WO2006114019A1 - Extrait de polysaccharide a partir de lycium barbarum comme agent neuroprotecteur contre la neurotoxicite du peptide beta-amyloide - Google Patents

Extrait de polysaccharide a partir de lycium barbarum comme agent neuroprotecteur contre la neurotoxicite du peptide beta-amyloide Download PDF

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WO2006114019A1
WO2006114019A1 PCT/CN2005/000576 CN2005000576W WO2006114019A1 WO 2006114019 A1 WO2006114019 A1 WO 2006114019A1 CN 2005000576 W CN2005000576 W CN 2005000576W WO 2006114019 A1 WO2006114019 A1 WO 2006114019A1
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peptide
extract
lba
neurons
lbb
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PCT/CN2005/000576
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WO2006114019A8 (fr
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Chuenchung Raymond Chang
Waihung Yuen
Kwokfai So
S. Y. Zee
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Versitech Limited
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Publication of WO2006114019A8 publication Critical patent/WO2006114019A8/fr

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/81—Solanaceae (Potato family), e.g. tobacco, nightshade, tomato, belladonna, capsicum or jimsonweed
    • A61K36/815—Lycium (desert-thorn)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/18—Antipsychotics, i.e. neuroleptics; Drugs for mania or schizophrenia
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/24—Antidepressants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis

Definitions

  • the present invention relates to novel compositions from the extracts of the Lycium barbarum as a neuroprotective agent against ⁇ -amyloid peptide neurotoxicity which thus permits their use for the treatment of Alzheimer's disease (AD) and for the prevention of neuronal loss in aging against the accumulation of ⁇ -amyloid peptide in the brain.
  • the present invention relates to a method for obtaining compositions having anti- ⁇ -amyloid peptide neurotoxicity from Lycium barbarum, and formulations containing said compositions.
  • the present invention relates to methods of using stress kinases (c-Jun N-terminal kinase (JNK) and double-stranded RNA-dependent protein kinase (PKR)) as a technological platform for screening neuroprotective drugs in all kinds of natural and synthetic chemicals or formulations against ⁇ -amyloid peptides neurotoxicity.
  • stress kinases c-Jun N-terminal kinase (JNK) and double-stranded RNA-dependent protein kinase (PKR)
  • Alzheimer's disease is an age-related chronic neurodegenerative disease.
  • the major symptoms of Alzheimer's disease are cognitive and language impairment (Yankner, 1996; Ray et al., 1998; Bossy- Wetzel et al., 2004).
  • As it is a neurodegenerative disease all coordination of patient movement becomes a problem. It has been predicted that over 14 million US residents will suffer from this devastating disease (according to American Health Assistant Foundation). As aging populations rapidly increase in the next few decades, the occurrence of this disease will certainly be a major health problem and a burden on all governments in the world.
  • a major problem associated with Alzheimer's disease is neuronal loss.
  • the death processes for neurons can be mediated via neuronal apoptosis, granulovacuolar degeneration or synaptosis (Stadelmann et al., 1999; Engidawork et al., 2001; Su et al., 2001; Chang et al., 2002a; Leroy et al., 2002; Scheff and Price, 2003; Yu et al., 2004).
  • Neuronal apoptosis is mediated by different highly regulated biological processes. Activation of pro-apoptotic mechanisms have been found in the above three kinds of death processes.
  • caspases-3 and -8 have been found in postmortem human AD brain (Rohn et al., 2001; Su et al., 2001; Scheff and Price, 2003).
  • Our laboratory at the University of Hong Kong and other laboratories have also found activation of caspase-3 and a pro-apoptotic stress kinase, the double- stranded RNA-dependent protein kinase (PKR), in granulovacuolar degenerative neurons in postmortem human AD brain sections (Chang et al., 2002a). Therefore, combating the death processes for neurons against aging, biological stress, or environmental toxins is major task in developing treatments of Alzheimer's disease.
  • PLR double- stranded RNA-dependent protein kinase
  • Alzheimer's disease The pathological features of Alzheimer's disease are the appearance of senile plaques and neurofibillary tangles. Accumulation or so-called deposit of insoluble ⁇ - amyloid (A ⁇ ) peptide is the major component in such senile plaques (Ray et al., 1998). Therefore, it has been generally considered that increased production of insoluble A ⁇ peptide contribute to the formation of senile plaques. Indeed, it has been reported that production of insoluble A ⁇ peptide increases with age. High levels of A ⁇ peptide are toxic to neurons leading to neuronal apoptosis (Iversen et al., 1995; Saido, 2003; Bossy-Wetzel et al., 2004).
  • a ⁇ production in mutated human amyloid precursor protein (APP, K670N/M671L and V717F mutation) transgenic mice, and injection of A ⁇ into aged rhesus monkey or primate also show Alzheimer-type neuropathology, neuronal death and is correlated to memory deficits.
  • a ⁇ peptide has also been shown to exert neurotoxic effects leading to neuronal apoptosis in vitro in several laboratories. Therefore, A ⁇ peptide has long been regarded as a major toxin in the pathogenesis of Alzheimer's disease.
  • tau phosphorylation can also occur in other types of neurodegenerative diseases without the problem of dementia.
  • a ⁇ peptide neurotoxicity can trigger hyper-phosphorylation of tau. Therefore, it is expected that high levels of A ⁇ peptides would lead to cascades of pro-apoptotic pathways including the abnormal phosphorylation of tau protein.
  • Alzheimer's disease Since the major symptom in Alzheimer's disease is cognitive impairment, nearly all, if not 100%, of the current treatments for Alzheimer's disease are based on targeting anti-cholinesterase. Up to third generation anti-cholinesterase drugs have been employed in the clinic. However, most of these drugs have side effects that may be intolerable for the patients. In addition, these drugs slow down the cognitive impairment but are not intended to safeguard neurons. If neurons are undergoing degeneration, problem of cognitive impairment will still occur. Furthermore, in view of the increasing population of aged people in the world, it is better to protect neurons far before their commitment into the initiation of apoptosis. Therefore, neuroprotection will increasingly become a useful and effective therapeutic strategy for the prevention of neuronal death in Alzheimer's disease and aging.
  • JNK c-Jun N-terminal kinase
  • PSR double-stranded RNA-dependent protein kinase
  • polysaccharides extracts of Lycium barbarum has been shown to significantly prolong the half-death time for male Drosophila melanogaster (Xu et al., 2001). In addition, it can markedly attenuate hydroxy 1 radicals-induced lipid peroxidation in the liver of aged mice. Furthermore, it has been shown that polysaccharides extracts from Lycium barbarum can significantly reduce CCl 4 - triggered liver toxicity (Kim et al., 1999). Also, it can exert protective effects against hyperthermia- induced damage in cultured seminoferous epithelium (Wang et al., 2002). Based on all of this evidence taken together, we have now recognized that polysaccharides extracts from Lycium barbarum exert protective effects to neurons against environmental toxins like ⁇ -amyloid (A ⁇ ) peptides in AD.
  • a ⁇ ⁇ -amyloid
  • the present inventors have achieved this invention by proving that the said compositions from aqueous extracts of the fruit of Lyciurn barbarum provide neuroprotective effects to A ⁇ peptide toxicity. Also, the technological platform of using phosphorylation of JNK and PKR will be an effective method for screening neuroprotective drugs from all kinds of natural or synthetic chemicals or formulations against ⁇ -amyloid peptides neurotoxicity.
  • compositions having neuroprotection from Lycium barbarum are effective agents for protecting neuronal cell death induced by A ⁇ -peptide.
  • methods for obtaining compositions having neuroprotection from Lycium barbarum in addition, there are also provided formulations containing said composition.
  • methods of purification of the invention composition from Lycium barbarum, and the characterization thereof we have discovered.
  • the present invention provides novel therapeutic agents from the polysaccharides extracts from Lycium barbarum to prevent neuronal death and neuronal apoptosis in A ⁇ -peptide neurotoxicity and Alzheimer's disease. Also, the present invention provides a technological platform by using Western-blot analysis of PKR and JNK for drug screening to develop neuroprotective agent against neuronal loss in Alzheimer's disease.
  • LDH lactate dehydrogenase
  • Figure 1 presents the elution profile of neuroprotective preparation from the aqueous extract o ⁇ Lycium barbarum.
  • Figure 2 presents the elution profile of neuroprotective preparation from the basic aqueous extract oiLycium barbarum.
  • Figures 3 A to 3D collectively present the GC profile of neuroprotective preparation from the aqueous extracts o ⁇ Lycium barbarum.
  • Figure 3 A presents the GC profile of an aqueous extract (LBA; see Example 1);
  • Figure 3B presents the GC profile of LBA-AO,
  • Figure 3C presents the GC profile of LBA-Al and
  • Figure 3D presents the GC profile of LBA-A2 (see Example 2).
  • Figures 4A to 4D collectively present the GC analysis of neuroprotective preparation from the basic aqueous extracts of Lycium barbarum.
  • Figure 4A presents the GC profile of an basic aqueous extract (LBB; see Example 1);
  • Figure 4B presents the GC profile of LBB-BO,
  • Figure 4C presents the GC profile of LBB-Bl and
  • Figure 4D presents the GC profile of LBB-B2 (see Example 2).
  • Figure 5 shows the cytotoxicity analysis by the LDH activity assay.
  • Figure 6 shows activity of caspase-3.
  • Figure 7 shows the activity of caspase-2.
  • Figure 8 shows the cytotoxicity analysis by the LDH activity assay in neuronal cell cultured after treatment with A ⁇ -peptide or LBB (0.1 - 500 ⁇ g/ml).
  • Figure 9 shows caspase-3 activity after treatment with A ⁇ -peptide or LBB.
  • Figure 10 shows the activity of caspase-2.
  • Figure 11 shows the comparison of neuroprotective effects between LBA and LiCl.
  • Figure 12 shows the comparison of neuroprotective effects of LBA, LBB and heparin on neuronal morphology.
  • Figure 13 shows the Western-blot analysis of JNK and c-Jun in neurons treated with LBA and LBB.
  • Figure 14 shows the comparison of neuroprotective effects between the specific JNK inhibitor SP 600125 and LBA
  • Figure 15 shows the Western-blot analysis of PKR and eIF2 ⁇ in the neuroprotective effects of LBA and LBB.
  • Figure 16 shows the effects of different fractionated products from LBA on A ⁇ -peptide neurotoxicity by examining caspase-3 activity.
  • Figure 17 shows the neuroprotective effects of LBA- A2 against A ⁇ -peptide neurotoxicity in term of neuronal morphology.
  • Figure 18 shows the Western-blot analysis of LBA-A2.
  • Figure 19 shows the neuroprotective effects of fractionated products (LBB-Bl) from LBB on A ⁇ -peptide neurotoxicity by examining the caspase-3 activity.
  • Figure 20 shows the neuroprotective effects of fractionated products (LBB-B2) from LBB on A ⁇ -peptide neurotoxicity by examining the caspase-3 activity.
  • compositions in substantially purified form comprising:
  • water soluble polyanionic polysaccharide-containing extracts comprising arabinose, galactose, glucose, xylose, rhamnose, mannose, glucuronic and glactouronic acid as analyzed by gas chromatography;
  • compositions can be further characterized as:
  • novel compositions of the present invention can be prepared by a variety of methods for both extraction and purification. Such methods include the one described in the Example of the present specification. Briefly, the dry fruit is allowed to soak in ethanol or another lower alcohol which is then driven off by heating taking substantially all of the alcohol soluble components with it. The residue crushed into a powder and extracted with water resulting in an aqueous solution (which can be freeze-dried if desired) and a second residue.
  • One extract of the invention (LBA) can be realized by adding ethanol to the aqueous solution to form a precipitate, which is optionally further extracted with butanol and/or methanol.
  • a second extract of the invention can be realized by combining the second residue with an aqueous base, such as, for instance, sodium hydroxide, resulting in a basic aqueous solution which can be dialysized and concentrated, followed by adding ethanol to the aqueous solution to form a precipitate, which is optionally further extracted with methanol.
  • an aqueous base such as, for instance, sodium hydroxide
  • Both extracts can be fractionated by column chromatography using gradient aqueous base as an eluent.
  • the composition can be obtained from cells of the dry Lycium barbarum fruit by purifying the invention composition by contacting an extract from Lycium barbarum with an anion exchange material which selectively binds negatively charged materials, and recovering the invention composition from the anion exchange material.
  • the present invention is also directed to pharmaceutical formulations suitable for
  • any compatible carrier can be employed which is suitable for the type of dosage form employed.
  • Such formulations include solid dosage forms such as tablets, capsules, cachets, pellets, pills, powders or granules, topical dosage forms such as solutions, powders, fluid emulsion, fluid suspensions, semi-solids, ointments, pastes, reams, gels or jellies and foam, and parenteral dosage forms which include solutions, suspensions, emulsions or dry powers.
  • the means and methods for administration are known in the art and those skilled can refer to various pharmacological references for guidance. Some examples are “Modern Pharmaceutics", Banker & Rhodes, Marcel Dekker, Inc. 1979 and “Goodman & Gilman's The Pharmaceutical Basis of Therapeutics", MacMillan Publishing Co.. Appropriate dosage amounts can be readily determined by those of skill in this art.
  • composition of the present invention is preferably present in a purified form when administered.
  • invention compositions are obtained by extraction from Lycium barbarum, it is desirable to separate soluble extract from (residual) particulate matter by appropriate means (e.g., filtration, centrifugation, or other suitable separation techniques). Separation of solid materials may be performed one or more times during the extracting process.
  • suitable means e.g., filtration, centrifugation, or other suitable separation techniques. Separation of solid materials may be performed one or more times during the extracting process.
  • the utility of invention compositions as a therapeutic agent is enhanced by greater purification. Greater doses may be necessary when less pure forms of the extract are employed.
  • compositions are preferably substantially free from heavy metals, contaminating plant materials, contaminating microorganisms, oxalic acid or precursors of oxalic acid or any other contaminants which may be present in a preparation which can be derived from plant material.
  • invention compositions from the fruit of Lycium barbarum is the presently most practical method for obtaining such materials
  • the present invention also contemplates obtaining such materials from other sources such as other plants which may contain recoverable amount of compositions having the properties described herein.
  • Other plants contemplated include species within the family of Solanaceae, of which Lycium is a member.
  • invention compositions could be obtained by culturing plant cells, such as Lycium barbarum cells, in vitro and extracting the active ingredients from the cells or recovering the active ingredients from the cell culture medium.
  • extract means the active ingredients isolated from the fruit or other parts of Lycium barbarum or other natural sources including but not limited to all varieties, species, hybrids or genera of the plant regardless of the exact structure of the active ingredients, from or method of preparation or method of isolation.
  • extract is also intended to encompass salts, complexes and/or derivatives of the extract which possess the above-described biological characteristics or therapeutic indication.
  • extract is also intended to cover synthetically or biologically produced analogs and homologs with the same or similar characteristics yielding the same or similar biological effects of the present invention.
  • the purified composition contemplated for use herein include purified extract fractions having the properties described herein from any plant or species, preferably Lycium barbarum, in natural or in variant form, and from any source, whether natural, synthetic, or recombinant. Also, included within the scope of the present invention are analogs and homologs of the above-described purified compositions.
  • the present invention also contemplates the use of synthetic preparations having the characteristics of invention compositions. Such synthetic preparations could be prepared based on the chemical structure and/or functional properties of the above- described compositions of the present invention. Also contemplated are analogs and homologs of the chemical structure of the invention compositions and having the functional properties of compositions according to the present invention.
  • analogs and homologs of the invention compositions embraces compounds which differ from the structure of invention compositions by as little as the addition and/or replacement and/or deletion of one or more residues thereof. Such compounds in all instances, however, have substantially the same activity as invention compositions.
  • analogs refers to compounds having the same basic structure as invention compositions, but differ in one or more residues;
  • homologs refers to compounds which differ from invention compositions by the addition and/or deletion and/or replacement of a limited number of residues.
  • the extract was decanted to a clean container and the Lycium barbarum was extracted two more times with distilled water under the same conditions.
  • the extracts were poured through a cotton cloth to remove insoluble materials (residue).
  • the volume of the clarified extracts was reduced to about 20 litres by a rotary evaporator.
  • the condensed extract was freeze-dried. A total of 1800 g of dark brown dried powder was obtained.
  • the neuroprotective component in the resulting aqueous extract was precipitated by ethanol. To achieve this, 1800 g of the freeze-dried aqueous extract was dissolved in 20 L of water, and ethanol was added to a final concentration of 90% (vol/vol).
  • the residue was further incubated with a 5% sodium hydroxide solution (15 L) for 24 h.
  • the resulting basic extracts were poured through a cotton cloth to remove insoluble materials.
  • the volume of the basic extracts was exhaustively dialyzed (MW cutoff 3000 - 5000 Da) with running water for 60 h.
  • the volume of the dialyzate was reduced to about 8 L by a rotary evaporator.
  • the neuroprotective component in the resulting basic aqueous extract was precipitated by ethanol.
  • ethanol was added to the dialyzate to a final concentration of 90% (vol/vol).
  • the mixture was left for 24h and the precipitate was obtained by filtration through cotton and washed with 2 x 1.5 L of methanol. This yielded 75 g of dark brown powder, hereinafter designated LBB.
  • the neuroprotective component of the aqueous extract was further purified by gel filtration column chromatography.
  • An aqueous solution of LBA (2 g in 3 ml) was applied to a DEAE Sepharose Fast Flow column (100 x 2.6 cm) and eluted first with distilled water followed by 0.2M and 0.4M aqueous sodium hydroxide solution (gradient elution). Fractions of 10 ml were collected and the amount of carbohydrate in the collected fractions was monitor by phenol-sulfuric acid assay using water as the standard (see Figure 1).
  • the neuroprotective component of the basic aqueous extract was further purified by a similar method.
  • an aqueous solution of LBB (2 g in 5 ml) was applied to a DEAE Sepharose Fast Flow column (100 x 2.6 cm) and eluted first with distilled water followed by 0.2M and 0.4M aqueous sodium hydroxide solution (gradient elution) to give three fractions, hereinafter designated LBB-BO, LBB-Bl and LBB-B2 (see Fig. 2).
  • LBB-BO aqueous sodium hydroxide solution
  • the chemical nature of the aqueous extract and basic aqueous extract neuroprotective fractions was investigated by different chemical tests.
  • the glycosyl compositions analysis was performed by combined gas chromatography/mass spectrometry (GC/MS) of the per-O-trimethylsilyl (TMS) derivatives of the monosaccharide methyl glycosides produced from the sample by acidic methanolysis. To achieve this, methyl glycosides were first prepared from a portion of each dry
  • Elemental, infrared, NMR and other spectroscopic analytical means can also be employed to characterize the active compound.
  • FIG. 5 shows the percentage of total LDH release in the various treatment
  • Caspase-3 has long been regarded as an index for neuronal apoptosis. Therefore, the colorimetric caspase-3 activity assay was performed at the concentrations stated in FIG. 5, and neuronal apoptosis was determined by the colorimetric caspase-3 activity assay by measuring the absorbance (at 405 nm) of the yellow product (pNA) cleaved from the substrate. Results (***p ⁇ 0.001 vs. the group
  • ⁇ SE mean ⁇ SE from at least three independent experiments. As shown in FIG. 6, neurons exposed to A ⁇ -peptide displaced a two-fold increase in caspase-3 activity. However, neurons exposed to different concentrations of LBA (0.1 - 500 ⁇ g/ml)
  • ⁇ g/ml showed the best neuroprotective effect since it could reduce caspase-3 activity from 2.13 fold to 1.14 fold when compared with the A ⁇ -peptide treated group. Neurons exposed to the aqueous extract LBA per se did not induce a significant increase in caspase-3 activity.
  • Caspase-2 is an important index for neuronal apoptosis for ⁇ -amyloid peptide neurotoxicity. It has been reported that A ⁇ -peptide triggered activation of caspase-2. Also, it has been suggested that the roles of caspase-2 in A ⁇ -peptide neurotoxicity is more significant than that of caspase-3. Therefore, we used the colorimetric method to determine caspase-2 activity. For caspase-2 activity, the trend was similar to that of caspase-3. Neurons exposed to A ⁇ -peptide triggered a 1.6-fold increase in caspase-
  • FIG. 7 *p ⁇ 0.05 vs. the group treated with A ⁇ -peptide alone, by one-way ANOVA, Student-Newman-Keuls method).
  • LBB lactate dehydrogenase
  • FIG. 8 shows the percentage of total LDH release of various treatment groups. Neurons pre-treated with LBB of different concentrations (0.1 - 500 ⁇ g/ml) significantly
  • Caspase-3 is a general apoptosis marker.
  • the colorimetric caspase-3 activity assay (at 405 nm) was performed. LBB at low
  • LBB elicits biphasic neuroprotective effects to attenuate activation of caspase-3.
  • LBB was able to significantly attenuated A ⁇ -peptide-induced caspase-2 activity to about 1.0 to 1.3-fold of control (FIG. 10, *p ⁇ 0.05 vs. the group treated
  • LBB from 1 to 500 ⁇ g/ml per se did not trigger a significant increase in caspase-2 activity.
  • LiCl has been shown to protect neurons against A ⁇ -peptide toxicity. It is also a well-known Western medicine for the treatment of manic-depressive bi-polar disorder.
  • FIG. 11 shows the caspase-3 activity (in terms of
  • percent toxicity calculated as (s.a. ofAP ./ LBA -/sp- treated - s.a. control ) ⁇ (s.a. ⁇ - s.a. control ); expressed as mean +/- SE from at least 3 independent experiments) after LBA and
  • LiCl treatment The unit of concentration of LiCl was converted from mM to ⁇ g/ml for the purpose of comparison with LBA. LiCl was toxic to neurons at about 85 ⁇ g/ml
  • LiCl could effectively reduce A ⁇ -peptide-stimulated caspase-3 activity
  • Heparin is a well-defined glycoprotein that has been demonstrated to attenuate
  • a ⁇ -peptide neurotoxicity Since LBA and LBB contain both carbohydrate and amino acid, we use heparin to make a comparison of their protective effects. Neurons were pre-incubated with either heparin (1 ⁇ M), LBA (100 ⁇ g/ml) or LBB (500 ⁇ g/ml) for 1
  • H were neurons pre-incubated with heparin, and then without and with A ⁇ -peptide
  • JNK is one of the stress kinases involved in the processes of neuronal apoptosis in many types of stress responses. It has also been reported that JNK is
  • JNK Upon A ⁇ -peptide stimulation, JNK is activated by phosphorylation. Activated JNK further phosphorylates its substrate, c-Jun, which will in turn target on downstream effectors, causing apoptosis.
  • Neurons were pretreated with LBA (100 ⁇ g/ml), LBB (10 ⁇ g/ml and 500 ⁇ g/ml) or LiCl (2 mM and 8 mM) for 1 hour,
  • Neurons were pretreated with either SP600125 (5 ⁇ M to 20 ⁇ M) or
  • FIG. 14 shows the caspase-3 activity (in terms of percentage toxicity) after SP600125 and LBA treatment. The unit of concentration of SP600125 was converted
  • SP600125 was used at 4.4 ⁇ g/ml ' (20 ⁇ M). The toxicity was 100% when using
  • SP600125 at more than 20 ⁇ M (data not shown). SP600125 was effective from 1.1
  • Active fraction from the LBA extracts of Lycium barbarum reduce the activation of stress kinase PKR pathway in neurons triggered by ⁇ -amyloid peptide
  • Neurons were pre-treated with LBA-A2 at 10, 100 or 500 ⁇ g/ml for 1 h prior to the exposure to A ⁇ -peptide (25 ⁇ M) for 4 h. Protein was extracted for Western-blot analysis of phosphorylated PKR and eEF2 ⁇ . The protein levels of total
  • caspase-3 serves as an index for neuronal apoptosis.
  • pretreatment of LBB -B 2 also exhibit significant neuroprotective effects against A ⁇ neurotoxicity (FIG. 20).
  • a hundred microgram per micro-liter exerted a nearly 100% neuroprotective effects (FIG. 20).
  • LBB-B2/?er se did not elicit any neurotoxicity.
  • Saido TC Overview-A ⁇ metabolism: from Alzheimer research to brain aging control.
  • Saido TC (ed) A ⁇ metabolism and Alzheimer's disease” ⁇ l-16, (2003).

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Abstract

La présente invention a trait à des extraits de Lycium barbarum comme agent neuroprotecteur contre la neurotoxicité du peptide β-amyloïde permettant ainsi leur utilisation pour le traitement de la maladie d'Alzheimer et pour la prévention de la perte neuronale en vieillissant contre l'accumulation du peptide β-amyloïde dans le cerveau. Des stress kinases (la c-Jun N-terminal kinase et la protéine kinase dépendante d'ARN bicaténaire) sont utilisées comme plate-forme technologique pour le criblage de médicaments neuroprotecteurs.
PCT/CN2005/000576 2005-04-26 2005-04-26 Extrait de polysaccharide a partir de lycium barbarum comme agent neuroprotecteur contre la neurotoxicite du peptide beta-amyloide WO2006114019A1 (fr)

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DE112005001269T DE112005001269T5 (de) 2005-04-26 2005-04-26 Polysaccharidextrakt aus Lycium barbarum als neuroprotektives Mittel gegen ß-Amyloidpeptidneurotoxizität
PCT/CN2005/000576 WO2006114019A1 (fr) 2005-04-26 2005-04-26 Extrait de polysaccharide a partir de lycium barbarum comme agent neuroprotecteur contre la neurotoxicite du peptide beta-amyloide

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Cited By (11)

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WO2010073104A3 (fr) * 2008-12-23 2010-08-26 Carmel - Haifa University Economic Corp Ltd. Amélioration de la fonction cognitive
CN103030708A (zh) * 2012-12-13 2013-04-10 大兴安岭林格贝有机食品有限责任公司 一种枸杞多糖的纯化新方法
CN103342755A (zh) * 2013-06-03 2013-10-09 湖北大学 枸杞多糖均一级份ⅳ及其制备方法和应用
JP2015500805A (ja) * 2011-11-29 2015-01-08 ベイラー カレッジ オブ メディスンＢａｙｌｏｒ Ｃｏｌｌｅｇｅ Ｏｆ Ｍｅｄｉｃｉｎｅ 認知強化方法
CN104311690A (zh) * 2014-11-11 2015-01-28 济南凯因生物科技有限公司 一种枸杞多糖的提取方法
WO2016160594A1 (fr) * 2015-03-27 2016-10-06 The Research Foundation For The State University Of New York Procédés et matériaux visant à réduire les niveaux de protéine bêta-amyloïde chez un mammifère
CN111419868A (zh) * 2019-01-09 2020-07-17 宁夏天仁枸杞生物科技股份有限公司 枸杞多糖在制备修复损伤神经药物或保健品中的应用
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CN112442104A (zh) * 2019-08-29 2021-03-05 中国科学院大连化学物理研究所 发酵法制备枸杞活性肽及方法和应用与抑制剂、功能食品
US11160825B2 (en) 2013-09-19 2021-11-02 Research Foundation Of The State University Of New York Methods and materials for treating diabetes or liver steatosis
WO2024022468A1 (fr) * 2022-07-29 2024-02-01 海南先声药业有限公司 Utilisation de rhamnose dans la préparation d'un médicament pour le traitement ou la prévention de maladies neurodégénératives, composition pharmaceutique et son utilisation

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WO2010073104A3 (fr) * 2008-12-23 2010-08-26 Carmel - Haifa University Economic Corp Ltd. Amélioration de la fonction cognitive
JP2015500805A (ja) * 2011-11-29 2015-01-08 ベイラー カレッジ オブ メディスンＢａｙｌｏｒ Ｃｏｌｌｅｇｅ Ｏｆ Ｍｅｄｉｃｉｎｅ 認知強化方法
CN103030708A (zh) * 2012-12-13 2013-04-10 大兴安岭林格贝有机食品有限责任公司 一种枸杞多糖的纯化新方法
CN103342755A (zh) * 2013-06-03 2013-10-09 湖北大学 枸杞多糖均一级份ⅳ及其制备方法和应用
US11160825B2 (en) 2013-09-19 2021-11-02 Research Foundation Of The State University Of New York Methods and materials for treating diabetes or liver steatosis
CN104311690A (zh) * 2014-11-11 2015-01-28 济南凯因生物科技有限公司 一种枸杞多糖的提取方法
US10792301B2 (en) 2015-02-13 2020-10-06 The University Of Toledo Therapeutic polysaccharide midi-GAGR and related materials and methods
WO2016160594A1 (fr) * 2015-03-27 2016-10-06 The Research Foundation For The State University Of New York Procédés et matériaux visant à réduire les niveaux de protéine bêta-amyloïde chez un mammifère
US10639322B2 (en) 2015-03-27 2020-05-05 The Research Foundation For The State University Of New York Methods and materials for reducing amyloid beta levels within a mammal
EP3273972A4 (fr) * 2015-03-27 2018-10-31 The Research Foundation for The State University of New York Procédés et matériaux visant à réduire les niveaux de protéine bêta-amyloïde chez un mammifère
US11253538B2 (en) 2015-03-27 2022-02-22 The Research Foundation For The State University Of New York Methods and materials for reducing amyloid beta levels within a mammal
CN111419868A (zh) * 2019-01-09 2020-07-17 宁夏天仁枸杞生物科技股份有限公司 枸杞多糖在制备修复损伤神经药物或保健品中的应用
CN112442104A (zh) * 2019-08-29 2021-03-05 中国科学院大连化学物理研究所 发酵法制备枸杞活性肽及方法和应用与抑制剂、功能食品
WO2024022468A1 (fr) * 2022-07-29 2024-02-01 海南先声药业有限公司 Utilisation de rhamnose dans la préparation d'un médicament pour le traitement ou la prévention de maladies neurodégénératives, composition pharmaceutique et son utilisation

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