Application of oridonin in medicine preparation

A technology of Rubescensin A and medicine, which is applied in its application field in the field of pharmacy, can solve the problems that there are no animal models or clinical experiment data of Rubescensin A recorded, and achieve good medicinal prospects and strong pharmacological effects Effect

Inactive Publication Date: 2007-07-11
RUIJIN HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In this patent application, it is only generally described that Rubescensin A can degrade the AML1-ETO fusion pro

Method used

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  • Application of oridonin in medicine preparation
  • Application of oridonin in medicine preparation
  • Application of oridonin in medicine preparation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] Oridonin A Induced Apoptosis of t(8;21) Leukemia Cells in Vitro

[0058] 1. Materials and methods

[0059] 1. Reagents and Instruments

[0060] The purity of oridonin is 98%-99.8%. The stock solution prepared by dissolving in DMSO (Sigma) to a concentration of 10-2 mol / L was stored at -20°C. Propidium iodide (PI) was a product of Sigma, USA. Propidium iodide was made into a 250 μg / mL stock solution with triple distilled water and stored at 4°C in the dark. Fetal bovine serum was purchased from Hyclone Company, USA. Annexin V detection kit (ApoAlert Annexin-V kit) was purchased from BD Biosciences, USA. The flow cytometer is a product of Beckman Coulter, USA. Fluorescence microscope (Olympus, BX60) and phase contrast microscope (Olympus, IMT) were purchased from Japan Olympus Company. Cytospin, a cell smear instrument, was purchased from Shandon Company, UK.

[0061] 2. Cell culture and treatment, cell morphology observation

[0062] Human t(8;21) leukemia cell l...

Embodiment 2

[0072] Molecular Mechanism of Oridonin A Inducing Kasumi-1 Cell Apoptosis

[0073] 1. Materials and methods

[0074] 1. Reagents and Instruments

[0075] For Rubescensine A, propidium iodide, and fetal bovine serum, see Example 1 "Materials and Methods". Rhodamine (Rhodamine, Rh123) is a product of Sigma Company in the United States. Rhodamine was made into a 10 μg / mL stock solution with triple-distilled water and stored at 4°C in the dark. Caspase-3 specific inhibitor Z-DQMD-FMK is a product of Sigma Company in the United States, dissolved in DMSO and stored at -20°C before use. Poly(ADP-ribose) polymerase (poly(ADP[adenosinediphosphate]-ribose) polymerase, PARP) antibody, anti-ETO antibody was purchased from Santa Cruz Biotech, USA, anti-Caspase-3, Caspase-9 antibody, horseradish peroxidized The HRP-labeled secondary antibody and immunoblotting chromogenic system were purchased from Cell Signaling, USA. Actin antibody was purchased from Sigma, USA, and Cytochromec Releas...

Embodiment 3

[0112] Study on the caspase-3 cleavage site of AML1-ETO fusion protein

[0113] 1. Materials and methods

[0114] 1. Reagents and Instruments

[0115] Oridonin, fetal bovine serum see Example 1 "Materials and Methods". Caspase-3 specific inhibitor Z-DQMD-FMK see Example 2 "Materials and Methods". Anti-ETO antibody was purchased from Santa Cruz Biotech, USA. Actin antibody was purchased from Sigma, USA. Site-directed mutagenesis kit QuickChange Site-Directed Mutagenesis Kit is a product of Stratagene, USA. The Plasmid Maxi Preparation Kit is a product of QIAGEN, Germany. The Gene Pulser electroporator and the Cuvette electroporation cup are products of Bio-Rad, USA.

[0116] 2. Caspase-3 specific inhibitor Z-DQMD-FMK inhibition experiment

[0117] Kasumi-1 cell culture is the same as in Example 1. During the experiment, the cells were treated with 2-4×10 5 / ml density inoculation culture, application of 2 μ M, 5 μ M two concentrations of Rubescensine A and in the presen...

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Abstract

The invention relates to an application of rabdosia element for preparing cute leukemia drug, and an application for preparing cell leukemia drug, wherein the rabdosia element can induce the death of leukemia cells externally; in body, it can prolong the service life of t (8; 21) leukemia mouse, or release the cancer growth internally; the rabdosia can activate the death channel, to degrade AML1-ETO protein which is key of leukemia, via activating caspase-3. The invention also finds one important caspase-3 cutting point.

Description

technical field [0001] The present invention relates to the use of oridonin, in particular to its application in the field of pharmacy. Background technique [0002] Myelogenous leukemia includes acute myelogenous leukemia (AML) and chronic myelogenous leukemia (CML). Acute myeloid leukemia (AML) is a heterogeneous hematologic malignancy and the most common type of acute leukemia occurring in adults. According to records, in 2003 alone, about 11,000 Americans were diagnosed with AML, and about 75% of them eventually died of the disease. In China, there are as many as 40,000 new patients with acute leukemia each year. AML patients manifest as normal differentiation of hematopoietic cells and blocked apoptosis pathway, abnormal proliferation of bone marrow hematopoietic progenitor cells, resulting in a large accumulation of immature blast cells in bone marrow and peripheral blood, affecting the production and function of normal blood cells, leading to bleeding and anemia in ...

Claims

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Application Information

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IPC IPC(8): A61K31/352A61P35/00
CPCA61K31/202A61K31/52A61K31/70A61P35/00A61P35/02
Inventor 陈竺周光飚陈赛娟王振义
Owner RUIJIN HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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