Traditional chinese medicine for preventing and / or treating parkinson's disease and method for preparing same

Abstract

Provided are a traditional Chinese medicine for preventing and / or treating Parkinson's disease and a method for preparing same. The traditional Chinese medicine comprises the following raw materials in parts by weight: 10-15 parts of Gastrodiae Rhizoma, 5-10 parts of Juglandis Semen, 30-40 parts of honey, 30 parts of Rosa roxburghii, and 5-8 parts of lemon. The traditional Chinese medicine has significant effects on the prevention and / or treatment of Parkinson's disease, and features simple composition, mild adverse effects, and good drug stability.

Description

Traditional Chinese medicines for the prevention and / or treatment of Parkinson's disease and their preparation methods Technical Field

[0001] This invention relates to a traditional Chinese medicine for the prevention and / or treatment of Parkinson's disease and its preparation method, belonging to the field of pharmaceutical technology. Background Technology

[0002] Parkinson's disease (PD) is the second leading cause of neurodegenerative disease worldwide and the fastest-growing neurological disorder. Its main pathological feature is the degeneration of dopaminergic neurons in the substantia nigra, leading to a deficiency of dopamine neurotransmitters in the striatum, which in turn causes an imbalance between dopamine and acetylcholine. Patients often present with motor symptoms such as tremor, rigidity, and bradykinesia, as well as non-motor symptoms such as sleep disturbances, sensory disturbances, autonomic dysfunction, and mental and cognitive impairments. Currently, there are over 6 million people with Parkinson's disease worldwide, and this number is projected to double by 2040.

[0003] Western medicine treatment for Parkinson's disease primarily alleviates symptoms by supplementing with levodopa, activating dopamine receptors, acting as anticholinergics, and inhibiting dopamine breakdown. However, these drugs have several problems: first, they cannot cure Parkinson's disease, only temporarily improving symptoms; second, long-term use can lead to side effects such as end-of-dose exacerbations, on-off phenomena, and dyskinesia, severely reducing patient adherence. Furthermore, Western medicine treatment is expensive, and its efficacy gradually decreases with prolonged use.

[0004] Traditional Chinese medicine (TCM) demonstrates comprehensive advantages in the treatment principles, drug compatibility, late-stage efficacy, and syndrome differentiation of Parkinson's disease, exhibiting stable and lasting therapeutic effects with minimal side effects. TCM considers the etiology and pathogenesis of Parkinson's disease to be complex, often related to liver and kidney deficiency, qi and blood insufficiency, and phlegm and blood stasis obstructing the meridians. TCM treatment emphasizes holistic regulation, effectively improving motor and non-motor symptoms with minimal side effects through methods such as tonifying the liver and kidneys, promoting blood circulation and removing blood stasis, and calming wind and stopping spasms. In recent years, numerous researchers have dedicated themselves to exploring the therapeutic effects of TCM compound formulas on Parkinson's disease, finding that TCM, used alone or in combination with Western medicine, can achieve good therapeutic effects.

[0005] Although some traditional Chinese medicine (TCM) compound prescriptions have been patented, existing research still has shortcomings. For example, most current TCM compound prescriptions target a specific symptom or stage, lacking a systemic treatment plan for the entire course of Parkinson's disease. Furthermore, the stability and bioavailability of most TCM drugs need improvement.

[0006] In conclusion, given the complex pathogenesis of Parkinson's disease, developing a traditional Chinese medicine composition with significant efficacy, minimal side effects, and broad applicability, along with its preparation method, is of great importance. This composition should combine modern medical theory with the principles of traditional Chinese medicine's syndrome differentiation and treatment, addressing multiple targets and levels to comprehensively regulate bodily functions. Simultaneously, the preparation process should be optimized to improve the drug's stability and efficacy.

[0007] Summary of the Invention

[0008] The first objective of this invention is to provide a traditional Chinese medicine for the prevention and / or treatment of Parkinson's disease.

[0009] To achieve the purpose of this invention, the traditional Chinese medicine for preventing and / or treating Parkinson's disease comprises, by weight, the following medicinal materials: 10-15 parts of Gastrodia elata, 5-10 parts of walnut, 30-40 parts of honey, 30 parts of prickly pear, and 5-8 parts of lemon.

[0010] In one specific embodiment, the traditional Chinese medicine comprises the following medicinal materials in parts by weight: 13-15 parts of Gastrodia elata, 7-10 parts of walnut, 33-35 parts of honey, 30 parts of prickly pear, and 5-8 parts of lemon.

[0011] In one specific embodiment, the traditional Chinese medicine comprises the following medicinal materials in parts by weight: 13 parts of Gastrodia elata, 10 parts of walnut, 35 parts of honey, 30 parts of prickly pear, and 5 parts of lemon.

[0012] In one specific embodiment, the preparation method of the traditional Chinese medicine includes: crushing the gastrodia elata, walnut, prickly pear, and lemon; mixing the crushed gastrodia elata, walnut, prickly pear, and lemon with honey and sealing them, and fermenting them for more than one year at 10-25°C under light-proof conditions.

[0013] A second objective of this invention is to provide a method for preparing the above-mentioned traditional Chinese medicine for the prevention and / or treatment of Parkinson's disease.

[0014] To achieve the second objective of this invention, the preparation method of the traditional Chinese medicine includes: pulverizing the gastrodia elata, walnut, prickly pear, and lemon; mixing the pulverized gastrodia elata, walnut, prickly pear, and lemon with honey and sealing them, and fermenting them for more than one year at 10-25°C under light-proof conditions. Beneficial effects

[0015] The traditional Chinese medicine of this invention has significant effects on the prevention and / or treatment of Parkinson's disease, and can improve the motor function of Parkinson's disease model animals and increase the levels of dopamine and its metabolites in the brain; the formula of this invention has simple ingredients, low toxicity, and good drug stability. Attached Figure Description

[0016] Figure 1. Schematic diagram of the mouse pole climbing test.

[0017] Figure 2 shows the comparison of pole climbing time for mice in each group. Among them, Control is the normal control group, MPTP is the model group, No.36 LD is the low-dose group of Jieben No.36 fermentation broth, and No.36HD is the high-dose group of Jieben No.36 fermentation broth.

[0018] Figure 3 shows a comparison of hind limb stride length in mice of different groups. Control is the normal control group, MPTP is the model group, No.36 LD is the low-dose group of Jieben No.36 fermentation broth, and No.36HD is the high-dose group of Jieben No.36 fermentation broth.

[0019] Figure 4 shows the striatal DA content of mice in each group, where Control is the normal control group, MPTP is the model group, No.36 LD is the low-dose group of Jieben No.36 fermentation broth, and No.36HD is the high-dose group of Jieben No.36 fermentation broth.

[0020] Figure 5 shows the DOPAC content of the metabolite in each group of mice. Among them, Control is the normal control group, MPTP is the model group, No.36 LD is the low-dose group of Jieben No.36 fermentation broth, and No.36HD is the high-dose group of Jieben No.36 fermentation broth.

[0021] Figure 6 shows the HVA content of the metabolite in mice in each group. Among them, Control is the normal control group, MPTP is the model group, No.36 LD is the low-dose group of Jieben No.36 fermentation broth, and No.36HD is the high-dose group of Jieben No.36 fermentation broth. Detailed Implementation

[0022] To achieve the purpose of this invention, the traditional Chinese medicine for preventing and / or treating Parkinson's disease comprises, by weight, the following medicinal materials: 10-15 parts of Gastrodia elata, 5-10 parts of walnut, 30-40 parts of honey, 30 parts of prickly pear, and 5-8 parts of lemon.

[0023] In one specific embodiment, the traditional Chinese medicine comprises the following medicinal materials in parts by weight: 13-15 parts of Gastrodia elata, 7-10 parts of walnut, 33-35 parts of honey, 30 parts of prickly pear, and 5-8 parts of lemon.

[0024] In one specific embodiment, the traditional Chinese medicine comprises the following medicinal materials in parts by weight: 13 parts of Gastrodia elata, 10 parts of walnut, 35 parts of honey, 30 parts of prickly pear, and 5 parts of lemon.

[0025] In one specific embodiment, the preparation method of the traditional Chinese medicine includes: crushing the gastrodia elata, walnut, prickly pear, and lemon; mixing the crushed gastrodia elata, walnut, prickly pear, and lemon with honey and sealing them, and fermenting them for more than one year at 10-25°C under light-proof conditions.

[0026] A second objective of this invention is to provide a method for preparing the above-mentioned traditional Chinese medicine for the prevention and / or treatment of Parkinson's disease.

[0027] To achieve the second objective of this invention, the preparation method of the traditional Chinese medicine includes: pulverizing the gastrodia elata, walnut, prickly pear, and lemon; mixing the pulverized gastrodia elata, walnut, prickly pear, and lemon with honey and sealing them, and fermenting them for more than one year at 10-25°C under light-proof conditions.

[0028] The specific embodiments of the present invention will be further described below with reference to examples, but the present invention is not limited to the scope of the embodiments described herein.

[0029] Example 1

[0030] Take 15 parts of Gastrodia elata, 7 parts of walnuts, 33 parts of honey, 30 parts of prickly pear, and 8 parts of lemon. Crush the Gastrodia elata, walnuts, prickly pear, and lemon; mix the crushed Gastrodia elata, walnuts, prickly pear, and lemon with honey and seal. Ferment for more than one year at 10-25℃ under light-proof conditions to obtain the traditional Chinese medicine Jieben No. 36 for the prevention and / or treatment of Parkinson's disease.

[0031] Example 2

[0032] Take 10 parts of Gastrodia elata, 8 parts of walnuts, 40 parts of honey, 30 parts of prickly pear, and 5 parts of lemon. Crush the Gastrodia elata, walnuts, prickly pear, and lemon into powder; mix the crushed Gastrodia elata, walnuts, prickly pear, and lemon with honey and seal. Ferment at 10-25℃ under light conditions for more than 1 year to obtain a traditional Chinese medicine for the prevention and / or treatment of Parkinson's disease.

[0033] Example 3

[0034] Take 15 parts of Gastrodia elata, 10 parts of walnuts, 30 parts of honey, 30 parts of prickly pear, and 8 parts of lemon. Grind the Gastrodia elata, walnuts, prickly pear, and lemon into powder; mix the powdered Gastrodia elata, walnuts, prickly pear, and lemon with honey and seal. Ferment at 10-25℃ under light conditions for more than 1 year to obtain a traditional Chinese medicine for the prevention and / or treatment of Parkinson's disease.

[0035] Example 4

[0036] Take 15 parts of Gastrodia elata, 5 parts of walnuts, 37 parts of honey, 30 parts of prickly pear, and 6 parts of lemon. Grind the Gastrodia elata, walnuts, prickly pear, and lemon into powder; mix the powdered Gastrodia elata, walnuts, prickly pear, and lemon with honey and seal. Ferment at 10-25℃ under light conditions for more than 1 year to obtain a traditional Chinese medicine for the prevention and / or treatment of Parkinson's disease.

[0037] The efficacy of Jieben No. 36 fermentation broth prepared in Example 1 was evaluated in an MPTP-induced C57BL / 6 mouse model of Parkinson's disease.

[0038] Experimental methods

[0039] A Parkinson's disease model was established in C57BL / 6 mice by inducing damage to dopamine neurons in the substantia nigra using MPTP, and the efficacy of Jieben 36 fermentation broth was evaluated. Animals were divided into four groups: a normal control group (Control, saline), a model group (MPTP + saline), a low-dose group (MPTP + 0.36 mL / d), and a high-dose group (MPTP + 0.72 mL / d) of Jieben 36 fermentation broth. MPTP was administered intraperitoneally at a dose of 30 mg / kg once daily for 5 days, followed by a 3-day interval to induce progressive damage to dopamine neurons. Starting on day 8, mice were administered the broth via gavage twice daily for 14 days. The main observation indicators were changes in body weight, mouse behavior (pole climbing, gait tests), striatal dopamine levels, and metabolites.

[0040] 1. Laboratory animals and their husbandry management

[0041] Species: Rodents

[0042] Department: C57BL / 6

[0043] Animal rating: SPF

[0044] Animal age: 10-12 weeks old

[0045] Sex: Male

[0046] Weight: 25±2 g

[0047] Number: 32 animals, including 8 normal control group, 8 model group, 8 low-dose group (0.36 mL / d) of Jieben No. 36 fermentation broth, and 8 high-dose group (0.72 mL / d) of Jieben No. 36 fermentation broth.

[0048] Animals were housed in accordance with the SPF laboratory animal husbandry and management procedures of Shenzhen Center for Disease Control and Prevention. The housing conditions were a constant natural light / dark cycle (12 h / 12 ​​h), a temperature of 23℃–25℃, and a humidity of 50%–65%. All procedures in the experiment were carried out in accordance with the regulations of the NIH and the laboratory animal ethics committee of Shenzhen Center for Disease Control and Prevention.

[0049] 2. Experimental Methods

[0050] 2.1. Animal Quarantine

[0051] After the C57BL / 6 mice arrive at the SPF-grade laboratory, they are strictly quarantined and inspected according to the relevant technical requirements of the SPF laboratory. The following conditions of the mice need to be closely observed and recorded:

[0052] General condition: Observe the mouse's fur luster, consciousness, mental state, gait, and response to stimuli to understand the animal's habits.

[0053] Movement: Observe the mice's walking ability and walking balance.

[0054] 2.2. Establishment of MPTP-induced subacute Parkinson's disease model in male C57BL / 6 mice

[0055] Animal preparation: Mice were trained to climb poles 3 times in advance, and mice with poor coordination were removed.

[0056] MPTP injection modeling: Animal weight was recorded, and male C57BL / 6 mice were intraperitoneally injected with MPTP at a rate of 0.1 mL / 10 g. Animals in the modeling group received intraperitoneal injections of MPTP·HCl (dissolved in physiological saline, MPTP·HCl 35.1 mg / kg, containing 30 mg / kg MPTP), while the control group received the same amount of physiological saline in the same manner. Injections were administered once daily for 5 consecutive days, followed by a 3-day break. On the third day after the MPTP administration ended, the MPTP-injected mice were randomly assigned to groups using a random number table for subsequent treatment with the fermentation broth from Jieben No. 36.

[0057] 2.3. Animal grouping and administration:

[0058] Table 1 Animal grouping

[0059]

[0060] The administration route was by gavage, starting on the third day after modeling, twice a day (at 9 am and 3 pm each day, 6 hours apart), for two consecutive weeks.

[0061] 2.4. Behavioral Indicators

[0062] 2.4.1. Pole test

[0063] A homemade pole, 50 cm long and 1 cm in diameter, was used. A small ball with a diameter of 1.5 cm was fixed to the top of the pole and wrapped with gauze to increase friction and prevent the mice from slipping. The pole was placed vertically in the animal cage, and the mice were placed head down on top of the pole. The time it took for the mice to climb from the top to the bottom platform was recorded. The mice were trained for three consecutive days before modeling, guided to climb down quickly, once a day. The value on the day before modeling was recorded as the normal value. The pole-climbing test was conducted on the 14th day after drug administration, using the time required for the mice to climb from the top to the bottom platform as the evaluation index. If the mice stopped or climbed in the opposite direction, the time was not recorded, and the measurement was repeated. For mice that could not hold the pole and slid down completely naturally, the climbing time was recorded as 60 seconds. The pole-climbing test pattern is shown in Figure 1.

[0064] 2.4.2. Gait analysis

[0065] The instrument used in the experiment was the CatWalkXT (brand: Noldus). Mice were trained for three consecutive days prior to modeling, guided to quickly traverse the walkway once daily. One day before the experiment, animals underwent necessary training to familiarize them with the gait walkway environment. During training, each time an animal could walk relatively steadily from one side of the walkway to the other without significant pauses or standing, the training was considered successful. Animals were placed on the left side, and the software automatically began data collection, stopping when the animal successfully reached the other side. The total distance traversed was analyzed, and the average stride length was calculated. The average stride length and the time required to traverse the walkway were used as evaluation indicators. If a mouse stopped midway or exhibited significant speed (step) inconsistencies, measurements were repeated, and the value from the day before modeling was recorded as the normal value. Gait testing was conducted on day 14 after drug administration.

[0066] 2.4.3. Electrochemical high-performance liquid chromatography (ECD-HPLC) system was used to detect changes in the contents of DA, DOPAC, and HVA.

[0067] At the end of the experiment, three mice from each group were randomly selected, anesthetized with 10% chloral hydrate, decapitated, and their striatum and substantia nigra were harvested and weighed on ice. The samples were then quickly placed on dry ice and stored at -80 °C for later use. 0.1 M perchloric acid was added at a v / w ratio (10 μL: 1 mg), homogenized on ice for 5 min, and then centrifuged at 12000 rpm for 10 min. 20 μL of the homogenate was injected for analysis. An Agilent Eclipse Plus C18 reversed-phase column (4.6 x 150 mm, 5 μm) was used. The mobile phase was methanol:water = 10:90 (containing 0.05 mol / L sodium dihydrogen phosphate, 0.03 mmol / L EDTA, and 0.8 mmol / L sodium octanesulfonate per liter of water). The flow rate was 1 mL / min, the column temperature was 33 °C, and the detection voltage was 0.52 V. The pH of the mobile phase was adjusted to 3.5. The contents of DA, DOPAC, and HVA are calculated from the standard curve based on the peak area.

[0068] The standard curves and quantitative calculation formulas for DA, DOPAC, and HVA are as follows:

[0069] DA:Y=7.6682X+0.9942 (R 2 =0.9999)

[0070] DOPAC: Y = 7.5563X + 1.8354(R) 2 =1)

[0071] HVA: Y = 1.4834X + 14.574 (R 2=0.9998)

[0072] 2.5. Statistical Analysis

[0073] Experimental data were analyzed using GraphPad Prism 5. Data are expressed as mean ± SD, and P < 0.05 was considered statistically significant.

[0074] 3. Experimental Results

[0075] 3.1. Behavioral performance of mice before and after the experiment

[0076] C57BL / 6 mice exhibited obvious tremors, piloerection, rapid breathing, tail hyperextension, arched back, reduced movement, and unsteady gait within 10-20 minutes after each intraperitoneal injection of MPTP. The piloerection and tremors disappeared within 24 hours, and the limb stiffness, reduced movement, and balance disorders also subsided within 24 hours. In contrast, mice in the saline group showed no obvious abnormalities.

[0077] 3.2. The effect of Jieben No. 36 fermentation broth on improving behavioral impairment in MPTP-induced PD mouse model.

[0078] Normal values ​​of pole climbing time in mice were recorded before MPTP administration (Day 1), and there were no significant differences among the groups (Table 2). Behavioral results 14 days after modeling (Day 22) showed that the pole climbing time in the model group was significantly longer than that in the normal control group, the pole climbing time in the low-dose group showed a decreasing trend, and the pole climbing time in the high-dose group was significantly reduced (see Figure 2).

[0079] Table 2. The reduction in pole-climbing time (seconds) in MPTP Parkinson's mice by the fermentation broth of Jieben No. 36.

[0080]

[0081] In Table 2 and Figure 2, data are presented as mean ± standard deviation. Each group has n = 8. Compared to the control group, ## p<0.01; compared with the model group, p<0.01.

[0082] 3.3. The fermentation broth of Jieben No. 36 can increase the stride length of MPTP mice.

[0083] Before MPTP administration (Day 1), the normal values ​​of mouse paw print stride length were analyzed, and there was no significant difference among the groups. After 14 days of drug administration following modeling (Day 22), the results showed that the hind limb stride length of the model group was significantly reduced. Different doses of Jieben No. 36 fermentation broth significantly increased the stride length of MPTP mice (see Figure 3 and Table 3).

[0084] Table 3. Fermentation broth No. 36 increased hind limb stride length in male MPTP Parkinson's mice.

[0085]

[0086] Data are presented as mean ± standard deviation. Each group has n = 8. Compared to the control group, ## p<0.01; compared with the model group, p<0.05 p<0.01.

[0087] 3.4. The fermentation broth of Jieben No. 36 significantly increased the content of DA and its metabolites in the striatum of MPTP-induced PD mouse models.

[0088] Compared with the normal control group, the striatal dopamine (DA) and its metabolites 3,4-dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA) in the model group were significantly reduced. Treatment with Jieben No. 36 fermentation broth significantly increased the striatal DA and its metabolites DOPAC and HVA (see Figures 4, 5, 6 and Table 4).

[0089] Table 4. The fermentation broth of Jieben No. 36 significantly increased the levels of Dopamine, DOPAC, and HVA in the striatum of male MPTP Parkinson's mice.

[0090]

[0091] Data are expressed as mean ± standard deviation. Each group has n=3. One-way ANOVA combined with the LSD test was used. Compared with the control group... ### p < 0.001, compared with the model group p<0.05 p<0.01.

[0092] 4. Experimental Conclusions

[0093] Different doses of Jieben 36 fermentation broth can improve the damage of MPTP to mouse behavior to a certain extent and significantly increase the content of DA and its metabolites DOPAC and HVA in the striatum of mice.

Claims

1. A traditional Chinese medicine for the prevention and / or treatment of Parkinson's disease, characterized in that, By weight, the Chinese medicine comprises the following medicinal materials in the following weight parts: 10-15 parts of Gastrodia elata, 5-10 parts of walnut, 30-40 parts of honey, 30 parts of prickly pear, and 5-8 parts of lemon.

2. The traditional Chinese medicine for the prevention and / or treatment of Parkinson's disease according to claim 1, characterized in that, The Chinese medicine comprises the following ingredients in parts by weight: 13-15 parts of Gastrodia elata, 7-10 parts of walnuts, 33-35 parts of honey, 30 parts of prickly pear, and 5-8 parts of lemon.

3. The traditional Chinese medicine for the prevention and / or treatment of Parkinson's disease according to claim 1, characterized in that, The Chinese medicine comprises the following ingredients by weight: 13 parts Gastrodia elata, 10 parts walnuts, 35 parts honey, 30 parts prickly pear, and 5 parts lemon.

4. The traditional Chinese medicine for the prevention and / or treatment of Parkinson's disease according to any one of claims 1 to 3, characterized in that, The preparation method of the traditional Chinese medicine includes: crushing the gastrodia elata, walnut, prickly pear, and lemon; mixing the crushed gastrodia elata, walnut, prickly pear, and lemon with honey and sealing them, and fermenting them for more than one year at 10-25°C under light-proof conditions.

5. The method for preparing the traditional Chinese medicine for the prevention and / or treatment of Parkinson's disease according to any one of claims 1 to 3, characterized in that, The preparation method includes: crushing the gastrodia elata, walnut, prickly pear, and lemon; mixing the crushed gastrodia elata, walnut, prickly pear, and lemon with honey and sealing them, then fermenting them at 10-25°C under light-proof conditions for more than one year.

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