Application of compound Pinnatifolone A in the preparation of anti-Parkinson's drugs

Abstract

The invention belongs to the field of medical technology, and discloses the use of a compound Pinnatifolone A in the preparation of an anti-Parkinson's disease drug. The compound Pinnatifolone A is the main active ingredient of a traditional Chinese medicine lilac stem. The invention also discloses a preparation method of the compound Pinnatifolone A, wherein the lilac stem is extracted with alcohol, and then extracted with dichloromethane and ethyl acetate in sequence, and the dichloromethane extraction portion is separated by silica gel column chromatography, and then a mixed solvent of petroleum ether and acetone is used for recrystallization and purification, so as to obtain a target compound. In vivo tests show that the compound Pinnatifolone A of the invention has strong activity in anti-Parkinson's disease, clear efficacy, and small toxic and side effects, and can be used as a potential anti-Parkinson's disease drug.

Description

Technical Field

[0001] The invention belongs to the field of medical technology, and in particular relates to application of the compound Pinnatifolone A in preparing anti-Parkinson's drugs. Background Art

[0002] Parkinson's disease (PD) is a chronic, progressive neurodegenerative disease whose main pathological feature is the loss of dopaminergic neurons in the brain, leading to the appearance of motor and non-motor symptoms. The causes of this disease may involve genetic factors, exposure to environmental toxins, and aging of the nervous system. The motor symptoms of Parkinson's disease include resting tremor, muscle rigidity, bradykinesia, and postural balance disorders, while non-motor symptoms include depression, anxiety, dementia, decreased or absent sense of smell, drooling, dysphagia, constipation, frequent and urgent urination, insomnia, excessive daytime sleepiness, fatigue, etc.

[0003] Modern medical treatments for Parkinson's disease mainly include drug therapy, surgical treatment, exercise therapy, psychological intervention, and care and nursing. Among them, drug therapy is the first choice and the main treatment method. Therapeutic drugs include compound levodopa, dopamine receptor agonists (DAs), monoamine oxidase type B inhibitors (MAO-BI), catechol-O-methyltransferase inhibitors (COMTI) and anticholinergics. However, existing treatments have limitations, such as drug side effects and complications caused by long-term use. For example, long-term use of levodopa may cause motor complications such as dyskinesia, and may also cause neuropsychiatric side effects such as hallucinations and impaired consciousness. In addition, some drugs may cause other side effects such as orthostatic hypotension and edema. Therefore, the development of drugs for the treatment of Parkinson's disease that are non-toxic, easy to take, and economical has become a hot spot in modern drug development.

[0004] Compound Pinnatifolone A (C 15 H 20 O3, molecular weight 248.141) is an erimofenane-type sesquiterpenoid compound with a wide range of pharmacological activities extracted and separated from plants of the genus Syringa in the family Oleaceae, and belongs to natural sesquiterpenoid compounds. Due to the novel structure of sesquiterpenoid compounds, modern research has found that they have a wide range of biological activities, including anti-tumor, antibacterial, anti-inflammatory, anti-neurotoxic, antiviral, immunosuppressive, hepatoprotective and cardiotonic effects. The latest research shows that Pinnatifolone A has the potential to be used as a sedative and hypnotic drug. At present, traditional Chinese medicine is a valuable resource for identifying and developing new drugs. Pinnatifolone A, as an active ingredient of traditional Chinese medicine, has the potential for further development. In addition, the preparation method of the compound Pinnatifolone A in the prior art is relatively complicated, which brings inconvenience to practical application. Summary of the invention

[0005] The purpose of the present invention is to explore the traditional Chinese medicine activity of compound Pinnatifolone A and provide the use of compound Pinnatifolone A in the preparation of anti-Parkinson's disease drugs. The inventors found in the study that the compound has strong activity in anti-Parkinson's disease, clear efficacy, and low toxic and side effects, and can be used as a potential anti-Parkinson's disease drug.

[0006] The technical solution of the present invention is as follows:

[0007] Application of the compound Pinnatifolone A in the preparation of anti-Parkinson's drugs.

[0008] Preferably, the chemical structural formula of the compound Pinnatifolone A is:

[0009] .

[0010] Preferably, the stereo configuration of the compound Pinnatifolone A is any one of the following:

[0011] .

[0012] More preferably, the stereo configuration of the compound Pinnatifolone A is .

[0013] Preferably, the content of the compound Pinnatifolone A in the drug is 1-99.5%, more preferably 2-50%.

[0014] Preferably, the drug further comprises a pharmaceutically acceptable carrier or excipient.

[0015] Those skilled in the art can directly or indirectly add the compound to various commonly used pharmaceutically acceptable carriers or excipients (such as fillers, disintegrants, lubricants, adhesives, etc.) required for preparing different dosage forms, and prepare commonly used oral preparations or injection preparations using conventional pharmaceutical preparation methods in the pharmaceutical field.

[0016] Preferably, the oral preparation is one of tablets, capsules, granules, fat emulsions, microcapsules, and pills.

[0017] Preferably, the injection preparation is an injection solution or a powder injection.

[0018] Preferably, the dosage of the oral preparation is 5-200 mg / kg of the compound Pinnatifolone A, more preferably 10-40 mg / kg.

[0019] The present invention also provides a method for preparing the compound Pinnatifolone A, comprising the following steps:

[0020] (1) Take the dried stems of Syringa japonica var. Syringa and add 10 times the amount of 70% (volume percentage) ethanol aqueous solution to extract twice, each time for 2 hours, combine the extracts, concentrate until there is no alcohol taste, extract with dichloromethane and ethyl acetate respectively, and obtain a dichloromethane extraction part and an ethyl acetate part;

[0021] (2) Take the dichloromethane extraction part and mix it with 100-200 mesh silica gel in a ratio of 1:1 and dry it. Then, separate it by silica gel (200-300 mesh) column chromatography, and use petroleum ether-acetone (volume ratio of 100:0, 50:1, 25:1, 10:1, 5:1, 1:1, 0:100) as the mobile phase for elution. Equal amounts are collected in units of 2000 ml, and combined and concentrated by thin layer chromatography to obtain 12 fractions 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12;

[0022] (3) Fraction 3 was recrystallized and purified in a mixed solvent of petroleum ether and acetone in a volume ratio of 5:1. Compound Pinnatifolone A was obtained after repeated recrystallization and purification.

[0023] The beneficial effects of the present invention are:

[0024] 1. The present invention uses the experiment of the effect of compound Pinnatifolone A on the climbing pole time and rotating rod fatigue exercise time of MTPT-induced mice to prove that the compound Pinnatifolone A in the present invention has strong activity in anti-Parkinson's disease, clear efficacy, and small toxic and side effects, and can be used as a potential anti-Parkinson's disease drug.

[0025] 2. The preparation method of the compound Pinnatifolone A of the present invention is simple, and the target compound can be obtained by silica gel column chromatography separation and recrystallization purification. Compared with the complicated preparation process in the prior art, it has more advantages and saves time and cost. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 This is the X-ray single crystal diffraction pattern of the compound Pinnatifolone A. DETAILED DESCRIPTION

[0027] The technical solutions in the embodiments of the present invention will be described clearly and completely below in conjunction with the accompanying drawings. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0028] Unless otherwise specified, the experimental methods involved in the embodiments of the present invention are all conventional methods, and the reagents involved are all commercially available products and can be obtained through commercial channels.

[0029] It should be noted that the erimofenane-type sesquiterpenoid compound Pinnatifolone A of the present invention can be extracted and separated from plants of the genus Syringa of the family Oleaceae, or can be prepared by other existing technologies.

[0030] The present invention provides another method for extracting and separating the compound Pinnatifolone A. Compared with the prior art that uses multiple separation steps to obtain the target compound Pinnatifolone A from lilac roots, the present invention extracts effective ingredients from lilac stems and only requires one silica gel column chromatography separation combined with recrystallization to obtain the target compound Pinnatifolone A. The method is simpler and saves time and cost.

[0031] 1. Preparation of compound Pinnatifolone A

[0032] The compound Pinnatifolone A of the present invention is prepared by this laboratory, extracted and separated from the plant Syringa oblata Lindl. of the genus Syringa in the family Oleaceae, and the specific preparation method is as follows:

[0033] The dried stems of Syringa japonica (10 kg) were crushed, soaked at room temperature for 24 hours, and extracted twice with 10 times the amount of 70% (volume percentage) ethanol aqueous solution, each for 2 hours, and the extracts were combined and concentrated until there was no alcohol taste, and extracted with dichloromethane (10 L) and ethyl acetate (10 L) respectively, to obtain 385 g of dichloromethane extraction and 296 g of ethyl acetate extraction. The dichloromethane extraction part was mixed with 100 mesh silica gel in a 1:1 ratio and dried, and then separated by silica gel (200-300 mesh) column chromatography, using petroleum ether-acetone (volume ratio 100:0, 50:1, 25:1, 10:1, 5:1, 1:1, 0:100) as the mobile phase, and equal amounts were collected in units of 2000 ml, and combined and concentrated by thin layer chromatography to obtain 12 fractions (1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12). Fraction 3 was recrystallized and purified in a mixed solvent of petroleum ether and acetone in a volume ratio of 5:1. After repeated recrystallization and purification, compound Pinnatifolone A (10.2 g) was obtained. The purity was more than 95% as detected by high performance liquid chromatography area normalization method. The structure of the compound was identified by mass spectrometry and nuclear magnetic resonance (1H-NMR, 13C-NMR, 2D-NMR). After structural analysis and comparison with literature, the compound was determined to be Pinnatifolone A with a molecular formula of C 15 H 20 O3, molecular weight is 248.14, and its planar chemical structure is as follows:

[0034] .

[0035] Also through Figure 1 The X-ray single crystal diffraction pattern analysis can determine the stereo configuration of the compound Pinnatifolone A as follows:

[0036] .

[0037] 2. Experimental study on the efficacy of compounds

[0038] 1. Experimental Animals

[0039] C57BL / 6J mice, 8–10 weeks old, male, weighing 22 ± 2 g, were purchased from Hunan Slake Jingda Experimental Animal Co., Ltd., license number SCXK (Xiang) 2024-000.

[0040] 2. Experimental methods

[0041] After one week of adaptive feeding, the mice were randomly divided into normal group, model group, low-dose Pinnatifolone A group (10 mg / kg), medium-dose Pinnatifolone A group (20 mg / kg), high-dose Pinnatifolone A group (40 mg / kg), and positive drug group (safinamide intragastrically, 40 mg / kg), with 9 mice in each group. The normal group was intragastrically administered with an equal volume of 0.5% CMC-Na (sodium carboxymethyl cellulose) solution, with a dosing volume of 0.1 ml / 10 g. After continuous administration or 0.5% CMC-Na for 9 days, starting from the 10th day, the rest of the groups except the normal group were intraperitoneally injected with MPTP (1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine, 25 mg / kg) and modeled at the same time. The drug was administered again 1 hour later for 5 consecutive days, a total of 14 days, and the rotarod and climbing pole behavior tests were performed on the 14th day.

[0042] 3. Experimental results

[0043] 1. Effect of compound Pinnatifolone A on the duration of pole climbing induced by MTPT in mice

[0044] The results are shown in Table 1. In the pole climbing experiment, normal mice were able to climb smoothly from the top of the pole to the bottom of the pole. Compared with the normal group, the movement time of the mice in the model group from the top of the pole to the bottom of the pole was significantly prolonged to 80.97±52.17 s. Due to the decreased mobility and muscle tremor in the model group, the mice were stationary on the pole, and the climbing process slowed down. Compared with the model group, the movement time of the mice on the pole was significantly shortened after administration of Pinnatifolone A at different doses.

[0045] Table 1

[0046]

[0047] 2. Effect of compound Pinnatifolone A on the duration of rotarod fatigue exercise in MTPT-induced mice

[0048] The rotarod fatigue exercise duration experiment showed that after MPTP injection, mice showed shortness of breath, tremor, tail stiffness, unstable gait, and limited behavioral ability. The results of the rotarod experiment are shown in Table 2. Compared with the control group, the exercise duration of mice in the model group on the rotarod fatigue instrument was significantly shortened to 11.18±8.37s. Compared with the model group, the exercise duration of mice in the rotarod fatigue instrument after administration of different doses of Pinnatifolone A was significantly prolonged.

[0049] Table 2

[0050]

[0051] In summary, it is proved that compound Pinnatifolone A has obvious anti-Parkinson's effect. Therefore, compound Pinnatifolone A can be used to prepare anti-Parkinson's drug.

[0052] Finally, it should be noted that the above is only a preferred embodiment of the present invention and is not intended to limit the present invention. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art can still modify the technical solutions described in the aforementioned embodiments or replace some of the technical features therein by equivalents. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. Use of the compound Pinnatifolone A in the preparation of anti-Parkinson's drugs, wherein the chemical structural formula of the compound Pinnatifolone A is: 。 2. The use according to claim 1, characterized in that: The content of the compound Pinnatifolone A in the medicine is 1-99.5%.

3. The use according to claim 1, characterized in that: The medicine also includes a pharmaceutically acceptable carrier or excipient.

4. The use according to claim 1, characterized in that: The dosage form of the drug is an oral preparation or an injection preparation.

5. The use according to claim 2, characterized in that: The content of the compound Pinnatifolone A in the medicine is 2-50%.

6. The use according to claim 4, characterized in that: The dosage of the oral preparation is 5-200 mg / kg of the compound Pinnatifolone A.

7. The use according to claim 6, characterized in that: The dosage of the oral preparation is 10-40 mg / kg of the compound Pinnatifolone A.

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