MMP-8), which lose their selectivity relative to other MMPs when their rings are opened (Cherney et al. (1998), J MedChem 41 (11), 1749-51). In polycyclic peptides with more than one peptide ring, such as vancomycin, nisin, and actinomycin, the advantageous binding properties gained through macrocyclization are even more pronounced.

[0005] Different research groups have previously linked peptides with cysteine ​​residues to synthetic molecular structures (Kemp and McNamara (1985), J. Org. Chem; Timmerman et al. (2005), ChemBioChem). Meloen and colleagues used tris(bromomethyl)benzene and related molecules to rapidly and quantitatively cyclize multiple peptide rings onto synthetic scaffolds for structural simulation of protein surfaces (Timmerman et al. (2005), ChemBioChem). Methods for generating candidate drug compounds, wherein the compounds are generated by linking cysteine-containing peptides to molecular scaffolds such as TATA (1,1',1''-(1,3,5-triazinane-1,3,5-triyl)tripropyl-2-en-1-one, Heinis et al., Angew Chem, Int Ed. 2014; 53:1602–1606).

[0006] Combinatorial methods based on phage display have been developed to generate and screen large libraries of bicyclic peptides targeting targets of interest (Heinis et al. (2009), Nat Chem Biol). 5 (7), 502-7 and WO 2009 / 098450). In short, a combinatorial library of linear peptides containing three cysteine ​​residues and two six-random-amino acid regions (Cys-(Xaa)6-Cys-(Xaa)6-Cys) was displayed on phages and cyclized by covalently linking the cysteine ​​side chains to a small molecule scaffold. Invention Overview

[0008] According to one aspect, the present invention provides a method of treating a human patient, comprising administering BT5528 or a pharmaceutical composition containing BT5528.

[0009] On the other hand, the present invention provides a method for treating solid tumors in human patients, comprising administering BT5528 or a pharmaceutical composition containing BT5528. The present invention also provides a method for using BT5528 or a pharmaceutical composition containing BT5528 to treat solid tumors in human patients.

[0010] On the other hand, the present invention provides a method for maintaining or reducing the volume of a solid tumor in a human patient, comprising administering BT5528 or a pharmaceutical composition containing BT5528 to the patient.

[0011] In another aspect, the present invention provides a method for treating a patient with a solid tumor using BT5528 or a pharmaceutical composition comprising BT5528, wherein the method comprises administering BT5528 or a pharmaceutical composition thereof to the patient.

[0012] In another aspect, the present invention provides a method for maintaining or reducing the volume of a solid tumor in a patient using BT5528 or a pharmaceutical composition containing BT5528, wherein the method comprises administering BT5528 or a pharmaceutical composition containing BT5528 to the patient.

[0013] In another aspect, the present invention provides BT5528 or a pharmaceutically acceptable salt thereof for use in a method of manufacturing a medicament for treating a solid tumor in a patient, wherein the method comprises administering BT5528 to the patient.

[0014] In another aspect, the present invention provides the use of BT5528 or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for maintaining or reducing the volume of a solid tumor in a human patient, wherein the method comprises administering BT5528 to the patient.

[0015] Detailed description of certain implementation schemes

[0016] 1. Compounds and definitions: The term "BT5528" as used herein refers to a bicyclic toxin conjugate having the structure shown below, wherein the molecular scaffold is 1,1',1''-(1,3,5-triazinane-1,3,5-triyl)tripropyl-2-en-1-one (TATA), and the peptide ligand contains the following amino acid sequence: (β-Ala)-Sar 10 -A(HArg)DC i (HyP)LVNPLC ii LHP(D-Asp)W(HArg)C iii (SEQ ID NO:1) Sar is creatine, HArg is arginine, and HyP is hydroxyproline.

[0017] As used herein, the term "pharmaceutically acceptable salt" means that, to the extent of reliable medical judgment, it is suitable for use in contact with tissues of humans and lower animals without excessive toxicity, irritation, or allergic reactions, and is commensurate with a reasonable benefit / risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, pharmaceutically acceptable salts are described in detail by SM Berge et al. in J. Pharmaceutical Sciences, 1977, 66, 1–19 (incorporated herein by reference). Pharmaceutically acceptable salts of the compounds of the present invention include those derived from suitable inorganic and organic acids and bases. Examples of pharmaceutically acceptable non-toxic acid addition salts are amino salts formed with inorganic acids (such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, and perchloric acid) or with organic acids (such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid, or malonic acid), or amino salts formed by other methods used in the art, such as ion exchange. Other pharmaceutically acceptable salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, hydrogen sulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, disglucuronate, dodecyl sulfate, ethanesulfonate, formate, fumarate, glucohepanoate, glycerophosphate, glucuronate, hemisulfate, heptaate, hexanoate, hydroiodide, 2-hydroxyethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, dihydroxynaphthalate, pectate, persulfate, 3-phenylpropionate, phosphate, neopentanoate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate, etc.

[0018]

[0019] Salts derived from suitable bases include alkali metals, alkaline earth metals, ammonium, and nitrogen. + (C 1–4 Alkyl)4 salts. Representative alkali metal or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, etc. Where appropriate, other pharmaceutically acceptable salts include non-toxic ammonium, quaternary ammonium, and amine cations that form counterions using halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, lower alkyl sulfonates, and aryl sulfonates. It should be understood that the form of the salt is within the scope of this invention, and references to peptide ligands include the salt form of said ligand.

[0020] The salts of the present invention can be produced from a parent compound containing a basic or acidic moiety through conventional chemical methods (such as...). Pharmaceutical Salts:Properties, Selection, and Use(See P. Heinrich Stahl (ed.), Camille G. Wermuth (ed.), ISBN: 3-90639-026-8, Hardcover, 388 pages, August 2002, for the method described). Typically, such salts can be prepared by reacting the free acid or base form of these compounds with a suitable base or acid in water or an organic solvent or a mixture of both.

[0021] As used herein, the term "about" should have the meaning of being within 10% of a given value or range. In some implementations, the term "about" refers to being within 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, or 1% of a given value.

[0022] Unless otherwise stated, the structures described herein also imply all isomers (e.g., enantiomers, diastereomers, and geometric (or conformations)) of that structure; for example, R and S configurations for each asymmetric center, Z and E double bond isomers, and Z and E conformational isomers. Therefore, single stereochemical isomers of the compounds of the present invention, as well as enantiomers, diastereomers, and mixtures of geometric (or conformations), are within the scope of the present invention. Unless otherwise stated, all tautomers of the compounds of the present invention are within the scope of the present invention. Furthermore, unless otherwise stated, the structures described herein are also intended to include compounds that differ only in the presence of one or more isotopically enriched atoms. For example, compounds having the structures of the present invention and including substitutions of deuterium or tritium for hydrogen, or... 13 C or 14 Compounds enriched with carbon that substitute for carbon are within the scope of this invention. According to this invention, such compounds can be used, for example, as analytical tools, as probes in bioassays, or as therapeutic agents.

[0023] 2. Treatment methods

[0024] On one hand, the present invention provides a method or use for treating solid tumors in human patients. In some embodiments, the solid tumor is an advanced malignant tumor, including administering BT5528 to the patient or administering a pharmaceutical composition containing BT5528 to the patient. In some embodiments, the solid tumor or advanced tumor is associated with EphA2 expression.

[0025] In some embodiments, the method of the present invention includes administering BT5528. In some embodiments, the method of the present invention includes administering a pharmaceutically acceptable salt of BT5528. In some embodiments, the method of the present invention includes administering a pharmaceutical composition comprising BT5528 or a pharmaceutically acceptable salt thereof.

[0026] In some embodiments, the present invention provides a method for maintaining or reducing the volume of a solid tumor in a patient. In some embodiments, the present invention provides a method for maintaining or reducing the volume of a solid tumor in a patient, wherein the method includes administering BT5528 or a pharmaceutically acceptable salt thereof. In some embodiments, the present invention provides a method for maintaining or reducing the volume of a solid tumor in a patient, wherein the method includes administering a pharmaceutical composition comprising BT5528.

[0027] In some implementations, maintaining the volume of a solid tumor means an increase in tumor volume of no more than 75%, 50%, 25%, 10%, or 5%. In some implementations, maintaining the volume of a solid tumor means keeping the tumor volume unchanged.

[0028] In some implementations, reducing the volume of a solid tumor means reducing the volume of the tumor by 100%, at least 95%, at least 90%, at least 85%, at least 80%, at least 75%, or at least 70%, at least 65%, at least 60%, at least 55%, at least 50%, at least 45%, at least 40%, at least 35%, at least 30%, at least 25%, at least 20%, at least 15%, at least 10%, or at least 5%.

[0029] Optionally, the change in tumor volume is relative to the change in tumor volume before the start of treatment.

[0030] In some embodiments, references to tumor volume may refer to the sum of the volumes of two or more tumors in a patient (e.g., two, three, four, five, six, seven, eight, nine, or ten or more tumors). In some embodiments, this can be determined by measuring the sum of the axes (e.g., the short axis or the longest diameter) of two or more solid tumors in the patient.

[0031] In some embodiments, references to tumor volume may refer to the average volume of two or more tumors in a patient (e.g., two, three, four, five, six, seven, eight, nine, or ten or more tumors). In some embodiments, this can be determined by measuring the sum of the axes (e.g., the minor axis or the longest diameter) of two or more solid tumors in a patient (e.g., two, three, four, five, six, seven, eight, nine, or ten or more tumors) and dividing that sum by the number of tumors measured.

[0032] Therefore, in some implementations, maintaining or reducing the volume of solid tumors in a patient, as described herein, may refer to maintaining or reducing the total or average volume of two or more solid tumors in the patient (e.g., two, three, four, five, six, seven, eight, nine, or ten or more tumors).

[0033] Therefore, in some embodiments, maintaining the volume of a solid tumor means that the total or average volume of two or more tumors (e.g., two, three, four, five, six, seven, eight, nine, or ten or more tumors) increases by no more than 75%, no more than 50%, no more than 25%, no more than 10%, or no more than 5%. In some embodiments, maintaining the volume of a solid tumor means that the total or average volume of the tumor remains unchanged.

[0034] In some implementations, reducing the volume of a solid tumor means reducing the total or average volume of two or more tumors (e.g., two, three, four, five, six, seven, eight, nine, or ten or more tumors) by 100%, at least 95%, at least 90%, at least 85%, at least 80%, at least 75%, or at least 70%, at least 65%, at least 60%, at least 55%, at least 50%, at least 45%, at least 40%, at least 35%, at least 30%, at least 25%, at least 20%, at least 15%, at least 10%, or at least 5%.

[0035] Optionally, the change in the total or average volume of the tumor is relative to the change in the total or average volume of the tumor before the start of treatment.

[0036] The volume of the tumor (or multiple tumors) may be maintained or reduced for a period of time until disease progression is recorded. In some embodiments, disease progression is defined as a tumor volume that has increased by more than 75% relative to the tumor volume before treatment began. In some embodiments, disease progression is defined as a measurement of the tumor's axial length (or the sum or average of two or more tumors) that has increased by more than 20%. Optionally, the tumor volume may be maintained or reduced for at least 8 weeks, at least 9 weeks, at least 10 weeks, at least 11 weeks, at least 12 weeks, at least 13 weeks, at least 14 weeks, at least 15 weeks, at least 16 weeks, at least 17 weeks, at least 18 weeks, at least 19 weeks, at least 20 weeks, at least 6 months, at least 9 months, at least 12 months, at least 15 months, at least 18 months, at least 21 months, or at least 24 months.

[0037] In some embodiments, the volume of a solid tumor over time can be measured by determining the axial length of the tumor. In some embodiments, the volume of a solid tumor over time can be measured by determining the length of its maximum diameter. In some embodiments, the volume of a solid tumor over time can be measured by determining its short axis. As a representative embodiment, a 20% increase in the axial length or diameter of the solid tumor corresponds to an increase in the solid tumor volume of approximately 75% (i.e., 1.2). 3 ).

[0038] The axial length of a tumor can be determined by means of, for example, CT scan sections, MRI scan sections, caliper measurements, or X-rays.

[0039] In other implementations, the volume of a solid tumor over time can be assessed by volumetric evaluation, for example, by using three-dimensional scanning (such as CT or MRI scans).

[0040] In some implementations, treatment of the cancer or solid tumor may result in a complete remission of the cancer or solid tumor to the treatment, i.e., the disappearance of all target lesions and all non-target lesions and a return to normal tumor marker levels. Optionally, treatment of the cancer or solid tumor may result in a partial remission of the cancer or solid tumor to the treatment, i.e., a reduction of at least 30% in the sum of the axial lengths (e.g., the longest diameter (LD)) of the target lesions, with reference to the baseline sum of the axial lengths (e.g., the longest diameter). Optionally, treatment of the cancer or solid tumor may result in stable disease, i.e., a reduction of no more than 30% in the sum of the axial lengths (e.g., LD) but an increase of less than 20% in the sum of the axial lengths (e.g., LD), with no new lesions.

[0041] 3. Combined treatment

[0042] In some embodiments, the present invention provides the combined use of BT5528 and an anti-PD-1 antibody or an anti-PD-L1 antibody for the treatment of solid tumors. In some embodiments, the treatment method of the present invention includes administering BT5528 or a pharmaceutical composition containing BT5528 to a human patient, followed by further administration of an anti-PD-1 antibody or an anti-PD-L1 antibody.

[0043] In some implementations, the anti-PD-1 antibody is selected from the following list: pembrolizumab, nivolumab, cimiprimab, dotalimab, rivalimab, vopalimumab, spartazumab, camrelizumab, sintilimab, tagorimab, toripalimab, INCMGA00012, AMP-224, AMP-514, and acriximab.

[0044] In some implementations, the anti-PD-L1 antibody is selected from the following list: atezolizumab, avelumumab, durvalumab, KN035, and cosibelimab.

[0045] In some embodiments, the method of the present invention includes administering BT5528 or a pharmaceutical composition containing BT5528 to a human patient, and further administering nivolumab.

[0046] Nivolumab can be administered as described on the label, which can be found at https: / / www.opdivohcp.com / dosing / dosing-schedules, the contents of which are incorporated herein by reference in their entirety. In some embodiments, 240 mg of nivolumab is administered every 2 weeks. In some embodiments, 480 mg of nivolumab is administered every 4 weeks. In some embodiments, nivolumab is administered via intravenous infusion over 30 minutes.

[0047] In some embodiments of combined use, the patient's intolerance to immune checkpoint inhibitors was not previously known. In some embodiments of combined use, the patient's allergy to checkpoint inhibitor therapy was not known. In some embodiments of combined use, the patient had not previously undergone organ transplantation. In some embodiments of combined use, the patient had not previously been diagnosed with clinically relevant immunodeficiency. In some embodiments of combined use, the patient did not have an active systemic infection requiring treatment. In some embodiments of combined use, the patient was taking no more than 10 mg daily of a prednisone equivalent or other strong immunosuppressant. In some embodiments of combined use, the patient had no history of autoimmune diseases other than alopecia or vitiligo. In some embodiments of combined use, the patient had no history of interstitial lung disease.

[0048] In some embodiments of combined use, BT5528 or a pharmaceutical composition containing BT5528 is administered separately from nivolumab. In some embodiments, BT5528 or a pharmaceutical composition containing BT5528 is administered sequentially with nivolumab. In some embodiments, BT5528 or a pharmaceutical composition containing BT5528 is administered first, followed by nivolumab. In some embodiments, nivolumab is administered first, followed by BT5528 or a pharmaceutical composition containing BT5528.

[0049] In some embodiments, BT5528 and nivolumab are administered via separate infusions. In some embodiments, BT5528 and nivolumab are administered via sequential infusions. In some embodiments, BT5528 is administered via a first infusion, followed by nivolumab. In some embodiments, nivolumab is administered via a first infusion, followed by BT5528.

[0050] 4. Overview of certain embodiments of the present invention

[0051] In some embodiments, the method of the present invention includes administering a pharmaceutical composition comprising BT5528 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient or carrier. In some embodiments, the pharmaceutical composition comprises about 21.2 mg of BT5528 or a pharmaceutically acceptable salt thereof.

[0052] In some embodiments, the pharmaceutical composition is a solid pharmaceutical composition. In some embodiments, the solid pharmaceutical composition of the present invention is a powder. In some embodiments, the pharmaceutical composition of the present invention is a lyophilized powder. In some embodiments, the solid pharmaceutical composition of the present invention is a granule.

[0053] In some embodiments, the pharmaceutical composition is a liquid pharmaceutical composition. In some embodiments, the liquid pharmaceutical composition of the present invention is a pharmaceutical formulation in an acceptable medium or solvent. In some embodiments, the acceptable medium or solvent is selected from sterile water, Ringer's solution, USP, and isotonic sodium chloride solution. In some embodiments, the acceptable medium or solvent is sterile water. In some embodiments, the acceptable medium or solvent is a sterile injection medium. In some embodiments, the liquid pharmaceutical composition contains about 2-4 mg / mL BT5528 or a pharmaceutically acceptable salt thereof.

[0054] In some embodiments, pharmaceutically acceptable excipients or carriers include buffers. In some embodiments, the buffer is selected from phosphates, glycine, sorbic acid, potassium sorbate, mixtures of glycerides of saturated vegetable fatty acids, water, salts, or electrolytes, such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinylpyrrolidone, cellulose-based materials, polyethylene glycol, sodium carboxymethyl cellulose, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, polyethylene glycol, and lanolin. In some embodiments, the buffer is histidine hydrochloride. In some embodiments, the buffer is sodium hydroxide. In some embodiments, the buffer is hydrochloric acid.

[0055] In some embodiments, the amount of buffer can adjust the pH of the pharmaceutical composition of the present invention to about 6-8. In some embodiments, the buffer is histidine hydrochloride in an amount of about 1-3 mg / mg BT5528 or a pharmaceutically acceptable amount thereof. In some embodiments, the amount of histidine hydrochloride is about 1.31 or 2.62 mg / mg BT5528 or a pharmaceutically acceptable salt thereof. In some embodiments, the liquid pharmaceutical composition of the present invention comprises histidine hydrochloride at a concentration of about 5.24 mg / mL.

[0056] In some embodiments, the pH value of the liquid pharmaceutical composition of the present invention is about 6-8. In some embodiments, the pH value of the liquid pharmaceutical composition of the present invention is about 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, or 8.0. In some embodiments, the pH value of the liquid pharmaceutical composition of the present invention is about 6.5 or 7.0.

[0057] In some embodiments, the pharmaceutically acceptable excipient or carrier includes an inert pharmaceutically acceptable excipient or carrier. In some embodiments, the inert pharmaceutically acceptable excipient or carrier is sodium citrate or dicalcium phosphate. In some embodiments, the inert pharmaceutically acceptable excipient or carrier is a filler or expander. In some embodiments, the filler or expander is starch, lactose, sucrose, glucose, mannitol, or silicic acid. In some embodiments, the inert pharmaceutically acceptable excipient or carrier is a binder. In some embodiments, the binder is carboxymethyl cellulose, alginate / ester, gelatin, polyvinylpyrrolidone, sucrose, or acacia. In some embodiments, the inert pharmaceutically acceptable excipient or carrier is selected from sucrose, trehalose, glucose, or combinations thereof. In some embodiments, the inert pharmaceutically acceptable excipient or carrier is sucrose.

[0058] In some embodiments, the amount of inert, pharmaceutically acceptable excipient or carrier (e.g., sucrose) is about 10-35 mg / mg of BT5528 or a pharmaceutically acceptable salt thereof. In some embodiments, the amount of inert, pharmaceutically acceptable excipient or carrier (e.g., sucrose) is about 15 or 30 mg / mg of BT5528 or a pharmaceutically acceptable salt thereof. In some embodiments, the liquid pharmaceutical composition comprises an inert, pharmaceutically acceptable excipient or carrier (e.g., sucrose) at a concentration of about 60 mg / mL.

[0059] In some embodiments, the pharmaceutically acceptable excipient or carrier comprises a surfactant. In some embodiments, the surfactant is a polysorbate (e.g., polysorbate-20, polysorbate-40, polysorbate-60, polysorbate-65, polysorbate-80, polysorbate-85, or combinations thereof). In some embodiments, the surfactant is selected from poloxamer (e.g., poloxamer 188); Triton™; sodium lauryl sulfate (SDS); sodium lauryl sulfate; sodium octyl glycoside; lauryl-sulfobetaine, myristyl-sulfobetaine, linoleyl-sulfobetaine, stearyl-sulfobetaine, lauryl-sarcosine, myristyl-sarcosine, linoleyl-sarcosine, stearyl-sarcosine, linoleyl-betaine, myristyl-betaine, cetyl The surfactants include methyl betaine, lauramidopropyl betaine, cocamidopropyl betaine, linoleamide-propyl betaine, myristamidopropyl betaine, palmitamidopropyl betaine, isostearamidopropyl betaine (e.g., lauramidopropyl), myristamidopropyl dimethylamine, palmitamidopropyl dimethylamine, or isostearamidopropyl dimethylamine; sodium methylcocoyl taurate or disodium methyloleyl taurate; and the Monaquat™ series (Mona Industries, Inc., Paterson, NJ), polyethylene glycol, polypropylene glycol, and copolymers of ethylene and propylene glycol (e.g., pluronics, PF68). In some embodiments, the surfactant is polysorbate 20.

[0060] In some embodiments, the amount of surfactant (e.g., polysorbate 20) is about 0.01-0.15 mg / mg BT5528, or a pharmaceutically acceptable salt thereof. In some embodiments, the amount of surfactant (e.g., polysorbate 20) is about 0.025, 0.05, or 0.1 mg / mg BT5528, or a pharmaceutically acceptable salt thereof. In some embodiments, the liquid pharmaceutical composition comprises a surfactant (e.g., polysorbate 20) at a concentration of about 0.1 or 0.2 mg / mL.

[0061] In some embodiments, the pharmaceutically acceptable excipient or carrier comprises an isotropic modifier. In some embodiments, the isotropic modifier is sodium chloride, glucose, calcium chloride, or a combination thereof. In some embodiments, the isotropic modifier is glucose. In some embodiments, the isotropic modifier is sodium chloride. In some embodiments, the isotropic modifier is a combination of sodium chloride and glucose.

[0062] In some embodiments, the present invention provides a pharmaceutical composition comprising BT5528 or a pharmaceutically acceptable salt thereof, histidine hydrochloride, sucrose, and polysorbate 20. In some embodiments, the pharmaceutical composition of the present invention comprises: BT5528 or a pharmaceutically acceptable salt thereof; Approximately 1.31–2.62 mg histidine hydrochloride / mg BT5528 or a pharmaceutically acceptable salt thereof; Approximately 15-30 mg sucrose / mg BT5528 or a pharmaceutically acceptable salt thereof; and Approximately 0.05-0.1 mg of polysorbate 20 / mg BT5528 or a pharmaceutically acceptable salt thereof.

[0063] In some embodiments, the present invention provides a solid pharmaceutical composition, which is a lyophilized powder comprising: Approximately 21.2 mg of BT5528 or a pharmaceutically acceptable salt thereof; Approximately 55.5 mg histidine hydrochloride / mg BT5528 or a pharmaceutically acceptable salt thereof; Approximately 636 mg sucrose / mg BT5528 or a pharmaceutically acceptable salt thereof; and Approximately 1.06–2.12 mg of polysorbate 20 / mg BT5528 or a pharmaceutically acceptable salt thereof.

[0064] In some embodiments, the present invention provides a solid pharmaceutical composition, which is a lyophilized powder comprising: Approximately 21.2 mg of BT5528 or a pharmaceutically acceptable salt thereof; Approximately 27.8 mg histidine hydrochloride / mg BT5528 or a pharmaceutically acceptable salt thereof; Approximately 318 mg sucrose / mg BT5528 or a pharmaceutically acceptable salt thereof; and Approximately 1.06 mg of polysorbate 20 / mg BT5528 or a pharmaceutically acceptable salt thereof.

[0065] In some embodiments, the present invention provides a liquid pharmaceutical composition comprising: Approximately 2-4 mg / mL BT5528 or a pharmaceutically acceptable salt thereof; Approximately 5.25 mg / mL of histidine hydrochloride; Approximately 60 mg / mL of sucrose; and Polysorbate 20 at approximately 0.1-0.2 mg / mL.

[0066] In some embodiments, the present invention provides a liquid pharmaceutical composition prepared by dissolving the solid pharmaceutical composition of the present invention in water. In some embodiments, the present invention provides a liquid pharmaceutical composition prepared by dissolving the solid pharmaceutical composition of the present invention in an injectable medium (e.g., saline or 5% glucose). In some embodiments, the present invention provides a liquid pharmaceutical composition prepared by reconstituted the solid pharmaceutical composition of the present invention in water, followed by dilution with 5% glucose. In some embodiments, the liquid pharmaceutical composition is diluted into a 5% glucose IV bag for intravenous administration. In some embodiments, the liquid pharmaceutical composition in the 5% glucose IV bag is stored at room temperature (about 20-25°C) for up to about 4 hours prior to IV administration. In some embodiments, the liquid pharmaceutical composition in the 5% glucose IV bag is stored under refrigeration (about 2-8°C) for up to about 20 hours prior to IV administration. In some embodiments, the liquid pharmaceutical composition in the 5% glucose IV bag is stored under refrigeration (about 2-8°C) for up to about 20 hours prior to IV administration, followed by storage at room temperature (about 20-25°C) for up to about 4 hours.

[0067] In some embodiments, the present invention provides a solid pharmaceutical composition comprising BT5528 or a pharmaceutically acceptable salt thereof, histidine hydrochloride, sucrose, polysorbate 20, and glucose. In some embodiments, the present invention provides a liquid pharmaceutical composition comprising BT5528 or a pharmaceutically acceptable salt thereof, histidine hydrochloride, sucrose, polysorbate 20, glucose, and water. In some embodiments, the amounts, concentrations, and proportions of the components of the pharmaceutical composition are as described above.

[0068] In some embodiments, the method of the present invention includes intravenous administration of the pharmaceutical composition described herein to a patient. In some embodiments, the pharmaceutical composition of the present invention is administered by intravenous injection. In some embodiments, the pharmaceutical composition of the present invention is administered by intravenous infusion. In some embodiments, the intravenous infusion of the pharmaceutical composition of the present invention lasts for about 5-30 minutes. In some embodiments, the intravenous infusion of the pharmaceutical composition of the present invention lasts for about 30-90 minutes. In some embodiments, the intravenous infusion of the pharmaceutical composition of the present invention lasts for about 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, or 90 minutes. In some embodiments, the intravenous infusion of the pharmaceutical composition of the present invention lasts for about 60 minutes. In some embodiments, the intravenous infusion of the pharmaceutical composition of the present invention lasts for about 2, 2.5, 3, 3.5, or 4 hours.

[0069] 5. Dosing regimen

[0070] In some embodiments, BT5528 or a pharmaceutical composition containing BT5528 is administered to the patient every 1, 2, 3, 4, 5, 6, or 7 days. In some embodiments, the pharmaceutical composition of the present invention is administered to the patient weekly. In some embodiments, the pharmaceutical composition of the present invention is administered to the patient every two weeks.

[0071] In some embodiments, the treatment period is 4 weeks, or 28 days. In some embodiments, BT5528 is administered weekly during the 28-day treatment period, for example, on days 1, 8, 15, and 22. In some embodiments, BT5528 is administered every two weeks during the 28-day period (e.g., on days 1 and 15). In some embodiments, BT5528 is administered as a liquid formulation or in liquid unit dosage form.

[0072] In some implementations, the treatment cycle is 3 weeks, or a 21-day treatment cycle. In some implementations, BT5528 is administered during weeks 1 and 2 of the 21-day treatment cycle, but not during week 3. In some implementations, BT5528 is administered on days 1 and 8 of the 21-day treatment cycle. In some implementations, BT5528 is administered weekly during the 21-day treatment cycle. In some implementations, BT5528 is administered on days 1, 8, and 15 of the 21-day treatment cycle.

[0073] In some implementations, at approximately 1-27 mg / m 2 BT5528 is administered at a dose (measured by the amount of API: BT5528). In some embodiments, it is administered at approximately 2-20 mg / m². 2 BT5528 is administered at a dose of approximately 2-15 mg / m². In some embodiments, this is done in a dose of approximately 2-15 mg / m². 2 BT5528 is administered at a dose of approximately 2-10 mg / m². In some embodiments, this is done in a dose of approximately 2-10 mg / m². 2 BT5528 is administered at a dose of approximately 4-8 mg / m². In some embodiments, this is done in a dose of approximately 4-8 mg / m². 2 The dosage, optionally at about 5-7 mg / m², is... 2 The dosage, optionally at about 6.5 mg / m², is... 2 BT5528 is administered at a dose of approximately 3-7 mg / m². In some embodiments, this is done in a dose of approximately 3-7 mg / m². 2 The dosage, optionally at about 4-6 mg / m² 2 The dosage, optionally at about 5 mg / m², is [optional]. 2 BT5528 is administered at doses of approximately 2.2, 4.4, 7.3, 11, 14.6, or 19.4 mg / m³. In some embodiments, this is done at doses of approximately 2.2, 4.4, 7.3, 11, 14.6, or 19.4 mg / m³. 2BT5528 is administered at doses of approximately 1.5-3.5, 3.5-5.5, 6.5-8.5, 10-12, 13.5-15.5, or 18.5-20.5 mg / m². 2 BT5528 is administered at a dose of approximately 1-10 or 10-20 mg / m². In some embodiments, this is done at a dose of approximately 1-10 or 10-20 mg / m². 2 BT5528 is administered at doses of approximately 21, 22, 23, 24, 25, 26, or 27 mg / m². 2 BT5528 was administered at the prescribed dosage.

[0074] In some embodiments, the method of the present invention includes administering approximately 4-8 mg / m² every two weeks. 2 The dosage is optional, approximately 5-7 mg / m². 2 The dosage is optional, approximately 6.5 mg / m². 2 BT5528 or a pharmaceutical composition containing BT5528 is administered to the patient at the prescribed dose.

[0075] In some embodiments, the method of the present invention includes administering approximately 3-7 mg / m² weekly. 2 The dosage, optionally about 4-6 mg / m², optionally about 5 mg / m² 2 BT5528 or a pharmaceutical composition containing BT5528 is administered to the patient at the prescribed dose.

[0076] In some embodiments, the present invention provides a method for treating solid tumors in human patients, comprising administering approximately 4-8 mg / m² every two weeks. 2 The dosage is optional, approximately 5-7 mg / m². 2 The dosage is optionally approximately 6.5 mg / m². 2 BT5528 was administered to the patient at the prescribed dose.

[0077] In some embodiments, the present invention provides a method for treating solid tumors in human patients, comprising administering approximately 3-7 mg / m² weekly. 2 The dosage is optional, approximately 4-6 mg / m². 2 The dosage is optionally about 5 mg / m². 2 BT5528 was administered to the patient at the prescribed dose.

[0078] In some embodiments, the method of the present invention includes administering approximately 6.5 mg / m² every two weeks. 2 BT5528 or a pharmaceutical composition containing BT5528 is administered to the patient at the prescribed dose.

[0079] In some embodiments, the method of the present invention includes administering approximately 5 mg / m³ weekly. 2BT5528 or a pharmaceutical composition containing BT5528 is administered to the patient at the prescribed dose.

[0080] In some embodiments, the present invention provides a method for treating solid tumors in human patients, comprising administering approximately 6.5 mg / m² every two weeks. 2 BT5528 was administered to the patient at the prescribed dose.

[0081] In some embodiments, the present invention provides a method for treating solid tumors in human patients, comprising administering approximately 5 mg / m² weekly. 2 BT5528 was administered to the patient at the prescribed dose.

[0082] In some embodiments, the present invention provides a method for treating urothelial carcinoma in human patients, comprising administering approximately 4-8 mg / m² every two weeks. 2 The dosage is optional, approximately 5-7 mg / m². 2 The dosage is optionally approximately 6.5 mg / m². 2 The patient is given BT5528 or a pharmaceutical composition containing BT5528 at a specified dose. In some embodiments, the urothelial carcinoma is EphA2 positive, i.e., associated with EphA2 expression.

[0083] In some embodiments, the present invention provides a method for treating urothelial carcinoma in human patients, comprising administering approximately 3-7 mg / m² weekly. 2 The dosage is optional, approximately 4-6 mg / m². 2 The dosage is optional, approximately 5 mg / m 2 The dosage of BT5528 or a pharmaceutical composition containing BT5528 is administered to the patient. In some embodiments, the urothelial carcinoma is EphA2 positive, i.e., associated with EphA2 expression.

[0084] In some embodiments, the present invention provides a method for treating urothelial carcinoma in human patients, comprising administering approximately 6.5 mg / m² every two weeks. 2 The patient is given BT5528 or a pharmaceutical composition containing BT5528 at a specified dose. In some embodiments, the urothelial carcinoma is EphA2 positive, i.e., associated with EphA2 expression.

[0085] In some embodiments, the present invention provides a method for treating urothelial carcinoma in human patients, comprising administering approximately 5 mg / m² weekly. 2 The patient is given BT5528 or a pharmaceutical composition containing BT5528 at a specified dose. In some embodiments, the urothelial carcinoma is EphA2 positive, i.e., associated with EphA2 expression.

[0086] In some embodiments, the present invention provides a method for treating ovarian cancer in human patients, comprising administering approximately 4-8 mg / m² every two weeks. 2 The dosage is optional, approximately 5-7 mg / m². 2 The dosage is optionally approximately 6.5 mg / m². 2 The ovarian cancer is administered to the patient at a dose of BT5528 or a pharmaceutical composition containing BT5528. In some embodiments, the ovarian cancer is EphA2 positive, i.e., associated with EphA2 expression.

[0087] In some embodiments, the present invention provides a method for treating ovarian cancer in human patients, comprising administering approximately 3-7 mg / m² weekly. 2 The dosage is optional, approximately 4-6 mg / m². 2 The dosage is optional, approximately 5 mg / m 2 The ovarian cancer is administered to the patient at a dose of BT5528 or a pharmaceutical composition containing BT5528. In some embodiments, the ovarian cancer is EphA2 positive, i.e., associated with EphA2 expression.

[0088] In some embodiments, the present invention provides a method for treating ovarian cancer in human patients, comprising administering approximately 6.5 mg / m² every two weeks. 2 The patient is given BT5528 or a pharmaceutical composition containing BT5528 at a specified dose. In some embodiments, the ovarian cancer is EphA2 positive, i.e., associated with EphA2 expression.

[0089] In some embodiments, the present invention provides a method for treating ovarian cancer in human patients, comprising administering approximately 5 mg / m² weekly. 2 The patient is given BT5528 or a pharmaceutical composition containing BT5528 at a specified dose. In some embodiments, the ovarian cancer is EphA2 positive, i.e., associated with EphA2 expression.

[0090] In some embodiments, the present invention provides a method for treating non-small cell lung cancer (NSCLC) in human patients, comprising administering approximately 4-8 mg / m² every two weeks. 2 The dosage is optional, approximately 5-7 mg / m². 2 The dosage is optionally approximately 6.5 mg / m². 2 The dosage of BT5528 or a pharmaceutical composition containing BT5528 is administered to the patient. In some embodiments, the NSCLC is EphA2 positive, i.e., associated with EphA2 expression.

[0091] In some embodiments, the present invention provides a method for treating NSCLC in human patients, comprising administering approximately 3-7 mg / m² weekly. 2The dosage is optional, approximately 4-6 mg / m². 2 The dosage is optional, approximately 5 mg / m 2 The NSCLC is administered to the patient at a dose containing BT5528 or a pharmaceutical composition containing BT5528. In some embodiments, the NSCLC is EphA2 positive, i.e., associated with EphA2 expression.

[0092] In some embodiments, the present invention provides a method for treating NSCLC in human patients, comprising administering approximately 6.5 mg / m² every two weeks. 2 The NSCLC is administered to the patient at a dose of BT5528 or a pharmaceutical composition containing BT5528. In some embodiments, the NSCLC is EphA2 positive, i.e., associated with EphA2 expression.

[0093] In some embodiments, the present invention provides a method for treating NSCLC in human patients, comprising administering approximately 5 mg / m² weekly. 2 The NSCLC is administered to the patient at a dose of BT5528 or a pharmaceutical composition containing BT5528. In some embodiments, the NSCLC is EphA2 positive, i.e., associated with EphA2 expression.

[0094] In some embodiments, the present invention provides a method for treating head and neck cancer in human patients, comprising administering approximately 4-8 mg / m² every two weeks. 2 The dosage is optional, approximately 5-7 mg / m². 2 The dosage is optionally approximately 6.5 mg / m². 2 The dosage of BT5528 or a pharmaceutical composition containing BT5528 is administered to the patient. In some embodiments, the head and neck cancer is EphA2 positive, i.e., associated with EphA2 expression.

[0095] In some embodiments, the present invention provides a method for treating head and neck cancer in human patients, comprising administering approximately 3-7 mg / m² weekly. 2 The dosage is optional, approximately 4-6 mg / m². 2 The dosage, optionally at about 5 mg / m², is [optional]. 2 The dosage of BT5528 or a pharmaceutical composition containing BT5528 is administered to the patient. In some embodiments, the head and neck cancer is EphA2 positive, i.e., associated with EphA2 expression.

[0096] In some embodiments, the present invention provides a method for treating head and neck cancer in human patients, comprising administering approximately 6.5 mg / m² every two weeks. 2 The patient is given BT5528 or a pharmaceutical composition containing BT5528 at a specified dose. In some embodiments, the head and neck cancer is EphA2 positive, i.e., associated with EphA2 expression.

[0097] In some embodiments, the present invention provides a method for treating head and neck cancer in human patients, comprising administering approximately 5 mg / m² weekly. 2 The patient is given BT5528 or a pharmaceutical composition containing BT5528 at a specified dose. In some embodiments, the head and neck cancer is EphA2 positive, i.e., associated with EphA2 expression.

[0098] In some embodiments, the present invention provides a method for treating gastric cancer or upper gastrointestinal cancer in human patients, comprising administering approximately 4-8 mg / m² every two weeks. 2 The dosage is optional, approximately 5-7 mg / m². 2 The dosage is optionally approximately 6.5 mg / m². 2 Administer BT5528 to the patient at the prescribed dosage. 或 A pharmaceutical composition comprising BT5528. In some embodiments, the gastric cancer or upper gastrointestinal cancer is EphA2 positive, i.e., associated with EphA2 expression.

[0099] In some embodiments, the present invention provides a method for treating gastric cancer or upper gastrointestinal cancer in human patients, comprising administering approximately 3-7 mg / m² weekly. 2 The dosage is optional, approximately 4-6 mg / m². 2 The dosage is optional, approximately 5 mg / m 2 The patient is given BT5528 or a pharmaceutical composition containing BT5528 at a specified dose. In some embodiments, the gastric cancer or upper gastrointestinal cancer is EphA2 positive, i.e., associated with EphA2 expression.

[0100] In some embodiments, the present invention provides a method for treating gastric cancer or upper gastrointestinal cancer in human patients, comprising administering approximately 6.5 mg / m² every two weeks. 2 The patient is given BT5528 or a pharmaceutical composition containing BT5528 at a specified dose. In some embodiments, the gastric cancer or upper gastrointestinal cancer is EphA2 positive, i.e., associated with EphA2 expression.

[0101] In some embodiments, the present invention provides a method for treating gastric cancer or upper gastrointestinal cancer in human patients, comprising administering approximately 5 mg / m² weekly. 2 The patient is given BT5528 or a pharmaceutical composition containing BT5528 at a specified dose. In some embodiments, the gastric cancer or upper gastrointestinal cancer is EphA2 positive, i.e., associated with EphA2 expression.

[0102] In some embodiments, the present invention provides a method for treating triple-negative breast cancer (TNBC) in human patients, comprising administering approximately 4-8 mg / m² every two weeks.2 The dosage is optional, approximately 5-7 mg / m². 2 The dosage is optionally approximately 6.5 mg / m². 2 The TNBC is administered to the patient at a dose of BT5528 or a pharmaceutical composition containing BT5528. In some embodiments, the TNBC is EphA2 positive, i.e., associated with EphA2 expression.

[0103] In some embodiments, the present invention provides a method for treating TNBC in human patients, comprising administering approximately 3-7 mg / m² weekly. 2 The dosage is optional, approximately 4-6 mg / m². 2 The dosage is optional, approximately 5 mg / m 2 The TNBC is administered to the patient at a dose of BT5528 or a pharmaceutical composition containing BT5528. In some embodiments, the TNBC is EphA2 positive, i.e., associated with EphA2 expression.

[0104] In some embodiments, the present invention provides a method for treating TNBC in human patients, comprising administering approximately 6.5 mg / m² every two weeks. 2 The TNBC is administered to the patient at a dose of BT5528 or a pharmaceutical composition containing BT5528. In some embodiments, the TNBC is EphA2 positive, i.e., associated with EphA2 expression.

[0105] In some embodiments, the present invention provides a method for treating TNBC in human patients, comprising administering approximately 5 mg / m² weekly. 2 The TNBC is administered to the patient at a dose of BT5528 or a pharmaceutical composition containing BT5528. In some embodiments, the TNBC is EphA2 positive, i.e., associated with EphA2 expression.

[0106] In some embodiments, the method of treating solid tumors of the present invention provides a way to maintain or reduce the volume of a solid tumor in a patient. In some embodiments, the cancer is TNBC. In some embodiments, the cancer is NSCLC. In some embodiments, the cancer is gastric cancer or upper gastrointestinal cancer. In some embodiments, the cancer is head and neck cancer. In some embodiments, the cancer is ovarian cancer.

[0107] In some embodiments, the present invention provides a method for maintaining or reducing the volume of a solid tumor in a human patient, including administering approximately 5 mg / m² weekly. 2 The invention provides a method for maintaining or reducing the volume of a solid tumor in a human patient, comprising administering BT5528 at a dose of approximately 6.5 mg / m² every two weeks. 2The patient is given BT5528 at the specified dose. In some embodiments, the cancer is TNBC. In some embodiments, the cancer is NSCLC. In some embodiments, the cancer is gastric cancer or upper gastrointestinal cancer. In some embodiments, the cancer is head and neck cancer. In some embodiments, the cancer is ovarian cancer.

[0108] 6. Patients with solid tumors

[0109] In some embodiments, the solid tumor or advanced malignant tumor is selected from lung cancer (e.g., NSCLC), ovarian cancer, fallopian tube cancer, breast cancer (e.g., TNBC), head and neck cancer, gastric / upper gastrointestinal (GI) cancer, pancreatic cancer, urothelial carcinoma (e.g., bladder cancer, urethral cancer, urethral bladder cancer, and urothelial carcinoma), bone cancer, rectal cancer, gastric cancer, and squamous cell carcinoma of unknown origin. In some embodiments, the solid tumor or advanced malignant tumor associated with EphA2 expression is an adenocarcinoma subtype of NSCLC (adeno-NSCLC).

[0110] In one embodiment, the solid tumor or advanced malignant tumor is lung cancer. In one embodiment, the lung cancer is NSCLC.

[0111] In one embodiment, the solid tumor or advanced malignant tumor is ovarian cancer. In another embodiment, the solid tumor or advanced malignant tumor is fallopian tube cancer.

[0112] In one embodiment, the solid tumor or advanced malignant tumor is breast cancer. In one embodiment, the breast cancer is triple-negative breast cancer.

[0113] In one implementation, the solid tumor or advanced malignant tumor is head and neck cancer.

[0114] In one embodiment, the solid tumor or advanced malignant tumor is gastric cancer. In another embodiment, the solid tumor or advanced malignant tumor is upper gastrointestinal (GI) cancer.

[0115] In one implementation, the solid tumor or malignant tumor is pancreatic cancer.

[0116] In one embodiment, the solid tumor or malignant tumor is urothelial carcinoma. In one embodiment, the urothelial carcinoma is bladder cancer. In one embodiment, the urothelial carcinoma is urethral cancer. In one embodiment, the urothelial carcinoma is urethral-bladder cancer. In one embodiment, the urothelial carcinoma is urothelial carcinoma.

[0117] In one implementation, the solid tumor or advanced malignant tumor is gastric cancer.

[0118] In one implementation, the solid tumor or advanced malignant tumor is rectal cancer.

[0119] In one implementation, the solid tumor or advanced malignant tumor is squamous cell carcinoma of unknown origin.

[0120] In some implementations, the solid tumor or advanced malignant tumor is associated with EphA2 expression.

[0121] In some implementations, BT5528 is administered to patients who are at least 18 years of age. In other words, in some implementations, the patient is an adult.

[0122] In some implementations, the patient is a patient with an Eastern Cooperative Oncology Group (ECOG) performance status score of 0 or 1. ECOG performance status scores of 0 and 1 are described in Example 1.

[0123] In some implementations, the patient is a patient with measurable disease according to the Evaluation Criteria for Treatment of Solid Tumors (RECIST) version 1.1.

[0124] In some embodiments, the patient is a patient with acceptable organ function. In some embodiments, a patient with acceptable organ function has laboratory data selected from the following: Renal function: Creatinine clearance ≥50 mL / min, calculated according to the Cockcroft-Gault formula or measured by 24-hour urine collection; Total bilirubin ≤1.5 × ULN (upper limit of normal); Serum albumin ≥2.5 g / dL; In the presence of liver metastases, aspartate aminotransferase (AST) ≤2.5 × ULN or ≤5 × ULN; In the presence of liver metastases, alanine aminotransferase levels are ≤2.5 × ULN or ≤5 × ULN; and International Normalized Ratio (INR) < 1.3 or ≤ institutional ULN (contraindicated anticoagulant).

[0125] In some embodiments, the patient is a patient with acceptable hematological function. In some embodiments, a patient with acceptable hematological function has laboratory data selected from the following: Hemoglobin ≥9 g / dL; Absolute neutrophil count (ANC) ≥ 1500 cells / mm 3 ;and Platelet count ≥75,000 cells / mm 3 .

[0126] In some embodiments, the patient does not receive red blood cell or platelet transfusions or use growth factors within 4 weeks of the first administration of the pharmaceutical composition of the present invention.

[0127] In some implementations, the patient is a fertile female patient (WOCBP) with a negative pregnancy test (negative serum test at screening and negative urine or serum test within 3 days prior to the first administration of BT5528).

[0128] In some implementations, the patient has a histologically confirmed metastatic or recurrent malignant solid tumor, has exhausted all appropriate treatment options according to local guidelines, must have experienced failure in at least one previous first-line treatment, and has evidence of radiographic progression in the most recent first-line treatment.

[0129] In some implementations, the patient has previously received immunotherapy and the last dose was at least 28 days after the first administration of BT5528.

[0130] In some implementations, the patient must meet the following criteria: have metastatic or recurrent disease histologically confirmed as non-small cell lung cancer (NSCLC), ovarian cancer, triple-negative breast cancer (TNBC), gastric / upper gastrointestinal (GI) cancer, cancer, head and neck cancer, or urothelial carcinoma; have failed or are unsuitable for all appropriate treatment options according to local guidelines; and have evidence of radiographic progression in the most recent line of treatment.

[0131] In some embodiments, the patient has urothelial carcinoma and has previously received treatment with enfortumab vedotin (EV). In some embodiments, the patient experiences disease progression within 6 months of initiating EV treatment.

[0132] In some embodiments, the patient has adenocarcinoma subtype of NSCLC (adeno-NSCLC) and has exhausted all standard treatment options (including progression on or after platinum-based chemotherapy), and / or has experienced failure in at least one prior line of treatment and has evidence of radiographic progression in the most recent line of treatment. In some embodiments, the patient has EGFR, ALK, NTRK, ROS1, or other genomic tumor abnormalities. In some embodiments, the patient has not received appropriate treatment for the driver gene mutation. In some embodiments, the patient has not received immunotherapy for at least 28 days prior to the first administration of the pharmaceutical composition of the present invention. In some embodiments, the patient has not received chemotherapy for 14 days prior to the first administration of the pharmaceutical composition of the present invention. In some embodiments, the patient has not received anticancer therapy for 28 days or 5 half-lives (whichever is shorter) prior to the first administration of the pharmaceutical composition of the present invention. In some embodiments, the patient has a history of toxicity that has been resolved to grade 1 according to the Common Terminology Criteria for Adverse Events (CTCAE) version 5.0 (except for alopecia, which must not be higher than grade 2).

[0133] In some embodiments, the patient has not undergone experimental treatment within 4 weeks of the initial administration of the pharmaceutical composition of the present invention.

[0134] In some implementations, the patient is not currently being treated with strong inhibitors or inducers of CYP3A4 or strong inhibitors of P-gp (including herbal or food-based ones).

[0135] In some embodiments, the patient has no allergy to any component of the pharmaceutical composition of the present invention or to monomethylauratestine E (MMAE).

[0136] In some implementations, the patient does not have any significant medical condition, life-threatening disease, active uncontrolled infection, or organ system dysfunction (such as ascites, coagulopathy, or encephalopathy).

[0137] In some embodiments, the patient has not undergone any major surgery (excluding vascular access placement) within 4 weeks of the first administration of the pharmaceutical composition of the present invention.

[0138] In some embodiments, the patient has not received a live vaccine within 30 days of the first administration of the pharmaceutical composition of the present invention.

[0139] In some embodiments, the patient does not have uncontrolled, symptomatic brain metastases. In some embodiments, the patient has a stable neurological status after at least 4 weeks of local treatment without the use of steroids or at a stable or reduced dose of prednisone or an equivalent less than or equal to 10 mg daily.

[0140] In some implementations, the patient has no untreated CNS metastases. In some implementations, the patient has treated CNS metastases, and a) the CNS metastases have been clinically stable for at least 6 weeks prior to screening; b) if steroid treatment for the CNS metastases is required, the subject has received a stable dose or a reduced dose of prednisone or an equivalent at ≤20 mg / day for at least 2 weeks; c) baseline scans show no evidence of new or expanding brain metastases; d) the subject has no leptomeningeal disease; and e) the subject must not have any neurological dysfunctions that would confound neurological and other adverse events (AEs) assessments.

[0141] In some embodiments, the patient does not have uncontrolled hypertension (systolic blood pressure [BP] ≥139 mmHg; diastolic blood pressure ≥89 mmHg) prior to the first administration of the pharmaceutical composition of the present invention. In some embodiments, the patient has hypertension that has been stably controlled for at least 3 months prior to the first administration of the pharmaceutical composition of the present invention.

[0142] In some embodiments, the patient does not have uncontrolled hypertension (systolic blood pressure [BP] ≥160 mmHg; diastolic blood pressure ≥100 mmHg, unresponsive to intervention) at screening or prior to the first administration of the pharmaceutical composition of the present invention. In some embodiments, the patient has hypertension that has been stably controlled for at least 3 months prior to the first administration of the pharmaceutical composition of the present invention.

[0143] In some embodiments, the patient has no history of cerebrovascular events (stroke or transient ischemic attack), unstable angina, myocardial infarction, congestive heart failure, or New York Heart Association Class III-IV symptoms within 6 months prior to the first administration of the pharmaceutical composition of the present invention.

[0144] In some embodiments, the patient does not have a mean resting corrected QT interval (QTcF) >470 msec within 6 months prior to the first administration of the pharmaceutical composition of the present invention.

[0145] In some embodiments, within 6 months prior to the first administration of the pharmaceutical composition of the present invention, the patient has no factors that increase the risk of QTc prolongation or arrhythmic events, such as heart failure, hypokalemia, congenital long QT syndrome, a family history of unexplained sudden death under the age of 40, or any concomitant medications known to prolong the QT interval.

[0146] In some embodiments, the patient does not have any clinically significant abnormalities in the rhythm, conduction, or morphology of a resting electrocardiogram (ECG) within 6 months prior to the first administration of the pharmaceutical composition of the present invention, such as complete left bundle branch block or third-degree heart block.

[0147] In some implementations, the patient does not have human immunodeficiency virus (HIV) or acquired immunodeficiency syndrome (AIDS).

[0148] In some implementations, the patient has well-controlled HIV and meets all of the following inclusion criteria: a. CD4+ count ≥350 cells / μL; b. HIV viral load <400 copies / mL; c. No history of opportunistic infections in the past 12 months; d. Has been receiving stable antiretroviral therapy (ART) for at least four weeks. The use of ART should be discussed with a medical monitor on a case-by-case basis.

[0149] In some implementations, the patient does not have positive hepatitis B surface antigen and / or anti-hepatitis B core antibody.

[0150] In some implementations, the patient has a negative polymerase chain reaction (PCR) test and has appropriate antiviral therapy.

[0151] In some implementations, if a patient tests positive for hepatitis C virus (HCV) antibodies, the patient is considered to have an active hepatitis C infection with a positive viral load.

[0152] In some implementations, the patient has received treatment for hepatitis C infection and has a sustained virological response for ≥12 weeks.

[0153] In some embodiments, the patient is free of thromboembolic events and / or bleeding disorders (e.g., deep vein thrombosis [DVT] or pulmonary embolism [PE]) within 3 months prior to the first administration of the pharmaceutical composition of the present invention.

[0154] In some embodiments, the patient does not have another malignancy within 3 years prior to the first administration of the pharmaceutical composition of the present invention. In some embodiments, the patient does not have any residual lesions of a previously diagnosed malignancy (excluding basal cell carcinoma, squamous cell carcinoma of the skin, cervical intraepithelial neoplasia / cervical carcinoma in situ, melanoma in situ, or ductal carcinoma in situ of the breast that have received adequate treatment for a curative purpose).

[0155] In some embodiments, the patient had not received systemic anti-infective treatment and had no fever during the last 14 days prior to the first administration of the pharmaceutical composition of the present invention.

[0156] Example

[0157] Example 1. Phase I / II study of the safety, pharmacokinetics, and preliminary clinical activity of BT5528 in patients with advanced malignancies associated with EphA2 expression.

[0158] 2.1 Objectives

[0159] Main objectives

[0160] The primary objectives of the dose escalation phase (parts A-1 and A-2) are: Evaluate the safety and tolerability of BT5528 as monotherapy (Part A-1) and in combination with nivolumab (Part A-2), administered weekly, in patients with advanced solid tumors associated with EphA2 expression and / or specific tumors identified as positive for EphA2 expression. Determine the MTD (if observed) of BT5528 as monotherapy (Part A-1) and in combination with nivolumab (Part A-2), and determine one or more recommended phase 2 doses (RP2D).

[0161] The main objectives of the expansion phase (Part B) are: The clinical activity of BT5528 as monotherapy in patients with solid tumors previously known to have high EphA2 expression was evaluated in one or more R2PD dosing regimens (cohort B-1: urothelial carcinoma, cohort B-2: ovarian cancer, cohort B-3: NSCLC, cohort B-4: head and neck, cohort B-5: TNBC, and cohort B-6: stomach / upper GI).

[0162] Secondary objectives

[0163] The secondary objective of the dose escalation phase (parts A-1 and A-2) in this study was: To evaluate preliminary signals of antitumor activity obtained by administering BT5528 as monotherapy (Part A-1) and in combination with nivolumab (Part A-2) in patients with advanced solid tumors associated with EphA2 expression and / or specific tumors identified as positive for EphA2 expression. The secondary objective of the extension phase (Part B) study was: To evaluate the safety and tolerability of BT5528 as a monotherapy in patients with tumors known to have high EphA2 expression, and to investigate whether tumor EphA2 expression levels can predict the clinical activity of BT5528 monotherapy in patients with tumors known to have high EphA2 expression.

[0164] The secondary objective of all parts of the study is: Determine the pharmacokinetic (PK) parameters of BT5528 and MMAE (if applicable); Determine the incidence of anti-drug antibodies (ADA).

[0165] 2.2. Research Design

[0166] This study is a Phase I / II, first-in-human, open-label dose-escalation study of BT5528 as monotherapy (Parts A-1 and B) and in combination with nivolumab (Part A-2). The study has two parts: Part A, dose escalation; and Part B, dose extension.

[0167] 2.3. Study the drug, dosage, and method of administration: In Part A, BT5528 was administered via intravenous infusion over 1 hour in escalating doses; in Part B, it was administered according to the RP2D regimen. In Part A, 480 mg of nivolumab was administered intravenously every 4 weeks, or 240 mg of nivolumab every 2 weeks.

[0168] 2.4. Inclusion Criteria – All Patients: Patients must meet the following criteria to be included in the study: 1. The patient or legal guardian shall sign and date a written informed consent form in accordance with local guidelines before any specific research procedure, sampling or analysis is conducted.

[0169] If a patient refuses to participate in any voluntary part of this study (e.g., tumor biopsy), he / she will not be penalized or lose any benefit, nor will he / she be excluded from other aspects of this study.

[0170] 2. Must be at least 18 years old when signing the informed consent form.

[0171] 3. Eastern Cooperative Oncology Group (ECOG) performance status score of 0 or 1.

[0172] 4. According to the Remission Evaluation Criteria in Solid Tumors (RECIST) version 1.1, the patient must have a measurable disease.

[0173] 5. Acceptable organ function, as demonstrated by the following laboratory data: - Renal function, as follows: creatinine clearance ≥50 mL / min, calculated by the Cockcroft-Gault formula or as measured by 24-hour urine collection.

[0174] - Total bilirubin ≤ 1.5 × ULN (upper limit of normal)

[0175] - Serum albumin ≥2.5 g / dL

[0176] - In the presence of liver metastases, aspartate aminotransferase (AST) ≤2.5 × ULN or ≤5 × ULN

[0177] - In the presence of liver metastases, alanine aminotransferase (ALT) ≤2.5 × ULN or ≤5 × ULN

[0178] - International Normalized Ratio (INR) <1.3 or ≤ Institutional ULN

[0179] 6. Acceptable hematological function (no red blood cell or platelet transfusions within 4 weeks of the first administration of BT5528, and no use of growth factors):

[0180] - Hemoglobin ≥ 9 g / dL

[0181] - Absolute neutrophil count (ANC) ≥ 1500 cells / mm 3

[0182] - Platelet count ≥75,000 cells / mm 3

[0183] 7. Women of childbearing potential (WOCBP) must have a negative pregnancy test (negative serum test at screening and negative urine or serum test within 3 days prior to the first administration of BT5528). Male patients with female partners of childbearing potential and female patients of childbearing potential must adhere to highly effective contraception (oral and hormonal contraceptives are permitted) during their participation in the study and for 6 months after the last administration of BT5528 (BT5528 IB), or for patients continuing nivolumab-only treatment after discontinuation of BT5528 treatment for 5 months after the last administration of nivolumab (nivolumab USPI or SmPC). This highly effective contraception must be at least as conservative as the CTFG-recommended failure rate of less than 1%. Male patients must also avoid sperm donation during their participation in the study and for 5 or 6 months after the last dose of nivolumab or BT5528, and women must not breastfeed or donate eggs during this period.

[0184] 8. All patients must have tumor tissue (fresh or archived) available for analysis of EphA2 tumor expression and other biomarkers. In cases where tumor tissue is unavailable, the patient must be willing to undergo a biopsy to provide a fresh tumor sample. Specifications regarding tissue requirements are provided in the laboratory manual.

[0185] 9. According to the researchers' assessment, the life expectancy after the start of BT5528 treatment is ≥12 weeks.

[0186] 10. Must be willing and able to comply with the research plan and research procedures.

[0187] Additional inclusion criteria – Part A only

[0188] 11. Patients with histologically confirmed metastatic or recurrent malignant solid tumors who have exhausted all appropriate treatment options according to local guidelines and must have experienced failure in at least one prior line of treatment, with evidence of radiographic progression in the most recent line of treatment. If applicable, for patients who have received prior immunotherapy, the last dose must have been administered at least 28 days prior to the first administration of BT5528 study treatment.

[0189] Pancreatic cancer patients are permitted to be enrolled, but less than 50% of the total patients are planned to be enrolled in each dose escalation cohort. If the sponsor or SRC deems it necessary to enrich biomarkers, safety, or PK assessments in a specific tumor type, it may decide at any time during dose escalation to request recruitment of a specific tumor type or subtype.

[0190] 12. Patients enrolled in cohorts A1-8 or subsequent cohorts of monotherapy in part A-1, or cohorts A2-3 or subsequent cohorts of combination therapy in part A-2, must have tumor tissue (fresh or archived) available for prospective or retrospective analysis of EphA2 tumor expression by the central laboratory, but may be enrolled without prior confirmation of EphA2 expression.

[0191] Additional inclusion criteria – Part B: Single-drug treatment cohort

[0192] 13. Patients with histologically confirmed metastatic or recurrent non-small cell lung cancer (NSCLC), ovarian cancer, triple-negative breast cancer (TNBC), gastric / upper gastrointestinal (GI) cancer, cancer, head and neck cancer, or urothelial carcinoma meet the criteria; and must have failed or be ineligible for all appropriate treatment regimens according to local guidelines; and must have evidence of recent radiographic progression of their treatment line.

[0193] 14. Patients with urothelial carcinoma who have previously received emfumab-vitoltin (EV) therapy are eligible to participate in this study. Patients who received EV and experienced disease progression within 6 months of treatment initiation are planned to be enrolled at less than 50% of the total enrolled cohort.

[0194] 2.5. Exclusion criteria – All patients: Patients meeting any of the following criteria will be excluded from the study: 1. Chemotherapy was administered within 14 days prior to the first administration of study treatment. For other anticancer therapies, treatment was administered within 28 days or 5 half-lives, whichever is shorter. For immunotherapy, including immune checkpoint inhibitors, treatment was administered within 28 days prior to the first administration of study treatment.

[0195] 2. An experimental treatment was administered within 4 weeks following the initial study treatment with BT5528.

[0196] 3. According to the Common Terminology Standard for Adverse Events (CTCAE) version 5.0, prior toxicity must be mitigated to Grade 1 (except for Grade 2 alopecia, which can be mitigated).

[0197] 4. Currently, treatment is being carried out using strong inhibitors or inducers of CYP3A4 or strong inhibitors of P-gp (including herbal or food-based inhibitors).

[0198] 5. Known sensitivity to the investigational drug or any component of monomethylauresstatin E (MMAE).

[0199] 6. Major medical conditions, life-threatening illnesses, uncontrolled active infections or organ system dysfunctions (such as ascites, coagulopathy, encephalopathy), or other reasons that the investigator believes may jeopardize patient safety or interfere with or impair the integrity of the study results, including consideration of gastrointestinal, skin and pulmonary comorbidities, and including review of chest CT screenings to ensure that there are no clinically significant comorbidities.

[0200] 7. Major surgery (excluding vascular access placement) must be performed within 4 weeks of the initial BT5528 study treatment, and the patient must have fully recovered before starting study treatment.

[0201] 8. Administer the live vaccine within 30 days of starting the study treatment.

[0202] 9. Untreated CNS metastases. Subjects with treated CNS metastases are eligible to participate in this study if all of the following criteria are met: a. The CNS metastases had been clinically stable for at least 6 weeks prior to screening. b. If CNS metastases require steroid therapy, the subject should receive a stable or tapering dose of prednisone or an equivalent at ≤20 mg / day for at least 2 weeks. c. Baseline scan showed no evidence of new or expanded brain metastases. d. The subject did not have leptomeningeal disease. e. Subjects must not have any neurological dysfunctions that would confuse neurological and other adverse event assessments.

[0203] 10. Uncontrolled hypertension (systolic blood pressure ≥160 mmHg or diastolic blood pressure ≥100 mmHg, unresponsive to intervention) at screening or before starting the study drug.

[0204] 11. Any historical or current evidence of disease, treatment, or laboratory abnormalities that may confound research results, interfere with patient participation, or indicate that participation in the study based on the investigator's opinion is not in the best interests of the patient, including but not limited to: a. Patients with a history of a cerebrovascular event (stroke or transient ischemic attack), unstable angina, myocardial infarction, congestive heart failure, or New York Heart Association class III-IV symptoms recorded within 6 months prior to the first administration of BT5528, or the following conditions: i. Mean resting corrected QT interval (QTcF) > 470 msec ii. Any factor that increases the risk of QTc prolongation or arrhythmic events, such as heart failure, hypokalemia, congenital long QT syndrome, a family history of unexplained sudden death in individuals under 40 years of age, or any concomitant medication known to prolong the QT interval. iii. Any clinically significant abnormalities in rhythm, conduction, or morphology on the resting electrocardiogram (ECG) (assessed by the investigator), such as complete left bundle branch block or third-degree atrioventricular block. 12. Known human immunodeficiency virus (HIV) or acquired immunodeficiency syndrome (AIDS) Note: Patients with well-controlled HIV infection are allowed to be included if they meet all of the following criteria at the time of enrollment: a) CD4+ T cell (CD4+) count ≥350 cells / uL; b) HIV viral load <400 copies / mL; c) No history of opportunistic infections in the past 12 months; d) Has been receiving stable antiretroviral therapy (ART) for at least 4 weeks. The use of antiretroviral therapy is permitted, but should be discussed with a medical monitor on a case-by-case basis.

[0205] 13. Patients who are positive for hepatitis B surface antigen and / or anti-hepatitis B core antibody. Patients with negative polymerase chain reaction (PCR) tests are permitted to receive appropriate antiviral therapy.

[0206] 14. If hepatitis C virus (HCV) antibody is positive, then active hepatitis C infection with a positive viral load is considered (if antibody is negative, viral load is not applicable). Patients who have received treatment for hepatitis C infection and have a documented sustained virological response for ≥12 weeks may be included.

[0207] 15. A thromboembolic event and / or hemorrhagic condition (e.g., deep vein thrombosis [DVT] or pulmonary embolism [PE]) occurred within 3 months prior to the first administration of BT5528 in the study.

[0208] 16. History of pneumonia with residual symptoms.

[0209] 17. A history of another malignancy within 3 years prior to the first administration of BT5528, or any evidence of residual disease from a previously diagnosed malignancy (excluding basal cell carcinoma, squamous cell carcinoma of the skin, cervical intraepithelial neoplasia / cervical carcinoma in situ or melanoma in situ or ductal carcinoma in situ of the breast that have been adequately treated for a curative purpose).

[0210] 18. Patients who have received systemic anti-infective therapy or have fever within 14 days prior to the first administration of BT5528 study treatment.

[0211] 19. Psychological, family, social or geographical conditions that preclude compliance with the protocol and / or the follow-up procedures outlined in the protocol.

[0212] Additional exclusion criteria A-2: Nivolumab combination cohort

[0213] 20. Previous intolerance to immune checkpoint inhibitors

[0214] 21. Hypersensitivity reactions to checkpoint inhibitor therapy are known.

[0215] 22. Previous organ transplants (including allogeneic transplants)

[0216] 23. Diagnosis of clinically relevant immunodeficiency

[0217] 24. Active systemic infection requiring treatment

[0218] 25. Daily doses exceeding 10 mg of prednisone equivalent or other potent immunosuppressants.

[0219] 26. History of autoimmune diseases other than hair loss or vitiligo

[0220] 2.6 Correlation Test: All patients will be required to provide archived tumor material or fresh tumor biopsy to assess EphA2 expression levels and additional molecular genetic characteristics (i.e., assessment of specific somatic mutations, etc.). This material should be provided in the form of tissue blocks or 10–15 paraffin-impregnated unstained slides.

[0221] For cohorts A-1 receiving monotherapy or A-2 receiving combination therapy, or cohorts A2-3 or subsequent cohorts, tumor tissue (fresh or archived) must be submitted to the central laboratory for prospective or retrospective analysis of EphA2 tumor expression, but inclusion is permitted without prior confirmation of EphA2 expression. When a single tumor type or subtype has demonstrated indications of efficacy, the SRC may review the requirement for EphA2 expression before initiating treatment.

[0222] For Part B (all expanded cohorts), EphA2 expression will initially be analyzed retrospectively, and confirmation of EphA2 expression is not required prior to the study of treatment. However, if efficacy analyses for each individual indication or cohort indicate the need to enrich patients with EphA2 expression in one or more specific tumor indication cohorts, confirmation of EphA2 expression in these cohorts will be required before initiating the study of treatment.

[0223] To investigate the intratumoral PK / pharmacodynamic effects of BT5528, tumor biopsies will be collected post-dose, subject to patient consent. Post-dose tumor biopsies are optional for all patients. They will be collected within 4 hours to 7 days after the end of any BT5528 infusion (EOI) during Cycle 1, or in any subsequent cycle if not feasible. Refer to the Evaluation Plan (SoA) for further details.

[0224] Blood samples will also be collected before and after administration to assess pharmacodynamics, response, and treatment resistance biomarkers, such as somatic mutations in circulating tumor DNA (ctDNA), ADA, and pharmacogenomics analysis.

[0225] 2.7. Statistical Methods: Dose escalation (applicable to A-1 and A-2 respectively): The actual number of dose levels to be investigated in this study will depend on the determination of intolerable doses based on dose-limiting toxicity (DLT). MTD will be defined according to DLT (see Section 5). Other safety data, as well as PK characterization profiles and any trends in antitumor activity observed during the study, will be used to determine RP2D, which is not greater than MTD (or, in the absence of a stated MTD, not greater than the maximum administered dose [MAD], which is defined as the highest dose in each escalation study).

[0226] The first two dose levels will use a 3+3 design. At least three evaluable patients will be recruited for each dose level and evaluated for 28 days before escalating to the next dose level. After confirming tolerability at dose level 1, the dose will be allowed to escalate to dose level 2 in increments not exceeding 100%. Treatment cycles will occur consecutively according to the SoA (Socially Assigned) protocol. If one patient experiences DLT (Disease-Related Lung Disease), three other patients will receive the same dose. Before proceeding to the next dose level, a cohort of at least three patients completing one treatment cycle (28 days) will be evaluated. See Section 5 for further details.

[0227] Based on the tolerability evidence from the first two dose levels, all subsequent dose escalations will be based on a Continuous Reassessment (CRM) approach. This approach uses a two-parameter Bayesian logistic regression (BLRM) model and the overdose control (EWOC) principle. The next higher dose escalation level will be the dose with the highest posterior probability of a DLT occurring within the target range (20%, 33%) that meets the overdose criteria (a dose level with a probability of being found unsafe ≤25% (DLT rate ≥33%)). BLRM will be applied to the accumulated DLT / safety data, and the results will be provided to the SRC. After reviewing this data, the SRC will make recommendations on precise dose escalation.

[0228] Each cohort for each RP2D in Part B will employ a Simon 2-stage design with p0 = 0.11, p1 = 0.30, one-sided α = 0.05, and power of 80%, where p0 and p1 are the null and surrogate hypotheses for overall response rate (ORR). If three or more patients out of the initially recruited 14 show OR, the sponsor will consider whether the cohort is suitable for Phase 2, recruiting an additional 26 patients; otherwise, the cohort will be terminated. The additional 26 patients will be included in at most two Part B cohorts.

[0229] The efficacy results from Part A of this study can be used to guide enrichment of specific cohorts based on EphA2 status. For all cases in Part B of this study, efficacy will be determined based on OR.

[0230] Due to these changes, the maximum number of patients recruited for this study was 288: 72 patients were included in Part A-1, 24 patients in Part A-2, and a maximum of 192 patients in Part B cohort, provided that all indications for all tests met the clinical efficacy criteria according to the Simon Phase 2 analysis described above.

[0231] For ORR and CBR, estimates and associated 95% CIs will be calculated (based on the Clopper-Pearson method). For PFS and OS, Kaplan-Meier curves will be generated, and the median time of occurrence and associated 95% CIs will be provided.

[0232] Example 3. Core Results at the End of Phase I Dose Escalation

[0233] BT5528 demonstrated significant antitumor activity in advanced solid tumors with a favorable overall safety / tolerability profile. The antitumor activity of BT5528 was tested in heavily pre-treated cancer patients. A 67% ORR was observed in patients with urothelial carcinoma having tumors associated with EphA2 expression. Figure 2 A correlation began to emerge between antitumor activity and EphA2 expression. Figure 3 A). Bt5528 also showed a reduction in tumor volume in a female patient with urothelial carcinoma who had previously received four lines of therapy. Figure 4 The emerging safety features of BT5528 distinguish it from other EphA2-targeting molecules: - No IMP-related clinical bleeding / coagulopathy - No evidence of IMP-related ocular toxicity or clinically significant skin toxicity. Part B had no reports of ILD / pneumonia. - No significant hepatotoxicity Gastrointestinal and hematological toxicities are dose-dependent and controllable. Improving the incidence / severity of CINV through antiemetic preventive measures The use of growth factors is uncommon. -BP / Vital Signs: No trend observed -ECG: No trend of concern regarding QT issues Example 4. Phase I / II Results of the Combination Phase II data were also collected from heavily treated patients after the completion of Phase I dose escalation. These data confirmed the antitumor and safety / tolerability signals identified in Example 3.

[0234] The Phase II data was merged with the existing Phase I data, including the merged demographic data. Figure 7A ) and solid tumor type classification ( Figure 7B The security features of BT5528 remain promising. Figure 8A –BT5528 Security Overview; Figure 8B –The most common treatment-related adverse events (≥15%)–RP2D is well-tolerated; Figure 8C – Treatment-related adverse events of concern – occurred at low rates at all doses.

[0235] This provides promising early response data for urothelial carcinoma. Figure 9A - Tumor remission waterfall chart; Figure 9B –Tumor response spider diagram). The prior treatment history of 7 patients with urothelial carcinoma who received intensive pre-treatment is as follows: Previous treatment of seven UC patients in remission:

[0236] All patients had previously received immunotherapy. Two patients received ADCs, one of whom had previously received emfumab-vitoltin (EV).

[0237] The efficacy evaluable group was defined as all enrolled patients who had measurable disease at baseline, had received at least one dose of BT5528, and underwent at least one adequate post-baseline disease assessment. Four patients were excluded due to lack of post-baseline assessment. A fifth patient was excluded from the waterfall plot because target lesion data were not evaluable in a single post-baseline assessment.

[0238] Overall, among the 18 patients, 7 (6 confirmed) showed partial remission, and 3 (1 confirmed) had stable disease for more than 4 months. 8 patients showed progressive disease. The clinical benefit rate (CBR) (confirmed partial remission or stable disease) and objective response rate (ORR) (partial remission) were 39%. Specifically, for patients receiving 6.5 mg / m² every two weeks... 2 Of the 16 patients who underwent BT5528 treatment, 5 (4 confirmed) showed partial remission, and 3 (1 confirmed) had stable disease for more than 4 months. The CBR and ORR were 31%. When the B-1 extended cohort (prophylactic antiemetic 6.5 mg / m²) was used... 2 When Q2W was separated, an ORR of 45.5% was observed in patients with evaluable efficacy (n = 11).

[0239] In patients with other tumors (including ovarian cancer) Figures 9C-9D Tumor shrinkage (including formal remission) was also observed in patients with head and neck cancer (Fig. 9E) and gastric / upper gastrointestinal cancer (Fig. 9F). Stable disease was also observed in patients with NSCLC (Fig. 9G) and TNBC (Fig. 9H). Response data for urothelial carcinoma and ovarian cancer are summarized. Figure 9I ).

[0240] Example 5. BT5528 dose modeling.

[0241] 5 mg / m² per week for BT5528 2 Dosing was performed, and the PK characteristics of MMAE were simulated. As simulated in a typical subject with a BSA of 1.8 m² using the PK model as of the data cutoff date, this is expected to provide a 54% higher dose / exposure ratio for MMAE compared to a dose of 6.5 mg / m² administered every two weeks. Figures 10A-10B ) ≥3 levels of TRAE were observed. Figure 11A Dosage adjustments due to treatment-related adverse events (AEs) Figure 11B ) and grade ≥3 neutropenia ( Figure 11C ) and MMAE C maxAn exposure-safety relationship exists. The exposure-mitigation analysis is based on available data up to the data cutoff. Logistic regression was used to test the exposure-mitigation relationship. The simulated adverse event incidence was derived from the virtual patient population from MMAE cycle 1 C. max Driven by logistic regression. In summary, these data indicate that 5 mg / m 2 A weekly dosing regimen could be a promising dose for further investigation and could provide greater efficacy while maintaining an acceptable safety / tolerability profile.

[0242] Example 6: BT5528 Dosage Optimization

[0243] Based on the dose modeling work described in this paper, further dose optimization studies were conducted to investigate the 5 mg / m² dose of BT5528. 2 Safety / tolerability and efficacy of weekly doses. Patients in this cohort received prophylactic antiemetics.

[0244] Interim safety / tolerance data such as Figures 12A-12C As shown. The safety analysis population included patients who had received at least one dose of BT5528. These data indicate that BT5528 at 5 mg / m² is safe. 2 It was well tolerated at the QW dose, and 5 mg / m² 2 QW dosage showed a correlation with 6.5 mg / m 2 The Q2W dose has comparable tolerability. Figure 12C Including BT5528 at 5 mg / m 2 QW dose and 6.5 mg / m 2 An overview of relevant TEAEs at the Q2W dose, where the TEAE data cutoff for 6.5 mg / m² is more recent than in the previous examples.

[0245] The study also investigated 5 mg / m 2 Efficacy of QW dose. The evaluable population includes all patients who have received at least one dose of BT5528 and have undergone at least one post-baseline assessment. For ORR (CR+PR) and CBR (CR+PR+SD ≥ 4 months from C1D1), patients without post-baseline tumor assessment will be classified as non-evaluable (NE) and considered as non-responders.

[0246] As of the data cutoff date, 3 patients (27.3%) showed partial remission, and 4 patients (36.4%) had stable disease for more than 4 months. Figure 13A Patient remission is graphically represented as a spider diagram. Figure 13B ) and waterfall picture ( Figure 13CAs of the data cutoff date, the study is still ongoing: three patients are still receiving BT5528, including two patients with unconfirmed partial remission and one patient with stable disease.

[0247] Based on this, 5 mg / m 2 The QW dosing regimen has demonstrated acceptable safety and tolerability, as well as indications of efficacy in patients with urothelial carcinoma.

Claims

1. A method of treating a human patient, the method comprising administering BT5528 or a pharmaceutical composition comprising BT5528 to the patient.

2. A method of treating a solid tumor in a human patient, the method comprising administering BT5528 or a pharmaceutical composition comprising BT5528.

3. A method for maintaining or reducing the volume of a solid tumor in a patient, the method comprising administering BT5528 or a pharmaceutical composition containing BT5528.

4. The method of claim 2 or 3, wherein the solid tumor is an advanced malignant tumor.

5. The method of any one of claims 2 to 4, wherein the solid tumor or advanced malignant tumor is associated with EphA2 expression in a human patient.

6. The method of any one of claims 1 to 5, said method comprising administering approximately 4-8 mg / m² every two weeks. 2 The dosage, optionally about 5-7 mg / m² 2 The dosage, optionally approximately 6.5 mg / m² 2 The patient was given a dose of BT5528 or a pharmaceutical composition containing BT5528.

7. The method of any one of claims 1 to 5, said method comprising administering approximately 3-7 mg / m² weekly. 2 The dosage, optionally about 4-6 mg / m² 2 The dosage, optionally about 5 mg / m 2 The patient was given a dose of BT5528 or a pharmaceutical composition containing BT5528.

8. The method of any one of claims 2-7, wherein the solid tumor or advanced malignant tumor is selected from the group consisting of: lung cancer (e.g., NSCLC), ovarian cancer, fallopian tube cancer, breast cancer (e.g., TNBC), head and neck cancer, gastric / upper gastrointestinal (GI) cancer, pancreatic cancer, urothelial carcinoma (e.g., bladder cancer, urethral cancer, urethral bladder cancer, and urothelial carcinoma), bone cancer, rectal cancer, gastric cancer, and squamous cell carcinoma of unknown origin.

9. The method of any one of claims 1 to 8, further comprising administering an anti-PD-1 antibody or an anti-PD-L1 antibody to the patient, optionally wherein the anti-PD-1 antibody is nivolumab.

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