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Preparation method of granzyme B activator curcumenol

A technology of curcumenol and ethanol, applied in biochemical equipment and methods, organic chemical methods, separation/purification of carbonyl compounds, etc., can solve problems such as complicated method steps, dependence on repeated column chromatography, and inapplicability to large-scale preparation

Inactive Publication Date: 2018-09-28
王欢
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There is currently no prior art that epiprocurcumenol, curcurenol, picrolactone A, picrolactone F, and vichakulide A can be deposited into the expression of granzyme B in CIK cells
[0006] In addition, the current methods for preparing epiprocurcumenol, curcumenol, picrolactone A, picrolactone F, and wechakucurin A are complicated in steps, rely on repeated column chromatography, and are not suitable for large-scale production.

Method used

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  • Preparation method of granzyme B activator curcumenol
  • Preparation method of granzyme B activator curcumenol
  • Preparation method of granzyme B activator curcumenol

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0084] Example 1: Effects of Epiprocurcumenol, Curcumenol, Kukulide A, Kukulide F and Wichakucurin A on the Expression Level of Granzyme B in CIK Cells (Granzyme B Activator)

[0085] 1. Experimental materials

[0086] Lymphocyte separation medium was purchased from Jiahe Biotechnology.

[0087] Fetal bovine serum and GT-T551 medium were purchased from Gibco, USA.

[0088] CD3mAB was purchased from Beijing Tongliyuan Biotechnology Co., Ltd., and IL-2 was purchased from Jiangsu Jinsili Pharmaceutical Co., Ltd.

[0089] 2. Experimental method

[0090] 1. Isolation and induction culture of CIK cells

[0091] Take 100mL of peripheral anticoagulant blood from healthy volunteers, add lymphocyte separation medium, centrifuge at 1500rpm for 15min, absorb the mononuclear cell layer, wash with normal saline three times, and centrifuge at 1500rpm for 10min each time; 5×10 6 cells / mL plus GT-T551 medium containing 500 μg / L CD3mAB, 1000 U / mL IL-2 and 10% FBS to induce and culture into...

Embodiment 2

[0113] Embodiment 2: the culture method of CIK cell and tumor killing activity assay

[0114] 1. Experimental materials

[0115] Lymphocyte separation medium was purchased from Jiahe Biotechnology.

[0116] Fetal bovine serum and GT-T551 medium were purchased from Gibco, USA.

[0117] CD3mAB was purchased from Beijing Tongliyuan Biotechnology Co., Ltd., and IL-2 was purchased from Jiangsu Jinsili Pharmaceutical Co., Ltd.

[0118] 2. Experimental method

[0119] 1. Culture of CIK cells

[0120] Take 100mL of peripheral anticoagulant blood from healthy volunteers, add lymphocyte separation medium, centrifuge at 1500rpm for 15min, absorb the mononuclear cell layer, wash with normal saline 3 times, and centrifuge at 1500rpm for 10min each time; 5×10 6 cells / mL plus GT-T551 medium containing 500 μg / L CD3mAB, 1000 U / mL IL-2 and 10% FBS to induce and culture into CIK cells, half of the medium was changed every 2-3 days.

[0121] After 7 days of induction culture, adjust the cel...

Embodiment 3

[0136] Embodiment 3: Preparation of granzyme B activator epigenal curcumenol, curcumenol

[0137] 1. Instruments and materials

[0138] TBE-300A high-speed countercurrent chromatography was purchased from Shanghai Tongtian Biochemical Technology Co., Ltd., China.

[0139] ADS-17 macroporous adsorption resin was purchased from Anhui Sanxing Resin Technology Co., Ltd.

[0140] 2. Separation method and results

[0141] 1. Preparation of epiprotocurcumenol and crude curcumenol

[0142] After crushing the curcuma herb, pass it through a 40-mesh sieve, take 800g and use 4L ethanol solution with a concentration of 75% by volume to extract by microwave for 20min at 400W power, extract twice, combine the filtrate and concentrate it under reduced pressure until it has no alcohol smell, and put it on the sample On the ADS-17 macroporous adsorption resin column (75cm × 3.5cm, resin 450mL), first eluted with 45% ethanol of 5 times of column bed volume, and then eluted with 75% ethanol o...

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Abstract

The invention discloses a preparation method of a granzyme B activator curcumenol. It is found that eiprocurcumenol, curcumenol, physagulin A, physagulin F and withangulatin A can activate the expression of granzyme B in CIK cells and are effective granzyme B activators; the granzyme B activators of eiprocurcumenol, curcumenol, physagulin A, physagulin F and withangulatin A can significantly improve the killing capability of CIK cells on tumor cells. The invention further provides methods for preparing eiprocurcumenol, curcumenol, physagulin A, physagulin F and withangulatin A. The methods areindependent of repeated column chromatography and suitable for industrialized large-scale preparation. The technical scheme disclosed by the invention has not been disclosed in the prior art yet.

Description

technical field [0001] The invention belongs to the field of biology, and in particular relates to the cultivation of CIK cells and the preparation method of inducing compounds. Background technique [0002] Malignant tumors seriously endanger human health. Currently, conventional surgery, radiotherapy and chemotherapy cannot completely eliminate tumor cells in the body. Adoptive immunotherapy has become an important adjuvant therapy for tumors due to its advantages of conforming to physiology, low toxicity and high efficiency. Among them, cytokine-induced killer cells (cytokine-induced killer cells, CIK cells) are a very promising adoptive immune cell discovered in recent years, with the advantages of rapid proliferation, broad-spectrum and high-efficiency tumoricidal activity, and minimal toxic and side effects. It has become a new option in the field of tumor immunotherapy. CIK is a heterogeneous cell population dominated by CD3+CD56+T cells, which has the characteristic...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07D493/08C07C45/78C07C49/743C07J71/00C12N5/0783A61P35/00
CPCC07D493/08A61P35/00C07B2200/07C07C45/78C07C49/743C07C2602/26C07J71/001C12N5/0638C12N2500/30
Inventor 王欢
Owner 王欢
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