Administration of a Beta-Catenin Antagonist and Methods of Use

Description

1 / 45 “ADMINISTRATION OF A BETA-CATENIN ANTAGONIST AND METHODS OF USE” FUNDAMENTALS

[0001] The Wnt / β-catenin signaling pathway regulates important cellular functions, including proliferation, differentiation, migration, and stem cell renewal, and plays essential roles in embryonic development and adult tissue homeostasis (Liu 2022). Dysregulation of this pathway often involves the constitutive activation of β-catenin, an oncogenic transcription factor implicated in the pathogenesis of many malignancies; it is associated with poor prognosis in many types of cancer (Shang 2017). Wnt / β-catenin signaling is also associated with the promotion of an immunosuppressive tumor microenvironment characterized by decreased chemokine expression, impaired dendritic cell recruitment, and a subsequent reduction in tumor-infiltrating effector T cells (Ruiz de Galarreta 2019).

[0002] In normal human cells, cytoplasmic and nuclear expression of β-catenin is tightly regulated by the adenomatous polyposis coli (APC) complex, a protein complex that targets excess β-catenin for proteolytic degradation. Loss-of-function mutations of APC complex components or gain-of-function mutations of β-catenin allow β-catenin to escape degradation and accumulate within the nucleus. Nuclear β-catenin engages the transcription factors lymphoid enhancing factor / T-cell factor (LEF / TCF) to induce the expression of oncogenes that promote cell survival, proliferation, and migration (Bugter 2021).

[0003] In cancer cells, the accumulation and retention of β-catenin within the nucleus are facilitated by the B-cell CLL / lymphoma 9 (BCL9) protein (de la Roche 2008), a coactivator that transports β-catenin to the nucleus and acts as a scaffold to recruit the enhancer protein Pygopus to the β-catenin transcription complex to enhance transcriptional activity (Mieszcanek 2019). BCL9 is overexpressed in many tumors and is Petition 870250092218, dated 09 / 10 / 2025, pp. 134 / 179 2 / 45 correlated with poor prognosis (Moor 2015, Vafaizadeh 2021, Wang 2019). Studies have shown that BCL9 / BCL9L deletion inhibits tumor growth and proliferation, having only a minimal impact on normal tissue homeostasis, suggesting that β-catenin antagonists may result in a significant antitumor effect, although they are unlikely to cause toxicity outside the tissue (Gay 2019).

[0004] Based on its critical role in promoting malignant phenotypes and its inverse correlation with disease prognosis in several types of cancer, β-catenin has been recognized as a potential therapeutic target for cancer therapy (Liu 2022). However, despite extensive ongoing research, β-catenin is conventionally considered “untreatable” and there are currently no drugs available to target Wnt / β-catenin signaling.

[0005] Previous approaches to targeting the Wnt / β-catenin pathway for cancer therapy act upstream of β-catenin regulation or at the site of β-catenin interaction with LEF / TCF, causing non-specific disruption of the canonical Wnt pathway and resulting in target toxicities in bone (Zhong 2016) and intestine (Dotan 2020). An approach that specifically disrupts the interaction between β-catenin and BCL9 is needed to avoid adverse effects such as target toxicities. SUMMARY OF THE INVENTION

[0006] Some of the main aspects of the present invention are summarized below. Additional aspects are described in the Detailed Description of the Invention, Examples, Drawings, and Claims sections of this disclosure. The description in each section of this disclosure should be read in conjunction with the other sections. Furthermore, the various embodiments described in each section of this disclosure can be combined in many different ways, and all such combinations must be within the scope of the present invention.

[0007] The disclosure provides a method of treating a solid tumor in a patient, the method comprising administering parenterally to Petition 870250092218, dated 09 / 10 / 2025, pp. 135 / 179 3 / 45 patient a pharmaceutical composition comprising an effective amount of a β-catenin peptide antagonist. A pharmaceutical composition for parenteral administration comprising an effective amount of a β-catenin peptide antagonist is also provided for use in the treatment of a solid tumor in a patient.

[0008] In one embodiment, the solid tumor is selected from the group consisting of breast cancer, colorectal cancer, cholangiocarcinoma, hepatocellular carcinoma, melanoma, non-small cell lung cancer, ovarian cancer, endometrial cancer, pancreatic adenocarcinoma, and synovial sarcoma. In certain embodiments, the patient has received prior treatment selected from the group consisting of chemotherapy, hormone therapy, radiation, targeted therapy, immunotherapy, and combinations thereof. In a specific embodiment, the peptide antagonist is administered as a neoadjuvant as a single agent or in combination. In one embodiment, the peptide antagonist is administered in combination with (i) bevacizumab and / or (ii) folinic acid, fluorouracil, and irinotecan (FOLFIRI). In one embodiment, the peptide antagonist is administered in combination with fruquintinib.

[0009] In some embodiments, the peptide antagonist comprises the amino acid sequence D FRWLLRQLARLAQLAKLTPVKLTPV (SEQ ID NO: 1). In some embodiments, the peptide antagonist is administered to the patient at a dose of approximately 0.25-12 mg / kg.

[0010] One aspect of the invention is a method of administering a β-catenin peptide antagonist to a subject, wherein the method comprises administering parenterally to the subject a pharmaceutical composition comprising the peptide antagonist, wherein the peptide antagonist comprises the sequence of D-amino acids. FRWLLRQLARLAQLAKLTPVKLTPV (SEQ ID NO: 1), and in which the peptide antagonist is administered at a dose of approximately 0.25-12 mg / kg. A pharmaceutical composition comprising a β antagonist is also provided. Petition 870250092218, dated 09 / 10 / 2025, pp. 136 / 179 4 / 45 catenin for use in a method for administering parenteral peptide antagonist to a subject, wherein the peptide antagonist comprises the D-amino acid sequence FRWLLRQLARLAQLAKLTPVKLTPV (SEQ ID NO: 1), and wherein the peptide antagonist is administered at a dose of about 0.25-12 mg / kg.

[0011] In certain methods and compositions of the invention, the peptide antagonist is administered at a dose selected from about 0.5 mg / kg, about 1 mg / kg, about 2 mg / kg, about 4 mg / kg, about 8 mg / kg and about 12 mg / kg.

[0012] In some aspects of the invention, the peptide antagonist disrupts the binding of β-catenin to the B-cell CLL / lymphoma 9 (BCL9) protein. In certain embodiments, the peptide antagonist comprises an N-terminal octanoyl group. In one particular embodiment, the peptide antagonist is ST316.

[0013] In one particular embodiment, the pharmaceutical composition is administered intravenously, for example, by means of an infusion. In embodiments where the composition is administered by infusion, the duration of the infusion may be, for example, about 30-180 minutes or about 60-90 minutes.

[0014] In some embodiments, the pharmaceutical composition is administered once a week or once every two weeks. In one embodiment, the pharmaceutical composition is administered for at least four weeks. BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 shows that ST316 binds directly to β-catenin Arm1 with greater affinity than the B-cell CLL / lymphoma 9 coactivator of transcription protein (BCL9) in a fluorescence polarization assay. Results are represented as normalized milliP (mP) versus β-catenin concentration; Figure 2 shows that ST316 inhibits the protein-protein interaction between β-catenin and BCL9 in a homogeneous proximity assay. Petition 870250092218, dated 09 / 10 / 2025, pp. 137 / 179 5 / 45 luminescent amplified (ALPHA). ST316 (circles) and BCL9 HD2 peptide (squares) inhibited the interaction of β-catenin and BCL9 with an IC50 of 1 μM; Figures 3A-3B show the attenuation of the nuclear localization of β-catenin by 3 μM or 5 μM of ST316 in HCT116 colorectal cancer cells. YY1 and GAPDH were used as markers for the nuclear and cytoplasmic fractions, respectively (Figure 3A); Figures 4A-4D show that ST316 increases the proteasomal degradation of BCL9 in Colo320 colorectal cancer cells; Figures 5A-5B show that ST316 inhibits the migration of HCT116 colorectal cancer cells through a semipermeable membrane in a 24-hour invasion assay, compared to a control peptide. Figure 5A shows cells after treatment with 1 μM of a negative control peptide (top panel) or ST316 (bottom panel). Figure 5B shows the cell count after treatment; Figures 6A-6C show that ST316 reduces spontaneous adenoma formation in a murine model of colorectal cancer; Figure 7 shows that administration of ST316 (βCAP) inhibits tumor growth in a subcutaneous xenograft tumor model using HCT116 cells. N = 6 mice per group; Figures 8A-8B show that administration of ST316 inhibits tumor growth (Figure 8A) and reduces Axin2 expression (Figure 8B) in a subcutaneous xenograft tumor model using triple-negative breast cancer (TNBC) 4T1-luc cells. N = 6 mice per group; Figures 9A-9B show the plasma concentrations of ST316 over time for Cohorts 1-4 (0.5, 1, 2 or 4 mg / ml); Figures 10A-10C show the ST316-induced reduction in the population of immunosuppressive polymorphonuclear myeloid-derived (PMN) suppressor cells (MDSCs) in cancer patients. Peripheral blood samples were collected from a patient with pancreatic ductal adenocarcinoma (PDAC) (Figure 10A) or colorectal cancer (CRC) (Figure 10B) prior to Petition 870250092218, dated 09 / 10 / 2025, pp. 138 / 179 6 / 45 (upper panels) and after treatment with 4 mg / kg ST316, QW for 3 weeks (lower panels). Blood was processed to collect lymphocytes and myeloid cells and analyzed by flow cytometry. PMN cells (CD3-, CD19-, CD14-, HLADRlow, CD11b+, and CD15+) were quantified as a percentage of myeloid cells (CD3-, CD19-) (Figure 10C). DETAILED DESCRIPTION OF THE INVENTION

[0015] The practice of the present invention may employ, unless otherwise indicated, conventional techniques of pharmaceuticals, formulation science, protein chemistry, cell biology, cell culture, molecular biology, microbiology, recombinant DNA, immunology, clinical pharmacology and clinical practice, which are within the skill of the art.

[0016] In order that the present invention may be more readily understood, certain terms are defined first. Additional definitions are set forth throughout this disclosure. Unless otherwise defined, all technical and scientific terms used in this document have the same meaning as commonly understood by one skilled in the art to which this invention pertains.

[0017] The headings provided in this document are not limitations on the various aspects of the disclosure, which can be obtained by reference to the descriptive report as a whole. Consequently, the terms defined immediately below are more fully defined by reference to the descriptive report in its entirety.

[0018] All references cited herein are incorporated herein by reference in their entirety. In addition, any instructions or catalogs from manufacturers of any products cited or mentioned herein are incorporated by reference. Documents incorporated by reference in this text, or any teachings contained therein, may be used in the practice of the present invention. Documents incorporated by reference in this text are not admitted as prior art. I. Definitions Petition 870250092218, dated 09 / 10 / 2025, pp. 139 / 179 7 / 45

[0019] It should be understood that the phraseology or terminology in this disclosure is for descriptive purposes only and not for limitation, so that the terminology or phraseology of this descriptive report should be interpreted by a person skilled in the art in the light of the teachings and guidance.

[0020] As used in this descriptive report and the accompanying claims, the singular forms “a”, “an”, “the” and “the” include plural referents, unless the context clearly indicates otherwise. The terms “a” (or “an”), as well as the terms “one or more” and “at least one” can be used interchangeably.

[0021] Additionally, “and / or” should be considered as specific disclosure of each of the two specified features or components with or without the other. Thus, the term “and / or”, as used in an expression such as “A and / or B”, is intended to include “A and B”, “A or B”, A (only) and B (only). Similarly, the term “and / or” as used in an expression such as “A, B and / or C” is intended to include A, B and C; A, B or C; A or B; A or C; B or C; A and C; B and C; A (only); B (only); and C (only).

[0022] Wherever modalities are described using the language “comprising”, analogous modalities described in terms of “consisting of” and / or “consisting essentially of” are included.

[0023] Units, prefixes, and symbols are denoted in their accepted form in the International System of Units (SI). Numerical intervals include the numbers that define the interval, and any individual value given here can serve as an endpoint for an interval that includes other individual values ​​given here. For example, a set of values ​​such as 1, 2, 3, 8, 9, and 10 is also a disclosure of a number interval from 1-10, from 1-8, from 3-9, and so on. Similarly, a disclosed interval is a disclosure of each individual (i.e., intermediate) value covered by the interval, including integers and fractions. For example, a stated interval of 5-10 is also a disclosure of 5, 6, 7, 8, 9, and 10 individually, and of 5.2, 7.5, 8.7, and so on. Petition 870250092218, dated 09 / 10 / 2025, pp. 140 / 179 8 / 45

[0024] Unless otherwise indicated, the terms “at least” and “about” preceding a series of elements should be understood as referring to each element of the series. The term “about” depends on the context in which it is used. When preceding a numerical value, it generally includes ± 10% of the stated value. For example, in one context, a concentration of about 1 mg / ml includes 0.9 mg / ml to 1.1 mg / ml. Similarly, a concentration range of about 1% to 10% (w / v) may include 0.9% (w / v) to 11% (w / v).

[0025] The terms “polypeptide,” “peptide,” and “protein” are used interchangeably in this document to refer to polymers of amino acids of any length and their salts. The polymer may be linear or branched, may comprise modified amino acids, and may be interrupted by non-amino acids. Except where otherwise indicated, for example, for the abbreviations for unusual or unnatural amino acids presented herein, the three-letter and one-letter abbreviations, as used in the art, are used herein to represent amino acid residues. Except when preceded by a “D” or in lowercase letters, the amino acid is an L-amino acid. Groups or sequences of amino acid abbreviations are used to represent peptides. Except where specifically indicated, peptides are indicated with the N-terminal on the left and the sequence is written from the N-terminal to the C-terminal.

[0026] A “retro inverse” peptide has an inverted amino acid sequence, relative to a reference L-amino acid sequence, and is composed of all D-amino acids (inverting the chirality of the α-center of the amino acid subunits) to help maintain side chain topology similar to that of the original L-amino acid peptide.

[0027] An “isolated” molecule is one that is in a form not found in nature, including those that have been purified.

[0028] The term “binding affinity” generally refers to the strength of the sum total of non-covalent interactions between a single binding site of a molecule and its binding partner (e.g., a receptor and its ligand, a Petition 870250092218, dated 09 / 10 / 2025, pp. 141 / 179 9 / 45 antibody and its antigen, two monomers that form a dimer, etc.). Unless otherwise indicated, as used in this document, “binding affinity” refers to the intrinsic binding affinity that reflects a 1:1 interaction between members of a binding pair. The affinity of a molecule X for its partner Y can generally be represented by the dissociation constant (Kd). Affinity can be measured by common methods known in the art, including those described in this document. Low-affinity antibodies generally bind to the antigen slowly and tend to dissociate easily, while high-affinity antibodies generally bind to the antigen more rapidly and tend to remain bound longer.

[0029] The affinity or avidity of a molecule for its binding partner can be determined experimentally using any suitable method known in the art, for example, flow cytometry, enzyme-linked immunosorbent assay (ELISA), or radioimmunoassay (RIA) or kinetics (e.g., KINEXA® or BIACORE™ or OCTET® analysis). Direct binding assays as well as competitive binding assay formats can be readily employed. (See, for example, Berzofsky et al., “Antibody-Antigen Interactions,” in Fundamental Immunology, Paul, W.E., ed., Raven Press: New York, NY (1984); Kuby, Immunology, W.H. Freeman and Company: New York, NY (1992)). The measured affinity of a particular binding pair interaction may vary if measured under different conditions (e.g., salt concentration, pH, temperature).Thus, affinity measurements and other antigen binding parameters (e.g., Kd or Kd, Kbound, Kunoff) are performed using standardized solutions of binding partners and a standardized buffer, as known in the art.

[0030] An “active agent” is an ingredient that is intended to provide biological activity. The active agent may be associated with one or more other ingredients. An active agent that is a peptide may also be called an “active peptide”.

[0031] An “effective quantity” of an active agent is a quantity Petition 870250092218, dated 09 / 10 / 2025, pp. 142 / 179 10 / 45 sufficient to accomplish a specifically stated purpose.

[0032] The term “pharmaceutical formulation” refers to a preparation that is in a form that allows the biological activity of the active ingredient to be effective and that does not contain additional components that are unacceptably toxic to a subject to whom the formulation would be administered. Such a composition may be sterile and may comprise a pharmaceutically acceptable vehicle, such as physiological saline. Suitable pharmaceutical compositions may comprise one or more of a buffer (e.g., acetate, phosphate or citrate buffer), a surfactant (e.g., polysorbate), a stabilizing agent (e.g., polyol or amino acid), a preservative (e.g., sodium benzoate) and / or other conventional solubilizing or dispersing agents.

[0033] A “subject” or “individual” or “animal” or “patient” or “mammal” is any subject, particularly a mammalian subject, for whom diagnosis, prognosis or therapy is desirable. Mammals include humans, domestic animals, farm animals, sporting animals and laboratory animals, including, for example, humans, non-human primates, canines, felines, swine, bovines, equines, rodents, including rats and mice, rabbits, etc.

[0034] Terms such as “treating” or “treatment” or “to treat” or “relieving” or “to alleviate” refer to therapeutic measures that cure, reduce, lessen the symptoms of, and / or halt the progression of a diagnosed pathological condition or disorder. In certain modalities, a subject is successfully “treated” for a disease or disorder if the patient experiences total, partial, or transient relief or elimination of at least one measurable symptom or physical parameter associated with the disease or disorder.

[0035] A “control patient” is a subject who has not received treatment of the invention. A “control population” or a “control patient population” is a group of subjects who have not received treatment of the invention. A control patient or subject in the control population has the Petition 870250092218, dated 09 / 10 / 2025, pp. 143 / 179 11 / 45 same disease or disorder as the subject being compared to the control patient or control population. For example, a clinical outcome of a cancer patient who received a pharmaceutical composition or method of the invention is compared to the average (median) outcome of subjects with the same type and / or stage of cancer who did not receive a pharmaceutical composition or method of the invention. In some embodiments, the control patient or patients in the control population received a treatment different from the treatment of the invention, for example, a standard care treatment.

[0036] An “antagonist” is a substance that prevents, blocks, inhibits, neutralizes, or reduces the biological activity or effect of another molecule, such as a receptor or ligand.

[0037] The terms “inhibit,” “block,” and “suppress” are used interchangeably and refer to any statistically significant decrease in occurrence or activity, including the complete blocking of occurrence or activity. For example, “inhibition” may refer to a decrease of approximately 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% in activity or occurrence. An “inhibitor” is a molecule, factor, or substance that produces a statistically significant decrease in the occurrence or activity of a process, pathway, or molecule.

[0038] A “neoplastic cell” or “neoplasm” has typically undergone some form of mutation / transformation, resulting in abnormal growth compared to normal cells or tissue of the same type. Neoplasms include morphological irregularities as well as pathological proliferation. Neoplastic cells can be benign or malignant. Malignant neoplasms, i.e., cancers, are distinguished from benign ones because they demonstrate loss of cell differentiation and orientation and have invasive and metastatic properties.

[0039] A “tumor” or “solid tumor” is a mass of neoplastic cells, such as cancerous cells. The terms “advanced,” “metastatic,” and “advanced / metastatic” are used interchangeably to describe a cancer. Petition 870250092218, dated 09 / 10 / 2025, pp. 144 / 179 12 / 45 in which malignant cells have migrated from the original tumor to another location, for example, another organ, in the patient's body.

[0040] “Pharmacokinetics” or “PK” refers to the study of how an administered substance is processed by a subject’s body. Pharmacokinetic determinations include how the substance enters the bloodstream (absorption), is dispersed or disseminated throughout the body’s fluids and tissues (distribution), is recognized and transformed by the body (metabolism), and is removed from the body (excretion). The substance can be a drug, for example, ST316. Pharmacokinetics can be assessed using various metrics, many of which are calculated based on the amount of the substance in the body (e.g., in plasma) at various points in time after administration of the substance.

[0041] Time after administration is measured from To, which is the moment when administration of a single dose of the substance is initiated. In cases where administration of the pharmaceutical composition is paused and resumed one or more times during the total infusion period, To is the start of the total infusion period.

[0042] The “total infusion duration” or “total infusion period” is the time from the start of a single dose of the pharmaceutical composition to the end of administration and includes infusion periods and interruption periods.

[0043] “Cmax” is a pharmacokinetic metric that refers to the peak plasma concentration of a substance after administration.

[0044] “Tmax” is a pharmacokinetic metric that refers to the time after the start of administration of the substance (To) to reach Cmax.

[0045] “Tultima” is a pharmacokinetic metric that refers to the time of the last quantifiable concentration of the substance.

[0046] “AUC” or “area under the curve” is a pharmacokinetic metric that describes the variation in the concentration of a substance in blood plasma as a function of time. AUC can be calculated for different time periods, for example, from time zero to a specified time t (AUCt or AUCo-t), from time Petition 870250092218, dated 09 / 10 / 2025, pp. 145 / 179 13 / 45 zero to infinity (AUC~ or AUCo-~), etc.

[0047] “Elimination half-life” or “half-life” or “t%” is a pharmacokinetic metric that refers to the time required for the concentration of a substance to reach half of its original value.

[0048] “Clearance” is a pharmacokinetic metric that refers to the volume of plasma cleared of the substance per unit of time.

[0049] “Vz” is a pharmacokinetic metric that refers to the volume of distribution during the terminal phase. II. Peptides and β-Catenin and Antagonist Peptides

[0050] The Wnt / β-catenin pathway is a validated target for cancer therapies. Constitutive activation of the Wnt / β-catenin signaling pathway is implicated in carcinogenesis, tumor progression, and poor prognosis of many malignancies (Shang 2017). The complex formed by the interaction between β-catenin and its coactivator, BCL9, drives oncogene expression in various cancer types due to aberrant Wnt pathway signaling. Disruption of the β-catenin / BCL9 complex has been shown to suppress oncogenic Wnt / β-catenin transcription without affecting the homeostatic functions of β-catenin (Takada 2012).

[0051] The interaction between β-catenin and BCL9 was previously considered an “indifferent” target due to the inability of small molecules to inhibit complex formation and the inability of antibodies to gain access to the cytoplasm or nucleus to disrupt the interaction (Takada 2012). Peptide antagonism of β-catenin differs from these other approaches because it can specifically target the interaction between β-catenin and BCL9 and is selectively toxic to tumor cells where these proteins are hyperactive, amplified, and / or mutated.

[0052] In some embodiments, the methods of the invention comprise treating a patient with a solid tumor with an effective amount of a β-catenin peptide antagonist. Preferably, the peptide antagonist Petition 870250092218, dated 09 / 10 / 2025, pp. 146 / 179 14 / 45 of β-catenin inhibits the interaction of β-catenin with BCL9. Peptide antagonists of β-catenin can be rationally designed, for example, based on the native sequence of BCL9 with modifications to improve the electrostatic interactions between the peptide antagonist and β-catenin. In particular, peptide antagonists of the interaction between β-catenin and BCL9 can be derived from the homology domain 2 (HD2) of BCL9 and designed to interact with the first armadillo repeat domain (ARM-1) of β-catenin, which is a site used by BCL9 but not by other β-catenin binding partners. Document no. WO 2021 / 007158 provides several examples of such peptides.

[0053] The ability of a wild-type BCL9-based peptide to antagonize β-catenin activity, such as inhibiting β-catenin interaction with BCL9, can be measured by methods described herein, for example, in Example 1. The cytotoxic activity of a β-catenin peptide antagonist can be measured in vitro by known assays and / or in vivo using known tumor models; for example, documents WO 2019 / 136125 and WO 2021 / 007158 describe such assays and models.

[0054] The β-catenin peptide antagonist can be a cell-penetrating peptide. In one embodiment, the peptide comprises a cell-penetrating domain. Several cell-penetrating peptide sequences are described and characterized in the literature (see document no. WO 2019 / 136125). In one embodiment, the peptide is a cyclic peptide. Cyclized peptides, for example, using hydrocarbon clamps (Bernal 2007; Bird 2017) or other cyclization methods known in the art, can enter cells via passive diffusion, endocytosis / endosomal escape, or other mechanisms (Dougherty 2019). Peptides can also be delivered to cells via mechanisms that exploit cellular receptors, for example, RGD-like sequences targeting integrin. Alternatively, peptides can be encapsulated and delivered to cells in vesicles, such as exosomes or liposomes, or in micelles. Petition 870250092218, dated 09 / 10 / 2025, pages 147 / 179 15 / 45

[0055] The β-catenin peptide antagonist may have a modified N-terminus and / or a modified C-terminus. For example, peptide antagonists may optionally include an N-terminal acetyl group and / or a C-terminal amide group. Other examples of optional N-terminal and / or C-terminal groups include hydrophobic groups, such as a linear or cyclic C2C18 aliphatic or aromatic hydrocarbon, a naphthyl group, a phenyl group, an octanoyl group, and a valeryl group, including an isovaleryl group. In some embodiments, the peptide antagonist comprises a linker or spacer between the peptide and the hydrophobic group. Such linkers or spacers include, for example, aminohexanoic acid, beta-alanine, substituted alkyls, substituted cycloalkyls, and polyethylene glycol.

[0056] ST316 is a novel 25-amino acid peptide antagonist of the β-catenin and BCL9 interaction. ST316 is composed entirely of D-amino acids and is highly stable in the presence of proteolytic enzymes. Due to its resistance to protease degradation, ST316 has a long plasma half-life and should not be processed and presented by antigen-presenting cells, thus avoiding the stimulation of an anti-drug antibody (ADA) response.

[0057] ST316 is composed of two domains: (i) a 15-amino acid N-terminal active domain derived from the homology domain 2 (HD2) of BCL9 and designed to interact with the first armadillo repeat domain (ARM-1) of β-catenin, which is a site used by BCL9 but not by other β-catenin binding partners; and (ii) a 10-amino acid C-terminal domain, which increases solubility and cell penetration. The D amino acid sequence of ST316 is: FRWLLRQLARLAQLAKLTPVKLTPV (SEQ ID NO: 1). The N-terminal active domain is in bold and the C-terminal cell penetration domain is in italics. ST316 also comprises an N-terminal octanoyl group.

[0058] ST316 represents the first well-tolerated approach to drugging the Wnt pathway by specifically binding to the n-terminal of β-catenin, where it interacts with BCL9, a cofactor that is not required for normal physiological function, Petition 870250092218, dated 09 / 10 / 2025, pages 148 / 179 16 / 45 but it is necessary for the pathological, oncogenic and immunosuppressive functions of β-catenin. Compositions and Administration

[0059] In certain aspects, the invention provides a composition, for example, a pharmaceutical composition, comprising a β-catenin peptide antagonist, such as ST316. The pharmaceutical composition is suitable for parenteral administration. In one embodiment, ST316 may be in salt form. Preferably, the composition comprises one or more carriers, diluents, excipients, or other additives. For example, the composition may comprise one or more bulking agents, one or more buffers, one or more pH adjusting agents, and / or one or more diluents.

[0060] Aspects of the invention relate to methods of administering a β-catenin peptide antagonist to a subject. The β-catenin peptide antagonist is administered parenterally. Parenteral routes of administration include intravenous (IV), intramuscular, intraperitoneal, intrathecal, and subcutaneous routes. In a preferred embodiment, a pharmaceutical composition comprising ST316 is administered by IV infusion. The pharmaceutical composition may be provided, for example, in an IV fluid comprising normal saline (0.9%), half-normal saline (0.45%), or 5% dextrose in water (D5W). In one embodiment, the β-catenin peptide antagonist is a sterile, lyophilized solid presented in vials for reconstitution, for example, with Sterile Water for Injection USP. The reconstituted peptide may subsequently be diluted for intravenous administration.

[0061] The β-catenin peptide antagonist can be dosed based on the patient's weight. The β-catenin peptide antagonist, such as ST316, can be administered to a patient at a dose of approximately 0.25 mg / kg to approximately 12 mg / kg. In certain embodiments, ST316 is administered at a dose of approximately 0.25 mg / kg, approximately 0.5 mg / kg, approximately 0.75 mg / kg, approximately 1 mg / kg, approximately 1.5 mg / kg, approximately 2 mg / kg, approximately 2.5 mg / kg, approximately 3 mg / kg, Petition 870250092218, dated 09 / 10 / 2025, pages 149 / 179 17 / 45 approximately 3.5 mg / kg, approximately 4 mg / kg, approximately 4.5 mg / kg, approximately 5 mg / kg, approximately 5.5 mg / kg, approximately 6 mg / kg, approximately 6.5 mg / kg, approximately 7 mg / kg, approximately 7.5 mg / kg, approximately 8 mg / kg, approximately 9 mg / kg, approximately 10 mg / kg, approximately 11 mg / kg or approximately 12 mg / kg. These amounts can also serve as endpoints for a range of doses to be administered, for example, approximately 0.75 mg / kg to approximately 7 mg / kg, approximately 2 mg / kg to approximately 4 mg / kg, etc.

[0062] Alternatively, the β-catenin peptide antagonist may be administered at a fixed dose. The β-catenin peptide antagonist, such as ST316, may be administered to a patient at a dose of approximately 500 mg to approximately 1500 mg. In certain embodiments, ST316 is administered at a dose of approximately 500 mg, approximately 550 mg, approximately 600 mg, approximately 650 mg, approximately 700 mg, approximately 750 mg, approximately 800 mg, approximately 850 mg, approximately 900 mg, approximately 950 mg, approximately 1000 mg, approximately 1100 mg, approximately 1200 mg, approximately 1300 mg, or approximately 1400 mg or approximately 1500 mg. These amounts can also serve as endpoints for a range of doses to be administered, for example, approximately 600 ng to approximately 1100 mg, approximately 750 mg to approximately 900 mg, etc.

[0063] In embodiments of the invention, the pharmaceutical composition is administered to the subject by intravenous infusion, in which the total duration of the infusion does not exceed about 360 minutes. In some embodiments, the total duration of the infusion is from about 30 minutes to about 240 minutes. For example, the total duration of the infusion may be 30 minutes, 60 minutes, 90 minutes, 120 minutes, 150 minutes or 180 minutes, or an intermediate duration, such as 45 minutes, 100 minutes, etc. In certain embodiments, the total duration of the infusion is from about 60 minutes to about 90 minutes, or from about 60 minutes to about 120 minutes, or from about 90 minutes to about 120 minutes, or from about 60 minutes to about 180 minutes.

[0064] In some modalities, the infusion of the pharmaceutical composition may be interrupted, that is, it may be temporarily interrupted and then Petition 870250092218, dated 09 / 10 / 2025, pp. 150 / 179 18 / 45 resumption. The duration of the interruption may vary, for example, it may be approximately 15 minutes or less, or approximately 30 minutes or less, or approximately one hour or less, or approximately two hours or less, or approximately three hours or less, or approximately four hours or less.

[0065] In some embodiments, the pharmaceutical composition may be administered with one or more secondary agents intended to block histamine release, prevent or improve infusion-related reactions (IRRs), reduce fever or inflammation, and / or relieve itching and / or hives. IRRs may include, for example, erythema, fever, chills / shivering, tachycardia, tachypnea, hypotension, and bronchospasm. One or more secondary agents may be administered concomitantly with, before, and / or after administration of the pharmaceutical composition. One or more secondary agents may be administered in a separate composition from the pharmaceutical composition or may be combined with the pharmaceutical composition. In addition, one or more secondary agents may be administered by the same route as the pharmaceutical composition, or may be administered by a different route (e.g., orally).

[0066] Examples of one or more secondary agents for administration with the pharmaceutical composition include, but are not limited to, antihistamines, including H1 antagonists, H2 antagonists and mast cell degranulation inhibitors (e.g., acrivastine, astemizole, azatadine, azelastine, bepotastine, brompheniramine, burfroline, cetirizine, chlorzoxazone, chlorpheniramine, cromoglycate, cyproheptadine, desloratadine, dexbrompheniramine, diphenhydramine, doxantrozole, epinastine, etodroxyzine, famotidine, fexofenadine, forskolin, hydroxyzine, isoproterenol, ketotifen, levocetirizine, loratadine, lodoxamide, mequitazine, methyllazine, mizolastine, nedocromil, olopatadine, oxatomide, pemirolast, pimecrolimus, pirbuterol, pizotifen, proxicromil, ranitidine, terfenadine, terbutaline); leukotriene inhibitors (e.g., montelukast, zafirlukast, zileuton); nonsteroidal anti-inflammatory drugs (NSAIDs) (e.g., ibuprofen, naproxen, aspirin); Petition 870250092218, dated 09 / 10 / 2025, pp. 151 / 179 19 / 45 acetaminophen / paracetamol; corticosteroids (e.g., hydrocortisone, dexamethasone, prednisone, prednisolone); anti-nausea medications (e.g., prochlorperazine, ondansetron); and saline solution and / or electrolytes. In a preferred embodiment, the secondary agent is an antihistamine, such as chlorpheniramine or diphenhydramine. In specific embodiments, the secondary agent is selected from the group consisting of acetaminophen / paracetamol, an H1 antagonist and an H2 antagonist, montelukast, an antiemetic, and combinations thereof.

[0067] In certain embodiments, the secondary agent is administered during the administration of the pharmaceutical composition, such as during the infusion period. In certain embodiments, the secondary agent, such as an antihistamine, is administered to the subject prior to the administration of the pharmaceutical composition, such as approximately 7 days prior, or approximately 6 days prior, or approximately 5 days prior, or approximately 4 days prior, or approximately 72 hours prior, or approximately 48 hours prior, or approximately 24 hours prior, or approximately 8-12 hours prior, or approximately 68 hours prior, or approximately 4-6 hours prior, or approximately 2-4 hours prior, or approximately 1-2 hours prior, or approximately 1 hour prior, or immediately prior. In some embodiments, the secondary agent is administered within 24 hours after the administration of the pharmaceutical composition.For example, the secondary agent may be administered immediately after, approximately 0.5-1 hour after, approximately 1-2 hours after, approximately 2-4 hours after, approximately 4-6 hours after, approximately 6-8 hours after, approximately 8-12 hours after, or approximately 24 hours after the completion of administration of the pharmaceutical composition.

[0068] The secondary agent may be administered several times before, during, and / or after administration of the pharmaceutical composition. Combinations of secondary agents may be administered simultaneously or at different times. For example, an antihistamine may be administered before administration of the pharmaceutical composition, and a corticosteroid may be administered after administration of the pharmaceutical composition.

[0069] The peptide antagonist is normally administered to the patient Petition 870250092218, dated 09 / 10 / 2025, pp. 152 / 179 20 / 45 several times. The β-catenin peptide antagonist may be administered once a week, once every two weeks, once every three weeks, once every four weeks, or combinations of these intervals. The total duration of treatment may be at least one, two, three, four, five, six, seven, eight, nine, ten, eleven, or twelve weeks, or at least three, four, five, six, seven, eight, nine, ten, eleven, or twelve months.

[0070] In one embodiment, administration of the β-catenin peptide antagonist may occur once weekly for a duration of at least one week, two weeks, or three weeks (i.e., one, two, or three administrations), six weeks (i.e., six administrations), nine weeks, twelve weeks, three months, six months, nine months, or twelve months. In another embodiment, administration may occur once every two weeks for a duration of at least four weeks (i.e., two administrations), eight weeks (i.e., four administrations), twelve weeks, three months, six months, nine months, or twelve months.In some modalities, a patient may receive the β-catenin peptide antagonist once a week for a period of at least one week, two weeks, three weeks, six weeks, nine weeks, twelve weeks, three months, six months, nine months, or twelve months, followed by administration once every two weeks for at least four weeks, eight weeks, twelve weeks, three months, six months, nine months, or twelve months.

[0071] Neoadjuvant therapy may be administered to reduce the size or extent of a tumor, or to make subsequent treatment easier, more effective, or less extensive. In one particular modality, a β-catenin peptide antagonist may be administered as a neoadjuvant before, for example, surgery and / or radiation, as a single agent or in combination with a secondary agent.

[0072] The pharmacokinetics (PK) and pharmacodynamics of the β-catenin antagonist can be evaluated by standard methods and as described in the Examples. In some embodiments, the β-catenin antagonist is ST316. Petition 870250092218, dated 09 / 10 / 2025, pp. 153 / 179 21 / 45 III. Treatment Methods

[0073] Subjects requiring treatment by the methods of the invention are patients diagnosed with a solid tumor. For example, the subject may have an advanced, unresectable, and metastatic solid tumor. In one embodiment, the tumor harbors one or more abnormalities in the Wnt / β-catenin signaling pathway. By way of non-limiting example, one or more abnormalities in the Wnt / β-catenin signaling pathway may comprise one or more mutations in one or more adenomatous polyposis coli (APC), APC membrane recruitment protein 1 (AMER1), Axin2, β-catenin, RING finger protein 43 (RNF43), and / or T-cell factor 7 (TCF7).

[0074] In some embodiments, the subject has locally advanced or metastatic breast cancer (BC), colorectal cancer (CRC), cholangiocarcinoma (CCA), endometrial cancer, hepatocellular carcinoma (HCC), melanoma, non-small cell lung cancer (NSCLC), ovarian cancer (OC), pancreatic adenocarcinoma (PDAC), or synovial sarcoma (SS). In certain embodiments, the disease is refractory or intolerant to other available therapies that would impact survival. In particular embodiments of the invention, administration of ST316 may inhibit tumor growth, reduce tumor volume, or a combination thereof.

[0075] In one modality, the patient was diagnosed with BC. More than 50% of BC tumors have been shown to exhibit abnormalities in Wnt signaling (Lin 2000). Triple-negative breast cancer (TNBC) is a particularly aggressive form of BC that generally has a worse prognosis compared to other BC subtypes and accounts for 10-15% of all BC cases (Won 2020). Patients with TNBC do not respond to hormonal and targeted therapies due to a lack of hormone receptors and human epidermal growth factor receptor 2 (HER2). Chemotherapy remains the standard of care, despite its limited benefits. Therefore, the medical need for more effective agents in this clinical setting remains very high. In TNBC, mutations in the Wnt pathway are known to drive Petition 870250092218, dated 09 / 10 / 2025, pp. 154 / 179 22 / 45 tumorigenesis and metastasis lead to poor clinical outcomes and increase the likelihood of developing pulmonary and cerebral metastases (Pohl 2017).

[0076] In one modality, the patient was diagnosed with CRC. CRC is one of the most frequently diagnosed cancers worldwide, ranking second in women and third in men globally (Sung 2021). Approximately 95% of patients with metastatic CRC (mCRC) have microsatellite stable disease (MSS) and are not eligible for treatment with immune checkpoint inhibitors. This patient population relies on modestly effective chemotherapy and targeted therapy regimens, making it an area of ​​significant unmet medical need. Over 90% of CRC tumors have been shown to exhibit abnormalities in Wnt signaling. These have been shown to drive disease initiation, progression, and metastasis (Nie 2020).

[0077] In one instance, the patient was diagnosed with CCA. Also known as biliary tract cancer, CCA refers to a heterogeneous group of gastrointestinal cancers arising from the bile ducts, gallbladder, or ampulla of Vater. These cancers are often difficult to diagnose due to their anatomical location and the scarcity or nonspecificity of their symptoms; therefore, >75% of patients with CCA present with advanced and unresectable disease (Lamarca 2014, Takahashi 2013). Advanced unresectable CCA represents an area of ​​unmet medical need due to its highly aggressive nature, limited treatment options, and poor prognosis, especially after first-line treatment, usually with a combination of gemcitabine and cisplatin (Valle 2010). Clinical and preclinical studies have suggested the key role of Wnt / ε-catenin signaling pathway activation in the induction and progression of CCA (Zhang 2020).

[0078] In one modality, the patient was diagnosed with OC. OC is the fifth leading cause of cancer death in women and the leading cause of death among gynecologic cancers (ACS 2022). Two-thirds of patients with OC are diagnosed at stages 3 and 4, representing a rate of Petition 870250092218, dated 09 / 10 / 2025, pages 155 / 179 23 / 45 relatively low 5-year survival rates of 39% and 17% for tumors in stages 3 and 4, respectively (NCCN 2022). Advanced OC represents an area of ​​unmet medical need due to its highly aggressive nature, limited treatment options, and poor prognosis. Wnt activity has been shown to correlate with histological grade, epithelial-to-mesenchymal transition, chemotherapy resistance, and poor prognosis in OC (Teeuwssen 2019).

[0079] In one modality, the patient was diagnosed with endometrial cancer. Endometrial cancer is the most common gynecological cancer in the industrialized world and is increasing in women of all ages, at least in part due to the increased incidence of obesity and resulting hyperinsulinemia (Moore 2017; Parrish 2022). Older women have a higher risk of recurrence and higher mortality rates (Moore 2017). Mutations in β-catenin are associated with worse outcomes in patients with endometrial cancer, including a higher incidence of recurrence and lower survival rates (Parrish 2022).

[0080] The effectiveness of treatment can be evaluated by one or more known measures. For example, patients undergoing the methods of the invention may exhibit results including prolonged survival, improved progression-free survival, improved duration of response, longer remission, reduced risk of relapse and / or improved tumor response to treatment with a β-catenin peptide antagonist, compared with the same result(s) in patients not undergoing the methods of the invention, i.e., control patients. A result in a patient treated by a method of the invention can be compared, for example, to the median result in a population of control patients. The population of control patients may be administered, for example, with a regimen selected from the group consisting of placebo, surgery, radiation, chemotherapy, immunotherapy, hormonal therapy, targeted therapy and combinations thereof.Comparisons can be analyzed statistically using, for example, rank tests. Petition 870250092218, dated 09 / 10 / 2025, pp. 156 / 179 24 / 45 signaled Wilcoxon or the Kaplan-Meier method.

[0081] In one embodiment, the outcome in a patient receiving a β-catenin peptide antagonist, such as ST316, optionally in combination with standard treatment, is compared with the median outcome in control patients receiving a placebo. In another embodiment, the outcome in a patient receiving a β-catenin peptide antagonist, such as ST316, is compared with the median outcome in control patients receiving standard treatment.

[0082] Tumor response to treatment can be assessed, for example, by measuring tumor burden and / or tumor regression. Treatment response compares one or more efficacy measures after a treatment regimen compared to baseline, for example, before treatment with ST316. An initial assessment is preferably performed within 24, 48, or 72 hours, or within 1, 2, 3, or 4 weeks before the first treatment with a β-catenin peptide antagonist. In a preferred modality, a baseline assessment is performed within one week before the first treatment with ST316.

[0083] “Tumor burden” is the total mass or total size of cancerous tissue in a patient’s body. Tumor response can be assessed by measures that include objective response rate, including partial and / or complete response, stable disease, disease control rate, duration of disease control, and duration of response. These parameters can be determined, for example, by the Revised Response Assessment Criteria in Solid Tumors (RECIST 1.1) (Eisenhauer 2009).

[0084] The objective response rate assesses the reduction in tumor size, for example, tumor diameter, which can be determined by clinical examination and / or imaging. When a patient has multiple tumors, the tumor size can optionally be expressed as the average diameter of all tumors or as the sum of the diameters of all tumors. Superficial tumors can be measured clinically, for example, using calipers or by photography and Petition 870250092218, dated 09 / 10 / 2025, pp. 157 / 179 25 / 45 ruler measurement. Imaging methods include computed tomography (CT), typically with contrast; X-ray; magnetic resonance imaging (MRI); and positron emission tomography (PET), such as (18)Ffluorodeoxyglucose PET. In a preferred modality, CT is used to assess tumor response, for example, in a patient with BC or a patient with melanoma. Consequently, in one aspect, the invention provides a method for reducing tumor burden, i.e., tumor mass and / or tumor size, in a patient, the method comprising administering to the patient a β-catenin peptide antagonist, such as ST316. The reduction in tumor burden is measured relative to baseline.

[0085] In certain modalities, particularly those in which the assessment is made by RECIST 1.1, the disease control rate defines the tumor response level as the best of the following: complete response (CR), which is the disappearance of the tumor(s); partial response (PR), which is a decrease of at least 30% in the size of the tumor(s); stable disease (SD), in which the change in tumor size is decreased by less than 30% or increased by less than 20%; or disease progression, which is an increase of at least 20% in tumor size and / or new lesions. Patients treated by the methods of the invention may present CR, PR or SD.

[0086] The duration of disease control is the period of time from achieving a response (CR or PR) or DS until disease progression. The duration of response is the period of time from achieving a response until disease progression, i.e., the period during which a tumor does not grow or spread, or until death. The duration of response in patients receiving treatment with ST316 may be, for example, at least 4, 6, 8, 10, or 12 weeks, at least 4, 6, 8, 10, 12, 16, 18, or 24 months, or at least 3, 4, or 5 years. Patients treated by the methods of the invention may experience a longer duration of disease control or a longer duration of response. Consequently, in one aspect, the invention provides a method for increasing the duration of disease control or the duration of response in Petition 870250092218, dated 09 / 10 / 2025, pp. 158 / 179 26 / 45 a patient, the method comprising administering to the patient a β-catenin peptide antagonist, such as ST316. The increase in duration of disease control or duration of response is measured relative to the median duration of disease control or duration of response, respectively, in a control population.

[0087] Survival can be assessed as overall survival, i.e., the time a patient lives, as progression-free survival, i.e., the time a patient is treated without progression or worsening of the disease, or as event-free survival, i.e., the time a patient remains free of complications or negative events, such as relapse or disease progression. Survival begins from the date treatment starts. Overall survival, median overall survival, progression-free survival, median progression-free survival, event-free survival, and median event-free survival can be calculated, for example, by Kaplan-Meier analysis, based on the response to treatment.

[0088] Consequently, in one aspect, the invention provides a method for increasing overall survival in a patient, the method comprising administering to the patient a β-catenin peptide antagonist, such as ST316. The increase in overall survival can be measured relative to the median overall survival in a control population. Alternatively, an increase compared to a control population in the percentage of patients surviving for a given period of time (e.g., 6 months or 12 months) indicates an increase in overall survival.

[0089] In another aspect, the invention provides a method for increasing progression-free survival in a patient, the method comprising administering to the patient a β-catenin peptide antagonist, such as ST316. The increase in progression-free survival can be measured relative to the median progression-free survival in a control population. Alternatively, an increase compared to a control population in the percentage of patients who are relapse-free for a given period of time. Petition 870250092218, dated 09 / 10 / 2025, pp. 159 / 179 A 27 / 45 time frame (e.g., 6 months or 12 months) indicates an increase in progression-free survival.

[0090] In a further aspect, the invention provides a method for increasing event-free survival in a patient, the method comprising administering to the patient a β-catenin peptide antagonist, such as ST316. The increase in event-free survival is measured relative to the median event-free survival in a control population. Alternatively, an increase compared to a control population in the percentage of patients who are event-free for a given period of time (e.g., 6 months or 12 months) indicates an increase in event-free survival.

[0091] A patient is successfully treated according to the methods of the invention if the patient exhibits or exhibits at least one of the following outcomes after administration of a β-316 peptide antagonist, such as ST316: - undetectable tumor (or at least one tumor, if multiple tumors are present at baseline); - at least approximately a 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95% reduction in tumor size compared to baseline; - no significant increase (e.g., less than 20% or less than 30%) in tumor size compared to baseline; - Significantly increased duration of response, optionally compared to the median duration of response in a control patient population; - a significant increase in the duration of disease control, optionally compared to the median duration of disease control in a control patient population; - significant increase in progression-free survival, optionally compared to the median progression-free survival of a Petition 870250092218, dated 09 / 10 / 2025, pages 160 / 179 28 / 45 control patient population; - Significant increase in overall survival, optionally compared to the median overall survival of a control patient population. IV. Preparation Methods

[0092] Peptide antagonists of β-catenin can be synthesized chemically, for example, using solid-phase peptide synthesis or solution-phase peptide synthesis, or a combination of both. Synthesis can optionally occur as peptide fragments that are subsequently combined chemically or enzymatically.

[0093] Alternatively, β-catenin peptide antagonists can be expressed using recombinant methods. For example, nucleic acid molecules encoding ST316 can be constructed by chemical synthesis using an oligonucleotide synthesizer. Nucleic acid molecules can be designed based on the amino acid sequence of ST316 and the selection of those codons that are favored in the host cell in which recombinant ST316 will be produced. Standard methods can be applied to synthesize a nucleic acid molecule encoding a β-catenin peptide antagonist, such as ST316.

[0094] Once prepared, the nucleic acid encoding the peptide can be inserted into an expression vector and operatively ligated to an appropriate expression control sequence for peptide expression in a desired host. In order to obtain high levels of peptide expression, the nucleic acid must be operatively ligated or associated with transcription and translation expression control sequences that are functional in the chosen expression host.

[0095] A wide variety of host / expression vector combinations can be employed, any known in the art. Useful expression vectors include, for example, vectors comprising expression control sequences of SV40, bovine papillomavirus, adenovirus and Petition 870250092218, dated 09 / 10 / 2025, pp. 161 / 179 29 / 45 cytomegalovirus. Useful expression vectors for bacterial hosts include known bacterial plasmids, such as E. coli plasmids, including pCR1, pBR322, pMB9 and their derivatives, plasmids of wider host variation such as M13, and filamentous single-stranded DNA phages.

[0096] Suitable host cells include prokaryotes, yeast, insect, or higher eukaryotic cells under the control of appropriate promoters. Prokaryotes include Gram-negative or Gram-positive organisms, for example E. coli or bacilli. Higher eukaryotic cells may be established or mammalian-derived cell lines, examples of which include Pichia pastoris, 293 cells, COS-7 cells, L cells, C127 cells, 3T3 cells, Chinese hamster ovary (CHO) cells, HeLa cells, and BHK cells. Cell-free translation systems may also be employed.

[0097] Peptides can be purified using methods that include, for example, reversed-phase high-performance liquid chromatography (RP-HPLC), multicolumn countercurrent solvent gradient purification (MCSGP), hydrophobic interaction chromatography (HIC), and ion-exchange chromatography. EXAMPLES

[0098] The embodiments of the present disclosure may be best defined by reference to the following non-limiting examples. It will be evident to those skilled in the art that many modifications, both to the materials and to the methods, may be made without departing from the scope of the present disclosure. Example 1. ST316 binds to β-catenin and disrupts its interaction with BCL9.

[0099] We used a fluorescence polarization assay to measure the binding between FITC-conjugated ST316 and recombinant β-catenin. Increasing concentrations of full-length β-catenin (0.24 nM–2000 nM) were incubated with 12.5 nM ST316-FITC for one hour at room temperature. Fluorescence polarization was measured using a SpectraMax® plate reader. ST316 binds to Arm1 β-catenin with a Kd of approximately Petition 870250092218, dated 09 / 10 / 2025, pp. 162 / 179 30 / 45 126.1 nM (Figure 1). The binding affinity of BCL9 and β-catenin is approximately 200 nM, indicating that ST316 binds β-catenin with an affinity equal to or greater than that of the endogenous ligand.

[0100] We used an ALPHA assay to quantify the interaction of BCL9 and β-catenin. The ALPHA assay was performed in an assay buffer composed of 20 mM MES (pH 6.5), 60 mM NaCl, 0.01% BSA, 1 mM DTT, 0.05% Tween-20. ST316 was serially diluted to a final concentration ranging from 50 μM to 6 nM and added to the plate, followed by the addition of 25 nM of full-length recombinant β-catenin. The plate was incubated for 1 minute at room temperature. A 26-amino acid C-terminal biotinylated BCL9 HD-2 peptide was added to each well at a final concentration of 50 nM, followed by the addition of a rabbit polyclonal anti-β-catenin antibody (Ab1605) (final concentration of 50 ng / ml).Donor and acceptor beads (PerkinElmer AlphaScreen Streptavidin Donor Beads 6760007 and Protein A Acceptor Beads 6760136, respectively) were added to the mixture at a final concentration of 10 μg / ml each, and the plate was protected from light and incubated for one hour at room temperature on a plate shaker. The alpha signal was then measured using a SpectraMax® plate reader. The results showed that both the ST316 peptide and HD2 inhibit the interaction of BCL9 and β-catenin with an IC50 of approximately 1 μM (Figure 2). Example 2. In Vitro Effects of ST316

[0101] We examined the effect of ST316 on the nuclear localization of β-catenin in HCT116 cells. Briefly, cells were cultured in T25 culture flasks with McCoy's 5A medium (with 10% FBS and Mycozap) and treated with vehicle only, 3 μM ST316, or 5 μM ST316 for 24 hours. Cells were fractionated by first harvesting followed by incubation in lysis buffer with DTT and a protease inhibitor cocktail. Then, a 10 IGEPAL CA-630 solution was added to the cells. The cells were centrifuged to collect the supernatant cytoplasm fraction. The remaining pellet was resuspended in extraction buffer containing DTT and a protease inhibitor cocktail. The cells Petition 870250092218, dated 09 / 10 / 2025, pp. 163 / 179 31 / 45 were centrifuged and the nuclear supernatant was collected and analyzed by Western blot (Figure 3A). Treatment with ST316 5 μM significantly reduced the nuclear localization of β-catenin. Figure 3B indicates the percentage of nuclear and cytoplasmic β-catenin in the Western blot image, as quantified by the ImageJ analysis software.

[0102] To examine the effect of ST316 on BCL9 proteasomal degradation, Colo320 colorectal cancer cells were incubated with ST316 for 24 hours, with or without the proteasome inhibitor MG132. No impact on cell viability was observed. Protein expression was analyzed using the Jess Protein Detection System. The results are shown in Figure 4A-4D. Similar results were observed in HCT116 cells. Example 3. ST316 exhibits an inhibitory effect on the invasion of cancer cells in vitro.

[0103] We used an in vitro Boyden chamber assay to investigate the inhibitory effect of ST316 on neoplastic cell invasion. HCT116 colorectal cancer cells (2 x 10⁵) were equilibrated in serum-free cell culture medium for two hours in a Matrigel-coated invasion plate insert, after which complete growth medium plus 10% fetal bovine serum was added to the bottom well of each insert as a chemoattractant. Cells were then exposed to 1 μM ST316 or negative control peptide for 24 hours. No significant impact on cell viability was observed under these conditions.

[0104] For analysis, the medium contained in the inserts was aspirated and the top layer of the membrane was removed. After fixation with MeOH and staining with crystal violet, the membranes were removed and mounted on a glass slide. Image acquisition of the cell-invaded membranes was performed using a Nikon Eclipse T2S microscope at 20x magnification and a Nikon Digital Sight 1000 microscope. Cell counts were determined manually using ImageJ. The data represent the mean and standard error of three biological replicates containing at least three fields of Petition 870250092218, dated 09 / 10 / 2025, pp. 164 / 179 32 / 45 representative views. The results are shown in Figure 5A-5B. Example 4. ST316 Administration Inhibits Tumor Growth in Vivo

[0105] The effect of ST316 on tumor formation in APCmin / + mice, which harbor an APC mutation necessary for the regulation of β-catenin concentration, was examined. APCmin / + mice are predisposed to cachexia and develop intestinal adenoma. Six-week-old female APCmin / + mice received ST316 at a dose of 10 mg / kg once weekly by subcutaneous injection (n=3 per group). The mice received a total of 10 doses. Body weight was measured three times a week during the study. After week 16, the mice were euthanized, plasma was collected for cytokine analysis, and the small intestine was collected for adenoma quantification.Treatment with ST316 resulted in a 55% reduction in adenoma formation (Figure 6A), prevented the onset of cachexia (Figure 6B), and increased serum expression of the pro-inflammatory cytokine CCL2 / CCL4 (Figure 6C), compared to mice that did not receive ST316 treatment.

[0106] Furthermore, we examined the effect of ST316 on tumor volume in a subcutaneous tumor model using colorectal cancer cells. Briefly, HCT116 cells (5 x 105), suspended 1:1 in Matrigel, were implanted via subcutaneous injection into the axilla of female NOD / SCID mice. ST316 was administered at a dose of 5 mg / kg by subcutaneous injection three times a week for three weeks. Dosing was initiated on day 12 after tumor inoculation, with a mean initial tumor volume of approximately 200 mm3. Tumor volume was monitored three times a week. Treatment with ST316 resulted in 99% inhibition of tumor growth compared to vehicle or a control peptide (Figure 7).

[0107] The effect of ST316 on tumor volume was also examined in a subcutaneous tumor model using triple-negative breast cancer cells. Briefly, 4T1-luc cells (5 x 105), suspended 1:1 in Matrigel, were Petition 870250092218, dated 09 / 10 / 2025, pages 165 / 179 33 / 45 tumors were implanted via subcutaneous injection in the axilla of female NOD / SCID mice. ST316 was administered at a dose of 5 mg / kg by subcutaneous injection once weekly for eight weeks. Dosing was initiated on day 6 after tumor inoculation, with a mean initial tumor volume of approximately 100 mm3. Tumor volume was monitored three times weekly. Treatment with ST316 resulted in 84% inhibition of tumor growth compared to a control peptide (Figure 8A). Tumors were collected on day 60 and analyzed for Axin2 expression by quantitative PCR (n=3 per group). The results are shown in Figure 8B. Example 5. ST316 is safe and well tolerated in animal models. Safety Pharmacology

[0108] The potential pharmacological effects of ST316 on the cardiovascular (mini-pigs), respiratory (rats and mini-pigs), and central nervous system (CNS) (rats and mini-pigs) were investigated in 28 / 29-day GLP toxicology studies. No ST316-related effects were observed on body weight, food consumption, ophthalmological examinations, respiratory function, or CNS in rats or mini-pigs. Furthermore, in rats, there were no ST316-related effects on the Functional Observational Battery (FOB), and in mini-pigs, there were no ST316-related effects on electrocardiogram (ECG) rhythm, morphology, or quantitative measurements for QRS, RR, PR, QT, or QTcF intervals.

[0109] ADA formation was not detected in a non-GLP toxicology study in C57BL / 6 mice after weekly dosing for 10 weeks or in a GLP toxicology study in minipigs after weekly dosing for 4 weeks (5 doses), and was within the 5% assay-qualified error rate in a GLP toxicology study in rats after weekly dosing for 4 weeks (5 doses). One positive sample in the GLP minipig study (96 samples total) and 2 positive samples (1%) at the 10 mg / kg dose level in the GLP rat study from a control animal not exposed to ST316 (198 samples total) were observed. Petition 870250092218, dated 09 / 10 / 2025, pp. 166 / 179 34 / 45 Tolerability

[0110] The findings in the GLP toxicology studies were mainly limited to the injection site in rats and minipigs. None of the observations definitively related to ST316 in animals that received ST316 via catheter were considered adverse. In minipigs, no pathological findings related to the test article were observed. Assessment of pseudoallergic reactions

[0111] DLT associated with ST316 exposure in MTD studies was a dose-dependent infusion-related pseudoallergic reaction (IRR) observed in mice, rats, and minipigs. Clinical signs of pseudoallergic reaction in mice may include decreased body temperature, reduced activity, and piloerection. Non-clinical toxicology of IV-administered ST316 was characterized in a 29-day GLP toxicology study in rats and a 28-day GLP toxicology study in minipigs, which were supported by 7-day maximum tolerated dose (MTD) and repeated dose range (RDF) determination studies in the same species.

[0112] Toxicological studies to characterize the mechanism of ST316 pseudoallergic IRR have demonstrated that the ST316 reaction is mediated by mast cell activation and histamine release, and is attenuated by pretreatment with antihistamines (ranitidine and pyrilamine) and / or the leukotriene receptor antagonist (montelukast). Example 6: Administration of ST316 to patients with solid tumors

[0113] An open-label, two-part, phase 1-2 study is being conducted to determine the safety, tolerability, pharmacokinetics (PC), pharmacodynamics, and proof-of-concept efficacy of intravenously administered ST316 in selected subjects with advanced solid tumors likely to have abnormalities in the Wnt / β-catenin signaling pathway. Abnormalities in the Wnt / β-catenin pathway are an appropriate target due to their impact on carcinogenesis, tumor progression, and prognosis of tumor types. Petition 870250092218, dated 09 / 10 / 2025, pp. 167 / 179 35 / 45 of tumors selected for this study, as well as their impact on the immunological constituents of the tumor microenvironment. Furthermore, non-clinical studies have demonstrated specific ST316 antagonism of β-catenin and potent in vitro / in vivo activity in breast and colon cancer models. This study utilizes a novel peptide drug with a novel mechanism of action in subjects who have exhausted standard treatments. The study consists of two phases: a phase 1 dose-escalation / regimen exploration phase and a phase 2 expansion phase. Dose escalation phase

[0114] The dose-escalation phase employs a standard approach to assess safety, tolerability, and select a recommended phase 2 dose in a wide range of advanced unresectable and metastatic solid tumors, including subjects diagnosed with locally advanced or metastatic BC, CCA, CRC, endometrial cancer, HCC, melanoma, NSCLC, OC, PDAC, or SS, whose disease is refractory or intolerant to all available therapies that would impact survival. Table 1 shows the dose-escalation phase study design. Table 1. Overview of dose escalation in phase 1. ^ Population (est. n = 25) • Types of recurrent / refractory solid tumors with known abnormalities of the Wnt / β-catenin signaling pathway ^ Objectives • Understand safety, PK, PD • Refine dosage ^ Design • 3 + 3 • Monotherapy • 1x / weekly IV o 12 mg / kg (N=3-6) o 8 mg / kg (N=3-4) o 4 mg / kg (N=4) Petition 870250092218, dated 09 / 10 / 2025, pages 168 / 179 36 / 45 o 2 mg / kg (N=3) o 1 mg / kg (N=4) o 0.5 mg / kg (N=3)

[0115] ST316 is administered intravenously. Dose-escalation cohorts are recruited using a standard 3+3 design. The duration of the initial dose infusion is 60-180 minutes for all subjects and may be modified in subsequent doses based, at least in part, on the presence or absence of IRRs.

[0116] Dose cohorts are 0.5, 1.2, 4, 8, and 12 mg / kg once weekly (QW), optionally with dose cohorts of 0.25, 0.75, 1.5, 3, and 6 mg / kg and fixed dosing. One treatment cycle consists of 21 days QW.

[0117] Subjects continue treatment until disease progression, withdrawal of consent, or lack of clinical benefit, as determined by the treating physician. Patients may continue treatment beyond progression if they are clinically stable. If repeat imaging shows a reduction or stabilization in tumor burden compared to the initial examination demonstrating PD, treatment is continued as scheduled. If repeat imaging confirms PD, therapy is discontinued for patients. In determining whether tumor burden has increased, stabilized, or decreased, researchers consider all target lesions as well as non-target lesions.

[0118] The National Cancer Institute Common Terminological Criteria for Adverse Events (CTCAE), v5.0, are used to classify toxicities. Safety assessments include adverse events, serious adverse events, physical examinations, vital sign measurements, Eastern Cooperative Oncology Group (ECOG) status, clinical safety laboratory assessments (hematology, serum chemistry and liver panels, coagulation and urinalysis), and electrocardiograms. All subjects are followed for safety issues for at least 30 days after the last dose of ST316. Petition 870250092218, dated 09 / 10 / 2025, pages 169 / 179 37 / 45

[0119] To mitigate potential IRRs, subjects are pre-treated prior to the first dose of ST316 with (i) daily montelukast (10 mg PO) starting at least 2 days before ST316 administration and on the day of administration; and (ii) both an H1 and an H2 antagonist on the day of ST316 dosing, at least 30 minutes before the start of the infusion. If oral antihistamines are used, they will be administered earlier based on predicted Cmax. Preferred H1 / H2 antagonists include famotidine (20 mg PO or 20-40 mg IV) plus chlorpheniramine (10 mg IV / PO) or diphenhydramine (50 mg PO / IV).

[0120] Subjects are monitored for IRRs for 24 hours after the first administration of ST316, for 4 hours after the 2nd infusion, and for 2 hours after subsequent infusions. Treatment of IRRs includes, for example, pausing and decreasing the infusion rate and / or administering antihistamines such as diphenhydramine, leukotriene receptor antagonists, paracetamol, nonsteroidal anti-inflammatory drugs (NSAIDs), IV fluids, antiemetics, oxygen, bronchodilators, and, if other measures are inadequate, corticosteroids. Table 2 provides non-limiting examples of agents for the treatment of IRRs. Table 2. Agents for treating IRR Agent Dose Route Acetaminophen 1000 mg IV or PO Diphenhydramine 25-50 mg IV or PO Ranitidine 50 mg IV or PO Ibuprofen 200-400 mg PO Naproxen 500 mg PO 0.9% saline solution 1L or PRN IV (warmed) Prochlorperazine 5-10 mg IV or PO Ondansetron 4-16 mg IV or PO

[0121] Safety data available, as well as PK, PD and data Efficacy factors are considered when selecting the maximum tolerated dose and the recommended dose for phase 2, and to potentially model the dose cohort(s). Petition 870250092218, dated 09 / 10 / 2025, pages 170 / 179 38 / 45 fixed, if applicable. Once the decision on the recommended dose and regimen for phase 2 has been made, the study proceeds to the expansion phase. Effectiveness

[0122] Patients from Cohorts 1-4 are receiving treatment and have undergone an initial assessment, as summarized in Table 3. The results show two patients with stable CRC, with one patient exceeding 4 months and one patient exceeding 6 months. Table 3. Summary of dose escalation for phase 1 Cohort Dose (mg / kg) Cancer Type Enrollment Date Status 1 0.5 NSCLC June 2023 Progressed 1 0.5 NSCLC June 2023 SD (18 weeks); Out of study 1 0.5 CRC July 2023 Progressed 2 1 PDAC August 2023 Progressed 2 1 TNBC September 2023 Progressed 2 1 NSCLC September 2023 Progressed 2 1 CRC September 2023 SD (27+ weeks); In study 3 2 CRC November 2023 SD (18+ weeks); Under study 3 2 CRC November 2023 Progressed 3 2 CRC December 2023 Progressed 4 4 PDAC January 2024 Progressed 4 4 CRC January 2024 Under Study 4 4 CRC February 2024 Under Study 4 4 CRC February 2024 Under Study Security

[0123] The Wnt pathway, which ultimately signals via β-catenin, plays an important role in healthy individuals. β-catenin is an active transcription factor that controls certain stem cell populations and organ regeneration. Given that the Wnt pathway is also critical for many types Petition 870250092218, dated 09 / 10 / 2025, pp. 171 / 179 In cases of cancer (39 / 45), there have been many clinical attempts to medicate this pathway, despite concerns about "targeting outside the tissue." Previous attempts include: pan-Wnt inhibitors, which suffer from toxicity directed at gastrointestinal stem cells and bones (Kahn 2014; Shaw 2019); inhibition of Wnt pathway members such as Frizzled, Tankyrase, Dickkopf-1, and Porcupine; and disruption of the β-catenin transcription complex by targeting T-cell factor / lymphoid enhancing factor. All of these approaches have resulted in toxicity.

[0124] Significantly, safety data from Cohorts 1-4 do not reveal dose-limiting toxicities, infusion-related reactions, and serious adverse events related to ST316. Pharmacokinetics and Pharmacodynamics

[0125] Pharmacokinetic (PK) data from the first four patient cohorts show relatively linear PK parameters, with no significant accumulation or increase in clearance over time. The AUC(o-i68) for Cohort 2 exceeded the predicted limit for biological activity from the model. The results are shown in Table 4 and Figures 9A-9B. Table 4. Pharmacokinetic Summary Dose (mg / kg) Infusion Duration Cmax (pg / ml) AUC(o-i68) (h* pg / ml 0.5 1.5 h 8.8 159.1 1 1.5 h 15.3 237.7 2 1.5 h 30.1 461.6 4 1.5 h 66.7 1085

[0126] Blood samples collected from patients before and after exposure to ST316 were analyzed for immune cell populations and show a clear pharmacodynamic effect of ST316 on the WNT-catenin pathway. Treatment with ST316 resulted in a reduction in the population of immunosuppressive polymorphonuclear myeloid-derived (PMN) suppressor cells (Figures 10A-10C). PMN-MDSCs exhibit a CD3 phenotype. Petition 870250092218, dated 09 / 10 / 2025, pp. 172 / 179 40 / 45 CD19-CD14-HLADRlowCD11b+CD15+.

[0127] Taken together, the clinical data demonstrate that the administered doses of ST316 are pharmacological and that ST316 is safe, reaches its target, and positively affects the disease in patients. Expansion Phase

[0128] The expansion phase consists of three colorectal cancer cohorts. Fifteen subjects will be included in Cohort 1 with ST316 monotherapy. If one or more subjects present a positive efficacy criterion defined as stable disease (SD) for more than four months; or any response, partial response (PD) or complete response (CR), 15 more patients will be recruited to Cohort 1. If Cohort 1 is safe, Cohorts 2 and 3 will be recruited.

[0129] Cohort 2 will recruit 15 patients in the second line to be treated with ST 316 in combination with bevacizumab and FOLFIRI. If two or more subjects show a response (PR or CR) and / or PFS for more than seven months, additional subjects may be recruited.

[0130] Cohort 3 will recruit 15 patients with up to four prior lines of therapy to be treated with ST 316 in combination with fruquintinib. If one or more subjects show a response (PR or CR) and / or PFS for more than four months, additional subjects may be recruited.

[0131] Additional tumor types (e.g., endometrial cancer, melanoma, TNBC, ovarian cancer) and combinations may be added in the expansion phase, based on efficacy signals during the dose escalation and expansion phases. Pharmacokinetic Assessment

[0132] The plasma PK of ST316 is assessed using standard methods. A complete plasma PK profile, including AUCt, Cmax, t½, AUC- and tmax, is obtained and analyzed using non-compartmental methods for all subjects in the dose escalation phase. Blood samples are collected before dosing, at the end of the infusion, and at various time points after the infusion during the first cycle. Petition 870250092218, dated 09 / 10 / 2025, pp. 173 / 179 41 / 45 and less frequently thereafter. Pharmacodynamic Assessment

[0133] Blood and tumor samples are collected for pharmacodynamic evaluation, which may include analysis of circulating tumor DNA for mechanistically associated genetic abnormalities and changes over time, and tumor analysis at baseline and during treatment using quantitative reverse transcriptase polymerase chain reaction, ribonucleic acid sequencing, NanoString analysis, and immunohistochemistry.

[0134] Relevant disease markers, such as tumor biomarkers, are collected at screening and during the study. At screening, all subjects have an excisional or core biopsy obtained after the last dose of prior systemic therapy and before recruitment. Post-treatment biopsy of the same lesion is performed, if possible, and if the tumor persists in subjects who had a screening biopsy. Effectiveness assessment

[0135] RECIST 1.1 (Eisenhauer 2009) is used to evaluate tumor response, disease control rate (DCR), duration of response (DOR), and / or progression-free survival (PFS). The time between the first administration of study treatment and the first documented disease progression or death determines the PFS. The time between the first observed response and the first documented disease progression or death determines the DOR.

[0136] CT or computed MRI are used to assess tumor response, with CT being the preferred imaging technique. All baseline and study assessments use the same methodology. All affected organs are documented as target or non-target lesions at the start of the study, according to RECIST 1.1, and followed throughout the study.

[0137] A radiological assessment of a complete response (CR) or partial response (PR) requires confirmatory imaging at least 4 weeks after the initial assessment of the response has been observed. Significant clinical benefit may be used instead of a definite response. Petition 870250092218, dated 09 / 10 / 2025, pages 174 / 179 42 / 45 radiologically to expand expansion cohorts. REFERENCES American Cancer Society (ACS). Key Statistics for Ovarian Cancer. Web page accessed November 16, 2022. Bernal F, et al. Reactivation of the p53 Tumor Suppressor Pathway by a Stapled p53 Peptide. J. Am. Chem. Soc. 2007; 129:2456-2457. Bird GH, et al. Biophysical Determinants for Cellular Uptake of HydrocarbonStapled Peptide Helices. Nat. Chem. Biol. 2017; 12:845-852. Bugter JM, et al. Mutations and mechanisms of WNT pathway tumour suppressors in cancer. Nat Rev Cancer. Janeiro de 2021;21(1):5-21. de la Roche M, et al. The function of BCL9 in Wnt / beta-catenin signaling and colorectal cancer cells. BMC Cancer. 15 de julho de 2008;8:199. Dotan E, et al. Phase Ib Study of Wnt Inhibitor Ipafricept with Gemcitabine and nab-paclitaxel in Patients with Previously Untreated Stage IV Pancreatic Cancer. Clin Cancer Res. 15 de outubro de 2020;26(20):5348-5357. Dougherty PG, et al. Understanding Cell Penetration of Cyclic Peptides. Chem. Rev. 2019; 119(17):10241-10287. Eisenhauer EA, et al. New response evaluation criteria in solid tumours: Revised RECIST guideline (versão 1.1). Eur J Cancer 2009; 45(2):228-247. Gay DM, et al. Loss of BCL9 / 9l suppresses Wnt driven tumourigenesis in models that recapitulate human cancer. Nat Commun. 13 de fevereiro de 2019;10(1):723. Petição 870250092218, de 09 / 10 / 2025, pág. 175 / 179 43 / 45 Kahn M, Can we safely target the WNT pathway? Nat. Rev. Drug Discov. julho de 2014;13(7):513-532. Lamarca A, et al. Second-line chemotherapy in advanced biliary cancer: a systematic review. Annals of oncology. 1 de dezembro de 2014;25(12):2328-38. doi: 10.1093 / annonc / mdu162. Lin SY, et al. Beta-catenin, a novel prognostic marker for breast cancer: its roles in cyclin D1 expression and cancer progression. Proc Natl Acad Sci U S A. 11 de abril de 2000;97(8):4262-6. Liu J, et al. Wnt / e-catenin signalling: function, biological mechanisms, and therapeutic opportunities. Signal Transduct Target Ther. 3 de janeiro de 2022;7(1):3. Mieszczanek J, et al. Bcl9 and Pygo synergise downstream of Apc to effect intestinal neoplasia in FAP mouse models. Nat Commun. 13 de fevereiro de 2019;10(1):724. Moor AE, et al. BCL9 / 9L-e-catenin Signaling is Associated With Poor Outcome in Colorectal Cancer. EBioMedicine. 30 de outubro de 2015;2(12):1932-43. Moore K, et al. Endometrial Cancer: Is This a New Disease? Am. Soc. Clin. Oncol. Educational Book. 2017 435-442. National Comprehensive Cancer Network (NCCN) Clinical Practice Guidelines in Oncology (NCCN guidelines) with NCCN Evidence Blocks - Ovarian Cancer 2022. Nie X, et al. Emerging roles of Wnt ligands in human colorectal cancer. Frontiers Petição 870250092218, de 09 / 10 / 2025, pág. 176 / 179 44 / 45 in oncology. 14 de agosto de 2020;10:1341. doi: 10.3389 / fonc.2020.01341 Parrish ML, et al. Mechanisms of mutant β-catenin in endometrial cancer progression. Frontiers Oncol. 29 de setembro de 2022;12:1009345. Pohl, SG, et al. Wnt signaling in triple-negative breast cancer. Oncogenesis. 2017; 6:e310. doi: 10.1038 / oncsis.2017.14. Ruiz de Galarreta M, et al. β-Catenin Activation Promotes Immune Escape and Resistance to Anti-PD-1 Therapy in Hepatocellular Carcinoma. Cancer Discov 2019;9(8):1124-41. doi: 10.1158 / 2159-8290.CD-19-0074. Shang S, et al. The regulation of β-catenin activity and function in cancer: therapeutic opportunities. Oncotarget 2017;8(20):33972-33989. Shaw HV, et al. Targeting the Wnt signalling pathway in cancer: prospects and perils. Swiss Medical Weekly 2019;149:w20129. Simon R. Optimal two-stage designs for phase 2 clinical trials. Controlled Clinical Trials 1989;10:1-10. Sung H, et al. Global Cancer Statistics 2020: GLOBOCAN Estimates of Incidence and Mortality Worldwide for 36 Cancers in 185 Countries. CA Cancer J Clin. maio de 2021;71(3):209-249. doi: 10.3322 / caac.21660. Epub 4 de fevereiro de 2021. PMID: 33538338. Takada K, et al. Targeted disruption of the BCL9 / e-catenin complex inhibits oncogenic Wnt signaling. Sci Transl Med. 22 de agosto de 2012;4(148). Takahashi Y, et al. Surgical treatment of perihilar cholangiocarcinoma in Petição 870250092218, de 09 / 10 / 2025, pág. 177 / 179 45 / 45 octogenarians: a single center experience. J Hepatobiliary Pancreat Sci. março de 2013;20(3):324-31. Teeuwssen M, et al. Wnt signaling in ovarian cancer stemness, EMT, and therapy resistance. Journal of clinical medicine. 11 de outubro de 2019;8(10):1658. Vafaizadeh V, et al. The interactions of Bcl9 / Bcl9L with β-catenin and Pygopus promote breast cancer growth, invasion, and metastasis. Oncogene. outubro de 2021;40(43):6195-6209. Valle J, et al. Cisplatin plus gemcitabine versus gemcitabine for biliary tract cancer. N Engl J Med. 8 de abril de 2010;362(14):1273-81. doi: 10.1056 / NEJMoa0908721. Wang J, et al. Low BCL9 expression inhibited ovarian epithelial malignant tumor progression by decreasing proliferation, migration, and increasing apoptosis to cancer cells. Cancer Cell Int. 6 de dezembro de 2019;19:330. Won KA, et al. Triple-negative breast cancer therapy: Current and future perspectives (Review). Int J Oncol. dezembro de 2020;57(6):1245-1261. Zhang GF, et al. Wnt / e-catenin signaling as an emerging potential key pharmacological target in cholangiocarcinoma. Bioscience Reports. 27 de março de 2020;40(3). Zhong Y, et al. Tankyrase Inhibition Causes Reversible Intestinal Toxicity in Mice with a Therapeutic Index < 1. Toxicol Pathol. February 2016;44(2):267-78. *** The present invention is further described by the following claims. Petition 870250092218, dated 09 / 10 / 2025, pp. 178 / 179

Claims

1 / 3 CLAIMS 1. A method for treating a solid tumor in a patient, wherein the method is characterized in that it comprises administering parenterally to the patient a pharmaceutical composition comprising an effective amount of a β-catenin peptide antagonist.

2. Method, according to claim 1, characterized in that the solid tumor is selected from the group consisting of breast cancer, colorectal cancer, cholangiocarcinoma, endometrial cancer, hepatocellular carcinoma, melanoma, non-small cell lung cancer, ovarian cancer, pancreatic adenocarcinoma and synovial sarcoma.

3. A method according to claim 1, characterized in that the patient has received prior treatment selected from a group consisting of chemotherapy, hormone therapy, radiation, targeted therapy, and combinations thereof.

4. Method according to claim 1, characterized in that the peptide antagonist is administered as a neoadjuvant.

5. Method according to claim 1, characterized in that the peptide antagonist comprises the amino acid sequence D FRWLLRQLARLAQLAKLTPVKLTPV (SEQ ID NO: 1).

6. Method according to claim 1, characterized in that the peptide antagonist is administered to the patient at a dose of approximately 0.25-12 mg / kg.

7. Method of administering a β-catenin peptide antagonist to a subject, wherein the method is characterized in that it comprises parenteral administration to the subject of a pharmaceutical composition comprising the peptide antagonist, wherein the peptide antagonist comprises the amino acid sequence D FRWLLRQLARLAQLAKLTPVKLTPV (SEQ ID NO: 1), and wherein the peptide antagonist is administered at a dose of approximately 0.25-12 mg / kg.

8. Method according to claim 1, characterized in that the peptide antagonist is administered at a dose selected from about 0.5 mg / kg, about 1 mg / kg, about 2 mg / kg, about 4 mg / kg, about 8 mg / kg and about 12 mg / kg.

9. Method according to claim 1, characterized in that the peptide antagonist disrupts the binding of β-catenin to the B-cell CLL / lymphoma 9 (BCL9) protein.

10. Method according to claim 1, characterized in that the peptide antagonist comprises an N-terminal octanoyl group.

11. Method according to claim 1, characterized in that the peptide antagonist is ST316.

12. Method according to claim 1, characterized in that the pharmaceutical composition is administered intravenously.

13. Method or composition, as defined in claim 12, characterized in that the pharmaceutical composition is administered by means of infusion.

14. Method or composition, according to claim 13, characterized in that the infusion duration is approximately 30-180 minutes.

15. Method or composition, according to claim 14, characterized in that the infusion duration is about 60-90 minutes.

16. Method according to claim 1, characterized in that the pharmaceutical composition is administered once a week.

17. Method according to claim 1, characterized in that the pharmaceutical composition is administered once every two weeks.

18. Method according to claim 1, characterized in that the pharmaceutical composition is administered for at least four weeks.

19. Method according to claim 1, characterized in that it further comprises administering to the patient bevacizumab and folinic acid, Petition 870250092218, dated 09 / 10 / 2025, page 121 / 179 3 / 3 fluorouracil and irinotecan (FOLFIRI).

20. Method according to claim 1, characterized in that it further comprises administering fruquintinib to the patient. Petition 870250092218, dated 09 / 10 / 2025, pp. 122 / 179

Images