CN109953991B - Use of gelsmin for treating diseases or disorders associated with amyloid deposition and/or tauopathy - Google Patents

Abstract

The present invention relates to the use of gelsemine for the treatment of diseases or disorders associated with amyloid deposition and/or tauopathy.

Description

Use of gelsmin for treating diseases or disorders associated with amyloid deposition and/or tauopathy

Technical Field

The invention relates to a new application of gelsemium elegans (koumine) or a pharmaceutically acceptable salt thereof as a gelsemium alkaloid monomer, in particular to an application of gelsemium elegans or a pharmaceutically acceptable salt thereof in preparing a medicament for treating diseases or symptoms related to amyloid deposition and/or tauopathy, and particularly Alzheimer disease.

Background

"amyloidosis" is a condition characterized by the accumulation of various insoluble fibrillar proteins (amyloid) in the tissues of a patient. Amyloid deposits are formed by the accumulation of amyloid protein followed by further pooling of the accumulation and/or amyloid protein. The accumulation of amyloid in living tissue is a cause or major factor in the pathology of a variety of so-called amyloid diseases (e.g., alzheimer's disease, parkinson's disease, huntington's disease, and prion diseases). Certain types of amyloidosis can occur primarily in the central nervous system, such as aggregation of amyloid beta-protein (a β) in alzheimer's disease, aggregation of tau protein in progressive supranuclear palsy, aggregation of alpha synuclein in parkinson's disease, aggregation of huntingtin in huntington's disease, and aggregation of prion proteins in creutzfeldt-jakob disease and other prion diseases. Amyloid beta is also known as amyloid beta peptide (a β). Other types of amyloidosis are systemic in nature, such as the accumulation of transthyretin in senile systemic amyloidosis.

Alzheimer's Disease (AD), also known as senile dementia, is a common degenerative disease of the central nervous system, and is clinically manifested by progressive memory impairment, cognitive dysfunction, personality changes, and other symptoms. The main pathological features of AD include: senile Plaques (SP) formed by aggregation of Abeta, neurofibrillary tangles (NFT) formed by abnormal aggregation of tau protein and the like appear in cerebral cortex and hippocampus to prevent and treat senile plaques (NFT)And a decrease in nerve cells in the cortex and hippocampus. The theory of microtubule-associated protein tau abnormality is that tau protein has the function of synthesizing and stabilizing neuronal cells, and the total amount of this protein is significantly increased in senile dementia patients and is dominated by abnormal hyperphosphorylation. Hyperphosphorylated tau protein pairs with other threadlike structures of tau and finally forms neurofibrillary tangles in the nerve cell body. When this occurs, the microtubules disintegrate, disrupting the transport system of the neuron. This may lead to a malfunction in biochemical communication between neurons, which in turn leads to cell death. The abeta toxicity theory states that the disease is an amyloidosis, which results from abnormal hydrolysis and misfolding of the intramembranous Amyloid Precursor Protein (APP). There is increasing evidence that a β plays a dominant role in the development and progression of AD. The neurotoxicity of A.beta.involves complex molecular mechanisms mainly involving the destruction of intracellular Ca²⁺Steady state, promoting formation of free radicals, and lowering K⁺The function of the channel, the enhancement of inflammatory reaction caused by inflammatory cytokines and the like. Abnormal deposition of a β is currently considered to be one of the major pathogenesis of alzheimer's disease.

In addition, studies have shown that Amyloid Spheroids (ASPD), a unique a β aggregate that does not exhibit toxicity to non-neuronal cells and naive neuronal cells but selectively causes death of functionally mature neuronal cells, plays an important role in the irreversible phase of alzheimer's disease onset. In addition, native amyloid spheres have also been detected in the brain of patients with dementia with lewy bodies, and thus amyloid spheres are considered to play an important role in the onset of dementia with lewy bodies.

Amyloid is also implicated in pathological changes in the nervous system of parkinson's disease. For example, lipofuscin, amyloid, filaments, etc. are deposited in and around neurons, causing their function to decline until loss, which can ultimately impair recent memory in patients, progressing to dementia. In addition, amyloid and some irregular, loosely arranged neurons together constitute senile plaques. Mutations in the gene encoding tau protein cause parkinsonism, which is pathologically characterized by the presence of a large number of neurofibrillary tangles in brain tissue. It is thought that cognitive impairment in parkinson's disease may be associated with the combined effects of alpha-synuclein, tau protein and a β. Alzheimer's disease and Parkinson's disease and the like are also called Amyloid Diseases (Amyloid Diseases).

Similarly, cerebral pathological examination of Cerebral Amyloid Angiopathy (CAA) patients often has senile changes such as Senile Plaques (SP) and neurofibrillary tangles (NFT), which are sometimes difficult to distinguish from AD. CAA is a disorder characterized by amyloid deposits in the vessel walls of the central nervous system, particularly in leptomeningeal and cortical arteries. CAA occurs primarily as a sporadic condition in the elderly, with incidence associated with increasing age. These dispersed CAA cases are attributed to the deposition of a β. The genetic form of CAA is usually familial, more severe and occurs earlier than dispersible CAA. More recently, CAA has also been considered as a potential contributor to the development of AD.

Amyloid deposition and/or tauopathies may also be characteristic of a variety of neurodegenerative or other disorders.

Gelsemium elegans Benth is an indole alkaloid found in Gelsemium elegans Benth, and is a monomer with the highest content and lower toxicity in Gelsemium elegans total alkaloids. Chinese patent CN 101323618B discloses a method for extracting and separating koumine from gelsemium elegans. The inventors of the present application have found a novel use of koumine in diseases or disorders associated with amyloid deposition and/or tauopathy, in particular alzheimer's disease.

Disclosure of Invention

In one aspect of the invention, there is provided the use of koumine or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for the treatment of a disease or disorder associated with amyloid deposition and/or tauopathy in a subject in need thereof.

In another aspect of the present invention, there is provided the use of koumine or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for reducing amyloid, e.g. inhibiting or reducing amyloid production and/or deposition, in a subject in need thereof.

In a further aspect of the invention there is provided the use of koumine or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for inhibiting or reducing neurofibrillary tangles in an individual in need thereof.

In a further aspect of the invention there is provided the use of koumine or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for the treatment or alleviation of cognitive dysfunction in an individual in need thereof.

Brief description of the drawings

FIG. 1 shows the effect of gelsemin on learning acquisition ability of Alzheimer's disease-like rats. The abscissa in the graph represents time (days), and the ordinate represents escape latency (seconds); ctrl: a control group; OA: okadaic acid; KMH: gelsemiq Elegano hydrochloride.^###P is less than 0.001, compared with the control group;^*P＜0.05，^***p is less than 0.001, compared with the model group.

FIG. 2 shows the effect of gelsemine on spatial memory in Alzheimer's disease-like rats. The ordinate in the figure represents the times of crossing the position area of the original platform; ctrl: a control group; OA: okadaic acid; KMH: gelsemiq Elegano hydrochloride.^###P is less than 0.001, compared with the control group;^*P＜0.05，^***p is less than 0.001, compared with the model group.

FIG. 3 shows the effect of gelsemium on pathological changes in hippocampal tissue in Alzheimer's disease-like rats. Ctrl: a control group; OA: okadaic acid; KMH: koumine hydrochloride; CA: hippocampal horn; DG: and (6) tooth-shaped back.

Detailed Description

The present invention will now be described in detail with reference to representative embodiments thereof. These embodiments are merely illustrative and should not be construed as limiting the scope of the invention in any way. On the contrary, the invention is intended to cover all alternatives, modifications and equivalents that may be included within the scope of the invention as defined by the appended claims.

Unless defined otherwise, technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

The term "koumine" as used herein has the following structural formula, formula C₂₀H₂₂N₂O, which is an alkaloid monomer from gelsemium elegans, may also be synthesizedObtaining:

as used herein, pharmaceutically acceptable salts of koumine include salts of koumine with organic or inorganic acids. Exemplary pharmaceutically acceptable salts include, but are not limited to: hydrochloride, sulfate, hydrobromide, hydroiodide, nitrate, bisulfate, phosphate, acid phosphate, citrate, acetate, oxalate, isonicotinate, lactate, salicylate, acid citrate, tartrate, oleate, tannate, pantothenate, bitartrate, ascorbate, succinate, maleate, gentisate, fumarate, gluconate, glucuronate, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate and pamoate. Preferably, the pharmaceutically acceptable salt is koumine hydrochloride.

As used herein, "pharmaceutically acceptable" means that the compound, composition or carrier is suitable for administration to a subject to achieve the use or method described herein without undue adverse side effects.

Pharmacological experiments show that the gelsemium can reduce clinical main symptoms such as cognitive impairment of Alzheimer disease and the like in a dose-dependent manner, and obviously inhibit and reverse main characteristic pathological changes such as senile plaques and neurofibrillary tangles.

Accordingly, in some embodiments, the present invention provides the use of koumine or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for the treatment of a disease or disorder associated with amyloid deposition and/or tauopathy in a subject in need thereof.

In other embodiments, the present invention provides a method of treating a disease or disorder associated with amyloid deposition and/or tauopathy in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of gelsemine or a medicament comprising a therapeutically effective amount of gelsemine.

The term "subject" refers to a mammal, including, but not limited to, primates (e.g., humans, monkeys, chimpanzees, gorillas, etc.), rodents (e.g., rats, mice, gerbils, hamsters, ferrets, and the like), lagomorphs, porcines (e.g., pigs, piglets), equines, canines, felines, etc., preferably humans.

The term "treatment" refers to a complete or partial cure of a disease, including but not limited to one, or a combination of two or more, selected from the following: reducing or eliminating the etiology of the disease or disorder; ameliorating or eliminating pathological changes thereof; alleviating or eliminating one or more symptoms thereof; slowing or arresting its progression; lessening its severity; reducing the incidence rate; reducing the recurrence thereof; and improving the prognosis thereof.

An "amyloid deposition-associated disease or disorder" is a disease or disorder associated with amyloid deposition, which may occur in the Central Nervous System (CNS) or outside the CNS. In some embodiments, the amyloid protein is selected from the group consisting of amyloid beta, amylin, amyloid a protein, serum amyloid a, alpha-synuclein, Sc-type prion protein (PrP)^Sc) Huntingtin, calcitonin, atrial natriuretic peptide, transthyretin, lysozyme, beta 2-microglobulin, calcium binding microfilamentin, cystatin and amyloid light chain protein (AL), preferably beta amyloid. In some embodiments, the amyloid deposition involves amyloid beta, amylin, amyloid A protein, serum amyloid A, alpha-synuclein, Sc-type prion protein (PrP)^Sc) Huntingtin, calcitonin, atrial natriuretic peptide, transthyretin, lysozyme, beta 2-microglobulin, calcium binding microfilamentin, cystatin and/or amyloid light chain protein (AL), preferably amyloid beta. The amyloid deposition-related disease or disorder may include, but is not limited to: alzheimer's disease, including early-onset alzheimer's disease, late-onset alzheimer's disease, and presymptomatic alzheimer's disease; parkinson's disease, including postencephalitic parkinson's disease; huntington's disease; multiple functionA myeloma; amyloid polyneuropathy; systemic neuritic amyloidosis, amyloid cardiomyopathy; systemic senile amyloidosis; hereditary cerebral hemorrhage with amyloidosis; down syndrome; scrapie of sheep; medullary thyroid carcinoma; isolated atrial amyloidosis; dialyzing the patient for beta 2-microglobulin amyloidosis; inclusion body myositis; prion diseases including kuru, creutzfeldt-jakob disease, gerstmann-Straussler-Scheinkersyndrome, fatal familial insomnia; beta 2-amyloid deposition in muscle wasting diseases; type II diabetes; cognitive dysfunction in diabetes; familial mediterranean fever; moore-weidi syndrome; isolated cardiac amyloidosis; islet amyloid polypeptide (amylin); insulinomas; familial amyloidosis; familial amyloid polyneuropathy; familial amyloid nephropathy with urticaria; familial amyloidosis, cardiomyopathy; dementia with lewy bodies; fibrinogen-associated amyloidosis; bovine spongiform encephalopathy; progressive supranuclear palsy; frontotemporal dementia (FTD); frontotemporal lobar degeneration disease (FTLD); british/denmark dementia; SAA amyloidosis; amyotrophic Lateral Sclerosis (ALS); spinocerebellar ataxia (SCA1, SCA3, SCA6, or SCA 7); atrophy of lower nucleus of thalamus of dentalis pallidus; spinal bulbar muscular atrophy; amyloid cerebrovascular disease, including hereditary amyloid cerebrovascular disease; demyelinating disease; dementia due to hypothyroidism; toxic dementia, including acute and chronic alcoholism; normal intracranial hydrocephalus; traumatic brain dementia; aids dementia syndrome; and paralytic dementia.

A "tauopathy-associated disease or disorder" is a neurodegenerative disease involving tauopathy. In a preferred embodiment, the tauopathy is a neurofibrillary tangle. The tauopathy-related diseases or disorders include, but are not limited to: alzheimer's disease; vascular dementia; amyotrophic lateral sclerosis; dementia with Parkinson's disease; dementia with silvery particles; corticobasal degeneration; Creutzfeldt-Jakob disease; dementia pugilistica; diffuse neurofibrillary tangles with calcification; down syndrome; frontotemporal dementia, including frontotemporal dementia with chromosome 17-associated parkinson's disease; gertman-starchler-schyneck syndrome; Harlervorden-Spatz disease (Hallervorden-Spatz disease); myotonic dystrophy; niemann Pick disease type C (Niemann-Pick disease type C); non-synaptonemal motor neuron disease with neurofibrillary tangles; pick's disease; postencephalitic parkinsonism; prion cerebral amyloid angiopathy; progressive subcortical hyperplasia; progressive supranuclear palsy; subacute sclerosing panencephalitis; dementia of the neuro-entanglement type; cognitive dysfunction in diabetes; toxic dementia, including acute and chronic alcoholism; and hepatolenticular degeneration. Some of these diseases also include a β deposition. For example, alzheimer's disease, vascular dementia, diabetic cognitive dysfunction, alcoholism, hepatolenticular degeneration with concurrent a β deposition and tauopathy. Similarly, prion-mediated diseases such as Creutzfeldt-Jakob disease, prion cerebral amyloid angiopathy, and Gettmann-Straussler-Schenk syndrome, as well as normal intracranial pressure hydrocephalus, traumatic brain dementia, AIDS dementia complex, and paralytic dementia also have tauopathies.

Preferably, the amyloid deposition and/or tauopathy-associated disease or disorder is selected from: alzheimer's disease, vascular dementia, Parkinson's disease, Huntington's disease, Down's syndrome, Lewy body dementia, progressive supranuclear palsy, frontotemporal dementia, Parkinson's disease dementia, amyloid cerebrovascular disease, dementia of the neuro-entanglement type, Creutzfeldt-Jakob disease, and combinations thereof. Particularly preferably, the disease or disorder is alzheimer's disease.

In some embodiments, the present invention provides the use of koumine or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for reducing amyloid, e.g. inhibiting or reducing amyloid production and/or deposition, in a subject in need thereof. In some embodiments, the amyloid protein is selected from the group consisting of amyloid beta, amylin, amyloid a protein, serum amyloid a, alpha-synuclein, Sc-type prion protein (PrP)^Sc) Huntingtin, calcitonin, atrial natriuretic peptide, transthyretin, lysozyme, beta 2-microglobulin,Calcium binds to microfilamentin, cystatin and amyloid light chain protein (AL), preferably amyloid beta. In some embodiments, the amyloid deposition involves amyloid beta, amylin, amyloid A protein, serum amyloid A, alpha-synuclein, Sc-type prion protein (PrP)^Sc) Huntingtin, calcitonin, atrial natriuretic peptide, transthyretin, lysozyme, beta 2-microglobulin, calcium binding microfilamentin, cystatin and/or amyloid light chain protein (AL), preferably amyloid beta.

In other embodiments, the present invention provides the use of koumine or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for inhibiting or reducing neurofibrillary tangles in a subject in need thereof.

In some embodiments, the present invention provides a method of reducing amyloid, e.g., inhibiting or reducing amyloid production and/or deposition, in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of gelsemin or a pharmaceutically acceptable salt thereof, or a therapeutically effective amount of a medicament comprising gelsemin or a pharmaceutically acceptable salt thereof.

In other embodiments, the present invention provides a method for inhibiting or reducing neurofibrillary tangles in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of gelsemine or a pharmaceutically acceptable salt thereof, or a therapeutically effective amount of a medicament comprising gelsemine or a pharmaceutically acceptable salt thereof.

In some embodiments, the present invention provides the use of koumine or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for the treatment or alleviation of cognitive dysfunction in an individual in need thereof.

In other embodiments, the present invention provides a method for treating or reducing cognitive dysfunction in an individual in need thereof, comprising administering to the individual a therapeutically effective amount of koumine or a pharmaceutically acceptable salt thereof, or a therapeutically effective amount of a medicament comprising koumine or a pharmaceutically acceptable salt thereof.

Cognitive function is a psychological or intellectual process involving neurological or symbolic operations, including, but not limited to, communication, insight, understanding, reasoning, learning, memory, thinking, awareness, concentration, attention, alertness, motivation, drawing conclusions, performing functions, establishing imagery, and judgment ability. In animal model systems, cognitive function can be measured by a variety of conventional means known in the art, including the use of the Morris water maze, the Barnes circular maze, the elevated radial arm maze, or the T-maze. Other tests known in the art may also be used to assess cognitive function, such as new object recognition and scent recognition tasks. In a preferred embodiment, the cognitive dysfunction is selected from the group consisting of: learning disorders, memory disorders, disorientation, language dysfunction, visuospatial dysfunction, personality disorders, affective disorders, cognitive function impairment, and dementia. Cognitive dysfunction is a defect or abnormality in cognitive function as described above, including learning and memory disorders. In some embodiments, the cognitive dysfunction is a cognitive dysfunction in a disease or condition selected from the group consisting of:

alzheimer's disease, including early-onset alzheimer's disease, late-onset alzheimer's disease, and presymptomatic alzheimer's disease; parkinson's disease, including postencephalitic parkinson's disease; huntington's disease; hereditary cerebral hemorrhage with amyloidosis; down syndrome; prion diseases including kuru, creutzfeldt-jakob disease, gerstmann-straussler-scheinker syndrome, fatal familial insomnia; cognitive dysfunction in diabetes; familial amyloid polyneuropathy; bovine spongiform encephalopathy; progressive supranuclear palsy; vascular dementia, including cerebral ischemic dementia, cerebral hemorrhagic dementia, and hypoperfusion dementia, such as multi-infarct dementia, lacunar state, and Binswanger's disease; dementia with lewy bodies; frontotemporal dementia; frontotemporal lobar degeneration; british/denmark dementia; spinocerebellar ataxia (SCA1, SCA3, SCA6, or SCA 7); atrophy of lower nucleus of thalamus of dentalis pallidus; amyloid cerebrovascular disease, including hereditary amyloid cerebrovascular disease; dementia with Parkinson's disease; dementia with silvery particles; corticobasal degeneration; dementia pugilistica; diffuse neurofibrillary tangles with calcification; hallowden-spastus disease; niemann pick disease type C; pick's disease; prion cerebral amyloid angiopathy; progressive subcortical hyperplasia; subacute sclerosing encephalitis; dementia of the neuro-entanglement type; normal intracranial hydrocephalus; traumatic brain dementia; demyelinating disease; dementia due to hypothyroidism; dementia due to vitamin deficiency; toxic dementia, including acute or chronic alcoholism; dementia due to infectious diseases, including aids dementia complex and paralytic dementia; hepatolenticular degeneration; stroke; a paraneoplastic syndrome; mitochondrial encephalomyopathy; degeneration of the spinal cord and cerebellum; primary basal ganglia calcification; dementia due to depression and other psychiatric disorders; multiple sclerosis; HIV encephalopathy; brain abscess; toxic and metabolic encephalopathies, including heroin encephalopathy, hepatic encephalopathy, and uremic encephalopathy; amyotrophic lateral sclerosis dementia; degeneration of globus pallidus substantia nigra pigment; neurosyphilis; leptospirosis of nerve endings; lyme disease; viral encephalitis; after fungal and bacterial meningitis/encephalitis; progressive multifocal leukoencephalopathy; and combinations thereof.

The medicament comprises the koumine or a pharmaceutically acceptable salt thereof as the only active ingredient, or the combination of the koumine or the pharmaceutically acceptable salt thereof and one or more other active ingredients, and an optional pharmaceutically acceptable carrier. In some embodiments, the medicament comprises only koumine as an active ingredient. In other embodiments, the medicament comprises koumine and one or more additional active ingredients; in such embodiments, koumine or a pharmaceutically acceptable salt thereof and one or more other active ingredients may be formulated separately for inclusion in separate compositions or formulated together in a single composition comprising them.

In some embodiments of the methods of the invention, koumine or a pharmaceutically acceptable salt thereof is administered to the individual as the sole active ingredient, or in combination with one or more other active ingredients. In the case of combination therapy, koumine or a pharmaceutically acceptable salt thereof and one or more other active ingredients may be administered to the individual simultaneously or sequentially. The combined administration includes the use of separate compositions containing gelsemium or a pharmaceutically acceptable salt thereof and one or more other active ingredients, respectively, or a single composition containing the above active ingredients, administered simultaneously or sequentially in any order.

The one or more physiologically active ingredients thereof are one or more chemotherapeutic agents, biotherapeutic agents, or combinations thereof known in the art to have therapeutic efficacy for the above-mentioned diseases or conditions. By using in combination with gelsemine, effects including, but not limited to, the following can be achieved: combination therapy to enhance the overall efficacy of each drug component; lowering the dosage of other pharmacologically active ingredients to avoid side effects; and maintaining therapeutic efficacy.

The one or more further active ingredients may be selected from: a cholinesterase inhibitor; cannabinoids; a statin-based drug; a beta secretase inhibitor; gamma secretase inhibitors; an a β aggregation inhibitor; a β -peptide specific antibody; a selective estrogen receptor molecule; an antihypertensive agent; an anti-inflammatory agent; an antioxidant; and combinations thereof.

Cholinesterase inhibitors include, but are not limited to, tacrine hydrochloride, donepezil, rivastigmine, or galantamine.

The cannabinoids may be exogenous cannabinoids and endogenous cannabinoids. The endocannabinoid may be, for example, anandamide, tetrahydrocannabinol, 2-arachidonic acid glycerol ether, N-arachidonic acid-dopamine or Virodham amine.

The statin may be, for example, atorvastatin, rosuvastatin, fluvastatin, mevastatin, pitavastatin, pravastatin, rosuvastatin, and simvastatin.

Beta-secretase cleaves the first amino acid at the N-terminus of the APP molecule and is the rate-limiting enzyme for a β production. Beta-secretase inhibitors include, for example, CTS-21166, NB-53, KMI-429 and GSK188909, GRL-8234, TAK-070, and rosiglitazone.

The gamma secretase performs the last step of cleavage of a β production, determining the length of a β, and plays a key role in regulating the ratio of a β 40 to a β 42. Semagachtat (LY-450139) is a gamma secretase inhibitor. Other gamma secretase inhibitors are BMS-708163, R-flurbiprofen (Tarenflurbil) and NIC 5-15 (Huyman corporation).

Examples of A.beta.aggregation inhibitors include homotaurine (3-amino-1-propanesulfonic acid, Tramiprosate or NC-531), Scyllo-inositol (Scyllo-inositol or ELN D005), and metal ion chelators including Cu²⁺、Zn²⁺Or Fe³⁺Chelating agents, such as clioquinol and PBT 2.

Examples of a β -peptide specific antibodies include: baclizumab (bapineuzumab) (Elan/qiangsheng), sorafezumab (solarezumab) (LY2062430) (lilies), gamma globulin iv (baxter), and PF-4360365 (fevere).

The Selective Estrogen Receptor Molecule (SERM) may be, for example, raloxifene.

Antihypertensive agents include alpha-blockers, beta-blockers, alpha, beta-blockers, angiotensin converting enzyme inhibitors, angiotensin receptor blockers (e.g., valsartan), calcium channel blockers and diuretics.

Anti-inflammatory agents useful in the present invention include, but are not limited to:

mast cell stabilizers such as cromolyn, cromolyn derivatives, cromolyn analogs, eugenol, nedocromil, perolescent, olopatadine, fraxinin G1, framycetin B1, fraxinin M1, deoxynivalenol (deoxynivalenol), zearalenone, ochratoxin a, fumonisin B1, hydrolyzed fumonisin B1, patulin, and ergotamine;

steroidal anti-inflammatory agents, such as prednisone;

non-steroidal anti-inflammatory agents (NSAIDs) such as acetylsalicylic acid, diflunisal, salsalate, ibuprofen, dexibuprofen, naproxen, fenoprofen, indoprofen, dexketoprofen, flurbiprofen, oxaprozin, loxoprofen, indomethacin, tolmetin, sulindac, etodolac, ketorolac, diclofenac, nabumetone, piroxicam, meloxicam, tenoxicam, droxicam, lornoxicam, isoxicam, mefenamic acid, meclofenamic acid, flufenamic acid, celecoxib, lixilone and hyperforin;

antioxidants, such as vitamin E or selegiline, can also improve cognitive function, mood, and/or social behavior in an individual. Other antioxidants useful in the present invention include: garlic extract, curcumin, melatonin, resveratrol, ginkgo biloba extract, green tea extract, vitamin C and the like.

The medicament also optionally comprises pharmaceutically acceptable carriers such as excipients, disintegrants, diluents, binders, glidants, lubricants, pH adjusting agents, preservatives, dispersants, suspending agents, ointment bases, emulsifiers, emollients, penetration enhancers, surfactants, propellants, flavoring agents, sweeteners, drug release modifiers, and the like. One skilled in the art can select an appropriate carrier depending on the desired formulation form, mode of administration, drug release characteristics, and the like of the drug.

The medicament may be formulated into dosage forms in solid, semi-solid, or liquid form suitable for administration by any of the recognized modes of administration available in the art, including but not limited to: (1) suitable for oral administration: such as tablets, capsules, powders, granules, troches, aqueous or anhydrous solutions or suspensions, syrups, and the like; (2) suitable for parenteral administration: e.g., subcutaneous, intramuscular, or intravenous injection, such as a sterile solution or suspension; (3) suitable for topical administration: for example, a plaster, ointment, cream, spray, gel, or the like for application to the skin or mucosa; (4) suitable for transdermal administration: such as a patch, gel paste, etc.; (5) suitable for vaginal or rectal administration: such as suppositories, creams, gels or effervescent tablets, etc.

As used herein, the term "therapeutically effective amount" refers to the amount of active ingredient that will achieve the desired therapeutic efficacy of the use or method when administered.

In the uses or methods described herein, the dosage of the koumine or pharmaceutically acceptable salt thereof and/or the one or more other active ingredients will generally depend on a variety of factors, including the severity of the individual, disorder or condition being treated, the rate of administration and the judgment of the prescribing physician. Generally, an effective dose is from about 0.0001 to about 50mg per kg body weight per day, e.g., from about 0.0005 to about 25 mg/kg/day or from about 0.001 to about 5 mg/kg/day (single or divided administration). For a 70kg human, this may amount to about 0.007 mg/day to about 3500 mg/day, for example about 0.035 mg/day to about 1750 mg/day or about 0.07 mg/day to about 350 mg/day. In some cases, dosage levels not higher than the lower limit of the aforesaid range may be sufficient, while in other cases still larger doses may be employed without causing any harmful side effects, and if necessary, the larger doses may be divided into several smaller doses for administration throughout the day.

The amount of gelsemium or a pharmaceutically acceptable salt thereof and/or the one or more other active ingredients in the medicament or in the method may be from about 0.001mg to about 500mg, such as from about 0.01 to 200mg, from about 0.1 to 20mg or from about 0.5 to 10mg, preferably from about 0.1 to 20mg, such as from about 0.1mg, 0.5mg, 1mg, 2mg, 3mg, 4mg, 5mg, 6mg, 8mg, 10mg, 12mg, 15mg or about 20mg etc.

The present invention is described in more detail below by way of examples, which are not intended to limit the invention in any way, but the scope of the invention is defined only by the claims.

Examples

Example 1:

the Okadaic Acid (OA) induced Alzheimer disease model can simulate the main clinical manifestations of Alzheimer disease, namely cognitive dysfunction and the main pathological features, namely senile plaques and neurofibrillary tangles, and is a model suitable for screening Alzheimer disease treatment drugs. In this example, the therapeutic effect of gelsemium on alzheimer's disease was studied on a rat model of alzheimer's disease using the learning and memory ability (learning and acquisition ability and spatial memory ability) and the pathological changes of hippocampal tissue as indices.

1. Material

1.1 animals

Healthy male Sprague-Dawley rats, clean grade, body mass 160-: SCXK (min) 2012 and 0001 ]. The adaptive breeding is carried out for 5 days before the experiment, the experiment is started when the body mass reaches 220g, the breeding is carried out in the environment with the light-dark period of 12/12h (the illumination time is 8: 00-20: 00), the constant temperature (23 +/-2 ℃) and the constant humidity (50 +/-5%) and the diet is freely drunk. The experimental procedures followed the regulations promulgated by the international and local committees for the use and protection of laboratory animals.

1.2 drugs and reagents

Koumine hydrochloride (KMH) was obtained by extraction and purification from wild gelsmium elegans plant produced in Fujia by the applicant, lab batch No. 201503 (purity 99.9% by hplc). Okadaic acid was purchased from sigma, usa.

1.3 Experimental instruments

The digital display type brain stereotaxic apparatus, the KDS310 type micro-injection pump, the 78001 micro hand-held skull drill, the micro sample injector (1 mu l flat head), the R510-25 type anesthesia machine air pump, the 68623 type anesthesia mask for the stereotaxic apparatus-rat, the V101 type anesthesia induction box-rat, and are purchased from Shenzhen Riword Life technologies, Ltd. Model RD1101 Morris water maze available from shanghai mobile number information technology ltd. Small animal anesthesia machine, available from MIDMARK corporation, usa. A TP1020 type semi-automatic dehydrator, an EG1150 type full-automatic paraffin embedding machine, and an RM2235 type full-automatic paraffin embedding machine, which are available from Leica corporation. Model 80i biomicroscope available from Nicon corporation.

2. Method of producing a composite material

After 5 days of adaptive feeding, floating, not very swimming rats and rats with too short or too long mean escape latency were removed by four-round per day Morris water maze primary screening. The ventriculo-cerebri catheter was implanted (day 1 of surgery), and was intraperitoneally injected with penicillin sodium for 3 consecutive days after surgery to resist infection. Rats were randomly divided into control group, model group, gelsemium high, medium and low dose treatment groups. On the 3 rd day after embedding the lateral ventricle catheter, injecting 0.4mM OA into the lateral ventricle of the rats of the model group and the gelsmin treatment group, 1 mul each time, and 1 time every 2 days for 4 times in total to establish an Alzheimer disease model of the rats; the control group was injected with an equal volume of vehicle. On the next day after last injection of OA into lateral ventricle, the gelsemium treatment group is administered with gelsemium hydrochloride solution of 0.056, 0.28 or 1.4mg/kg by intragastric administration for 1 time and 10 consecutive days; the other groups were gavaged with equal volumes of vehicle.

And (3) observation indexes are as follows:

(1) effect of Gelsemiun on learning and memory disorders in Alzheimer's disease-like rats

Influence of koumine on learning and acquisition ability of Alzheimer disease-like rats

After the last injection in the lateral ventricle for 2 weeks, the rats were examined for their ability to navigate directionally by the Morris method. Rats were placed in a water tank without a platform for 90s the day before the experiment, so that they were adapted to the maze environment and swim. The experiment is carried out for 5 days, and the positioning navigation test is carried out for the first 4 days: in order to test the learning and memory acquiring capacity of the rat, the rat is placed into water from four water entry points facing the pool wall, and the time required by the rat from entering the water to climbing the platform within 90s is recorded, namely the escape latency is recorded. If the rat could not find the platform within 90s, the experimenter uses a rod to guide the platform and stays the platform for 10s, and then the platform is put back into the cage, and the escape latency is recorded as 90 s. After each test, the rats were quickly dried with a dryer. Each rat was trained four times a day, i.e. from four different entry points into water each time, and the average was taken as the escape latency of the rat per day.

(ii) influence of Gelsemii Elegantis on spatial memory of Alzheimer's disease-like rats

The spatial exploration ability of rats was examined by the Morris method. The space exploration test was performed on day 5 of the Morris experiment: in order to test the reproducibility of the learning and memory of the rat, the platform is removed, the rat is placed into the water pool from the farthest water entry point to the pool wall, and the number of times of crossing the position area of the original platform within 120s is recorded.

(2) Effect of Gelsemii Georgi on pathological changes of hippocampal tissue of Alzheimer-like rats

After the behavioral experiment is finished, the hippocampus of rats in the control group, the model group and the gelsemium medium-dose treatment group is taken, the tissue chemical staining is carried out by a Bielschowsky staining method, and the pathological change condition of the hippocampus tissue of each group of rats is observed under a microscope.

3. Results

3.1 Effect of Gelsemiin on learning and memory disorder of Alzheimer's disease-like rats

Firstly, the effect of koumine on learning and acquiring capacity of rats with Alzheimer disease

The results of the directional voyage experiment are shown in figure 1. There was no significant difference in mean escape latency for groups of rats prior to molding. After modeling, the escape latency of the rats in the model group is stably maintained at a higher level, and the escape latency of the rats in the control group is progressively reduced from the 1 st day of training; the escape latency of the rats in the gelsemin treatment group is also reduced progressively, the level is between that of the model group and that of the control group, wherein the effect of the dose group (0.28mg/kg) in the gelsemin is more obvious, and the gelsemin has an improvement effect on the symptoms of low learning and acquisition capacity of the Alzheimer disease.

② the action of koumine on Alzheimer's disease-like rat space memory ability

The results of the space exploration experiment are shown in figure 2. The number of times that the rats in the control group pass through the original platform is the largest, the model group is obviously less than that of the control group, the gelsemium treatment group is between the control group and the model group, and the curative effect of the dose group in the gelsemium is the most obvious, which shows that the gelsemium has an improvement effect on the symptoms of low spatial memory capacity of the Alzheimer disease.

The result of the space exploration experiment is consistent with the result of the directional navigation experiment, and the koumine is prompted to have the effect of improving the learning and memory disorder of the Alzheimer disease.

3.2 Effect of Gelsemiin on pathological changes in Hippocampus tissue of Alzheimer-like rats

After the behavioral experiment is finished, taking the hippocampus of rats in the control group, the model group and the gelsemium medium-dose treatment group, and observing the pathological changes of the hippocampus tissues of the rats in each group by a Bielschowsky staining method. Pathological results of hippocampal CA1, CA2, CA3 region and DG are shown in fig. 3. Through 400 times magnification observation, the rat hippocampus of the control group is slightly stained, and the phenomena of senile plaques and neurofibrillary tangles are not seen. In the model group, the pyramidal neurons in each region of the hippocampus of the rat have obvious deep color, particularly a CA1 region and a DG, and neurofibrillary tangles occur in cells; senile plaques were visible outside cells in the CA3 region. The above lesions were reduced in the gelsemin treated rats, and the intracellular neurofibrillary tangles in each region and extracellular senile plaques in the CA3 region were significantly reduced. The results indicate that the gelsemin treatment group has obvious inhibition effect on the typical pathological changes of Alzheimer's disease, namely senile plaque and neurofibrillary tangle formation.

In conclusion, the koumine has an improvement effect on learning and memory disorder and characteristic pathological changes of main clinical symptoms of the Alzheimer disease, and the koumine has an effect of treating the Alzheimer disease.

All publications and patent applications cited in this application are indicative of the level of skill in the art to which this disclosure pertains. All of which are incorporated herein by reference in their entirety.

From the above description, one skilled in the art can ascertain the essential characteristics of this invention, and without departing from the spirit and scope thereof, can make various changes and modifications of the invention to adapt it to various usages and conditions. All such modifications and adaptations are intended to be included within the scope of the present invention. Accordingly, the scope of the invention should be determined by the appended claims and their legal equivalents.

Claims (17)

1. Use of koumine having the structure:

2. use according to claim 1, wherein said alzheimer's disease is selected from the group consisting of early-onset alzheimer's disease, late-onset alzheimer's disease and presymptomatic alzheimer's disease.

3. The use according to claim 1, wherein said medicament reduces amyloid-beta in said subject.

4. Use according to claim 1, wherein the medicament inhibits or reduces the production and/or deposition of amyloid-beta.

5. The use according to claim 1, wherein the medicament inhibits or reduces neurofibrillary tangles in the subject.

6. The use according to claim 1, wherein the medicament treats or reduces cognitive dysfunction in the individual.

7. Use according to claim 6, wherein the cognitive dysfunction is selected from the group consisting of learning disorders and memory disorders, disorientation, reduced cognitive function and dementia.

8. Use according to any one of claims 1 to 7, wherein the medicament comprises as the only active ingredient said koumine or a pharmaceutically acceptable salt thereof, or a combination of said koumine or a pharmaceutically acceptable salt thereof and one or more other active ingredients, and optionally a pharmaceutically acceptable carrier.

9. The use according to claim 8, wherein the pharmaceutically acceptable salt is selected from the group consisting of hydrochloride, sulfate, hydrobromide, hydroiodide, nitrate, bisulfate, phosphate, acid phosphate, citrate, acetate, oxalate, isonicotinate, lactate, salicylate, acid citrate, tartrate, oleate, tannate, pantothenate, bitartrate, ascorbate, succinate, maleate, gentisate, fumarate, gluconate, glucuronate, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate and pamoate.

10. The use according to claim 9, wherein the pharmaceutically acceptable salt is the hydrochloride salt.

11. Use according to claim 8, wherein the other active ingredients are selected from: a cholinesterase inhibitor; a statin-based drug; a beta secretase inhibitor; gamma secretase inhibitors; an a β aggregation inhibitor; a β -peptide specific antibody; a selective estrogen receptor molecule; an antihypertensive agent; an anti-inflammatory agent; an antioxidant; and combinations thereof.

12. Use according to claim 11, wherein the other active ingredients are selected from: alpha-blockers, angiotensin converting enzyme inhibitors, angiotensin receptor blockers, calcium channel blockers, diuretics, steroidal anti-inflammatory agents, and non-steroidal anti-inflammatory agents.

13. Use according to claim 11, wherein the other active ingredients are selected from:

tacrine hydrochloride, donepezil, rivastigmine and galantamine; prednisone, acetylsalicylic acid, diflunisal, salsalate, ibuprofen, dexibuprofen, naproxen, fenoprofen, dexketoprofen, flurbiprofen, oxaprozin, loxoprofen, indomethacin, tolmetin, sulindac, etodolac, ketorolac, diclofenac, nabumetone, piroxicam, meloxicam, tenoxicam, lornoxicam, isoxicam, mefenamic acid, meclofenamic acid, flufenamic acid, celecoxib, hyperforin; atorvastatin, rosuvastatin, fluvastatin, mevastatin, pitavastatin, pravastatin, rosuvastatin and simvastatin; CTS-21166, GRL-8234, TAK-070, and rosiglitazone; NIC 5-15; a metal ion chelating agent; palivizumab, sorafenib; raloxifene; valsartan; garlic extract, curcumin, melatonin, resveratrol, ginkgo biloba extract, green tea extract, vitamin C, vitamin E, selegiline; and any combination thereof.

14. Use according to claim 13, wherein the metal ion chelating agent is Cu²⁺、Zn²⁺Or Fe³⁺A chelating agent.

15. Use according to claim 13, wherein the metal ion chelating agent is clioquinol or PBT 2.

16. Use according to any one of claims 1 to 7, wherein the individual is a mammal.

17. Use according to any one of claims 1 to 7, wherein the individual is a human.

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