Anti-cancer TGFB2 drugs with immunotherapy

Abstract

This invention relates to a method for treating or relieving cancer symptoms in human or animal subjects using a pharmaceutical composition designed to enhance antitumor effects. An exemplary synergistic drug therapy includes a composition having a combination of active agents, comprising an agent for inhibiting or suppressing TGF-β2 expression, an immune checkpoint inhibitor, and an active interleukin immunotherapy agent. One or more biomarkers may be used to select subjects that would benefit from this method. TIFF2026519788000011.tif144128

Description

Technical Field

[0001] Sequence Listing This application includes a sequence listing that was created on May 15, 2024, and electronically submitted as a ST.26 file named 018988-012WO1_SL.xml, with a size of 120,193 bytes.

[0002] Technical Field The present invention relates to agents, compositions, and methods for use in treating or alleviating the symptoms of cancer. Exemplary combinations of synergistic therapies include active agents for inhibiting or suppressing the expression of TGF-β2, immune checkpoint inhibitors, and interleukin immunotherapeutic agents. One or more biomarkers can be used to select subjects who will benefit from the agents, compositions, and methods.

Background Art

[0003] Background Cancer is a complex pathological condition involving multiple diverse cellular pathways. Due to this complexity, many anticancer drugs have limited or partial therapeutic efficacy.

[0004] Disadvantages of conventional therapies include a lack of efficacy, judged by overall survival.

[0005] Further disadvantages of conventional therapies include serious unwanted side effects such as killing healthy cells in addition to cancer cells.

[0006] Additional disadvantages of anticancer agents include high toxicity at the required therapeutic dosing levels.

[0007] There is a need for agents, compositions, and methods for cancer diseases to increase efficacy and reduce toxicity and unwanted side effects.

[0008] To provide significant antitumor and cancer immunotherapy effects, therapeutic compositions of different agents are needed, which can improve efficacy, reduce side effects, and mitigate adverse health effects. Improved guidance is needed regarding the use of such compositions, including the use of appropriate biomarkers to select synergistic effects between the agents and compositions.

[0009] To treat various cancers, there is an urgent need for new methods, agents, and uses that combine strategies for cancer immunotherapy with strategies for direct antitumor attacks. These therapies include therapeutic compositions that combine cancer T-cell therapy and immunotherapy with potent anticancer agents. [Overview of the project]

[0010] overview This invention provides agents, compositions, and methods for use in treating or relieving the symptoms of cancer. These compositions can enhance antitumor effects across a variety of different cancers. The synergistic pharmacotherapy of this invention involves using potent direct antitumor agents in combination with cancer immunotherapy agents. Modalities of cancer immunotherapy in this disclosure include combination therapies with immune checkpoint inhibitors and protein immunotherapies. The agents, compositions, and methods of this invention can combine strategies for cancer immunotherapy with strategies for direct antitumor attacks to treat a variety of cancers.

[0011] In some embodiments, the compositions and methods of the present invention can increase efficacy and reduce toxic side effects and adverse health effects in the treatment of cancer.

[0012] In further embodiments, the methods and therapeutic strategies of the present invention may include enhanced guidance for improving patient outcomes by using appropriate biomarkers to select synergistic effects of compositions.

[0013] Exemplary synergistic drug therapies include combinations of active agents, such as agents for inhibiting or suppressing TGF-β2 expression, immune checkpoint inhibitors, and interleukin immunotherapies.

[0014] In a further context, one or more biomarkers, including IRF5 and ITGAM, can be used to select subjects who will benefit from the composition and therapeutic method.

[0015] In certain embodiments, the composition and treatment method can be applied in combination with chemotherapy and other standard treatments for cancer.

[0016] The embodiments of the present invention include the following:

[0017] A method for treating or relieving the symptoms of cancer in a subject in need, A step of administering a composition containing an agent for inhibiting or suppressing the expression of TGF-β2; The process of administering an immune checkpoint inhibitor; and The process of administering interleukin immunotherapy agents. Methods that include...

[0018] Agents for inhibiting or suppressing TGF-β2 expression, in combination with immune checkpoint inhibitors and active interleukin immunotherapies, for use in treating or relieving cancer symptoms.

[0019] A composition comprising an agent for inhibiting or suppressing TGF-β2 expression for use in the preparation of pharmaceuticals, or for treating or relieving cancer symptoms in a subject, in combination with an immune checkpoint inhibitor and an active interleukin immunotherapy agent, and a pharmaceutically acceptable carrier.

[0020] The method, agent, or composition described above, wherein the cancer is a solid tumor, pancreatic cancer, melanoma, lung cancer, breast cancer, multiple myeloma, or colorectal cancer.

[0021] The method, agent, or composition as described above, wherein the agent for inhibiting or suppressing the expression of TGF-β2, the immune checkpoint inhibitor, and the active interleukin immunotherapeutic agent are administered together, simultaneously, sequentially, or at separate time points.

[0022] The method, agent, or composition as described above, wherein the composition and agent are administered by infusion or injection.

[0023] The method, agent, or composition as described above, wherein the agent for inhibiting or suppressing the expression of TGF-β2 is selected from Table 1 or Table 2, and their chemically modified variants, their LNA variants, their gapmer variants, and any combination or pool thereof.

[0024] The agent for inhibiting or suppressing the expression of TGF-β2 is TIFF2026519788000002.tif12128 in the method, agent, or composition as described above.

[0025] The method, agent, or composition as described above, wherein the agent or composition comprises a carrier that is sterile water for injection, physiological saline, isotonic saline, or a combination thereof.

[0026] The method, agent, or composition as described above, wherein the agent or composition is substantially excipient-free.

[0027] The method, agent, or composition as described above, wherein the composition is stable in the carrier at 37°C for at least 14 days.

[0028] The method, agent, or composition as described above, wherein the immune checkpoint inhibitor is an inhibitor of PD-1, CTLA-4, or PD-L1.

[0029] The method, agent, or composition described above, wherein the immune checkpoint inhibitor is pembrolizumab, nivolumab, semiprimab, spartalizumab, atezolizumab, avelumab, or durvalumab.

[0030] The method, agent, or composition described above, wherein the active interleukin immunotherapy agent is natural IL-2, high-dose IL-2, recombinant IL-2, or aldesleukin.

[0031] The above-described method, agent, or composition, wherein, upon administration or use, the subject has reduced TGF-β2 expression.

[0032] The above-mentioned method, agent, or composition, comprising using one or more biomarkers to select subjects who will benefit from the method, agent, or use.

[0033] The method, agent, or composition described above, wherein one or more of the biomarkers are IRF5 levels, ITGAM levels, or a combination thereof.

[0034] The method, agent, or composition described above, wherein the subject is selected when one or more of the aforementioned biomarkers are ITGAM, and the expression of ITGAM is at a level exceeding that seen in healthy patients.

[0035] The above method, agent, or composition wherein, after the administration or use, the subject has an increased level of IRF5.

[0036] The above method, agent, or composition wherein, after the administration or use, the subject has an increased level of ITGAM.

[0037] The method, agent, or composition described above, comprising administering to the subject a therapeutically sufficient amount of a pharmaceutical composition containing an expression product which is IRF5 or ITGAM.

[0038] The method, agent, or composition described above, wherein the expression product is mRNA, polypeptide, protein, or a fragment thereof, or a combination thereof.

[0039] The above-mentioned method, agent, or composition, wherein the administration or use reduces mortality at 6, 12, 18, 24, 30, or 36 months.

[0040] The above-mentioned method, agent, or composition, wherein the administration or use increases the survival rate at 6, 12, 18, 24, 30, or 36 months.

[0041] The above-mentioned method, agent, or composition wherein the administration or use of the composition is combined with a standard treatment for cancer, the standard treatment comprising chemotherapy or radiotherapy. [Brief explanation of the drawing]

[0042] [Figure 1] Figure 1 shows the Kaplan-Meier overall survival chart (KM plotter) obtained in a study of clinical outcomes in melanoma patients. Figure 1 shows that the use of PD-1 checkpoint inhibitors in selected patients with high ITGAM was associated with improved survival in the case of low TGF-β2 (log-rank P=0.0039). [Figure 2] Figure 2 shows the Kaplan-Meier overall survival chart (KM plotter) obtained in a study of clinical outcomes in pancreatic cancer. Figure 2 shows that improved survival was demonstrated in the case of high IL-2 use in selected patients with high ITGAM (log-rank P=0.034). [Figure 3] Figure 3 shows the Kaplan-Meier overall survival chart obtained in a study of clinical outcomes for melanoma (KM plotter). Figure 3 shows that the use of PD-1 checkpoint inhibitors in selected patients with high IRF5 was associated with improved survival in the case of low TGF-β2 (log-rank P = 0.00053). [Figure 4]Figure 4 shows the Kaplan-Meier overall survival chart (KM plotter) obtained from a study of clinical outcomes in cancer patients. This study included multiple types of tumors. Figure 4 shows that improved survival was observed when the ratio of IL-2 to TGF-β2 (IL-2 / TGF-β2) was high with the use of PD-1 checkpoint inhibitors (log-rank P = 0.0031). [Figure 5] Figure 5 shows the Kaplan-Meier overall survival chart obtained in a study of the clinical outcomes of melanoma (KM plotter). Figure 5 shows that improved survival was observed when the ratio of IL-2 to TGF-β2 (IL-2 / TGF-β2) was high with the use of PD-1 checkpoint inhibitors (log-rank P = 3.7e-06). [Figure 6] Figure 6 shows the Kaplan-Meier overall survival chart (KM plotter) obtained from a study of clinical outcomes in melanoma patients. Figure 6 shows that improved survival was observed in cases of high IL-2 with regard to CKI use (log-rank P=0.0015). [Figure 7] Figure 7 shows the Kaplan-Meier overall survival chart (KM plotter) obtained in a study of clinical outcomes in melanoma patients. Figure 7 shows that improved survival was observed in patients with low TGF-β2 levels with the use of CKI (log-rank P=0.006). [Figure 8] Figure 8 shows the Kaplan-Meier overall survival chart obtained in a study of clinical outcomes in cancer patients (KM plotter). Figure 8 shows that improved survival was observed in cases of high IL-2 with the use of PD-1 checkpoint inhibitors (log-rank P=0.007). [Figure 9] Figure 9 shows the Kaplan-Meier overall survival chart obtained in a study of clinical outcomes in cancer patients (KM plotter). Figure 9 shows that improved survival was demonstrated in the case of low TGF-β2 with the use of PD-1 checkpoint inhibitors (log-rank P=0.0028). [Figure 10]Figure 10 shows the Kaplan-Meier overall survival chart (KM plotter) obtained from a study of clinical outcomes in melanoma patients. This study included multiple tumor types. Figure 10 shows that improved survival was observed with respect to the use of CKIs, with a higher ratio of IL-2 to TGF-β2 (IL-2 / TGF-β2) (log-rank P = 3.7e-06). [Figure 11] Figure 11 shows the Kaplan-Meier overall survival chart obtained in a study of clinical outcomes in cancer patients (KM plotter). Figure 11 shows that improved survival was observed when the ratio of IL-2 to TGF-β2 (IL-2 / TGF-β2) was high with regard to the use of PD-1 checkpoint inhibitors (log-rank P = 2.2e-08). [Modes for carrying out the invention]

[0043] Detailed explanation of this disclosure This invention relates to methods, compositions, and uses thereof for treating or relieving cancer symptoms in human or animal subjects using pharmaceutical compositions designed to promote antitumor effects across various different cancers.

[0044] Examples of synergistic drug therapies include compositions that are various combinations of active agents, including agents for inhibiting or suppressing TGF-β2 expression, checkpoint inhibitors, and interleukin immunotherapies.

[0045] In some embodiments, one or more biomarkers, including IRF5 and / or ITGAM, may be used to select subjects who will benefit from the method, agent, or use. The composition may be used in combination with chemotherapy and other standard therapeutic therapies.

[0046] The patients or subjects requiring cancer therapies described herein may be human or animal subjects.

[0047] As used herein, the term "agent" can refer to one or more active compounds; a combination of active compounds; or a composition containing one or more active compounds with a carrier and / or a solvent and / or any number of excipients. In some embodiments, such composition may be a pharmaceutical composition. In certain embodiments, such composition may be a pharmaceutical composition containing a therapeutically effective amount of one or more active compounds. The formulation of the active agent may be determined by those skilled in the art. Some examples of excipients are shown in Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, Pa. 1975, and Liberman, HA and Lachman, L., Eds., Pharmaceutical Dosage Forms, Marcel Decker, New York, NY, 1980. Methods for determining the therapeutically effective amount of a compound are known in the art.

[0048] Anti-cancer methods and compositions Aspects of the present invention include combinations of TGF-β2-specific antisense agents, IL-2 immunotherapy agents, and checkpoint inhibitors.

[0049] In certain embodiments, the present invention provides a therapeutic combination of one or more TGF-β2-specific antisense agents, IL-2 immunotherapeutic agents, and PD-1 checkpoint inhibitors.

[0050] In some cases, the therapeutic combinations of the present invention can be used in the treatment or remission of cancer symptoms.

[0051] In a further context, the therapeutic combination of the present invention can be used in the treatment or remission of symptoms of solid tumors.

[0052] In certain contexts, the therapeutic combinations of the present invention can be used in the treatment or remission of symptoms of solid tumors, breast cancer, pancreatic cancer, melanoma, lung cancer, multiple myeloma, and colorectal cancer.

[0053] In certain contexts, the therapeutic combinations of the present invention can provide cytotoxic effects that offer antitumor effects. For example, they can delay tumor growth.

[0054] Aspects of the present invention provide therapeutic combinations that have a significant effect on the survival of cancer patients. In some aspects, tumor levels of TGF-β2, IL-2, or the ratio IL-2 / TGF-β2 can have a significant impact on the overall survival of patients treated with immune checkpoint inhibitors.

[0055] In certain embodiments, the checkpoint inhibitor may be a PD-1 checkpoint inhibitor.

[0056] In some aspects, embodiments of the present invention can result in remarkably increased IL2 / TGF-β2 ratios and improved survival in patients treated with checkpoint inhibitors. The increase in IL2 / TGF-β2 ratios and improved survival may be due to a therapeutic combination of TGF-β2-specific antisense agents, IL-2 immunotherapeutic agents, and checkpoint inhibitors. In certain embodiments, the checkpoint inhibitor may be a PD-1 checkpoint inhibitor.

[0057] Aspects of the present invention aim to utilize a therapeutic combination of a TGF-β2-specific antisense agent, an IL-2 immunotherapy agent, and a checkpoint inhibitor as a powerful driver of survival for cancer patients.

[0058] In additional embodiments, therapeutic combinations of TGF-β2-specific antisense agents, IL-2 immunotherapies, and checkpoint inhibitors described herein can more than double overall survival for multiple types of cancer and tumors.

[0059] In further embodiments, therapeutic combinations of TGF-β2-specific antisense agents, IL-2 immunotherapies, and checkpoint inhibitors described herein can be used to increase the number of long-term survivors for multiple types of cancer and tumors.

[0060] Aspects of the present invention demonstrate that when a TGF-β2-specific antisense agent is used in combination with a checkpoint inhibitor and an interleukin immunotherapy agent in the treatment or remission of cancer symptoms, a synergistic effect can be demonstrated through a remarkable increase in anticancer efficacy.

[0061] In certain embodiments, a combination of an antisense agent for inhibiting or suppressing TGF-β2 expression, a PD-1 checkpoint inhibitor, and an IL-2 immunotherapy agent can be used to observe remarkable synergistic anticancer efficacy in treating or relieving cancer symptoms. This combination may be particularly effective in patients with high levels of tumor-associated monocytes and / or tumor-associated macrophages.

[0062] While we do not wish to be bound by theory, the combination of agents that inhibit or suppress TGF-β2 expression with PD-1 checkpoint inhibitors and IL-2 immunotherapies showed synergistic effects on high levels of tumor-associated monocytes and macrophages. This synergistic effect may be strongest in combination with PD-1 checkpoint inhibitors. Since PD-1 is present in M2-type tumor-associated macrophages, and antisense agents that inhibit or suppress TGF-β2 expression are effective in repolarizing M2 and promoting antitumor effects, it is thought that these two agents can function synergistically against the same target when used in combination with IL-2.

[0063] While we do not wish to be bound by theory, TGF-β2 plays a central role in programming M1 tumor-associated macrophages that can exhibit antitumor effects. The inhibitory or suppression of TGF-β2 by antisense agents has been demonstrated herein by the inventors to have antitumor effects. Antisense agents may have a reprogramming effect that enhances M1 tumor-associated macrophages. Such reprogramming, in particular when combined with agents inhibited by high levels of TGF-β2, may have the effect of actively reducing and / or eliminating cancerous tumors.

[0064] The present invention Administer to the subject a composition containing an agent for inhibiting or suppressing TGF-β2 expression in a therapeutically sufficient amount; Administering a therapeutically sufficient amount of a composition containing a checkpoint inhibitor to the subject; and Administer to the subject a therapeutically sufficient amount of a composition containing an interleukin immunotherapy agent. This includes methods for treating or relieving cancer symptoms in human or animal subjects in need.

[0065] Further aspects of the present invention include the following steps: Administer to the subject a composition containing an agent for inhibiting or suppressing TGF-β2 expression in a therapeutically sufficient amount; Administering to the subject a therapeutically sufficient amount of a composition containing an immune checkpoint inhibitor; and Administer to the subject a therapeutically sufficient amount of a composition containing an interleukin immunotherapy agent. This includes methods for treating or relieving cancer symptoms in human or animal subjects in need.

[0066] In an additional aspect, the present invention includes the use of compositions comprising agents for inhibiting or suppressing TGF-β2 expression in combination with checkpoint inhibitors and interleukin immunotherapies in the preparation of pharmaceuticals for treating or relieving cancer symptoms in human subjects or animals.

[0067] In additional cases, agents that inhibit or suppress TGF-β2 expression, immune checkpoint inhibitors, and interleukin immunotherapies can be administered together, simultaneously, sequentially, or at different times.

[0068] Human TGF-β2-specific phosphorothioate antisense oligodeoxynucleotide Antisense oligonucleotides (ASOs) can be single-stranded deoxyribonucleotides and may be complementary to mRNA targets. Antisense therapy can downregulate molecular targets, which can be achieved by inducing RNase H endonuclease activity that cleaves RNA-DNA heteroduplexes and significantly reduces translation of target genes. Other ASO mechanisms include inhibition of 5' cap formation, alteration of splicing processes such as splice switching, and steric hindrance of ribosome activity.

[0069] Antisense therapeutic strategies can utilize single-stranded DNA oligonucleotides that inhibit protein synthesis by mediating catalytic degradation of target mRNA or by binding to sites on mRNA required for translation. Antisense oligonucleotides can be designed to target viral RNA genomes or viral transcripts. Antisense oligonucleotides can provide an approach for identifying potential targets and therefore may constitute potential therapeutic agents.

[0070] Antisense oligonucleotides can be small synthetic fragments of single-stranded DNA, possibly 15–30 nucleotides in length. ASOs can specifically bind to complementary DNA / RNA sequences via Watson-Crick hybridization, and once bound to the target RNA, they inhibit the translation process by either inducing cleavage mechanisms or inhibiting mRNA maturation. ASOs can selectively inhibit gene expression with specificity. Chemical modifications to the DNA or RNA can be used to increase stability.

[0071] For example, modifications can be introduced into phosphodiester bonds, sugar rings, and the backbone. ASO antivirals can block the translation process by either (i) cleavage of mRNA mediated by ribonuclease H (RNAse H) or RNase P, or (ii) steric (non-binding) blockage of enzymes involved in the translation of the target gene. Human TGF-β2-specific phosphorothioate antisense oligodeoxynucleotide (OT-101; AP 12009; Travedersen), hereinafter referred to as OT-101 or AP 12009, aims to reduce the level of TGF-β2 protein in malignant gliomas, thereby slowing disease progression.

[0072] Antisense oligodeoxynucleotides are short strings of DNA designed to downregulate gene expression by interfering with the translation of specific encoded proteins at the mRNA level. OT-101 is a synthetic 18-mer phosphorothioate oligodeoxynucleotide (S-ODN) in which all 3'-5' links are modified to phosphorothioate. The molecular formula is C 177 H 208 N 60 Na 17 O 94 P 17 S 17 Its molecular weight is 6,143 g / mol. OT-101 can be designed to be complementary to a specific sequence of human TGF-β2 mRNA after gene expression.

[0073] OT-101 can be supplied as a lyophilized powder in 50 mL glass vials in three different quantities. Each vial is identified by the clinical trial drug name, trial number, dosing group, application method, OT-101 content (mg), total volume after dissolution (mL) and the resulting concentration (μM), sponsor name, manufacturer name, batch number, vial number, storage temperature, and expiration date. The investigational drug can be supplied in sealed units, separately packaged for each concentration. The packaging may contain the appropriate vial and all necessary components of the application system (i.e., syringe, tubing, and filter). Before use, the lyophilized OT-101 powder may be dissolved in an isotonic (0.9%) aqueous sodium chloride solution.

[0074] Examples of agents of this disclosure for inhibiting or suppressing TGF-β2 expression include TGF-β2-specific antisense oligonucleotides shown in Table 1, SEQ ID NO: 1-136.

[0075] (Table 1) TGF-β2-specific antisense oligonucleotides TIFF2026519788000003.tif101147TIFF2026519788000004.tif235147TIFF2026519788000005.tif235147TIFF2026519788000006.tif235147

[0076] As is well known in the art, the sequences in Table 1 can be chemically modified to provide their active variants, their LNA variants, and their gap mar variants. The sequences in Table 1 can be used as active agents in any combination, for example, a pooled combination.

[0077] It will be understood that additional antisense oligonucleotides can be constructed based on the TGF-β2 gene sequence in this disclosure.

[0078] In some embodiments, the antisense sequence agent may be a gapmer formed by adding 1 to 5 protected ribonucleotides to each adjacent region of the phosphorothioate deoxynucleotide sequence in Table 1. For example, the ribonucleotides can be protected with 2'-OMe, 2'-OEt, or 2'-O-MOE substituents, or with LNA, cMOE, or cEt bridges, as well as phosphorothioate linkages.

[0079] In some embodiments, the antisense sequence agent may be an nMn RNA(2'-OMe)*-DNA*-RNA(2'-OMe)* gapmer, where n is 3 to 7 and M is 6 to 12. In certain embodiments, the gapmer may be a 3-10-3 or 5-10-5 LNA*-DNA*-LNA* or cEt*-DNA*-cEt* gapmer (* represents phosphorothioate linkage).

[0080] Examples of agents of this disclosure for inhibiting or suppressing TGF-β2 expression include TGF-β2-specific phosphorothioate antisense oligonucleotides, shown as SEQ ID NO: 137-144 in Table 2, based on the sequences in Table 1.

[0081] (Table 2) TGF-β2 specific phosphorothioate antisense oligonucleotides TIFF2026519788000007.tif55170

[0082] Aspects of the present invention further include pharmaceutical compositions for inhibiting or suppressing TGF-β expression in humans or animals, or for treating or relieving cancer symptoms. The pharmaceutical compositions may contain a TGF-β inhibitor, artemisinin, a pharmaceutically acceptable salt form, ester, polymorph, or stereoisomer thereof, and any combination thereof, as well as a carrier. The TGF-β inhibitor may be selected from TGF-β2-specific antisense oligonucleotides. The carrier may be sterile water for injection, saline, isotonic saline, or a combination thereof.

[0083] Importantly, the compositions of this disclosure may be substantially free of excipients. Compositions of the present invention that are substantially free of excipients have been found to be remarkably stable in a carrier. In some embodiments, the compositions may be stable at 37°C in a carrier for at least 14 days, or at least 21 days, or at least 28 days.

[0084] In additional embodiments, the pharmaceutical composition for injection may contain less than 1% by weight of an excipient, or less than 0.5% by weight of an excipient, or less than 0.1% by weight of an excipient.

[0085] OT-101 Antisense Oligonucleotide API Travedersen / OT-101 is a synthetic 18-mer S-ODN consisting of the bases adenine (A), thymine (T), guanine (G), and cytosine (C), with all 3'-5' links modified to phosphorothioates. Thioate modification can make the drug more resistant to degradation, resulting in increased stability in vitro and in vivo. Its molecular structure (nucleotide sequence) can be designed to be complementary to a specific sequence of human transforming growth factor-beta-2 (TGF-β2) mRNA. To achieve the best antisense effect in vitro and in vivo, this sequence can be selected from relevant molecules based on its superior chemical and structural properties, bioactivity, and specificity.

[0086] Table 3 shows the chemical structure, exemplary phosphorothioate moiety (CAG), and physical characteristics of trabedersene.

[0087] (Table 3) Chemical and physical characteristics of Travedersene TIFF2026519788000008.tif106162

[0088] IMPs may be supplied as sterile lyophilized products for injection solutions in 50H glass vials (primary containers) containing 7.37 mg of travedersen (intratumoral treatment) and in 20R glass vials (primary containers) containing 250 mg of travedersen (intravenous treatment), respectively. Excipients may be absent from the finished drug product. Glass vials are commonly used for parenteral administration. Glass vials can be sealed with sterile rubber stoppers suitable for lyophilization. Stoppers may be sealed with crimping caps, including colored flip-off caps. For clinical use, each vial may be provided in a white collapsible box to protect it from light exposure and damage during transport. Both glass vials and collapsible boxes can be labeled according to local requirements. Primary and secondary containers of the closure system may meet international quality standards for packaging of sterile solid drug products for injection.

[0089] Immune checkpoint inhibitors As referred to herein, checkpoint inhibitors, which are known in the art, are immune checkpoint inhibitors. Checkpoint inhibitors are immunotherapeutic drugs that block checkpoint proteins from binding to their partner proteins. This prevents the "off" signal from being sent, thereby allowing T cells to kill cancer cells. More specifically, checkpoint proteins such as PD-1 on T cells suppress the immune response. The binding of PD-L1 to PD-1 prevents T cells from killing tumor cells. Therefore, by blocking the binding of PD-L1 to PD-1 with immune checkpoint inhibitors, T cells may be able to kill tumor cells. The immune system can essentially be reactivated, and as a result, T cells can attack cancer cells.

[0090] In some embodiments, the checkpoint inhibitors of the present disclosure may be inhibitors of PD-1, CTLA-4, or PD-L1.

[0091] In certain embodiments, the checkpoint inhibitors of this disclosure may be inhibitors of PD-1.

[0092] In certain embodiments, the checkpoint inhibitor of this disclosure may be pembrolizumab.

[0093] In certain embodiments, the checkpoint inhibitors of this disclosure may be pembrolizumab, nivolumab, semiprimab, spartalizumab, atezolizumab, avelumab, or durvalumab.

[0094] While we do not wish to be constrained by theory, PD-1 receptor-ligand interactions could be a major pathway for tumor hijacking and suppression of immune regulation. Under healthy conditions, the normal function of PD-1 expressed on the surface of activated T cells is to downmodulate unwanted or excessive immune responses, including autoimmune reactions. Following T cell stimulation, PD-1 recruits the tyrosine phosphatases SHP-1 and SHP-2 to an immune receptor tyrosine-based switch motif in its cytoplasmic tail, which results in the dephosphorylation of effector molecules involved in the CD3 T cell signaling cascade, such as CD3 zeta (CD3ζ), protein kinase C-theta (PKCθ), and zeta chain-associated protein kinase (ZAP70).

[0095] Active IL-2 immunotherapy agents In certain embodiments, the interleukin immunotherapy agents of this disclosure may be natural or synthetic IL-2, high-dose IL-2, recombinant IL-2, or aldesleukin.

[0096] In certain embodiments, the interleukin immunotherapy agent of this disclosure may be recombinant human IL-2 protein.

[0097] In an additional embodiment, the interleukin immunotherapy agents of the present disclosure may be IL-2 proteins that have been modified to reduce toxicity or off-target effects, or to have a target-directed moiety attached, or to be engineered to target specific cells.

[0098] Numbered embodiments of the present invention include: (1) A method for treating or relieving the symptoms of cancer in a person in need, A step of administering a composition containing an agent for inhibiting or suppressing the expression of TGF-β2; The process of administering an immune checkpoint inhibitor; and The process of administering interleukin immunotherapy agents. Methods that include... (2) Agents for inhibiting or suppressing TGF-β2 expression, in combination with immune checkpoint inhibitors and active interleukin immunotherapies, for use in treating or relieving the symptoms of cancer. (3) A composition comprising an agent for inhibiting or suppressing the expression of TGF-β2 for use in the preparation of a pharmaceutical product or for treating or relieving cancer symptoms in a subject, in combination with an immune checkpoint inhibitor and an active interleukin immunotherapy agent, and a pharmaceutically acceptable carrier. (4) A method, agent, or composition according to any of embodiments 1 to 3, wherein the cancer is a solid tumor, pancreatic cancer, melanoma, lung cancer, breast cancer, multiple myeloma, or colorectal cancer. (5) A method, agent, or composition according to any of embodiments 1 to 4, wherein the agent for inhibiting or suppressing the expression of TGF-β2, the immune checkpoint inhibitor, and the active interleukin immunotherapy agent are administered together, simultaneously, sequentially, or at separate times. (6) A method, agent, or composition according to any of embodiments 1 to 5, wherein the composition and agent are administered by infusion or injection. (7) A method, agent, or composition according to any of embodiments 1 to 6, wherein the agent for inhibiting or suppressing the expression of TGF-β2 is selected from Table 1 or Table 2, and their chemically modified variants, their LNA variants, their gap mar variants, and any combination or pool thereof. (8) The agent for inhibiting or suppressing the expression of TGF-β2, A method, agent, or composition according to any of embodiments 1 to 7, which is TIFF2026519788000009.tif12128. (9) A method, agent, or composition according to any of embodiments 1 to 8, wherein the agent or composition comprises a carrier which is sterile water for injection, physiological saline, isotonic physiological saline, or a combination thereof. (10) A method, agent, or composition according to any of embodiments 1 to 9, wherein the agent or composition is substantially free of excipients. (11) A method, agent, or composition according to any of embodiments 1 to 10, wherein the composition is stable in a carrier at 37°C for at least 14 days. (12) A method, agent, or composition according to any of embodiments 1 to 11, wherein the immune checkpoint inhibitor is an inhibitor of PD-1, CTLA-4, or PD-L1. (13) A method, agent, or composition according to any of embodiments 1 to 12, wherein the immune checkpoint inhibitor is pembrolizumab, nivolumab, semiprimab, spartalizumab, atezolizumab, avelumab, or durvalumab. (14) A method, agent, or composition according to any of embodiments 1 to 13, wherein the active interleukin immunotherapy agent is natural IL-2, high-dose IL-2, recombinant IL-2, or aldesleukin. (15) A method, agent, or composition according to any of embodiments 1 to 14, wherein, when administered or used, the subject has reduced TGF-β2 expression. (16) Selecting subjects who will benefit from the method, agent, or use using one or more biomarkers. A method, agent, or composition according to any of embodiments 1 to 15, including the above. (17) A method, agent, or composition according to any of embodiments 1 to 16, wherein the one or more biomarkers are IRF5 levels, ITGAM levels, or a combination thereof. (18) The one or more biomarkers are ITGAM, and The subject is selected when the expression of ITGAM is at a level exceeding that seen in healthy patients. A method, agent, or composition according to any of embodiments 1 to 17. (19) A method, agent, or composition according to any of embodiments 1 to 18, wherein, after administration or use, the subject has an increased level of IRF5. (20) A method, agent, or composition according to any of embodiments 1 to 19, wherein, after the administration or use, the subject has an increased level of ITGAM. (21) Administering to the subject a therapeutically sufficient amount of a pharmaceutical composition containing an expression product that is IRF5 or ITGAM. A method, agent, or composition according to any of embodiments 1 to 20, including the above. (22) A method, agent, or composition according to any of embodiments 1 to 21, wherein the expression product is mRNA, polypeptide, protein, or a fragment thereof, or a combination thereof. (23) A method, agent, or composition according to any of embodiments 1 to 22, wherein the administration or use reduces mortality at 6, 12, 18, 24, 30, or 36 months. (24) A method, agent, or composition according to any of embodiments 1 to 23, wherein the administration or use increases the survival rate at 6, 12, 18, 24, 30, or 36 months. (25) A method, agent, or composition according to any of embodiments 1 to 24, wherein the administration or use of the composition is combined with a standard treatment for cancer, the standard treatment comprising chemotherapy or radiotherapy.

[0099] All publications, including patents, patent application publications, and non-patent publications, as well as sequence listings, referred to herein are expressly incorporated herein by reference in their entirety for any purpose.

[0100] While the aforementioned disclosures have been described in detail as examples for the purpose of clarifying understanding, it will be apparent to those skilled in the art that certain changes and modifications are included in this disclosure and that they can be implemented within the scope of the appended claims, which are presented as examples rather than limitations, without excessive experimentation. The present invention includes all such additional embodiments, equivalents, and modifications. The present invention includes any combination or mixture of various exemplary components, examples, and features, materials, elements, or limitations in the claimed embodiments.

[0101] The designations of agents, compositions, and structures in this disclosure encompass all possible isomers, stereoisomers, diastereomers, enantiomers, and / or optical isomers understood to exist for any given structure, including racemics or any other mixtures thereof. [Examples]

[0102] Example 1. This example demonstrates that ITGAM is a biomarker for melanoma patients who would benefit from a therapeutic combination of TGF-β2-specific antisense agents, IL-2 immunotherapy agents, and PD-1 checkpoint inhibitors.

[0103] Figure 1 shows the use of ITGAM as a biomarker for melanoma patients who benefit from a therapeutic combination of TGF-β2-specific antisense agents, IL-2 immunotherapy agents, and PD-1 checkpoint inhibitors.

[0104] Figure 1 shows the Kaplan-Meier overall survival chart (KM plotter) obtained in a study of clinical outcomes in melanoma patients. Figure 1 shows that the use of PD-1 checkpoint inhibitors in selected patients with high ITGAM was associated with improved survival in the case of low TGF-β2 (log-rank P=0.0039).

[0105] Example 2. This example demonstrates that ITGAM is a biomarker for pancreatic cancer patients who would benefit from a therapeutic combination of TGF-β2-specific antisense agents, IL-2 immunotherapy agents, and PD-1 checkpoint inhibitors.

[0106] Figure 2 shows the use of ITGAM as a biomarker for pancreatic cancer patients who benefit from a therapeutic combination of TGF-β2-specific antisense agents, IL-2 immunotherapy agents, and PD-1 checkpoint inhibitors.

[0107] Figure 2 shows the Kaplan-Meier overall survival chart (KM plotter) obtained in a study of clinical outcomes in pancreatic cancer. Figure 2 shows that improved survival was demonstrated in the case of high IL-2 use in selected patients with high ITGAM (log-rank P=0.034).

[0108] Example 3. This example demonstrates that IRF5 is a biomarker for melanoma patients who benefit from a therapeutic combination of TGF-β2-specific antisense agents, IL-2 immunotherapy agents, and PD-1 checkpoint inhibitors.

[0109] Figure 3 shows the use of IRF5 as a biomarker for melanoma patients who benefit from a therapeutic combination of TGF-β2-specific antisense agents, IL-2 immunotherapy agents, and PD-1 checkpoint inhibitors.

[0110] Figure 3 shows the Kaplan-Meier overall survival chart obtained in a study of clinical outcomes for melanoma (KM plotter). Figure 3 shows that the use of PD-1 checkpoint inhibitors in selected patients with high IRF5 was associated with improved survival in the case of low TGF-β2 (log-rank P = 0.00053).

[0111] Example 4. This example demonstrates that a therapeutic combination of a TGF-β2-specific antisense agent, an IL-2 immunotherapy agent, and a PD-1 checkpoint inhibitor improves survival in cancer.

[0112] Figure 4 shows the impact of therapeutic combinations of TGF-β2-specific antisense agents, IL-2 immunotherapies, and PD-1 checkpoint inhibitors on overall survival in cancer.

[0113] Figure 4 shows the Kaplan-Meier overall survival chart (KM plotter) obtained from a study of clinical outcomes in cancer patients. This study included several types of tumors: bladder (N=73), esophageal adenocarcinoma (N=103), glioblastoma (N=28), hepatocellular carcinoma (N=22), HNSCC (N=5), melanoma (N=423), NSCLC (N=21), NSCL (N=22), and urothelial (N=348). Figure 4 shows that improved survival was observed with respect to the use of PD-1 checkpoint inhibitors when the ratio of IL-2 to TGF-β2 (IL-2 / TGF-β2) was high (log-rank P=0.0031).

[0114] Referring to Figure 4, the median overall survival for the high-expression cohort, i.e., high IL-2 / TGF-β2, increased dramatically to 23 months compared to 10 months for the low-expression cohort, i.e., low IL-2 / TGF-β2 cohort.

[0115] This study demonstrated an unexpectedly favorable impact on overall survival and established a basis for therapeutic combinations of TGF-β2-specific antisense agents, IL-2 immunotherapies, and PD-1 checkpoint inhibitors in cancer.

[0116] Example 5. This example demonstrates that a therapeutic combination of a TGF-β2-specific antisense agent, an IL-2 immunotherapy agent, and a PD-1 checkpoint inhibitor improves survival in melanoma.

[0117] Figure 5 shows the impact of therapeutic combinations of TGF-β2-specific antisense agents, IL-2 immunotherapies, and PD-1 checkpoint inhibitors on overall survival in melanoma.

[0118] Figure 5 shows the Kaplan-Meier overall survival chart obtained in a study of the clinical outcomes of melanoma (KM plotter). Figure 5 shows that improved survival was observed when the ratio of IL-2 to TGF-β2 (IL-2 / TGF-β2) was high with the use of PD-1 checkpoint inhibitors (log-rank P = 3.7e-06).

[0119] Referring to Figure 5, the median overall survival for the high-expression cohort, i.e., high IL-2 / TGF-β2, increased dramatically from 20 months to 31 months, compared to 20 months for the low-expression cohort, i.e., low IL-2 / TGF-β2 cohort.

[0120] This study demonstrated an unexpectedly favorable impact on overall survival and established a basis for therapeutic combinations of TGF-β2-specific antisense agents, IL-2 immunotherapies, and PD-1 checkpoint inhibitors in melanoma.

[0121] Example 6. This example demonstrates that a therapeutic combination of a TGF-β2-specific antisense agent, an IL-2 immunotherapy agent, and a PD-1 checkpoint inhibitor improves survival in melanoma.

[0122] Figure 6 shows the Kaplan-Meier overall survival chart (KM plotter) obtained from a study of clinical outcomes in melanoma patients. Figure 6 shows that improved survival was observed in cases of high IL-2 with the use of immune checkpoint inhibitors (log-rank P=0.0015).

[0123] Example 7. This example demonstrates that a therapeutic combination of a TGF-β2-specific antisense agent, an IL-2 immunotherapy agent, and a PD-1 checkpoint inhibitor improves survival in melanoma.

[0124] Figure 7 shows the Kaplan-Meier overall survival chart (KM plotter) obtained from a study of clinical outcomes in melanoma patients. Figure 7 shows that improved survival was observed in patients with low TGF-β2 levels with the use of immune checkpoint inhibitors (log-rank P=0.006).

[0125] Example 8. This example demonstrates that a therapeutic combination of a TGF-β2-specific antisense agent, an IL-2 immunotherapy agent, and a PD-1 checkpoint inhibitor improves survival in cancer.

[0126] Figure 8 shows the Kaplan-Meier overall survival chart obtained in a study of clinical outcomes in cancer patients (KM plotter). Figure 8 shows that improved survival was observed in cases of high IL-2 with the use of PD-1 checkpoint inhibitors (log-rank P=0.007).

[0127] Example 9. This example demonstrates that a therapeutic combination of a TGF-β2-specific antisense agent, an IL-2 immunotherapy agent, and a PD-1 checkpoint inhibitor improves survival in cancer.

[0128] Figure 9 shows the Kaplan-Meier overall survival chart obtained in a study of clinical outcomes in cancer patients (KM plotter). Figure 9 shows that improved survival was demonstrated in the case of low TGF-β2 with the use of PD-1 checkpoint inhibitors (log-rank P=0.0028).

[0129] Example 10. This example demonstrates that a therapeutic combination of a TGF-β2-specific antisense agent, an IL-2 immunotherapy agent, and a PD-1 checkpoint inhibitor improves survival in cancer.

[0130] Figure 10 shows the Kaplan-Meier overall survival chart (KM plotter) obtained from a study of clinical outcomes in melanoma patients. This study included multiple tumor types. Figure 10 shows that improved survival was observed with respect to the use of immune checkpoint inhibitors when the ratio of IL-2 to TGF-β2 (IL-2 / TGF-β2) was high (log-rank P = 3.7e-06). Therefore, therapeutic combinations of TGF-β2-specific antisense agents, IL-2 immunotherapies, and PD-1 checkpoint inhibitors improve survival in cancer.

[0131] Example 11. This example demonstrates that a therapeutic combination of a TGF-β2-specific antisense agent, an IL-2 immunotherapy agent, and a PD-1 checkpoint inhibitor improves survival in cancer.

[0132] Figure 11 shows the Kaplan-Meier overall survival chart obtained from a study of clinical outcomes in cancer patients (KM plotter). Figure 11 shows that improved survival was observed when the ratio of IL-2 to TGF-β2 (IL-2 / TGF-β2) was high with the use of PD-1 checkpoint inhibitors (log-rank P = 2.2e-08). Therefore, therapeutic combinations of TGF-β2-specific antisense agents, IL-2 immunotherapies, and PD-1 checkpoint inhibitors improve survival in cancer.

Claims

1. A method for treating or relieving the symptoms of cancer in a subject in need, A step of administering a composition containing an agent for inhibiting or suppressing the expression of TGF-β2; The process of administering an immune checkpoint inhibitor; and The process of administering interleukin immunotherapy agents. Methods that include...

2. Agents for inhibiting or suppressing TGF-β2 expression, in combination with immune checkpoint inhibitors and active interleukin immunotherapies, for use in treating or relieving cancer symptoms.

3. A composition comprising an agent for inhibiting or suppressing TGF-β2 expression for use in the preparation of pharmaceuticals, or for treating or relieving cancer symptoms in a subject, in combination with an immune checkpoint inhibitor and an active interleukin immunotherapy agent, and a pharmaceutically acceptable carrier.

4. A method, agent, or composition according to any one of claims 1 to 3, wherein the cancer is a solid tumor, pancreatic cancer, melanoma, lung cancer, breast cancer, multiple myeloma, or colorectal cancer.

5. A method, agent, or composition according to any one of claims 1 to 3, wherein the agent for inhibiting or suppressing the expression of TGF-β2, the immune checkpoint inhibitor, and the active interleukin immunotherapy agent are administered together, simultaneously, sequentially, or at separate times.

6. A method, agent, or composition according to any one of claims 1 to 3, wherein the composition and agent are administered by infusion or injection.

7. A method, agent, or composition according to any one of claims 1 to 3, wherein the agent for inhibiting or suppressing the expression of TGF-β2 is selected from Table 1 or Table 2, and their chemically modified variants, their LNA variants, their gap mar variants, and any combination or pool thereof.

8. The agent for inhibiting or suppressing the expression of TGF-β2 is A method, agent, or composition according to any one of claims 1 to 3.

9. A method, agent, or composition according to any one of claims 1 to 3, wherein the agent or composition comprises a carrier which is sterile water for injection, physiological saline, isotonic physiological saline, or a combination thereof.

10. A method, agent, or composition according to any one of claims 1 to 3, wherein the agent or composition is substantially free of excipients.

11. A method, agent, or composition according to any one of claims 1 to 3, wherein the composition is stable in a carrier at 37°C for at least 14 days.

12. A method, agent, or composition according to any one of claims 1 to 3, wherein the immune checkpoint inhibitor is an inhibitor of PD-1, CTLA-4, or PD-L1.

13. A method, agent, or composition according to any one of claims 1 to 3, wherein the immune checkpoint inhibitor is pembrolizumab, nivolumab, semiprimab, spartalizumab, atezolizumab, avelumab, or durvalumab.

14. A method, agent, or composition according to any one of claims 1 to 3, wherein the active interleukin immunotherapy agent is natural IL-2, high-dose IL-2, recombinant IL-2, or aldesleukin.

15. A method, agent, or composition according to any one of claims 1 to 3, wherein, upon administration or use, the subject has reduced TGF-β2 expression.

16. Selecting subjects who will benefit from the method, agent, or use using one or more biomarkers. A method, agent, or composition according to any one of claims 1 to 3, comprising:

17. The method, agent, or composition according to claim 16, wherein the one or more biomarkers are IRF5 levels, ITGAM levels, or a combination thereof.

18. The one or more of the aforementioned biomarkers are ITGAM, and The subject is selected when the expression of ITGAM is at a level exceeding that seen in healthy patients. The method, agent, or composition according to claim 16.

19. A method, agent, or composition according to any one of claims 1 to 3, wherein, after the administration or use, the subject has an increased level of IRF5.

20. A method, agent, or composition according to any one of claims 1 to 3, wherein, after the administration or use, the subject has an increased level of ITGAM.

21. Administering to the subject a therapeutically sufficient amount of a pharmaceutical composition containing an expression product that is IRF5 or ITGAM. A method, agent, or composition according to any one of claims 1 to 3, comprising:

22. The method, agent, or composition according to claim 21, wherein the expression product is mRNA, polypeptide, protein, or a fragment thereof, or a combination thereof.

23. A method, agent, or composition according to any one of claims 1 to 3, wherein the administration or use reduces mortality at 6, 12, 18, 24, 30, or 36 months.

24. A method, agent, or composition according to any one of claims 1 to 3, wherein the administration or use increases the survival rate at 6, 12, 18, 24, 30, or 36 months.

25. The method, agent, or composition according to any one of claims 1 to 3, wherein the administration or use of the composition is combined with a standard treatment for cancer, the standard treatment comprising chemotherapy or radiotherapy.

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