Use of a xanthanamide in the preparation of a medicament for the treatment of multiple sclerosis

Abstract

The application discloses application of Zanthoxylum amide in preparation of a drug for treating multiple sclerosis. The Zanthoxylum amide is selected from hydroxyl-alpha- toosendanin and hydroxyl-beta-toosendanin. Research results of the application show that hydroxyl-alpha-toosendanin and / or hydroxyl-beta-toosendanin has a good treatment effect on an experimental autoimmune encephalomyelitis mouse model. Compared with a model group, drug intervention can obviously improve nerve dysfunction and disability degree of the mouse, delay clinical symptoms, reduce inflammation of spinal cord tissue, regulate immune function, and show a good treatment effect. The application can provide a new drug for treating multiple sclerosis.

Description

Application of a type of cinnamamide in the preparation of a drug for treating multiple sclerosis Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, specifically, it relates to the application of zanthoxylate in the preparation of a drug for treating multiple sclerosis. Background Technology

[0002] Multiple sclerosis (MS) is an autoimmune disease characterized by inflammatory demyelinating lesions of the white matter in the central nervous system. Its main manifestations include decreased vision, diplopia, sensory disturbances in the limbs, motor disturbances in the limbs, and bladder dysfunction. The disease has a temporal and spatial multifocal pattern, commonly affecting young and middle-aged adults, and has become the most common cause of permanent disability in young adults besides trauma. Currently, there is no unified and effective treatment for MS. Clinically, corticosteroids such as prednisone are commonly used to relieve acute symptoms. Drugs such as beta-interferon, glamer acetate, natezumab, and fingolimod are used to prevent the immune system from attacking the myelin sheath to reduce the number of future relapses. Other drugs that relieve or control specific symptoms are also used to improve survival. While corticosteroids can shorten the duration of attacks, they have significant side effects and are not suitable for long-term use. Immunomodulatory agents such as beta-interferon and natezumab are expensive, and long-term use of single-target drugs can easily lead to tolerance and increase the risk of progressive multifocal leukoencephalopathy. Traditional Chinese medicine (TCM) has unique advantages in the treatment of multiple sclerosis (MS) due to its fewer adverse reactions, multiple targets, multiple pathways, and systemic regulatory effects. Therefore, it is of great significance to discover safe and effective anti-MS drugs with fewer side effects from the treasure trove of traditional Chinese medicine. Summary of the Invention

[0003] The purpose of this invention is to provide the application of zanthamide in the preparation of a medicament for treating multiple sclerosis.

[0004] To achieve the above objectives, the technical solution adopted by the present invention is as follows:

[0005] The first aspect of the present invention provides the use of zanthamide in the preparation of a medicament for treating multiple sclerosis.

[0006] The zanthoxyl amide is selected from hydroxy-α-zanthoxyl and hydroxy-β-zanthoxyl (Reference: Jingjing Luo, Jingxuan Ke, Xiaoyan Hou, Shanshan Li, Qingying Luo, Hejun Wu, Guanghui Shen, Zhiqing Zhang, Composition, structure and flavor mechanism of numbing substances in Chinese prickly ash in the genus Zanthoxylum: A review, Food Chemistry, 2022, 373(Pt B):131454.).

[0007] The structure of the hydroxy-α-sanshool is as follows:

[0008]

[0009] The structure of the hydroxy-β-sanshool is as follows:

[0010]

[0011] The drug mentioned for treating multiple sclerosis refers to a drug with citric acid amide as its sole active ingredient.

[0012] The dosage form of the drug for treating multiple sclerosis is capsule, tablet, granule, solution, suspension, emulsion or syrup.

[0013] The medication for treating multiple sclerosis is administered orally.

[0014] By adopting the above technical solution, the present invention has the following advantages and beneficial effects:

[0015] The results of this invention show that hydroxy-α-sanshool and / or hydroxy-β-sanshool have a good therapeutic effect on an experimental autoimmune encephalomyelitis (EAE) mouse model. Compared with the model group, drug intervention can significantly improve the degree of neurological dysfunction and disability in mice, delay clinical symptoms, and reduce spinal cord inflammation, demonstrating a good therapeutic effect. This invention can provide a new drug for the treatment of multiple sclerosis.

[0016] This invention presents a systematic chemical study on the amide components of Zanthoxylum plants, isolating a large number of amide compounds and conducting extensive activity studies. Experiments revealed that hydroxy-α-sanshool and / or hydroxy-β-sanshool possess good anti-multiple sclerosis effects, suggesting potential new applications and significant clinical value.

[0017] Animal efficacy experiments of this invention have confirmed that the zanthoxylate compounds hydroxy-α-sanshool and / or hydroxy-β-sanshool can effectively improve nerve function, alleviate symptoms such as paralysis and disability, and inhibit spinal cord inflammation, thereby achieving the goal of treating multiple sclerosis. The above-mentioned substances hydroxy-α-sanshool and / or hydroxy-β-sanshool are widely available and can provide new drugs for the treatment of multiple sclerosis, possessing significant clinical application value. Attached Figure Description

[0018] Figure 1 is a schematic diagram showing the effects of hydroxy-α-sanshool and hydroxy-β-sanshool treatment on the clinical symptoms of the EAE multiple sclerosis mouse model.

[0019] Figure 2 is a schematic diagram showing the effects of hydroxy-α-sanshool and hydroxy-β-sanshool treatment on spinal cord tissue inflammation in an EAE multiple sclerosis mouse model.

[0020] Figure 3 is a schematic diagram showing the effect of hydroxy-α-sanshool tablets and hydroxy-β-sanshool tablets on the clinical symptoms of EAE multiple sclerosis mouse model.

[0021] Figure 4 is a schematic diagram showing the effect of hydroxy-α-sanshool tablets and hydroxy-β-sanshool tablets on spinal cord tissue inflammation in EAE multiple sclerosis mouse models. Detailed Implementation

[0022] To more clearly illustrate the present invention, the following description, in conjunction with preferred embodiments, further clarifies the invention. Those skilled in the art should understand that the specific descriptions below are illustrative rather than restrictive, and should not be construed as limiting the scope of protection of the present invention.

[0023] Example 1

[0024] Establishment and drug intervention of a mouse model of multiple sclerosis

[0025] Experimental autoimmune encephalomyelitis (EAE) mouse models are a commonly used classic model in multiple sclerosis research, closely resembling the key pathological features of immune inflammation and demyelination associated with the disease. This invention uses MOG / Freud's complete adjuvant / pertussis toxin to induce this model.

[0026] Establishment and grouping of the EAE mouse model: After one week of acclimatization, male C57BL / 6 mice were randomly divided into four groups: Normal, Model, Low-dose hydroxy-α-sanshool (HASL, 10 mg / kg), High-dose hydroxy-α-sanshool (HASH, 20 mg / kg), Low-dose hydroxy-β-sanshool (HBSL, 10 mg / kg), and High-dose hydroxy-β-sanshool (HBSH, 20 mg / kg), with 10 mice in each group. Except for the Normal group, the other groups received subcutaneous injections of 50 μL of antigen emulsion (containing MOG) at four points on both sides of the spine on the back of the mice. 35-55 250 μg of antigen, 800 μg of inactivated Mycobacterium tuberculosis H37RA, 100 μL of PBS, and 100 μL of Freund's complete adjuvant were administered to mice via intraperitoneal injection. 100 μL (500 ng / mouse) of pertussis toxin was injected intraperitoneally 30 min after antigen emulsion injection and again on the second day. Each treatment group was administered the corresponding drug via gavage starting from the first day of modeling, at a volume of 10 mL / kg, once daily. The normal and model groups were administered the same volume of distilled water via gavage for four consecutive weeks. During the experiment, the mice's disease progression was observed at a fixed time each morning, and neurological function was assessed using the KONO'S scoring method. For the last three days, coordination and balance were assessed using a variable-speed rotarod fatigue tester. After the experiment, the mice were sacrificed, and spinal cord tissue was extracted from each group. The levels of interferon-γ (IFN-γ), tumor necrosis factor-α (TNF-α), and interleukin-17A (IL-17A) were detected using an ELISA kit according to the manufacturer's instructions.

[0027] The results are analyzed as follows:

[0028] Figure 1 illustrates the effects of hydroxy-α-sanshool and hydroxy-β-sanshool treatment on clinical symptoms in a mouse model of EAE (Extra-Age Emission) and multiple sclerosis. In the figure, A represents the KONO'S clinical score. During the experiment, the normal group mice showed good growth, active eating, and glossy fur without abnormalities. The model group mice began to exhibit irritability on day 4 and began to develop symptoms on day 8, including limb weakness, lameness, and paralysis. The mice in each treatment group began to show disease-related symptoms such as tail weakness and lameness on day 12. Compared with the model group, intervention with hydroxy-α-sanshool and hydroxy-β-sanshool (10 and 20 mg / kg) significantly reduced EAE induction, delayed clinical symptoms, and lowered clinical scores.

[0029] The coordination and balance of mice were tested using a rotating bar, and the duration of the rod stay was recorded, as shown in Figure 1B. B is a schematic diagram of the rod time results. The results showed that the average rod time of the model group mice was significantly reduced (p<0.01). Oral administration of hydroxy-α-sanshool and hydroxy-β-sanshool (10, 20 mg / kg) could significantly improve the limb motor balance dysfunction caused by EAE (p<0.05 or p<0.01).

[0030] IFN-γ, IL-17A, and TNF-α are common pro-inflammatory cytokines in the body. The results are shown in Figure 2. Figure 2 is a schematic diagram illustrating the effects of hydroxy-α-salicornin and hydroxy-β-salicornin treatment on spinal cord inflammation in an EAE multiple sclerosis (MS) mouse model. Specifically, A shows the effect of hydroxy-α-salicornin and hydroxy-β-salicornin treatment on IFN-γ inflammation in the spinal cord of the EAE MS mouse model; B shows the effect of hydroxy-α-salicornin and hydroxy-β-salicornin treatment on IL-17A inflammation in the spinal cord of the EAE MS mouse model; and C shows the effect of hydroxy-α-salicornin and hydroxy-β-salicornin treatment on TNF-α inflammation in the spinal cord of the EAE MS mouse model. IFN-γ and IL-17A can increase blood-brain barrier permeability and are related to the pathogenesis of MS; TNF-α and its soluble receptors are associated with disability, disability progression, and clinical manifestations of MS. The levels of the above three substances in spinal cord tissue were measured by ELISA. The results showed that, compared with the normal group of mice, the levels of IFN-γ, IL-17A and TNF-α in the spinal cord tissue of the model group of mice were significantly increased (p<0.01). The levels of these inflammatory factors in the tissues of mice that were administered low- and high-dose drugs by gavage were significantly reduced (p<0.05 or p<0.01), suggesting that hydroxy-α-sanshool and hydroxy-β-sanshool have a good effect in inhibiting spinal cord immune inflammation.

[0031] In summary, hydroxy-α-salicornin and / or hydroxy-β-salicornin can inhibit spinal cord inflammation, significantly improve the neurological function and overall functional status of EAE model mice, and inhibit the occurrence and development of multiple sclerosis, showing great potential as drugs for the treatment of multiple sclerosis.

[0032] Example 2

[0033] Tablet preparation:

[0034] 5g of hydroxy-α-sanshool or hydroxy-β-sanshool

[0035] 42g of lactose

[0036] 12g corn starch

[0037] 1g magnesium stearate

[0038] Preparation method: Mix hydroxy-α-sanshool or hydroxy-β-sanshool, lactose, and corn starch. Moisten the mixture evenly with 20 mL of water, sieve the mixture, dry it at 55°C, sieve it again, add magnesium stearate, mix well, sieve again, and compress it into tablets using a single-punch tablet press. Each tablet weighs 300 mg and contains 25 mg of hydroxy-α-sanshool or hydroxy-β-sanshool.

[0039] Pharmacodynamic studies of tablets:

[0040] Establishment and grouping of the EAE mouse model: After one week of acclimatization, male C57BL / 6 mice were randomly divided into three groups: Normal, Model, hydroxy-α-sanshool tablet group (HAST, 48 mg / kg), and hydroxy-β-sanshool tablet group (HBST, 48 mg / kg), with 10 mice in each group. Except for the Normal group, the other groups received subcutaneous injections of 50 μL of antigen emulsion (containing MOG) at four points on both sides of the spine on the back of the mice. 35-55 250 μg of antigen, 800 μg of inactivated Mycobacterium tuberculosis H37RA, 100 μL of PBS, and 100 μL of Freund's complete adjuvant were administered to mice via intraperitoneal injection. 100 μL (500 ng / mouse) of pertussis toxin was injected intraperitoneally 30 min after antigen emulsion injection and again on the second day. Each treatment group was administered the corresponding drug via gavage starting from the first day of modeling, at a volume of 10 mL / kg, once daily. The normal and model groups were administered the same volume of distilled water via gavage for four consecutive weeks. During the experiment, the mice's disease progression was observed at a fixed time each morning, and neurological function was assessed using the KONO'S scoring method. For the last three days, coordination and balance were assessed using a variable-speed rotarod fatigue tester. After the experiment, the mice were sacrificed, and spinal cord tissue from each group was extracted. The levels of IFN-γ, TNF-α, and IL-17A were detected using an ELISA kit according to the manufacturer's instructions.

[0041] The results are shown in Figure 3, which illustrates the effects of hydroxy-α-sanshool tablets and hydroxy-β-sanshool tablets on the clinical symptoms of EAE multiple sclerosis mouse models. In the figure, A represents the KONO'S clinical score. During the experiment, the normal group mice showed good growth, glossy fur, and normal activity. The model group mice began to exhibit irritability on day 5 and began to develop symptoms on day 8, including tail weakness, hind limb dragging, and limb paralysis. The mice in each treatment group began to show disease-related symptoms such as tail weakness and lameness on day 12. Compared with the model group, intervention with hydroxy-α-sanshool tablets and hydroxy-β-sanshool tablets significantly reduced EAE induction, delayed clinical symptoms, and lowered clinical scores. The coordination and balance of mice were tested using a rotating bar, and the duration of the mice's stay was recorded, as shown in Figure 3B. The average time of the model group mice on the bar was significantly reduced (p<0.01). After gavage administration of hydroxy-α-sanshool tablets and hydroxy-β-sanshool tablets, the limb motor balance dysfunction caused by EAE was significantly improved (p<0.01).

[0042] The results are shown in Figure 4. Figure 4 is a schematic diagram of the effects of hydroxy-α-sanshozoline tablets and hydroxy-β-sanshozoline tablets on spinal cord inflammation in EAE multiple sclerosis mouse models. Specifically, A shows the effect of gavage administration of hydroxy-α-sanshozoline tablets and hydroxy-β-sanshozoline tablets on IFN-γ inflammation in the spinal cord of the EAE multiple sclerosis mouse model; B shows the effect of gavage administration of hydroxy-α-sanshozoline tablets and hydroxy-β-sanshozoline tablets on IL-17A inflammation in the spinal cord of the EAE multiple sclerosis mouse model; and C shows the effect of gavage administration of hydroxy-α-sanshozoline tablets and hydroxy-β-sanshozoline tablets on TNF-α inflammation in the spinal cord of the EAE multiple sclerosis mouse model. The results showed that, compared with the normal group mice, the levels of IFN-γ, IL-17A and TNF-α in the spinal cord tissue of the model group mice were significantly increased (p<0.01). Mice administered hydroxy-α-sanshool tablets or hydroxy-β-sanshool tablets by gavage showed significantly decreased levels of these inflammatory factors in their tissues (p<0.05 or p<0.01), suggesting that hydroxy-α-sanshool tablets and hydroxy-β-sanshool tablets have a good effect in inhibiting spinal cord immune inflammation.

[0043] The above results indicate that the hydroxy-α-sanshool tablets and hydroxy-β-sanshool tablets can reduce the severity of multiple sclerosis, improve spinal cord inflammation, and have a good therapeutic effect on the disease.

[0044] Example 3

[0045] Preparation of oral suspensions:

[0046] 6g of hydroxy-α-sanshool or hydroxy-β-sanshool

[0047] Citric acid 1g

[0048] 20 mL of glycerol

[0049] Tween-80 2mL

[0050] 0.2g of pregelatinized starch

[0051] Sodium benzoate 0.2g

[0052] Xanthan gum 0.5g

[0053] Add purified water to 200 mL.

[0054] Preparation method: Take hydroxy-α-sanshool or hydroxy-β-sanshool, add Tween-80 and glycerol, mix thoroughly, and then add pregelatinized starch, xanthan gum, sodium benzoate and citric acid while stirring. After it is completely uniform, add water to 200mL and stir to disperse it fully to obtain an oral suspension.

[0055] The above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention in any way. Although the present invention has been disclosed above with reference to preferred embodiments, it is not intended to limit the present invention. Any person skilled in the art can make some modifications or alterations to the above-described technical content to create equivalent embodiments without departing from the scope of the present invention. Any simple modifications, equivalent changes, and alterations made to the above embodiments based on the technical essence of the present invention without departing from the scope of the present invention shall still fall within the scope of the present invention.

Claims

1. The use of a type of xanthanamide in the preparation of a medicament for treating multiple sclerosis, characterized in that, The zanthamide is selected from hydroxy-α-santhiocin and hydroxy-β-santhiocin; the structure of hydroxy-α-santhiocin is as follows: The structure of the hydroxy-β-sanshool is as follows: The drug mentioned for treating multiple sclerosis refers to a drug with citric acid amide as its sole active ingredient.

2. The use of the zanthoxylans according to claim 1 in the preparation of a medicament for treating multiple sclerosis, characterized in that, The dosage form of the drug for treating multiple sclerosis is capsule, tablet, granule, solution, suspension, emulsion or syrup.

3. The use of the zanthamide according to claim 1 in the preparation of a medicament for treating multiple sclerosis, characterized in that, The medication for treating multiple sclerosis is administered orally.

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