Cancer therapy with TGF-β-2 and irinotecan

Abstract

The present invention describes a method for treating or improving the symptoms of cancer in subjects with agents, compositions, and regimens designed to promote anti-tumor effects and avoid drug resistance.A combination of active agents can be used, including agents, regimens, or preparations containing irinotecan, such as FOLFIRINOX, NALIRIFOX, IRIFOX, or IRINOX, and agents for inhibiting or suppressing the expression of TGF-β2.Biomarkers can be used to select subjects who will benefit from such therapeutic agents. TIFF2026507184000026.tif117156

Description

[Technical Field]

[0001] Sequence Listing This application contains a Sequence Listing that was filed electronically as an ST.26 file with file name 018988-008WO1_SL.xml and is 120,202 bytes in size, created on February 21, 2024.

[0002] Technical Field The present invention describes a method for treating or ameliorating the symptoms of cancer in human or animal subjects by agents and compositions designed to promote anti-tumor activity.Exemplary synergistic pharmaceutical treatments include a combination of active agents, including an agent for inhibiting or suppressing the expression of TGF-β2, combined with an agent or formulation containing irinotecan.Biomarkers can be used to select subjects who will benefit from the agent, use, or method. [Background technology]

[0003] background Cancer is a complex pathology that involves multiple different cellular pathways. Due to this complexity, it has been difficult to find an effective therapy that can have sufficient anti-tumor activity against various cancers. What's worse, cancer cells and malignant tumors may develop resistance to various agents and drugs, which may cause patients to become unresponsive.

[0004] Irinotecan is an example of a conventional therapy that has been used for various malignant tumors. The drawbacks of conventional therapies, such as compositions containing irinotecan, include a lack of sufficient efficacy against a wide range of cancers. Additional drawbacks include the development of resistance to irinotecan, which can limit successful treatment outcomes. An additional drawback of using compositions containing irinotecan is cross-resistance to other anticancer drugs. Furthermore, not all cancer patients reliably respond to irinotecan-based therapy and must instead receive palliative care. These drawbacks can limit available treatment options.

[0005] What is needed to overcome these difficulties are methods and uses that employ combinations of compositions and agents that enhance efficacy against malignancies and avoid the effects of resistance.

[0006] There is an urgent need for compositions, agents, uses and methods that combine various agents with significant anti-tumor activity.There is also a need for improved guidance in the use of such compositions and agents by using appropriate clinically validated biomarkers to provide synergistic improvements in efficacy. Summary of the Invention

[0007] Quick Overview The present invention describes methods for treating or ameliorating the symptoms of cancer in human or animal subjects with agents and compositions designed to promote anti-tumor activity and circumvent drug resistance effects against a variety of different cancers.

[0008] Embodiments of the present disclosure include agents and methods for treating or ameliorating symptoms of cancer in a human or animal subject in need thereof with agents for inhibiting or suppressing the expression of TGF-β2.

[0009] The synergistic pharmaceutical treatment methods of the present invention include combinations of active agents, such as agents for inhibiting or suppressing the expression of TGF-β2, and agents such as irinotecan and formulations containing irinotecan.

[0010] In some embodiments, the biomarkers can be used to select subjects who will benefit from an agent, use, or method.

[0011] In further embodiments, the agents and compositions can be used in combination with chemotherapy, radiation therapy, and other standard of care therapies.

[0012] The present disclosure includes the use of a composition containing an agent for inhibiting or suppressing the expression of TGF-β2 in the preparation of a medicament for treating or ameliorating symptoms of cancer in a human or animal subject.

[0013] The present invention contemplates agents, uses, and methods for treating or ameliorating the symptoms of a variety of different cancers, including pancreatic cancer, colorectal cancer, gastric cancer, cervical cancer, lung cancer, melanoma, skin cancer, breast cancer, prostate cancer, kidney cancer, ovarian cancer, thymic cancer, and multiple myeloma.

[0014] Aspects of the present invention include the following.

[0015] An antisense agent for inhibiting or suppressing the expression of TGF-β2, in combination with an agent comprising irinotecan, for use in treating or ameliorating symptoms of cancer in a subject.

[0016] Use of an antisense agent for inhibiting or suppressing the expression of TGF-β2 in combination with an agent comprising irinotecan in the preparation of a medicament for treating or ameliorating symptoms of cancer in a subject.

[0017] 1. A method for treating or ameliorating a symptom of cancer in a subject in need thereof, comprising: administering to the subject a therapeutically effective amount of an antisense agent to inhibit or suppress expression of TGF-β2; and administering to said subject a therapeutically effective amount of an agent comprising irinotecan. A method comprising:

[0018] The agent, use, or method described above, wherein the agent containing irinotecan is a FOLFIRINOX agent, a NALIRIFOX agent, an IRIFOX agent, or an IRINOX agent.

[0019] The agent, use, or method as described above, wherein the agent comprising irinotecan comprises a nanosized particle, a nanosized micelle, a nanosized liposome, or a nanosized exosome.

[0020] The agent, use or method as described above, wherein the agent for inhibiting or suppressing the expression of TGF-β2 and the agent comprising irinotecan are administered contemporaneously, simultaneously, sequentially or at separate times.

[0021] The agent, use, or method as described above, wherein the agent for inhibiting or suppressing the expression of TGF-β2 and the agent comprising irinotecan are administered separately or in combination by injection or infusion.

[0022] The agent, use or method as described above, wherein the cancer is pancreatic cancer, colorectal cancer, gastric cancer, cervical cancer, lung cancer, melanoma, skin cancer, breast cancer, prostate cancer, kidney cancer, ovarian cancer, thymic cancer, and multiple myeloma.

[0023] The agent, use, or method described above, wherein the agent for inhibiting or suppressing the expression of TGF-β2 is a TGF-β2-specific antisense oligonucleotide that is complementary to the TGF-β2 transcript and is 15 to 30 nucleotides in length.

[0024] The agent, use, or method described above, wherein the agent for inhibiting or suppressing the expression of TGF-β2 is a TGF-β2-specific antisense oligonucleotide that is complementary to TGF-β2 pre-RNA, pre-mRNA, or mRNA and is 18 to 21 nucleotides in length.

[0025] The agent, use, or method as described above, wherein the agent for inhibiting or suppressing expression of TGF-β2 is one or more TGF-β2-specific antisense oligonucleotides complementary to a TGF-β2 transcript as in Table 1, and chemically modified variants thereof, LNA variants thereof, gapmer variants thereof, and any combination or pool thereof.

[0026] The agent, use or method as described above, wherein the TGF-β2 specific antisense oligonucleotide has no more than one or no more than two mismatches compared to the target human TGF-β2.

[0027] The agent, use or method as described above, wherein the TGF-β2 specific antisense oligonucleotide reduces the level of TGF-β2 transcript by at least 60%, or at least 70%, or at least 80%, or at least 90%.

[0028] The agent, use, or method described above, wherein the TGF-β2-specific antisense oligonucleotide reduces the level of any TGF-β1 transcript and the level of any TGF-β3 transcript by less than 10%, or less than 5%, or less than 1%.

[0029] TGF-β2 specific antisense oligonucleotides One or more nucleotides chemically modified as a phosphorothioate internucleoside linkage, a methoxypropylphosphonate internucleoside linkage, an aminophosphoro linkage to a morpholino group, a 2'-OMe ribose group, a 2'-MOE methoxyethyl ribose group, a 2'-4' constrained methoxyethyl bicyclic ribose group, a 2'-4' constrained ethyl bicyclic ribose group, an LNA ribose group, a 2'-F ribose group, or a 5-methylcytosine base The agent, use or method as described above, comprising:

[0030] An agent, use or method as described above, wherein the antisense agent is conjugated to polyethylene glycol, a lipid, or triantennary N-acetyl-galactosamine.

[0031] The agent, use, or method described above, wherein the agent for inhibiting or suppressing the expression of TGF-β2 comprises a carrier that is sterile water for injection, normal saline, isotonic saline, or a combination thereof, which may be the same or different for each agent.

[0032] The agent, use or method as described above, wherein the agent for inhibiting or suppressing the expression of TGF-β2 is substantially excipient-free.

[0033] The agent, use or method as described above, wherein the agent for inhibiting or suppressing the expression of TGF-β2 is stable in a substantially excipient-free carrier at 37°C for at least 14 days.

[0034] The agent, use or method as described above, wherein the patient is selected to have been previously treated with any agent including irinotecan.

[0035] An agent, use or method as described above, comprising using one or more biomarkers to select subjects who will benefit from the agent, use or method.

[0036] An agent, use or method as described above, wherein patients having elevated levels of TGF-β2 are selected.

[0037] The agent, use, or method as described above, wherein the one or more biomarkers are levels of tumor mutational burden (TMB), levels of tumor neoantigens, levels of clinical hypoxia, or a combination thereof.

[0038] The agent, use, or method as described above, wherein the one or more biomarkers are the level of tumor gene mutation burden, the level of reduced neoantigen load measured in the tumor microenvironment, the level of reduced macrophages measured in the tumor microenvironment, the level of reduced mesenchymal stem cells (MSCs) measured in the tumor microenvironment, the level of type 2 helper T cells (Th2) measured in the tumor microenvironment, and combinations thereof.

[0039] The above-mentioned method, agent, or use, wherein a subject receiving the administration or use has an improved level of at least one of the one or more biomarkers compared to the level found in a healthy patient.

[0040] An agent, use or method as described above, wherein the administration or use reduces mortality at 6, 12, 18, 24, 30 or 36 months.

[0041] An agent, use or method as described above, wherein the administration or use increases survival at 6, 12, 18, 24, 30 or 36 months.

[0042] An agent, use or method as described above, wherein administration or use improves health-related quality of life (HRQoL) by 15%, or 25%, or 35%, or 45%.

[0043] The above-mentioned agents, uses or methods in combination with any one or more pharmaceutical agents including cancer-targeting agents, cancer growth blockers, EGFR inhibitors, and combinations thereof.

[0044] The above-mentioned agent, use, or method in combination with any one or more medicaments selected from bevacizumab, everolimus, velzutifan, dabrafenib, trametinib, and combinations thereof.

[0045] The above-mentioned agent, use, or method in combination with any one or more medicaments that are cancer growth blockers selected from angiogenesis inhibitors, histone deacetylase inhibitors, hedgehog blockers, mTOR inhibitors, p53 inhibitors, PARP inhibitors, proteasome inhibitors, tyrosine kinase inhibitors, and combinations thereof.

[0046] The above-mentioned agent, use, or method in combination with any one or more medicaments which are EGFR inhibitors selected from erlotinib, gefitinib, afatinib, osimertinib, dacomitinib, and combinations thereof.

[0047] The agent, use or method as described above in combination with a standard of care treatment for cancer.

[0048] The above mentioned agent, use or method in combination with radiation therapy or electric field therapy.

[0049] a therapeutically effective amount of an antisense agent for inhibiting or suppressing the expression of TGF-β2, and Therapeutically effective amount of an agent comprising irinotecan A kit for treating or ameliorating a symptom of cancer, comprising:

[0050] The kit as described above, wherein the cancer is pancreatic cancer, colorectal cancer, gastric cancer, cervical cancer, lung cancer, melanoma, skin cancer, breast cancer, prostate cancer, kidney cancer, ovarian cancer, thymic cancer, and multiple myeloma.

[0051] The above kit, wherein the agent containing irinotecan is a FOLFIRINOX agent, a NALIRIFOX agent, an IRIFOX agent, or an IRINOX agent.

[0052] The aforementioned kit, wherein the agent comprising irinotecan comprises a nanosized particle, a nanosized micelle, a nanosized liposome, or a nanosized exosome.

[0053] The above kit, wherein the agent for inhibiting or suppressing the expression of TGF-β2 and the agent comprising irinotecan are administered contemporaneously, simultaneously, sequentially, or at different times.

[0054] The above kit, wherein the agent for inhibiting or suppressing the expression of TGF-β2 and the agent comprising irinotecan are administered separately or in combination by injection or infusion.

[0055] The above kit, wherein the agent for inhibiting or suppressing the expression of TGF-β2 is a TGF-β2-specific antisense oligonucleotide that is complementary to the TGF-β2 transcript and has a length of 15 to 30 nucleotides.

[0056] The above kit, wherein the agent for inhibiting or suppressing the expression of TGF-β2 is a TGF-β2-specific antisense oligonucleotide that is complementary to TGF-β2 preRNA, pre-mRNA, or mRNA and is 18 to 21 nucleotides in length.

[0057] The above kit, wherein the agent for inhibiting or suppressing expression of TGF-β2 is one or more TGF-β2-specific antisense oligonucleotides complementary to a TGF-β2 transcript, as shown in Table 1.

[0058] The above kit, wherein the TGF-β2-specific antisense oligonucleotide has no more than one or no more than two mismatches compared to the target human TGF-β2.

[0059] The above kit, wherein the TGF-β2 specific antisense oligonucleotide reduces the level of TGF-β2 transcript by at least 60%, or at least 70%, or at least 80%, or at least 90%.

[0060] The above kit, wherein the TGF-β2-specific antisense oligonucleotide reduces the level of any TGF-β1 transcript and the level of any TGF-β3 transcript by less than 10%, or less than 5%, or less than 1%.

[0061] TGF-β2 specific antisense oligonucleotides One or more nucleotides chemically modified as a phosphorothioate internucleoside linkage, a methoxypropylphosphonate internucleoside linkage, an aminophosphoro linkage to a morpholino group, a 2'-OMe ribose group, a 2'-MOE methoxyethyl ribose group, a 2'-4' constrained methoxyethyl bicyclic ribose group, a 2'-4' constrained ethyl bicyclic ribose group, an LNA ribose group, a 2'-F ribose group, or a 5-methylcytosine base The above kit, comprising:

[0062] The above kit, wherein the antisense agent is conjugated to polyethylene glycol, a lipid, or triantennary N-acetyl-galactosamine.

[0063] The aforementioned kit, wherein the agents for inhibiting or suppressing the expression of TGF-β2 comprise a carrier that is sterile water for injection, normal saline, isotonic saline, or a combination thereof, which may be the same or different for each agent.

[0064] The above kit, wherein the agent for inhibiting or suppressing the expression of TGF-β2 is substantially free of excipients.

[0065] The above kit, wherein the agent for inhibiting or suppressing the expression of TGF-β2 is stable for at least 14 days at 37° C. in a carrier that is substantially free of excipients. [Brief explanation of the drawings]

[0066] [Figure 1] Shows highly significant improvement in overall survival for pancreatic cancer patients with reduced TGF-β2. [Figure 2] Shows highly significant improvement in overall survival for pancreatic cancer patients with reduced TGF-β2. [Figure 3] Demonstrates highly significant improvement in overall survival for pancreatic cancer patients with advanced hypoxia. [Figure 4] Pancreatic cancer patients with reduced TMB show highly significant improvement in overall survival. [Figure 5] For pancreatic cancer patients who have TGF-β2 reduced and are treated with irinotecan, we show a highly significant and surprising improvement in overall survival. [Figure 6] For pancreatic cancer patients who have TGF-β2 reduced and are treated with irinotecan, we show a highly significant and surprising improvement in overall survival. [Figure 7] For pancreatic cancer patients who have had TGF-β2 reduced and are treated with the FOFIR irinotecan drug, we show a highly significant and surprising improvement in overall survival. [Figure 8] They showed a highly significant improvement in overall survival for pancreatic cancer patients with reduced TGF-β2, where patients were selected for macrophage reduction. [Figure 9] They showed a highly significant improvement in overall survival for pancreatic cancer patients with reduced TGF-β2, where patients were selected due to a combination of reduced neoantigen load and reduced macrophages measured in the tumor microenvironment. DETAILED DESCRIPTION OF THE INVENTION

[0067] Detailed Description of Disclosure The present invention relates to methods, compositions, agents, and their therapeutic uses for treating or ameliorating the symptoms of cancer in human or animal subjects with agents and compositions designed to promote anti-tumor activity and circumvent drug resistance effects against a variety of different cancers.

[0068] Examples of synergistic pharmaceutical treatment methods of the present invention include combinations of active agents, such as agents for inhibiting or suppressing the expression of TGF-β2, and agents such as irinotecan and formulations containing irinotecan.

[0069] Aspects of the present invention include methods, agents, and their therapeutic uses for treating or ameliorating the symptoms of oncological diseases, wherein the agents may be administered contemporaneously, simultaneously, sequentially, or at separate times.

[0070] In certain embodiments, highly stable formulations of one or more anti-TGF-β2 agents may be used against oncological diseases in combination with one or more irinotecan-containing agents, wherein the anti-TGF-β2 agents and the irinotecan-containing agents are used contemporaneously, simultaneously, sequentially, or at separate times.

[0071] In some embodiments, one or more biomarkers can be used to select subjects who will benefit from a method, agent, or use. Combinations of therapeutic agents disclosed herein, guided by biomarker selection, can result in surprising improvements in overall patient survival.

[0072] The therapeutic compositions of the present disclosure may also be used in combination with chemotherapy and other standard of care therapies.

[0073] In some embodiments, the present invention contemplates the combination of TGF-β2 inhibitors with agents such as irinotecan and irinotecan formulations, which can be guided by biomarkers.

[0074] The present invention recognizes that overexpression of TGF-β2 is a useful indicator for avoiding a series of downstream effects and poor outcomes in oncological diseases.

[0075] In some embodiments, subjects can be selected for therapy based on TGF-β2 as a biomarker to provide improved outcomes. Subjects can be selected if they have elevated expression of TGF-β2.

[0076] In some embodiments, detection of specific biomarkers can be used as a guide to select subjects for therapy involving a combination of a TGF-β2 inhibitor with various anti-cancer drugs.

[0077] In some embodiments, the present invention comprises antisense oligonucleotides as agents that selectively target and inhibit or suppress the expression of TGF-β2 in patients, which are surprisingly superior in oncological disease outcomes to agents that target TGF-β1 or TGF-β3.

[0078] In some embodiments, the combination of a TGF-β2 antisense inhibitor with an irinotecan-based anti-cancer agent and / or other agent can be surprisingly effective.

[0079] Increased efficacy and avoidance of drug resistance effects are characterized by improvements in overall survival (OS) of patients.

[0080] In a further embodiment, a method for treating or ameliorating symptoms of cancer in a human or animal subject in need thereof comprises administering to the subject a therapeutically sufficient amount of a pharmaceutical composition comprising an agent for inhibiting or suppressing expression of TGF-β2, and administering to the subject a therapeutically sufficient amount of a pharmaceutical composition comprising an agent such as irinotecan or a formulation containing irinotecan, wherein the subject may be selected using a TGF-β2 biomarker, and the subject may be selected if expression of TGF-β2 is elevated.

[0081] In an additional embodiment, the present invention includes an antisense agent for inhibiting or suppressing the expression of TGF-β2 in combination with an agent comprising irinotecan for use in treating or ameliorating symptoms of cancer in a subject, wherein the subject is selected if the expression of TGF-β2 is elevated.

[0082] In certain embodiments, the method includes the use of an antisense agent for inhibiting or suppressing expression of TGF-β2 in combination with an agent comprising irinotecan in the preparation of a medicament for treating or ameliorating symptoms of cancer in a subject in need thereof, wherein the subject is selected if expression of TGF-β2 is elevated.

[0083] In some embodiments, the present invention includes a method for treating or ameliorating symptoms of cancer in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of an antisense agent for inhibiting or suppressing expression of TGF-β2, and administering to the subject a therapeutically effective amount of an agent comprising irinotecan, wherein the subject is selected if expression of TGF-β2 is elevated.

[0084] The therapeutic methods of the present invention may be applied to a variety of different cancers, including pancreatic cancer, colorectal cancer, gastric cancer, cervical cancer, lung cancer, melanoma, skin cancer, breast cancer, prostate cancer, kidney cancer, ovarian cancer, thymic cancer, and multiple myeloma.

[0085] Methods and compositions for cancer In many cancers, such as pancreatic cancer and melanoma, patients show high levels of TGF-β2 expression.The agent that inhibits or suppresses the expression of TGF-β2 can be effective in treating these cancer types.For example, in patients with pancreatic cancer, the overall survival time can be more than doubled, from 15 months for patients with high TGF-β2 to 37 months for patients with low TGF-β2.

[0086] The present invention includes a method for treating or ameliorating symptoms of cancer in a human or mammalian subject in need thereof by administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising an agent for inhibiting or suppressing the expression of TGF-β2 in combination with an additional anti-cancer agent, such as irinotecan or an agent containing irinotecan.

[0087] In certain embodiments, the present invention includes an agent for inhibiting or suppressing the expression of TGF-β2 in combination with an agent such as irinotecan or an agent containing irinotecan, e.g., a FOLFIRI or IRIFOX agent, for use in treating or ameliorating symptoms of cancer in a human or mammalian subject.

[0088] The present invention further contemplates the use of a composition comprising an agent for inhibiting or suppressing the expression of TGF-β2 in the preparation of a medicament for treating or ameliorating symptoms of cancer in a subject in combination with an agent such as irinotecan or a formulation containing irinotecan, e.g., a FOLFIRINOX agent, a NALIRIFOX agent, an IRIFOX agent, or an IRINOX agent.

[0089] Therapeutic methods of the present invention using one or more agents to inhibit or suppress the expression of TGF-β2 may be used against pancreatic cancer, colorectal cancer, gastric cancer, cervical cancer, lung cancer, melanoma, skin cancer, breast cancer, prostate cancer, kidney cancer, ovarian cancer, thymic cancer, and multiple myeloma.

[0090] The pharmaceutical compositions of the present disclosure may include an agent for inhibiting or suppressing the expression of TGF-β2 and are administered to a subject in a therapeutically sufficient amount.

[0091] As used herein, the term "agent" can refer to a regimen using one or more active compounds, or a composition containing one or more active compounds and a carrier and / or solvent and / or any number of excipients. In the regimen, the active agents or their formulations may be administered contemporaneously, simultaneously, sequentially, or at different times. In some embodiments, the composition may be a pharmaceutical composition. In certain embodiments, the composition may be a pharmaceutical composition containing a therapeutically effective amount of one or more active compounds. Some examples of excipients are listed 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 a therapeutically effective amount of an agent are known in the art.

[0092] In some embodiments, the present invention comprises an antisense agent for suppressing the expression of TGF-β2 in combination with an agent or regimen comprising irinotecan for use in treating or ameliorating symptoms of cancer in a subject.

[0093] In certain embodiments, the present invention includes an antisense agent for suppressing the expression of TGF-β2 for use in combination with an agent or regimen comprising irinotecan for use in treating or ameliorating symptoms of cancer in a subject.

[0094] As used herein, the term "irinotecan-containing agent" can refer to a regimen that includes irinotecan. An irinotecan-containing agent or regimen can include or employ any form of irinotecan.

[0095] As used herein, the term kit containing "an agent comprising irinotecan" can refer to a kit comprised of active agents or formulations for use in a regimen.

[0096] Human TGF-β2-specific phosphorothioate antisense oligodeoxynucleotide agents Antisense oligonucleotides (ASOs) are single-stranded deoxyribonucleotides that can be complementary to mRNA targets. Antisense therapy can downregulate molecular targets by inducing RNase H endonuclease activity, which cleaves RNA-DNA heteroduplexes, resulting in a significant reduction in target gene translation. Other ASO mechanisms may include inhibition of 5' cap formation, alteration of splicing processes such as splice switching, and steric hindrance of ribosomal activity.

[0097] Antisense therapeutic strategies utilize single-stranded DNA oligonucleotides that inhibit protein production 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 offer a potential therapeutic approach by providing an approach for identifying potential targets.

[0098] Antisense oligonucleotides can be small, synthetic fragments of single-stranded DNA that can be 15-30 nucleotides in length. ASOs specifically bind to complementary DNA / RNA sequences through Watson-Crick hybridization, and upon binding to the target RNA, they can inhibit the translation process either by inducing cleavage mechanisms or by inhibiting mRNA maturation. ASOs can selectively inhibit gene expression with specificity. Chemical modifications of DNA or RNA can be used to enhance stability.

[0099] For example, modifications can be introduced into phosphodiester bonds, sugar rings, and backbones. ASO antiviral agents can block the translation process either by (i) cleaving mRNA via ribonuclease H (RNAse H) or RNase P, or (ii) sterically (non-binding) blocking the enzymes involved in target gene translation. Human TGF-β2-specific phosphorothioate antisense oligodeoxynucleotides, such as OT-101, AP12009 travedelsen SEQ ID NO: 1, can be used to reduce the level of TGF-β2 protein in malignant tumors, thereby slowing disease progression.

[0100] Antisense oligodeoxynucleotides are short strands 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' linkages are modified to phosphorothioates. The molecular formula is C 177 H 208 N 60 Na 17 O 94 P 17 S 17 and has a molecular weight of 6,143 g / mol. OT-101 was designed to be complementary to a specific sequence of human TGF-β2 mRNA following gene expression.

[0101] Antisense oligodeoxynucleotides are short strands of DNA designed to downregulate gene expression by interfering with the translation of specific encoded proteins at the mRNA level.For example, SEQ ID NO:1 (OT-101) is a synthetic 18-mer phosphorothioate oligodeoxynucleotide (S-ODN) in which the non-bridging oxygen of each phosphate moiety is replaced by a sulfur atom.OT-101 is complementary to a specific sequence of human TGF-β2 mRNA from gene expression.OT-101 can be an RNA therapeutic designed to neutralize the immunosuppressive effect of TGF-β2 and reduce the level of TGF-β2 in malignant tumors, so as to treat or improve cancer symptoms or delay the progression of disease.

[0102] The target TGF-β2 mRNA can be the NCBI reference sequence: NM_003238.3, which is 5,882 bp in length. The target region of the TGF-β2 mRNA can be the protein coding sequence from 1,369 to 2,613 of the reference.

[0103] Examples of agents of the present disclosure for inhibiting or suppressing the expression of TGF-β2 include the TGF-β2-specific antisense oligonucleotides set forth in Table 1, SEQ ID NOs: 1-136.

[0104] Table 1. TGF-β2-specific antisense oligonucleotides TIFF2026507184000002.tif29147TIFF2026507184000003.tif235147TIFF20265071840 00004.tif235147TIFF2026507184000005.tif235147TIFF2026507184000006.tif84147

[0105] The sequences in Table 1 can be chemically modified to provide active variants thereof, LNA variants thereof as well as gapmer variants thereof, as known in the art. The sequences in Table 1 can be used in any combination as an active agent, including as a pool combination.

[0106] It is understood that additional antisense oligonucleotides can be constructed based on the TGF-β2 gene sequence.

[0107] In some embodiments, the TGF-β2-specific antisense oligonucleotides of the present invention may have no more than one or no more than two mismatches compared to the target human TGF-β2.

[0108] In certain embodiments, the TGF-β2-specific antisense oligonucleotides of the invention may reduce the level of TGF-β2 transcript by at least 60%, or at least 70%, or at least 80%, or at least 90%.

[0109] In additional embodiments, the TGF-β2-specific antisense oligonucleotides of the present invention may be selective for TGF-β2 and may reduce the levels of any TGF-β1 transcript and any TGF-β3 transcript by less than 10%, or less than 5%, or less than 1%.

[0110] In further embodiments, the therapeutically effective amount of the antisense agent for inhibiting or suppressing expression of TGF-β2 may be 0.1 to 3000 mg / day, or 1 to 1000 mg / day, or 2 to 500 mg / day, or 2 to 200 mg / day.

[0111] In certain embodiments, a formulation of an antisense agent for inhibiting or suppressing expression of TGF-β2 may have a concentration of 0.05 to 50 μM, or 0.1 to 25 μM, or 0.1 to 10 μM, or 0.1 to 7.5 μM, or 0.1 to 5 μM.

[0112] In certain embodiments, the method for using an antisense agent to inhibit or suppress expression of TGF-β2 comprises administering an antisense agent at a dose of 1 to 1000 mg / m 2 / day, or 1–500 mg / m 2 / day, or 1–250 mg / m 2 / day, or 1–100 mg / m 2 / day, or 1–50 mg / m 2 A dose of 1 / day can be used. The average human body surface area is approximately 1.6 to 1.9 m 2 It could be.

[0113] In additional embodiments, the method for using an antisense agent to inhibit or suppress expression of TGF-β2 comprises administering a dose of 0.05 to 40 mg / kg / day, or 0.1 to 30 mg / kg / day, or 0.2 to 20 mg / m 2 / day, or 0.3–10 mg / m 2 / day, or 0.5–5 mg / m 2 A dosage of 100 mg / day can be used. The average human weight can be about 60 kg.

[0114] In certain embodiments, the agent of the present disclosure for inhibiting or suppressing the expression of TGF-β2 may be prepared from a lyophilized powder of the agent.

[0115] In some examples and embodiments, the agent may be a TGF-β2-specific antisense oligonucleotide selected from SEQ ID NOs: 1-136, and may be administered or used by injection or infusion at a dose of 4 μl / min at a dose level of 10 μM on days 1-7, or at a dose of 20 μM on days 1-7, or at a dose of 40 μM on days 1-7, or at a dose of 80 μM on days 1-7. In some embodiments, the agent may be a TGF-β2-specific antisense oligonucleotide selected from SEQ ID NOs: 9-136, and may be administered or used by injection or infusion at a dose of 4 μl / min at a dose level of 10 μM on days 1-7, or at a dose of 20 μM on days 1-7, or at a dose of 40 μM on days 1-7, or at a dose of 80 μM on days 1-7.

[0116] In some examples and embodiments, the agent may be a TGF-β2 gene sequence-specific antisense oligonucleotide selected from SEQ ID NOs: 1-136, and may be administered or used by injection or infusion at a dose of 4 μl / min or 2-8 μl / min, at a dose level of 2 μM on days 1-7, or at a dose of 4 μM on days 1-7, or at a dose of 8 μM on days 1-7, or at a dose of 10 μM on days 1-7. In some embodiments, the agent may be a TGF-β2 gene sequence-specific antisense oligonucleotide selected from SEQ ID NOs: 9-136, and may be administered or used by injection or infusion at a dose of 4 μl / min or 2-8 μl / min, at a dose level of 2 μM on days 1-7, or at a dose of 4 μM on days 1-7, or at a dose of 8 μM on days 1-7, or at a dose of 10 μM on days 1-7.

[0117] Aspects of the invention that involve the administration or use of compositions of agents can ameliorate or suppress symptoms caused by TGF-β2-induced proteins.

[0118] As used herein, the term agent can refer to a single active agent or a composition of a single active agent with a carrier.

[0119] Embodiments of the present invention further include pharmaceutical compositions for inhibiting or suppressing the expression of TGF-β2 or for treating or ameliorating symptoms of cancer in humans or animals. The pharmaceutical composition may include a TGF-β2 inhibitor, artemisinin, a pharmaceutically acceptable salt form, ester, polymorph, or stereoisomer thereof, or any combination thereof, and a carrier. The TGF-β2 inhibitor may be selected from TGF-β2-specific antisense oligonucleotides, such as SEQ ID NOs: 1-136 or SEQ ID NOs: 9-136, and chemically modified variants thereof. The carrier may be sterile water for injection, normal saline, isotonic saline, or a combination thereof.

[0120] Importantly, the composition of the present disclosure may be substantially excipient-free.The composition of the present invention that is substantially excipient-free has been found to be surprisingly stable in carrier.In some embodiments, the composition can be stable in carrier at 37 ℃ for at least 14 days, or at least 21 days, or at least 28 days.

[0121] In additional embodiments, the pharmaceutical composition for injection may comprise less than 1% by weight of the excipient, or less than 0.5% by weight of the excipient, or less than 0.1% by weight of the excipient.

[0122] Embodiments of the present invention further contemplate treatment modalities in which the compositions of the present invention are administered or utilized in combination with standard treatment therapies for a disease. Examples of additional pharmaceutical agents that may be administered or utilized in combination with the compositions of the present invention include anti-inflammatory drugs, anti-inflammatory steroids, piperiquine, pyronaridine, curcumin, frankincense, remdesivir, Sompraz D, Zifi CV / Zac D, CCM, Broclear, Budamate, Rapitas, Montec LC, low molecular weight heparin, prednisolone, paracetamol, vitamin B complex, vitamin C, pantoprozole, doxycycline, ivermectin, zinc, Folacort Rotacaps inhalant, ceftriaxone injectable, paracetamol tablets, Fragmin injectable, Covifor tablets, azithromycin, dexamethasone injectable, odondansetron injectable, multivitamin tablets, ascorbic acid tablets, calcium carbonate tablets, and zinc sulfate tablets.

[0123] Some TGF-β2 specific antisense oligonucleotide agents are set forth in US 9,963,703, US 9,758,786, and US 8,476,246.

[0124] For example, the API travedersen (OT-101) is a synthetic 18-mer S-ODN composed of the bases adenine (A), thymine (T), guanine (G), and cytosine (C), with all 3'-5' linkages modified to phosphorothioate. This sulfur modification makes the drug more resistant to degradation, resulting in improved stability in vitro and in vivo. Its primary molecular structure, the nucleotide sequence, was designed to be complementary to a specific sequence in human transforming growth factor beta 2 (TGF-β2) mRNA. This sequence and related sequences can be used due to their excellent chemical and structural properties, biological activity, and specificity to achieve the highest antisense efficacy in vitro and in vivo.

[0125] The chemical structure of travedersen, an example of the phosphorothioate moiety (CAG), and physical properties are shown in Table 2.

[0126] Table 2. Chemical and physical properties of travedelsen TIFF2026507184000007.tif83162

[0127] The investigational medicinal product may be supplied as a sterile lyophilizate for solution for infusion in a 50 mL glass vial (primary container) containing 7.37 mg of travedersen (intratumoral treatment) and a 20R glass vial (primary container) containing 250 mg of travedersen (intravenous treatment). The finished medicinal product may contain no excipients. The glass vials may be used for parenteral administration. The glass vials may be sealed with sterile rubber stoppers suitable for lyophilization. The stoppers may be sealed with crimped capsules containing colored flip-off caps. For clinical use, each vial may be provided in a white, collapsible box to protect the vials from light exposure and damage during transport. Both the glass vial and the collapsible box may be labeled according to local requirements. The primary as well as secondary containers in the closed system may meet international quality standards for packaging of sterile solid pharmaceutical products for injection. The kit supplies OT-101 as a lyophilized powder in various amounts in 50 mL glass vials, and the total volume (mL) and resulting concentration (µM) after dissolution can be specified.

[0128] The present invention further provides a kit containing lyophilized powder of one or more TGF-β2-specific antisense oligonucleotides selected from SEQ ID NOs: 1-136 in vials, each containing 250 mg. The kit may also contain suitable vials and all necessary components of an application system, i.e., syringes, tubing, and filters. The OT-101 lyophilized powder can be dissolved in isotonic (0.9%) aqueous sodium chloride solution before use.

[0129] Anticancer drug and therapeutic drug combinations Embodiments of the present invention include combinations of TGF-β2 inhibitors with anti-cancer agents such as irinotecan and formulations or regimens containing irinotecan.

[0130] Examples of irinotecan-containing formulations, agents, and / or regimens include FOLFIRINOX, NALIRIFOX, IRIFOX, and IRINOX. The dosage and time of administration of such irinotecan-containing formulations, agents, and / or regimens are known in the art.

[0131] The irinotecan-containing agents of the present disclosure can be formulated in various forms, such as small particle form, micelle form, hydrogel form, liposome form, or exosome form.

[0132] In certain embodiments, the present invention provides a therapeutic combination of one or more antisense TGF-β2 inhibitors and an agent containing irinotecan with a nanosized particle, nanosized micelle, nanosized hydrogel, nanosized liposome, or nanosized exosome.

[0133] Additional aspects of the present invention include therapeutic combinations of TGF-β2 inhibitors with FOLFIRINOX, NALIRIFOX, IRIFOX, or IRINOX agents in conjunction with standard of care therapy.

[0134] The unexpectedly beneficial synergistic effects of therapeutic combinations for cancer treatment of antisense TGF-β2 inhibitors and irinotecan-containing agents may be amplified if patients are selected based on biomarkers for clinical activity.

[0135] In the treatment of cancer patients, the therapeutic combination of the agent that contains irinotecan combined with antisense TGF-β2 inhibitor can significantly increase the overall survival time of patients.In some embodiments, the therapeutic combination of the agent that contains irinotecan with antisense TGF-β2 inhibitor can provide unexpectedly advantageous synergistic effect based on clinical data.

[0136] Synergistic effects of therapeutic combinations Embodiments of the present invention can provide a synergistic therapeutic combination of an agent for inhibiting or suppressing the expression of TGF-β2 and an agent containing a topoisomerase I inhibitor for treating or ameliorating symptoms of cancer.

[0137] Examples of irinotecan agents include irinotecan, irinotecan hydrochloride, irinotecan sucrose sulfate, and irinotecan sucrose sulfate forms.

[0138] Examples of topoisomerase I inhibitors include irinotecan, topotecan, rubitecan, and camptothecin derivatives and analogs such as 9-aminocamptothecin.

[0139] In some embodiments, the anti-cancer use of the combination of an antisense agent for inhibiting or suppressing the expression of TGF-β2 and an agent containing irinotecan can be particularly effective for patients with high levels of certain biomarkers, such as TGF-β2 in particular. Biomarkers can be used to select patients who will benefit from combination therapy.

[0140] Examples of regimens using irinotecan-containing agents include FOLFIRINOX, NALIRIFOX, IRIFOX, and IRINOX, as known in the art. For example, NALIRIFOX is liposomal irinotecan formulated with oxaliplatin, 5-fluorouracil, and leucovorin. For example, IRINOX is a combination of irinotecan formulated with oxaliplatin. For example, FOLFIRINOX is a combination of irinotecan hydrochloride formulated with leucovorin calcium (folinic acid), fluorouracil, and oxaliplatin. For example, FOLFIRI is a formulation of folinic acid (leucovorin, calcium folinate, or FA), fluorouracil (5FU), and irinotecan. For example, IRINOX is a formulation of irinotecan and oxaliplatin.

[0141] A drawback of conventional therapy using only irinotecan-containing agents is that not all cancer patients are suitable for such treatment. Irinotecan-containing agents can be used for cancer patients with low TGF-β2 levels, which indicate a low risk of progression. However, cancer patients with high TGF-β2 levels, which indicate a risk of progression, and / or other biomarker levels that suggest a risk of progression are typically treated only with palliative therapy, such as gemcitabine.

[0142] Embodiments of the present invention that combine an agent for inhibiting or suppressing TGF-β2 expression with an agent containing irinotecan may show the surprising advantage that overall survival and / or HRQoL may be improved for all cancer patients.

[0143] For example, embodiments of the invention utilizing a combination of a TGF-β2 inhibitor and an agent containing irinotecan can be used in cancer patients with low TGF-β2.

[0144] Additionally, embodiments of the present invention can be used in cancer patients with elevated TGF-β2 and / or other biomarker levels indicative of a risk of progression.

[0145] Therefore, the embodiment of the present invention that combines an antisense agent for inhibiting or suppressing the expression of TGF-β2 with an agent containing irinotecan can be used advantageously for all cancer patients.

[0146] Irinotecan-containing agents can be used against cancers including pancreatic cancer, colorectal cancer, gastric cancer, cervical cancer, lung cancer, melanoma, skin cancer, breast cancer, prostate cancer, kidney cancer, ovarian cancer, thymic cancer, and multiple myeloma.

[0147] Embodiments of the present invention can provide a therapeutic combination of an agent for inhibiting or suppressing TGF-β2 expression with an agent containing a topoisomerase I inhibitor to treat or ameliorate symptoms of cancer, including pancreatic cancer, colorectal cancer, gastric cancer, cervical cancer, lung cancer, melanoma, skin cancer, breast cancer, prostate cancer, kidney cancer, ovarian cancer, thymic cancer, and multiple myeloma. The topoisomerase I inhibitor can be irinotecan, irinotecan hydrochloride, irinotecan sucrose sulfate, irinotecan sucrose sulfate, FOLFIRINOX agents, NALIRIFOX agents, IRIFOX agents, IRINOX agents, topotecan, rubitecan, or 9-aminocamptothecin.

[0148] Numbered aspects of the present disclosure include the following: (1) An antisense agent for inhibiting or suppressing the expression of TGF-β2, in combination with an agent comprising irinotecan, for use in treating or ameliorating symptoms of cancer in a subject. (2) Use of an antisense agent for inhibiting or suppressing the expression of TGF-β2 in the preparation of a medicament for treating or ameliorating symptoms of cancer in a subject in combination with an agent comprising irinotecan. (3) 1. A method for treating or ameliorating a symptom of cancer in a subject in need thereof, comprising: administering to the subject a therapeutically effective amount of an antisense agent to inhibit or suppress expression of TGF-β2; and administering to said subject a therapeutically effective amount of an agent comprising irinotecan. The method comprising: (4) The agent, use, or method of any of Aspects 1 to 3, wherein the agent comprising irinotecan is a FOLFIRINOX agent, a NALIRIFOX agent, an IRIFOX agent, or an IRINOX agent. (5) The agent, use, or method of any of embodiments 1-4, wherein said agent comprising irinotecan comprises a nanosized particle, a nanosized micelle, a nanosized liposome, or a nanosized exosome. (6) The agent, use, or method of any of aspects 1 to 5, wherein the agent for inhibiting or suppressing expression of TGF-β2 and the agent comprising irinotecan are administered contemporaneously, simultaneously, sequentially, or at separate times. (7) The agent, use, or method of any of aspects 1 to 6, wherein the agent for inhibiting or suppressing expression of TGF-β2 and the agent comprising irinotecan are administered separately or in combination by injection or infusion. (8) The agent, use, or method of any of aspects 1-7, wherein the cancer is pancreatic cancer, colorectal cancer, gastric cancer, cervical cancer, lung cancer, melanoma, skin cancer, breast cancer, prostate cancer, kidney cancer, ovarian cancer, thymic cancer, and multiple myeloma. (9) The agent, use, or method of any of aspects 1 to 8, wherein the agent for inhibiting or suppressing expression of TGF-β2 is a TGF-β2-specific antisense oligonucleotide that is complementary to a TGF-β2 transcript and is 15 to 30 nucleotides in length. (10) The agent, use, or method of any of aspects 1 to 9, wherein the agent for inhibiting or suppressing expression of TGF-β2 is a TGF-β2-specific antisense oligonucleotide that is complementary to TGF-β2 pre-RNA, pre-mRNA, or mRNA and is 18 to 21 nucleotides in length. (11) 11. The agent, use, or method of any of embodiments 1 to 10, wherein said agent for inhibiting or suppressing expression of TGF-β2 is one or more TGF-β2-specific antisense oligonucleotides complementary to the TGF-β2 transcripts in Table 1, and chemically modified variants thereof, LNA variants thereof, gapmer variants thereof, and any combination or pool thereof. (12) 12. The agent, use, or method of any of aspects 1-11, wherein said TGF-β2-specific antisense oligonucleotide has no more than one or no more than two mismatches compared to the target human TGF-β2. (13) 13. The agent, use, or method of any of embodiments 1-12, wherein said TGF-β2-specific antisense oligonucleotide reduces the level of TGF-β2 transcript by at least 60%, or at least 70%, or at least 80%, or at least 90%. (14) 14. The agent, use, or method of any of embodiments 1-13, wherein the TGF-β2-specific antisense oligonucleotide reduces the level of any TGF-β1 transcript and the level of any TGF-β3 transcript by less than 10%, or less than 5%, or less than 1%. (15) TGF-β2 specific antisense oligonucleotides One or more nucleotides chemically modified as a phosphorothioate internucleoside linkage, a methoxypropylphosphonate internucleoside linkage, an aminophosphoro linkage to a morpholino group, a 2'-OMe ribose group, a 2'-MOE methoxyethyl ribose group, a 2'-4' constrained methoxyethyl bicyclic ribose group, a 2'-4' constrained ethyl bicyclic ribose group, an LNA ribose group, a 2'-F ribose group, or a 5-methylcytosine base 15. The agent, use, or method of any one of aspects 1 to 14, comprising: (16) 16. The agent, use, or method of any of embodiments 1-15, wherein said antisense agent is conjugated to polyethylene glycol, a lipid, or triantennary N-acetyl-galactosamine. (17) 17. The agent, use, or method of any of Aspects 1 to 16, wherein the agent for inhibiting or suppressing expression of TGF-β2 comprises a carrier that is sterile water for injection, saline, isotonic saline, or a combination thereof, which may be the same or different for each agent. (18) 18. The agent, use, or method of any of aspects 1 to 17, wherein said agent for inhibiting or suppressing expression of TGF-β2 is substantially excipient-free. (19) 19. The agent, use, or method of any of aspects 1 to 18, wherein the agent for inhibiting or suppressing expression of TGF-β2 is stable in a substantially excipient-free carrier at 37°C for at least 14 days. (20) The agent, use, or method of any of embodiments 1-19, wherein the patient is selected to have been previously treated with any agent including irinotecan. (twenty one) 21. The agent, use, or method of any of embodiments 1-20, comprising using one or more biomarkers to select subjects who will benefit from said agent, use, or method. (twenty two) 22. The agent, use, or method of any of embodiments 1 to 21, wherein the patient is selected to have elevated levels of TGF-β2. (twenty three) 23. The agent, use, or method of any of aspects 1-22, wherein said one or more biomarkers is a level of tumor mutational burden (TMB), a level of tumor neoantigens, a level of clinical hypoxia, or a combination thereof. (twenty four) 24. The agent, use, or method of any of aspects 1-23, wherein said one or more biomarkers are the level of tumor mutational burden, the level of reduced neoantigen load measured in the tumor microenvironment, the level of reduced macrophages measured in the tumor microenvironment, the level of reduced mesenchymal stem cells (MSCs) measured in the tumor microenvironment, the level of type 2 helper T cells (Th2) measured in the tumor microenvironment, and combinations thereof. (twenty five) 25. The method, agent, or use of any of embodiments 1-24, wherein a subject receiving said administration or use has an improved level of at least one of said one or more biomarkers compared to levels found in a healthy patient. (26) 26. The agent, use, or method of any of embodiments 1-25, wherein said administration or use reduces mortality at 6, 12, 18, 24, 30, or 36 months. (27) 27. The agent, use or method of any of embodiments 1-26, wherein said administration or use increases survival at 6, 12, 18, 24, 30, or 36 months. (28) 28. The agent, use or method of any of embodiments 1-27, wherein said administration or use improves health-related quality of life (HRQoL) by 15%, or 25%, or 35%, or 45%. (29) The agent, use, or method of any of embodiments 1-28 in combination with any one or more medicaments including cancer-targeting agents, cancer growth blockers, EGFR inhibitors, and combinations thereof. (30) 30. The agent, use, or method of any of embodiments 1-29 in combination with any one or more medicaments selected from bevacizumab, everolimus, velzutifan, dabrafenib, trametinib, and combinations thereof. (31) 31. The agent, use, or method of any of embodiments 1-30, in combination with any one or more pharmaceutical agents that are cancer growth blockers selected from angiogenesis inhibitors, histone deacetylase inhibitors, hedgehog blockers, mTOR inhibitors, p53 inhibitors, PARP inhibitors, proteasome inhibitors, tyrosine kinase inhibitors, and combinations thereof. (32) The agent, use, or method of any of embodiments 1-31, in combination with any one or more pharmaceutical agents that are EGFR inhibitors selected from erlotinib, gefitinib, afatinib, osimertinib, dacomitinib, and combinations thereof. (33) The agent, use, or method of any of embodiments 1 to 32 in combination with a standard of care treatment for cancer. (34) The agent, use, or method of any of embodiments 1-33 in combination with radiation therapy or electric field therapy. (35) a therapeutically effective amount of an antisense agent for inhibiting or suppressing the expression of TGF-β2, and Therapeutically effective amount of an agent comprising irinotecan A kit for treating or ameliorating a symptom of cancer, comprising: (36) The kit of embodiment 35, wherein the cancer is pancreatic cancer, colorectal cancer, gastric cancer, cervical cancer, lung cancer, melanoma, skin cancer, breast cancer, prostate cancer, kidney cancer, ovarian cancer, thymic cancer, and multiple myeloma. (37) The kit of any of Aspects 35 to 36, wherein the agent comprising irinotecan is a FOLFIRINOX agent, a NALIRIFOX agent, an IRIFOX agent, or an IRINOX agent. (38) The kit of any of embodiments 35 to 37, wherein the agent comprising irinotecan comprises a nanosized particle, a nanosized micelle, a nanosized liposome, or a nanosized exosome. (39) The kit of any of aspects 35 to 38, wherein the agent for inhibiting or suppressing expression of TGF-β2 and the agent comprising irinotecan are administered contemporaneously, simultaneously, sequentially, or at different times. (40) The kit of any of aspects 35 to 39, wherein the agent for inhibiting or suppressing expression of TGF-β2 and the agent comprising irinotecan are administered separately or in combination by injection or infusion. (41) 41. The kit of any of Aspects 35 to 40, wherein the agent for inhibiting or suppressing expression of TGF-β2 is a TGF-β2-specific antisense oligonucleotide that is complementary to a TGF-β2 transcript and has a length of 15 to 30 nucleotides. (42) 42. The kit of any of Aspects 35 to 41, wherein the agent for inhibiting or suppressing expression of TGF-β2 is a TGF-β2-specific antisense oligonucleotide that is complementary to TGF-β2 preRNA, pre-mRNA, or mRNA and is 18 to 21 nucleotides in length. (43) The kit of any of aspects 35 to 42, wherein the agent for inhibiting or suppressing expression of TGF-β2 is one or more TGF-β2-specific antisense oligonucleotides complementary to a TGF-β2 transcript, as shown in Table 1. (44) 44. The kit of any of aspects 35 to 43, wherein the TGF-β2-specific antisense oligonucleotide has no more than one or no more than two mismatches compared to the target human TGF-β2. (45) 45. The kit of any of embodiments 35 to 44, wherein the TGF-β2-specific antisense oligonucleotide reduces the level of TGF-β2 transcript by at least 60%, or at least 70%, or at least 80%, or at least 90%. (46) 46. ​​The kit of any of embodiments 35 to 45, wherein the TGF-β2-specific antisense oligonucleotide reduces the level of any TGF-β1 transcript and the level of any TGF-β3 transcript by less than 10%, or less than 5%, or less than 1%. (47) The TGF-β2-specific antisense oligonucleotide One or more nucleotides chemically modified as a phosphorothioate internucleoside linkage, a methoxypropylphosphonate internucleoside linkage, an aminophosphoro linkage to a morpholino group, a 2'-OMe ribose group, a 2'-MOE methoxyethyl ribose group, a 2'-4' constrained methoxyethyl bicyclic ribose group, a 2'-4' constrained ethyl bicyclic ribose group, an LNA ribose group, a 2'-F ribose group, or a 5-methylcytosine base 47. The kit of any one of aspects 35 to 46, comprising: (48) 48. The kit of any of embodiments 35 to 47, wherein the antisense agent is conjugated to polyethylene glycol, a lipid, or triantennary N-acetyl-galactosamine. (49) 49. The kit of any of Aspects 35 to 48, wherein the agents for inhibiting or suppressing expression of TGF-β2 comprise a carrier that is sterile water for injection, saline, isotonic saline, or a combination thereof, which may be the same or different for each agent. (50) The kit of any one of Aspects 35 to 49, wherein the agent for inhibiting or suppressing the expression of TGF-β2 is substantially free of excipients. (51) The kit of any of Aspects 35 to 50, wherein the agent for inhibiting or suppressing the expression of TGF-β2 is stable at 37° C. for at least 14 days in a carrier that is substantially free of excipients.

[0149] All publications, including patents, published patent applications, and non-patent publications, as well as the sequence listing, referenced herein are each expressly incorporated herein by reference in their entirety for all purposes.

[0150] While the foregoing disclosure has been described in detail by way of example for purposes of clarity of understanding, it will be apparent to those skilled in the art that certain changes and modifications are encompassed by the present disclosure and can be made without undue experimentation within the scope of the appended claims, which are presented by way of example and not limitation. The present invention encompasses all such additional embodiments, equivalents, and modifications. The present invention includes any combination or mixture of the features, materials, elements, or limitations of the various exemplary components, examples, and claimed embodiments.

[0151] It is emphasized that, according to common practice, the features of the drawings are of any scale herein and are intended to cover similar features which may be enlarged or reduced at will. [Example]

[0152] Example 1 The disclosed methods and agents for suppressing TGF-β2 can be used for pancreatic cancer.

[0153] A clinical outcome study of patients with pancreatic cancer showed that decreased TGF-β2 expression was associated with improved overall survival (OS). This result was surprising because neither the TGF-β1 nor the TGF-β3 isoform was significantly associated with increased survival in pancreatic cancer. Therefore, this clinical study supports TGF-β2 as a sole therapeutic target in pancreatic cancer.

[0154] A Kaplan-Meier Plotter was used to calculate patient survival and determine the association between improved overall survival and reduced TGF-β2 expression in pancreatic cancer.

[0155] The results of the study on pancreatic cancer are shown in Figure 1. Figure 1 shows a highly significant (log-rank P=0.00515) improvement in overall survival for pancreatic cancer patients with reduced TGF-β2. As understood in the art, a log-rank p-value of less than about 0.05 (or <0.1) confirms that the difference is significant. The improvement found in overall survival with reduced TGF-β2 expression was a significant increase from 16 months in the TGF-β2 high expression cohort to 37 months in the TGF-β2 low expression cohort.

[0156] The study in Figure 1 used quartile values ​​of TGFB2 expression from the Z-score of mRNA expression (log2 RSEM-UQ). Quartile statistics are shown in Table 3. Overall survival and other survival calculations are shown in Table 4.

[0157] Table 3. Quartile statistics of TGF-β2 for pancreatic cancer TIFF2026507184000008.tif69145

[0158] Table 4. Overall and other survival calculations for TGF-β2 in pancreatic cancer TIFF2026507184000009.tif33128

[0159] Example 2. The disclosed methods and agents for suppressing TGF-β2 can be used for pancreatic cancer.

[0160] Clinical outcome studies on pancreatic cancer patients have been conducted and have shown improved overall survival (OS) with decreased TGF-β2 expression. Additional inclusion criteria: For this study, patients who had not received follow-up cancer therapy after surgery were selected.

[0161] A Kaplan-Meier Plotter was used to calculate patient survival and determine the association between improved overall survival and reduced TGF-β2 expression in pancreatic cancer.

[0162] The results of the pancreatic cancer study are shown in Figure 2, which shows a highly significant (log-rank P=0.0111) improvement in overall survival for pancreatic cancer patients with reduced TGF-β2. The improvement in overall survival found with reduced TGF-β2 expression was a significant increase from 7 months in the high TGF-β2 expression cohort to at least 14 months in the low TGF-β2 expression cohort (B).

[0163] For the study in Figure 2, TGFB2 quartile values ​​from the z-scores of mRNA expression for all samples were used (log RNA Seq V2 RSEM). Quartile statistics are shown in Table 5. Overall survival and other survival calculations are shown in Table 6.

[0164] Table 5. Quartile statistics of TGF-β2 for pancreatic cancer TIFF2026507184000010.tif69145

[0165] Table 6. Overall survival and other survival calculations for TGF-β2 agents in pancreatic cancer TIFF2026507184000011.tif25128

[0166] Example 3. Clinical outcome studies have been conducted on pancreatic cancer patients and have shown improved overall survival (OS) with reduced hypoxia, so patients can be selected based on hypoxia as a biomarker.

[0167] A Kaplan-Meier Plotter was used to calculate patient survival and determine the association between improved overall survival and reduced hypoxia in pancreatic cancer.

[0168] The results of the pancreatic cancer study are shown in Figure 3, which shows a highly significant (log-rank P=0.003126) improvement in overall survival for pancreatic cancer patients in whom hypoxia was reduced. The improvement found in overall survival with reduced hypoxia was a significant increase from 15 months in the high hypoxia cohort to 30 months in the low hypoxia cohort.

[0169] The study in Figure 3 used quartiles of the Buffa Hypoxia Score as a biomarker for patient selection. Quartile statistics are shown in Table 7. Overall survival and other survival calculations are shown in Table 8.

[0170] Table 7. Hypoxia quartile statistics for pancreatic cancer TIFF2026507184000012.tif69145

[0171] Table 8: Overall survival and other survival calculations for pancreatic cancer TIFF2026507184000013.tif33128

[0172] Example 4. Clinical outcome studies on pancreatic cancer patients have shown that a reduction in tumor mutation burden (TMB) is associated with improved overall survival (OS). Therefore, patients can be selected based on tumor mutation burden (TMB) as a biomarker.

[0173] A Kaplan-Meier Plotter was used to calculate patient survival and determine the association between improved overall survival and reduced TMB in pancreatic cancer.

[0174] The results of the pancreatic cancer study are shown in Figure 4, which shows a highly significant (log-rank P=0.003013) improvement in overall survival for pancreatic cancer patients with reduced TMB. The improvement found in overall survival with reduced TMB was a significant increase from 16 months in the high TMB cohort to at least 22 months in the low TMB cohort (B).

[0175] The study in Figure 4 used quartiles of TMB (non-synonymous) as a biomarker to select patients. Quartile statistics are shown in Table 9. Overall survival and other survival calculations are shown in Table 10.

[0176] Table 9. Quartile statistics of TMB for pancreatic cancer TIFF2026507184000014.tif69145

[0177] Table 10: Overall survival and other survival calculations for pancreatic cancer TIFF2026507184000015.tif25128

[0178] Example 5. The disclosed methods and agents for inhibiting TGF-β2 can be used for pancreatic cancer in combination with treatment with the agent irinotecan.

[0179] A clinical outcome study of patients with pancreatic cancer showed improved overall survival (OS) with reduced TGF-β2 expression, where patients were selected based on treatment with irinotecan. This result was surprising because improved OS with reduced TGF-β2 expression was not observed when patients were also treated with gemcitabine alone, fluorouracil alone, oxaliplatin alone, or leucovorin alone.

[0180] A Kaplan-Meier Plotter was used to calculate patient survival and determine the association between improved overall survival and reduced TGF-β2 expression in pancreatic cancer when patients were also treated with irinotecan.

[0181] The results of the pancreatic cancer study are shown in Figure 5, which shows a highly significant (log-rank P=0.0174) improvement in overall survival for pancreatic cancer patients with reduced TGF-β2. The improvement found in overall survival with reduced TGF-β2 expression was a significant increase from 23 months in the high TGF-β2 expression cohort to a much longer period of 34 months in the low TGF-β2 expression cohort. The significant increase in overall survival indicates, among other things, that drug resistance effects are being avoided.

[0182] These results in Figure 5 demonstrate a surprising synergistic effect on increasing overall survival in treating pancreatic cancer using a therapy that combines reducing TGF-β2 expression with an agent containing irinotecan. These results were particularly surprising because none of oxaliplatin, leucovorin, or fluorouracil showed significant synergistic effects in combination with reducing TGF-β2 expression.

[0183] For the study in Figure 5, TGFB2 quartile values ​​from z-scores of mRNA expression for all samples were used (log RNA Seq V2 RSEM). Quartile statistics are shown in Table 11. Overall survival and other survival calculations are shown in Table 12.

[0184] Table 11. Quartile statistics of TGF-β2 for pancreatic cancer TIFF2026507184000016.tif44145

[0185] Table 12. Overall and other survival calculations for TGF-β2 in pancreatic cancer TIFF2026507184000017.tif25128

[0186] Example 6 The disclosed methods and agents for inhibiting TGF-β2 can be used for pancreatic cancer in combination with treatment with the agent irinotecan.

[0187] Clinical outcome studies have been conducted on pancreatic cancer patients, showing improved overall survival (OS) with decreased TGF-β2 expression, where patients were selected based on treatment with irinotecan.

[0188] Patient survival was calculated using a Kaplan-Meier Plotter to determine the association between improved overall survival and reduced TGF-β2 expression in pancreatic cancer when patients were also treated with irinotecan.

[0189] The results of the pancreatic cancer study are shown in Figure 6, which shows a highly significant (log-rank P=0.0152) improvement in overall survival for pancreatic cancer patients with reduced TGF-β2. The improvement found in overall survival with reduced TGF-β2 expression was a significant increase from 8 months in the high TGF-β2 expression cohort to at least 30 months in the lower TGF-β2 expression cohort (C). The significant increase in overall survival indicates, among other things, that drug resistance effects are avoided.

[0190] TGFB2 quartile values ​​from z-scores of mRNA expression for all samples were used in the study in Figure 6 (log RNA Seq V2 RSEM). Quartile statistics are shown in Table 13. Overall survival calculations are shown in Table 14.

[0191] Table 13. Quartile statistics of TGF-β2 for pancreatic cancer TIFF2026507184000018.tif69145

[0192] Table 14. Overall survival calculations for TGF-β2 in pancreatic cancer TIFF2026507184000019.tif18128

[0193] Example 7 The disclosed methods and agents for inhibiting TGF-β2 can be used for pancreatic cancer in combination with treatment with the agent irinotecan.

[0194] Clinical outcome studies have been conducted on patients with pancreatic cancer, showing improved overall survival (OS) with decreased TGF-β2 expression, in which patients were selected based on treatment with irinotecan-containing FOLFIRINOX.

[0195] A Kaplan-Meier plotter was used to calculate patient survival and determine the association between improved overall survival and reduced TGF-β2 expression in pancreatic cancer when patients were also treated with irinotecan-containing FOLFIRINOX.

[0196] The results of the pancreatic cancer study are shown in Figure 7. Figure 7 shows a significant (log-rank P=0.0952) improvement in overall survival for pancreatic cancer patients with reduced TGF-β2. The improvement found in overall survival with reduced TGF-β2 expression was a significant increase from 23 months in the high TGF-β2 expression cohort to at least 34 months in the lower TGF-β2 expression cohort (B). The significant increase in overall survival indicates, among other things, that drug resistance effects are avoided.

[0197] The study in Figure 7 used quartile values ​​of TGF-β2 from z-scores of mRNA expression for all samples (log RNA Seq V2 RSEM). Quartile statistics are shown in Table 15. Overall survival calculations are shown in Table 16.

[0198] Table 15. Quartile statistics of TGF-β2 for pancreatic cancer TIFF2026507184000020.tif44145

[0199] Table 16. Overall survival calculations for TGF-β2 in pancreatic cancer TIFF2026507184000021.tif25128

[0200] Example 8 The disclosed methods and agents for inhibiting TGF-β2 can be used for pancreatic cancer, where biomarkers can be used to select patients who will benefit from the agents.

[0201] Clinical outcome studies have been conducted on patients with pancreatic cancer, and have shown that reduced TGF-β2 expression is associated with improved overall survival (OS), using biomarkers to select patients who will benefit from the agent. Significant biomarkers for pancreatic cancer include reduced tumor mutation burden, reduced neoantigen load measured in the tumor microenvironment, reduced macrophages measured in the tumor microenvironment, reduced mesenchymal stem cells (MSCs) measured in the tumor microenvironment, and increased type 2 helper T cells (Th2) measured in the tumor microenvironment. Any combination of these biomarkers can also be used.

[0202] A Kaplan-Meier Plotter was used to calculate patient survival and determine the association between improved overall survival and reduced TGF-β2 expression in pancreatic cancer, where biomarkers were used to select patients who would benefit from the agent.

[0203] The results of the pancreatic cancer study are shown in Figure 8. Figure 8 shows a significant (log-rank P = 0.0025) improvement in overall survival for pancreatic cancer patients with reduced TGF-β2, where patients were selected using reduced macrophages measured in the tumor microenvironment. The improvement in overall survival observed with reduced TGF-β2 expression and reduced macrophages was a significant increase from 15 months in the high TGF-β2 expression cohort to 73 months in the low TGF-β2 expression cohort. This significant increase in overall survival indicates, among other things, that drug resistance effects are being avoided.

[0204] Similar results were obtained with p-values ​​for additional biomarkers as follows: reduced tumor mutational burden (log-rank P = 0.079), reduced neoantigen burden measured in the tumor microenvironment (log-rank P = 0.051); Decreased mesenchymal stem cells (MSCs) measured in the tumor microenvironment (log-rank P=0.031); Increased type 2 helper T cells (Th2) measured in the tumor microenvironment (log-rank P = 0.025); The combination of reduced tumor mutational burden and reduced M0 macrophages (log-rank P=0.0018), and A combination of reduced neoantigen load measured in the tumor microenvironment and reduced M0 macrophages (see Figure 9, log-rank P=0.000076).

Claims

1. An antisense agent for inhibiting or suppressing the expression of TGF-β2, in combination with an agent comprising irinotecan, for use in treating or ameliorating symptoms of cancer in a subject.

2. Use of an antisense agent for inhibiting or suppressing the expression of TGF-β2 in the preparation of a medicament for treating or ameliorating symptoms of cancer in a subject in combination with an agent comprising irinotecan.

3. 1. A method for treating or ameliorating a symptom of cancer in a subject in need thereof, comprising: administering to the subject a therapeutically effective amount of an antisense agent to inhibit or suppress expression of TGF-β2; and administering to said subject a therapeutically effective amount of an agent comprising irinotecan. The method comprising:

4. The agent, use, or method according to any one of claims 1 to 3, wherein the agent comprising irinotecan is a FOLFIRINOX agent, a NALIRIFOX agent, an IRIFOX agent, or an IRINOX agent.

5. 5. The agent, use, or method according to any one of claims 1 to 4, wherein the agent comprising irinotecan comprises a nanosized particle, a nanosized micelle, a nanosized liposome, or a nanosized exosome.

6. The agent, use, or method of any one of claims 1 to 5, wherein the agent for inhibiting or suppressing expression of TGF-β2 and the agent comprising irinotecan are administered contemporaneously, simultaneously, sequentially, or at separate times.

7. 7. The agent, use, or method according to any one of claims 1 to 6, wherein the agent for inhibiting or suppressing expression of TGF-β2 and the agent comprising irinotecan are administered separately or in combination by injection or infusion.

8. 8. The agent, use, or method according to any one of claims 1 to 7, wherein the cancer is pancreatic cancer, colorectal cancer, gastric cancer, cervical cancer, lung cancer, melanoma, skin cancer, breast cancer, prostate cancer, kidney cancer, ovarian cancer, thymic cancer, and multiple myeloma.

9. 9. The agent, use, or method according to any one of claims 1 to 8, wherein the agent for inhibiting or suppressing expression of TGF-β2 is a TGF-β2-specific antisense oligonucleotide complementary to the TGF-β2 transcript and having a length of 15 to 30 nucleotides.

10. 10. The agent, use, or method according to any one of claims 1 to 9, wherein the agent for inhibiting or suppressing the expression of TGF-β2 is a TGF-β2-specific antisense oligonucleotide that is complementary to TGF-β2 pre-RNA, pre-mRNA, or mRNA and is 18 to 21 nucleotides in length.

11. The agent for inhibiting or suppressing the expression of TGF-β2 is selected from the following (Table 1):

11. The agent, use, or method of any one of claims 1 to 10, wherein the antisense oligonucleotides are one or more TGF-β2 specific antisense oligonucleotides complementary to the TGF-β2 transcript, and chemically modified variants thereof, LNA variants thereof, gapmer variants thereof, and any combination or pool thereof.

12. 12. The agent, use, or method of claim 11, wherein the TGF-β2-specific antisense oligonucleotide has no more than one or no more than two mismatches compared to the target human TGF-β2.

13. 12. The agent, use, or method of claim 11, wherein the TGF-β2-specific antisense oligonucleotide reduces the level of TGF-β2 transcript by at least 60%, or at least 70%, or at least 80%, or at least 90%.

14. 12. The agent, use, or method of claim 11, wherein the TGF-β2-specific antisense oligonucleotide reduces the levels of any TGF-β1 transcript and any TGF-β3 transcript by less than 10%, or less than 5%, or less than 1%.

15. TGF-β2 specific antisense oligonucleotides One or more nucleotides chemically modified as a phosphorothioate internucleoside linkage, a methoxypropylphosphonate internucleoside linkage, an aminophosphoro linkage to a morpholino group, a 2'-OMe ribose group, a 2'-MOE methoxyethyl ribose group, a 2'-4' constrained methoxyethyl bicyclic ribose group, a 2'-4' constrained ethyl bicyclic ribose group, an LNA ribose group, a 2'-F ribose group, or a 5-methylcytosine base The agent, use or method of claim 11, having the formula:

16. 12. The agent, use, or method of claim 11, wherein the antisense agent is conjugated to polyethylene glycol, a lipid, or triantennary N-acetyl-galactosamine.

17. 17. The agent, use, or method of any one of claims 1 to 16, wherein the agent for inhibiting or suppressing expression of TGF-β2 comprises a carrier that is sterile water for injection, normal saline, isotonic saline, or a combination thereof, which may be the same or different for each agent.

18. 18. The agent, use or method of any one of claims 1 to 17, wherein the agent for inhibiting or suppressing expression of TGF-β2 is substantially excipient-free.

19. 19. The agent, use, or method of any one of claims 1 to 18, wherein the agent for inhibiting or suppressing expression of TGF-β2 is stable in a substantially excipient-free carrier at 37°C for at least 14 days.

20. 20. The agent, use or method of any one of claims 1 to 19, wherein patients are selected who have been previously treated with any agent including irinotecan.

21. 21. The agent, use or method according to any one of claims 1 to 20, comprising using one or more biomarkers to select subjects who will benefit from said agent, use or method.

22. 22. The agent, use or method according to any one of claims 1 to 21, wherein patients having elevated levels of TGF-β2 are selected.

23. 23. The agent, use or method of any one of claims 1 to 22, wherein said one or more biomarkers are levels of tumor mutational burden (TMB), levels of tumor neoantigens, levels of clinical hypoxia, or a combination thereof.

24. 24. The agent, use or method of any one of claims 1 to 23, wherein said one or more biomarkers are the level of tumor mutational burden, the level of reduced neoantigen load measured in the tumor microenvironment, the level of reduced macrophages measured in the tumor microenvironment, the level of reduced mesenchymal stem cells (MSCs) measured in the tumor microenvironment, the level of type 2 helper T cells (Th2) measured in the tumor microenvironment, and combinations thereof.

25. 25. The method, agent or use of any one of claims 1 to 24, wherein a subject receiving said administration or use has an improved level of at least one of said one or more biomarkers compared to levels found in healthy patients.

26. 26. The agent, use or method of any one of claims 1 to 25, wherein said administration or use reduces mortality at 6, 12, 18, 24, 30 or 36 months.

27. 27. The agent, use or method of any one of claims 1 to 26, wherein said administration or use increases survival at 6, 12, 18, 24, 30 or 36 months.

28. 28. The agent, use or method of any one of claims 1 to 27, wherein said administration or use improves health-related quality of life (HRQoL) by 15%, or 25%, or 35%, or 45%.

29. 29. The agent, use or method of any one of claims 1 to 28 in combination with any one or more medicaments including cancer-targeting agents, cancer growth blockers, EGFR inhibitors, and combinations thereof.

30. 30. The agent, use, or method of any one of claims 1 to 29 in combination with any one or more medicaments selected from bevacizumab, everolimus, velzutifan, dabrafenib, trametinib, and combinations thereof.

31. 31. The agent, use, or method of any one of claims 1 to 30 in combination with any one or more medicaments that are cancer growth blockers selected from angiogenesis inhibitors, histone deacetylase inhibitors, hedgehog blockers, mTOR inhibitors, p53 inhibitors, PARP inhibitors, proteasome inhibitors, tyrosine kinase inhibitors, and combinations thereof.

32. 32. The agent, use or method of any one of claims 1 to 31 in combination with any one or more pharmaceutical agents that are EGFR inhibitors selected from erlotinib, gefitinib, afatinib, osimertinib, dacomitinib, and combinations thereof.

33. 33. The agent, use or method of any one of claims 1 to 32 in combination with standard therapeutic treatment for cancer.

34. 34. The agent, use or method of any one of claims 1 to 33 in combination with radiation therapy or electric field therapy.

35. a therapeutically effective amount of an antisense agent for inhibiting or suppressing the expression of TGF-β2, and Therapeutically effective amount of an agent comprising irinotecan A kit for treating or ameliorating a symptom of cancer, comprising:

36. 36. The kit of claim 35, wherein the cancer is pancreatic cancer, colorectal cancer, gastric cancer, cervical cancer, lung cancer, melanoma, skin cancer, breast cancer, prostate cancer, kidney cancer, ovarian cancer, thymic cancer, and multiple myeloma.

37. The kit according to any one of claims 35 to 36, wherein the agent comprising irinotecan is a FOLFIRINOX agent, a NALIRIFOX agent, an IRIFOX agent, or an IRINOX agent.

38. The kit of any one of claims 35 to 37, wherein the agent comprising irinotecan comprises a nanosized particle, a nanosized micelle, a nanosized liposome, or a nanosized exosome.

39. The kit of any one of claims 35 to 38, wherein the agent for inhibiting or suppressing expression of TGF-β2 and the agent comprising irinotecan are administered contemporaneously, simultaneously, sequentially, or at separate times.

40. 40. The kit of any one of claims 35 to 39, wherein the agent for inhibiting or suppressing expression of TGF-β2 and the agent comprising irinotecan are administered separately or in combination by injection or infusion.

41. The kit of any one of claims 35 to 40, wherein the agent for inhibiting or suppressing expression of TGF-β2 is a TGF-β2-specific antisense oligonucleotide complementary to a TGF-β2 transcript and having a length of 15 to 30 nucleotides.

42. The kit of any one of claims 35 to 41, wherein the agent for inhibiting or suppressing expression of TGF-β2 is a TGF-β2-specific antisense oligonucleotide that is complementary to TGF-β2 preRNA, pre-mRNA, or mRNA and is 18 to 21 nucleotides in length.

43. 43. The kit of any one of claims 35 to 42, wherein the agent for inhibiting or suppressing expression of TGF-β2 is one or more TGF-β2-specific antisense oligonucleotides complementary to a TGF-β2 transcript as shown in Table 1.

44. 44. The kit of any one of claims 35 to 43, wherein the TGF-β2-specific antisense oligonucleotide has no more than one or no more than two mismatches compared to the target human TGF-β2.

45. 45. The kit of any one of claims 35 to 44, wherein the TGF-β2-specific antisense oligonucleotide reduces the level of TGF-β2 transcript by at least 60%, or at least 70%, or at least 80%, or at least 90%.

46. 46. ​​The kit of any one of claims 35 to 45, wherein the TGF-β2-specific antisense oligonucleotide reduces the level of any TGF-β1 transcript and the level of any TGF-β3 transcript by less than 10%, or less than 5%, or less than 1%.

47. The TGF-β2-specific antisense oligonucleotide One or more nucleotides chemically modified as a phosphorothioate internucleoside linkage, a methoxypropylphosphonate internucleoside linkage, an aminophosphoro linkage to a morpholino group, a 2'-OMe ribose group, a 2'-MOE methoxyethyl ribose group, a 2'-4' constrained methoxyethyl bicyclic ribose group, a 2'-4' constrained ethyl bicyclic ribose group, an LNA ribose group, a 2'-F ribose group, or a 5-methylcytosine base 47. The kit of any one of claims 35 to 46, comprising:

48. 48. The kit of any one of claims 35 to 47, wherein the antisense agent is conjugated to polyethylene glycol, a lipid, or triantennary N-acetyl-galactosamine.

49. 49. The kit of any one of claims 35 to 48, wherein the agents for inhibiting or suppressing expression of TGF-β2 comprise a carrier that is sterile water for injection, normal saline, isotonic saline, or a combination thereof, which may be the same or different for each agent.

50. 50. The kit of any one of claims 35 to 49, wherein the agent for inhibiting or suppressing expression of TGF-β2 is substantially excipient-free.

51. 51. The kit of any one of claims 35 to 50, wherein the agent for inhibiting or suppressing expression of TGF-β2 is stable in a substantially excipient-free carrier at 37°C for at least 14 days.

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