Modified antisense oligonucleotide for inhibition of FoxP3 expression
The present invention refers to an oligonucleotide comprising 12 to 25 nucleotides, wherein at least one of the nucleotides comprises a modification selected from the group consisting of a bridged nucleic acid such as LNA, ENA, a 2′Fluoro modified nucleotide, a 2 O-Methyl modified nucleotide, a 2 O-Methoxy modified nucleotide, a FANA and a combination thereof. The oligonucleotide hybridizes with a nucleic acid sequence of Foxp3 of SEQ ID NO. 1 and/or of SEQ ID NO. 2 resulting in a reduction of the expression of FoxP3 mRNA, FoxP3 pre-mRNA or a combination thereof. The invention is further directed to a pharmaceutical composition comprising an oligonucleotide of the present invention and to the oligonucleotide and pharmaceutical composition, respectively for use in a method of preventing and/or treating a disorder, where FoxP3 imbalance is involved.
1 . An oligonucleotide comprising the nucleotide sequence of SEQ ID NO:81, wherein at least one of the nucleotides comprises a modification selected from the group consisting of LNA (locked nucleic acid), ENA (2′-O,4′-C-ethylene-bridged nucleic acid), a 2′-fluoro modified nucleotide, a 2-O-methyl modified nucleotide, a 2-O-methoxy modified nucleotide, a FANA (2′-deoxy-2-fluoro-D-arabinonucleic acid), and a combination thereof, and hybridizing with a nucleic acid sequence of Foxp3 of SEQ ID NO. 1 and/or of SEQ ID NO. 2 resulting in a reduction of FoxP3, FoxP3 mRNA, FoxP3 pre-mRNA or a combination thereof of 40% to 99% within 6 to 240 h or within 12 to 120 h from first administration of the oligonucleotide compared to an untreated control.
2 . The oligonucleotide according to claim 1 , wherein the oligonucleotide results in a reduction of FoxP3, FoxP3 mRNA, FoxP3 pre-mRNA, or a combination thereof, of 40% to 99%, within 24 to 72 h from first administration of the oligonucleotide to a subject.
3 . The oligonucleotide according to claim 1 , wherein the oligonucleotide hybridize with Foxp3 of SEQ ID NO. 1 and/or SEQ ID NO. 2, wherein the oligonucleotide hybridizes within a region of position 1510 to 2109 of SEQ ID NO. 2.
4 . The oligonucleotide according to claim 1 , wherein the oligonucleotide is
(A25126H; SEQ ID NO. 81)
+G*+A*+A*G*T*A*A*T*C*T*G*T*G*C*G*+A*+G*+C,
wherein + indicates an LNA modified nucleotide and * indicates a phosphorothioate (PTO) linkage between the nucleotides.
5 . The oligonucleotide according to claim 1 , wherein the oligonucleotide inhibits the expression of FoxP3, FoxP3 mRNA, FoxP3 pre-mRNA or a combination thereof at a nanomolar or micromolar concentration.
6 . A pharmaceutical composition comprising an oligonucleotide according to claim 1 and a pharmaceutically acceptable carrier, excipient, diluent or a combination thereof.
7 . The pharmaceutical composition of claim 6 , further comprising an antibody.
8 . The pharmaceutical composition of claim 7 , wherein the antibody inhibits expression or activity of PD-1.
9 . The pharmaceutical composition of claim 7 , wherein the antitumor active agent is an antibody that inhibits expression or activity of a factor involved in cancer progression and/or metastasis selected from the group consisting of SND1, MTDH, HER-2, BRAF, KRAS, VEGF, EGFR1, EGFR2, BCR/ABL, ABL, MET, ALK, JAK2, BTK, miR-223, CCL18, CCL20, Lcn2, CCL5/CCR9, DDR2, PHD2, IL6, SDF-1/CXCL12 and a combination thereof.
10 . The oligonucleotide according to claim 1 that reduces expression of FoxP3, FoxP3 mRNA, FoxP3 pre-mRNA or a combination thereof when administered to a subject having a disorder characterized by overexpression of FoxP3, FoxP3 mRNA, FoxP3 pre-mRNA.
11 . The oligonucleotide according to claim 10 , wherein the disorder is a malignant and/or benign tumor, a chronic infectious disease, a chronic inflammatory disease caused by infection or a combination thereof.
12 . The oligonucleotide according to claim 11 , wherein the malignant tumor is selected from the group consisting of breast cancer, lung cancer, malignant melanoma, lymphoma, skin cancer, bone cancer, prostate cancer, liver cancer, brain cancer, cancer of the larynx, gall bladder cancer, pancreatic cancer, testicular cancer, rectal cancer, parathyroid cancer, thyroid cancer, adrenal cancer, neural tissue cancer, head and neck cancer, colon cancer, stomach cancer, bronchial cancer, kidney cancer, basal cell carcinoma, squamous cell carcinoma, metastatic skin carcinoma, osteo sarcoma, Ewing's sarcoma, reticulum cell sarcoma, liposarcoma, myeloma, giant cell tumor, small-cell lung tumor, islet cell tumor, primary brain tumor, meningioma, acute and chronic lymphocytic and granulocytic tumors, acute and chronic myeloid leukemia, hairy-cell tumor, adenoma, hyperplasia, medullary carcinoma, intestinal ganglioneuromas, Wilm's tumor, seminoma, ovarian tumor, leiomyomater tumor, cervical dysplasia, retinoblastoma, soft tissue sarcoma, malignant carcinoid, topical skin lesion, rhabdomyosarcoma, Kaposi's sarcoma, osteogenic sarcoma, malignant hypercalcemia, renal cell tumor, polycythemia vera, adenocarcinoma, anaplastic astrocytoma, glioblastoma multiforma, leukemia, epidermoid carcinoma and a combination thereof.
13 . The oligonucleotide according to claim 11 , wherein the chronic infectious disease is selected from the group consisting of hepatitis B and/or C virus, human immune deficiency virus, cytomegalovirus, Herpes Simplex virus, measles virus, respiratory syncytial virus, Helicobacter pylori infection and a combination thereof, or wherein the chronic inflammatory disease caused by infection is selected from the group consisting of chronic inflammatory diseases of the liver, liver fibrosis, liver cirrhosis and a combination thereof.
14 . The pharmaceutical composition according to claim 11 , wherein the composition is suitable to be administered locally or systemically.