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Artificial single-chain deoxynucleotide having therapeutic effect to human B cell tumour

A deoxynucleotide and artificial synthesis technology, which is applied in the field of preparations for the treatment of human B lymphocyte tumors, can solve the problems of growth inhibition of B lymphocyte tumor cells, etc.

Active Publication Date: 2006-11-22
CHANGCHUN HUAPU BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0012] 2) The growth of B lymphocyte tumor cells is inhibited;

Method used

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  • Artificial single-chain deoxynucleotide having therapeutic effect to human B cell tumour
  • Artificial single-chain deoxynucleotide having therapeutic effect to human B cell tumour
  • Artificial single-chain deoxynucleotide having therapeutic effect to human B cell tumour

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Embodiment 1, the design of the artificially synthesized single-stranded deoxynucleotide

[0044] The design sequence is as follows:

[0045] 1. The artificially synthesized single-stranded deoxynucleotide (X1=A, T, G; X2=A, T; Y1=A, T; Y2=A, T, C; L, M=A, T, C, G; n is 0-6)

[0046] 5'-ggggTCgTTCgTCgTTgggggg-3' (SEQ ID NO: 1) [121]

[0047] 5'-ggggATAACgTTgCgggggg-3' (SEQ ID NO: 2) [143]

[0048] 5'-ggggTgCAACgTTCAgggggg-3' (SEQ ID NO: 3) [402]

[0049] 5'-ggggTCCTACgTAggAgggggg-3' (SEQ ID NO: 4) [123]

[0050] 5'-ggggTCCATgACgTTCCTgAAgggggg-3' (SEQ ID NO: 5) [603]

[0051] 5'-gggggACgTCgCCgggggggg-3' (SEQ ID NO: 6) [118]

[0052] 5'-ggATCCgTACgCATgggggg-3' (SEQ ID NO: 7) [320]

[0053] 5'-ggggggAATCgATTCgggggg-3' (SEQ ID NO: 8) [154]

[0054] 5'-gggATgCATCgATgCATCgggggg-3' (SEQ ID NO: 9) [464]

[0055] 5'-ggTgCgACgTCgCAgggggg-3' (SEQ ID NO: 10) [471]

[0056] 5'-gggACgTACgTCgggggg-3' (SEQ ID NO: 11) [390]

[0057] 5'-gggggATCgACgTCgATCgggggg-3' (SEQ ID NO: ...

Embodiment 2

[0235] Embodiment 2, the synthesis of single-stranded deoxynucleotide

[0236] The solid-phase phosphoramidite triester method is used to synthesize DNA fragments. This method has the advantages of high efficiency and rapidity, and has been widely used in DNA chemical synthesis.

[0237] DNA chemical synthesis is different from enzymatic DNA synthesis. The enzymatic DNA synthesis process extends from the 5'→3' direction, while DNA chemical synthesis starts from the 3' end. Concrete reaction steps are as follows:

[0238] 1. Deprotection group

[0239] Use trichloroacetic acid to remove the protective group DMT (dimethoxytrityl) of the nucleotide attached to CPG (Controlled Pore Glass) to obtain a free 5'-hydroxyl terminal for the next condensation reaction use.

[0240] 2. Activation

[0241] The phosphoramidite-protected nucleotide monomer is mixed with tetrazolium activator and enters the synthesis column to form the phosphoramidite tetrazolium active intermediate (its 3...

Embodiment 3

[0249] Example 3, Expression of Toll-like receptor 9 in human B lymphocytic leukemia cells

[0250] 1. Preparation of human chronic B lymphocytic leukemia cells

[0251] Peripheral blood mononuclear cells (PBMC) were obtained from the blood of human chronic B lymphocytic leukemia patients by Ficoll-hypaque gradient centrifugation. CD5 was isolated using B lymphocyte isolation kit (Miltenyi Biotec, Bergisch Gladbach, Germany) + CD19 + CD23 + Human chronic B lymphocytic leukemia cells with a purity of 95%. The specific operation was carried out according to the method provided by Miltenyi Biotec Company. Using the B or T lymphocyte isolation kit (MiltenyiBiotec, Bergisch Gladbach, Germany), the B or T lymphocytes isolated from normal human PBMC, the obtained B lymphocytes (CD19 + ) and the obtained T lymphocytes (CD3 + ) have a purity of more than 95%. The specific operation was carried out according to the method provided by Miltenyi Biotec (Bergisch Gladbach, Germany). ...

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PUM

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Abstract

The invention discloses an artificial single-chained deoxyriboside and B cell tumour treatment technology and method, which is characterized by the following: inducing B cell tumour occur dying; reinforcing B cell tumour cell to express CD40; treating B cell tumour.

Description

technical field [0001] The invention provides a preparation for treating human B lymphocyte tumors, in particular to artificially synthesized single-stranded deoxynucleotides, which can induce apoptosis of human B lymphocyte tumors and enhance the expression of CD40 in human B cell tumor cells , has a therapeutic effect on human B lymphocyte tumors. technical background: [0002] According to the World Health Organization classification system (WHO classification system), human lymphoid malignancies (lymphoid malignancies) are divided into three main types: B-cell neoplasms (B-cell neoplasms), T-lymphocyte / natural killer cell tumors (T-cell / natural killer(NK)-cell neoplasms) and Hodgkin's lymphoma. Human B lymphocyte tumors (B-cell neoplasms) are further divided into two categories: Precursor B-cell neoplasms and peripheral B-cell neoplasms. Precursor B-cell neoplasms include: precursor B-acute lymphoblastic leukemia / lymphoblastic lymphoma (B-ALL, LBL). Peripheral B-cell n...

Claims

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Application Information

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IPC IPC(8): C07H21/00A61K31/7088A61P35/00C12N15/117
CPCC12N15/117C12N2310/17C12N2310/313C12N2310/315A61P35/00A61P35/02A61P37/00A61P37/02A61P43/00C07H21/04C12N15/00A61K31/7088C12N15/11
Inventor 王丽颖包木胜于永利
Owner CHANGCHUN HUAPU BIOTECHNOLOGY CO LTD
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