US-12622936-B2 - Anti-tumor composition

US12622936B2US 12622936 B2US12622936 B2US 12622936B2US-12622936-B2

Abstract

The present invention relates to an oncolytic adenovirus capable of co-expressing interleukin-12 and a C-met-inhibiting oligonucleotide; and an antitumor immune-boosting composition and anticancer composition comprising the oncolytic adenovirus. The present invention has first identified an adenovirus system having simultaneous effects of IL-12 expression and C-met inhibition in cancer gene treatment. The adenovirus system of the present invention is capable of inhibiting C-met while expressing interleukin-12, thereby restoring immune functions in a tumor environment to enhance anticancer effects such as the inhibition of tumor recurrence and tumor growth and to inhibit tumor migration. Accordingly, the adenovirus system of the present invention can be effectively used in the treatment of cancer.

Inventors

Chae Ok Yun
Eon Ju OH

Assignees

GENEMEDICINE CO., LTD.

Dates

Publication Date: 20260512
Application Date: 20190710
Priority Date: 20180710

Claims (13)

1 . A recombinant oncolytic adenovirus comprising a first heterologous gene and a second heterologous gene; wherein the first heterologous gene encodes interleukin-12 (IL-12); and wherein the second heterologous gene encodes an oligonucleotide that is complementary to mRNA of C-met and inhibits expression of C-met, wherein the oligonucleotide that is complementary to mRNA of C-met is selected the group consisting of small hairpin RNA (shRNA), siRNA, antisense oligonucleotides, and guide RNA for CRISPR.
2 . The recombinant oncolytic adenovirus according to claim 1 , wherein the first heterologous gene encoding interleukin-12 (IL-12) comprises an IL-12A (p35) gene sequence and an IL-12B (p40) gene sequence.
3 . The recombinant oncolytic adenovirus according to claim 1 , wherein the first heterologous gene encoding interleukin-12 (IL-12) further comprises a linker sequence or internal ribosome entry site (IRES) sequence between the IL-12A (p35) gene sequence and the IL-12B (p40) gene sequence.
4 . The recombinant oncolytic adenovirus according to claim 1 , wherein the first heterologous gene encoding interleukin-12 (IL-12) is inserted into an E1B region of the recombinant oncolytic adenovirus.
5 . The recombinant oncolytic adenovirus according to claim 1 , wherein the second heterologous gene expressing an oligonucleotide that is complementary to mRNA of C-met is inserted into an E1B region of the recombinant oncolytic adenovirus.
6 . The recombinant oncolytic adenovirus according to claim 1 , wherein, when the oligonucleotide complementarily binding to the C-met gene to inhibit expression of C-met is a guide RNA for CRISPR, wherein the recombinant oncolytic adenovirus further comprises a nucleotide sequence expressing Cas protein.
7 . The recombinant oncolytic adenovirus according to claim 1 , further comprising a biocompatible polymer in a form of being combined with the recombinant oncolytic adenovirus.
8 . The recombinant oncolytic adenovirus according to claim 7 , wherein the biocompatible polymer is selected from the group consisting of PEI-Arg-mPEG-S-S-mPEG-Arg-PEI (PAPS), mPEG-PEI-g-Arg-S-S-Arg-g-PEI-mPEG (PPSA), pegylated and iron oxide nanoparticles-crosslinked catechol-grafted poly L lysine (PICION), an arginine-grafted biodegradable polymer (ABP), a pegylated and PTX-conjugated polymeric micelle (APP), mPEG-b-Pip-CBA (PPCBA), PPCBA-PEI-Arginine (PPA), poly(amidoamine) dendrimer (PAMAM), polyethyleneglycol (PEG), poly-lactide (PLA), polyglycolide (PGA), poly-lactide-co-glycolide (PLGA), poly-ε-caprolactone (PCL), polyethylenimine (PEI), hyaluronic acid (HA), gelatin, chitosan, and serum albumin.
9 . A method for treating cancer, comprising administering a therapeutically effective amount of an adenovirus system according to claim 1 to a subject.
10 . The method for treating cancer according to claim 9 , wherein the cancer is gastric cancer, lung cancer, non-small cell lung cancer, breast cancer, ovarian cancer, liver cancer, bronchial cancer, nasopharyngeal cancer, laryngeal cancer, pancreatic cancer, bladder cancer, colorectal cancer, colon cancer, cervical cancer, bone cancer, non-small cell bone cancer, blood cancer, skin cancer (melanoma etc.), head or neck cancer, uterine cancer, rectal cancer, anal cancer, colon cancer, fallopian tube cancer, endometrial cancer, vaginal cancer, vulvar cancer, Hodgkin's disease, esophageal cancer, small intestine cancer, endocrine cancer, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, prostate cancer, chronic or acute leukemia, lymphocytic lymphoma, kidney or ureter cancer, renal cell carcinoma, renal pelvic carcinoma, polyploid carcinoma, salivary gland cancer, sarcoma cancer, pseudomyxoma, hepatoblastoma, testicular cancer, glioblastoma, cleft lip cancer, ovarian germ cell tumor, basal cell carcinoma, multiple myeloma, gallbladder cancer, choroidal melanoma, ampulla of vater cancer, peritoneal cancer, adrenal cancer, tongue cancer, small cell cancer, pediatric lymphoma, nerve blastoma, duodenal cancer, ureteral cancer, astrocytoma, meningioma, renal carcinoma, vulvar cancer, thymus cancer, central nervous system (CNS) tumor, primary central nervous system lymphoma, spinal cord tumor, brain stem glioma, or pituitary adenoma.
11 . The method for treating cancer according to claim 9 , wherein the composition is intratumorally administered.
12 . A method for enhancing antitumor immunity, comprising administering a therapeutically effective amount of an adenovirus system according to claim 1 to a subject.
13 . The method for enhancing antitumor immunity according to claim 12 , wherein the composition is intratumorally administered.

Description

TECHNICAL FIELD The present invention relates to a composition for antitumor or enhancing antitumor immunity comprising a recombinant adenovirus that co-expresses interleukin-12 (IL-12) and a nucleic acid molecule inhibiting an expression of a tyrosine kinase Met (C-met). BACKGROUND ART Tumor immunotherapy is a method of inducing an immune response against a tumor by enhancing the overall immune function of the body and treating tumors through the same. Research on tumor immunotherapy is actively being conducted. However, an immunosuppressive environment is almost always formed at the stage at which cancer is generated, so that, even if the body's immune system is actively working, it is difficult to easily remove the tumor. Tumor cells themselves express many abnormal antigens. These antigens elicit an eradication reaction through immune surveillance, allowing tumor cells to evade immune surveillance even if the body's immune system is active. In addition, it has been found that the phenomenon is mediated by various factors produced by tumor cells. It has been reported that tumor tissues can produce immunosuppressive molecules such as vascular endothelial growth factor (VEGF), tumor growth factor (TGF)-β and interleukin (IL)-10, and regulatory T cells penetrate the immune-suppressed tumor. Further, stimulation of the expression of an inhibitory receptor called PD-1 continuously increases by activated T cells, and in the case of cancer cells, PD-L1, which is a ligand specifically binding to PD-1, is expressed in a large amount to inactivate activated T cells to avoid immune responses, resulting in an immune suppression microenvironment and immune tolerance in tumors. A recent study has proved that an inhibitor capable of inhibiting the immune checkpoint of PD-1, which is involved in suppressing T cell activity, induces a strong antitumor immune response. Consequently, overcoming immune surveillance avoidance is a major strategy for immunotherapy. Therefore, as a way to overcome these limitations, research into directly introducing a cytokine gene having an immune-promoting effect into cancer cells to generate and secrete cytokines from cancer cells, and thus, specifically eliminating cancer cells through induction of antitumor immune responses is actively underway. Among the immune-enhancing cytokine genes for which antitumor effects have been reported to date, interleukin-12 (IL-12) is one of the most effective and promising cytokines. Interleukin-12 (IL-12), which is a heterodimeric protein containing 40 kDa and 35 kDa subunits linked by disulfide bonds, is secreted from antigen presenting cells (APCs) such as activated macrophages, monocytes, dendritic cells and activated B lymphocytes, and acts directly on T cells and NK cells that can effectively eliminate cancer cells to activate the T cells and NK cells and induce the secretion of IFN-γ, as well as enhance the killing ability of the T cells and NK cells against cancer cells. Local expression of IL-12 makes tumor cells sensitive to T-cell mediated cytotoxicity, resulting in inhibition of tumor growth and establishment of systemic immunity. However, there is a difficulty in the clinical application of IL-12 because the administration of IL-12 can cause systemic cytokine-associated toxicity that limits the patient's acceptable dose. In addition, overall downregulation of the immune effect, increased IL-10 expression in the patient's serum, and IL-12 polarization from Th1 to Th2 immunity due to decreased IFN-γ and TNF-β expression occur. For this reason, IL-12 is sometimes repeatedly administered. These clinical results show the limitations of IL-12 as a single treatment for the treatment of cancer. Recently, studies on growth factors and receptors thereof as biological indicators reflecting the degree of malignant cancer are underway. Thereamong, studies on C-met, which is a member of the tyrosine kinase receptor family, are being actively conducted. C-met acts as a receptor for hepatocyte growth factor/scatter factor (HGF/SF), and is overexpressed in various cancers. Most patients with C-met overexpression have a poor cancer treatment prognosis. The overexpression of C-met enhances mitogenesis and cellular motility due to the HGF/SF-Met signaling system, inhibits apoptosis, forms blood vessels, and induces invasion and migration into the extracellular matrix (ECM), i.e., causes an increase in cancer malignancy. Since rapid proliferation and migration of cancer cells induced by overexpression of C-met exceed the rate of removal of cancer cells in the immune system, it is difficult to eliminate the cancer cells through an immune response. Furthermore, since the center of the immune response in a tumor microenvironment is inclined severely toward inhibition as the size of the tumor increases, it is more difficult to control tumor cells using a single therapy, and the improved antitumor effect by antitumor immunotherapy cannot be achieved. Therefore, the d