EphA2-specific bicyclic peptide ligand and its use

Abstract

This invention relates to the bicyclic toxin complex BT5528 or its pharmaceutically acceptable salts, or its pharmaceutical composition, and its use.

Description

[Technical Field]

[0001] The present invention relates to a bicyclic toxin complex, a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. The present invention also provides the use of a bicyclic toxin complex, a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof for the prevention or treatment of a disease, disorder, or condition characterized by overexpression of EphA2 in affected tissue. [Background technology]

[0002] Cyclic peptides can bind to protein targets with high affinity and target specificity, and are therefore an attractive molecular class for therapeutic drug development. In fact, several cyclic peptides have already been successfully used clinically, for example, as the antimicrobial peptide vancomycin, the immunosuppressant cyclosporine, or the anticancer drug octreotide (Driggers et al. (2008), Nat Rev Drug Discov 7 (7), 608-24). Good binding properties result from the relatively large interaction surface formed between the peptide and the target, and the reduced conformational flexibility of the cyclic structure. Typically, macrocyclic molecules bind to a surface of several hundred square angstroms, for example, the cyclic peptide CXCR4 antagonist CVX15 (400 Å). 2 (Wu et al. (2007), Science 330, 1066-71), a cyclic peptide having an Arg-Gly-Asp motif that binds to integrin αVb3 (355Å 2 )(Xiong et al. (2002), Science 296 (5565), 151-5) or upain-1 (603Å), a cyclic peptide inhibitor that binds to urokinase-type plasminogen activator. 2 ;Zhao et al. (2007), J Struct Biol 160 (1), 1-10).

[0003] Due to their cyclic configurations, macrocyclic peptide molecules are less flexible than linear peptides, resulting in less entropy loss during binding to the target and consequently higher binding affinity. This reduced flexibility also leads to the fixation of target-specific conformations, increasing binding specificity compared to linear peptides. This effect is exemplified by the potent and selective inhibitory effect of matrix metalloproteinase 8 (MMP-8), which loses more selectivity than other MMPs when its ring is opened (Cherney et al. (1998), J Med Chem 41 (11), 1749-51). The favorable binding properties achieved by macrocyclization are even more pronounced in polycyclic peptides having one or more peptide rings, such as vancomycin, nisin, and actinomycin.

[0004] Various research teams have previously linked polypeptides containing cysteine ​​residues to synthetic molecular structures (Kemp and McNamara (1985), J. Org. Chem; Timmerman et al. (2005), ChemBioChem). Meloen and collaborators used tris(bromomethyl)benzene and related molecules for the rapid and quantitative cyclization of multiple peptide loops onto a synthetic scaffold to mimic the structure of a protein surface (Timmerman et al. (2005), ChemBioChem). A method for generating candidate drug compounds is described, which are produced by linking a cysteine-containing polypeptide to a molecular scaffold such as TATA(1,1',1''-(1,3,5-triazinan-1,3,5-triyl)tripropa-2-en-1-one, Heinis et al. Angew Chem, Int Ed. 2014; 53:1602-1606).

[0005] A phage display-based combinatorial approach has been developed to construct and screen large libraries of bicyclic peptides against targets of interest (Heinis et al. (2009), Nat Chem Biol 5 (7), 502-7 and WO2009 / 098450). Briefly, a combinatorial library of linear peptides containing two regions (Cys-(Xaa)6-Cys-(Xaa)6-Cys) of three cysteine ​​residues and six random amino acids is displayed on a phage and cyclized by covalently bonding the cysteine ​​side chain to a small molecule scaffold. [Overview of the Initiative]

[0006] According to one embodiment, the present invention provides a method for treating a human patient, comprising administering BT5528 or a pharmaceutical composition comprising BT5528.

[0007] In another embodiment, the present invention provides a method for treating solid tumors in human patients, comprising administering BT5528 or a pharmaceutical composition comprising BT5528. The present invention also provides BT5528 or a pharmaceutical composition comprising BT5528 for use in a method for treating solid tumors in human patients. [Modes for carrying out the invention]

[0008] 1. Compounds and definitions: As used herein, the term "BT5528" refers to a bicyclic toxin complex having the structure shown below, the molecular scaffold being 1,1',1''-(1,3,5-triazinan-1,3,5-triyl)tripropa-2-en-1-one (TATA), and the peptide ligand having the amino acid sequence: (β-Ala)-Sar 10 -A(HArg)DC i (HyP)LVNPLC ii LHP(D-Asp)W(HArg)C iii (Sequence ID 1) This includes, where Sar is sarcosine, HArg is homoarginine, and HyP is hydroxyproline.

[0009] As used herein, the term “pharmaceutically acceptable salt” refers to a salt that, within the bounds of sound medical judgment, is suitable for use in contact with human and lower animal tissues without excessive toxicity, irritation, or allergic reactions, and that is commensurate with a reasonable benefit / risk ratio. pharmaceutically acceptable salts are well known in the art. For example, S.M. Berge et al. describe pharmaceutically acceptable salts in detail 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 salts of amino groups 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 by using other methods used in the art, such as ion exchange. Other pharmaceutically acceptable salts include adipine, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphor sulfonate, citrate, cyclopentanepropionate, digluconate, dodecyl sulfate, ethanesulfonate, formate, fumarate, glucoheptone, glycerophosphate, gluconate, hemiphosphate, heptaneate, hexanoate, hydroiodide, and 2-hydroxyethanes. Examples include ruhonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, and valerate. [Chemical]

[0010] Salts derived from suitable bases include alkali metal salts, alkaline earth metal salts, ammonium salts, N + (C 1-4 (alkyl)4 salts. Representative alkali metal salts or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like. Further pharmaceutically acceptable salts include, where appropriate, non-toxic ammonium, quaternary ammonium, and amine cations formed using counterions such as halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, lower alkyl sulfonates, and aryl sulfonates. The salt forms are within the scope of the present invention, and reference to a peptide ligand is understood to include the salt forms of the ligand.

[0011] The salts of the present invention can be synthesized from a parent compound containing a basic or acidic moiety by conventional chemical methods such as those described in Pharmaceutical Salts: Properties, Selection, and Use, P. Heinrich Stahl (Editor), Camille G. Wermuth (Editor), ISBN: 3-90639-026-8, Hardcover, 388 pages, August 2002. Generally, such salts can be prepared by reacting the free acid or base form of these compounds with a suitable base or acid in water, in an organic solvent, or in a mixture of both.

[0012] As used herein, the term "about" is intended to have a meaning within 10% of a given value or range. In some embodiments, the term "about" refers to within 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, or 1% of a particular value.

[0013] Unless otherwise specified, the structures shown herein also mean all isomers (e.g., enantiomers, diastereomers, and geometric (or conformational) forms) of that structure; for example, R and S configurations for each chiral center, Z and E double bond isomers, and Z and E conformational isomers. Thus, single stereochemical isomers of the compounds of the present invention, as well as mixtures of enantiomers, diastereomers, and geometric (or conformational) forms, are within the scope of the present invention. Unless otherwise specified, all tautomeric forms of the compounds of the present invention are within the scope of the present invention. Furthermore, unless otherwise specified, the structures shown herein also mean that the compounds differ only in the presence of one or more isotopically enriched atoms; for example, the substitution of hydrogen with deuterium or tritium, or 13 C- or 14 Compounds having this structure, including carbon substitution with 1C-enriched carbon, are within the scope of the present invention. Such compounds are useful, for example, as analytical tools, as probes in biological assays, or as therapeutic agents according to the present invention.

[0014] 2. Treatment Method In one embodiment, the present invention provides a method or use for treating solid tumors in human patients. In some embodiments, the solid tumor is a progressive malignant tumor, and the method involves administering BT5528 or a pharmaceutical composition containing BT5528 to the patient. In some embodiments, the solid tumor or progressive tumor is associated with EphA2 expression.

[0015] 3. Combination therapy 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 treating solid tumors. In some embodiments, the treatment method of the present invention comprises the administration of BT5528 or a pharmaceutical composition comprising BT5528 and further administration of an anti-PD-1 antibody or an anti-PD-L1 antibody to a human patient.

[0016] In some embodiments, the anti-PD-1 antibody is selected from the list consisting of pembrolizumab, nivolumab, semiprimab, dostallimab, letifanlimab, voplaterimab, spartalizumab, camrelizumab, cintilimab, tislerizumab, tripalimab, INCMGA00012, AMP-224, AMP-514, and acrixolimab.

[0017] In some embodiments, the anti-PD-L1 antibody is selected from a list consisting of atezolizumab, avelumab, durvalumab, KN035, and cosivelimab.

[0018] In some embodiments, the method of the present invention comprises administering BT5528 or a pharmaceutical composition containing BT5528, and further administering nivolumab to a human patient.

[0019] Nivolumab may be administered as described on the label, which can be found at https: / / www.opdivohcp.com / dosing / dosing-schedules, and this content is incorporated herein by full attribution. In some embodiments, nivolumab is administered at a dose of 240 mg every two weeks. In some embodiments, nivolumab is administered at a dose of 480 mg every four weeks. In some embodiments, nivolumab is administered as a 30-minute intravenous infusion.

[0020] In some embodiments of the combination use, the patient is not previously known to be intolerant to immune checkpoint inhibitors. In some embodiments of the combination use, the patient is not known to be hypersensitive to checkpoint inhibitor treatment. In some embodiments of the combination use, the patient has no history of organ transplantation. In some embodiments of the combination use, the patient has not been previously diagnosed with clinically relevant immunodeficiency. In some embodiments of the combination use, the patient does not have an active systemic infection requiring treatment. In some embodiments of the combination use, the patient is not taking more than 10 mg of prednisone equivalent or other potent immunosuppressants daily. In some embodiments of the combination use, the patient has no history of autoimmune diseases other than alopecia or vitiligo. In some embodiments of the combination use, the patient has no history of interstitial lung disease.

[0021] In some embodiments of combined use, BT5528 or a pharmaceutical composition containing BT5528 and nivolumab are administered separately. In some embodiments, BT5528 or a pharmaceutical composition containing BT5528 and nivolumab are administered sequentially. 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.

[0022] In some embodiments, BT5528 and nivolumab are administered by separate infusions. In some embodiments, BT5528 and nivolumab are administered by sequential infusions. In some embodiments, BT5528 is administered by an initial infusion, followed by an infusion of nivolumab. In some embodiments, nivolumab is administered by an initial infusion, followed by an infusion of BT5528.

[0023] 4. General Description of Specific Embodiments of the Invention In some embodiments, the method of the present invention comprises administering a pharmaceutical composition comprising BT5528 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable additive or carrier. In some embodiments, the pharmaceutical composition comprises about 21.2 mg of BT5528 or a pharmaceutically acceptable salt thereof.

[0024] 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 freeze-dried powder. In some embodiments, the solid pharmaceutical composition of the present invention is granules.

[0025] 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 vehicle or solvent. In some embodiments, the acceptable vehicle or solvent is selected from sterile water, Ringer's solution, USP, and isotonic sodium chloride solution. In some embodiments, the acceptable vehicle or solvent is sterile water. In some embodiments, the acceptable vehicle or solvent is a sterile injectable medium. In some embodiments, the liquid pharmaceutical composition contains about 2 to 4 mg / mL of BT5528 or a pharmaceutically acceptable salt thereof.

[0026] In some embodiments, the pharmaceutically acceptable additive or carrier includes a buffer. In some embodiments, the buffer is selected from phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures 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, cellulosic substances, polyethylene glycol, sodium carboxymethylcellulose, 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.

[0027] In some embodiments, the buffer is an amount that adjusts the pH of the pharmaceutical composition of the present invention to about 6 to 8. In some embodiments, the buffer is histidine hydrochloride in an amount of about 1 to 3 mg per 1 mg of BT5528 or a pharmaceutically acceptable substance thereof. In some embodiments, the histidine hydrochloride is in an amount of about 1.31 or 2.62 mg per 1 mg of BT5528 or a pharmaceutically acceptable substance thereof. In some embodiments, the liquid pharmaceutical composition of the present invention contains histidine hydrochloride at a concentration of about 5.24 mg / mL.

[0028] In some embodiments, the liquid pharmaceutical composition of the present invention has a pH of about 6 to 8. In some embodiments, the liquid pharmaceutical composition of the present invention has a pH of 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 liquid pharmaceutical composition of the present invention has a pH of about 6.5 or 7.0.

[0029] In some embodiments, the pharmaceutically acceptable additive or carrier includes an inert pharmaceutically acceptable additive or carrier. In some embodiments, the inert pharmaceutically acceptable additive or carrier is sodium citrate or dicalcium phosphate. In some embodiments, the inert pharmaceutically acceptable additive or carrier is an excipient or filler. In some embodiments, the excipient or filler is starch, lactose, sucrose, glucose, mannitol, or silicic acid. In some embodiments, the inert pharmaceutically acceptable additive or carrier is a binder. In some embodiments, the binder is carboxymethylcellulose, alginate, gelatin, polyvinylpyrrolidinone, sucrose, or acacia. In some embodiments, the inert pharmaceutically acceptable additive or carrier is selected from sucrose, trehalose, dextrose, or a combination thereof. In some embodiments, the inert pharmaceutically acceptable additive or carrier is sucrose.

[0030] In some embodiments, the amount of an inert, pharmaceutically acceptable additive or carrier (e.g., sucrose) is about 10 to 35 mg of BT5528 or a pharmaceutically acceptable substance thereof. In some embodiments, the amount of an inert, pharmaceutically acceptable additive or carrier (e.g., sucrose) is about 15 or 30 mg of BT5528 or a pharmaceutically acceptable substance thereof. In some embodiments, the liquid pharmaceutical composition contains an inert, pharmaceutically acceptable additive or carrier (e.g., sucrose) at a concentration of about 60 mg / mL.

[0031] In some embodiments, the pharmaceutically acceptable additive or carrier includes 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 a combination thereof). In some embodiments, the surfactant is a poloxamer (e.g., poloxamer-188); TritonTM ;Sodium dodecyl 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 betaine, lauroamidopropyl betaine, cocamidopropyl betaine, linoleamidopropyl betaine, myristamidopropyl betaine, palmidopropyl betaine, isosteramidopropyl betaine (e.g., lauroamidopropyl), myristalnidopropyl-, palmidopropyl- or isosteramidopropyl-dimethylamine;Sodium methyl cocoyl-, or disodium methyl ophthalic taurate;and Monaquat TM The surfactant is selected from series (Mona Industries, Inc., Paterson, NJ), polyethyl glycol, polypropyl glycol, and copolymers of ethylene and propylene glycol (e.g., Pluronic, PF68). In some embodiments, the surfactant is polysorbate 20.

[0032] In some embodiments, the surfactant (e.g., polysorbate 20) is in an amount of about 0.01 to 0.15 mg of BT5528 or a pharmaceutically acceptable form thereof. In some embodiments, the surfactant (e.g., polysorbate 20) is in an amount of about 0.025, 0.05, or 0.1 mg of BT5528 or a pharmaceutically acceptable form thereof. In some embodiments, the liquid pharmaceutical composition contains the surfactant (e.g., polysorbate 20) at a concentration of about 0.1 or 0.2 mg / mL.

[0033] In some embodiments, the pharmaceutically acceptable additive or carrier includes an osmotic regulator. In some embodiments, the osmotic regulator is sodium chloride, dextrose, calcium chloride, or a combination thereof. In some embodiments, the osmotic regulator is dextrose. In some embodiments, the osmotic regulator is sodium chloride. In some embodiments, the osmotic regulator is a combination of sodium chloride and dextrose.

[0034] 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 is BT5528 or its pharmaceutically acceptable salts; Approximately 1.31–2.62 mg of histidine hydrochloride per 1 mg of BT5528 or a pharmaceutically acceptable equivalent; Approximately 15-30 mg of sucrose per 1 mg of BT5528 or a pharmaceutically acceptable equivalent; and Approximately 0.05–0.1 mg of polysorbate 20 per 1 mg of BT5528 or its pharmaceutically acceptable equivalent. Includes.

[0035] In some embodiments, the present invention is Approximately 21.2 mg of BT5528 or a pharmaceutically acceptable salt thereof; Approximately 55.5 mg of histidine hydrochloride per 1 mg of BT5528 or a pharmaceutically acceptable equivalent; Approximately 636 mg of sucrose per 1 mg of BT5528 or a pharmaceutically acceptable equivalent; and Approximately 1.06–2.12 mg of polysorbate 20 per 1 mg of BT5528 or a pharmaceutically acceptable equivalent. The present invention provides a solid pharmaceutical composition which is a freeze-dried powder containing the following:

[0036] In some embodiments, the present invention is Approximately 21.2 mg of BT5528 or a pharmaceutically acceptable salt thereof; Approximately 27.8 mg of histidine hydrochloride per 1 mg of BT5528 or a pharmaceutically acceptable equivalent; Approximately 318 mg of sucrose per 1 mg of BT5528 or a pharmaceutically acceptable equivalent; and Approximately 1.06 mg of polysorbate 20 per 1 mg of BT5528 or a pharmaceutically acceptable equivalent. The present invention provides a solid pharmaceutical composition which is a freeze-dried powder containing the following:

[0037] In some embodiments, the present invention is Approximately 2-4 mg / mL of 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 The present invention provides a liquid pharmaceutical composition containing the following:

[0038] 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., physiological saline or 5% dextrose). In some embodiments, the present invention provides a liquid pharmaceutical composition prepared by reconstituting the solid pharmaceutical composition of the present invention in water and subsequently diluting it with 5% dextrose. In some embodiments, the liquid pharmaceutical composition is diluted in an IV bag of 5% dextrose for IV administration. In some embodiments, the liquid pharmaceutical composition in the IV bag of 5% dextrose is stored at room temperature (about 20-25°C) for up to about 4 hours before IV administration. In some embodiments, the liquid pharmaceutical composition in the IV bag of 5% dextrose is stored under refrigerated conditions (about 2-8°C) for up to about 20 hours before IV administration. In some embodiments, the liquid pharmaceutical composition in the IV bag of 5% dextrose is stored under refrigerated conditions (about 2-8°C) for up to about 20 hours before IV administration, and then stored at room temperature (about 20-25°C) for up to 4 hours.

[0039] 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 dextrose. In some embodiments, the present invention provides a liquid pharmaceutical composition comprising BT5528 or a pharmaceutically acceptable salt thereof, histidine hydrochloride, sucrose, polysorbate 20, dextrose, and water. In some embodiments, the components of the pharmaceutical composition are as described above in terms of amounts, concentrations, and ratios.

[0040] In some embodiments, the method of the present invention includes intravenous administration of the pharmaceutical composition of this specification to a patient. In some embodiments, the pharmaceutical composition of the present invention is administered by IV injection. In some embodiments, the pharmaceutical composition of the present invention is administered by IV infusion. In some embodiments, the IV infusion of the pharmaceutical composition of the present invention lasts for about 5 to 30 minutes. In some embodiments, the IV infusion of the pharmaceutical composition of the present invention lasts for about 30 to 90 minutes. In some embodiments, the IV 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 IV infusion of the pharmaceutical composition of the present invention lasts for about 60 minutes. In some embodiments, the IV infusion of the pharmaceutical composition of the present invention lasts for about 2, 2.5, 3, 3.5, or 4 hours.

[0041] 5. Dosage regimen 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 every week. In some embodiments, the pharmaceutical composition of the present invention is administered to the patient every two weeks.

[0042] In some embodiments, the treatment cycle is four weeks, or a 28-day treatment cycle. In some embodiments, BT5528 is administered weekly in the 28-day treatment cycle, for example, on days 1, 8, 15, and 22. In some embodiments, BT5528 is administered every two weeks in the 28-day cycle, for example, on days 1 and 15. In some embodiments, BT5528 is administered as a liquid formulation or liquid unit dosage form.

[0043] In some embodiments, the treatment cycle is 3 weeks, or 21 days. In some embodiments, BT5528 is administered in weeks 1 and 2 of the 21-day treatment cycle, but not in week 3. In some embodiments, BT5528 is administered on days 1 and 8 of the 21-day treatment cycle. In some embodiments, BT5528 is administered once a week in the 21-day treatment cycle. In some embodiments, BT5528 is administered on days 1, 8, and 15 of the 21-day treatment cycle.

[0044] In some embodiments, BT5528 is approximately 1-27 mg / m² 2 It is administered in a dose (measured by the amount of API:BT5528). In some embodiments, BT5528 is approximately 2-20 mg / m². 2 It is administered in the following doses. In some embodiments, BT5528 is approximately 2-15 mg / m². 2 It is administered in the following doses. In some embodiments, BT5528 is approximately 2-10 mg / m². 2 It is administered in the following doses. In some embodiments, BT5528 is approximately 4-8 mg / m². 2 At this dosage, approximately 5-7 mg / m² is available upon request. 2 At this dosage, approximately 6.5 mg / m² can be added as desired. 2 It is administered in the following doses. In some embodiments, BT5528 is approximately 3-7 mg / m². 2 At this dosage, approximately 4-6 mg / m² is available upon request. 2 At this dosage, approximately 5 mg / m² is available upon request. 2 It is administered in the following doses. In some embodiments, BT5528 is administered at approximately 2.2, 4.4, 7.3, 11, 14.6, or 19.4 mg / m². 2 It is administered in the following doses. In some embodiments, BT5528 is administered at 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 It is administered in the following doses. In some embodiments, BT5528 is about 1-10 or 10-20 mg / m². 2It is administered in the following doses. In some embodiments, BT5528 is administered at approximately 21, 22, 23, 24, 25, 26, or 27 mg / m². 2 It is administered in the following dose.

[0045] In some embodiments, the method of the present invention involves administering BT5528 or a pharmaceutical composition containing BT5528 to a patient at a dose of approximately 4-8 mg / m² every two weeks. 2 The dosage is approximately 5-7 mg / m², depending on the desired amount. 2 The dosage is approximately 6.5 mg / m², depending on preference. 2 This includes administering it in the specified dose.

[0046] In some embodiments, the present invention involves administering BT5528 to a patient at a dose of approximately 4-8 mg / m² every two weeks. 2 The dosage is approximately 5-7 mg / m², depending on the desired amount. 2 The dosage is approximately 6.5 mg / m², depending on preference. 2 The present invention provides a method for treating solid tumors in human patients, including administering them in a dose.

[0047] In some embodiments, the method of the present invention involves administering BT5528 or a pharmaceutical composition containing BT5528 to a patient at approximately 6.5 mg / m² every two weeks. 2 This includes administering it in the specified dose.

[0048] In some embodiments, the present invention involves administering BT5528 to a patient at approximately 6.5 mg / m² every two weeks. 2 The present invention provides a method for treating solid tumors in human patients, including administering them in a dose.

[0049] In some embodiments, the present invention involves administering BT5528 or a pharmaceutical composition containing BT5528 to a patient at a dose of approximately 4-8 mg / m² every two weeks. 2 At this dosage, approximately 5-7 mg / m² is available upon request. 2 At this dosage, approximately 6.5 mg / m² can be added as desired. 2 The present invention provides a method for treating urothelial carcinoma in human patients, including administering a certain dose. In some embodiments, urothelial carcinoma is EphA2-positive, i.e., associated with EphA2 expression.

[0050] In some embodiments, the present invention administers BT5528 or a pharmaceutical composition containing BT5528 to a patient at a dose of approximately 6.5 mg / m² every two weeks. 2 The present invention provides a method for treating urothelial carcinoma in human patients, including administering a certain dose. In some embodiments, urothelial carcinoma is EphA2-positive, i.e., associated with EphA2 expression.

[0051] In some embodiments, the present invention involves administering BT5528 or a pharmaceutical composition containing BT5528 to a patient at a dose of approximately 4-8 mg / m² every two weeks. 2 At this dosage, approximately 5-7 mg / m² is available upon request. 2 At this dosage, approximately 6.5 mg / m² can be added as desired. 2 The present invention provides a method for treating ovarian cancer in human patients, comprising administering a certain dose. In some embodiments, ovarian cancer is EphA2-positive, i.e., associated with EphA2 expression.

[0052] In some embodiments, the present invention administers BT5528 or a pharmaceutical composition containing BT5528 to a patient at a dose of approximately 6.5 mg / m² every two weeks. 2 The present invention provides a method for treating ovarian cancer in human patients, comprising administering a certain dose. In some embodiments, ovarian cancer is EphA2-positive, i.e., associated with EphA2 expression.

[0053] 6. Patients with solid tumors In some embodiments, the solid tumor or progressive 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 progressive malignant tumor associated with EphA2 expression is the adenocarcinoma subtype of NSCLC (adeno-NSCLC).

[0054] In one embodiment, the solid tumor or progressive malignant tumor is lung cancer. In one embodiment, the lung cancer is NSCLC.

[0055] In one embodiment, the solid tumor or progressive malignant tumor is ovarian cancer. In one embodiment, the solid tumor or progressive malignant tumor is fallopian tube cancer.

[0056] In one embodiment, the solid tumor or progressive malignant tumor is breast cancer. In one embodiment, the breast cancer is triple-negative breast cancer.

[0057] In one embodiment, the solid tumor or progressive malignant tumor is head and neck cancer.

[0058] In one embodiment, the solid tumor or progressive malignant tumor is gastric cancer. In one embodiment, the solid tumor or progressive malignant tumor is upper gastrointestinal (GI) cancer.

[0059] In one embodiment, the solid tumor or malignant tumor is pancreatic cancer.

[0060] 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.

[0061] In one embodiment, the solid tumor or progressive malignant tumor is gastric cancer.

[0062] In one embodiment, the solid tumor or progressive malignant tumor is rectal cancer.

[0063] In one embodiment, the solid tumor or progressive malignant tumor is squamous cell carcinoma of unknown origin.

[0064] In some embodiments, solid tumors or progressive malignant tumors are associated with EphA2 expression.

[0065] In some embodiments, BT5528 is administered to patients who are at least 18 years of age. In other words, in some embodiments, the patient is an adult patient.

[0066] In some embodiments, the patient is one whose Eastern Cooperative Oncology Group (ECOG) performance status score is 0 or 1. ECOG performance status scores of 0 and 1 are described in Example 1.

[0067] In some embodiments, the patient is a patient with a disease that can be measured according to the Response Evaluation Criteria in Solid Tumors (RECIST) v1.1.

[0068] In some embodiments, the patient is a patient with acceptable organ function. In some embodiments, the patient with acceptable organ function has laboratory data selected from the following: Renal function: Creatinine clearance of ≥50 mL / min, measured by the Cockcroft-Gault formula or by 24-hour urine collection; Total bilirubin ≤ 1.5 × ULN (upper limit of normal); Serum albumin ≥ 2.5 g / dL; Aspartate aminotransferase (AST) ≤ 2.5 × ULN, or ≤ 5 × ULN if liver metastases are present; Alanine aminotransferase (ALT) ≤ 2.5 × ULN, or ≤ 5 × ULN if there are liver metastases; and International normal ratio (INR) < 1.3, or ≤ institutional ULN (anticoagulants not permitted).

[0069] In some embodiments, the patient is a patient with acceptable blood function. In some embodiments, a patient with acceptable blood function has the following laboratory data: Hemoglobin ≥ 9 g / dL; Absolute neutrophil count (ANC) ≥ 1500 cells / mm³ 3 and Platelet count ≥75,000 cells / mm 3 .

[0070] In some embodiments, the patient does not have red blood cell or platelet transfusions or growth factors within four weeks of the first administration of the pharmaceutical composition of the present invention.

[0071] In some embodiments, the patient is a woman of pregnancy who is pregnant (WOCBP) and has tested negative on a pregnancy test (negative serological test at screening and negative urine or serological test within 3 days prior to the first dose of BT5528).

[0072] In some embodiments, the patient has a metastatic, recurrent, histologically confirmed malignant solid tumor, has exhausted all appropriate treatment options according to local guidelines, has failed at at least one previous line of treatment, and has evidence of radiographic progression with the most recent line of treatment.

[0073] In some embodiments, the patient has previously received immunotherapy and received their last dose at least 28 days prior to the first dose of BT5528.

[0074] In some embodiments, the patient is eligible and has histologically confirmed metastatic or recurrent disease that is non-small cell lung cancer (NSCLC), ovarian cancer, triple-negative breast cancer (TNBC), gastric / upper gastrointestinal (GI) cancer, head and neck cancer, or urothelial carcinoma; has failed or is ineligible for all appropriate treatment options according to local guidelines; and has evidence of radiographic progression in the latest lines of treatment.

[0075] In some embodiments, the patient had urothelial carcinoma and had previously received treatment with enfortumab vedotin (EV). In some embodiments, the patient showed disease progression within 6 months of initiating EV treatment.

[0076] In some embodiments, the patient has an adenocarcinoma subtype of NSCLC (adeno-NSCLC) and has exhausted all standard treatment options, including progression during or after platinum-based chemotherapy, and / or has failed at at least one previous line of treatment with 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 a driver mutation disorder. In some embodiments, the patient has not received immunotherapy at least 28 days prior to the first dose of the pharmaceutical composition of the present invention. In some embodiments, the patient has not received chemotherapy within 14 days prior to the first dose of the pharmaceutical composition of the present invention. In some embodiments, the patient has not received anticancer treatment within 28 days or within 5 half-lives prior to the first dose of the pharmaceutical composition of the present invention, whichever is shorter. In some embodiments, the patient has prior toxicity that has resolved to Grade 1 according to the Common Terminology Criteria for Adverse Events (CTCAE) v5.0 (except for alopecia, which must be Grade 2 or less).

[0077] In some embodiments, patients have not undergone experimental treatment within four weeks of the first dose of the pharmaceutical composition of the present invention.

[0078] In some embodiments, the patient is not currently receiving treatment with potent inhibitors or inducers of CYP3A4, or potent inhibitors of P-gp, including herbs or food products.

[0079] In some embodiments, the patient is not sensitive to any of the components of the pharmaceutical composition of the present invention or to monomethyl auristatin E (MMAE).

[0080] In some embodiments, the patient does not have a serious medical condition, a life-threatening illness, an active, uncontrolled infection, or organ system dysfunction (e.g., ascites, coagulation disorders, encephalopathy).

[0081] In some embodiments, the patient has not undergone any major surgery (excluding the placement of vascular access) within four weeks of the first administration of the pharmaceutical composition of the present invention.

[0082] In some embodiments, the patient has not received a live vaccine within 30 days of the first dose of the pharmaceutical composition of the present invention.

[0083] In some embodiments, the patient does not have uncontrolled symptomatic brain metastases. In some embodiments, the patient has a stable neurological condition after at least 4 weeks of topical treatment without steroids, or with prednisone or equivalent at a daily dose of less than 10 mg or an equivalent stable or tapering dose.

[0084] In some embodiments, the patient does not have untreated CNS metastases. In some embodiments, the patient has treated CNS metastases and a) the CNS metastases are clinically stable for at least 6 weeks prior to screening; b) if steroid treatment is required for CNS metastases, the subject has received a stable or tapering dose of prednisone or equivalent ≤20 mg / day for at least 2 weeks; c) baseline scans show no evidence of new or expanding brain metastases; d) the subject does not have leptomeningeal disease; and e) the subject does not have neurological dysfunction that would interfere with the assessment of neurological and other AEs.

[0085] In some embodiments, the patient does not have uncontrolled hypertension (systolic blood pressure [BP] ≥ 139 mmHg; diastolic BP ≥ 89 mmHg) prior to the first administration of the pharmaceutical composition of the present invention. In some embodiments, the patient has had stable, controlled hypertension for at least three months prior to the first administration of the pharmaceutical composition of the present invention.

[0086] In some embodiments, the patient does not have uncontrolled hypertension (not responding to intervention, systolic blood pressure [BP] ≥ 160 mmHg; diastolic BP ≥ 100 mmHg) at the time of screening or prior to the first administration of the pharmaceutical composition of the present invention. In some embodiments, the patient has had stable, controlled hypertension for at least three months prior to the first administration of the pharmaceutical composition of the present invention.

[0087] 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 recorded within six months prior to the first administration of the pharmaceutical composition of the present invention.

[0088] In some embodiments, the patient does not have a mean resting-adjusted QT interval (QTcF) > 470 milliseconds within 6 months prior to the first administration of the pharmaceutical composition of the present invention.

[0089] In some embodiments, the patient does not have any factors that increase the risk of QTc prolongation or arrhythmic events, such as heart failure, hypokalemia, congenital long QT syndrome, a family history of long QT syndrome, or sudden death of unknown cause before the age of 40, or any concomitant medications known to prolong the QT interval, within 6 months prior to the first administration of the pharmaceutical composition of the present invention.

[0090] In some embodiments, the patient does not have any clinically significant abnormalities in resting electrocardiogram (ECG) rhythm, conduction, or morphology, such as complete left bundle branch block or third-degree cardiac block, within six months prior to the first administration of the pharmaceutical composition of the present invention.

[0091] In some embodiments, the patient does not have human immunodeficiency virus (HIV) or acquired immunodeficiency syndrome (AIDS).

[0092] In some embodiments, the patient has well-controlled HIV and meets all of the following criteria at the time of inclusion: a.CD4+ count ≥350 cells / μL; b. HIV viral load < 400 copies / mL; c. No history of opportunistic infections within the last 12 months; d. The patient has been receiving established ART for at least four weeks. The use of ART should be considered on a case-by-case basis in conjunction with medical monitoring.

[0093] In some embodiments, the patient does not have a positive hepatitis B surface antigen and / or anti-hepatitis B core antibody.

[0094] In some embodiments, the patient has a negative polymerase chain reaction (PCR) assay and is receiving appropriate antiviral treatment.

[0095] In some embodiments, if a patient tests positive for hepatitis C virus (HCV) antibodies, the patient has an active hepatitis C infection with a positive viral load.

[0096] In some embodiments, the patient has been treated for hepatitis C infection and has a persistent virological response of ≥12 weeks.

[0097] In some embodiments, the patient has not had a thromboembolic event and / or bleeding disorder (e.g., deep vein thrombosis [DVT] or pulmonary embolism [PE]) within three months prior to the first administration of the pharmaceutical composition of the present invention.

[0098] In some embodiments, the patient has not had another malignant tumor within three years prior to the first administration of the pharmaceutical composition of the present invention. In some embodiments, the patient has no residual disease from a previously diagnosed malignant tumor (excluding basal cell carcinoma, squamous cell carcinoma of the skin, cervical intraepithelial neoplasia / cervical carcinoma in situ, or melanoma or ductal carcinoma in situ that has been appropriately treated for curative purposes).

[0099] In some embodiments, the patient has not had systemic anti-infective treatment or fever within the last 14 days prior to the first administration of the pharmaceutical composition of the present invention. [Examples]

[0100] Example 1. Phase I / II study of the safety, pharmacokinetics, and preliminary clinical activity of BT5528 in patients with advanced malignant tumors associated with EphA2 expression.

[0101] 2.1. Purpose Main purpose The primary objectives of the gradual increase (Parts A-1 and A-2) are as follows: • Evaluate the safety and tolerability of BT5528 administered weekly in patients with advanced solid tumors associated with EphA2 expression and / or specific tumors identified as positive for EphA2 tumor expression, both as monotherapy (Part A-1) and in combination with nivolumab (Part A-2). Define the MTD of BT5528 as monotherapy (Part A-1) and in combination with nivolumab (Part A-2), and determine one or more recommended Phase II doses if observed.

[0102] The main objectives of the expanded trial (Part B-1) are as follows: To evaluate the clinical activity of BT5528 as monotherapy in patients with solid tumors historically known to have high EphA2 expression in one or more R2PD 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).

[0103] Secondary purpose The secondary objectives of the progressive format of this examination (Parts A-1 and A-2) are as follows:

[0104] The secondary objectives of the expanded (Part B) trial are to evaluate preliminary signals of antitumor activity achieved with BT5528 administration in patients with advanced solid tumors associated with EphA2 expression and / or specific tumors identified as positive for EphA2 tumor expression, both as monotherapy (Part A-1) and in combination with nivolumab (Part A-2): To evaluate the safety and tolerability of BT5528 as monotherapy in patients with tumors historically known to have high EphA2 expression, and to assess whether tumor EphA2 expression levels predict clinical activity of BT5528 monotherapy in patients with tumors historically known to have high EphA2 expression.

[0105] The secondary objectives of all parts of the exam are as follows: • Determine the pharmacokinetic (PK) parameters of BT5528 and MMAE (if necessary). • To determine the incidence of anti-drug antibodies (ADAs).

[0106] 2.2. Examination Design This is a Phase I / II human, open-label, dose-escalation study of BT5528 administered as monotherapy (Parts A-1 and B) and in combination with nivolumab (Part A-2). The study consists of two parts: Part A, dose escalation, and Part B, dose expansion.

[0107] 2.3. Investigational drug, dosage, and mode of administration: In Part A, BT5528 was administered intravenously as an escalating dose over 1 hour, and in Part B, it was administered according to RP2D. Nivolumab was administered intravenously at a dose of 480 mg every 4 weeks or 240 mg every 2 weeks in Part A.

[0108] 2.4. Inclusion Criteria - All Patients: Patients must meet the following criteria to be included in the study: 1. Prior to performing any procedures, sampling, or analysis specific to the study, a written informed consent signed and dated by the patient or legal guardian, in accordance with local guidelines, has been obtained. If a patient refuses to participate in any component of the trial (e.g., tumor biopsy), the patient will not incur any penalty or loss of benefit, nor will they be excluded from other aspects of the trial. 2. At least 18 years of age at the time of signing the informed consent form. 3. The US East Coast Cancer Clinical Trials Group (ECOG) Performance Status Score is 0 or 1 [Table 1] 4. The patient must have a disease that can be measured according to the Criteria for Evaluation of Effectiveness of Solid Tumors (RECIST) v1.1. 5. Acceptable organ function as demonstrated by the following test data: • Renal function is as follows: Creatinine clearance of ≥50 mL / min, measured by the Cockcroft-Gault formula or by 24-hour urine collection. • Total bilirubin ≤ 1.5 × ULN (upper limit of normal range) • Serum albumin ≥ 2.5 g / dL • Aspartate aminotransferase (AST) ≤ 2.5 × ULN, or ≤ 5 × ULN if liver metastases are present. • Alanine aminotransferase (ALT) ≤ 2.5 × ULN, or ≤ 5 × ULN if liver metastases are present. • International Normal Ratio (INR) < 1.3, or ≤ Facility ULN 6. Acceptable blood function (no red blood cell or platelet transfusion or growth factor required within 4 weeks of the first dose of BT5528): • Hemoglobin ≥ 9 g / dL • Absolute neutrophil count (ANC) ≥ 1500 cells / mm³ 3 ·Platelet count ≧75,000 cells / mm 3 7. Negative pregnancy test for women of childbearing potential (WOCBP) (negative serological test at screening and negative urine or serological test within 3 days prior to the first dose of T5528). Male patients with a female partner of childbearing potential and female patients of childbearing potential are required to use highly effective contraception (oral and hormonal contraceptives are permitted) with a failure rate of less than 1% during participation in the study, for 6 months after the last dose of BT5528 (BT5528 IB) or for 5 months after the last dose of nivolumab (nivolumab USPI or SmPC) for patients continuing treatment with nivolumab alone after discontinuing treatment with BT5528. Male patients must also refrain from sperm donation during participation in the study and for 5 or 6 months after the last dose of either nivolumab or BT5528, respectively, and women must not breastfeed or donate eggs during that time. 8. All patients must have tumor tissue (fresh or preserved) available for analysis of EphA2 tumor expression and other biomarkers. If tumor tissue is not available, 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. 9. Mean life expectancy ≥ 12 weeks prior to initiation of BT5528 treatment, as determined by the investigator. 10. You must have the willingness and ability to comply with protocols and test procedures.

[0109] Further inclusion criteria - Part A only 11. The patient must have a metastatic, recurrent, histologically confirmed malignant solid tumor, have exhausted all appropriate treatment options according to local guidelines, have failed at least one previous line of treatment, and have evidence of radiographic progression on the most recent line of treatment. If applicable, for patients who have previously received immunotherapy, the last dose must have been received at least 28 days prior to the first dose of the BT5528 study treatment. Pancreatic cancer patients are acceptable, but are planned to represent less than 50% of all patients enrolled in each escalation cohort. The sponsor or SRC may decide to require enrollment of a specific tumor type or subtype at any point during the escalation if they believe it is necessary to enhance the evaluation of biomarkers, safety, or pharmacokinetics in a particular tumor type. 12. Patients enrolled in cohort A1-8 or later for Part A-1 monotherapy, or in cohort A2-3 or later for Part A-2 combination therapy, must have tumor tissue (fresh or preserved) available in the central laboratory for prospective or retrospective analysis of EphA2 tumor expression, but may be enrolled without prior confirmation of EphA2 expression.

[0110] Further inclusion criteria - Part B monotherapy cohort 13. Patients with metastatic or recurrent disease histologically confirmed to be non-small cell lung cancer (NSCLC), ovarian cancer, triple-negative breast cancer (TNBC), gastric / upper gastrointestinal (GI) cancer, head and neck cancer, or urothelial carcinoma are eligible; they must have failed or been ineligible for all appropriate treatment options according to local guidelines; and they must have evidence of radiographic progression on the most recent line of treatment. 14. Patients with urothelial carcinoma who have previously received treatment with enfortumab vedotin (EV) are eligible for the trial. Patients who received EV and experienced disease progression within 6 months of treatment initiation will be planned for less than 50% of all patients enrolled in the cohort.

[0111] 2.5. Exclusion criteria - all patients: Patients who meet any of the following criteria will be excluded from the study: 1. Chemotherapy must be administered within 14 days prior to the first dose of the study treatment. For other anticancer treatments, the treatment must be administered within 28 days or within 5 half-lives, whichever is shorter. For immunotherapy including immune checkpoint inhibitors, the treatment must be administered within 28 days prior to the first dose of the study treatment. 2. Experimental treatment within 4 weeks of the first dose of the BT5528 test treatment. 3. Previous toxicity must be resolved to Grade 1 according to the Common Terminology Criteria for Adverse Events (CTCAE) v5.0 (excluding alopecia, which may be Grade 2). 4. Current treatment with potent inhibitors or inducers of CYP3A4, or potent inhibitors of P-gp, including herbs and food products. 5. Known susceptibility to either the test drug or any component of monomethyl auristatin E (MMAE). 6. A serious medical condition, life-threatening illness, active, uncontrolled infection or organ dysfunction (e.g., ascites, coagulation disorder, encephalopathy), or any other reason that, in the investigator's opinion, could impair patient safety or interfere with or impair the integrity of the study results, including consideration of gastrointestinal, skin, and pulmonary comorbidities, and review of screening chest CT to confirm the absence of clinically significant comorbidities. 7. Patients must undergo major surgery (excluding placement of vascular access) within four weeks of the first dose of the BT5528 study treatment and must be fully recovered before the start of the study treatment. 8. Administer live vaccine within 30 days of the test procedure. 9. Untreated CNS metastases. Subjects with treated CNS metastases are permitted to be tested if all of the following are true: a. CNS metastases are clinically stable for at least 6 weeks prior to screening. b. If steroid treatment is required for CNS metastases, the subject should receive a stable or tapered dose of prednisone or equivalent of ≤20 mg / day for at least two weeks. c. Baseline scan does not show evidence of new or expanded brain metastases. d. The subject does not have piatric disease. e. The subject must not have neurological dysfunction that would interfere with the assessment of neurological and other adverse events. 10. Uncontrolled hypertension at screening or prior to initiation of investigational drug (systolic BP ≥ 160 mmHg or diastolic BP ≥ 100 mmHg that does not respond to intervention). 11. Any medical history or current evidence of any condition, treatment or laboratory abnormality that could interfere with the trial results, prevent patient participation, or, in the opinion of the investigator, be in the patient's best interest in participating, including but not limited to: a. Patients with a 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 recorded within 6 months prior to the first dose of BT5528, or; i. Mean resting-adjusted QT interval (QTcF) > 470 milliseconds ii. Any factors that increase the risk of QTc prolongation or arrhythmic events, such as heart failure, hypokalemia, congenital long QT syndrome, family history of long QT syndrome, or sudden death of unknown cause before age 40, or any concomitant medications known to prolong the QT interval. iii. Any clinically significant abnormalities in the rhythm, conduction, or morphology of the resting electrocardiogram (ECG) (as assessed by the investigator), e.g., complete left bundle branch block, third-degree cardiac block. 12. Known human immunodeficiency virus (HIV) or acquired immunodeficiency syndrome (AIDS) Note: Well-controlled HIV is permitted if the patient meets all of the following criteria: a) CD4+ cell (CD4+) count ≥350 cells / μL; b) HIV viral load < 400 copies / mL; c) No history of opportunistic infection within the last 12 months; d) The patient has received established antiretroviral therapy (ART) for at least four weeks. The use of antiretroviral therapy is permitted, but should be considered on a case-by-case basis in conjunction with medical monitoring. 13. Patients who have positive hepatitis B surface antigen and / or anti-hepatitis B core antibody. Patients with negative polymerase chain reaction (PCR) assay are permitted to receive appropriate antiviral treatment. 14. Active hepatitis C infection with a positive viral load, provided that the hepatitis C virus (HCV) antibody test is positive (if the antibody test is negative, the viral load is not applicable). Patients treated for hepatitis C infection may be included if they have recorded a persistent virological response of ≥12 weeks. 15. Thromboembolic events and / or bleeding disorders (e.g., deep vein thrombosis [DVT] or pulmonary embolism [PE]) within 3 months prior to the first dose of the BT5528 study treatment. 16. A history of pneumonia with residual symptoms. 17. A history of another malignant tumor within three years prior to the first dose of BT5528, or any evidence of residual disease from a previously diagnosed malignant tumor (excluding basal cell carcinoma, squamous cell carcinoma of the skin, cervical intraepithelial neoplasia / cervical carcinoma in situ, or melanoma in epidermis or ductal carcinoma in situ that has been appropriately treated for curative purposes). 18. Systemic anti-infective treatment or fever within the last 14 days prior to the first dose of BT5528 test treatment. 19. Psychological, familial, social, or geographical conditions that do not permit compliance with the procedure and / or follow-up procedures outlined therein.

[0112] Further exclusion criteria Part A-2: Nivolumab combination cohort 20. Previous intolerance to immune checkpoint inhibitors 21. Known hypersensitivity to checkpoint inhibitor therapy 22. Previous organ transplants (including allogeneic and allogeneic transplants) 23. Diagnosis of clinically relevant immunodeficiency 24. Active systemic infections requiring treatment 25. Prednisone equivalent or other potent immunosuppressants exceeding 10 mg per day 26. History of autoimmune diseases other than alopecia or vitiligo.

[0113] 2.6. Correlation Test: All patients are required to provide preserved tumor material or fresh tumor biopsy for evaluation of EphA2 expression levels and further molecular genetic characterization (i.e., evaluation of specific somatic mutations). This material should be provided as a tissue block or 10–15 paraffin-immersed, unstained slides.

[0114] Enrollment in cohorts A2-3 or later for Part A-1 monotherapy or Part A-2 combination therapy requires the presence of tumor tissue (fresh or preserved) to be submitted to the central laboratory for prospective or retrospective analysis of EphA2 tumor expression, although enrollment may be done without prior confirmation of EphA2 expression. The SRC may review the EphA2 expression requirement before initiating treatment if the individual tumor type or subtype has already shown signs of efficacy.

[0115] For Part B (all expanded cohorts), EphA2 expression analysis will be performed retrospectively at the beginning, and confirmation of EphA2 expression before the study treatment is not required. However, if efficacy analyses performed for individual indications or cohorts indicate the need to accumulate patients with EphA2 expression in a particular tumor indication cohort, confirmation of EphA2 expression before initiating the study treatment will be required for those cohorts.

[0116] To investigate the intratumoral PK / pharmacodynamic effects of BT5528, tumor biopsies will be taken post-administration, subject to patient consent. Post-administration tumor biopsies are optional for all patients. Post-administration tumor biopsies may be taken between 4 hours after the end of infusion (EOI) and 7 days after any BT5528 infusion within Cycle 1, or, if not feasible, any subsequent cycle is acceptable. See the Evaluation Schedule (SoA) for further details.

[0117] Blood samples will also be collected before and after administration to evaluate pharmacodynamic, response, and treatment resistance biomarkers, such as somatic mutations in circulating tumor DNA (ctDNA), ADA, and pharmacological genomic analysis.

[0118] 2.7.Statistical methods: Dose escalation (applies separately to A-1 and A-2): The actual number of dose levels considered in this study depends on the determination of the unacceptable dose based on dose-limiting toxicity (DLT). MTD is defined based on DLT (see Section 5). Using other safety data, as well as any trends in PK profile and antitumor activity observed during the conduct of the study, the RP2D is determined to be less than or equal to the MTD (or, if no MTD is designated, less than or equal to the maximum dose administered [MAD], which is defined as the highest dose tested according to the escalation).

[0119] A 3+3 design will be used for the first two dose levels. At least three evaluable patients will be enrolled at each dose level and evaluated for 28 days before any dose escalation to the next dose level may occur. After confirming tolerability at dose level 1, a dose escalation of less than 100% to dose level 2 will be permitted. Treatment cycles will be performed sequentially according to the State of Assessment (SoA). If one patient experiences a dose limit (DLT), three additional patients will be treated with the same dose. Evaluation of the cohort of at least three patients who have completed one treatment cycle (28 days) is required before proceeding to the next dose level. Further details are provided in Section 5.

[0120] Following evidence of tolerability at the first two dose levels, all subsequent dose escalations are based on a type of continuous reassessment (CRM) using a two-parameter Bayesian logistic regression model (BLRM) and the principle of escalation with overdose control (EWOC). The EWOC principle dictates that the next highest escalation dose level includes the dose with the highest posterior probability of causing a DLT among doses that meet the overdose criteria, such as having an ≤25% probability of being deemed unsafe (DLT rate ≥33%) at the target intervals (20%, 33%). The BLRM is applied to cumulative DLT / safety data, and the results are provided to the SRC, which reviews this data and makes recommendations for more precise dose escalation.

[0121] Each cohort in each RP2D of Part B uses a Simon two-stage design with p0=0.11 and p1=0.30, a one-sided alpha of 0.05, and 80% power, where p0 and p1 are the null and alternative hypotheses for overall response rate (ORR). If three or more patients out of the first 14 patients recruited show an OR, the cohort is considered by the sponsor to determine whether it is possible to proceed to stage two to enroll an additional 26 patients; otherwise, the cohort is terminated. The additional 26 patients will be enrolled in only up to two Part B cohorts.

[0122] The efficacy results from Part A of the trial may be used to identify specific cohort groups based on the EphA2 status. For all cases in Part B of the trial, efficacy will be determined based on operations ratios (OR).

[0123] As a result of these changes, the maximum number of patients recruited for the trial is 288, with 72 patients from Part A-1, 24 from Part A-2, and up to 192 patients in the Part B cohort, provided that the clinical efficacy criteria based on Simon's Stage 2 analysis of the above patients are met for all indications tested.

[0124] For ORR and CBR, estimates and associated 95% confidence intervals (CIs) (based on the Clopper-Pearson method) are calculated. For PFS and OS, Kaplan-Meier curves are constructed, providing the median time to event and associated 95% confidence intervals.

[0125] Example 3. Top-line results at the end of Phase I gradual increase BT5528 exhibits advanced antitumor activity in advanced solid tumors with a promising overall safety / tolerability profile. The antitumor activity of BT5528 was tested in cancer patients who had been treated extensively previously. A 67% ORR was observed in urothelial carcinoma patients with tumors associated with EphA2 expression (Figure 2). A novel relationship exists between antitumor activity and EphA2 expression (Figure 3A). BT5528 was also shown to reduce tumor volume in female patients with urothelial carcinoma who had received four previous lines of treatment (Figure 4). The novel safety profile of BT5528 distinguishes it from other EphA2 target molecules. • No clinical bleeding / coagulation disorders related to IMP. • No evidence of IMP-related ocular toxicity or clinically significant skin toxicity. • No reports of ILD / pneumonia in Part B. • No significant hepatotoxicity. • GI and hematological toxicity are dose-dependent and manageable. - Improvement in the rate / severity of CINV through antiemetic prophylaxis - Use of rare growth factors • BP / Vital Signs: No concerning trends • ECG: There is no tendency for concerns about the QT issue. [Brief explanation of the drawing]

[0126] [Figure 1-6] (Not mentioned in the original text)

Claims

1. A method comprising administering to a human patient BT5528 or a pharmaceutical composition containing BT5528 for treatment of the human patient.

2. A method comprising administering BT5528 or a pharmaceutical composition containing BT5528 for the treatment of solid tumors in human patients.

3. The method according to claim 2, wherein the solid tumor is a progressive malignant tumor.

4. The method according to claim 2 or 3, wherein a solid tumor or progressive malignant tumor is associated with EphA2 expression in a human patient.

5. BT5528 or a pharmaceutical composition containing BT5528 is administered to the patient at a dose of approximately 4-8 mg / m² every two weeks. 2 The dosage, depending on your preference, is approximately 5-7 mg / m². 2 The dosage is approximately 6.5 mg / m², depending on preference. 2 The method according to any one of claims 1 to 4, comprising administering in the specified dose.

6. The method according to any one of claims 2 to 5, wherein the solid tumor or progressive 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.

7. The method according to any one of claims 1 to 6, further comprising administering an anti-PD-1 antibody or an anti-PD-L1 antibody to a patient, wherein the anti-PD-1 antibody is optionally nivolumab.

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