Use of traditional chinese medicine composition in preparation of pharmaceutical for preventing or / and treating amyotrophic lateral sclerosis

Abstract

The present invention belongs to the field of traditional Chinese medicine, and specifically relates to the use of a traditional Chinese medicine composition in the preparation of a pharmaceutical for preventing or / and treating amyotrophic lateral sclerosis. Specifically, the traditional Chinese medicine composition is prepared from herba schizonepetae, radix saposhnikoviae divaricatae, rhizoma seu radix notopterygii, radix angelicae pubescentis, radix bupleuri, radix peucedani, rhizoma chuanxiong, fructus aurantii, poria cocos, radix platycodi, and radix glycyrrhizae. Pharmacodynamic test results show that the traditional Chinese medicine composition provided by the present invention can prolong survival, maintain weight increase, and increase the limb strength of amyotrophic lateral sclerosis mouse models, has the effects of delaying the progression of muscle atrophy and delaying motor neuron apoptosis, and has a precise therapeutic effect for treating amyotrophic lateral sclerosis.

Description

Use of a traditional Chinese medicine composition in preparing a drug for preventing and / or treating amyotrophic lateral sclerosis Technical Field

[0001] The present invention belongs to the field of medical technology and relates to use of a traditional Chinese medicine composition in preparing a drug for preventing and / or treating amyotrophic lateral sclerosis. Background Art

[0002] Amyotrophic lateral sclerosis (ALS), also known as Lou Gehrig's disease, is a motor neuron disease characterized by involvement of the anterior horn cells of the spinal cord, motor nuclei in the brainstem, and the pyramidal tracts. It progresses rapidly and has a very poor prognosis, typically resulting in death from multi-organ failure 2 to 5 years after onset. It is a fatal neurodegenerative disease. Clinical manifestations include progressive skeletal muscle weakness, atrophy, fasciculations, bulbar palsy, and pyramidal tract signs. Some ALS patients may also experience varying degrees of cognitive and / or behavioral impairments, including frontotemporal lobe involvement. The disease generally develops in middle-aged and elderly individuals. In my country, the peak age of onset for ALS is around 50 years old, and the age of onset is trending younger, with a few patients presenting as young as 20 years old. Approximately 10% of ALS cases are familial, and multiple genes have been linked to the disease.

[0003] Currently, there is no definitive treatment for ALS. Only two US Food and Drug Administration (FDA)-approved drugs, riluzole and edaravone, are clinically used to potentially slow disease progression. In my country, potentially effective drugs such as butylphthalide, gangliosides, and mouse nerve growth factor are also commonly used. Some researchers have also tried using induced pluripotent stem cells and Vicocell (perampanel tablets), but none have demonstrated clear clinical efficacy. Traditional Chinese Medicine (TCM) uses syndrome differentiation and treatment based on the primary symptoms and other clinical features, combined with other treatments such as acupuncture, to slow disease progression.

[0004] Jingfang preparation is derived from Jingfang Baidu San (Jingfang Baidu San), a herb consisting of 11 Chinese herbs: Schizonepeta tenuifolia, Saposhnikovia divaricata, Notopterygium incisum, Angelica dahurica, Bupleurum chinense, Peucedanum chinense, Chuanxiong rhizome, Fructus Aurantii Immaturus, Poria cocos, Platycodon grandiflorum, and Licorice root. It has the effects of inducing diaphoresis, dispelling wind and dampness, and is used to treat colds, headaches, body aches, aversion to cold without sweating, nasal congestion, clear runny nose, and cough with white sputum. It has been clinically used to treat a variety of conditions, including acute viral upper respiratory tract infections, dengue fever, viral hepatitis, chickenpox, and mumps, with significant efficacy. However, there are no reports on the use of Jingfang preparation in the prevention and treatment of amyotrophic lateral sclerosis (ALS).

[0005] Summary of the Invention

[0006] The present invention aims to provide a use of a traditional Chinese medicine composition in preparing a medicament for preventing and / or treating amyotrophic lateral sclerosis.

[0007] Furthermore, the Chinese medicine composition is prepared from Schizonepeta tenuifolia, Saposhnikovia divaricata, Notopterygium wilfordii, Angelica dahurica, Bupleurum chinense, Peucedanum chinense, Chuanxiong, Fructus Aurantii Immaturus, Poria cocos, Platycodon grandiflorum, and Licorice.

[0008] Furthermore, the Chinese medicine composition is prepared from the following raw materials in parts by weight:

[0009] 30-100 parts of Schizonepeta, 30-100 parts of Saposhnikovia, 30-70 parts of Notopterygium, 30-50 parts of Angelica dahurica, 30-120 parts of Bupleurum, 30-60 parts of Peucedanum, 30-60 parts of Chuanxiong, 30-60 parts of Fructus Aurantii, 20-100 parts of Poria, 30-80 parts of Platycodon, and 10-25 parts of Licorice.

[0010] Furthermore, the Chinese medicine composition is prepared from the following raw materials in parts by weight:

[0011] 40-60 parts of Schizonepeta, 40-60 parts of Saposhnikovia, 40-50 parts of Notopterygium, 40-50 parts of Angelicae Pubescentis, 40-70 parts of Bupleurum, 40-50 parts of Peucedanum, 40-50 parts of Ligusticum chuanxiong, 40-50 parts of Fructus Aurantii Immaturus, 40-60 parts of Poria, 40-50 parts of Platycodon, and 12-18 parts of Radix Glycyrrhizae.

[0012] The traditional Chinese medicine composition can be added with other pharmaceutically acceptable excipients to prepare a clinically acceptable dosage form.

[0013] Furthermore, the clinically acceptable dosage forms are granules, oral solutions, mixtures, pills, syrups, capsules, and tablets.

[0014] The preparation method of the above-mentioned traditional Chinese medicine composition is an existing technology, including but not limited to the following methods: the preparation method published in the Ministry of Health Standard WS3-B-0328-90, the Ministry of Health Standard WS3-B-1377-93, Patent Application No. 202210119403.2, Patent Application No. PC T / CN2022 / 075573, etc.

[0015] Compared with the existing technology, the technical solution of the present invention has achieved the following beneficial technical effects: the results of pharmacodynamic experiments show that the traditional Chinese medicine composition of the present invention can improve the limb strength of mice with amyotrophic lateral sclerosis model, prolong their lifespan, maintain weight gain, and has the effect of delaying the progression of muscle atrophy and delaying the apoptosis of motor neurons, and can be used to treat amyotrophic lateral sclerosis. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 Effect of the Chinese medicine composition of the present invention on the survival rate of mice with amyotrophic lateral sclerosis

[0017] Figure 2 Effect of the Chinese herbal composition of the present invention on the body weight of mice with amyotrophic lateral sclerosis

[0018] Figure 3 Effect of the Chinese medicine composition of the present invention on the open field test in mice with amyotrophic lateral sclerosis

[0019] Figure 4 Effect of the Chinese herbal composition of the present invention on the fatigue rotarod test in mice with amyotrophic lateral sclerosis

[0020] Figure 5 Effect of the Chinese herbal composition of the present invention on the suspension experiment of amyotrophic lateral sclerosis mice

[0021] Figure 6 Effect of the Chinese herbal composition of the present invention on the grip strength test of mice with amyotrophic lateral sclerosis

[0022] Figure 7 Effect of Jingfang Granules on muscle atrophy in hSOD1 G93A mice DETAILED DESCRIPTION

[0023] In order to enable those skilled in the art to fully understand the present invention, the present invention is further described below through specific examples. However, those skilled in the art should be aware that the examples of the present invention do not limit the present invention in any way.

[0024] Example 1 Preparation of granules

[0025] prescription:

[0026] 75g of Schizonepeta tenuifolia, 75g of Saposhnikovia divaricata, 75g of Notopterygium wilfordii, 75g of Angelica dahurica, 75g of Bupleurum chinense, 75g of Peucedanum chinense, 75g of Chuanxiong, 75g of Citrus aurantium, 75g of Poria cocos, 75g of Platycodon grandiflorum, and 25g of Licorice root.

[0027] The preparation method refers to patent application number 202210119403.2.

[0028] Example 2 Granule Preparation

[0029] prescription:

[0030] 75g of Schizonepeta tenuifolia, 75g of Saposhnikovia divaricata, 65g of Notopterygium wilfordii, 35g of Angelica dahurica, 75g of Bupleurum chinense, 45g of Peucedanum chinense, 55g of Ligusticum chuanxiong, 45g of Citrus aurantium, 75g of Poria cocos, 75g of Platycodon grandiflorum, and 15g of Licorice root.

[0031] The preparation method refers to the Ministry of Health standard WS3-B-0328-90.

[0032] Example 3 Preparation of mixture

[0033] prescription:

[0034] 60g of Schizonepeta tenuifolia, 40g of Saposhnikovia divaricata, 50g of Notopterygium wilfordii, 40g of Angelica dahurica, 70g of Bupleurum chinense, 50g of Peucedanum chinense, 40g of Ligusticum chuanxiong, 50g of Citrus aurantium, 60g of Poria cocos, 40g of Platycodon grandiflorum, and 18g of Licorice root.

[0035] The preparation method refers to the Ministry of Health standard WS3-B-1377-93.

[0036] Example 4 Preparation of mixture

[0037] prescription:

[0038] 75g of Schizonepeta tenuifolia, 75g of Saposhnikovia divaricata, 75g of Notopterygium wilfordii, 75g of Angelica dahurica, 75g of Bupleurum chinense, 75g of Peucedanum chinense, 75g of Chuanxiong, 75g of Citrus aurantium, 75g of Poria cocos, 75g of Platycodon grandiflorum, and 25g of Licorice root.

[0039] The preparation method refers to patent application number PCT / CN2022 / 075573.

[0040] Example 5 Capsule Preparation

[0041] prescription:

[0042] Schizonepeta 40g, Saposhnikovia 60g, Notopterygium wilfordii 20g, Angelica pubescens 40g, Bupleurum 50g, Peucedanum 40g, Chuanxiong 50g, Citrus aurantium 40g, Poria 40g, Platycodon grandiflorum 50g, Licorice root 12g;

[0043] The preparation method refers to patent application number PCT / CN2022 / 075573.

[0044] Example 6 Effect of the Chinese medicine composition of the present invention on amyotrophic lateral sclerosis model mice

[0045] 1. Experimental Materials

[0046] Experimental animals: Male, 5-week-old amyotrophic lateral sclerosis model mice (hSOD1 G93A transgenic), weighing 21-23 g, were purchased from Jiangsu Jicui Yaokang Biotechnology Co., Ltd. Animal Use Permit Number: SYXK(Lu)2018-0008. Housing conditions were as follows: room temperature 20–26°C, with a diurnal temperature range ≤4°C; relative humidity 40%–70%; lighting: artificial lighting with a 12-hour light / dark cycle; air exchange rate ≥15 times / hour; and SPF rat growth and breeding diet with free access to water and food.

[0047] 18.1 Drugs:

[0048] Chinese herbal medicine extract: The clear paste prepared in Example 1 of the present invention and the volatile oil are mixed at a ratio of clear paste: volatile oil = 1 kg: 1.8 mL, and dissolved in an appropriate amount of water before use.

[0049] 2. Experimental methods and content

[0050] 2.1 Grouping and Dosing

[0051] All model mice were randomly divided into a model group and a drug administration group (aqueous solution of Chinese herbal extract, 2 g / kg), with 10 mice in each group. The drugs were administered by gavage every day for 28 consecutive weeks. The mice in the model group were gavaged with an equal dose of normal saline every day, and the gavage volume was 10 mL / kg.

[0052] 2.2 Observation indicators

[0053] 2.2.1 Body weight and survival rate

[0054] The body weight and growth status of the experimental mice in each group were monitored regularly every week.

[0055] 2.2.2 Open field experiment

[0056] The experiment was conducted in an open-field behavioral testing chamber. Mice were placed in the center square of the chamber's floor. A camera was fixed in the center of the chamber's top to record and time the movement of each animal. Each measurement lasted 5 minutes. The primary outcome measure was the distance traveled by the mouse within the open-field chamber. After the experiment, urine and feces were cleaned from each animal, and any remaining odor was removed with a small amount of 75% ethanol before the next animal could be tested.

[0057] 2.2.3 Grip test

[0058] First, a 3-day grip strength test was performed to adapt the mouse. During the formal test, the mouse was allowed to hold the grip bar with its two front paws. The tester held the mouse's tail to keep the mouse's body parallel to the grip strength meter, and ensured that the mouse's two front paws could exert force continuously and stably. The average grip strength during the continuous force exertion stage was recorded.

[0059] 2.2.4 Fatigue Rotarod

[0060] Place the mouse on the rotarod and record the time it remains on the rotarod after it begins to crawl. Each mouse is measured twice, with at least 1 hour between measurements.

[0061] 2.2.5 Suspension test

[0062] First, place the mouse on the lid of the cage and gently shake the cage lid to allow the mouse to grasp it. Then, invert the cage lid and record the time it takes for the mouse to fall from the inverted cage lid. Each mouse should be measured twice, with at least 1 hour between measurements.

[0063] 2.2.6 Statistical analysis

[0064] All data are expressed as mean ± SD. SPSS 19.0 statistical software was used for data analysis. One-way analysis of variance was used for comparisons between more than two groups, and T-test was used for comparisons between two groups. P < 0.05 was considered statistically significant.

[0065] 3. Experimental results and analysis

[0066] 3.1 Effect of the Chinese medicine composition of the present invention on the body weight and survival rate of model mice

[0067] During the experiment, the weight of mice in the drug-treated group showed a steady upward trend, while the weight gain of mice in the model group was smaller, with a significant difference compared to the drug-treated group (p < 0.01). At 24 weeks, the survival rate of mice in the model group was significantly decreased compared to the drug-treated group (p < 0.01). It is preliminarily speculated that the Chinese herbal composition of the present invention has a significant effect on improving the survival rate of ALS. The results are shown in Figures 1 and 2.

[0068] 3.2 Effects of the Chinese herbal composition of the present invention on the behavior of model mice

[0069] At 19 weeks, there was no significant difference in the behavioral indicators between the model group and the drug group (p>0.05). At 24 weeks, the grasping force, falling time in the hanging test, falling time in the fatigue rotating rod test, and movement distance of the model group were significantly shortened compared with the drug group (p<0.01), indicating that the Chinese medicine composition of the present invention has an effect on the behavioral manifestations of amyotrophic lateral sclerosis.

[0070] Significant improvement, the results are shown in Figures 3 to 6.

[0071] Example 7 Effect of the Chinese medicine composition of the present invention on amyotrophic lateral sclerosis model mice

[0072] 1 Experimental animals

[0073] SPF hSOD1 G93A male mice, 6 weeks old, weighing 21–23 g, were purchased from Jicui Yaokang Biotechnology Co., Ltd. Mice were housed in Laboratory 166, Barrier Area, Good Laboratory Practice (GLP), New Drug Pharmacology Center, Lunan Pharmaceutical Group Co., Ltd., under the following conditions: room temperature of 20–26°C, with a daily temperature range of ≤4°C; relative humidity of 40%–70%; artificial lighting with a 12-h / 12-h light / dark cycle; and ventilation rate of ≥15 times / hour. Mice were fed an SPF mouse and rat growth and breeding diet with free access to water and food.

[0074] 2 Methods

[0075] 2.1 Amyotrophic lateral sclerosis model

[0076] A human SOD1 fragment was obtained from a BAC containing the human SOD1 gene, and the amino acid at position 93 was mutated to construct a transgenic mouse model of hSOD1 G93A (glycine to alanine mutation at position 93). This model can be used for drug screening, safety evaluation, and pathogenesis research in ALS.

[0077] 2.2 Dosage regimen

[0078] All model group mice were randomly divided into model group (V), Jingfang group (4 g / kg), and riluzole group (Y: 15.7 mg / kg), with 10 mice in each group. The drugs were administered by gavage every day for 28 consecutive weeks. The mice in the model group were gavaged with an equal dose of injection water every day, and the gavage volume was 10 mL / kg.

[0079] 2.3 Grip test

[0080] First, a 3-day grip strength test was performed to adapt the mouse. During the formal test, the mouse was allowed to hold the grip bar with its two front paws. The tester held the mouse's tail to keep the mouse's body parallel to the grip strength meter, and ensured that the mouse's two front paws could exert force continuously and stably. The average grip strength during the continuous force exertion stage was recorded.

[0081] 2.4 Fatigue test

[0082] Place the mouse on the rotarod and record the time it remains on the rotarod after it begins to crawl. Each mouse is measured twice, with at least 1 hour between measurements.

[0083] 2.5 Muscle atrophy detection

[0084] After blood was collected from mice in each group under anesthesia, tissues from their left legs were obtained and the width of their gastrocnemius muscles was measured using a vernier caliper.

[0085] 2.6 Statistical analysis

[0086] All data are expressed as mean ± SD. SPSS 19.0 statistical software was used for data analysis. One-way analysis of variance was used for comparisons between more than two groups, and T-test was used for comparisons between two groups. P < 0.05 was considered statistically significant.

[0087] 3 Experimental results

[0088] 3.1 Effects of Jingfang Granules on Behavior (Tension) of hSOD1 G93A Mice

[0089] The experimental results are shown in Table 1. At 20-22W, there was no significant difference in the pulling force of mice in each group (p>0.05). At 24-28W, compared with the normal group, the pulling force of mice in the model group was significantly decreased (p<0.05). Compared with the model group, the pulling force of mice in the Jingfang group was significantly improved (p<0.05). The results are shown in Table 1.

[0090] Table 1 Effects of Jingfang Granules on the Tensile Strength of hSOD1G93A Mice (Statistical Table)

[0091] Table 1 Effect of Jingfang Granules on tension ofhSOD1 G93A mice(Statistical table)

[0092] Note: * indicates a difference compared with the normal group (P < 0.05); # indicates a difference compared with the model group (P < 0.05)

[0093] 3.2 Effects of Jingfang Granules on Behavior (Fatigue) of hSOD1 G93A Mice

[0094] The experimental results are shown in Table 2. At 28 weeks, compared with the normal group, the time for mice in the hSOD1 G93A model group to fall on the fatigue rotarod was significantly shortened (p<0.05). Compared with the hSOD1 G93A model group, the time for mice in the Jingfang group to fall on the fatigue rotarod was significantly improved. Due to the large individual differences, the standard deviation of the overall data was large, so no statistical difference was detected. The experimental results are shown in Figure 2.

[0095] Table 2 Effects of Jingfang Granules on Fatigue in hSOD1 G93A Mice (Statistical Table)

[0096] Table 2 Effect of Jingfang Granule on fatigue of hSOD1 G93A mice(Statistical table)

[0097] Note: * indicates a difference compared with the normal group (P<0.05)

[0098] 3.3 Effect of Jingfang Granules on Muscle Atrophy in hSOD1 G93A Mice

[0099] The experimental results are shown in Table 3. At 24 weeks, there was no significant change in the muscles of the mice in each group. At 28 to 30 weeks, compared with the normal group, the mice in the model group showed obvious muscle atrophy (p < 0.05). Compared with the model group, at 28 to 30 weeks, the muscle atrophy of the mice in the Jingfang group was significantly improved (p < 0.05). The experimental results are shown in Table 3.

[0100] Table 3 Effects of Jingfang Granules on Muscle Atrophy in hSOD1 G93A Mice (Statistical Table)

[0101] Table 3 Effect of Jingfang Granules on muscle atrophy in hSOD1 G93A mice(Statistical table)

[0102] Note: * indicates a difference compared with the normal group (P < 0.05); # indicates a difference compared with the model group (P < 0.05)

Claims

1. Use of a Chinese medicine composition in preparing a drug for preventing and / or treating amyotrophic lateral sclerosis, characterized in that: The composition comprises the following medicinal ingredients in parts by weight: 30-100 parts of Schizonepeta, 30-100 parts of Saposhnikovia, 30-70 parts of Notopterygium, 30-50 parts of Angelica dahurica, 30-120 parts of Bupleurum, 30-60 parts of Peucedanum, 30-60 parts of Chuanxiong, 30-60 parts of Citrus aurantium, 20-100 parts of Poria, 30-80 parts of Platycodon, and 10-25 parts of Licorice.

2. The use according to claim 1, characterized in that The prevention and / or treatment of ALS is manifested in improving the survival rate and / or limb strength of ALS patients.

3. The use according to claim 1, characterized in that The prevention and / or treatment of amyotrophic lateral sclerosis is manifested in delaying the apoptosis of motor neurons in patients with amyotrophic lateral sclerosis.

4. The use according to claim 1, characterized in that The prevention and / or treatment of amyotrophic lateral sclerosis is manifested in increasing the body weight of patients with amyotrophic lateral sclerosis.

5. The use according to claim 1, characterized in that The prevention and / or treatment of ALS is manifested in delaying the progression of muscle atrophy in ALS patients.

6. The use according to claim 1, characterized in that The composition is prepared from the various medicinal flavors.

7. The use according to claim 1, characterized in that The composition is prepared from the various medicinal flavors and pharmaceutically acceptable auxiliary materials.

8. The use according to any one of claims 1 to 7, characterized in that: The composition is prepared into a traditional Chinese medicine oral preparation.

9. The use according to any one of claims 8, characterized in that The Chinese medicine oral preparations are granules, oral solutions, mixtures, pills, syrups, capsules and tablets.

10. The use according to any one of claims 9, characterized in that: The Chinese medicine oral preparation is a mixture or granules.

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