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Nucleic acid aptamers targeting lymphocyte activation gene 3 (lag-3) and uses thereof

A nucleic acid aptamer and lymphocyte technology, which is applied in the fields of genetic engineering, medical preparations containing active ingredients, organic active ingredients, etc.

Pending Publication Date: 2021-04-13
ONENESS BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, only a fraction of cancer patients have been found to respond to these treatments

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  • Nucleic acid aptamers targeting lymphocyte activation gene 3 (lag-3) and uses thereof
  • Nucleic acid aptamers targeting lymphocyte activation gene 3 (lag-3) and uses thereof
  • Nucleic acid aptamers targeting lymphocyte activation gene 3 (lag-3) and uses thereof

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example

[0158] So that the invention described herein may be more fully understood, the following examples are set forth. The examples described in this application are provided to illustrate the systems and methods provided herein and are not to be construed in any way as limiting the scope thereof.

example 1

[0159] Example 1 : Identification and Characterization of the LAG-3 Aptamer

[0160] Candidate lymphocyte activation gene 3 (LAG-3) aptamers were identified from a synthetic ssDNA library using a high-throughput SELEX assay targeting human recombinant LAG-3. Such LAG-3 aptamer candidates were subjected to next-generation sequencing and tested for their activity in disrupting the interaction of LAG-3 with MHC-II using the LAG-3 / MHC-II bioassay. Several LAG-3 aptamers were found to disrupt the interaction between LAG-3 and MHC-II, as indicated by the expression of a luciferase reporter gene. exist Figure 1A Exemplary results are provided in . An anti-LAG-3 antibody was used as a positive control. Figure 1A .

[0161] LAG-3 aptamers B4, B8, D9 and F4 in Figure 1A Marked by arrows in , the sequences of these aptamers are provided in Table 1 below. Sequence alignment revealed conserved motifs among the identified LAG-3 aptamers, such as Figure 1B shown. Aptamers inclu...

example 2

[0172] Example 2 : Synthesis and Characterization of the LAG-3 Aptamer in Tetrameric Form

[0173] The tetrameric form of the LAG-3 aptamer was constructed using two backbone sequences with complementary segments such that they can anneal together by base pairing. Each backbone sequence can be linked to two aptamers at the 5' and 3' ends, thereby forming an aptamer tetramer. This ligation is achieved using a backbone sequence having at each end a primer sequence complementary to a primer sequence in the aptamer sequence that allows the aptamer to base pair to each end of the backbone sequence. Exemplary aptamer sequences and backbone sequences are shown in Table 3.

[0174] Table 3. Exemplary backbone and aptamer sequences.

[0175]

[0176] *Primer sequences are underlined. P16 indicates a 16-residue primer, and P10 indicates a 10-residue primer.

[0177] The backbone and aptamer sequences were mixed in various molar ratios, and the tetramers thus formed were separated...

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Abstract

Nucleic acid aptamers capable of binding to lymphocyte activation gene 3 (LAG-3) and uses thereof for modulating immune responses. Such aptamers may comprise a G-rich motif, for example, GX1GGGX2GGTX3A (SEQ ID No: 1), in which each of X1 and X2 are independently G, C, or absent, and X3 is T or C, or L- (G)n-L', in which n is an integer of 5-9 inclusive, and L and L'are nucleotide segments having complementary sequences. Also provided herein are multimeric nucleic acid aptamers containing a backbone moiety, which comprises a palindromic sequence.

Description

[0001] related application [0002] This application claims the benefit under 35 U.S.C. §119 of U.S. Provisional Application No. 62 / 684,139, filed June 12, 2018, and U.S. Provisional Application No. 62 / 740,751, filed October 3, 2018, each of which is adopted in its entirety by Incorporated herein by reference. Background technique [0003] Activated T cells express a variety of co-inhibitory molecules (termed immune checkpoint molecules) to regulate T cell responses. Exemplary immune checkpoint molecules include programmed cell death protein 1 (PD-1), lymphocyte activation gene 3 (LAG-3), and cytotoxic T lymphocyte-associated protein 4 (CTLA-4). These immune checkpoint molecules play an important role in maintaining immune homeostasis and preventing autoimmunity. [0004] Immune checkpoint molecules are often activated in cancer, leading to suppressed anti-tumor immune responses. Therefore, immune checkpoint inhibitor therapy offers effective long-term treatment for a varie...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/115G01N33/68A61K31/7088A61P37/02A61P35/00
CPCC12N15/115A61P37/00A61P35/00C12N2310/16C12N2310/51
Inventor 张翼中C-H·蒋Y-W·高
Owner ONENESS BIOTECH
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