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A kind of pharmaceutical composition comprising c21 steroid saponin and application thereof

A technology of steroidal saponins and compositions, applied in the field of medicine, to achieve the effects of reducing severe side effects, high safety, and increasing clearance rate

Active Publication Date: 2022-07-22
JINAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no report on the neuroprotective activity and anti-β-amyloid activity of C21 steroidal saponins

Method used

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  • A kind of pharmaceutical composition comprising c21 steroid saponin and application thereof
  • A kind of pharmaceutical composition comprising c21 steroid saponin and application thereof
  • A kind of pharmaceutical composition comprising c21 steroid saponin and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0068] Example 1 Extraction and separation of C21 steroidal saponins

[0069] (1) Take 15kg of dried Radix Ginseng root powder and add 95% EtOH-H under refluxing conditions at 90°C. 2 O (4×30 L) was extracted for 2h to obtain 2.25kg of crude extract;

[0070] (2) Dissolve the crude extract in MeOH / H containing 5% HCl 2 O (2:1, 30 L) solution was refluxed for 3 h, and the pH of the reaction solution was slowly adjusted to 7.0 with 10% NaOH solution; then the organic solvent was removed in vacuo, and the residue was dissolved in H 2 O and EtOAc (3 x 30 L) to give a fraction soluble in EtOAc (1.48 kg);

[0071] (3) The EtOAc-soluble fraction obtained in step (2) was subjected to silica gel column chromatography (CC) with CH 2 Cl 2 -MeOH (gradient 95:1→80:1→60:1→30:1→15:1→10:1→5:1→2:1, v / v, 5 column volumes per gradient) Elution gave compound 1-5.

[0072] use 1 H NMR and / or 13 Compounds 1-5 were identified by C NMR, and the results are shown below.

[0073] Compound 1: ...

Embodiment 2

[0078] Example 2 C21 steroidal saponins promote the proliferation of hippocampal neurons

[0079] (1) Take HT22 cells in the logarithmic growth phase, and seed the cells in a 96-well plate at a density of 5,000 cells / well, with 3 replicate wells in each group;

[0080] (2) After culturing for 24 hours, different concentrations of compounds were added to continue the culture for 48 hours; the groups 1 to 6 were respectively added with compound 3 (20-O-VK) at concentrations of 0.01 μM, 0.1 μM, 0.5 μM, 1 μM, 5 μM, and 10 μM; Group C was the control group, and an equal volume of culture medium was added;

[0081] (3) After 48 hours, the medium containing the compound was removed, and 10 μL of DMEM medium containing CCK-8 was added to each well;

[0082] (4) at 5%CO 2 , 37℃ incubator, incubated for 2h;

[0083] (5) Shake for 15 s at room temperature, and measure the absorbance (OD) of each well at 450 nm with a microplate reader.

[0084] Test results such as figure 1 shown. ...

Embodiment 3

[0085] Example 3 Neuroprotective effect of C21 steroidal saponins on glutamate-induced damage to hippocampal neurons

[0086] (1) Take HT22 cells in the logarithmic growth phase, and seed the cells in a 96-well plate at a density of 5,000 cells / well, with 3 replicate wells in each group;

[0087] (2) After culturing for 24 hours, different concentrations of compounds were added to continue the culture for 24 hours; among them, groups 1 to 5 were added with compound 3 at concentrations of 0.1 μM, 0.5 μM, 1 μM, 5 μM, and 10 μM, respectively; the NC group was the blank group, and an equal volume was added. medium; group C was the control group, and an equal volume of medium was added;

[0088] (3) After 24h, 10mM glutamic acid was added to group 1-group 5 and group C respectively, and the NC group did not do any treatment and continued to culture for 24h;

[0089] (4) After 24 hours, the medium containing the compound was removed, and DMEM medium containing 10 μL of CCK-8 was ad...

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Abstract

The present invention relates to a pharmaceutical composition for the treatment, prevention and / or treatment of neurodegenerative diseases. On the one hand, its active ingredient C21 steroidal saponin can significantly reduce the production of β-amyloid in N2a-APP695 cells and increase excessive Aβ amyloid. Protein clearance rate; on the other hand, it promotes the proliferation of neuronal cells, and has a protective effect on neurotransmitters such as glutamate-induced neuronal cytotoxicity caused by the excessive accumulation of Aβ amyloid, thereby comprehensively playing a role in the treatment of neurodegenerative diseases. effect. Therefore, it has a good application prospect in the preparation of Aβ amyloid formation inhibitors and neuroprotective agents, and drugs for preventing or treating neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, Huntington's disease and the like.

Description

technical field [0001] The invention belongs to the field of medicine, and relates to a pharmaceutical composition comprising C21 steroidal saponins and applications thereof, in particular to the use of a pharmaceutical composition comprising C21 steroidal saponins in the preparation of Aβ amyloid formation inhibitors and neuroprotective agents application. Background technique [0002] Nerve cell damage is one of the main causes of neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, and Huntington's disease. The death of nerve cells can lead to cognitive, learning and memory dysfunction in patients with neurodegenerative diseases. Oxidative stress caused by neurotoxic substances such as beta amyloid and neurotransmitters is considered to be the main cause of nerve cell death. Among them, glutamate is the main endogenous neurotransmitter in the central nervous system, but high concentrations of glutamate can cause neurofibrillary tangles and neuron...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K31/57A61K36/27A61P25/28A61P25/16A61P25/14A23L33/105A61K125/00
CPCA61K31/57A61K36/27A61P25/28A61P25/16A61P25/14A23L33/105A23V2002/00A23V2200/322A23V2250/21
Inventor 邱佐成陈家旭唐紫灵庞倩倩李小叁鄢黎
Owner JINAN UNIVERSITY
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