Drug combination and use thereof in prevention or treatment of neurodegenerative disease

Abstract

A drug combination comprising withaferin A and use thereof. The use comprises separate administration or combined administration of withaferin A or a salt thereof and pitolisant or a salt thereof. The combination therapy with withaferin A or the salt thereof and pitolisant or the salt thereof can prevent and treat a neurodegenerative disease, effectively protect neurons, and treat neuroinflammation. The combined administration can not only significantly improve object recognition ability, spatial memory, and working memory, but can also achieve effects superior to those of separate administration.

Description

A drug combination and its use in preventing or treating neurodegenerative diseases Technical Field

[0001] The present disclosure belongs to the field of medicine and biological technology, and relates to the use of a drug combination in preventing or treating neurodegenerative diseases, and in particular to a combination drug containing withaferin A and tilorixan and its use in preventing or treating neurodegenerative diseases. Background Art

[0002] Neurodegenerative diseases are a group of neurological disorders characterized by progressive damage to neurons in the central and peripheral nervous systems, including Alzheimer's disease, Parkinson's disease, and Huntington's disease. Damage to the structure and function of neural networks and neuronal loss lead to the breakdown of neural circuits, ultimately manifesting as impairments in memory, cognition, behavior, sensation, and motor function. The causes of these diseases remain unclear, and there is no effective cure. These diseases pose a serious threat to the quality of life for millions of patients worldwide.

[0003] Alzheimer's disease (AD) is an age-related neurodegenerative disorder with an unclear etiology and no effective cure. With the aging of the global population, the incidence of AD is increasing significantly. Furthermore, AD poses a significant threat to the physical and mental health of patients and places a heavy economic burden on families and society, making it a serious public health issue.

[0004] Currently, the primary goal of treating neurodegenerative diseases is to slow disease progression and control symptoms. Treatment options include medications to alleviate the progression and severity of the disease. While medications can alleviate symptoms in most patients, they cannot halt disease progression. These medications often lead to discontinuation due to significant side effects, or their effectiveness diminishes with prolonged use. Therefore, finding an effective drug to prevent and treat neurodegenerative diseases and improve patients' quality of life is an urgent need. Summary of the Invention

[0005] One aspect of the present disclosure provides a pharmaceutical composition comprising i) withaferin A or a salt thereof; and ii) tilorixan or a salt thereof.

[0006] In some embodiments, the pharmaceutical composition of the present disclosure comprises a therapeutically effective amount of withaferin A or a salt thereof, a therapeutically effective amount of tilorixan or a salt thereof, and a pharmaceutically acceptable carrier or excipient.

[0007] In some embodiments, the pharmaceutical composition of the present disclosure comprises about 1 mg to 4000 mg of withaferin A or a salt thereof, preferably about 50 mg to 200 mg of withaferin A or a salt thereof, preferably about 50 mg to 100 mg of withaferin A or a salt thereof, preferably about 100 mg to 200 mg of withaferin A or a salt thereof, preferably about 200 mg to 400 mg of withaferin A or a salt thereof. The pharmaceutical composition can be formulated in a unit dosage form.

[0008] In some embodiments, the pharmaceutical composition of the present disclosure comprises about 1 mg to 4000 mg of tilorexant or a salt thereof, preferably about 50 mg to 200 mg of tilorexant or a salt thereof, preferably about 50 mg to 100 mg of tilorexant or a salt thereof, preferably about 100 mg to 200 mg of tilorexant or a salt thereof, preferably about 200 mg to 400 mg of tilorexant or a salt thereof, preferably about 400 mg to 1000 mg of tilorexant or a salt thereof. The pharmaceutical composition can be formulated in a unit dosage form.

[0009] In some embodiments, the dosage ratio of withaferin A or its salt and tilorixan or its salt in the pharmaceutical composition of the present disclosure is about 10:1 to about 1:10, preferably about 2:1 to about 1:2, and more preferably about 1:1 or 1:2.

[0010] In some embodiments, the pharmaceutical compositions of the present disclosure are used to prevent or treat neurodegenerative diseases.

[0011] In some embodiments, the neurodegenerative disease is selected from Alzheimer's disease, Parkinson's disease, or Huntington's disease. That is, in some embodiments, the pharmaceutical composition of the present disclosure is used to prevent or treat Alzheimer's disease, Parkinson's disease, or Huntington's disease.

[0012] Another aspect of the present disclosure provides use of a pharmaceutical composition in preparing a medicament for preventing or treating a neurodegenerative disease in a subject, the pharmaceutical composition comprising i) withaferin A or a salt thereof; and ii) tilorixen or a salt thereof.

[0013] In some embodiments, the pharmaceutical composition according to the present disclosure is used in the preparation of a medicament for preventing or treating a neurodegenerative disease in a subject, wherein the pharmaceutical composition comprises a therapeutically effective amount of withaferin A or a salt thereof, a therapeutically effective amount of tilorixan or a salt thereof, and a pharmaceutically acceptable carrier or excipient.

[0014] In some embodiments, the pharmaceutical composition according to the present disclosure is used in the preparation of a medicament for preventing or treating a neurodegenerative disease in a subject, wherein the pharmaceutical composition comprises about 1 mg to 4000 mg of withaferin A or a salt thereof, preferably about 50 mg to 200 mg of withaferin A or a salt thereof, preferably about 50 mg to 100 mg of withaferin A or a salt thereof, preferably about 100 mg to 200 mg of withaferin A or a salt thereof, preferably about 200 mg to 400 mg of withaferin A or a salt thereof. The pharmaceutical composition can be formulated in a unit dosage form.

[0015] In some embodiments, the pharmaceutical composition according to the present disclosure is used in the preparation of a medicament for preventing or treating a neurodegenerative disease in a subject, wherein the pharmaceutical composition comprises about 1 mg to 4000 mg of tilorexan or a salt thereof, preferably about 50 mg to 200 mg of tilorexan or a salt thereof, preferably about 50 mg to 100 mg of tilorexan or a salt thereof, preferably about 100 mg to 200 mg of tilorexan or a salt thereof, preferably about 200 mg to 400 mg of tilorexan or a salt thereof, preferably about 400 mg to 1000 mg of tilorexan or a salt thereof. The pharmaceutical composition can be formulated in a unit dosage form.

[0016] In some embodiments, the pharmaceutical composition according to the present disclosure is used in the preparation of a medicament for preventing or treating a neurodegenerative disease in a subject, wherein the dosage ratio of i) withaferin A or a salt thereof and ii) tilorixen or a salt thereof in the pharmaceutical composition is about 10:1 to about 1:10, preferably about 2:1 to about 1:2, and more preferably about 1:1 or 1:2.

[0017] In some embodiments, the neurodegenerative disease is selected from Alzheimer's disease, Parkinson's disease, or Huntington's disease. That is, in some embodiments, the present disclosure provides a use of a pharmaceutical composition in the preparation of a medicament for preventing or treating Alzheimer's disease, Parkinson's disease, or Huntington's disease.

[0018] Another aspect of the present disclosure provides use of withaferin A or a salt thereof in preparing a combination pharmaceutical composition for preventing or treating a neurodegenerative disease in a subject, the combination pharmaceutical composition comprising i) withaferin A or a salt thereof; and ii) tilorixan or a salt thereof.

[0019] Another aspect of the present disclosure provides use of tilorixan or a salt thereof in preparing a combination pharmaceutical composition for preventing or treating a neurodegenerative disease in a subject, the combination pharmaceutical composition comprising i) withaferin A or a salt thereof; and ii) tilorixan or a salt thereof.

[0020] In some embodiments, the use of withaferin A or a salt thereof and tilorixan or a salt thereof according to the present disclosure in the preparation of a combination pharmaceutical composition for preventing or treating a neurodegenerative disease in a subject, wherein the combination pharmaceutical composition comprises a therapeutically effective amount of withaferin A or a salt thereof, a therapeutically effective amount of tilorixan or a salt thereof, and a pharmaceutically acceptable carrier or excipient.

[0021] In some embodiments, the use of withaferin A or a salt thereof and tilorixan or a salt thereof according to the present disclosure in the preparation of a combination pharmaceutical composition for preventing or treating a neurodegenerative disease in a subject, wherein the pharmaceutical composition comprises about 1 mg to 4000 mg of withaferin A or a salt thereof. The pharmaceutical composition preferably comprises about 50 mg to 200 mg of withaferin A or a salt thereof, preferably about 50 mg to 100 mg of withaferin A or a salt thereof, preferably about 100 mg to 200 mg of withaferin A or a salt thereof, preferably about 200 mg to 400 mg of withaferin A or a salt thereof. The pharmaceutical composition can be formulated in a unit dosage form.

[0022] In some embodiments, according to the present disclosure, withaferin A or a salt thereof and tilorexin or a salt thereof are used in the preparation of a combination pharmaceutical composition for preventing or treating a neurodegenerative disease in a subject, wherein the pharmaceutical composition comprises about 1 mg to 4000 mg of tilorexin or a salt thereof. The pharmaceutical composition comprises about 1 mg to 4000 mg of tilorexin or a salt thereof, preferably about 50 mg to 200 mg of tilorexin or a salt thereof, preferably about 50 mg to 100 mg of tilorexin or a salt thereof, preferably about 100 mg to 200 mg of tilorexin or a salt thereof, preferably about 200 mg to 400 mg of tilorexin or a salt thereof, preferably about 400 mg to 1000 mg of tilorexin or a salt thereof. The pharmaceutical composition can be formulated in a unit dosage form.

[0023] In some embodiments, the use of withaferin A or a salt thereof and tilorixan or a salt thereof according to the present disclosure in preparing a combination pharmaceutical composition for preventing or treating a neurodegenerative disease in a subject, wherein in the pharmaceutical composition, the mass ratio of i) withaferin A or a salt thereof and ii) tilorixan or a salt thereof is about 10:1 to about 1:10, preferably about 2:1 to about 1:2, and more preferably about 1:1 or 1:2.

[0024] In some embodiments, the neurodegenerative disease is selected from Alzheimer's disease, Parkinson's disease, or Huntington's disease. That is, in some embodiments, the present disclosure provides the use of withaferin A or a salt thereof and tilorixan or a salt thereof in the preparation of a combination pharmaceutical composition for preventing or treating Alzheimer's disease, Parkinson's disease, or Huntington's disease.

[0025] Another aspect of the present disclosure provides a method for preventing or treating a neurodegenerative disease in a subject in need thereof, comprising administering to a subject in need thereof a therapeutically effective amount of withaferin A or a salt thereof, and administering to the subject a therapeutically effective amount of tilorixan or a salt thereof.

[0026] In some embodiments, the present disclosure provides a method for preventing or treating a neurodegenerative disease in a subject in need thereof, wherein the administration of withaferin A or a salt thereof and tilorixen or a salt thereof is performed simultaneously or sequentially in any order.

[0027] In some embodiments, the method of preventing or treating a neurodegenerative disease in a subject in need thereof is disclosed herein, wherein withaferin A or a salt thereof is administered in an amount of about 0.02-40 mg / kg, preferably in an amount of about 2-4 mg / kg, or about 1 mg / kg to about 2 mg / kg, daily or once every two days.

[0028] In some embodiments, the present disclosure provides a method for preventing or treating a neurodegenerative disease in a subject in need thereof, wherein tilorixen or a salt thereof is administered in an amount of about 0.02-40 mg / kg, preferably about 2-10 mg / kg, preferably about 1 mg / kg to about 2 mg / kg, daily or once every two days.

[0029] In some embodiments, the present disclosure provides a method for preventing or treating a neurodegenerative disease in a subject in need thereof, wherein the administered dosage ratio of i) withaferin A or a salt thereof and ii) tilorixan or a salt thereof is about 10:1 to about 1:10, preferably about 1:1 or 1:2.

[0030] The present disclosure also provides the use of withaferin A or a salt thereof in enhancing the effect of tilorexin or a salt thereof in preventing or treating neurodegenerative diseases in a subject. Specifically, the use of withaferin A or a salt thereof in preparing a potentiator for tilorexin or a salt thereof in preventing or treating neurodegenerative diseases.

[0031] According to an embodiment of the present disclosure, the amount of withaferin A or its salt is as defined herein.

[0032] The present disclosure also provides the use of tilorixan or a salt thereof in enhancing the effect of withaferin A or a salt thereof in preventing or treating neurodegenerative diseases in a subject. Specifically, the use of tilorixan or a salt thereof in preparing a potentiator for withaferin A or a salt thereof in preventing or treating neurodegenerative diseases.

[0033] According to an embodiment of the present disclosure, the amount of tilorixen or its salt is as defined herein.

[0034] According to an embodiment of the present disclosure, the dosage ratio of withaferin A or its salt and tilorixan or its salt is as defined herein.

[0035] According to an embodiment of the present disclosure, the neurodegenerative disease is selected from Alzheimer's disease, Parkinson's disease, or Huntington's disease.

[0036] In some embodiments, the disclosed methods of preventing or treating a neurodegenerative disease in a subject in need thereof are selected from Alzheimer's disease, Parkinson's disease, or Huntington's disease.

[0037] In some embodiments, the subject of the present disclosure is a mammal, preferably a human subject.

[0038] The above aspects and embodiments and other aspects, objects, features and advantages of the present disclosure will become apparent from the following detailed description. BRIEF DESCRIPTION OF THE DRAWINGS

[0039] Figure 1 shows the results of the novel object recognition experiment in mice in the Alzheimer's disease model group and the control group, which were intraperitoneally injected with control solvent, 2 mg / kg withaferin A, 2 mg / kg tilorexant, 1 mg / kg withaferin A + 1 mg / kg tilorexant, and 2 mg / kg withaferin A + 2 mg / kg tilorexant, respectively.

[0040] Figure 2 shows the results of the novel location recognition experiment in mice in the Alzheimer's disease model group and the control group, which were intraperitoneally injected with control solvent, 2 mg / kg withaferin A, 2 mg / kg tilorexant, 1 mg / kg withaferin A + 1 mg / kg tilorexant, and 2 mg / kg withaferin A + 2 mg / kg tilorexant, respectively.

[0041] Figure 3 shows the correct choice rate (%) of the T maze test in mice at week 4 after intraperitoneal injection of control solvent, 2 mg / kg withaferin A, 2 mg / kg tilorexant, 1 mg / kg withaferin A + 1 mg / kg tilorexant, and 2 mg / kg withaferin A + 2 mg / kg tilorexant in the Alzheimer's disease model group and the control group, respectively.

[0042] Figure 4 shows the choice latency time of the T-maze training phase of mice in the Alzheimer's disease model group and the control group, which were intraperitoneally injected with control solvent, 2 mg / kg withaferin A, 2 mg / kg tilorexant, 1 mg / kg withaferin A + 1 mg / kg tilorexant, and 2 mg / kg withaferin A + 2 mg / kg tilorexant, respectively, at week 4.

[0043] Figure 5 shows the body weight changes in the T-maze test of mice in the Alzheimer's disease model group and the control group after intraperitoneal injection of control solvent, 2 mg / kg withaferin A, 2 mg / kg tilorexant, 1 mg / kg withaferin A + 1 mg / kg tilorexant, and 2 mg / kg withaferin A + 2 mg / kg tilorexant, respectively.

[0044] Figure 6 shows the correct choice rate (%) and choice delay time of the T-maze test phase of mice in the Alzheimer's disease model group and the control group, which were intraperitoneally injected with control solvent, 2 mg / kg withaferin A, 2 mg / kg tilorexant, 1 mg / kg withaferin A + 1 mg / kg tilorexant, and 2 mg / kg withaferin A + 2 mg / kg tilorexant, respectively, at week 4. DETAILED DESCRIPTION

[0045] In one aspect of the present disclosure, provided are effects of withaferin A for improving cognitive function, and its effects for preventing or treating neurodegenerative diseases, preferably for preventing or treating Alzheimer's disease and / or Parkinson's disease, more preferably for preventing or treating Alzheimer's disease.

[0046] Another aspect of the present disclosure provides the effects of tilorixen for improving cognitive function, and its effects for preventing or treating neurodegenerative diseases, preferably for preventing or treating Alzheimer's disease, Parkinson's disease and / or Huntington's disease, more preferably for preventing or treating Alzheimer's disease.

[0047] Another aspect of the present disclosure provides a pharmaceutical composition for use in improving cognitive function and preventing or treating neurodegenerative diseases, preferably Alzheimer's disease, Parkinson's disease, or Huntington's disease, and more preferably Alzheimer's disease. The pharmaceutical composition comprises withaferin A or a salt thereof and tilorixan or a salt thereof as active ingredients, and further comprises pharmaceutically acceptable excipients or auxiliary ingredients.

[0048] Another aspect of the present disclosure provides a combination drug and its use for improving cognitive function, particularly in the prevention or treatment of neurodegenerative diseases, preferably for the prevention or treatment of Alzheimer's disease, Parkinson's disease, or Huntington's disease, and more preferably for the prevention or treatment of Alzheimer's disease. The combination drug comprises withaferin A or its salt and tilorixan or its salt as active ingredients and further comprises pharmaceutically acceptable excipients or auxiliary ingredients. In a specific embodiment, the withaferin A or its salt and tilorixan or its salt can be administered to the subject together, or separately and sequentially in any order.

[0049] According to an embodiment of the present disclosure, the use of withaferin A or a salt thereof and / or tilorixan or a salt thereof in the preparation of a medicine, a food or a health product is proposed, and the medicine, the food or the health product is used to improve cognitive function.

[0050] According to an embodiment of the present disclosure, the use of withaferin A or a salt thereof and / or tilorixan or a salt thereof in the preparation of medicines, foods or health products is proposed. The medicines, foods or health products are used to prevent or treat neurodegenerative diseases, preferably to prevent or treat Alzheimer's disease, Parkinson's disease, or Huntington's disease, and more preferably to prevent or treat Alzheimer's disease.

[0051] The dosage form of the drug preferably includes one or more of tablets, powders, granules, capsules, oral solutions and sustained-release preparations.

[0052] According to the present disclosure, the amount of withaferin A or its salt can be about 0.02 mg / kg to about 40 mg / kg, preferably about 2 mg / kg to about 4 mg / kg, more preferably about 1 mg / kg to about 2 mg / kg; the amount of tilorixan or its salt can be about 0.02 mg / kg to about 40 mg / kg, preferably about 2 mg / kg to about 40 mg / kg, preferably about 2 mg / kg to about 10 mg / kg, preferably about 2 mg / kg to about 4 mg / kg or about 4 mg / kg to about 8 mg / kg. It can be administered daily or once every two days.

[0053] According to the present disclosure, the dosage ratio of withaferin A or its salt and tilorixan or its salt is about 10:1 to about 1:10, preferably about 1:1 or 1:2.

[0054] According to an embodiment of the present disclosure, preferably, the pharmaceutical composition or combination drug of 2 mg / kg withaferin A or its salt and 2 mg / kg tilorixan or its salt can prevent or treat neurodegenerative diseases, and can achieve better effects than using either alone.

[0055] The present disclosure also provides a pharmaceutical composition or a combination drug containing withaferin A or a salt thereof and tilorixan or a salt thereof.

[0056] According to an embodiment of the present disclosure, the pharmaceutical composition or combination drug of the present disclosure comprises about 1 mg to 4000 mg of withaferin A or a salt thereof. The pharmaceutical composition preferably comprises about 50 mg to 200 mg of withaferin A or a salt thereof, preferably about 50 mg to 100 mg of withaferin A or a salt thereof, preferably about 100 mg to 200 mg of withaferin A or a salt thereof, preferably about 200 mg to 400 mg of withaferin A or a salt thereof. The pharmaceutical composition can be formulated in a unit dosage form.

[0057] According to an embodiment of the present disclosure, in the pharmaceutical composition or combination drug of the present disclosure, the pharmaceutical composition comprises about 1 mg to 4000 mg of tilorexant or a salt thereof, preferably about 50 mg to 200 mg of tilorexant or a salt thereof, preferably about 50 mg to 100 mg of tilorexant or a salt thereof, preferably about 100 mg to 200 mg of tilorexant or a salt thereof, preferably about 200 mg to 400 mg of tilorexant or a salt thereof, preferably about 400 mg to 1000 mg of tilorexant or a salt thereof. The pharmaceutical composition can be formulated in a unit dosage form.

[0058] According to an embodiment of the present disclosure, the pharmaceutical composition or combination drug of the present disclosure comprises a therapeutically effective amount of withaferin A or a salt thereof, a therapeutically effective amount of tilorixan or a salt thereof, and a pharmaceutically acceptable carrier or excipient.

[0059] According to an embodiment of the present disclosure, in the pharmaceutical composition or combination drug of the present disclosure, the dosage ratio of withaferin A or its salt and tilorixan or its salt is about 10:1 to 1:10, preferably about 2:1 to about 1:2, and more preferably about 1:1 or 1:2.

[0060] According to embodiments of the present disclosure, pharmaceutical compositions or combination drugs containing withaferin A or its salt and tilorixan or its salt can be prepared in a unit dosage form to facilitate drug production and administration. Therefore, the present disclosure also provides a unit dosage form comprising withaferin A or its salt and tilorixan or its salt in the amounts described above.

[0061] The present disclosure also provides a pharmaceutical composition or a combination drug containing withaferin A or a salt thereof and tilorixan or a salt thereof, for preventing or treating neurodegenerative diseases.

[0062] According to an embodiment of the present disclosure, object recognition memory is detected by novel object recognition (NORT); spatial memory is detected by novel location recognition (NLRT); and spatial working memory is detected by T-maze.

[0063] The present disclosure also provides the use of the pharmaceutical composition or combination drug described in the above technical solution in the preparation of a drug for preventing or treating neurodegenerative diseases in a subject. In the present disclosure, the pharmaceutical composition or combination drug preferably contains a therapeutically effective amount of withaferin A or its salt and tilorixan or its salt as active ingredients, and also includes pharmaceutically acceptable excipients or auxiliary ingredients. In the present disclosure, the dosage form of the drug preferably includes one or more of tablets, powders, granules, capsules, oral liquids and sustained-release preparations. The present disclosure does not specifically limit the amount of the withaferin A or its salt and tilorixan or its salt in the above dosage forms, and the amount of the drug in the conventional dosage form can be used. The present disclosure does not specifically limit the type and content of the excipients or auxiliary ingredients used in the above dosage forms, and conventional ones can be used.

[0064] According to an embodiment of the present disclosure, in the use of the pharmaceutical composition or combination drug of the present disclosure in the preparation of a medicament for preventing or treating neurodegenerative diseases, the pharmaceutical composition or combination drug contains about 50 mg to 200 mg of withaferin A or a salt thereof.

[0065] According to an embodiment of the present disclosure, in the use of the pharmaceutical composition or combination drug of the present disclosure in the preparation of a medicament for preventing or treating a neurodegenerative disease, the pharmaceutical composition or combination drug contains about 50 mg to 200 mg of tilorixen or a salt thereof.

[0066] According to an embodiment of the present disclosure, in the use of the pharmaceutical composition or combination drug of the present disclosure in the preparation of a medicament for preventing or treating neurodegenerative diseases, in the pharmaceutical composition or combination drug, the dosage ratio of withaferin A or its salt and tilorixen or its salt is about 2:1 to about 1:2, preferably about 1:1.

[0067] According to an embodiment of the present disclosure, in the use of the pharmaceutical composition or combination drug of the present disclosure in the preparation of a medicament for preventing or treating a neurodegenerative disease, the neurodegenerative disease is selected from Alzheimer's disease, Parkinson's disease, or Huntington's disease.

[0068] The present disclosure also provides use of withaferin A or a salt thereof in preparing a combined pharmaceutical composition for preventing or treating neurodegenerative diseases, wherein the combined pharmaceutical composition comprises withaferin A or a salt thereof and tilorixan or a salt thereof.

[0069] The present disclosure also provides use of tilorixan or a salt thereof in preparing a combination pharmaceutical composition for preventing or treating neurodegenerative diseases, wherein the combination pharmaceutical composition comprises withaferin A or a salt thereof and tilorixan or a salt thereof.

[0070] According to an embodiment of the present disclosure, in the use of withaferin A or its salt and tilorixan or its salt disclosed in the present disclosure in preparing a combination pharmaceutical composition for preventing or treating neurodegenerative diseases, the combination pharmaceutical composition comprises a therapeutically effective amount of withaferin A or its salt, a therapeutically effective amount of tilorixan or its salt, and a pharmaceutically acceptable carrier or excipient.

[0071] According to an embodiment of the present disclosure, in the use of withaferin A or its salt and tilorixan or its salt in the preparation of a combined pharmaceutical composition for preventing or treating neurodegenerative diseases, the pharmaceutical composition contains about 50 mg to 200 mg of withaferin A or its salt.

[0072] According to an embodiment of the present disclosure, in the use of withaferin A or its salt and tilorixan or its salt of the present disclosure in preparing a combined pharmaceutical composition for preventing or treating neurodegenerative diseases, the pharmaceutical composition contains about 50 mg to 200 mg of tilorixan or its salt.

[0073] According to an embodiment of the present disclosure, in the use of withaferin A or its salt and tilorexin or its salt of the present disclosure in preparing a combination pharmaceutical composition for preventing or treating neurodegenerative diseases, the dosage ratio of withaferin A or its salt and tilorexin or its salt in the pharmaceutical composition is about 2:1 to about 1:2, preferably about 1:1.

[0074] According to an embodiment of the present disclosure, in the use of withaferin A or its salt and tilorixan or its salt in the present disclosure in preparing a combination pharmaceutical composition for preventing or treating a neurodegenerative disease, the neurodegenerative disease is selected from Alzheimer's disease, Parkinson's disease, or Huntington's disease.

[0075] The present disclosure has no special limitation on the sources of withaferin A or its salt and tilorixan or its salt, and conventional commercial products can be used.

[0076] The present disclosure also provides the use of the pharmaceutical composition or combination drug described in the above technical solution in the preparation of a neuron-protective drug. In the present disclosure, different drug dosages are administered, and enhancement of object recognition ability is detected by a novel object recognition test, enhancement of spatial memory ability is detected by a novel location recognition test, or enhancement of working memory is detected by a T-maze test.

[0077] The present disclosure also provides a method for preventing or treating a neurodegenerative disease, comprising administering a therapeutically effective amount of withaferin A or a salt thereof to a subject in need thereof, and administering a therapeutically effective amount of tilorixan or a salt thereof to the subject.

[0078] According to an embodiment of the present disclosure, in the method for preventing or treating neurodegenerative diseases of the present disclosure, the administration of withaferin A or a salt thereof and tilorixan or a salt thereof are performed simultaneously or sequentially in any order.

[0079] According to an embodiment of the present disclosure, in the method for preventing or treating a neurodegenerative disease of the present disclosure, withaferin A or a salt thereof is administered in an amount of about 1 mg / kg to about 2 mg / kg, once every two days.

[0080] According to an embodiment of the present disclosure, in the method for preventing or treating a neurodegenerative disease of the present disclosure, tilorixen or a salt thereof is administered in an amount of about 1 mg / kg to about 2 mg / kg, once every two days.

[0081] According to an embodiment of the present disclosure, in the method for preventing or treating neurodegenerative diseases of the present disclosure, the administration ratio of withaferin A or its salt and tilorixan or its salt is about 2:1 to about 1:2, preferably about 1:1.

[0082] According to an embodiment of the present disclosure, in the method for preventing or treating a neurodegenerative disease of the present disclosure, the neurodegenerative disease is selected from Alzheimer's disease, Parkinson's disease, or Huntington's disease.

[0083] The present disclosure also provides the use of withaferin A or a salt thereof in enhancing the effect of tilorexin or a salt thereof in preventing or treating neurodegenerative diseases in a subject. Specifically, the use of withaferin A or a salt thereof in preparing a potentiator for tilorexin or a salt thereof in preventing or treating neurodegenerative diseases.

[0084] According to an embodiment of the present disclosure, the amount of withaferin A or its salt is as defined herein.

[0085] The present disclosure also provides the use of tilorixan or a salt thereof in enhancing the effect of withaferin A or a salt thereof in preventing or treating neurodegenerative diseases in a subject. Specifically, the use of tilorixan or a salt thereof in preparing a potentiator for withaferin A or a salt thereof in preventing or treating neurodegenerative diseases.

[0086] According to an embodiment of the present disclosure, the amount of tilorixen or its salt is as defined herein.

[0087] According to an embodiment of the present disclosure, the dosage ratio of withaferin A or its salt and tilorixan or its salt is as defined herein.

[0088] According to an embodiment of the present disclosure, the neurodegenerative disease is selected from Alzheimer's disease, Parkinson's disease, or Huntington's disease.

[0089] Before further describing the present disclosure, the following sections collect certain terms used in the specification, examples, and appended claims. The definitions listed herein should be read and understood by those skilled in the art in light of the remainder of this disclosure. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those of ordinary skill in the art to which this disclosure belongs.

[0090] definition

[0091] All numerical designations used herein, such as pH values, temperatures, times, concentrations, amounts, and molecular weights, including ranges, are approximate and, where appropriate, vary by increments of 0.1 or 1.0 (+) or (-). It will be understood that, although not always explicitly stated, all numerical designations may be preceded by the term "about."

[0092] The present disclosure is not limited to the specific systems, devices, and methods described, as these may vary. The terminology used in this specification is for the purpose of describing a particular version or embodiment only and is not intended to limit the scope. These aspects of the present disclosure may be embodied in many different forms; rather, these embodiments are provided so that this disclosure will be thorough and complete, and will fully convey its scope to those skilled in the art.

[0093] As used herein, singular forms include plural references unless the context clearly dictates otherwise. With respect to the use of substantially any plural and / or singular terms herein, those skilled in the art can translate from the plural to the singular and / or from the singular to the plural, depending on the context and / or application. For clarity, various singular / plural permutations may be explicitly set forth herein.

[0094] As will be understood by those skilled in the art, for any and all purposes, such as from the perspective of providing a written description, all ranges disclosed herein are intended to encompass every intermediate value between the upper and lower limits of the range, as well as any other stated or intermediate values ​​within the stated range. All ranges disclosed herein also include any and all possible subranges and combinations of subranges thereof. Any listed range can be easily identified as fully describing and capable of breaking down the same range into at least equal halves, thirds, quarters, fifths, tenths, etc. As a non-limiting example, each range discussed herein can be easily broken down into a lower third, a middle third, and an upper third, etc. As will also be understood by those skilled in the art, all language such as "up to," "at least," etc. includes the listed numbers and refers to ranges that can subsequently be broken down into subranges as discussed above. Finally, as will be understood by those skilled in the art, ranges include each individual member. Thus, for example, a group having 1-3 units refers to a group having 1, 2, or 3 units, as well as ranges having values ​​greater than or equal to 1 unit and less than or equal to 3 units. Similarly, a group having 1-5 cells refers to groups having 1, 2, 3, 4, or 5 cells, as well as ranges of values ​​greater than or equal to 1 cell and less than or equal to 5 cells, and so on.

[0095] Furthermore, even when a specific number is explicitly recited, one skilled in the art will recognize that such recitation should be interpreted as meaning at least the recited number (e.g., simply reciting "two statements" without other modifiers means at least two statements or two or more statements). Furthermore, in those instances where phrases similar to "at least one of A, B, and C, etc." are used, generally, such constructions are intended to mean the phrases as understood by those skilled in the art (e.g., "a system having at least one of A, B, and C" would include, but are not limited to, systems having A alone, B alone, C alone, A and B together, A and C together, B and C together, and / or A, B, and C together, etc.). In those instances where phrases similar to "at least one of A, B, or C, etc." are used, generally, such constructions are intended to mean the phrases as understood by those skilled in the art (e.g., "a system having at least one of A, B, or C" would include, but are not limited to, systems having A alone, B alone, C alone, A and B together, A and C together, B and C together, and / or A, B, and C together, etc.). Those skilled in the art will further understand that, whether in the specification, sample embodiments, or drawings, virtually any disjunctive word and / or phrase presenting two or more alternative terms should be understood to contemplate the possibility of including one of the terms, any of the terms, or both of the terms. For example, the phrase "A or B" will be understood to include the possibility of "A" or "B" or "A and B." Those skilled in the art will also understand that all language, such as "at most," "at least," "above," "below," etc., includes the recited number and refers to a range that can be subsequently subdivided into the sub-ranges discussed above.

[0096] As used herein, the term "about" refers to a change in a numerical value, which may occur, for example, by measurements or processing procedures in the real world, by errors in carelessness in these procedures, by differences in the production, source or purity of compositions or combination drugs or reagents, etc. Typically, as used herein, the term "about" means 1 / 10, such as ±10%, of a stated value or range of values ​​that is greater or less than the stated value. The term "about" also refers to changes that are considered to be equivalent by those skilled in the art, as long as these changes do not encompass known values ​​practiced by the prior art. Each value or range of values ​​preceded by the term "about" is also intended to include embodiments of the stated absolute value or range of values. Regardless of whether modified by the term "about", the quantitative values ​​narrated in this disclosure include the equivalents of the stated values, such as, the numerical values ​​of these values ​​may change, but those skilled in the art will recognize that these changes are equivalents. Where the context of this disclosure indicates otherwise or is inconsistent with this interpretation, the interpretation as described above may be modified, as will be apparent to those skilled in the art.

[0097] As used herein, "optional" or "optionally" means that the subsequently described event or circumstance can or cannot occur, and that the description includes instances where said event or circumstance occurs and instances where it does not.

[0098] Those skilled in the art will understand that, in general, the terms used herein are generally intended to be "open" terms (e.g., the term "including" should be interpreted as "including but not limited to," the term "having" should be interpreted as "having at least," the term "comprising" should be interpreted as "including but not limited to," etc.). In addition, the transitional term "comprising" (which is synonymous with "including," "containing," or "characterized by") is inclusive or open-ended and does not exclude additional, unrecited elements or method steps. Although various compositions, methods, and apparatus are described in terms of "including" various components or steps (interpreted as "including but not limited to"), the compositions, methods, and apparatus may also "consist essentially of" or "consist of" the various components and steps, and such terms should be interpreted as defining a substantially closed group of members. In contrast, the transitional phrase "consisting of" excludes any element, step, or ingredient not specified in the claim. The transitional phrase "consisting essentially of" limits the scope of the claim to the specified materials or steps "and those materials or steps that do not materially affect the basic and novel characteristics of the claimed invention."

[0099] The term "combination therapy" or "combination drug" or "drug combination" means the administration of two or more therapeutic agents to treat the therapeutic conditions or disorders described in this disclosure. Such administration includes co-administration of these therapeutic agents in a substantially simultaneous manner, for example, in a single capsule having a fixed ratio of active ingredients or in multiple separate capsules for each active ingredient. In addition, such administration also includes the use of each type of therapeutic agent in a sequential manner. In either case, the treatment regimen will provide the beneficial effects of the drug combination in treating the conditions or disorders described herein.

[0100] Those skilled in the art should understand that when the pharmaceutical composition of the present disclosure contains i) withaferin A or a salt thereof and ii) tilorixen or a salt thereof, it means that it may include but is not limited to the following schemes: (1) the pharmaceutical composition of the present disclosure contains an agent that contains both withaferin A or a salt thereof and tilorixen or a salt thereof as active substances; (2) the pharmaceutical composition of the present disclosure contains a first agent containing withaferin A and also contains a second agent containing tilorixen.

[0101] As used herein, the term "disease" is generally synonymous with the terms "disorder," "syndrome," and "condition" (as in medical conditions) and are used interchangeably because they all reflect an abnormal condition of the human or animal body or one of its parts that impairs normal function, is usually manifested by distinctive signs and symptoms, and results in a decrease in the lifespan or quality of life of the human or animal.

[0102] As used herein, the term "treatment" refers to both therapeutic treatment and preventative measures, wherein the purpose is to prevent or slow down (mitigate) an undesirable physiological condition, disorder, or disease, or to obtain a beneficial or desired clinical outcome. For the purposes of this disclosure, beneficial or desired clinical outcomes include, but are not limited to, alleviation of symptoms, alleviation of the extent of the condition, disorder, or disease, stabilization of the condition, disorder, or disease (i.e., no worsening), delayed onset or slowed progression of the condition, disorder, or disease, improvement of the condition, disorder, or disease, and detectable or undetectable relief (whether partial or complete) or promotion or improvement of the condition, disorder, or disease. Treatment includes eliciting a clinically significant response without excessive levels of side effects. Treatment also includes extending survival compared to the expected survival if not receiving treatment. Treatment may also be proactive in nature, i.e., it may include preventing the disease. Prevention of a disease may involve complete protection from the disease, such as in the case of preventing infection with a pathogen, or may involve preventing the progression of the disease. For example, prevention of a disease may not mean the complete loss of any effect associated with the disease at any level, but may mean preventing the symptoms of the disease from reaching a clinically significant or detectable level. Prevention of a disease may also mean preventing the disease from developing to a later stage of the disease. As used herein, "treatment" covers the treatment of diseases (primarily neurodegenerative diseases) in mammals, especially humans, including: (a) preventing the occurrence of diseases (e.g., preventing neurodegenerative diseases) or symptoms in individuals who are susceptible to the disease but have not yet been diagnosed with the disease; (b) inhibiting the disease, such as arresting the progression of the disease; or (c) alleviating the disease, such as alleviating the symptoms associated with the disease. "Treatment" as used herein covers any medication that administers a drug or compound to an individual to treat, cure, alleviate, improve, mitigate or inhibit the individual's disease, including but not limited to administering a drug containing lipoic acid and inca inchi oil as described herein to an individual in need.

[0103] As used herein, the term "drug" means an agent used to treat, combat, ameliorate, prevent, or improve an unwanted condition or disease in a subject.

[0104] The term "pharmaceutically acceptable" refers to compositions or combinations of drugs that are suitable for use in contact with the tissues of subjects without excessive toxicity, irritation and allergic response, commensurate with a reasonable benefit / risk ratio, and effective for their intended purpose.

[0105] The phrase "therapeutically effective amount" is intended to qualify the amount of active ingredient used in the treatment of a disease or disorder or in the achievement of a clinical endpoint.

[0106] The term "effective amount" is used herein to refer to an amount of a compound that, when administered to a subject, is suitable for achieving the purpose of the compound, including imaging of subject tissue, diagnosing a condition in a subject, and / or monitoring a subject for symptoms or disorders. The actual amount comprising an "effective amount" will vary depending on a variety of circumstances, including but not limited to the severity of the disease, the size and health of the subject, the imaging modality, the diagnostic modality, the monitoring modality, and the route of administration. A skilled medical practitioner can readily determine the appropriate amount using methods known in the medical arts.

[0107] The term "therapeutically effective amount" is used herein to refer to an amount of a compound that, when administered to a subject, is capable of alleviating the symptoms of a disorder in the subject or enhancing the texture, appearance, color, feel, or hydration of the intended tissue treatment area. The actual amount comprising a "therapeutically effective amount" will vary depending on a variety of circumstances, including, but not limited to, the severity of the disease, the size and health of the subject, and the route of administration. A skilled medical practitioner can readily determine such an appropriate amount using methods known in the medical art.

[0108] As used herein, the terms "excipient" and "pharmaceutically acceptable excipient" are generally synonymous with the terms "carrier," "pharmaceutically acceptable carrier," "diluent," and "pharmaceutically acceptable diluent," and are used interchangeably.

[0109] As used herein, the term "carrier" includes carriers, excipients, and diluents, and refers to materials, compositions, or carriers that participate in carrying or transporting drugs, cosmetics, or other medicaments across tissue layers (e.g., the stratum corneum or the stratum spinosum), such as liquid or solid fillers, diluents, excipients, solvents, or encapsulating materials. The pharmaceutical composition or combination of the compound may also include a suitable solid or gel phase carrier or excipient. Examples of such carriers or excipients include, but are not limited to, calcium carbonate, calcium phosphate, various sugars, starches, cellulose derivatives, gelatin, and polymers such as polyethylene glycol.

[0110] As used herein, "salt" includes acid addition salts or base addition salts of the compound.

[0111] Suitable acid addition salts are formed from acids which form non-toxic salts. Examples include acetate, adipate, aspartate, benzoate, benzenesulfonate, bicarbonate / carbonate, bisulfate / sulfate, borate, camphorsulfonate, citrate, cyclamate, edisylate, ethanesulfonate, formate, fumarate, gluconate, glucuronate, hexafluorophosphate, hibenzate, hydrochloride / chloride, hydrobromide / bromide, hydroiodide / iodide, isethionate, lactate, malate, maleate, malonate, methanesulfonate, methylsulfate, naphthoate, 2-naphthalenesulfonate, nicotinate, nitrate, orotate, oxalate, palmitate, pamoate, phosphate / hydrogenphosphate / dihydrogenphosphate, pyroglutamate, aldarate, stearate, succinate, tannate, tartrate, tosylate, trifluoroacetate, and xinofoate.

[0112] Suitable base addition salts are formed from bases which form non-toxic salts. Examples include aluminum, arginine, benzathine, calcium, choline, diethylamine, diethanolamine, glycine, lysine, magnesium, meglumine, ethanolamine, potassium, sodium, tromethamine, and zinc salts.

[0113] The terms "subject" and "patient" are interchangeable and can mean any living organism that can be treated with the compositions or combinations of the present disclosure. Thus, the terms "subject" and "patient" can include, but are not limited to, any non-human mammal, primate, or human. In some embodiments, the "subject" or "patient" is a mammal, such as a mouse, rat, other rodent, rabbit, dog, cat, pig, cattle, sheep, horse, primate, or human. In some embodiments, the subject or patient is an adult, child, or infant. In some embodiments, the subject or subject is a human.

[0114] Where the present disclosure refers to the term "doctor" and other terms for various medical professionals in terms of a particular job title or role, nothing in the present disclosure is intended to be limited to that particular job title or role. A doctor or medical professional may include any physician, nurse, medical professional, or technician. Unless expressly defined otherwise, any of these terms or job titles may be used interchangeably by users of the systems disclosed herein. For example, in some embodiments, reference to a doctor may also apply to a technician, nurse, or other healthcare provider.

[0115] As used herein, the term "administering" refers to the administration of a compound (also referred to as an agent of interest), a combination drug, a pharmaceutically acceptable salt of the compound (agent of interest), a pharmaceutically acceptable salt of the combination drug, or a composition thereof, directly to the subject by the subject or a healthcare provider.

[0116] As used herein, the term "diagnosis" refers to the process of identifying the presence and / or nature of a disease, disorder, or other physiological state in a subject based on its characteristics, signs, and symptoms. Diagnosis can include statements or conclusions related to a disease, disorder, or other physiological state in a subject based on such a process.

[0117] The term "inhibit" includes administering a composition or combination of the present disclosure to prevent the onset of symptoms, alleviate symptoms, reduce symptoms, slow or reduce the progression of a disease and / or its symptoms, or eliminate a disease, condition, or disorder.

[0118] In some embodiments, the methods, compositions, or combinations disclosed herein can be used for subjects who require such examination, diagnosis, monitoring, and / or treatment, which can also be referred to as "in need thereof." As used herein, the phrase "in need thereof" means that the subject has been identified as requiring a particular method or treatment, or has been identified as having a condition, and the method (e.g., tissue imaging, condition diagnosis, condition monitoring) or treatment has been used for the subject for that specific purpose.

[0119] The compositions of the present disclosure or the combination drug can be used in a conventional manner by any route in which they have activity. Administration can be systemic, topical or oral. For example, administration can be but is not limited to parenteral, subcutaneous, intravenous, intramuscular, intraperitoneal, transdermal, oral, cheek or eye route, or intravaginal, by inhalation, by reservoir injection or by implant. Therefore, the mode of administration (alone or in combination with other drugs) can be but is not limited to sublingual, injection (including short-acting, reservoir, implant and granular form of subcutaneous or intramuscular injection), topical (including nasal spray, ointment or cream, for example, applied to the skin) and / or by using vaginal cream, suppository, pessary, vaginal ring, rectal suppository, intrauterine device and transdermal form such as patch and cream.

[0120] The specific mode of administration will depend on the indication or purpose. The selection of specific route of administration and dosage regimen will be adjusted or titrated by the clinician according to methods known to the clinician to obtain the best clinical response. The amount of the compound to be administered is an effective amount. The dosage to be administered will depend on the characteristics of the subject being treated, such as the specific animal being treated, age, body weight, health, type and frequency of treatment (if any) simultaneously, and can be easily determined by those skilled in the art (e.g., by a clinician).

[0121] For oral administration, by combining withaferin A and tilorixen with pharmaceutically acceptable carriers known in the art, compositions or combination drugs can be easily formulated by the methods herein. These carriers enable the compounds of the present disclosure to be formulated into tablets, pills, dragees, capsules, liquids, gels, syrups, slurries, suspensions, etc. for oral ingestion by subjects to be imaged, diagnosed and / or treated. Pharmaceutical preparations for oral use can be obtained as follows: adding a solid excipient, optionally grinding the resulting mixture, and treating the mixture of particles after adding a suitable adjuvant (if necessary) to obtain tablets or dragee cores. Suitable excipients include, but are not limited to, fillers, such as sugars, including but not limited to lactose, sucrose, mannitol, and sorbitol; cellulose preparations, such as, but not limited to, corn starch, wheat starch, rice starch, potato starch, gelatin, tragacanth gum, methylcellulose, hydroxypropyl methylcellulose, sodium carboxymethylcellulose, and polyvinylpyrrolidone (PVP). If desired, disintegrating agents may be added, such as, but not limited to, the cross-linked polyvinyl pyrrolidone, agar, or alginic acid or a salt thereof such as sodium alginate.

[0122] Orally usable pharmaceutical compositions or combinations include, but are not limited to, push-fit capsules made of gelatin, and soft sealed capsules made of gelatin and a plasticizer (such as glycerol or sorbitol). Push-fit capsules can contain active ingredients mixed with fillers (such as lactose), binders (such as starch) and / or lubricants (such as talc or magnesium stearate) and optional stabilizers. In soft capsules, the active compound can be dissolved or suspended in a suitable liquid such as a fatty oil, liquid paraffin or liquid polyethylene glycol. In addition, stabilizers can be added. The capsule can also be coated with an additional layer to protect the contents and / or delay the release of the contents through one or more digestive stages. For example, the capsule or other carrier can include an enteric coating (such as formed from a polymer) to prevent dissolution or disintegration in the gastric environment. All compositions or combinations for oral administration should have a dosage suitable for such administration.

[0123] For injection administration, the porous microparticles are usually combined with one or more pharmaceutically acceptable bases for injection (e.g., suspended therein). The pharmaceutically acceptable base can be any known aqueous or non-aqueous base in the prior art. Examples of aqueous bases include physiological saline solutions, sugars such as dextrose or mannitol solutions and pharmaceutically acceptable buffers, and examples of non-aqueous bases include fixed vegetable oils, glycerol, polyethylene glycol, alcohols, and ethyl oleate. The base can also include antimicrobial preservatives, antioxidants, tonicity agents, buffers, stabilizers, or other components.

[0124] Compositions comprising herein can be administered by injection in a pharmaceutically acceptable carrier and be applied to a patient for local, regional, or systemic delivery of a pharmaceutical substance. Injection is generally performed with a conventional syringe and needle, catheter, or the like. In other embodiments, more complex delivery systems can be used, such as needleless syringes, to inject the preparation. The pharmaceutical preparation can be injected into almost any organ or region of the body, including intravenous, intramuscular, intradermal, subcutaneous, intraarticular, intrasynovial, intraosseous, intraspinal, intrathecal, intraarticular, or intracardial administration. In some other embodiments, the preparation is suitable for intracranial, intralesional, or intratumoral administration.

[0125] The term "tissue" refers to any collection of similar specialized cells united to perform a specific function.

[0126] In this article, Withaferin A, with the molecular formula C 28 H 38 O6 is a purified product of withanolide from the root extract of South African somniferum. It is a natural steroid compound. Withaferin A is easily soluble in organic solvents such as dimethyl sulfoxide, methanol, and ethanol, but difficult to dissolve in water. Withaferin A can pass through the blood-brain barrier but is almost insoluble in water. Using organic solvents such as dimethyl sulfoxide as a solvent will inevitably inhibit cell activity. In this article, tilorixan, with the molecular formula C 17 H 26 ClNO is a selective histamine 3 (H3) receptor antagonist. H3 receptors are primarily located in the cerebral cortex, hypothalamus, hippocampus, and basal ganglia. Blockade of histamine autoreceptors by tilorixen increases histamine concentrations and histaminergic activity in the brain.

[0127] According to the embodiments of the present disclosure, the drugs or pharmaceutical compositions or combination drugs of the present disclosure can be used in combination with conventional treatment methods and / or therapies, or can be used separately from conventional treatment methods and / or therapies. When the drugs or pharmaceutical compositions or combination drugs of the present disclosure are administered in combination therapy with other drugs, they can be administered to an individual sequentially or simultaneously. Alternatively, the pharmaceutical composition or combination drug of the present disclosure can comprise a combination of withaferin A and tilorixan of the present disclosure, a pharmaceutically acceptable carrier or a pharmaceutically acceptable excipient, and other therapeutic or prophylactic drugs known in the art.

[0128] The present disclosure proposes a drug combination. According to an embodiment of the present disclosure, the pharmaceutical composition or combination drug is used to prevent or treat neurodegenerative diseases, preferably for preventing or treating Alzheimer's disease and / or Parkinson's disease, and more preferably for preventing or treating Alzheimer's disease. The drug combination includes: withaferin A or a pharmaceutically acceptable salt thereof as a first active agent, and tilorixan or a pharmaceutically acceptable salt thereof as a second active agent. The drug combination according to the embodiments of the present disclosure has good effect in preventing and / or treating neurodegenerative diseases with few side effects.

[0129] Neurodegenerative diseases within the meaning of the present disclosure include, but are not limited to, multiple sclerosis (MS), Parkinson's disease (PD), Huntington's disease (HD), amyotrophic lateral sclerosis (ALS), stroke and dementia, the latter including, but not limited to, Alzheimer's disease (AD), vascular dementia, frontotemporal dementia, semantic dementia and Lewy body dementia. Preferred neurodegenerative diseases are multiple sclerosis, Alzheimer's disease, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis and stroke. In some embodiments, the neurodegenerative disease is a neurodegenerative disease characterized by the formation of amyloid plaques in the brain. In a specific preferred embodiment, the neurodegenerative disease is Alzheimer's disease or Parkinson's disease. Preferably, the neurodegenerative disease is Alzheimer's disease. The inventors have found that the composition or combination of withaferin A and tilorexin is more effective in treating Alzheimer's disease.

[0130] According to an embodiment of the present disclosure, the neural cells are human neuroblastoma cells. The inventors have discovered that a mixture of lipoic acid and inca inchi oil is more effective in increasing the survival rate of human neuroblastoma cells, reducing apoptosis in human neuroblastoma cells, protecting the mitochondrial membrane potential of human neuroblastoma cells, and protecting the energy metabolism level of human neuroblastoma cells.

[0131] By reserving the right to exclude or exclude any individual member of any such group (including any subrange or combination of subranges in such group) by limitation, which can be claimed by scope or any similar means, less than the entire measure of the present disclosure may be claimed for any reason. In addition, by reserving the right to exclude or exclude any individual substituent, structure or group thereof, or any member of the claimed group by limitation, less than the entire measure of the present disclosure may be claimed for any reason. Various patents, patent applications and publications are cited throughout this disclosure. The disclosures of these patents, patent applications and publications are incorporated into this disclosure in their entirety by reference to more fully describe the state of the art known to those skilled in the art as of the date of this disclosure. In the event of any inconsistency between the cited patents, patent applications and publications and the present disclosure, the present disclosure shall prevail.

[0132] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. Nothing in this disclosure should be construed as an admission that the embodiments described in this disclosure are not entitled to antedate such disclosure by virtue of prior invention.

[0133] Also provided are embodiments in which any embodiment herein can be combined with any one or more of the other embodiments, unless otherwise stated, and provided that the combinations are not mutually exclusive.

[0134] The technical solutions provided by the present disclosure are described in detail below in conjunction with the embodiments, but they should not be understood as limiting the scope of protection of the present disclosure.

[0135] Materials and methods

[0136] 1. Experimental Animals

[0137] The mice used in the following examples were: 6-8 month old female C57 5XFAD mice, purchased from Yangzhou Youdu Biotechnology Co., Ltd. 5XFAD mice are a mouse model of Alzheimer's disease (AD) that accumulates Aβ plaques. 5XFAD mice overexpress the amino acid isoform of human amyloid precursor protein (APP695) under the control of the mouse thy-1 promoter. In addition, the human presenilin-1 (PS1) gene is also expressed under the control of the mouse thy-1 promoter in mice carrying the M14 6L and L28 6V mutations. 5XFAD mice exhibited working memory impairment in alternating experiments and also developed age-dependent motor dysfunction. Only female mice were used in the current study.

[0138] 2. C57 5XFAD Mouse Housing Conditions

[0139] The ambient temperature was 22 ± 0.5°C, with a 12-hour light / dark cycle. All experimental data are expressed as mean ± standard error. *P < 0.05, **P < 0.01, ***P < 0.001, n = 6 / 8. P < 0.05 was considered to indicate a significant difference between groups.

[0140] 3. Other Reagents

[0141] WITHAFERIN A: purchased from TargetMol, mass 25 mg

[0142] Pitolisant hydrochloride: purchased from Yuanye Biotechnology, mass 100 mg

[0143] DMSO: purchased from Yuanye Biotechnology, volume 100 mL

[0144] Example

[0145] Example 1. Preparation of a pharmaceutical composition or combination drug containing withaferin A and tilorixan

[0146] (1) Tablet preparation

[0147] The raw and auxiliary materials were passed through an 80-mesh sieve for later use. The prescribed amount of active ingredients, microcrystalline cellulose, lactose, and povidone K30 were weighed and added to a high-speed mixer. The mixture was stirred at a low speed to mix evenly. An appropriate amount of purified water was added, the mixture was stirred at a low speed, and the mixture was granulated by high-speed cutting. The wet granules were dried at 60°C for 3 h, sieved through a 24-mesh sieve, and the prescribed amount of sodium carboxymethyl starch, silicon dioxide, and magnesium stearate were added. The mixture was mixed and tableted using a rotary tablet press.

[0148] The tablets can also be further prepared as injection solutions.

[0149] (2) Capsule preparation

[0150] The raw and auxiliary materials were sieved through an 80-mesh sieve for later use. The prescribed amount of active ingredients, lactose, starch, and povidone K30 were weighed and added to a high-speed mixer. The mixture was stirred at a low speed to mix evenly. An appropriate amount of purified water was added, the mixture was stirred at a low speed, and the mixture was granulated by high-speed cutting. The wet granules were dried at 60°C for 3h, sieved through a 24-mesh sieve, and the prescribed amount of silicon dioxide and magnesium stearate were added. The mixture was mixed and filled into capsules using a capsule filling machine.

[0151] (3) Preparation of injections

[0152] a. Preparation of 2 mg / kg Withaferin A injection

[0153] Dissolve 20 mg of Withaferin A micropowder in 20 mL of DMSO buffer to prepare a 1 mg / mL Withaferin A active ingredient injection. Before administration, weigh the mouse and determine the volume of injection based on the administered dose.

[0154] b. Preparation of 1mg / kg Withaferin A injection

[0155] Dissolve 20 mg of Withaferin A micropowder in 40 mL of DMSO buffer to prepare a 0.5 mg / mL Withaferin A active ingredient injection. Before administration, weigh the mouse and determine the volume of injection based on the administered dose.

[0156] c. Preparation of 2mg / kg Tilorisen Injection

[0157] Dissolve 20 mg of tilorixen micropowder in 40 mL of 0.9% sodium chloride buffer to prepare a 0.5 mg / mL tilorixen saline injection. Before administration, weigh the mouse and determine the volume of injection based on the administered dose.

[0158] Example 2. Disease model establishment

[0159] Alzheimer's disease (AD) model establishment: 6- to 8-month-old female C57 5XFAD mice were purchased and, after adaptive feeding for 1 week, mice weighing (25±3) g were randomly selected for the experiment; alternatively, 6-month-old male heterozygous C57 5XFAD mice were bred with female C57 mice and the offspring were genetically identified, and offspring mice weighing (25±3) g were selected for the experiment.

[0160] At this time, 24 6-8 month old female C57 5XFAD mice were selected as the experimental group, 6 6-8 month old female C57 5XFAD mice were selected as the control group (DMSO was selected as the injection solvent for the control group), and 6 6-8 month old female C57 mice were selected as the same group control (WT).

[0161] The experimental mice were divided into four groups:

[0162] The first group (AD+2W) received intraperitoneal injection of 1 μg / μL withaferin A 0.05 mL (2 mg / kg) every two days;

[0163] The second group (AD+2P) received intraperitoneal injection of 0.1 mL (2 mg / kg) of tilorixen at a concentration of 0.5 μg / μL every two days;

[0164] The third group (AD+1W+1P) received intraperitoneal injections of 0.5 μg / μL withaferin A 0.05 mL (1 mg / kg) and 0.25 μg / μL tilorexan 0.1 mL (1 mg / kg) every two days;

[0165] The fourth group (AD+2W+2P) was intraperitoneally injected with 0.05 mL (2 mg / kg) of withaferin A at a concentration of 1 μg / μL and 0.1 mL (2 mg / kg) of tilorixen at a concentration of 0.5 μg / μL every two days.

[0166] The mice in the control group (AD+DMSO) were intraperitoneally injected with 0.05 mL of the same volume of control solvent every two days.

[0167] WT mice (C57 normal mice) were not treated.

[0168] *W refers to the active ingredient withaferin A, and P refers to the active ingredient that replaces Loli.

[0169] Example 3. Behavioral Experiment

[0170] After intraperitoneal injection of the experimental and control groups, the changes in behavioral indicators of each group were detected. All temperatures are expressed in °C, and all reactions were performed at room temperature (RT) unless otherwise stated.

[0171] Behavioral Indicator Testing: After establishing the Alzheimer's disease model, drug injections were performed and behavioral indicators of mice were tested in a quiet, appropriate environment. Each experiment was repeated multiple times to eliminate the interference of other factors on behavioral indicators.

[0172] No animals died after injection or during behavioral testing.

[0173] 1. New Object Recognition Experiment

[0174] (1) Construct a test room. Select two connected rooms and test the behavioral indicators of mice in a dark room with only one side illuminated.

[0175] (2) Prepare a white open field with a length, width and height of 40 cm and a wall thickness of 5 mm.

[0176] (3) Move the mice to the testing room and habituate them for 1 hour.

[0177] (4) The mouse was placed facing forward in the center of the open field and habituated for 10 min. The crawling distance in the open field was recorded.

[0178] (5) Construct a new object recognition learning environment and place two identical objects on the same side of the open field.

[0179] (6) Place the mouse in the center of the open field with its head facing forward and facing away from the object, and allow it to learn for 10 minutes.

[0180] (7) Remove the mouse and place it in its home cage for 10 min.

[0181] (8) Construct a new object recognition test environment and replace the objects on one side of the open field.

[0182] (9) Place the mouse in the center of the open field with its head facing forward and facing away from the object for 10 minutes.

[0183] (10) The same batch of mice were tested one week after administration, and the results are shown in Table 1 and Figure 1. Figure 1 shows the results of the analysis, which confirms that withaferin A and tilorixan can improve the object recognition ability of mice, and that the combined administration has the effect of 1+1>2.

[0184] Table 1. Object recognition index (%)

[0185] 2. New location recognition experiment

[0186] (1) Construct a test room. Select two connected rooms and test the behavioral indicators of mice in a dark room with only one side illuminated.

[0187] (2) Prepare a white open field with a length, width and height of 40 cm and a wall thickness of 5 mm.

[0188] (3) Move the mice to the testing room and habituate them for 1 hour.

[0189] (4) The mouse was placed facing forward in the center of the open field and habituated for 10 min. The crawling distance in the open field was recorded.

[0190] (5) Construct a new location recognition learning environment and place two identical objects on the same side of the open field.

[0191] (6) Place the mouse in the center of the open field with its head facing forward and facing away from the object, and allow it to learn for 10 minutes.

[0192] (7) Remove the mouse and place it in its home cage for 10 min.

[0193] (8) Construct a new location recognition test environment and place the object on one side of the open field on the opposite side of the open field.

[0194] (9) Place the mouse in the center of an open field with its head facing the opposite side and test for 10 minutes.

[0195] (10) The same batch of mice were tested one week after administration, and the results are shown in Table 2 and Figure 2. Figure 2 shows the results of the analysis, which confirms that withaferin A and tilorixan can improve the location recognition ability of mice and have a combined administration effect of 1+1>2.

[0196] Table 2. Position recognition index (%)

[0197] 3. T-maze test

[0198] (1) House mice individually and weigh them. Ensure adequate food and deprive them of water for 36 hours.

[0199] (2) Weigh the mice after 36 hours.

[0200] (3) Place two identical rubber trays at the same position on the two arms of the T-maze and inject 50 μL of water into one side.

[0201] (4) Place the mouse at the neck of the T-maze and allow it to drink water and habituate.

[0202] (5) Inject 50 μL of water into the contralateral rubber tray and repeat the experiment 10 times.

[0203] (6) Construct a T-maze experimental learning environment, inject 50uL of water into the trays of the two arms of the T-maze, and block one arm.

[0204] (7) Place the mouse at the neck of the T-maze and allow it to drink water and explore one arm.

[0205] (8) Remove the mouse and place it in the neck of the T-maze again for 1.5 minutes.

[0206] (9) Remove the barrier and allow the mouse to explore freely. If the mouse chooses the ipsilateral arm, it will be punished by being forced to stay still for 30 seconds and will not be able to drink water. If the mouse chooses the contralateral arm, it will be allowed to drink water.

[0207] (10) Four weeks after intraperitoneal injection, the same group of mice were tested 10 times per day for three consecutive days. Each five tests constituted a session, for a total of six sessions, with the first five sessions serving as the learning phase and the sixth session as the testing phase.

[0208] The results are shown in Table 3 and Figures 3 to 6. Figures 3 to 6 show the results of the analysis, which confirm that withaferin A and tilorixen can improve the location recognition ability of mice and have a combined administration effect of 1+1>2.

[0209] Table 3. Correct selection rate (%)

[0210] Finally, the various technical solutions involved in this disclosure are summarized again and listed below:

[0211] Item 1. A pharmaceutical composition comprising i) withaferin A or a salt thereof; and ii) tilorixan or a salt thereof.

[0212] Item 2. The pharmaceutical composition according to Item 1, comprising a therapeutically effective amount of withaferin A or a salt thereof, a therapeutically effective amount of tilorixan or a salt thereof, and a pharmaceutically acceptable carrier or excipient.

[0213] Item 3. The pharmaceutical composition according to Item 1 or 2, comprising about 1 mg to 4000 mg of withaferin A or a salt thereof.

[0214] Item 4. The pharmaceutical composition according to any one of Items 1 to 3, comprising about 1 mg to 4000 mg of tilorixen or a salt thereof.

[0215] Item 5. The pharmaceutical composition according to any one of Items 1 to 4, wherein the dosage ratio of withaferin A or its salt and tilorixan or its salt is about 10:1 to about 1:10, preferably about 1:1 or 1:2.

[0216] Item 6. The pharmaceutical composition according to any one of Items 1 to 5, which is used for preventing or treating a neurodegenerative disease.

[0217] Item 7. The pharmaceutical composition according to Item 6, wherein the neurodegenerative disease is selected from Alzheimer's disease, Parkinson's disease, or Huntington's disease.

[0218] Item 8. Use of a pharmaceutical composition in the preparation of a medicament for preventing or treating a neurodegenerative disease, wherein the pharmaceutical composition comprises i) withaferin A or a salt thereof; and ii) tilorixan or a salt thereof.

[0219] Item 9. The use according to Item 8, wherein the pharmaceutical composition comprises a therapeutically effective amount of withaferin A or a salt thereof, a therapeutically effective amount of tilorixan or a salt thereof, and a pharmaceutically acceptable carrier or excipient.

[0220] Item 10. The use according to Item 8 or 9, wherein the pharmaceutical composition comprises about 1 mg to 4000 mg of withaferin A or a salt thereof.

[0221] Item 11. The use according to any one of Items 8 to 10, wherein the pharmaceutical composition comprises about 1 mg to 4000 mg of tilorixen or a salt thereof.

[0222] Item 12. The use according to any one of Items 8 to 11, wherein in the pharmaceutical composition, the dosage ratio of withaferin A or its salt and tilorixan or its salt is about 10:1 to about 1:10, preferably about 1:1 or 1:2.

[0223] Item 13. The use according to any one of Items 8 to 12, wherein the neurodegenerative disease is selected from Alzheimer's disease, Parkinson's disease, or Huntington's disease.

[0224] Item 14. Use of withaferin A or a salt thereof in the preparation of a combined pharmaceutical composition for preventing or treating a neurodegenerative disease, the combined pharmaceutical composition comprising i) withaferin A or a salt thereof; and ii) tilorixan or a salt thereof.

[0225] Item 15. Use of tilorixan or a salt thereof in the preparation of a combined pharmaceutical composition for preventing or treating a neurodegenerative disease, the combined pharmaceutical composition comprising i) withaferin A or a salt thereof; and ii) tilorixan or a salt thereof.

[0226] Item 16. The use according to Item 14 or 15, wherein the combination composition comprises a therapeutically effective amount of withaferin A or a salt thereof, a therapeutically effective amount of tilorixan or a salt thereof, and a pharmaceutically acceptable carrier or excipient.

[0227] Item 17. The use according to any one of Items 14 to 16, wherein the pharmaceutical composition comprises about 1 mg to 4000 mg of withaferin A or a salt thereof.

[0228] Item 18. The use according to any one of Items 14 to 17, wherein the pharmaceutical composition comprises about 1 mg to 4000 mg of tilorixen or a salt thereof.

[0229] Item 19. The use according to any one of Items 14 to 18, wherein in the pharmaceutical composition, the dosage ratio of withaferin A and tilorixen is about 10:1 to about 1:10, preferably about 1:1 or about 1:2.

[0230] Item 20. The use according to any one of Items 14 to 19, wherein the neurodegenerative disease is selected from Alzheimer's disease, Parkinson's disease, or Huntington's disease.

[0231] Item 21. A method for preventing or treating a neurodegenerative disease, comprising administering a therapeutically effective amount of withaferin A or a salt thereof to a subject in need thereof, and administering a therapeutically effective amount of tilorixan or a salt thereof to the subject.

[0232] Item 22. The method according to Item 21, wherein the administration of withaferin A or a salt thereof and tilorixan or a salt thereof is performed simultaneously or sequentially in any order.

[0233] Item 23. The method according to Item 21 or 22, wherein withaferin A or a salt thereof is administered in an amount of about 0.02-40 mg / kg, preferably about 2-4 mg / kg, daily or once every two days.

[0234] Item 24. The method according to any one of Items 21 to 23, wherein tilorixen or a salt thereof is administered in an amount of about 0.02-40 mg / kg, preferably about 2-10 mg / kg, daily or once every two days.

[0235] Item 25. The method according to any one of Items 21 to 24, wherein the administered dosages of withaferin A or its salt and tilorixan or its salt are in a ratio of about 10:1 to about 1:10, preferably about 1:1 or 1:2.

[0236] Item 26. The method according to any one of Items 21 to 25, wherein the neurodegenerative disease is selected from Alzheimer's disease, Parkinson's disease, or Huntington's disease.

[0237] Item 27. Use of withaferin A or a salt thereof for enhancing the effect of tilorixen or a salt thereof in preventing or treating a neurodegenerative disease in a subject.

[0238] Item 28: Use of tilorixan or a salt thereof for enhancing the effect of withaferin A or a salt thereof in preventing or treating a neurodegenerative disease in a subject.

[0239] Item 29. Use of withaferin A or its salt in the preparation of a synergist for tilorixen or its salt for preventing or treating neurodegenerative diseases.

[0240] Item 30. Use of tilorixan or a salt thereof in the preparation of a synergist for withaferin A or a salt thereof for preventing or treating neurodegenerative diseases.

[0241] Item 31. The use according to Item 27 or 29, wherein the amount of withaferin A or a salt thereof is about 1 mg to 4000 mg.

[0242] Item 32. The use according to Item 28 or 30, wherein the amount of tilorixen or a salt thereof is about 1 mg to 4000 mg.

[0243] Item 33. The use according to any one of Items 27 to 30, wherein the dosage ratio of withaferin A or a salt thereof and tilorixan or a salt thereof is from about 10:1 to about 1:10, preferably about 1:1 or about 1:2.

[0244] Item 34. Use according to any one of Items 27 to 33, wherein the neurodegenerative disease is selected from Alzheimer's disease, Parkinson's disease, or Huntington's disease.

[0245] Item 35. The pharmaceutical composition according to item 6 or 7, the use according to any one of items 8 to 19 or any one of claims 27 to 33, the method according to any one of items 21 to 26, wherein the subject is a mammal, preferably a human subject.

[0246]

[00106] Methods described herein can be performed in any order that is logically possible, except in the specific order disclosed.

[0247] The representative examples are intended to help illustrate the present disclosure and are not intended to, and should not be construed as, limiting the scope of the present disclosure. Indeed, various modifications of the present disclosure and numerous other embodiments thereof, in addition to those shown and described herein, will become apparent to those skilled in the art, including the examples and the scientific and patent literature references cited herein. The examples contain important additional information, illustration, and guidance that can be employed in the practice of the present disclosure in its various embodiments and equivalents.

Claims

1. A pharmaceutical composition comprising i) withaferin A or a salt thereof; and ii) tilorixan or a salt thereof.

2. The pharmaceutical composition according to claim 1, comprising a therapeutically effective amount of withaferin A or a salt thereof, a therapeutically effective amount of tilorixan or a salt thereof, and a pharmaceutically acceptable carrier or excipient.

3. The pharmaceutical composition according to claim 1 or 2, comprising about 1 mg to 4000 mg of withaferin A or a salt thereof.

4. The pharmaceutical composition according to any one of claims 1 to 3, comprising about 1 mg to 4000 mg of tilorixen or a salt thereof.

5. The pharmaceutical composition according to any one of claims 1 to 4, wherein the dosage ratio of withaferin A or its salt and tilorixan or its salt is about 10:1 to about 1:10, preferably about 1:1 or about 1:

2.

6. The pharmaceutical composition according to any one of claims 1 to 5, wherein the pharmaceutical composition is used to prevent or treat a neurodegenerative disease in a subject.

7. The pharmaceutical composition according to claim 6, wherein the neurodegenerative disease is selected from Alzheimer's disease, Parkinson's disease, or Huntington's disease.

8. Use of a pharmaceutical composition for preparing a medicament for preventing or treating a neurodegenerative disease in a subject, wherein the pharmaceutical composition comprises i) withaferin A or a salt thereof; and ii) tilorixan or a salt thereof.

9. The use according to claim 8, wherein the pharmaceutical composition comprises a therapeutically effective amount of withaferin A or a salt thereof, a therapeutically effective amount of tilorixan or a salt thereof, and a pharmaceutically acceptable carrier or excipient.

10. The use according to claim 8 or 9, wherein the pharmaceutical composition comprises about 1 mg to 4000 mg of withaferin A or a salt thereof.

11. The use according to any one of claims 8 to 10, wherein the pharmaceutical composition comprises about 1 mg to 4000 mg of tilorixen or a salt thereof.

12. The use according to any one of claims 8 to 11, wherein in the pharmaceutical composition, the dosage ratio of withaferin A or its salt to tilorixan or its salt is about 10:1 to about 1:10, preferably about 1:1 or about 1:

2.

13. The use according to any one of claims 8 to 12, wherein the neurodegenerative disease is selected from Alzheimer's disease, Parkinson's disease, or Huntington's disease.

14. Use of withaferin A or a salt thereof in preparing a combined pharmaceutical composition for preventing or treating a neurodegenerative disease in a subject, wherein the combined pharmaceutical composition comprises i) withaferin A or a salt thereof; and ii) tilorixan or a salt thereof.

15. Use of tilorixan or a salt thereof in preparing a combination pharmaceutical composition for preventing or treating a neurodegenerative disease in a subject, wherein the combination pharmaceutical composition comprises i) withaferin A or a salt thereof; and ii) tilorixan or a salt thereof.

16. The use according to claim 14 or 15, wherein the combined pharmaceutical composition comprises a therapeutically effective amount of withaferin A or a salt thereof, a therapeutically effective amount of tilorixan or a salt thereof, and a pharmaceutically acceptable carrier or excipient.

17. The use according to any one of claims 14 to 16, wherein the pharmaceutical composition comprises about 1 mg to 4000 mg of withaferin A or a salt thereof.

18. The use according to any one of claims 14 to 17, wherein the pharmaceutical composition comprises about 1 mg to 4000 mg of tilorixen or a salt thereof.

19. The use according to any one of claims 14 to 18, wherein in the pharmaceutical composition, the dosage ratio of withaferin A or its salt and tilorixan or its salt is about 10:1 to about 1:10, preferably about 1:1 or about 1:

2.

20. The use according to any one of claims 14 to 19, wherein the neurodegenerative disease is selected from Alzheimer's disease, Parkinson's disease, or Huntington's disease.

21. A method for preventing or treating a neurodegenerative disease in a subject in need thereof, comprising administering to the subject in need thereof a therapeutically effective amount of withaferin A or a salt thereof, and administering to the subject a therapeutically effective amount of tilorixan or a salt thereof.

22. The method according to claim 21, wherein the administration of withaferin A or a salt thereof and tilorixan or a salt thereof is performed simultaneously or sequentially in any order.

23. The method according to claim 21 or 22, wherein withaferin A or a salt thereof is administered in an amount of about 0.02-40 mg / kg, preferably about 2-4 mg / kg, daily or once every two days.

24. The method according to any one of claims 21 to 23, wherein tilorixen or a salt thereof is administered in an amount of about 0.02-40 mg / kg, preferably about 2-10 mg / kg, daily or once every two days.

25. The method according to any one of claims 21 to 24, wherein the administered doses of withaferin A or its salt and tilorixan or its salt are in a ratio of about 10:1 to about 1:10, preferably about 1:1 or 1:

2.

26. The method of any one of claims 21 to 25, wherein the neurodegenerative disease is selected from Alzheimer's disease, Parkinson's disease, or Huntington's disease.

27. Use of withaferin A or a salt thereof for enhancing the effect of tilorixen or a salt thereof in preventing or treating a neurodegenerative disease in a subject.

28. Use of tilorixan or a salt thereof for enhancing the effect of withaferin A or a salt thereof in preventing or treating a neurodegenerative disease in a subject.

29. Use of withaferin A or a salt thereof in the preparation of a synergist for tilorixen or a salt thereof in preventing or treating neurodegenerative diseases.

30. Use of tilorixan or a salt thereof in the preparation of a synergist for withaferin A or a salt thereof in preventing or treating neurodegenerative diseases.

31. The use according to claim 27 or 29, wherein the amount of withaferin A or its salt is about 1 mg to 4000 mg.

32. The use according to claim 28 or 30, wherein the amount of tilorixen or a salt thereof is about 1 mg to 4000 mg.

33. The use according to any one of claims 27 to 30, wherein the dosage ratio of withaferin A or its salt and tilorixan or its salt is from about 10:1 to about 1:10, preferably about 1:1 or about 1:

2.

34. Use according to any one of claims 27 to 33, wherein the neurodegenerative disease is selected from Alzheimer's disease, Parkinson's disease, or Huntington's disease.

35. The pharmaceutical composition according to claim 6 or 7, the use according to any one of claims 8 to 19 or any one of claims 27 to 33, the method according to any one of claims 21 to 26, wherein the subject is a mammal, preferably a human subject.

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