WO2023036105A1

WO2023036105A1 - Method for treating neurodegenerative disease

Info

Publication number: WO2023036105A1
Application number: PCT/CN2022/117178
Authority: WO
Inventors: 张寰; 桑绍明
Original assignee: 上海日馨医药科技股份有限公司
Priority date: 2021-09-09
Filing date: 2022-09-06
Publication date: 2023-03-16

Abstract

An application of a pharmaceutical composition composed of a first medicament and a second medicament in the preparation of a drug for preventing or treating a neurodegenerative disease, alleviating symptoms of the neurodegenerative disease, or slowing down the progression of the neurodegenerative disease. The first medicament is benfotiamine or a pharmaceutically acceptable salt thereof, and the second medicament is an acetylcholinesterase inhibitor that is donepezil, rivastigmine, galantamine, tacrine, or a pharmaceutically acceptable salt thereof.

Description

Methods of treating neurodegenerative diseases

field of invention

The present invention belongs to the field of biomedicine, and in particular relates to methods for preventing or treating neurodegenerative diseases, alleviating symptoms of neurodegenerative diseases, or delaying the progress of neurodegenerative diseases, which comprises administering a preventive or therapeutically effective dose to individuals in need thereof A first agent and a prophylactically or therapeutically effective amount of a second agent, wherein the first agent is benfotiamine or a pharmaceutically acceptable salt thereof; and the second agent is an acetylcholinesterase inhibitor.

Background of the invention

Alzheimer's disease (AD) is the most common degenerative disease of the central nervous system. Due to the large number of patients, the protracted course of the disease, the long survival period of patients who lose their ability to take care of themselves, and the lack of effective prevention and treatment drugs, it has brought huge economic and spiritual burdens to individuals, families and even the whole society. According to reports, in 2015, China spent up to 167.74 billion US dollars on AD prevention and care, and it is expected that the cost will reach 507.49 billion US dollars by 2030. In 2018, the global cost of AD-based dementia diseases exceeded one trillion U.S. dollars, accounting for more than 1% of the global gross domestic product (GDP). AD is the only disease that lacks effective prevention and treatment drugs among the top ten major diseases in the world. It has become a major disease that seriously affects the health care system and sustainable economic development of major economies in the world, including China.

AD is a disease with multiple pathophysiological changes, including neuronal loss, glial cell activation, and characteristic senile plaques formed by extracellular β-amyloid (Aβ) deposition, and neurofibrillary tangles caused by hyperphosphorylation of intracellular Tau protein. Knot and so on. In addition, synaptic loss, brain glucose metabolism disorder, and oxidative stress are also constant pathological changes in the AD brain, and the decline in brain glucose metabolism is closely related to cognitive dysfunction. Due to the unclear pathogenesis, AD still lacks effective treatments.

Summary of the invention

In one aspect, the application provides methods for preventing or treating neurodegenerative diseases, reducing symptoms of neurodegenerative diseases, or delaying the progress of neurodegenerative diseases, which includes administering a preventive or therapeutically effective amount of the first An agent and a prophylactically or therapeutically effective amount of a second agent, wherein the first agent is benfotiamine or a pharmaceutically acceptable salt thereof; and the second agent is an acetylcholinesterase inhibitor.

In another aspect, the present application provides the preparation of the first medicament for preventing or treating neurodegenerative diseases, alleviating the symptoms of neurodegenerative diseases, or delaying the progress of neurodegenerative diseases in combination with the second medicament. Uses in medicine.

In another aspect, the present application provides the first medicament for use in combination with the second medicament to prevent or treat neurodegenerative diseases, alleviate symptoms of neurodegenerative diseases, or delay the progression of neurodegenerative diseases.

The neurodegenerative disease is preferably Alzheimer's disease; more preferably, the Alzheimer's disease is mild to moderate Alzheimer's disease.

Brief description of the drawings

Fig. 1 is the impact of each compound on the incubation period of animals in the embodiment;

Fig. 2 is the impact of each compound on the number of times of animal crossing in the embodiment;

Fig. 3 is the effect of each compound in the Examples on the target quadrant time of animals.

Detailed description of the invention

definition

Unless defined otherwise hereinafter, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. References to techniques used herein are intended to refer to techniques commonly understood in the art, including those variations of the techniques or substitutions of equivalent techniques that are obvious to those skilled in the art. While the following terms are believed to be well understood by those skilled in the art, the following definitions are set forth to better explain the present invention.

The terms "comprising", "comprising", "having", "containing" or "involving" and other variations thereof herein are inclusive or open-ended and do not exclude other unlisted elements or method steps.

Donepezil, molecular formula C ₂₄ H ₂₉ NO ₃ , structural formula is

Its common medicinal form is donepezil hydrochloride, which is the main drug for the clinical treatment of AD. It increases the level of acetylcholine by inhibiting cholinesterase to delay the progression of AD in the middle and early stages and improve the clinical symptoms of early AD.

Benfotiamine (benzoylthiamineomonophosphate, referred to as BTMP), the molecular formula C ₁₉ H ₂₃ N ₄ O ₆ PS, the structural formula is

Benfotiamine can improve the shortcoming of low bioavailability of water-soluble vitamin B1, increase the concentration of vitamin B1 in blood and tissues, and thus improve the curative effect.

The term "pharmaceutically acceptable salt" includes acid addition salts and base addition salts thereof.

Suitable acid addition salts are formed from acids which form pharmaceutically acceptable salts. Examples include acetate, adipate, aspartate, benzoate, benzenesulfonate, bicarbonate/carbonate, bisulfate/sulfate, borate, camphorsulfonate , citrate, cyclamate, edisylate, ethanesulfonate, formate, fumarate, glucoheptonate, gluconate, glucuronate, hexafluorophosphate Salt, seabenzoate, hydrochloride/chloride, hydrobromide/bromide, hydroiodide/iodide, isethionate, lactate, malate, maleic acid Salt, malonate, methanesulfonate, methylsulfate, naphthylate, 2-naphthalenesulfonate, nicotinate, nitrate, orotate, oxalate, palmitic acid Salt, Pamoate, Phosphate/Hydrogen Phosphate/Dihydrogen Phosphate, Pyroglutamate, Saccharate, Stearate, Succinate, Tannin, Tartrate, Toluene Sulfonate, trifluoroacetate and xinafoate.

Suitable base addition salts are formed from bases which form pharmaceutically acceptable salts. Examples include aluminum salts, arginine salts, benzathine penicillin salts, calcium salts, choline salts, diethylamine salts, diethanolamine salts, glycinate salts, lysine salts, magnesium salts, meglumine salts, ethanolamine salts, Potassium, sodium, tromethamine and zinc salts.

For a review of suitable salts see "Handbook of Pharmaceutical Salts: Properties, Selection, and Use" by Stahl and Wermuth (Wiley-VCH, 2002). Methods for preparing pharmaceutically acceptable salts of the compounds of the invention are known to those skilled in the art.

The term "about" means within ±10%, preferably within ±5%, more preferably within ±2% of the stated value.

In the context of the present invention, when said first agent and said second agent are used in combination, this means that in one embodiment, said two compounds may be administered simultaneously. In another embodiment, when said first agent and said second agent are used in combination, this means that said two compounds are administered separately in suitable separate pharmaceutical compositions. The separate compositions can be administered simultaneously, e.g., in the morning or evening, once a day, at regular intervals; or they can be administered independently, e.g., one compound twice a day after breakfast and dinner , at regular intervals, and the other compound is administered at regular intervals once a day after dinner.

The term "prevention" as used herein refers to the prior administration of a drug to avoid or prevent the occurrence of one or more symptoms of a disease or disorder. Those of ordinary skill in the medical arts recognize that the term "prophylaxis" is not an absolute term. In the medical field, prophylactic administration of a drug is understood to mean substantially reducing the likelihood or severity of a disorder or symptoms of a disorder, and this is the intended meaning in this disclosure. Prevention is divided into primary prevention (to prevent the development of the disease) and secondary prevention (whereby the disease has already developed and the patient is protected from worsening of the process).

As used herein, unless otherwise stated, the term "treating" means reversing, alleviating, inhibiting the disorder or condition to which such term applies or the progression of one or more symptoms of such disorder or condition, or Such a disorder or condition or one or more symptoms of such a disorder or condition is prevented.

For example, the term "treating" may include reducing or alleviating cognitive impairment (such as impaired memory and/or orientation) or overall functional impairment (activities of daily living (ADL)) and/or slowing or reversing progressive deterioration in ADL or Cognitive impairment in individuals with mild to moderate Alzheimer's disease. The term "treating" can also refer to delaying disease progression in patients with additional symptoms associated with Alzheimer's disease, such as, but not limited to, using ADAS-cog, MMSE, ADCS-ADL criteria, CDR-sb, or NPI overall criteria One or more of those identified.

As used herein, the term "delayed progression" refers to a slower than expected progression or persistence or worsening of an individual's disease compared to untreated patients or patients treated with only a single agent (eg, donepezil). can be determined using, for example, the Alzheimer's Disease Assessment Scale (ADAS-cog) or the Alzheimer's Disease Cooperative Study-Activities of Daily Living (ADCS-ADL) cognitive performance.

"Individual" as used herein includes a human or non-human animal. Exemplary human subjects include human subjects suffering from a disease (eg, a disease described herein) (referred to as a patient) or normal subjects. "Non-human animals" in the present invention include all vertebrates, such as non-mammals (e.g., birds, amphibians, reptiles) and mammals, such as non-human primates, livestock and/or domesticated animals (e.g., sheep, dogs, , cats, cows, pigs, etc.).

An "effective amount" is a clinically observable amount associated with Alzheimer's disease (as measured by ADAS-cog) (Rosen WG et al., A new scale for Alzheimer's disease, Am J Psychiatry, 1984; 141:1356-64). An amount sufficient to provide an observable prophylactic or therapeutic benefit compared to a baseline of observed signs and symptoms.

ADAS-cog was measured according to Huali Wang et al. The cognitive subscale of Alzheimer's Disease Assessment Scale, Chinese version in staging of Alzheimer disease, Alzheimer Dis Assoc Disord. Oct-Dec 2004; 18(4): 231-5.

ADCS-ADL was measured according to Galasko et al. An inventory to assess activities of daily living for clinical trials in Alzheimer's disease. The Alzheimer's Disease Cooperative Study. Alzheimer Dis Assoc Disord. 1997; 11 Suppl 2: S33-9.

According to Cummings, J.L., Mega, M., Gray, K., Rosenberg-Thompson, S., Carusi, D.A., and Gornbein, J. The Neuropsychiatric Inventory: comprehensive assessment of psychopathology in dementia. Neurology, 1994;44:2308-2314 , measuring the Neuropsychiatric Inventory (NPI).

According to Folstein, M.F., Folstein, S.E. and McHugh, P.R. "Mini-mental state": a practical method for grading the cognitive state of patients for the clinician. Journal of psychiatric research, 1975 12(3):189-198, easy to measure Mental State Examination (MMSE).

The term "mild Alzheimer's disease" may mean a score > 20 on the Mini-Mental State Examination (MMSE) scale. The term "moderate Alzheimer's disease" may mean a score on the Mini-Mental State Examination (MMSE) scale between 11 and 19, inclusive.

By "reduced rate of deterioration in score" is meant a decrease in the change in score from baseline in the treatment group compared to the change in score from baseline in the placebo group (or without treatment).

In some embodiments, the present application provides methods for preventing or treating neurodegenerative diseases, reducing symptoms of neurodegenerative diseases, or delaying the progress of neurodegenerative diseases, comprising administering a preventive or therapeutically effective amount of A first agent and a prophylactically or therapeutically effective amount of a second agent, wherein the first agent is benfotiamine or a pharmaceutically acceptable salt thereof; and the second agent is an acetylcholinesterase inhibitor.

In some embodiments, the first agent and the second agent are administered separately.

In another embodiment, the present invention provides the use of a first agent in the preparation of a medicament for preventing or treating a neurodegenerative disease, alleviating the symptoms of a neurodegenerative disease, or delaying the progression of a neurodegenerative disease in combination with a second agent. use.

In another embodiment, the present invention provides the use of a second agent in the preparation of a medicament for preventing or treating a neurodegenerative disease, alleviating the symptoms of a neurodegenerative disease, or delaying the progression of a neurodegenerative disease in combination with the first agent. use.

In another embodiment, the present invention provides the use of a first agent in the manufacture of a neurodegenerative disease in a subject being treated with a therapy comprising a second agent, to alleviate the symptoms of said neurodegenerative disease, or to delay the Use in medicines for the progression of neurodegenerative diseases.

In another embodiment, the present invention provides a second agent in the manufacture of a neurodegenerative disease for the prevention or treatment of a subject being treated with a therapy comprising the first agent, to alleviate the symptoms of said neurodegenerative disease, or to delay the Use in medicines for the progression of neurodegenerative diseases.

In another embodiment, the present invention provides a first agent for use in combination with a second agent to prevent or treat a neurodegenerative disease, alleviate the symptoms of a neurodegenerative disease, or delay the progression of a neurodegenerative disease.

In another embodiment, the present invention provides a second agent for use in combination with a first agent to prevent or treat a neurodegenerative disease, alleviate the symptoms of a neurodegenerative disease, or delay the progression of a neurodegenerative disease.

In another embodiment, the present invention provides a first agent for preventing or treating a neurodegenerative disease, alleviating the symptoms of said neurodegenerative disease, or delaying progression of neurodegenerative diseases.

In another embodiment, the present invention provides a second agent for preventing or treating a neurodegenerative disease, alleviating the symptoms of said neurodegenerative disease, or delaying progression of neurodegenerative diseases.

In some embodiments, the neurodegenerative disease is Alzheimer's disease, more preferably, the Alzheimer's disease is mild to moderate Alzheimer's disease.

In various embodiments, the first agent is benfotiamine, or a pharmaceutically acceptable salt thereof; and the second agent is an acetylcholinesterase inhibitor.

In some embodiments, the second agent is donepezil, rivastigmine, galantamine, tacrine or a pharmaceutically acceptable salt thereof; preferably donepezil or a pharmaceutically acceptable salt thereof; most Preferred is donepezil hydrochloride.

In some embodiments, the first agent is administered in an amount of about 0.005 mg/day to about 5000 mg/day, such as about 0.005, 0.05, 0.5, 5, 10, 20, 30, 40, 50, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, 1000, 1500, 2000, 2500, 3000, 3500, 4000, 4500 or 5000mg/ daily dosage.

In a preferred embodiment, the first agent is administered in an amount of about 600 mg/day.

In some embodiments, the first agent is administered in an amount of about 1 ng/kg to about 200 mg/kg, about 1 μg/kg to about 100 mg/kg, or about 1 mg/kg to about 50 mg/kg body weight per day, For example, at about 1 μg/kg, about 10 μg/kg, about 25 μg/kg, about 50 μg/kg, about 75 μg/kg, about 100 μg/kg, about 125 μg/kg, about 150 μg/kg, about 175 μg/kg, about 200 μg/kg, about 225 μg/kg, about 250 μg/kg, about 275 μg/kg, about 300 μg/kg, about 325 μg/kg, about 350 μg/kg, about 375 μg/kg, about 400 μg/kg, about 425 μg/kg, about 450 μg/kg, about 475 μg/kg, about 500 μg/kg, about 525 μg/kg, about 550 μg/kg, about 575 μg/kg, about 600 μg/kg, about 625 μg/kg, about 650 μg/kg, about 675 μg/kg, about 700 μg/kg, about 725 μg/kg, about 750 μg/kg, about 775 μg/kg, about 800 μg/kg, about 825 μg/kg, about 850 μg/kg, about 875 μg/kg, about 900 μg/kg, about 925 μg/kg, about 950 μg/kg, about 975 μg/kg, about 1 mg/kg, about 5 mg/kg, about 10 mg/kg, about 15 mg/kg, about 20 mg/kg, about 25 mg/kg, about 30 mg/kg, about 35 mg/kg, about 40mg/kg, about 45mg/kg, about 50mg/kg, about 60mg/kg, about 70mg/kg, about 80mg/kg, about 90mg/kg, about 100mg/kg, about 125mg/kg, about 150mg/kg, about An amount of 175 mg/kg, about 200 mg/kg or about 300 mg/kg of body weight is administered.

In some embodiments, the second agent is administered in an amount of about 0.005 mg/day to about 5000 mg/day, such as about 0.005, 0.05, 0.5, 5, 10, 20, 30, 40, 50, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, 1000, 1500, 2000, 2500, 3000, 3500, 4000, 4500 or 5000mg/ daily dosage.

In a preferred embodiment, the second agent is administered in an amount of about 5 mg/day.

In some embodiments, the second agent is administered in an amount of about 1 ng/kg to about 200 mg/kg, about 1 μg/kg to about 100 mg/kg, or about 1 mg/kg to about 50 mg/kg body weight per day, For example, at about 1 μg/kg, about 10 μg/kg, about 25 μg/kg, about 50 μg/kg, about 75 μg/kg, about 100 μg/kg, about 125 μg/kg, about 150 μg/kg, about 175 μg/kg, about 200 μg/kg, about 225 μg/kg, about 250 μg/kg, about 275 μg/kg, about 300 μg/kg, about 325 μg/kg, about 350 μg/kg, about 375 μg/kg, about 400 μg/kg, about 425 μg/kg, about 450 μg/kg, about 475 μg/kg, about 500 μg/kg, about 525 μg/kg, about 550 μg/kg, about 575 μg/kg, about 600 μg/kg, about 625 μg/kg, about 650 μg/kg, about 675 μg/kg, about 700 μg/kg, about 725 μg/kg, about 750 μg/kg, about 775 μg/kg, about 800 μg/kg, about 825 μg/kg, about 850 μg/kg, about 875 μg/kg, about 900 μg/kg, about 925 μg/kg, about 950 μg/kg, about 975 μg/kg, about 1 mg/kg, about 5 mg/kg, about 10 mg/kg, about 15 mg/kg, about 20 mg/kg, about 25 mg/kg, about 30 mg/kg, about 35 mg/kg, about 40mg/kg, about 45mg/kg, about 50mg/kg, about 60mg/kg, about 70mg/kg, about 80mg/kg, about 90mg/kg, about 100mg/kg, about 125mg/kg, about 150mg/kg, about An amount of 175 mg/kg, about 200 mg/kg or about 300 mg/kg of body weight is administered.

In some embodiments, the daily dose of the first agent and/or the second agent is administered in one dose or in two, three or four divided doses.

In a preferred embodiment, the daily dose of the first agent is administered in two divided doses.

In a preferred embodiment, the daily dose of said second agent is administered as a single dose.

In some embodiments, the first agent and/or the second agent are administered continuously for at least 1 week, at least 2 weeks, at least 3 weeks, at least 4 weeks, at least 5 weeks, at least 6 weeks, at least 7 weeks , at least 8 weeks, at least 9 weeks, at least 10 weeks, at least 11 weeks, at least 12 weeks, at least 13 weeks, at least 14 weeks, at least 15 weeks, at least 16 weeks, at least 17 weeks, at least 18 weeks, at least 19 weeks, at least 20 weeks, at least 21 weeks, at least 22 weeks, at least 23 weeks, at least 24 weeks, at least 25 weeks, at least 30 weeks, at least 35 weeks, at least 40 weeks, at least 45 weeks, at least 50 weeks, at least 51 weeks, at least 52 weeks , at least 53 weeks, at least 54 weeks, or at least 55 weeks.

In some embodiments, the first agent and/or the second agent are administered for one or more (eg, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10) courses of treatment , wherein each course of treatment lasts at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 8 days, at least 9 days, at least 10 days, at least 11 days, at least 12 days, at least 13 days, At least 14 days, at least 15 days, at least 16 days, at least 17 days, at least 18 days, at least 19 days, at least 20 days, at least 21 days, at least 22 days, at least 23 days, at least 24 days, at least 25 days, at least 30 days days, at least 35 days, at least 40 days, at least 45 days, or at least 50 days; week, three weeks or four weeks.

In some embodiments, said first agent and/or said second agent is administered by injection (e.g., intravenous, intraarterial, subcutaneous, intraperitoneal, intramuscular injection, including instillation) or transdermally; or Administration is oral, buccal, nasal, transmucosal, topically, in the form of ophthalmic preparations or by inhalation.

In a preferred embodiment, said first agent and said second agent are administered orally.

In some embodiments, the first medicament and/or the second medicament are selected from tablets, capsules, lozenges, hard lozenges, powders, sprays, creams, ointments, suppositories, gelatin Dosage forms of gels, pastes, lotions, ointments, aqueous suspensions, injectable solutions, elixirs, and syrups are administered.

In a preferred embodiment, said first agent and said second agent are administered in the dosage form of a tablet.

In some embodiments, the method improves the following pathophysiological manifestations in the individual: cognitive behavioral abnormalities, neurodegenerative changes (e.g., progressive synapse/neuron loss and brain atrophy), β-amyloid deposition, Tau Aberrant phosphorylation and its resulting neurofibrillary tangles, glial cell activation and inflammation and/or impaired glucose metabolism in the brain.

In some embodiments, the second agent is administered in a lower amount than the amount administered when the second agent is administered alone or in the absence of the first agent.

In some embodiments, the first agent enhances the efficacy of the second agent in treating a neurodegenerative disease and/or reduces the side effects of the second agent in treating a neurodegenerative disease.

In some embodiments, said individual has received monotherapy with said second agent prior to receiving the combined administration of said first agent and said second agent.

In some embodiments, the method results in an improvement or no deterioration or a reduced rate of deterioration in at least one assessment selected from: Alzheimer's Disease Assessment Scale-Cognitive Subscale (ADAS-cog); Clinical Dementia Rating Pencil Box (CDR-sb); Alzheimer's Disease Collaborative Study Activities of Daily Living Scale (ADCS-ADL); Neuropsychiatric Inventory (NPI); and Mini-Mental State Examination (MMSE).

In some embodiments, the method results in a decreased rate of deterioration in the ADAS-cog score.

In some embodiments, the method reduces the rate of deterioration of the ADAS-cog score by an average of 1-5 points, preferably 2-3.5 points.

The present invention encompasses any combination of the above embodiments.

Example

Embodiment 1.Morris water maze experiment

1. Experimental principle

Rodents have a strong motivation to escape from the water environment in water, and they can escape from the water environment in the fastest and most direct way. The process of learning to escape from the water environment reflects the learning ability of animals. Space positioning according to the surrounding environment and purposeful swimming to a safe place in the water (such as a platform) can reflect the spatial learning and memory ability of animals.

2. Experimental content

2.1. Materials

(1) Experimental animals

The experimental animals were male APP/PS1/Tau mice and C57BL/6 wild-type mice. Rats aged 8 months and weighing 250-300 g were purchased from The Jackson laboratory.

APP/PS1/Tau mice are APP/PS1/tau triple transgenic Alzheimer's disease (3×Tg-AD) model mice. The appearance of Aβ deposition is several months earlier than the pathological changes of tau protein, which can more realistically simulate Clinical course and pathological changes of Alzheimer's disease.

(2) Main preparations

CMC-Na (sodium carboxymethylcellulose), Shanghai Sinopharm Chemical Reagent Co., Ltd., batch number 20181203; BTMP (benfotiamine), Shanghai Rixin Biotechnology Co., Ltd., batch number 1908002; donepezil hydrochloride, Eisai (China) Manufactured by Pharmaceutical Co., Ltd., lot number 1704019.

2.2. Drug preparation and administration information

(1) Drug preparation

Preparation of 0.7% m/v CMC-Na (sodium carboxymethyl cellulose): Weigh 0.7g sodium carboxymethyl cellulose and dissolve it in 100ml of purified water to obtain 0.7% m/v CMC-Na.

Preparation of 10mg/ml vitamin B1 solution (dosage 200mg/kg): Weigh 500mg vitamin B1, add 0.7% m/v CMC-Na to 50ml, and obtain 10mg/ml solution.

Preparation of 10mg/ml BTMP suspension (dosage 200mg/kg): Weigh 500mg BTMP, add 0.7% m/v CMC-Na to 50ml, and obtain 10mg/ml BTMP suspension.

Preparation of 20mg/ml BTMP suspension (dosage 400mg/kg): Weigh 500mg BTMP, add 0.7% m/v CMC-Na to 25ml, and obtain 20mg/ml BTMP suspension.

Preparation of 0.075mg/ml donepezil hydrochloride suspension (dosage 1.5mg/kg): 1 piece of donepezil hydrochloride tablet (containing 5mg donepezil hydrochloride) was ground and added 0.7% m/v CMC-Na to 66.67ml to obtain 0.075 mg/ml donepezil hydrochloride suspension.

Preparation of 0.15mg/ml donepezil hydrochloride suspension (administration dose: 3.0mg/kg): 1 piece of donepezil hydrochloride tablet (containing 5mg donepezil hydrochloride) was ground and added with 0.7% m/v CMC-Na to 33.33ml to obtain 0.15 mg/ml donepezil hydrochloride suspension.

Preparation of 10mg/ml BTMP suspension + 0.075mg/ml donepezil hydrochloride suspension (dose 200mg/kg BTMP suspension + 1.5mg/kg donepezil hydrochloride suspension): prepare 20mg/ml BTMP suspension respectively liquid and 0.15mg/ml donepezil hydrochloride suspension, mixed in equal volumes before administration to obtain 10mg/ml BTMP suspension + 0.075mg/ml donepezil hydrochloride suspension.

Preparation of 5mg/ml BTMP suspension + 0.075mg/ml donepezil hydrochloride suspension (dose 100mg/kg BTMP suspension + 1.5mg/kg donepezil hydrochloride suspension): prepare 10mg/ml BTMP suspension respectively liquid and 0.15mg/ml donepezil hydrochloride suspension, mixed in equal volumes before administration to obtain 5mg/ml BTMP suspension + 0.075mg/ml donepezil hydrochloride suspension.

(2) Administration information

2.3. Experimental method

The mice in the blank control group, the model group and the experimental group were administered orally for 8 weeks according to 2.2. Dosage/specification, and water maze training and testing began in the last week of administration. The water maze training and testing lasted for 6 days, the training period was 5 days, and the testing period was 1 day. In addition, during the water maze training and testing period (a total of 6 days), keep the indoor lighting and other conditions consistent, keep the room quiet, and eliminate the environment, personnel and other interference.

(1) Preparation before the experiment: Put an appropriate amount of water in the water maze pool, keep the water temperature at 22±3°C, place the platform at a fixed position (target quadrant) and 1em below the water surface, add titanium dioxide until the water color is mixed with white, and the platform cannot be seen clearly .

(2) Training period (day 1 to day 5): Before the first quadrant training every day, each mouse was placed on the platform for 15s (increasing the mouse's sense of security on the platform), and then the mice were removed from the platform. Put the quadrant into the pool (head facing the pool wall), and set the swimming time to 60s. If it finds the platform within 60s and stays for 5s, it is deemed to have successfully found the platform. If the platform cannot be found, the recording time is 60s, the mouse is guided to the platform and stays for 20s, the mouse is taken out, and the quadrant training of this mouse is ended.

After the training in the first quadrant, the training in the remaining three quadrants was performed sequentially, and it was not necessary to place the mice on the platform for 15 seconds before the training. The training time interval between two quadrants of each mouse was 10-15 minutes. In this way of training, continuous training for 5 days.

(3) Test period (Day 6): 24 hours after the last training session, the platform was removed, and the mice were dropped on the opposite side of the quadrant of the original platform (that is, the farthest position from the platform). The quadrant where the platform is located) time, the times of passing through the original platform position (the number of times of crossing the platform) and the time of passing through the original platform position for the first time (latency period) are used as the indicators for the investigation of the mouse's spatial learning and memory ability.

3. Experimental results

Compared with the blank control group, the latency time of the mice in the model group was significantly increased, the times of crossing the platform were less, and the time in the target quadrant was significantly reduced, and there was a significant difference. It shows that there are differences between the transgenic model mice and normal mice in the ability of spatial learning and memory, and is suitable for training in spatial learning and memory. Compared with the model group, the incubation period, times of platform crossing and target quadrant time of the positive control group and the experimental group were significantly improved, and there were significant differences. The results are shown in Table 1.

Table 1 Morris water maze test results for mice in each compound group (n=10)

Remarks: P<0.05 means significant difference; *P<0.05-compared with blank control group; ^# P<0.05-compared with model group; ^& P<0.05-compared with BTMP

Taking the incubation period, times of crossing the platform, and target quadrant time as the indicators for the drug efficacy of the compound, the order is blank control group, BTMP+ donepezil hydrochloride, 1/2BTMP+ donepezil hydrochloride, BTMP, donepezil hydrochloride, VB1, model group (the one with the best performance) . The results are shown in Figures 1 to 3. The results show that VB1 does not significantly improve the learning and memory of AD mice, and BTMP, donepezil hydrochloride, BTMP+ donepezil hydrochloride, and 1/2BTMP+ donepezil hydrochloride have significant improvements in the spatial memory learning ability of AD mice. And the combined administration of BTMP and donepezil hydrochloride has the best improvement effect, which is obviously better than that of BTMP and donepezil hydrochloride alone.

The results of the water maze experiment of the present invention show that after the combined administration of benfotiamine and donepezil hydrochloride, a better synergistic effect can be achieved, which can better improve the spatial learning and memory ability of AD mice, and the effect is significantly better than that of benfotiamine Or the effect of donepezil hydrochloride when used alone is also better than the effect of vitamin B1, and the pharmaceutical composition of the present invention has better drug activity in the treatment of Alzheimer's disease.

Test Example 2: Clinical Trial

A 52-week clinical trial study was conducted in patients with moderate Alzheimer's disease (MMSE 11-19 at screening). The study groups and dosing regimen are as follows:

Based on the ADAS-cog scale (including 11 items in total (word recall, naming, executive instructions, structural exercises, intentional exercises, orientation, word recognition, recall test instructions, oral language skills, word finding skills, language comprehension skills) ), used to evaluate the level of cognitive function of patients, the score ranges from 0 to 70 points, the higher the score indicates the more severe cognitive impairment), the patients were scored after 12, 24, 36, and 52 weeks of administration. The between-group differences in patient score changes from baseline are shown in the table below.

From the above results, it can be seen that the improvement of ADAS-cog score of patients in the treatment group was better than that in the placebo group. In particular, the ADAS-cog score of the patients in the treatment group improved by 2.116 points at 36 weeks, with a statistical difference (p=0.077), and the ADAS-cog score of the patients in the 52-week treatment group improved by 3.331 points, with a statistical difference (p=0.029). The results showed that the combination therapy of benfotiamine and donepezil hydrochloride can significantly delay the progression of Alzheimer's disease.

At the same time, during the administration period, no obvious serious drug-related adverse reactions were observed in the patients in the treatment group, indicating that the combination therapy of benfotiamine and donepezil hydrochloride has good safety, and the patients tolerated the therapy well.

Test Example 3: Clinical Trial

A 52-week clinical trial study was conducted in patients with moderate Alzheimer's disease (patients with an MMSE of 11-19 at the time of enrollment screening were classified as moderate AD patients). The study grouping, dosing regimen and the number of patients who completed the experiment in each group are as follows:

Apolipoprotein E (APOE) genotype was measured before administration, and AD patients were divided into APOE ε4 allele carrier group and non-APOE ε4 allele carrier group.

Based on the ADAS-cog scale as described in Test Example 2, the ADAS-cog score was performed on the patient after 52 weeks of administration, and the changes in the patient's score relative to the baseline are shown in the table below.

*: p<0.05, compared with placebo group

It can be seen from the above table that compared with the placebo group, the average change in the ADAS-cog score of non-APOE ε4 allele carriers in the high-dose group was -5.09 (p<0.05), while the ADAS score of APOE ε4 allele carriers The mean change in -cog score was -1.09.

From the above results, it can be seen that whether the treated patients carry the APOE ε4 allele has a significant impact on the effect of benfotiamine/donepezil combined treatment of AD. The therapeutic effect on non-APOE ε4 allele carriers was significantly better than that of APOE ε4 allele carriers.

Various modifications of the invention, in addition to those described herein, will be apparent to those skilled in the art from the foregoing description. Such modifications are also intended to fall within the scope of the appended claims. Each reference cited in this application, including all patents, patent applications, journal articles, books, and any other publications, is hereby incorporated by reference in its entirety.

Claims

A method for preventing or treating a neurodegenerative disease, alleviating the symptoms of a neurodegenerative disease, or delaying the progression of a neurodegenerative disease, comprising administering a prophylactically or therapeutically effective amount of a first agent and a prophylactically or therapeutically effective amount of a first agent to an individual in need thereof the second dose of

wherein the first agent is benfotiamine or a pharmaceutically acceptable salt thereof; and the second agent is an acetylcholinesterase inhibitor;

Preferably, said first agent and said second agent are administered separately;

Preferably, the neurodegenerative disease is Alzheimer's disease, more preferably, the Alzheimer's disease is mild to moderate Alzheimer's disease.
The method of claim 1, wherein the second agent is donepezil, rivastigmine, galantamine, tacrine or a pharmaceutically acceptable salt thereof; preferably donepezil or a pharmaceutically acceptable salt thereof; Most preferred is donepezil hydrochloride.
The method of claim 1 or 2, wherein the first agent is administered in an amount of about 0.005 mg/day to about 5000 mg/day, such as about 0.005, 0.05, 0.5, 5, 10, 20, 30, 40, 50, 100 , 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, 1000, 1500, 2000, 2500, 3000, 3500, 4000, 4500 Or 5000mg/day dose administration;

Preferably, the first agent is administered in an amount of about 600 mg/day.
The method of any one of claims 1-3, wherein the first agent is administered at about 1 ng/kg to about 200 mg/kg, about 1 μg/kg to about 100 mg/kg, or about 1 mg/kg to about 50 mg/kg per day. Administration in an amount of kg body weight, for example, about 1 μg/kg, about 10 μg/kg, about 25 μg/kg, about 50 μg/kg, about 75 μg/kg, about 100 μg/kg, about 125 μg/kg, about 150 μg/kg per day , about 175 μg/kg, about 200 μg/kg, about 225 μg/kg, about 250 μg/kg, about 275 μg/kg, about 300 μg/kg, about 325 μg/kg, about 350 μg/kg, about 375 μg/kg, about 400 μg/kg , about 425 μg/kg, about 450 μg/kg, about 475 μg/kg, about 500 μg/kg, about 525 μg/kg, about 550 μg/kg, about 575 μg/kg, about 600 μg/kg, about 625 μg/kg, about 650 μg/kg , about 675 μg/kg, about 700 μg/kg, about 725 μg/kg, about 750 μg/kg, about 775 μg/kg, about 800 μg/kg, about 825 μg/kg, about 850 μg/kg, about 875 μg/kg, about 900 μg/kg , about 925 μg/kg, about 950 μg/kg, about 975 μg/kg, about 1 mg/kg, about 5 mg/kg, about 10 mg/kg, about 15 mg/kg, about 20 mg/kg, about 25 mg/kg, about 30 mg/kg , about 35mg/kg, about 40mg/kg, about 45mg/kg, about 50mg/kg, about 60mg/kg, about 70mg/kg, about 80mg/kg, about 90mg/kg, about 100mg/kg, about 125mg/kg , about 150 mg/kg, about 175 mg/kg, about 200 mg/kg, or about 300 mg/kg of body weight.
The method of any one of claims 1-4, wherein the second agent is administered in an amount of about 0.005 mg/day to about 5000 mg/day, such as about 0.005, 0.05, 0.5, 5, 10, 20, 30, 40 ,50,100,150,200,250,300,350,400,450,500,550,600,650,700,750,800,850,900,950,1000,1500,2000,2500,3000,3500 , 4000, 4500 or 5000mg/day dose administration;

Preferably, the second agent is administered in an amount of about 5 mg/day.
The method of any one of claims 1-5, wherein the second agent is administered at about 1 ng/kg to about 200 mg/kg, about 1 μg/kg to about 100 mg/kg, or about 1 mg/kg to about 50 mg/kg per day. Administration in an amount of kg body weight, for example, about 1 μg/kg, about 10 μg/kg, about 25 μg/kg, about 50 μg/kg, about 75 μg/kg, about 100 μg/kg, about 125 μg/kg, about 150 μg/kg per day , about 175 μg/kg, about 200 μg/kg, about 225 μg/kg, about 250 μg/kg, about 275 μg/kg, about 300 μg/kg, about 325 μg/kg, about 350 μg/kg, about 375 μg/kg, about 400 μg/kg , about 425 μg/kg, about 450 μg/kg, about 475 μg/kg, about 500 μg/kg, about 525 μg/kg, about 550 μg/kg, about 575 μg/kg, about 600 μg/kg, about 625 μg/kg, about 650 μg/kg , about 675 μg/kg, about 700 μg/kg, about 725 μg/kg, about 750 μg/kg, about 775 μg/kg, about 800 μg/kg, about 825 μg/kg, about 850 μg/kg, about 875 μg/kg, about 900 μg/kg , about 925 μg/kg, about 950 μg/kg, about 975 μg/kg, about 1 mg/kg, about 5 mg/kg, about 10 mg/kg, about 15 mg/kg, about 20 mg/kg, about 25 mg/kg, about 30 mg/kg , about 35mg/kg, about 40mg/kg, about 45mg/kg, about 50mg/kg, about 60mg/kg, about 70mg/kg, about 80mg/kg, about 90mg/kg, about 100mg/kg, about 125mg/kg , about 150 mg/kg, about 175 mg/kg, about 200 mg/kg, or about 300 mg/kg of body weight.
The method of any one of claims 1-6, wherein the daily dose of the first agent and/or the second agent is administered in one dose or divided into two, three or four doses;

Preferably, the daily dose of the first medicament is administered in two divided doses and the daily dose of the second medicament is administered in one go.
The method of any one of claims 1-7, wherein the first agent and/or the second agent are administered continuously for at least 1 week, at least 2 weeks, at least 3 weeks, at least 4 weeks, at least 5 weeks, At least 6 weeks, at least 7 weeks, at least 8 weeks, at least 9 weeks, at least 10 weeks, at least 11 weeks, at least 12 weeks, at least 13 weeks, at least 14 weeks, at least 15 weeks, at least 16 weeks, at least 17 weeks, at least 18 weeks weeks, at least 19 weeks, at least 20 weeks, at least 21 weeks, at least 22 weeks, at least 23 weeks, at least 24 weeks, at least 25 weeks, at least 30 weeks, at least 35 weeks, at least 40 weeks, at least 45 weeks, at least 50 weeks, At least 51 weeks, at least 52 weeks, at least 53 weeks, at least 54 weeks, or at least 55 weeks.
The method of any one of claims 1-8, wherein one or more (eg 1, 2, 3, 4, 5, 6, 7, 8 , 9 or 10) courses of treatment, wherein each course of treatment lasts at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 8 days, at least 9 days, at least 10 days, at least 11 days, at least 12 days, at least 13 days, at least 14 days, at least 15 days, at least 16 days, at least 17 days, at least 18 days, at least 19 days, at least 20 days, at least 21 days, at least 22 days, at least 23 days, at least 24 days , at least 25 days, at least 30 days, at least 35 days, at least 40 days, at least 45 days, or at least 50 days; , 9, 10 days, two weeks, three weeks or four weeks.
The method according to any one of claims 1-9, wherein said first agent and/or said second agent are injected (such as intravenously, intraarterially, subcutaneously, intraperitoneally, intramuscularly, including drip) Or transdermally; or orally, buccally, nasally, transmucosally, topically, in the form of ophthalmic preparations or by inhalation.
The method of any one of claims 1-10, wherein the first medicament and/or the second medicament are selected from the group consisting of tablets, capsules, lozenges, hard candies, powders, sprays, creams , ointments, suppositories, gels, pastes, lotions, ointments, aqueous suspensions, injectable solutions, elixirs and syrups.
The method of any one of claims 1-11, wherein the method improves the following pathophysiological manifestations in the individual: cognitive behavioral abnormalities, neurodegenerative changes (such as progressive synapse/neuron loss and brain atrophy), beta -Amyloid deposition, abnormal phosphorylation of Tau and its resulting neurofibrillary tangles, glial cell activation and inflammation and/or impaired glucose metabolism in the brain.
The method of any one of claims 1-12, wherein the second medicament is administered in a lower amount than when the second medicament is administered alone or when the first medicament is not administered. dosage.
The method of any one of claims 1-13, wherein the first agent enhances the efficacy of the second agent in the treatment of a neurodegenerative disease and/or reduces the efficacy of the second agent in the treatment of a neurodegenerative disease side effects.
14. The method of any one of claims 1-14, wherein said individual has received monotherapy with said second agent prior to receiving the combined administration of said first agent and said second agent.
The method of any one of claims 1-15, wherein the individual is a non-APOE ε4 allele carrier; preferably, the individual is a mild to moderate Alzheimer's disease patient who does not carry the APOE ε4 allele sick individual.
The method of any one of claims 1-16, wherein the method results in improvement or no deterioration or a reduced rate of deterioration in at least one of the assessments selected from: Alzheimer's Disease Assessment Scale - Cognitive Component Table (ADAS-cog); Clinical Dementia Rating Pencil Box (CDR-sb); Alzheimer's Disease Collaborative Study Activities of Daily Living Scale (ADCS-ADL); Neuropsychiatric Inventory (NPI); MMSE);

Preferably, the method results in a reduction in the rate of deterioration in the ADAS-cog score;

More preferably, the method reduces the rate of deterioration of the ADAS-cog score by an average of 1-5 points, preferably 2-3.5 points.