HMGA-BINDING NUCLEIC ACIDS

摘要

#CMT# #/CMT# Use of L-nucleic acid (I) as an intracellular active substance. #CMT# : #/CMT# Independent claims are included for: (1) a procedure for binding an intracellular receptor comprising providing a cell containing at least one intracellular receptor; providing (I); and incubating the cell with (I); (2) a composition comprising (I) and a discharge vehicle; (3) a high mobility group-A (HMGA) binding nucleic acid, comprising a section box A1 and a section box A2, where the section box A1 and A2 are connected together by intermediate section, and particularly comprises GGGCG, GGGUG or GGGAG; (4) a procedure to screen a HMGA-antagonist or HMGA-agonist comprising either providing a candidate-HMGA-antagonist and/or a candidate HMGA-agonist, providing a nucleic acid; providing a test system, which delivers a signal in the presence of HMGA-antagonist and/or HMGA-agonist, and determining whether the candidate-HMGA-antagonist and/or HMGA-agonist; or providing a phase, particularly solid phase immobilized HMGA, providing nucleic acid, introducing a candidate-HMGA-agonist and/or HMGA-antagonist, and determining; (5) a kit to proof HMGA, comprises the nucleic acid; (6) a HMGA-antagonist and HMGA-agonist, are obtained by the process; (7) a complex comprises a HMGA protein and the nucleic acid; and (8) a nucleic acid binding to a transcriptions factor comprises an AT-hooks. #CMT#ACTIVITY : #/CMT# Cytostatic; Virucide; Antiarteriosclerotic; Antiinflammatory. #CMT#MECHANISM OF ACTION : #/CMT# High mobility group-A (HMGA) antagonist; HMGA-agonist. The affinity of (I) to bind with HMGA was tested using biological assays. The results showed that (I) exhibited a median inhibitory concentration (IC 5 0) value of 15 nM. #CMT#USE : #/CMT# (I) is useful in the preparation of medicament to treat and/or prevent disease such as tumor disease (mesenchymal tumor, epithelial tumor, benign tumor, malignant tumor and metastasis tumor), virus infection and arteriosclerosis (arteriosclerosis plaque), where the cell molecule of the medicament is intracellular cell molecule; disease due to molecular cell HMGA such as carcinoma of prostate, pancreas, thyroid gland, cervix, stomach, breast, colon/rectum and ovarian, neuroblastoma, lymphoma, uterusleiomyoma, lipoma, endometriumpolyps, chondroid hamartoma of lung, pleomorphic adenoma of head salivary gland, hamangiopericytoma, chondromatosis tumor, aggressive agiomyxome, diffuse astrocytoma, osteoclastoma, skin cancer, Burkett's lymphoma, Lewis-lung cancer, leukemia, non-small cell lung carcinoma and/or metastasis. (I) is useful in the preparation of diagnostic agent (all claimed). #CMT#ADVANTAGE : #/CMT# (I) binds comprehensively to the AT-hook transcription factor. #CMT#BIOLOGY : #/CMT# Preferred Process: (I) interacts with intracellular receptor. The incubation step is carried out under a condition, so that (I) binds to the intracellular receptor of the cell. (I) reacts with the intracellular receptor within the cell. The incubating step determines whether (I) binds with the intracellular cell. The discharge vehicle is desirable for intracellular discharge of (I). The discharge vehicle comprises vehicle, conjugate and physical measures. The discharge vehicle is liposome, nanoparticle, microparticle, cyclodextrin or dendrimere, or a vesicle such as polypeptide, polyethylenimine (preferred) and/or amphipathic molecules. The conjugate is receptor-mediated endocytosis, a conjugate with a fusogen peptide, a conjugate with a signal peptide or a conjugate with a nucleic acid, preferably a conjugate with Spiegelmer (RTM: mirror image of oligonucleotide) or a lipophilic conjugate. The determining step is carried out by a test, whether the candidate HMGA-agonist or the candidate HMGA-antagonist replaces the nucleic acid. Preferred Components: (I) is a Spiegelmer (RTM: mirror image of oligonucleotide). The intracellular receptor are: molecular receptor, enzymes, chaperon-molecule, signal peptide, intracellular structures or metabolic intermediate; polypeptide, carbohydrate, nucleic acid and/or lipids; transcriptions factor and AT-hook containing DNA binding proteins; and HMG-proteins, such as HMGA1, HMGA1a, HMGA1b or HMGA2. The intracellular receptor comprises metabolic intermediates and enzymes. The section Z1 is arranged between 6-7 nucleotide and the section Z2 is arranged between 12-25 nucleotide. The nucleic acid exhibits a first section at the 5'-end of the section box A1 and a second section at the 3'-end of the section box A2, where both the sections preferably comprises 4-8 nucleotide or nucleotide pairs and are at least partially or completely hybridized together. The nucleic acid exhibits 5'-end of the section box A1 at the section helix A1 and a 3'-end of the section box A2 at the section helix A2 and comprises 4-8 nucleotide and is partially or completely hybridized together. The 3'-5'-end of the section helix A1 and section box A1 are arranged between 0-5 nucleotide, and 3'-5'-end of the section helix A2 and section box A2 are arranged between 8 nucleotide. The first section is a helix section A1 and the second section is a helix section A2, where both the section comprises 4-8 nucleotide, and the length of section helix A1 is independent of the length of the section helixA2. The section helix B1 is arranged between the 3'-end of the section helix A1 and 5'-end of the section box A1 and the section B2 is arranged between the 3'-end of the section box A2 and the 5'-end of the section helix A2 and where the length of the section helix B1 and B2 comprises 4-8 nucleotide pair. The 3'-end of the section helix A1 and 5'-end of the section helix B1 are arranged between 0-5, preferably 2 nucleotide. The 3'-end of the section helix B2 and the 5'-end of the section helix A2 are arranged between 0-6, preferably 1 nucleotide. The amount of nucleotide in section helix A1, A2, B1 and B2 are 10-12 nucleotide. The amount of hybridized nucleotide in section helix A1, A2, B1 and B2 are 10-12 nucleotide pair. The nucleic acid comprises neither section helix A1, and A2 at 5'-end and section helix B1 and B2 at 3'-end, where the length of section helix B1 and B2 comprises 4-8 nucleotide. The 3'-end of the section helix B1 and the 5'-end of the section box A1 are arranged between 2 nucleotide and the 3'-end of the section box A2 and the 5'-end of the section helix B2 are arranged between 1-7 nucleotide. The section Z1 exhibits sequence N 1N 2GN 8N 3N 4N 5. The nucleic acid comprises a section box A1 and section box A2, where the 3'-end of the section box A1 directly connected with the 5'-end of the section Z1 and the 3'-end of the Z1 directly connected with the 5'-end of the section box A2. The nuclei acid comprises a section helix B1 and B2 and comprise 4-8 nucleotide, that are particularly completely or partially hybridized together. The 5'-3' two nucleotide N 6, N 7 are arranged between the 3'-end of the section helix B1 and 5'-end of the section box A1. The nucleotide sequence GN y is arranged between the 3'-end of the section box A2 and the 5'-end of the section helix B2 or in 5'-3'-directed nucleotide sequence. The nucleic acid comprises section helix A1 and helix A2 and are completely or partially hybridized together. The nucleotide sequence N x is arranged between 3'-end of the section helix A1 and 5'-end of the section helix B1. The nucleotide sequence N z is arranged between 3'-end of the section helix B2 and the 5'-end of the section helix A2. The amount of hybridized nucleotide is 10-12 nucleotide pair, by hybridizing section helix A1 with section helix A2 and section helix B1 with section helix B2. The HMGA binding nucleic acid comprises nucleic acid structure of Helix A 1-N x-Helix B1-N 6N 7BoxA1N 1N 2GN 8N 3N 4N 5BOX A2G-N y-Helix B2-N z-Helix A2, or Helix A 1-N x-Helix B1-N 6N 7BoxA1N 1N 2GN 8N 3N 4N 5BOX A2Helix B2-N z-Helix A2, or Helix B1-N 6N 7BoxA1N 1N 2GN 8N 3N 4N 5BOX A2G-N y-Helix B2, or Helix B1-N 6N 7BoxA1N 1N 2GN 8N 3N 4N 5BOX-Helix B2, or HelixA1-N a-Box A1-N b-Helix C1-N c-Helix C2-N d-Box A 2-G-N e-Helix A2; or Helix A1-N iHelix B1-N j-Box A1-A-Helix C1-N c-Helix C2-G-Box A2-G-Helix B2-A-Helix A2. The nucleotide sequence comprises: fully defined 47-50 nucleic acid (SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, or SEQ ID NO: 13) sequence, as given in the specification; fully defined 48 nucleic acid (SEQ ID NO: 3), as given in the specification; or fully defined 33 nucleic acid (SEQ ID NO: 15 and SEQ ID NO: 16) sequence, as given in the specification; or fully defined 50 nucleic acid (SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 14, SEQ ID NO: 22 or SEQ ID NO: 24) sequence as given in the specification; or fully defined 50 nucleic acid (SEQ ID NO: 12) sequence as given in the specification. The 3'-end of the section box A2 connects directly to the 5'-end of the section helix B2. The section Z2 comprises section helix C1 and C2, which is arranged at central section N c. The length of the section helix C1 and helix C2 is either greater or equal, or 3-6, preferably 3-5 nucleotide. The sections helix C1 and helix C2 are together completely or partially hybridized. The nucleic acid comprises a section box A1 and a section box A2, where the nucleotide sequence N b is arranged between the 3'-end of the section box A1 and the 5'-end of the section helix C1 and comprises 3 nucleotide. The nucleic acid comprises section box A1 and section box A2, where the nucleotide sequence is arranged between the 3'-end of the section helix C2 and the 5'-end of the section box A2. The nuclei acid comprises a section helix A1 and a section helix A2, where the section comprises 5-6 nucleotide and particularly completely or partially together hybridized. The nucleotide sequence N a is arranged between the 3'-end of the section helix A1 and the 5'-end of the section box A1. The nucleotide sequence GN e is arranged between the 3