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WO2009062374A1 - Utilisation pharmaceutique de liquiritigénine pour préparer un médicament destiné au traitement de maladies neurodégénératives - Google Patents

Utilisation pharmaceutique de liquiritigénine pour préparer un médicament destiné au traitement de maladies neurodégénératives Download PDF

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Publication number
WO2009062374A1
WO2009062374A1 PCT/CN2008/001705 CN2008001705W WO2009062374A1 WO 2009062374 A1 WO2009062374 A1 WO 2009062374A1 CN 2008001705 W CN2008001705 W CN 2008001705W WO 2009062374 A1 WO2009062374 A1 WO 2009062374A1
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WIPO (PCT)
Prior art keywords
glycyrrhizin
disease
group
mice
neurodegenerative disease
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PCT/CN2008/001705
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English (en)
Chinese (zh)
Inventor
Fugui Cui
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D311/00—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings
    • C07D311/02—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings ortho- or peri-condensed with carbocyclic rings or ring systems
    • C07D311/04—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring
    • C07D311/22—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4
    • C07D311/26—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4 with aromatic rings attached in position 2 or 3
    • C07D311/28—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4 with aromatic rings attached in position 2 or 3 with aromatic rings attached in position 2 only
    • C07D311/32—2,3-Dihydro derivatives, e.g. flavanones
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/14—Drugs for disorders of the nervous system for treating abnormal movements, e.g. chorea, dyskinesia
    • A61P25/16—Anti-Parkinson drugs
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia

Definitions

  • Glycyrrhizin is prepared for treatment
  • the present invention relates to the use of glycyrrhizin for the preparation of a medicament for the treatment of neurodegenerative diseases, and in particular to the use of glycyrrhizin for the preparation of a medicament for the treatment of senile dementia and Parkinson's disease.
  • Background technique
  • Glycyrrhiza is the root and rhizome of various plants of the genus Leguminosae (Glycyrrhiza Linn). It is listed in the "Shen Nong's Herbal Classic" and is listed as the top grade. It is beneficial to qi, clearing away heat and detoxifying, stopping bleeding, relieving pain, and relieving pain. Reconcile the effects of various drugs. In recent years, it has been found that licorice not only has strong detoxification effect, but also anti-ulcer, anti-inflammatory, antitussive and antitussive, lowering blood pressure, lowering blood fat and anti-cancer effects.
  • the chemical constituents of licorice are mainly flavonoids and triterpenoids, as well as a small amount of alkaloids, lignin and coumarin.
  • the dihydroflavonoid Liquir it in has a molecular formula of C 2i H 22 0 9 and a molecular weight of 418. 4.
  • the chemical name is 7-hydroxyxanthone-4, -0-glucoside (7- Hydroxyf lavanone 4' -0-glucos ide ).
  • the glycyrrhizin, or Liquir it igenin has a molecular formula of C 15 H 12 0 4 and a molecular weight of 256.3.
  • Alzheimer's disease is a chronic central nervous system degenerative disease with progressive cognitive decline as the main clinical symptom.
  • the incidence rate increases sharply with age.
  • the prevalence rate of the elderly over 65 years old in China is 1% ⁇ 5 %, and the age above 85 years old reaches 20% ⁇ 40%.
  • Pathological changes in Alzheimer's disease include extensive neuronal loss (mainly through apoptosis), numerous neuronal fiber tangles (neurof ibr i l lary tangles), and senile plaques (seni le plaque).
  • the currently marketed treatments for Alzheimer's disease include acetylcholinesterase inhibitors Arishin, Tacrine, etc., glutamate receptor blockers, memantine, etc. These drugs can improve or alleviate the symptoms of dementia in mild patients to some extent. , but can not prevent or reverse the further development of the lesion.
  • Parkinsons disease is also a common neurodegenerative disease.
  • the main pathological changes are degeneration and necrosis of dopaminergic neurons in the substantia nigra pars compacta, resulting in insufficient neurotransmitter dopamine, which ultimately leads to PD-specific Motor dysfunction.
  • levodopa remains the most effective drug for the treatment of Parkinson's disease.
  • such preparations have large side effects, and after several years of application, their efficacy is weakened, and switching phenomena and end-of-agent phenomena occur.
  • Recent experimental results suggest that levodopa may have a certain neurotoxic effect and accelerate the death of residual dopaminergic neurons.
  • the object of the present invention is to provide a novel use of glycyrrhizin, in particular for the use of glycyrrhizin for the preparation of novel medicaments for the treatment of neurodegenerative diseases, mainly Alzheimer's disease and Parkinson's disease.
  • glycyrrhizin has neuroprotective and nutritional effects in drugs selected for anti-neurodegenerative diseases, and no significant female activity (female activity refers to promotion of breast or endometrial epithelial cells). Proliferation, hyperplasia, etc., further studies have found that it promotes learning and memory in dementia model mice and enhances the ability of random movement in Parkinson's disease model mice. Therefore, glycyrrhizin can be used for the preparation of a medicament for preventing and treating senile dementia and Parkinson's disease.
  • glycyrrhizin pharmacological tests and results of glycyrrhizin will be used to illustrate their use in the preparation of novel drugs for the treatment of neurodegenerative diseases, mainly Alzheimer's disease and Parkinson's disease.
  • glycyrrhizin (1) Preparation of glycyrrhizin. After the commercially available glycyrrhizin was dissolved in methanol, an acetic acid buffer containing ⁇ -glucosidase (Sigma) was added. The ether was extracted three times and the ether layer was taken. The ether solution was then spun dry using a rotary evaporator, and the solid was scraped off and evaporated. The solid was dissolved in 10 to 20% acetonitrile, and the resolution was determined by high performance liquid chromatography (HPLC). Hirose determines the isolated compound.
  • HPLC high performance liquid chromatography
  • Hirose determines the isolated compound.
  • the glycyrrhizin of the present invention can be prepared into a dosage form such as a capsule, a tablet, a granule, a dropping pill and an injection after being added to an auxiliary material.
  • Glycyrrhizin has anti-neuronal apoptosis effect.
  • Flow cytometry was used to detect the apoptosis of rat pheochromocytoma cells (PC12) against glycyrrhizin against
  • the results showed that glycyrrhizin showed a significant anti-apoptotic effect at 0.2 g/ml.
  • Glycyrrhizin promotes the promotion of axonal growth in primary hippocampal neurons. Separation Primary rat hippocampal neurons were observed in vitro to promote the growth of neuronal axons by glycyrrhizin. The results showed that glycyrrhizin showed significant promotion of axonal growth at 0. 02 g/ml.
  • glycyrrhizin enhances memory.
  • the mouse jumping test and the Morri s water maze test were used to observe the promotion of learning and memory in scopolamine-induced Alzheimer's model mice.
  • the results showed that glycyrrhizin can significantly reduce the number of errors in the platform test, prolong the incubation period, shorten the latency of the Morr is water maze test, and increase the expression of cholinergic receptors in the brain of model mice, indicating that glycyrrhizin has enhanced learning and memory. effect.
  • glycyrrhizin has protective and nutritive effects on neurons, and can improve the learning and memory ability of the dementia model mice and the random exercise ability of the Parkinson's disease model mice without obvious female reproduction. Potential side effects of the system, etc.
  • the new drug prepared by the method provided by the invention has significant neuroprotective and neurotrophic effects without obvious female activity of flavonoids, and may have significant clinical effect without obvious toxicity, thereby generating great social benefits and economy. benefit.
  • detailed description Example 1 Preparation of glycyrrhizin
  • Example 5 Preparation of glycyrrhizin injection Take 30 g of glycyrrhizin raw material, 40 ml of propylene glycol, polyethylene glycol-400 100 ml, mix, heat in water bath for 30 min, add water for injection to 600 ml, treat in ultrasonic for 20 min, then heat in water bath to adjust pH 7.0-7.2. Filtration, potting, sterilization is available. Each 2ml, intramuscular injection, 1 time 2ml, 3 times a day, or mixed with normal saline, 5% glucose solution 500ml, intravenous drip, once a day, 3 times each time.
  • Example 6 Anti-apoptotic effect of glycyrrhizin
  • Flow cytometry was used to detect the apoptosis of rat pheochromocytoma cells (PC12) induced by glycyrrhizin against ⁇ 25 - 35 .
  • the experimental groups were as follows: control group; ⁇ 25 ⁇ 35 treatment group; ⁇ 25 - 35 different doses of drug group. Take 100 ⁇ of the cell suspension in a flow tube, add 5 ⁇ l Annexin V-FITC and ⁇ PI solution (2 (mg/ml), mix at room temperature for 15 minutes in the dark, add 400 ⁇ l PBS, and analyze by flow cytometry.
  • Rats were sterilized by volume fraction of 75% ethanol, and then decapitated in Pingya containing anatomical fluid. The whole brain was isolated and the hippocampus was separated under a dissecting microscope. It was placed in 0.25% trypsin and cut with scissors. Trypsin was pipetted into the serum containing tube every 4 min and fresh trypsin was added until no visible tissue pieces were visible. After filtering through a 200 mesh metal mesh, it was centrifuged at 1000 rpm ⁇ mirT 1 for 10 min.
  • the images were taken under a fluorescence microscope, and the length of the axons was counted by: 10 sheets per group, each measuring the length of axons of each neuron, and dividing by the total number of neurons after summation.
  • Glycyrrhizin itself has no promoting effect on the axon growth of primary hippocampal neurons, but
  • glycyrrhizin promoted axonal growth of primary hippocampal neurons in the concentration range of 0, 0 2 ⁇ 2 ⁇ ⁇ ⁇ mL- 1 (Table 2).
  • Table 2 Promoting effects of glycyrrhizin on axonal growth of primary hippocampal neurons Group concentration ( ⁇ g ⁇ mL - 1 ) Axon length ( ⁇ )
  • mice The promoting effect of glycyrrhizin on learning and memory in mice with Alzheimer's disease
  • the mouse jumping test and Morri s water maze test were used to observe the promoting effect of glycyrrhizin on learning and memory in scopolamine model mice induced by scopolamine.
  • Kunming mice weighing 14 ⁇ 16g, female. The mice were administered in groups on a random basis according to the principle of weight balance (see the table below for the dose). After 7 weeks of administration, the mold was molded and tested.
  • the mouse platform test face is as follows: The animal is placed on the platform, adapted to the environment for 3 min, and then subjected to 36 V voltage for 5 min. After 24 hours, the test recorded the latency of the first trip to the platform, the number of errors within 5 minutes, and the total shock time as a memory score.
  • the Morr i s water maze test was as follows: The mice were placed in a water maze one day before the test and free to swim for 2 min to suit the environment. Intraperitoneal injection of scopolamine 5 mg ⁇ kg 15 min before morning training. On the first day, the mice were trained 3 times. Each time, the mice were placed in the water from the 3 different water inlet points. If the mice could not find the underwater platform within 90s, they were placed directly on the platform. Stay for 30s. The next day, the training was conducted every day and afternoon. The time and path of the mice climbing the platform after entering the water were recorded as indicators for detecting the space and clues of the mice. After 24 hours, the test was repeated step by step. 3 days.
  • the results of the platform test showed that compared with the normal control group, the number of errors in the model group increased significantly within 5 minutes, and the latency was significantly shortened. Compared with the model group, the number of errors in the glycyrrhizin group was significantly reduced, and the incubation period was also significantly prolonged (see Table 3).
  • mice Male or female, weigh 18-22 grams.
  • CS-2 type mouse locomotor activity program instrument Development Institute of Materia Medica, Chinese Academy of Medical Sciences, 1-methyl-4-phenyl - 1, 2, 3, 6-tetrahydropyridine ⁇ set (1- methyl-4-phenyl -l, 2, 3, 6-tetrahydropyr idine, MPTP) is a product of Sigma.
  • the mice were randomly divided into four groups, 12 in each group, which were the normal control group, the model group, and the glycyrrhizin group (dose 30, 90 rag ⁇ kg -1 ).
  • mice After 0.54 hours of intraperitoneal injection of MPTP, except for the normal control group, the other groups of mice showed different degrees of dyskinesia, which showed slowness of movement, tremor, vertical hair, vertical tail and reduced activity. Compared with the model group, the number of autonomous activities in the glycyrrhizin group increased significantly during the test period, suggesting that glycyrrhizin can increase the central nervous system excitability and voluntary exercise ability of the model mice, and has a therapeutic effect on PD. The results are shown in Table 6. Effects of glycyrrhizin on the spontaneous activity of MPTP-induced Parkinson's disease model mice (5/5 min) Normal control 12 114. 8 ⁇ 26. 9 *** Model control 12 46. 1 ⁇ 27. 7
  • MCF-7 cells were passaged twice with phenol red-free 1640 medium containing 10% serum-free serum, digested by conventional methods, and seeded in 96-well plates at a concentration of 1.2 x 107180 ⁇ M/well. After 24 hours of cell adherence, add appropriate concentration of glycyrrhizin, glycyrrhizin or estradiol 20 ⁇ l/well, continue to culture for 72 hours, add MTT (5mg/ml) 20 ⁇ l / well before the end of the experiment, culture 4 Discard the supernatant after an hour, add DMS0 180 ⁇ ⁇ / hole, 0D value was measured at 540 nm after shaking and dissolved, and the cell proliferation rate was calculated. The results are shown in Table 7.
  • glycyrrhizin had no obvious cell proliferation at a concentration of 10 g/ml or less, and its glycoside glycoside significantly stimulated the proliferation of MCF-7 cells, indicating that glycyrrhizin had no obvious female activity, while glycyrrhizin had obvious effects.
  • the female activity suggests that the use of glycyrrhizin does not stimulate the potential side effects of proliferation or proliferation of the breast or endometrial epithelial cells. Effect of glycyrrhizin on proliferation of MCF-7 cells

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  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biomedical Technology (AREA)
  • Neurology (AREA)
  • Neurosurgery (AREA)
  • General Chemical & Material Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • Veterinary Medicine (AREA)
  • Medicinal Chemistry (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Psychology (AREA)
  • Hospice & Palliative Care (AREA)
  • Psychiatry (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

L'invention concerne l'utilisation pharmaceutique de liquiritigénine et d'une composition contenant de la liquiritigénine pour traiter des maladies neurodégénératives, en particulier la démence sénile ou la maladie de Parkinson.
PCT/CN2008/001705 2007-10-15 2008-10-08 Utilisation pharmaceutique de liquiritigénine pour préparer un médicament destiné au traitement de maladies neurodégénératives Ceased WO2009062374A1 (fr)

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CNA2007101635473A CN101152173A (zh) 2007-10-15 2007-10-15 甘草素在制备治疗神经退行性疾病药物中的用途
CN200710163547.3 2007-10-15

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WO2009062374A1 true WO2009062374A1 (fr) 2009-05-22

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Cited By (1)

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WO2021108855A1 (fr) * 2019-12-06 2021-06-10 Gretals Australia Pty Ltd Compositions de flavonoïde neurologiquements actives et leurs méthodes d'utilisation

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CN101152173A (zh) * 2007-10-15 2008-04-02 崔福贵 甘草素在制备治疗神经退行性疾病药物中的用途
CN101570528B (zh) * 2009-06-18 2011-09-28 吕秋军 甘草素衍生物及其制法和用途
CN103735497B (zh) * 2013-12-26 2016-04-20 南京海昌中药集团有限公司 甘草素注射液及其制备方法与应用
CN118759105B (zh) * 2024-07-11 2025-03-28 广州粤华制药有限公司 一种复方甘草片对ad治疗的药效质量研究方法

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WO2021108855A1 (fr) * 2019-12-06 2021-06-10 Gretals Australia Pty Ltd Compositions de flavonoïde neurologiquements actives et leurs méthodes d'utilisation

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