阅读说明：本技术 金发藓酮a在制备治疗阿尔兹海默症药物中的应用 (Application of trichophyton A in preparation of medicine for treating Alzheimer's disease ) 是由 郭龙 郑玉光 段绪红 张丹 侯芳洁 景松松 于 2021-08-10 设计创作，主要内容包括：本发明公开了金发藓酮A在制备治疗阿尔兹海默症药物中的应用。金发藓酮A为本发明发明人先前发现并发表公开的一个化合物,本发明研究发现该化合物具有明显的乙酰胆碱酯酶抑制活性,因此具备开发成治疗阿尔兹海默症的药物的前景。(The invention discloses application of trichophyton A in preparation of a medicine for treating Alzheimer's disease. The trichophyton A is a compound which is discovered and published by the inventor previously, and the research of the invention discovers that the compound has obvious acetylcholinesterase inhibition activity, so the trichophyton A has the prospect of being developed into a medicament for treating the Alzheimer disease.)

1. Application of trichophyton A in preparation of medicines for treating Alzheimer's disease.

2. A medicament for treating alzheimer's disease, characterized in that: the active component is trichophyton A.

3. The medicament for treating alzheimer's disease according to claim 2, wherein: also contains pharmaceutically acceptable auxiliary materials, and is prepared into pharmaceutically acceptable dosage forms.

4. The medicament for treating alzheimer's disease according to claim 3, wherein: the dosage forms comprise tablets, capsules, injections and oral liquids.

Technical Field

The invention belongs to the field of medicines, relates to new application of a known compound, and particularly relates to application of trichophyton A in preparation of a medicine for treating Alzheimer's disease.

Background

The Toyao-Chiense largeflower is a whole plant of Pogonatum inflexum (Lindb.) Lac of Chiyao-Chinesemedicine of Chiyama, grows on the humid soil wall of the forest edge, often grows in large pieces in the mountainous areas of south and north China, and has the effects of tranquilizing and allaying excitement, dissipating blood stasis and stopping bleeding. The inventor previously separated 1 new benzophenone compound from Tokya minor moss, the chemical structure of which is shown as follows, and named gold trichodermone A (reference: Tokya minor moss 1 new benzophenone compound, Chinese herbal medicine, Vol. 50, No. 6, 2019, 3 months).

The invention is especially proposed and created in order to develop and expand the medical application of the medicine.

Disclosure of Invention

The invention aims to provide application of trichophyton A in preparing a medicine for treating Alzheimer's disease.

The above purpose of the invention is realized by the following technical scheme:

application of trichophyton A in preparation of medicines for treating Alzheimer's disease.

A medicine for treating Alzheimer's disease contains trichophyton A as active ingredient.

Furthermore, the medicine also contains pharmaceutically acceptable auxiliary materials and is prepared into pharmaceutically acceptable dosage forms.

Further, the dosage forms include tablets, capsules, injections and oral liquids.

Has the advantages that:

the trichophyton A is a compound which is discovered and published by the inventor previously, and the research of the invention discovers that the compound has obvious acetylcholinesterase inhibition activity, so the trichophyton A has the prospect of being developed into a medicament for treating the Alzheimer disease.

Drawings

FIG. 1 is a graph showing the inhibition rate of both Fangchon A and Taclin, a positive drug, on acetylcholine esterase.

Detailed Description

The following examples are given to illustrate the essence of the present invention, but not to limit the scope of the present invention.

1. Experimental reagent and instrument

Acetylcholinesterase (AChE) was purchased from Sigma-Aldrich; 5,5' -dithiobis (2-nitrobenzoic acid) (DTNB), iodothioacetylcholine (ATCI), all available from Allantin reagents, Inc.; sodium Dodecyl Sulfate (SDS), dimethyl sulfoxide (DMSO), Tris-HCl (pH 8.0) were purchased from beijing solibao technologies ltd, and all other reagents were analytical grade. The trichophyton A is separated and prepared from Pogonatum inflexum (Lindb.) Lac of Dictamnus minutissima of Dictamaceae, and has purity of more than 98% determined by high performance liquid chromatography.

Victorivo plate reader (perkin elmer instruments ltd); XS205 DU model electronic analytical balance (Mettler-Torledo instruments, Inc.); high speed centrifuges (edn gmbh, germany); from Milli-Q ultrapure water systems (Merck, Germany).

2. Experimental methods

2.1 preparation of the solution

Preparing a Tris-HCl buffer solution: 5mL of 1M Tris-HCl was diluted with distilled water to 100mL and diluted to 50mM Tris-HCl buffer.

Preparation of acetylcholinesterase (AChE) solution: AChE was weighed as 1mg and dissolved in 10mL of Tris-HCl buffer to obtain a final acetylcholinesterase (AChE) solution of 0.1 mg/mL.

Preparation of iodothioacetylcholine (ATCI) solution: 1.10mg of ATCI was weighed and dissolved in 0.5mL of Tris-HCl buffer to prepare an ATCI stock solution with a concentration of 8 mM. 200 μ L of ATCI stock solution was diluted to 1mL with Tris-HCl buffer to give a final concentration of 1.6mM iodothioacetylcholine (ATCI).

Preparation of 5,5' -dithiobis (2-nitrobenzoic acid) (DTNB) solution: 2.30mg of DTNB was dissolved in 1mL of Tris-HCl buffer solution to prepare a DTNB stock solution with a concentration of 6mM, and 400. mu.L of DTNB stock solution was diluted to 1mL with Tris-HCl buffer solution to obtain a final 5,5' -dithiobis (2-nitrobenzoic acid) (DTNB) solution with a concentration of 2.4 mM.

Preparation of Sodium Dodecyl Sulfate (SDS) solution: 20mg of SDS was weighed and added to 5mL of Tris-HCl buffer solution to prepare a 0.4% final concentration Sodium Dodecyl Sulfate (SDS) solution.

Preparing a trichophyton A solution: weighing a proper amount of the trichophyton A, dissolving the trichophyton A in 1mL of DMSO to prepare a 4mM stock solution, and diluting 100 mu L of the stock solution with a Tris-HCl buffer solution to 5mL to prepare an 80 mu M trichophyton A solution. Taking out different volumes of the trichophyton A solution respectively, and diluting the trichophyton A solution into a series of suitable concentrations by using Tris-HCl buffer solution.

Preparing a tacrine solution: weighing 1.8mg of tacrine, dissolving the tacrine in 2mLDMSO to prepare 4mM stock solution, taking 10 mu L of the 4mM stock solution, adding 990 mu L of Tris-HCl buffer solution to prepare 40 mu M tacrine solution; appropriate amount of tacrine solution is respectively taken and diluted into a series of appropriate concentrations by using Tris-HCl buffer solution.

2.2 determination of acetylcholinesterase inhibitory Activity

The Ellman's method is adopted to determine the inhibitory activity of the trichophyton A on acetylcholinesterase. The experiment is respectively provided with a sample determination group, a sample control group, an enzyme solution control group, a blank control group and a positive control group (tacrine) sample group, the reaction is carried out in a 96-well plate, and each group of experiments is repeated for 3 times. Each well of the sample assay set was sequentially added with 30. mu.L (50mM) of Tris-HCl buffer, 10. mu.L (2.4mM) of DTNB solution, 10. mu.L (0.1mg/mL) of AChE solution, and 10. mu.L of sample solution at various concentrations, and incubated at 37 ℃ for 15 min. Then, 10. mu.L (1.6mM) of substrate ATCI solution was added, incubation was continued at 37 ℃ for 15min, and 30. mu.L (0.4%) of terminator SDS solution was rapidly added to terminate the reaction. Absorbance was measured at 405nm using a microplate reader.

The inhibition rate of different samples to acetylcholinesterase is calculated according to the following method, 5 different concentrations are respectively set for the trichophyton A and tacrine samples, the inhibition rate of the trichophyton A and tacrine with different concentrations to acetylcholinesterase is calculated, and the half Inhibition Concentration (IC) of the trichophyton A and tacrine to acetylcholinesterase inhibition is calculated₅₀)。

A_xiFor the sample assay set (10. mu.L enzyme solution + 10. mu.L sample);

A_xas a sample control (10. mu.L Tris-HCl buffer + 10. mu.L sample);

A_ienzyme solution control (10. mu. LTris-HCl buffer + 10. mu.L enzyme solution);

A₀blank control (20. mu.L Tris-HCl buffer).

3. Results of the experiment

The established method is used for determining the inhibition effect of the trichophyton A on acetylcholinesterase, and as can be seen from figure 1, the trichophyton A and a positive drug tacrine both show better acetylcholinesterase inhibition effect, and the inhibition rate is continuously increased along with the increase of the concentration, so that the trichophyton A and the positive drug tacrine are dose-dependent. Calculating to obtain half Inhibition Concentration (IC) of trichophyton A acetylcholinesterase₅₀) 1.583 + -0.03 μ M, half Inhibitory Concentration (IC) of tacrine as a positive drug₅₀) It was 0.910. + -. 0.02. mu.M. The result indicates that the trichophyton A shows good acetylcholinesterase inhibition capability which is similar to the inhibition capability of a positive drug tacrine.

In conclusion, the trichophyton A has obvious acetylcholinesterase inhibition activity, so the trichophyton A has a prospect of being developed into a medicament for treating the Alzheimer disease.

The above-described embodiments are intended to be illustrative of the nature of the invention, but those skilled in the art will recognize that the scope of the invention is not limited to the specific embodiments.