DE102008003467A1 - Flavonoids as agents for the prophylaxis and treatment of neurodegenerative and other protein deficiency diseases - Google Patents

Abstract

The invention relates to the use of synthetically produced or derived from plants flavonoids, their derivatives or degradation products for the therapeutic and prophylactic treatment of these aggregation and misfolding diseases. This includes, in particular, preparations of skullcap (Scutellaria) and plants with similar ingredients (eg parsley, celery and chamomile and chrysanthemums) and the ingredients contained therein in pure form or in combinations (a, baicalin, etc.) as therapeutic and / or prophylactic active substances in prion infections and other neurodegenerative protein deficiency diseases (Alzheimer's disease, Parkinson's disease, Huntington's disease, senile amyloidosis, tauopathies, etc.) in humans and animals.

Description

1. Technical area

With The present invention provides a new way to Treatment of protein deficiency and protein aggregation diseases presented.

The The invention relates to the use of synthetically produced (including chemical modifications eg acetylation, hydroxylation, Halogenation, methylation, carbonylation, alkylation, glycosylation, Esterification, oxidation, hydrolysis, condensation, polymerization) or plant-derived flavonoids, their derivatives or degradation products for the therapeutic and prophylactic treatment of these aggregation and misfolding diseases. These include, in particular, preparations from Scullcap (Scutellaria) and plants with similar Ingredients (eg parsley, celery and chamomile and chrysanthemums) and the ingredients contained therein in pure form or in combinations (including the already known substances baicalein, Baicalin etc.) as therapeutically and / or prophylactically effective Substances in prion infections and other neurodegenerative Protein Deficiency Diseases (Alzheimer's Disease, Parkinson's Disease) Disease, Huntington's disease, senile amyloidosis, tauopathies, etc.) in humans and animals. So far, for these protein deficiency and aggregation diseases are not sufficiently effective causative agents Therapy.

2. Protein Misfolding and Protein aggregation diseases

2.1 forms of the disease

The proteins involved in protein aggregation and misfolding diseases are listed in Table 1. Table 1: Proteins involved in protein aggregation and misfolding diseases protein illness α-synuclein Parkinson's, Lewy body dementia, multisystem atrophy Amyloid-β peptide Alzheimer's disease gelsolin Familial amyloidosis amylin Diabetes (Type II) huntingtin Chorea huntington Ataxin-1 Spinocerebellar ataxia Superoxide dismutase 1 Amyotrophic sclerosis dew Tauopathies (Alzheimer's disease, frontotemporal dementia) transthyretin senile amyloidosis Prion protein Creutzfeldt-Jakob Disease, vCJD, GSS, FFI, Kuru Bovine Spongiform Encephalopathy, Scrapie Tranthyretin senile amyloidosis (ATTR amyloidosis)

The molecular basis of these diseases is the misfolding and aggregation of a specific protein (see Table 1). This misfolding can occur either spontaneously or by a mutation in this protein, be triggered by a cofactor, aging processes or infections. During the aggregation process, the misfolded protein forms oligomers that assemble into multimers and / or fibrils. The misfolding is the cause of the disease either by loss of the physiological activity of the protein or by direct or indirect toxic effects on the surrounding cells or the whole organism. The aggregates can be deposited as miniscule particles (which can not be visualized by light or electron microscopy) or as amyloids (see below) in body cells. Neurodegenerative protein deficiency diseases primarily affect neurons, astrocytes, micro- and / or macroglial cells and affect the surrounding extracellular spaces or body fluids. After staining with Congo red they can be visualized. The aggregation is mostly with a refolding of the protein into β-sheet-rich structures connected. It is still unclear which component is responsible for the cytotoxic and neurotoxic effects during the aggregation process.

2.2 Mechanisms of inhibition

• a) Direkte Interaktion mit dem aggregierenden Protein
• b) Beeinflussung posttranslationaler Prozesse, die die Proteinaggregation fördern
• c) Auflösung der Fibrillen und Aggregate

The following different therapeutic strategies are currently being intensively pursued to treat these diseases:

• a) Direct interaction with the aggregating protein
• b) Influencing post-translational processes that promote protein aggregation
• c) Dissolution of fibrils and aggregates

a) Direct interaction with the aggregating protein

A Ability to inhibit aggregation is the use of small peptides that accumulate in the resulting fibrils and stop the aggregation process. These peptides were analyzed by means of "structure based designs "are synthesized and are effective in vitro Inhibitors of aggregation of the amyloid β-protein (Aβ) in Alzheimer's disease and of amylin in diabetes (Type II). The therapeutic applications are limited, however, since the peptides can be easily broken down in the body and immunogen are. Another danger is that during the inhibition process, can form toxic intermediates.

A Another possibility to find inhibitors is the testing substance libraries using high-throughput techniques (high-throughput screening). These screenings are using cell-based and cell-free Assays have been performed and have been able to handle numerous molecules identify the aggregation of amyloid β-protein, Inhibiting prions, tau protein and huntingtin in vitro. A therapeutic However, the effect of these proteins in vivo has not been described so far Service.

A third way to prevent aggregation is the stabilization of the native protein by direct binding of a molecule. This was already possible with Tansthyretin and Superoxide dismutase are shown.

Antiaggregatory agents include: N-methyl peptides ( Sciarretta et al 2006 ) β-sheet intercalators ( Sato-T et al. 2006 ) 4.5-dianilinophthalimides ( Blanchard-BJ et al. 2004 ) N744 ( Chirita et al. 2004 ) Juglone, Celastrol ( Wang et al ) C2-8 ( Zhang, et al. 2005 ) Fosfosal, levonordefrin, ( Desai et al. 2006 ) nadolol N'-benzylidene-benzohydrazide ( Bertsch et al., 2005 )

The substances that stabilize the native conformation include: Dicloflenac, diflunisal, ( Johnson et al. 2005 ) flufenamic

b) influencing post-translational processes, promoting protein aggregation

One Another starting point is the inhibition of secondary processes, that influence protein aggregation. An example is proteases, which proteolytically cleave the amyloid precursor protein (APP) and thereby responsible for Alzheimer's disease Form amyloid β-protein. By inhibition of these proteases (β- and γ-secretase) may therefore also accumulate of the Aβ protein. Various substances have already been found in vitro, specifically those proteases can inhibit. With a few exceptions, you can However, these substances do not cross the blood-brain barrier. Another problem is that by inhibiting especially the β-secretase toxic side effects occur, the other processes such as synaptic Affect plasticity and myelin formation of the nerves adversely can.

The phosphorylation of proteins is also a target in the search for effective substances. Examples are the (hyper) phosphorylation of the tau protein in tauopathies and the α-synuclein in Parkinson's disease. Starting point here is the inhibition of the corresponding kinases that ver for the phosphorylation ver are responsible. Also, various substances have been identified in vitro that inhibit the kinases. A serious problem, however, is that these kinases have multiple substrates and therefore toxic side effects occur again by inhibiting other metabolic pathways.

After all Oxidation processes are important in fibril formation Role. During the aggregation of Aβ, α-synuclein and huntingtin occur so-called "reactive oxygen species (ROS) "that are neurotoxic and above in addition, accelerate the aggregation itself. The usage of Antioxidants such as polyphenols (eg curcumin) and metal chelators is therefore a way to stop these processes or to stop. In vitro, inhibitory effects were already possible to be shown.

Substances that inhibit proteolysis of the amyloid precursor protein include: NSAID (inhibits γ-secretase) ( Beher et al. 2004 ). Lithium (inhibits γ-secretase) ( Phiel et al. 2003 ). Gleevec (inhibits γ-secretase / presilin) ( Netzer et al. 2003 ). MRK 560 (inhibits γ-secretase) ( Best et al. 2007 ) R-flurbiprofen (flurizan) (inhibits γ-secretase) ( Eriksen et al. 2003 ) Acylguanidines (inhibit β-secretase) ( Cole al. 2006 ). Carboyclic and heterocyclic ( Hanessian et al. 2005 ) Piptidomimetics (inhibit β-secretase)

Substances that inhibit phosphorylation include: AR-A014418 ( Bhat et al. 2003 ) lithium ( Noble et al. 2005 )

Substances that inhibit oxidation include: curcumin ( Yang et al. 2005 ) melatonin ( Matsubara et al. 2003 )

c) Resolution of the aggregates

A Another therapeutic option is the trial, the amplify protein degradation mechanisms present in the cell. For this one can either induce the formation of specific chaperones or to promote autophagic processes ("macroautophagy"). Chaperones serve for the correct folding of a protein or can misfolded proteins become cellular Perform removal. From a chaperone - hsp70- could be shown to both Aβ oligomers and mutated Apply huntingtin proteins to proteolytic degradation can. Various substances have already been identified which induce the formation of hsp70 and in vitro the formation of Aggregates inhibited. However, these substances are usually different Chaperone can inhibit serious side effects can not be excluded in vivo. Finally became in the aggregation of α-synuclein and mutant huntingtin observed that they move from the cell to certain regions, the so-called Aggresomes is transported. In these organelles are the Ag gregate either degraded by macroautophagy or innocuous Embedded fibrils. Therefore, a therapeutic target is the Increase in autophagic activity. There is a number of substances that have this effect are a problem However, there are proteins with anti-tumor properties be reduced.

Chaperone-enhancing substances include: geldanamycin ( Auluck et al. 2005 ) Celastrole ( Westerheide et al. 2004 )

Activation of the resolution of aggregates rapamycin ( Ravikumar et al. 2004 ) trehalose ( Sarkar et al., 2007 )

2.3 Inhibitors of prion diseases

Prion infections are transmissible protein deficiency diseases in humans and animals for which In-vi tro- as well as in-vivo (animal) models exist in which a possible prophylactic and / or therapeutic effect of chemical substances can be detected. Prion diseases (Creutzfeldt-Jakob disease, BSE, scrapie) are triggered by the misfolding of a body protein, the prion (PrP ^C ). This misfolding, also called conversion, can be triggered by infection, mutation, or sporadic (random) events. The misfolded protein (PrP ^Sc ) has the property of transferring this misfolding to other PrP ^C molecules. This causes aggregation and accumulation of misfolded PrP ^Sc molecules, especially in the CNS. Along with the formation of the aggregates, neurodegenerative events occur in the CNS that ultimately lead to death.

• • multicyclische anionische Moleküle (Congo Red): Gervasoni et al. 2004
• • polyanionische sulph. Polysaccharide (Pentosan Polysulphat): Ouidja et al. 2007
• • Polyene Antibiotika (Amphotericin B): McKenzie et al. 1994
• • Anthracycline (Iodoxorubicin): Tagliavini et al. 1997
• • cyclische Tetrapyrrole (Porphyrine, Phthalocyanine): Priola et al. 2000
• • Peptide (β-sheet breaker): Soto et al. 2000
• • *Acridine Derivate (Quinacrin): Benito-León et al. 2004
• • *Phenothiazin Derivate (Chlorpromazin): Korth et al. 2001
• • Polyamine (SuperFect; DOSPA): Supattapone et al. 2001
• • Induktoren der Aggregation (Suramin): Gilch et al. 2001
• • mAbs gegen PrP; rekombinante anti-PrP Fab-Fragmente: White et al. 2003
• • RNAi: Pfeifer et al. 2006
• • Curcumin: Caughey et al., 2003
• • Cyclodextrine: Prior et al. 2007

The following substances or substance classes have been described with which the prion infections and pathogenesis can be influenced:

• • multicyclic anionic molecules (Congo Red): Gervasoni et al. 2004
• • polyanionic sulph. Polysaccharides (pentosan polysulphate): Ouidja et al. 2007
• • Polyene antibiotics (Amphotericin B): McKenzie et al. 1994
• • Anthracycline (Iodoxorubicin): Tagliavini et al. 1997
• Cyclic tetrapyrroles (porphyrins, phthalocyanines): Priola et al. 2000
• Peptides (β-sheet breaker): Soto et al. 2000
• • * acridine derivatives (quinacrine): Benito-Leon et al. 2004
• • * Phenothiazine derivatives (chlorpromazine): Korth et al. 2001
• • Polyamines (SuperFect; DOSPA): Supattapone et al. 2001
• • Inducers of aggregation (suramin): Gilch et al. 2001
• MAbs to PrP; Recombinant anti-PrP Fab fragments: White et al. 2003
• • RNAi: Pfeifer et al. 2006
• Curcumin: Caughey et al., 2003
• Cyclodextrins: Prior et al. 2007

Alles However, these substances are characterized by a high toxicity and significant side effects, making a prophylactic or therapeutic dose in vivo is only marginal or absent. Furthermore, they often do not possess sufficient penetrance through the blood-brain barrier, leaving only an insufficient drug concentration in the CNS.

2.4 Conclusion

It It must be noted that there are different strategies for a prophylactic and / or therapeutic treatment for protein misfolding diseases gives. However, no chemical substance is available yet, sufficient for those on the living animal or in humans prophylactic or therapeutic effect could be shown. An important aspect of this is, above all, that in the majority the course of therapy has serious side effects the administered agent occurred.

3. Skullcap (scutellaria) and its ingredients

in the Within the scope of the present invention were two plant ingredients (Baicalein, Baicalin) from the group of flavonoids found that in vitro specifically the misfolding of the prion protein (conversion) inhibit and dissolve existing aggregates. These substances come from the scullcap Scutellaria lateriflora. Even the tea, which was prepared from this herb, has these in vitro activity. It was finally shown that the tea in the animal experiment the outbreak of the prion illness significantly can delay. Pathological investigations of the treated Mice showed that the application is harmless. There the molecular mechanisms of prion diseases are similar to those of other protein aggregation diseases are, can the effect of the ingredients from the Helmkraut is also transmitted to other diseases such as Alzheimer's, Parkinson's, etc. Helmkrauttee thus represents u. a. an effective therapeutic against these aggregation diseases Helmkraut, its ingredients and their previously known therapeutic Effectiveness received.

3.1. Analysis of scullcap (scutellaria) and its ingredients

The Helmet herbs (Scutellaria) belong to the family the Labiatae (Lamiaceae).

There are about 360 species known to be used in traditional medicine in Asia, North America and Europe ( Joshee et al. 2002 ). Tea preparations or extracts are said to be nonspecific anti-inflamma have toric, anti-viral, sedidative and neuroprotective effects. They are also said to have anti-cancerogenic effects and are used to treat epilepsy, insomnia and neuralgia. The effect is based on the ingredients of the helmet herb, the flavonoids.

3.2. flavonoids

The flavonoids are a group of water-soluble plant dyes and play an important role in the metabolism of many plants. They belong together with the phenolic acids to the polyphenols. So far, more than 8,000 polyphenols are known. Flavonoids can be divided into the following groups: flavonoids flavonols Morin, quercetin, rutin, kaempferol, myricetin, isorhamnetin flavones Baicalein, Luteolin, Apigenin, Morin, Acacetin, Diurinary Tangeretin. flavanols Catechin, gallocatechin, epicatechin, epigallocatechin gallate, theaflavin, thearubigin flavanones Hesperetin, naringenin, eriodictyol Flavanonole Taxifolin, epicatechin isoflavonoids Genistein, daidzein Antho cyanidine (Anthocyanins) Cyanidin, delphinidin, malvidin, pelargonidin, peonidin, chalcone isoflavones Biochanin A, equol

3.3. Previously known effects of flavonoids

• antibacterial: Cushnie TP, & Lamb AJ. (2005)
• Antiviral: Serkedjieva J et al. (2007)
• Anti-Inflammatory: Talhouk et al. (2007)
• Anti-carcinogen: Li al (2007)
• Anti-amyloid: Porat et al. (2006), Rivière et al. (2006), Shoval et al. (2007)

In however, these three publications have been invariably In vitro data on the antiamyloid effect of flavonoids are shown. So far, there are no data on the efficacy of flavonoids in vivo (eg in animal experiments).

3.4. Baicalein and Baicalin

Baicalein and Baicalin belong to the group of flavones. Baicalein is a 5,6,7-trihydroxy-flavone, Baicalin has an additional sugar ring compared to Baicalein and is called Biacalein-7-Glucuronide ( 8th ). Other flavones are luteolin (ingredient of parsley and celery), apigenin (ingredient of parsley, chamomile and celery), diosmetin, acetoacetin and tangeretin (ingredients of chrysanthemums).

3.5. Scutellaria species with baicalein and baicalin:

Both flavonoids can be isolated from a variety of Scutellaria species, with Scutellaria baicalensis being the most intensively studied species ( Kim et al. 2007 ). In addition, Baicalein and Baicalin were isolated from Scutellaria lateriflora ( Awadv et al. 2003 ). Other scutellaria species are: S. rivularis, S. discolour, S. indica, S. scadens ( Joshee et al., 2002 ).

3.6. Inhibitory effects of baicalein and Baicalin

3.6.1. Anti-aggregatory properties of Baicalein in vitro:

• a) Inhibition of aggregation of 1SS-alpha-lactalbumin ( Bomhoff et al. 2006 )
• b) Inhibition of aggregation of alpha-synuclein ( Zhu et al. 2004 )
• c) indirect inhibition of platelet aggregation by inhibition of oxygenases ( Kälvegren et al. 2007 )

Antiaggregatory Properties of Baicalein in vivo have not previously been described Service.

3.6.2. Antioxidant properties of Baicalein in vitro and in vivo:

• Ciesielska et al. 2002

3.6.3. Anti-cancerogenic effects of Baicalein in vitro

• Chao et al. (2007) ,

Object of the invention

Of the Invention was the object of finding substances for the therapeutic treatment of protein deficiency and aggregation diseases are suitable.

Solution of the task

The Task has been in accordance with the features of the claims solved. As a starting point and exemplary example were Prion diseases chosen.

With The present invention provides a new way to Treatment of protein misfolding and protein aggregation diseases presented.

The The invention relates to the use of synthetically produced (including chemically produced modifications thereof (eg Acetylation, hydroxylation, halogenation, methylation, carbonylation, alkylation, Glycosylation, esterification, oxidation, hydrolysis, condensation, Polymerization)) or plant-derived flavonoids whose Derivatives or degradation products for therapeutic and prophylactic Treatment of these protein deficiency and aggregation diseases. These include, in particular, preparations of scullcap (Scutellaria) and plants with similar ingredients (eg parsley, Celery and chamomile and chrysanthemums) and the contained therein Ingredients in their pure form or in combinations (including the already known substances Baicalein, Bai calin etc.). These substances are for prophylaxis and / or therapy in prion infections and other neurodegenerative protein deficiency and aggregation diseases (Alzheimer's disease, Parkinson's disease, Huntington's disease Disease, senile amyloidosis, tauopathies, etc.) in humans and Animal suitable.

The Invention also relates to preparations of skullcap (scutellaria) and related plants and ingredients contained therein in pure form or in combinations (including the already known substances Baicalein, Baicalin etc.), therefore, as a therapeutic agent and / or prophylactic in protein deficiency and aggregation diseases can be used by humans and animals.

The Invention is intended to be based on exemplary embodiments without reducing them to these examples.

embodiments

Evidence of the efficacy of flavonoids as inhibitors of protein deficiency and aggregagtion diseases

The Effectiveness of flavonoids against these diseases could by the example of the helmet and its ingredients in the frame from in vivo studies on living organism. In addition, the antiaggregatory activity could also in be detected in vitro. The substances not only inhibit the New formation, but also lead to the dissolution already existing aggregates.

example 1

Effect in vitro

The efficacy was tested in vitro in 2 test systems. In a cell-free conversion assay, recombinantly produced prion protein is incubated together with infectious PrP ^Sc . By means of specific antibodies, the subsequent misfolding (conversion) of the prion protein as well as the susceptibility of the PrP ^Sc aggregates to the proteolytic degradation can be detected. This assay was developed by the authors / inventors and has already been published ( Eiden et al. 2006 ). The second, cell-based assay uses cells that are permanently infected with scrapie pathogens. Here the PrP ^Sc formation can be detected by immunochemical methods.

Example 2

Cell-free assay

The cell-free assay showed that the flavonoids Baicalein as well as Baicalin can inhibit the conversion of the prion protein in a concentration-dependent manner ( 1 ). Bacalein has a much higher (IC ₅₀ <20 μM) activity than Baicalin. Other structurally similar flavonoids (epicatechin, morin, naringenin, quercitin, kaempferol) had no inhibitory activity in the studies so far performed. In this assay it could also be shown that both flavonoids also make the already existing PrP ^Sc aggregates (amyloid) susceptible to proteolytic degradation ( 2 ). This degradation can be done by cellular or exogenously supplied proteases. Again, Baicalein has a much higher activity than Baicalin. The other flavoids studied were again ineffective. The tea of the herb itself can both inhibit the conversion concentration-dependent and make the aggregates susceptible to proteolytic degradation ( 3 ). The results show that Baicalein / Baicain interact specifically with PrP ^C molecules to prevent conversion. In addition, because of their planar structure, you may also be able to store in PrP ^Sc aggregates and then make them proteolytically accessible.

Example 3

Cell-based assay

In the second, cell-based assay, the anti-prion effect could also be detected. This involved working with cells that are permanently infected with a scrapie pathogen. Again, both baicalein, baicalin ( 4 ) as well as the tea extract ( 5 ) inhibitory effect, which was concentration-dependent. Baicalein in turn has a much higher activity than Baicalin ( 6 ). In a cell cytotoxicity test (MTT test) both substances and the tea showed no cytotoxic effects. However, as the other flavonoids are also inhibitory, it can be assumed that in addition to the specific effects of baicalein / baicalin, the group of polyphenolic flavonoids has a general inhibitory effect on PrP ^Sc formation. This may be due to the antioxidant effects of this group of substances.

Example 4

Effect in vivo

Finally, the therapeutic effect of the skullcap was investigated in a mouse model. For this purpose, six wild-type mice were used, which were infected (inoculated) intracerebrally (ic) with a scrapie pathogen ( 7 ). Two weeks prior to inoculation, the infusion of the mice was switched from water to freshly brewed tea and maintained until death. As a control group 6 mice were used, which were also infected ic, but were further soaked in water. In addition, 3 mice were used, which were soaked with the Helmkraut tea at the same time, but were not infected. The untreated infected mice died within a narrow time window of 147.3 days, while the tea-treated animals lived significantly longer with one exception (148 days). Three mice lived 60 days longer than the animals in the untreated control group. The uninfected but tea-treated animals showed no pathological changes indicative of a side effect from the flavonoids.

It has been shown that tea (extract with hot water) of the skullcap (scutellaria) the In incubation time of a prion infection, ie by up to 60 days, which corresponds to a pathogen titer reduction by a factor of 100,000. The effect is probably based on 2 ingredients, the flavonoids Baicalein and Baicalin.

The Results show that the treatment of scrapie-infected animals with scotch tea has a significant therapeutic effect. This effect can be reduced to a specific Interaction of Baicalein / Baicalin with the misfolded or aggregated prion protein to another on general neuroprotective effects of flavonoids.

Legend to the figures:

1 : Inhibition of PrP ^res formation in cell-free conversion assay.

• A: Baicalein inhibiert effizient die Bildung von PrP^res. In den Spuren 1–2 sind Proteinase K (PK)-resistente PrP^res Fragmente in Proben ohne Baicalein zu sehen, in den Spuren 3–4 Pro ben nach Inkubation mit 10.0 mM Baicalein, in den Spuren 5–6 mit 1.0 mM Baicalein, and in den Spuren 7–8 nach Inkubation mit 0.1 mM Baicalein.
• B: Hemmung der PrP^res Bildung durch Baicalin. PK-resistente Fragmente ohne Baicalin (Spuren 1–2), mit 10 mM Baicalin (Spuren 3–4), 1.0 mM Baicalin (Spuren 5–6) und 0.1 mM Baicalin (Spuren 7–8).
• C: Keine Hemmung der PrP^res-Bildung durch das Flavonoid Epicatechin: Proben ohne Epicatechin (Spuren 1–2), nach Inkubation mit 10 mM Epicatechin (Spuren 3–4), mit 1.0 mM Epicatechin (Spuren 5–6) und 0.1 mM Epicatechin (Spuren 7–8).

The detection in Western blot was carried out with the monoclonal antibody P4 by means of chemiluminescence.

• A: Baicalein efficiently inhibits the formation of PrP ^res . In lanes 1-2 are proteinase K (PK) to see -resistant PrP ^res fragments in samples without baicalein, in lanes 3-4 Pro ben after incubation with 10.0 mM baicalein, in lanes 5-6 with 1.0 mM baicalein, and in lanes 7-8 after incubation with 0.1 mM Baicalein.
• B: inhibition of PrP ^res formation by Baicalin. PK-resistant fragments without baicalin (lanes 1-2), with 10 mM Baicalin (lanes 3-4), 1.0 mM Baicalin (lanes 5-6) and 0.1 mM Baicalin (lanes 7-8).
• C: No inhibition of PrP ^res formation by the flavonoid epicatechin: samples without epicatechin (lanes 1-2), after incubation with 10 mM epicatechin (lanes 3-4), with 1.0 mM epicatechin (lanes 5-6) and 0.1 mM Epicatechin (lanes 7-8).

2 : Dissolution of PrP ^Sc aggregates in the cell-free conversion assay.

• A: Baicalein löst PrP^Sc-Aggregate effizient nach Inkubation mit PK auf Spuren 1–2 zeigen PK-resistente PrP^Sc-Aggregate ohne Baicalein, Spuren 3–4 Proben nach Inkubation mit 10.0 mM Baicalein, Spuren 5–6 mit 1.0 mM Baicalein, und Spuren 7–8 Proben nach Inkubation von 0.1 mM Baicalein.
• B: Auflösung der PrP^Sc-Aggregate durch Baicalin. PK-resistente Aggregate ohne Baicalin (Spuren 1–2), nach Inkubation mit 10 mM baicalin (Spuren 3–4), 1.0 mM baicalin (Spuren 5–6) und 0.1 mM baicalin (Spuren 7–8).
• C: Keine Auflösung der PrP^Sc-Aggregate durch Epicatechin: Proben ohne Epicatechin (Spuren 1–2), nach Inkubation mit 10 mM Epicatechin (Spuren 3–4), mit 1.0 mM Epicatechin und 0.1 mM Epicatechin.

These are the same Westernblots of 1 , which after removal of the antibody by means of "stripping" with a new antibody (Ra 10) were re-incubated.

• A: Baicalein dissolves PrP ^Sc aggregates efficiently after incubation with PK on lanes 1-2 show PK-resistant PrP ^Sc aggregates without baicalein, lanes 3-4 samples after incubation with 10.0 mM Baicalein, lanes 5-6 with 1.0 mM Baicalein, and Lanes 7-8 samples after incubation of 0.1 mM Baicalein.
• B: dissolution of the PrP ^Sc aggregates by Baicalin. PK-resistant aggregates without baicalin (lanes 1-2), after incubation with 10 mM baicalin (lanes 3-4), 1.0 mM baicalin (lanes 5-6) and 0.1 mM baicalin (lanes 7-8).
• C: No dissolution of PrP ^Sc aggregates by epicatechin: samples without epicatechin (lanes 1-2), after incubation with 10 mM epicatechin (lanes 3-4), with 1.0 mM epicatechin and 0.1 mM epicatechin.

3 : Inhibition of PrP ^res formation and dissolution of PrPSc aggregates in a cell-free conversion assay by Helmkrauttee.

• A: Helmkrauttee hemmt die Bildung of PrP^res. Spuren 1–2 zeigen PK-resistente PrP^res-Fragmente in Proben ohne Tee, Spuren 3–4 Proben nach Inkubation mit Tee, Spuren 5–6 mit Tee (1:10 Verdünnung), und Spuren 7–8 nach Inkubation mit Tee (1:100 Verdünnung).
• B: Helmkrauttee löst PrP^Sc-Aggregate nach Inkubation mit PK auf.

The detection in Western blot was carried out with the monoclonal antibody P4 by means of chemiluminescence.

• A: Helmkrauttee inhibits the formation of PrP ^res . Lanes 1-2 show PK-resistant PrP ^res fragments in samples without tea, lanes 3-4 samples after incubation with tea, lanes 5-6 with tea (1:10 dilution), and lanes 7-8 after incubation with tea ( 1: 100 dilution).
• B: Helmkrauttee dissolves PrP ^Sc aggregates after incubation with PK.

These are the same Western blots of Figure that were "incubated" after removal of the antibody by stripping with a new antibody (Ra 10). Lanes 1-2 show PK-resistant PrP ^Sc aggregates without tea, lanes 3-4 samples after incubation with tea, lanes 5-6 with tea (1:10 dilution), and lanes 7-8 after incubation with tea (1: 100 Dilution).

4 , Inhibition of PrP ^Sc formation in ScN ₂ A and SMB cells in a cell-based assay

• A: Zellen wurden mit abnehmenden Konzentrationen an Baicalein behandelt: Inkubation von ScN₂A Zellen mitl.0 mM Baicalein (Spur 1), 0.1 mM Baicalein (Spur 2), 0.01 mM Baicalein (Spur 3) und unbehandelte ScN₂A-Zellen. (Spur 4). Inkubation von SMB Zellen mit 1.0 mM Baicalein (Spur 5), 0.1 mM Baicalein (Spur 6), 0.01 mM Baicalein (Spur 7) und unbehandelte SMB-Zellen (Spur 8).
• B: Zellen wurden mit abnehmenden Konzentrationen an Baicalin behandelt: Inkubation von ScN2A Zellen mitl.0 mM Baicalin (Spur 1), 0.1 mM Baicalin (Spur 2), 0.01 mM Baicalin (Spur 3) und unbehandelte ScN2A-Zellen. (Spur 4). Inkubation von SMB Zellen mit 1.0 mM (Spur 5), 0.1 mM Baicalin (Spur 6), 0.01 mM Baicalin (Spur 7) und unbehandelte SMB-Zellen (Spur 8).
• C: Zellen wurden mit abnehmenden Konzentrationen an Epicatechin behandelt: Inkubation von ScN₂A Zellen mitl.0 mM Epicatechin (Spur 1), 0.1 mM Epicatechin (Spur 2), 0.01 mM Epicatechin (Spur 3) und unbehandelte ScN₂A-Zellen. (Spur 4). Inkubation von SMB Zellen mit 1.0 mM (Spur 5), 0.1 mM Epicatechin (Spur 6), 0.01 mM Epicatechin (Spur 7) und unbehandelte SMB-Zellen (Spur 8).

PK-resistant PrP signals on sections of a 96-well dot blot after detection with the pAb Ra 10.

• A: Cells were treated with decreasing concentrations of Baicalein: Incubation of ScN ₂ A cells with 1.0 mM Baicalein (lane 1), 0.1 mM Baicalein (lane 2), 0.01 mM Baicalein (lane 3) and untreated ScN ₂ A cells. (Lane 4). Incubation of SMB cells with 1.0 mM Baicalein (lane 5), 0.1 mM Baicalein (lane 6), 0.01 mM Baicalein (lane 7) and untreated SMB cells (lane 8).
• B: Cells were treated with decreasing concentrations of Baicalin: Incubation of ScN2A cells with 1.0 mM Baicalin (lane 1), 0.1 mM Baicalin (lane 2), 0.01 mM Baicalin (lane 3) and untreated ScN2A cells. (Lane 4). Incubation of SMB cells with 1.0 mM (lane 5), 0.1 mM Baicalin (lane 6), 0.01 mM Baicalin (lane 7) and untreated SMB cells (lane 8).
• C: Cells were treated with decreasing concentrations of epicatechin: Incubation of ScN ₂ A cells with 1.0mM epicatechin (lane 1), 0.1mM epicatechin (lane 2), 0.01mM epicatechin (lane 3), and untreated ScN ₂ A cells. (Lane 4). Incubation of SMB cells with 1.0 mM (lane 5), 0.1 mM epicatechin (lane 6), 0.01 mM epicatechin (lane 7) and untreated SMB cells (lane 8).

• A: Helmkrauttee inhibiert die PrP/PrP^res-Bildung in ScN₂A Zellen. Spur 1 zeigt das PK-resistent Fragment der unbehandelten Kontrolle, die Spuren 2–5 Proben, die mit zunehmenden Konzentrationen an Helmkrauttee inkubiert wurden: 1:500 (Spur 2), 1: (Spur 3), 1:20 (Spur 4) and 1:10 (Spur 5).
• B: Helmkrauttee inhibiert die PrP/PrP^res-Bildung in SMB-Zellen. Spur 1 zeigt das PK-resistent Fragment der unbehandelten Kontrolle, die Spuren 2–5 Proben, die mit zunehmenden Konzentrationen an Helmkrauttee inkubiert wurden: 1:500 (Spur 2), 1:50 (Spur 3), 1:20 (Spur 4) and 1:10 (Spur 5).

5 , Inhibition of PrP / PrP ^res in scrapie-infected cells formation after incubation with Helmkrauttee

• A: Helmkrauttee inhibits the PrP / PrP ^res formation in ScN ₂ A cells. Lane 1 shows the PK-resistant fragment of the untreated control, the lanes 2-5 samples incubated with increasing concentrations of helmet-scab tea: 1: 500 (lane 2), 1: (lane 3), 1:20 (lane 4) and 1:10 (track 5).
• B: Helmkrauttee inhibits the PrP / PrP ^res formation in SMB cells. Lane 1 shows the PK-resistant fragment of the untreated control, the lanes 2-5 samples incubated with increasing concentrations of scullcap tea: 1: 500 (lane 2), 1:50 (lane 3), 1:20 (lane 4 ) and 1:10 (lane 5).

The Detection in Western blot was carried out with the polyclonal antibody Ra10 by means of chemiluminescence.

6 : EC50 values of the investigated flavonoids

7 : Survival times of the mice after infection with the mouse scrapie strain RML compared to control mice.

C57 / B16 mice were inoculated intracerebrally with 30 μl of a 1% RML brain homogenate.
Blue line: Treatment with helmet herb tea. The survival times were up to 207 days.
Red line: control mice. All control mice died between 145-153 days.

8th : Structure of Baicalein and Baicalin

Bibliography:

• Awad, R, Arnason, J., Trudeau, V, Bergeron, C., Budzinski, JW, Foster, BC, Merali. (2003) Phytochemical and biological analysis of skullcap (Scutellaria lateriflora L.): A medical plant with anxiolytic properties. Phytomedicine 10: 640-643 ,
• Auluck PK, Meulener MC, Bonini NM. (2005) Mechanisms of Suppression of {alpha} Synuclein Neurotoxicity by Geldanamycin in Drosophila. J Biol Chem. 280: 2873-8 ,
• Beher D, Clarke EE, Wrigley JD, Martin AC, Nadin A, Churcher I, Shearman MS. (2004) Selected non-steroidal anti-inflammatory drugs and their gamma-secretase at a novel site. Evidence for an allosteric mechanism. J Biol Chem. 279: 43419-26 ,
• Benito-León J. (2004) Combined quinacrine and chlorpromazine therapy in fatal familial insomnia. Clin Neuropharmacol. 27: 201-3 ,
• Bertsch U, Winklhofer KF, Hirschberger T, Bieschke J, Weber P, Hartl FU, Tavan P, Tatzelt J, Kretzschmar HA, Giese A. (2005) Systematic identification of antiprion drugs by high throughput screening based on scanning for intensely fluorescent targets. J Virol. 79: 7785-91 ,
• Best JD, Smith DW, Reilly MA, O'Donnell R, Lewis HD, Ellis S, Wilkie N, Rosahl TW, Laroque PA, Boussiquet-Leroux C, Churcher I, Atack JR, Harrison T, Shearman MS. (2007). The novel gamma secretase inhibitor N- [cis-4 - [(4-chlorophenyl) sulfonyl] -4- (2,5-difluorophenyl) cyclohexyl] -1,1,1-trifluoromethanesulfonamide (MRK-560) reduces amyloid plaque deposition without evidence of notch-related pathology in the Tg2576 mouse. J Pharmacol Exp Ther. 2007 Feb; 320 (2): 552-8 ,
• Bhat R, Xue Y, Mountain S, Hellberg S, Ormo M, Nilsson Y, Radeseer AC, Jerning E, Markgren PO, Borgegård T, Nylöf M, Giménez-Cassina A, Hernández F, Lucas JJ, Díaz-Nido J, Avila (2003) J. Structural insights and biological effects of glycogen synthase kinase 3-specific inhibitor AR-A014418. J Biol Chem. 278: 45937-45 ,
• Blanchard BJ, Chef A, Rozeboom LM, Stafford KA, Weigele P, Ingram VM. (2004) Efficient reversal of Alzheimer's disease fibril formation and elimination of neurotoxicity by a small molecule. Proc Natl Acad Sci USA. 101: 14326-32 ,
• Bomhoff, G., Sloan, K., McLain, C., Gogol, EP, Fisher, MT (2006). The effects of the flavonoid baicalein and osmolytes on the Mg 2+ accelerated aggregation / fibrillation of carboxymethylated bovine 1SS-alpha-lactalbumin. Arch Biochem Biophys. 453: 75-86 ,
• Caughey B, Raymond LD, Raymond GJ, Maxson L, Silveira J, Baron GS. (2003) Inhibition of protease-resistant prion protein accumulation in vitro by curcumin. J Virol. 2003 May; 77 (9): 5499-502 ,
• Caughey WS, Priola SA, Kocisko DA, Raymond LD, Ward A, Caughey B. (2007). Cyclic tetrapyrrole sulfonation, metals, and oligomerization in anti-prion activity. Antimicrob Agents Chemother. 51 (11): 3887-94
• Chao JI, Su WC, Liu HF. (2007) Baicalin induces cancer cell death and proliferation retardation by the inhibition of CDC2 kinase and survivin associated with opposing role of p38 mitogen-activated protein kinase and AKT. Mol Cancer Ther. 6: 3039-48 ,
• Chirita C, Necula M, Kuret J. (2004) Ligand-dependent inhibition and reversal of tau filament formation. Biochemistry. 43: 2879-87 ,
• Ciesielska E, Gwardy's A, Metodiewa D. (2002) Anticancer, antiradical and antioxidant actions of novel Antoksyd S and its major components, baicalin and baicalein. Anticancer Res. 22: 2885-91 ,
• Cole DC, Manas ES, Stock JR, Condon JS, Jennings LD, Aulabaugh A, Chopra R, Cowling R, Ellingboe JW, Fan KY, Harrison BL, Hu Y, Jacobsen S, Jin G, Lin L, Lovering FE, Malamas MS, Steel ML, J Beach, Sukhdeo MN, Svenson K, Turner MJ, Wagner E, Wu J, Zhou P, Bard J. (2006) Acylguanidines as small-molecule beta-secretase inhibitors. J Med Chem. 49: 6158-61
• Cushnie TP, Lamb AJ. (2005) Antimicrobial activity of flavonoids. Int J Antimicrob Agents. 26 (5): 343-56 ,
• Desai UA, Pallos J, Ma AA, Stockwell BR, Thompson LM, Marsh JL, Diamond MI. (2006) Biologically active molecules that reduce polyglutamine aggregation and toxicity. Hum Mol Genet. 15: 2114-4 ,
• Engelstein, R., Ovadia, H. and Gabizon, R. (2007) Copaxone interferes with the PrPSc-GAG interaction. EJ Neurol. 14, 877-884 ,
• Eiden M, Palm GJ, Hinrichs W, Matthey U, Tooth R, Groschup MH. (2006) Synergistic and strain-specific effects of bovine spongiform encephalopathy and scrapie prions in the cell-free conversion of recombinant prion protein. J Gen Virol. 87: 3753-61 ,
• Eriksen JL, Sagi SA, Smith TE, Weggen S, The P, McLendon DC, Ozols VV, Jessing KW, Zavitz KH, Koo EH, Golde TE. (2003) NSAIDs and enantiomers of flurbiprofen target gamma-secretase and lower Abeta 42 in vivo. J Clin Invest. 112: 440-9
• Forloni, G., Iussian, S., Awan, T., Colombo, L., Angeretti, N., Girala, L., Bertani, I., Poli, G., Caramelli, M., Buzzone, M. G., Farina, L., Limido, L., Rossi, G., Giaccone, G., Ironside, J. W., Bugiani, O., Salmona, M. and Tagliavini, F. (2002). tetracyclines affect prion infectivity PNAS. 99, 10849-10854
• Gervasoni M, Pirola R, Pollera C, Villa S, Cignarella G, Mantegazza P, Poli G, Bareggi SR. (2004) Pharmacokinetics and distribution of sodium 3,4-diaminonaphthalenes-1-sulfonates, a Congo Red derivative active in inhibiting PrP (res) replication. J Pharm Pharmacol. 56: 323-8 ,
• Gilch, S, Winklhofer, KF, Groschup, MH, Nunziante, M, Lucassen, R, Spielhaupter, C, Muranyi, W, Tatzelt, J, Schätzl, HM (2001). Intracellular re-routing of prion protein prevents propagation of PrPSc and delays onset of prion disease. EMBO J. 20: 3957-3966 ,
• Hanessian S, Yun H, Hou Y, Yang G, Bayrakdarian M, Therrien E, Moitessier N, Roggo S, Veenstra S, Tintelnot-Blomley M, Rondeau JM, Ostermeier C, Strauss A, Ramage P, Paganetti P, Neumann U, Betschart C. (2005) Structure-based design, synthesis, and memapsin 2 (BACE) inhibitory activity of carbocyclic and heterocyclic peptidomimetics. J Med Chem. 48: 5175-90 ,
• Johnson SM, Wiseman RL, Sekijima Y, Green NS, Adamski-Werner SL, Kelly JW. (2005) Native state kinetic stabilization as a strategy to ameliorate protein misfolding diseases: a focus on the transthyretin amyloidoses. Acc Chem. Res. 38: 911-21 ,
• Joshee N, Thomas SP, Rao SM, and Anand KY (2002). Skullcap: Potential medicinal crop Trends in new crops and new uses. 580-586 ,
• Kälvegren H, Andersson J, Grenegard M, Bengtsson T. (2007) Platelet activation triggered by Chlamydia pneumoniae is antagonized by 12-lipoxygenase inhibitors but not cyclooxygenase inhibitors. Eur J Pharmacol 566: 20-7 ,
• Kim YH, Jeong DW, Kim YC, Son DH, Park ES, Lee HS. (2007) Pharmacokinetics of baicalein, baicalin and wogonin after oral administration of a standardized extract of Scutellaria baicalensis, PF-2405 in rats. Arch Pharm Res. 30: 260-5
• Korth C, May BC, Cohen FE, Prusiner SB. (2001) Acridine and phenothiazine derivatives as pharmacotherapeutics for prion disease. Proc Natl Acad Sci USA98: 9836-41 ,
• Copper L, Eiden M, Bushman A, Groschup MH. (2007) Amino acid sequence and prion strain specific effects on the in vitro and in vivo convertibility of ovine / murine and bovine / murine prion protein chimeras. Biochim Biophys Acta. 1772: 704-13 ,
• Li Y, Fang H, Xu W. (2007) Recent advance in the research of flavonoids as anticancer agents. Mini Rev Med Chem. 7: 663-78 ,
• Mange, A., Nishida, N., Milhavet, Milhavet, O., McMahon, HE, Casanova, D., Lehmann S. (2000). Amphotericin B inhibits the generation of the scrapie isoform of the prion protein in infecting cultures. J Virol. 74: 3135-40 ,
• Matsubara E, Bryant-Thomas T, Pacheco Quinto J, Henry TL, Poeggeler B, Herbert D, Cruz-Sanchez F, Chyan YJ, Smith MA, Perry G, Shoji M, Abe K, Leone A, Grundke Ikbal I, Wilson GL, Ghiso J, Williams C, Refolo LM, Pappolla MA, Chain DG, Neria E. (2003) Melatonin increases survival and inhibits oxidative and amyloid pathology in a transgenic model of Alzheimer's disease. J Neurochem. 85 (5): 1101-8. Erratum in: J Neurochem. (2003) 86: 1312
• May, BCH, Zorn, JA, Witkop, J., Sherrill, J., Wallace, AC, Legname, G., Prusiner, SB, Cohen, FE (2007). Structure-activity relationship study of prion inhibition by 2-aminopyridine-3,5-dicarbonitrile-based compounds: parallel synthesis, bioactivity, and in vitro pharmacokinetics. J. Med. Chem. 50: 65-73 ,
• McKenzie D, Kaczkowski J, Marsh R, Aiken J. (1994) Amphotericin B delays both scrapie agent replication and PrP-res accumulation early in infection. J Virol. 68: 7534-6 ,
• Murakami-Kubo, I., Doh-Ura, K., Ishikawa, K., Kawatake, S., Sasaki, K., Kira, J., Ohta, S., Iwaki, T. (2004) Quinoline derivatives are therapeutic candidates for transmissible spongiform encephalopathies. J Virol. 78: 1281-8 ,
• Netzer WJ, Dou F, Cai D, Veach D, Jean S, Li Y, Bornmann WG, Clarkson B, Xu H, Greengard P. Gleevec inhibits beta-amyloid production but not notch cleavage. Proc Natl Acad Sci USA 100: 12444-9 ,
• Noble W, Planel E, Zehr C, Olm V, Meyerson J, Suleman F, Gaynor K, Wang L, LaFrancois J, Feinstein B, Burns M, Krishnamurthy P, Wen Y, Bhat R, Lewis J, Dickson D, Duff K (2005) Inhibition of glycogen synthase kinase-3 by lithium correlates with reduced tauopathy and degeneration in vivo. Proc Natl Acad Sci USA 102: 6990-5 ,
• Ouidja MO, Petit E, Kerros ME, Ikeda Y, Morin C, Carpentier G, Barritault D, Brugere-Picoux J, Deslys JP, Adjou K, Papy-Garcia D. (2007) Structure-activity studies of heparin mimetic polyanions for anti -prion therapies. Biochem Biophys Res Commun. 363: 95-100 ,
• Patton RL, Kalback WM, Esh CL, Kokjohn TA, Van Vickle GD, Luehrs DC, Kuo YM, Lopez J, Brune D, Ferrer I, Masliah B, Newel AJ, Beach TG, Castaño EM, Raw AE. (2006) Amyloid beta peptide remnants in AN-1792-immunized Alzheimer's disease patients: a biochemical analysis. At J Pathol. 169: 1048-63
• Phiel CJ, Wilson CA, Lee VM, Klein PS. (2003) GSK-3alpha regulates production of Alzheimer's disease amyloid-beta peptides. Nature. 423: 435-9 ,
• Porat Y, Abramowitz A, Gazit E. (2006) Inhibition of amyloid fibril formation by polyphenols: structural similarity and aromatic interactions as a common inhibition mechanism. Chem Biol Drug Des. 67: 27-37 ,
• Pfeifer, A., Eigenbrod, S., Al-Khadra, S., Hofmann, A., Mitteregger, G., Moser, M., Bertsch, U. Kretzschmar, H. (2006). Lentivector-mediated RNAi efficiently suppresses prion protein and prolongs survival of scrapie-infected mice. J Clin Invest. 116 :. 3204-10 ,
• Prior M, Lehmann S, Sy MS, Molloy B, McMahon HE (2007) Cyclodextrin inhibit replication of scrapie prion protein in cell culture. J Virol. 81 (20): 11195-207
• Priola SA, Raines A, Caughey WS. (2000) Porphyrin and phthalocyanine antiscrapie compounds. Science. 287: 1503-6 ,
• Ravikumar B, Vacher C, Berger Z, Davies JE, Luo S, Oroz LG, Scaravilli F, Easton DF, Duden R, O'Kane CJ, Rubinsztein DC. (2004) Inhibition of mTOR induces autophagy and reduced toxicity of polyglutamine expansions in fly and mouse models of Huntington's disease. Nat Genet. 36: 585-95 ,
• Riviere C, Richard T, Vitrac X, Mérillon JM, Valls J, Monti JP. (2007) New polyphenols active on beta-amyloid aggregation. Bioorg Med Chem Lett. In press
• Sarkar S, Davies JE, Huang Z, Tunnacliffe A, Rubin DC. (2007) trehalose, a novel mTOR-independent autophagic enhancer, accelerates the clearance of mutant huntingtin and alpha-synuclein. J Biol Chem. 282: 5641-52 ,
• Sciarretta KL, Gordon DJ, Meredith SC. (2006) Peptide-based inhibitors of amyloid assembly. Methods Enzymol. 413: 273-312 ,
• Sato T, Kienlen-Campard P, Ahmed M, Liu W, Li H, Elliott JI, Aimoto S, Constantinescu SN, Octave JN, Smith SO. (2006) Inhibitors of amyloid toxicity based on beta-sheet packing of Abeta40 and Abeta42. Biochemistry. 45: 5503-16 ,
• Serkedjieva J, Toshkova R, Antonova-Nikolova S, Stefanova T, Teodosieva A, Ivanova I. (2007) Effect of a plant polyphenol-rich extract on the lung protease activities of influenza virus-infected mice. Antivir Chem Chemother. 18: 75-82
• Shoval, H., Lichtenberg, D., Gazit, E. (2007). The molecular mechanism of anti-amyloid effects of phenols. Amyloid 14: 73-87
• Soto C, Kascsak RJ, Saborio GP, Aucouturier P, Wisniewski T, Prelli F, Kascsak R, Mendez E, Harris DA, Ironside J, Tagliavini F, Carp RI, Frangione B. (2000) Reversion of prion protein conformational changes by synthetic beta-sheet breaker peptides. 355: 192-7 ,
• Supattapone, S., Nguyen, H.B., Cohen, F.E., Prusiner, S.B., Scott, M.R. (1999). Elimination of prions by branched polyamines and implications for therapeutics. Proc Natl Acad Sci USA. 96: 14529-14534
• Tagliavini F, McArthur RA, Canciani B, Giaccone G, Porro M, Bugiani M, Lievens PM, Bugiani O, Peri E, Dall'Ara P, Rocchi M, Poli G, Forloni G, Bandiera T, Varasi M, Suarato A, Cassutti P, Cervini MA, Lansen J, Salmona M, Post C. (1997) Effectiveness of anthracycline against experimental prion disease in Syrian harnsters. Science. 276: 1119-22 ,
• Talhouk RS, Karam C, Fostok S, El-Jouni W, Barbour EK. (2007) Anti-inflammatory bioactivities in plant extracts. J Med Food. 10: 1-10 ,
• Wang J, Gines S, MacDonald ME, Gusella JF. (2005) Reversal of a full-length mutant huntingtin neuronal cell phenotype by chemical inhibitors of polyglutamine-mediated aggregation. BMC Neurosci 6: 1
• White AR, Enever P, Tayebi M, Mushen's R, Linehan J, Brandner S, Anstee D, Collinge J, Hawke S. (2003) Monoclonal antibodies inhibit prion replication and delay the development of prion disease. Nature. 422: 80-3
• Westerheide SD, Bosman JD, Mbadugha BN, Kawahara TL, Matsumoto G, Kim S, Gu W, Devlin JP, Silverman RB, Morimoto RI. (2004) Celastrols as inducers of the heat shock response and cytoprotection. J Biol Chem. 279: 56053-60
• White, AR, Enever, P., Tayebi, M., Mushens, R., Linehan, J., Brandner, S., Anstee, D., Collinge. J., Hawke, S. (2003). Monoclonal antibodies inhibit prion replication and delay the development of prion disease. Nature. 422: 80-3 ,
• Wu, JA, Attele, AS, Zhang, L., Yuan, CS (2001). Anti-HIV activity of medicinal herbs: usage and potential development. At J Chin Med. 29: 69-81 ,
• Yang F, Lim GP, Begum AN, Ubeda OJ, Simmons MR, Ambegaokar SS, Chef PP, Kayed R, Glabe CG, Frautschy SA, Cole GM. (2005) Curcumin inhibits formation of amyloid beta oligomers and fibrils, binds plaques, and reduces amyloid in vivo. J Biol Chem. 280: 5892-901 ,
• X, Smith DL, Meriin AB, Engemann S, Russell DE, Roark M, Washington SL, Maxwell MM, Marsh JL, Thompson LM, Wanker EE, Young AB, Housman DE, Bates GP, Sherman MY, Kazantsev AG. (2005) A potent small molecule inhibits polyglutamine aggregation in Huntington's disease neurons and suppresses neurodegeneration in vivo. Proc Natl Acad Sci USA 102: 892-7 ,
• Zhu M, Rajamani S, Kaylor J, Han S, Zhou F, Fink AL. (2004). The flavonoid baicalein inhibits fibrillation of alpha-synuclein and disaggregates existing fibrils. J Biol Chem. 279: 26846-57 ,

QUOTES INCLUDE IN THE DESCRIPTION

This list The documents listed by the applicant have been automated generated and is solely for better information recorded by the reader. The list is not part of the German Patent or utility model application. The DPMA takes over no liability for any errors or omissions.

Cited non-patent literature

• - Sciarretta et al 2006 [0009]
• Sato-T et al. 2006 [0009]
• Blanchard BJ et al. 2004 [0009]
• - Chirita et al. 2004 [0009]
• - Wang et al 2005 [0009]
• - Zhang, et al. 2005 [0009]
• - Desai et al. 2006 [0009]
• Bertsch et al., 2005 [0009]
• Johnson et al. 2005 [0010]
• - Beher et al. 2004 [0014]
• Phiel et al. 2003 [0014]
• - Netzer et al. 2003 [0014]
• - Best et al. 2007 [0014]
• Eriksen et al. 2003 [0014]
• - Cole al. 2006 [0014]
• - Hanessian et al. 2005 [0014]
• Bhat et al. 2003 [0015]
• Noble et al. 2005 [0015]
• - Yang et al. 2005 [0016]
• Matsubara et al. 2003 [0016]
• - Auluck et al. 2005 [0018]
• - Westerheide et al. 2004 [0018]
• Ravikumar et al. 2004 [0019]
• - Sarkar et al., 2007 [0019]
• - Gervasoni et al. 2004 [0021]
• Ouidja et al. 2007 [0021]
• McKenzie et al. 1994 [0021]
• Tagliavini et al. 1997 [0021]
• - Priola et al. 2000 [0021]
• - Soto et al. 2000 [0021]
• - Benito-Leon et al. 2004 [0021]
• - Korth et al. 2001 [0021]
• Supattapone et al. 2001 [0021]
• Gilch et al. 2001 [0021]
• - White et al. 2003 [0021]
• - Pfeifer et al. 2006 [0021]
• Caughey et al., 2003 [0021]
• - Prior et al. 2007 [0021]
• - Joshee et al. 2002 [0026]
• - Cushnie TP, & Lamb AJ. (2005) [0027]
• - Serkedjieva J et al. (2007) [0027]
• - Talhouk et al. (2007) [0027]
• - Li al (2007) [0027]
• - Porat et al. (2006), Rivière et al. (2006), Shoval et al. (2007) [0027]
• Kim et al. 2007 [0030]
• - Awadv et al. 2003 [0030]
• - Joshee et al., 2002 [0030]
• - Bomhoff et al. 2006 [0030]
• - Zhu et al. 2004 [0030]
• Kälvegren et al. 2007 [0030]
• - Ciesielska et al. 2002 [0031]
• - Chao et al. (2007) [0031]
• - Eiden et al. 2006 [0039]

Claims (15)

An agent for the prophylaxis and treatment of neurodegenerative protein deficiency and aggregation diseases, characterized in that they contain flavonoids as components active in vivo. Agent for the prophylaxis and treatment of neurodegenerative Protein deficiency and aggregation diseases, characterized that they are in identical or modified (eg, acetylation, Hydroxylation, halogenation, methylation, carbonylation, alkylation, Glycosylation, esterification, oxidation, hydrolysis, condensation, Polymerization) form synthetically produced flavonoids or their Become derivatives. Means according to claim 1, characterized in that the flavonoids out a) plants containing flavonoids or b) Sharing plants that contain flavonoids be won. Means according to claim 3, characterized in that the flavonoids from the herb or the root of plants, the flavonoids to be won. Agent according to claim 3 or 4, characterized that's it a) the plant Scutellaria (Skullcap) and / or b) around plants with structure-like flavonoids such as parsley, Paprika, chamomile, celery or chrysanthemums. Agent according to one of claims 1 to 5, characterized in that they a) the flavonoids Baicalein and / or Baicalin or b) structurally similar flavonoids from the group of flavones, such as acetaminophen, diurinary tangeretin, Luteolin, Apigenin or c) their derivatives or degradation products contain. Agent according to one of claims 1, 3, 4, 5 or 6, characterized in that they are individual substances, Mixtures, solutions or suspensions of flavonoids or Extracts of plants containing flavonoids. Means according to claim 7, characterized in that the extract by means of a) hot or cold water or b) alcohols or c) organic solvents, like eterners and / or d) maceration, dimacification, digestion, Re- / percolation, Soxhlet process, turbo (vortex), Ultra-Turrax, Ultrasonic and countercurrent extraction from plants that contain flavonoids contained, is won. Agent according to one of claims 1 to 8, characterized in that it is tea, available from Scutellaria. Agent according to one of claims 7 to 9, characterized in that the concentration of flavonoids in the Extract is in the range of 0.1 nM to 100 mM. Agent according to one of Claims 1 to 10, characterized in that the protein deficiency diseases are: a) Parkinson's disease, Lewy body dementia, multisystem atrophy b) Alzheimer's disease c) familial amyloidosis d) diabetes (type II) e) Huntington's disease f) spinocerebellar ataxia g) amyotrophic sclerosis h) tauopathies (Alzheimer's, frontotemporal dementia) i) senile amyloidosis j) Creutzfeldt-Jakob Disease, vCJD, GSS, FFI, Kuru Bovine Spongiform Encephalopathy, scrapie k) senile amyloidosis (ATTR amyloidosis). Process for the preparation of the agent according to of claims 2 to 9, characterized by the following steps: Pour 29 g of herb with 1 liter of boiling water, Let it draw for 10 minutes and strain. Use of the agents according to one of the claims 1 to 11 for the oral treatment of animals suffering from protein misfolding and single-agent, multiple or multiple diseases is carried out regularly. Use of the agents according to one of the claims 1 to 11 for the treatment of animals involved in protein misfolding and aggregation diseases, by intravenous, intramuscular, intraperitoneal, intraventricular and / or intracerebral administration, once, several times, or is carried out regularly. Use of the agents according to one of the claims 1 to 11 for the treatment of animals that respond to protein deficiency and aggregation diseases, via applicators, the one inhalable aerosol make the once, several times or is carried out regularly.