KR20070057779A - Compounds and their preparation for the treatment of alzheimer's disease by inhibiting beta-amyloid peptide production - Google Patents

Abstract

The present invention provides novel ginsenoside compounds, compositions (e.g. pharmaceutical compositions) comprising the ginsenoside compounds, and methods for the synthesis of these ginsenoside compounds. Additionally, the present invention provides methods for inhibiting beta-amyloid peptide production and methods for treating or preventing a pathological condition, particularly, neurodegeneration diseases (e.g. Alzheimer's disease), using these ginsenoside compounds.

Description

COMPOUNDS AND THEIR PREPARATION FOR THE TREATMENT OF ALZHEIMER'S DISEASE BY INHIBITING BETA-AMYLOID PEPTIDE PRODUCTION}

Cross Reference to Related Applications

This application was filed on October 7, 2004. U.S. Patent Application No. 10 / 961,346 and filed Jul. 16, 2004. We claim priority of provisional application 60 / 588,433

FIELD OF THE INVENTION

The present invention provides novel ginsenoside compounds, compositions (eg, pharmaceutical compositions) containing these ginsenoside compounds, and methods for synthesizing these ginsenoside compounds. Additionally, the present invention provides methods for inhibiting beta-amyloid peptide production using these ginsenoside compounds and methods for treating or preventing pathological abnormalities, in particular neurodegenerative diseases (eg, Alzheimer's disease).

Statement of Government Rights

In part, the present invention provides NIH Grant No. It was carried out with government support under ROIN543467. Accordingly, the US government reserves some rights in the present invention.

Alzheimer's disease (AD) is a neurodegenerative disorder characterized by a gradual loss of cognitive function that ultimately causes disability in normal social and / or occupational behavior (Francis, et al., Neuregulins and ErbB receptors in cultured neonatal astrocytes. J. Neurosci . Res . , 57: 487-94, 1999). Alzheimer's disease is the most common form of aging-related dementia and one of the most serious health problems in the United States. Alzheimer's disease, which affects approximately 4 million Americans, incurs at least $ 100 billion in treatment costs annually, making it the most expensive disease associated with aging. Alzheimer's disease occurs approximately twice as often in women as in men and accounts for more than 65% of elderly dementia. Alzheimer's disease is the fourth leading cause of death in the United States. To date, no cure for Alzheimer's disease is available and cognitive decline is inevitable. Although Alzheimer's disease may last as long as 20 years, AD patients live on average 8 to 10 years after being diagnosed with the disease.

The pathogenesis of Alzheimer's disease includes excessive amounts of neurofibrillary tangles (consisting of paired helical filaments and tau proteins) and elderly or senile spots (amyloid core) in the cerebral cortex. around the core is composed of neurites, astrocytes and glial cells. Geriatric spots and neurofibrillary tangles occur during normal aging, but are much more present in Alzheimer's patients. Specific protein malformations also appear in Alzheimer's disease. In particular, AD is characterized by deposition of amyloid β-peptide (Aβ) into amyloid plaque in the brain (Selkoe, et al. (2001) Alzheimer's disease: genes, proteins, and therapy.Physiol Rev. 81, 741-66; Hardy and Selkoe (2002). The amyloid hypothesis of Alzheimer's disease: progress and problems on the road to therapeutics. Science 297, 2209). Aβ is produced by sequential proteolytic cleavage of amyloid precursor protein (APP) by a group of membrane-bound proteases called β- and γ-secretases (Vassar and Citron (2000) Abeta-generating enzymes: recent advances in beta- and gamma-secretase research. Neuron 27, 419-422; John, et al. (2003) Human beta-secretase (BACE) and BACE inhibitors.J. Med Chem . 46, 4625-4630; Selkoe and Kopan (2003) Notch and Presenilin: regulated intramembrane proteolysis links development and degeneration.Annu . Rev Neurosci . 26, 565-597; Medina and Dotti (2003) ripped out by Presenilin-dependant gamma-secretase. Cell Signal 15, 829-841). Heterologous β-secretase cleavage at the C-terminus of Aβ yields two major isoforms of Aβ, Aβ40 and Aβ42. While Aβ40 is a predominant cleavage product, Aβ42, which is somewhat small but strongly induces amyloid production, is thought to be one of the key pathogenic factors in AD (Selkoe (2001) Alzheimer's disease: genes, proteins, and therapy.Physiol Rev. 18, 741-66), increased cerebral cortex Aβ42 is closely related to synaptic / nervous dysfunction associated with AD (Selkoe, Alzheimer's disease is a synaptic failure, Science 298, 789-791 (2002)).

Presenilin is required for intramembrane proteolysis of selected I-type membrane proteins, including amyloid-beta precursor protein (APP), to produce amyloid-beta proteins (De Strooper, . et al, Deficiency of presenilin- 1 inhibits the normal cleavage of amyloid precursor protein Nature 391:....... 387-90, 1998; Steiner and Haass, Intermembrane proteolysis by presenilins Nat Rev Mol Cell Biol 1: 217- 24, 2000; Ebinu and Yankner, A rip tide in neuronal signal transduction Neuron 34: 499-502, 2002; De Strooper and Annaert, Presenilins and the intramembrane proteolysis of proteins:... fact and fiction Nat Cell Biol . 3: E221-25, 2001; Sisodia and George-Hyslop, γ-Secretase, Notch, α-beta and Alzheimer's disease: where do the presenilins fit in? Nat . Rev. Neurosci . 3: 281-90, 2002). This proteolysis is mediated by the presenilin-dependent β-secretase mechanism, which is known to be highly conserved in all species, including nematodes, flies and mammals (L'Hernault and Arduengo, Mutation of a). .. sperm putative membrane protein in Caenorhabditis elegans sperm prevents differentiation but not its associated meiotic divisions J. Cell Biol 119:. 55-58, 1992; Levitan and Greenwald, Facilitation of lin-12-mediated signaling by sel-12, a Caenorhabditis . elegans S182 Alzheimer's disease gene Nature 377: 351-54, 1999; Li and Greenwald, HOP-1, a Caenorhabditis elegans presenilin, appears to be functionally redundant with SEL-12 presenilin and to facilitate LIN-12 and GLP-1 signaling. .... proc Natl Acad Sci USA 94:..... 12204-209, 1997; Steiner and Haass, intermembrane proteolysis by presenilin Nat Rev Mol Cell Biol 1:. 217-24, 2000; Sisodia and George-Hyslop, γ-Secretase, Notch, α-beta and Alzheimer's disease: wher ?.. e do the presenilins fit in Nat Rev Neurosci 3:. 281-90, 2000).

Γ-secretase, a high molecular weight multi-protein complex containing presenillin heterodimer and nicastrin, mediates the final stage of Aβ production in Alzheimer's disease (Li, et al., Presenilin ..... 1 is linked with β-secretase activity in the detergent solubilized state Proc Natl Acad Sci USA 97:. 6138-43, 2000; Esler, et al, Activity-dependent isolation of the presenilin-γ-secretase complex reveals .... nicastrin and a gamma substrate Proc Natl Acad Sci USA 99:. 2720-25, 2002). Presenilin heterodimers (converted from short-term pools to long-term pools) and other undefined core components appear to be critical for γ-secretase activity (Thinakaran, et al., Evidence that ... levels of presenilin (PS1 and PS2) are coordinately regulated by competition for limiting cellular factors J. Biol Chem 272:. 28415-422, 1997; Tomita, et al, the first proline of PALP motif at the C terminus of presenilins .. is obligatory for stabilization, complex formation, and gamma-secretase activities of presenilins J. Biol Chem 276:. 33273-281, 2001). γ-secretase activity shows very loose sequence specificity near the target transmembrane and cleavage site, and notch (Schroeter, EH, et al. (1998) Notch-1 signaling requires ligand-induced proteolytic release of intracellular domain ... Nature 393, 382-386; De Strooper, et al (1999) presenilin-1-dependent gamma-secretase-like protease mediates release of Notch intracellular domain Nature 398: 518-522), ErbB4 (Lee, et al. , (2002) Presenilin-dependent gamma -secretase-like intramembrane cleavage of ErbB4 J. Biol Chem 277, 6318-6323;.... Ni, et al (2001) Gamma-Secretase cleavage and nuclear localization of ErbB4 receptor tyrosine kinase. Science 294, 2179-2181), p75 neurotrophic factor receptor (p75NTR) (Jung, et al . (2003) Regulated intramembrane proteolysis of the p75 neurotrophin receptor modulates its association with the TrkA receptor. J, Biol. Chem. 278, Intramembrane cleavage of other non-APP I-type membrane substrates, including 42161-42169). It has been found that parameter. Global blockade of β-secretase activity not only abolishes Aβ development but also inhibits normal processing of these substrates, which is required for proper cellular function of other cellular β-secretase substrates. Thus, complete inhibition of γ-secretase activity can potentially cause serious side effects (Doerfler, et al., Links Free in PMC Presenilin-dependnet gamma-secretase activity modulates thymocyte development. (2001) Proc Natl . Acad . Sci USA 98, 9312-9317; Hadland, et al. Gamma-secretase inhibitors repress thymocyte development. Proc Natl. Acad . Sci USA 98, 7487-7491). A safer and more ideal approach is to use reactants that can selectively reduce A [beta] 42 production without affecting the intramembrane proteolysis of other γ-secretase substrates. For example, a subset of nonsteroidal anti-inflammatory drugs (NSAIDs) has little effect on the γ-secretase-mediated cleavage of ErbB4 (Weggon, et al. (2003) .Abeta42-lowering nonsteroidal anti-inflammatory drugs preserve intramembrane cleavage of the amyloid precursor protein (APP) and ErbB-4 receptor and signaling through the APP intracellular domain. J. Biol. Chem. 278, 30748-30754), was found to reduce Aβ42 generation (Weggon, et al. (2001). A subset of NSAIDs lower amyloidogenic Abeta42 independently of cyclooxygenase activity. Nature 414, 212-216). Thus, small molecules that can selectively inhibit A [beta] 42 production (without affecting the cleavage of other [gamma] -secretase substrates) are attractive and promising as therapeutic agents for treating AD.

Most cases of early-onset familial Alzheimer's disease (FAD) are caused by mutations in two related genes that encode presenilin proteins: PS1 and PS2 (Tanzi, et al., The .. gene defects responsible for familial Alzheimer 's disease Neurobiol Dis 3:... 159-68, 1996; Hardy, J. Amyloid, the presenilins and Alzheimer's disease Trends Neurosci 20: 154-59, 1997; Selkoe, DJ, Alzheimer's disease: . genes, proteins, and therapy Physiol Rev 81:.. 741-66, 2001). FAD-associated mutations in presenilins result in an amyloid-beta form (Aβ42) that is longer (42 amino acid residues) and more strongly induces amyloid production. The detoxification of pathobiology associated with presenilins offers a special opportunity to explain the molecular basis for Alzheimer's disease. Excessive beta-amyloid production is thought to cause neurodegeneration that is responsible for the dementia properties of AD.

Ginseng (ginseng) is the name for the dried roots of the Panax genus plant, which has been widely used in Asia for thousands of years as a medicine to treat health tonics and various diseases (Cho, et al. (1995). . (. eds) Korean ginseng In the Society for Korean ginseng: Understanding Korean ginseng, Seoul:. Hanlim Publishers, pp 35-54; Shibata S. (2001) Chemistry and cancer preventing activities of ginseng saponins and some related triterpenoid compounds J Korean Med Sci . 16 Suppl: S28-37; Attele, et al. (1999); Ginseng pharmacology: multiple constituents and multiple actions. Biochem Pharmacol . 58: 1685-1639; Coleman, et al. (2003). The effects of Panax ginseng on quality of life. J. Clin . Pharm. Ther . 28, 5-15; Coon and Ernst (2002). Panax ginseng: a systematic review of adverse effects and drug interactions. Drug Saf . 25: 323-44). The Panax genus includes approximately six species native to East Asia and two species native to Northeast America. Korean ginseng (Panax ginseng) and American ginseng (Panax quinquefolius L) are the two most commonly used species in health foods and pharmaceutical compositions. Their roots and extracts contain various substances, including saponins.

Ginseng is widely known to have specific pharmacological effects including anti-atherosclerosis, anti-thrombosis, anti-stress, anti-diabetic, anti-hypertensive, anti-tumor effects as well as improved liver function and immunity. Among the various types of compounds isolated from ginseng roots, ginseng saponins are known to be chemical components that contribute to their pharmacological effects. These compounds are triterpene glycosides named ginsenosides Rx (x being indices "a" to "K" depending on polarity). Polarity is determined by their mobility in thin-player chromatography plates and is a function of the total number of monosaccharide residues in the sugar chains of the molecule.

To date, at least 31 ginsenosides have been separated from white and red ginseng. All ginsenosides can be classified into three groups according to aglycon: protopanaxadiol-type ginsenosides (eg, Rb1, Rb2, Rc, Rd, (20R) Rg3, ( 20S) Rg3, Rh2), protopanaxatriol-type ginsenosides (eg Re, Rf, Rg1, Rg2, Rh1), oleanolic acid-type ginsenosides (eg Ro ). Protopanaxadiol- and protopanaxatriol-type ginsenosides have a triterpene skeletal structure known as dammarane (Attele, et al. (1999) Ginseng pharmacology :. multiple constituents and multiple actions Biochem Pharmacol 58:.. 1685-1693). Rk1, Rg5, (20R) Rg3, (20S) Rg3 are ginsenosides uniquely present in heat-treated ginseng but are not found as trace elements in untreated ginseng (Kwon, et al. (2001). .) Liquid chromatographic determination of less polar ginsenosides in processed ginseng J. Chromatogr A 921:....... 335-339; Park, et al (2002); Cytotoxic dammarane glycosides from processed ginseng Chem Pharm Bul 50, 538- . 540 Park, et al (2002 );.... Three new dammarane glycosides from heat-processed ginseng Arch Pharm Res 25, 428-432 Kim, et al (2000);. Steaming of ginseng at high temperature enhances biological activity. J. Nat . Prod . 63: 1702-1702). Carbohydrates, including glucopyranosyl, arabinopyranosyl, arabinofuranosyl, rhamnopyranosyl, can also be chemically associated with certain ginsenosides.

Treatment of ginseng with steam at high temperatures further enhances the content of these unique ginsenosides Rk1, Rg5, (20R) Rg3, (20S) Rg3, which appear to possess new pharmacological activity. At least some of the beneficial attributes of ginseng may be attributed to the content of triterpene saponins, which are mixtures of glucosides collectively referred to as ginsenosides.

U.S. Patent 5,776,460 describes processed ginseng products that exhibit enhanced pharmacological effects. These ginseng products, commercially known as “sun ginseng,” contain increased levels of active pharmacological components due to heat-treatment of ginseng for a certain time at high temperatures. As specifically described in US Pat. No. 5,776,460, the heat treatment of ginseng may be performed at 120 to 180 ° C. for 0.5 to 20 hours, preferably at 120 to 140 ° C. for 2 to 5 hours. The heating time depends on the heating temperature, where lower heating temperatures require longer heating times and higher heating temperatures require relatively short heating times. U. S. Patent No. 5,776, 460 also describes that processed ginseng products exhibit pharmacological properties, in particular anti-oxidant activity and vasodilation activity.

Recently, Kim Tae-Wan et al. Demonstrated that the unique components of the heat-treated ginseng products described in US Pat. No. 5,776,460 significantly lower Aβ42 production in cells (patent pending). Specifically, these inventors describe at least three ginsenosides Rk1, (20S) Rg3, Rg5 and (20S) Rg3, (20R) Rg3, which are unique components of heat-treated ginseng known as "sun ginseng". It was found that Rgk351, a mixture of Rg5, Rk1, lowers the production of Aβ 42 in mammalian cells. Rgk351 and Rkl are most effective at reducing Aβ42 levels. Furthermore, Rkl has been shown to inhibit Aβ42 production in a cell-free assay using partially purified γ-secretase complexes, indicating that Rkl is specific for the γ-secretase enzyme and / or Or, in addition, Kim Tae-Wan et al. Found that certain ginsenosides that do not exhibit Aβ42-reducing activity in vitro are effective in reducing Aβ42 in vivo. Some ginsenosides of the S) -protopanaxatriol (PPT) group, for example Rgl, can be converted to PPT after oral injection, thus Rg1 is in vitro amyloid-reducing activity Although not shown generally, it can be converted to the amyloid-reducing active compound PPT in vivo.

Summary of the invention

The present invention provides compositions and methods for preventing and treating neurodegenerative diseases such as Alzheimer's disease.

In one aspect, the present invention provides a compound having the formula:

R ₁ is selected from α-OH, β-OH, α-OX, β-OX, α-R ₆ COO-, β-R ₆ COO-, α-R ₆ PO _3- , β-R ₆ PO _3- Wherein X is a carbohydrate comprising one or more sugars or an acylated derivative thereof;

R ₆ is alkenyl, aryl, or alkyl I;

R ₂ is selected from H, OH, OAc, OX, wherein X is a carbohydrate comprising at least one sugar or an acylated derivative thereof;

R ₃ is selected from H, OH, OAc;

R ₄ is alkenyl, aryl, or alkyl II;

R ₅ is H or OH. The alkyl I group further carries oxygen, nitrogen, or phosphorus, and the alkyl II group further carries a functional group selected from hydroxyl, ether, ketone, oxime, hydrazone, imine, Schiff base. In one embodiment, the sugar group is selected from Glc, Ara (pyr), Ara (fur), Rha, Xyl. In other embodiments, R ₄ is selected from:

Any stereo-center arrangement is R or S;

X is OR or NR, wherein R is alkyl or aryl;

X 'is alkyl, OR, NR, wherein R is alkyl or aryl;

R 'is H, alkyl, or acyl. In another embodiment, the present invention provides a composition, in particular a pharmaceutical composition, containing a compound having the formula:

R ₁ is selected from α-OH, β-OH, α-OX, β-OX, α-R ₆ COO-, β-R ₆ COO-, α-R ₆ PO _3- , β-R ₆ PO _3- Wherein X is a carbohydrate comprising one or more sugars or an acylated derivative thereof;

R ₆ is alkenyl, aryl, or alkyl I;

R ₂ is selected from H, OH, OAc, OX, wherein X is a carbohydrate comprising one or more sugars or acylated derivatives thereof;

R ₃ is selected from H, OH, OAc;

R ₄ is alkenyl, aryl, or alkyl II;

R ₅ is H or OH.

The present invention also provides a method for synthesizing a compound having the formula wherein the method comprises the following steps:

(a)

Treating a compound having the formula with an oxidizing agent to form a compound having the formula

(b) treating the compound formed in step (a) with a reducing agent to form a compound having the formula

R ₁ is H or OH;

R ₂ is selected from H, OH, OAc, OX, wherein X is a carbohydrate comprising at least one sugar or an acylated derivative thereof;

R ₃ is selected from H, OH, OAc;

R ₄ is alkenyl, aryl, or alkyl. In one embodiment, the oxidant is chromic anhydride and the reducing agent is NaBH ₄ .

The present invention also provides a method for synthesizing a compound having the formula wherein the method comprises the following steps:

(a)

Treatment of a compound having the formula with an oxidant to form a compound having the formula

(b) treating the compound formed in step (a) with a reducing agent to form a compound having the formula

(c) optionally, treating the compound formed in step (b) with a protected R ₁ derivative to form a compound having the formula

(d) The compound formed in step (c) is treated with a deprotection agent to form a compound having the formula

R ₁ is selected from α-OH, β-OH, α-OX, β-OX, α-R ₆ COO-, β-R ₆ COO-, α-R ₆ PO _3- , β-R ₆ PO _3- Wherein X is a carbohydrate comprising one or more sugars or an acylated derivative thereof;

R ₆ is alkenyl, aryl, or alkyl I;

R ₂ is selected from H, OH, OAc, OX, wherein X is a carbohydrate comprising at least one sugar or an acylated derivative thereof;

R ₃ is selected from H, OH, OAc;

R ₄ is alkenyl, aryl, or alkyl II;

R ₅ is H or OH.

In addition, the present invention provides a method for synthesizing a compound having the formula wherein the method comprises the following steps:

(a)

Treatment of a compound having the formula with an oxidant to form a compound having the formula

(b) treating the compound formed in step (a) with a protecting agent to form a compound having the formula

(c) treating the compound formed in step (b) with a reducing agent to form a compound having the formula

(d) treating the compound formed in step (c) with Ac ₈ -Glc-Glc-Br to form a compound having the formula

(e) treating the compound formed in step (d) with a deprotecting agent to form a compound having the formula

(f) further modifying the compound formed in step (e) to form a compound having the formula:

In one embodiment, the starting material, betulafolienetriol, is obtained from a plant, for example a common birch.

In one aspect, the present invention provides a method for synthesizing a compound having the formula wherein the method comprises the following steps:

Compounds having the formula below are treated with a reducing agent such as NaBH ₄ :

In another aspect, the present invention provides a method for synthesizing a compound having the formula wherein the method comprises the following steps:

(a)

Treatment of a compound having the above formula with a reducing agent forms a compound having the following formula:

(b) treating the compound formed in step (a) with Ac ₈ -Glc-Glc-Br to form a compound having the formula

(c) treating the compound formed in step (b) with a deprotecting agent to form a compound having the formula

In addition, the present invention provides a method of treating or preventing pathological abnormalities in a subject, the method comprising administering to the subject a compound having the formula:

R ₁ is selected from α-OH, β-OH, α-OX, β-OX, α-R ₆ COO-, β-R ₆ COO-, α-R ₆ PO _3- , β-R ₆ PO _3- Wherein X is a carbohydrate comprising one or more sugars or an acylated derivative thereof;

R ₆ is alkenyl, aryl, or alkyl I;

R ₂ is selected from H, OH, OAc, OX, wherein X is a carbohydrate comprising at least one sugar or an acylated derivative thereof;

R ₃ is selected from H, OH, OAc;

R ₄ is alkenyl, aryl, or alkyl II;

R ₅ is H or OH. In one embodiment, the pathological abnormality is a neurodegenerative, preferably Alβheimer's disease, Aβ42-associated disease.

Furthermore, the present invention provides a method of inhibiting β-amyloid production in a subject, including inhibiting β-amyloid production in an in vitro background, the method comprising administering to the subject a compound having the formula Includes:

R ₁ is selected from α-OH, β-OH, α-OX, β-OX, α-R ₆ COO-, β-R ₆ COO-, α-R ₆ PO _3- , β-R ₆ PO _3- Wherein X is a carbohydrate comprising one or more sugars or an acylated derivative thereof;

R ₆ is alkenyl, aryl, or alkyl I;

R ₂ is selected from H, OH, OAc, OX, wherein X is a carbohydrate comprising at least one sugar or an acylated derivative thereof;

R ₃ is selected from H, OH, OAc;

R ₄ is alkenyl, aryl, or alkyl II;

R ₅ is H or OH.

Additional aspects of the invention will be apparent in light of the following detailed description.

1 shows sequential proteolytic processing of β-amyloid precursor protein (APP) mediated by β- and γ-secretase.

In Figure 2 (a) white ginseng; (b) red ginseng; (c) HPLC profile of ginseng (heat treated ginseng) is shown.

In FIG. 3, the chemical formulas of (a) Rg3, (b) Rk1 and (c) Rg5 are illustrated.

4 shows that Rgk351, (20S) Rg3, Rk1, Rg5 reduce the production of Aβ42 in CHO cells stably transfected with human APP695. CHO cells were treated with the indicated compounds (at 50 μg / ml) for 8 hours. Aβ42 levels in the medium were determined by ELISA and normalized to intracellular full-length APP.

5 shows that Rgk351, Rk1, Rg5 treatment reduces Aβ42 in a dose-dependent manner in the medium of CHO cells expressing human APP.

6 illustrates that Rgk351, Rk1, Rg5 treatment preferentially reduces Aβ42 (vs. Aβ40) in a dose-dependent manner in the medium of CHO cells expressing human APP. Relative levels of Aβ40 and Aβ42 were normalized to values obtained from untreated and vehicle-treated cells. Similar results were obtained using Neuro2a-sw (mouse Neuro2a cells expressing Swedish familial Alzheimer's disease variant forms of APP) and 293 cells expressing human APP.

Figure 7 shows the results of analysis of cell lysates and shows that Rgk351, Rk1, Rg5 induce increased accumulation of APP C-terminal fragments (γ-secretase substrate), while full-length holoAPP levels are not affected. .

In FIG. 8, Rgk351 and Rk1 treatment reduced Aβ42 levels in CHO cells co-expressing human APP with wild type presenilin 1 or familial Alzheimer's-associated variant forms of delta E9 ad L286V (delta E9 ad L286V). Illustrate that. The effect of Rg5 on Aβ42 production was much smaller than that of Rgk351 and Rk1.

9 shows the effect of Rk1 (R1) and Rg5 (R5) on Aβ42-specific γ-secretase activity. Naproxen (NP) and sulindac sulfide (SS) were simultaneously examined.

10 shows the effect of intrinsic ginsenosides on A [beta] 42 production. The structures of the seven standard ginsenosides (Rb1, Rb2, Rc, Rd, Re, Rg1, Rg2) investigated are shown in Table 1. CHO cells stably transfected with human APP695 with wild type of PSI (A, CHO-APP / PS1 cells) or ΔE9 FAD variant forms (B, CHO-APP / ΔE9PSI cells) were used. Cells were treated for 8 hours with the indicated compounds (at 50 μΜ). The levels of Aβ40 and Aβ42 secreted in the medium were determined by ELISA and normalized to intracellular full-length APP. In CHO-APP / PS1 cells, the average Aβ amount in the control sample was 320 pM for Aβ40 and 79 pM for Aβ42. Relative levels of Aβ40 and Aβ42 are normalized to values from untreated and vehicle-treated cells and expressed as% + s.d. for controls. One of three representative experiments is shown.

FIG. 11 shows the Aβ42-inhibitory activity of several ginsenosides derived from heat- or steam-treated ginseng. CHO-APP / PS1 (A) and CHO-APP / ΔE9PS1 (B) cells were treated with the indicated compounds (at 50 μM) for 8 hours, and the levels of secreted Aβ40 and Aβ42 were determined as described in FIG. 1. It was. The efficacy of the Aβ 42-reducing activity was in the order of Rk1> / = (20S) Rg3> Rg5> (20R) Rg3, and the effects of Rh1 and Rg6 were not significant. Rh2 also showed Aβ42-inhibitory effects, but cell viability was partially affected by 50 μM treatment (data not shown). PS1-ΔE9 FAD mutation reduced Aβ42 response to Rk1 treatment (B).

12 shows that Rgk351, Rk1, Rg5 treatment reduces Aβ42 in a dose-dependent manner in the medium of CHO-APP cells. (A) Dose-response of Aβ 42 -inhibiting activity of Rk1 and Rg5. IC50 of Rk1 was approximately 20 μΜ. (B) Rk1 preferentially inhibits Aβ42 (vs. Aβ40) in cultured CHO-APP cells, wherein the Aβ42-inhibition pattern of Rk1 is similar to the Aβ42-inhibition pattern of sulindac sulfide (SS). Relative levels of Aβ40 and Aβ42 were normalized to the values obtained in untreated and vehicle-treated cells. Similar results were obtained using Neuro2a-sw (mouse Neuro2a cells expressing Swedish familial Alzheimer's disease variant form of APP) and 293 cells expressing human APP (data not shown). The effect of Rg5 on Aβ42 production was much smaller than that of Rgk351 and Rk1.

13 shows the analysis result of APP processing after Rk1 processing. Steady-state levels of full-length APP and APP C-terminal fragments (APP-CTF) were examined by Western blot analysis using anti-R1 antibodies. Rgk351 (mixture of Rg3, Rg5, Rk1), Rk1, Rg5 treatments APP in mouse neuroblastoma neuro2a cells (KM670 / 671NL) stably expressing CHO-APP cells and Swedish FAD variant form (APPsw) of APP. Increased accumulation of C-terminal fragments (γ-secretase substrate) was induced. Correlated Aβ 42 levels for each sample are shown in the bottom panel.

In Figure 14, Aβ42-inhibited ginsenoside Rk1 is involved in the generation of intracellular domains (ICDs) from APP (A, AICD), Notch1 (B, NICD) or p75 neurotrophic factor receptors (p75NTR, p75-ICD). It shows no effect. Compound isolated and designated from 293 cells overexpressing APP (A), Notch-ΔE (B) or p75-ΔE (C): Compound E (CpdE, comprehensive γ-secretase inhibitor), Rgk351, Mkbrane fraction cultured in the presence of Rk1, sulfindac sulfide (SS). Only a small amount of AICD, NICD, p75-ICD was detected in control (-Incubate) or CpdE treated samples, while abundant production of AICD, NICD, p75-ICD was found in samples incubated with Rgk351, Rk1, and SS It became.

15 shows that Aβ42-inhibited ginsenosides Rk1 and (20S) Rg3 inhibit Aβ production in acellular γ-secretase assay. (A) CHAPSO-lysed membrane fractions were incubated with recombinant γ-secretase substrate and designated compounds (at 100 μM), and levels of Aβ42 and Aβ40 were determined by ELISA as described previously (27-29). . (B) Dose-response of Aβ40 and Aβ42-inhibitory activities of Rk1 and (20S) Rg3 on acellular γ-secretase assay. IC ₅₀ of Rk1 was 27 ± 3 μM for Aβ40 and 32 ± 5 μM for Aβ42. The IC ₅₀ of (20S) Rg3 was 27 ± 4 μM for Aβ40 and 26 ± 7 μM for Aβ42.

FIG. 16 shows the effect of two major metabolites of ginsenosides, including 20 (S) -protopanaxatriol (PPT) and 20 (S) -protopanaxadiol (PPD), on Aβ42 production. . 20 (S) -Panaxtriol (PT) and 20 (S) -Panaxadiol (PD) are artificial derivatives of PPT and PPD, respectively. PPT or PT treatment reduced Aβ 42 production without affecting the levels of Aβ 42 in Neuro2a cells (Neuro2a-SW, bottom panel) expressing human Swedish variant forms of APP and CHO cells expressing wild type human APP. (No data presented). PPD and PD did not give any inhibitory effect on Aβ40 or Aβ42 production.

Figure 17 shows the results of mass spectrometric analysis of Aβ species generated from CHO-APP cells treated with DMSO (carrier), Rk1 or (20S) Rg3. This treatment leads to a decrease in Aβ 42 species (1-42) and an elevation in Aβ 37 (1-37) and Aβ 38 (1-38). A:; (31894-31902 271 Wang R, Sweeny D, Gandy SE, Sisodia SS The profile of soluble amyloid β-protein in cultured cell media J. Bio Chem 1996....) Mass spectrometry analysis of Aβ species literature It was performed as described.

FIG. 18 shows the results of assayed Aβ levels secreted after treatment of CHO-APP cells with DMSO (Control 1), Naproxen (Control 2), Rk1 or (20S) Rg3. Αβ was immunoprecipitated with 4G8 antibody (Senetek), SDS-PAGE using tricine / urea gel (the protocol provided by Dr. Y. Ihara, University of Tokyo), 6E10 Analysis was performed by Western blot analysis using an antibody (Senetek). Synthetic Aβ40 and Aβ42 peptides were used to identify corresponding Aβ species.

19 shows the effect of ginsenosides Rk1 and (20S) Rg3 on Aβ40 and Aβ42 secretion in primary embryonic cortical neurons derived from Tg2576 transgenic mice. Rk1 and Rg3 treatment reduced the levels of secreted Aβ40 and Aβ42.

In this specification and the appended claims, the singular forms “a,” “an” and “the” include plural forms unless the context clearly dictates otherwise. Thus, by way of example, reference to a singular “drug” encompasses a plurality of drugs, and reference to a singular “ginsenoside” refers to one or more ginsenosides and their equivalents known to those skilled in the art. All publications, patent applications, in particular, and other references described herein are incorporated by reference in their entirety.

The present invention provides compounds and methods for treating Alzheimer's disease and neurodegeneration and for modulating the production of amyloid-beta protein (Aβ).

In one aspect, the present invention provides a compound having the formula:

R ₁ is selected from α-OH, β-OH, α-OX, β-OX, α-R ₆ COO-, β-R ₆ COO-, α-R ₆ PO _3- , β-R ₆ PO _3- Wherein X is a carbohydrate comprising one or more sugars or an acylated derivative thereof;

R ₆ is alkenyl, aryl, or alkyl I;

R ₂ is selected from H, OH, OAc, OX, wherein X is a carbohydrate comprising at least one sugar or an acylated derivative thereof;

R ₃ is selected from H, OH, OAc;

R ₄ is alkenyl, aryl, or alkyl II;

R ₅ is H or OH. The alkyl I group further carries oxygen, nitrogen, or phosphorus, and the alkyl II group further carries a functional group selected from hydroxyl, ether, ketone, oxime, hydrazone, imine, Schiff base. In one embodiment, the sugar is selected from Glc, Ara (pyr), Ara (fur), Rha, Xyl. In other embodiments, R ₄ is selected from:

Any stereo-center arrangement is R or S;

X is OR or NR, wherein R is alkyl or aryl;

X 'is alkyl, OR, NR, wherein R is alkyl or aryl;

R 'is H, alkyl, or acyl. In this specification, these compounds are dammar, in particular ginsenosides and analogs thereof. In this specification, these compounds are dammar, in particular ginsenosides and analogs thereof. As used herein, “ginsenosides” refers to specific compounds Ra1, Ra2, Ra3, Rb1, Rb2, Rb3, Rc, Rd, Re, Rf, Rg1, (20R) Rg2, (20S) Rg2, (20R) Rg3, (20S) Rg3, Rg5, Rg6, Rh1, (20R) Rh2, (20S) Rh2, Rh3, Rh4, (20R) Rg3, (20S) Rg3, Rk1, Rk2, Rk3, Rs1, Rs2, Rs3, Rs4, Rs5 , Rs6, Rs7, F4, Rgk351, Protoparanasadiol (PPD), Protopanaxatriol (PPT), DHPPD-I, DHPPD-II, DHPPT-I, DHPPT-II, Sun ginseng, White ginseng ( triterpene glycoside species, including butanol-soluble fractions of white ginseng or red ginseng, or analogs or homologs thereof. Ginsenosides of the present invention may be chemically linked to carbohydrates including glucopyranosyl, arabinopyranosyl, arabinofuranosyl, rhamnopyranosyl, and the like. The ginsenosides of the present invention may be isolated ginsenoside compounds or isolated and more synthesized ginsenosides. The isolated ginsenosides of the present invention can be further synthesized using processes including, but not limited to, thermal synthesis, photosynthesis, chemical synthesis, enzyme synthesis, or other synthetic procedures known to those skilled in the art.

Furthermore, the present invention provides a method for synthesizing a compound having the formula wherein the method comprises the following steps:

(a)

Treating a compound having the formula with an oxidizing agent to form a compound having the formula

(b) treating the compound formed in step (a) with a reducing agent to form a compound having the formula

R ₁ is H or OH;

R ₂ is selected from H, OH, OAc, OX, wherein X is a carbohydrate comprising at least one sugar or an acylated derivative thereof;

R ₃ is selected from H, OH, OAc;

R ₄ is alkenyl, aryl, or alkyl. In one embodiment, the oxidant is chromic anhydride and the reducing agent is NaBH ₄ .

Starting materials, ie compounds having the formula: in particular, betulafolienetriol, are obtained from plants, including conventional birch trees:

Extracts of these plants are a preferred starting material for preparing ginsenosides because they are rich in betulapolyentriol and are much cheaper than ginseng.

The present invention also provides a method for synthesizing a compound having the formula wherein the method comprises the following steps:

(a)

Treatment of a compound having the formula with an oxidant to form a compound having the formula

(b) treating the compound formed in step (a) with a reducing agent to form a compound having the formula

(c) optionally, treating the compound formed in step (b) with a protected R ₁ derivative to form a compound having the formula

(d) The compound formed in step (c) is treated with a deprotection agent to form a compound having the formula

R ₁ is selected from α-OH, β-OH, α-OX, β-OX, α-R ₆ COO-, β-R ₆ COO-, α-R ₆ PO _3- , β-R ₆ PO _3- Wherein X is a carbohydrate comprising one or more sugars or an acylated derivative thereof;

R ₆ is alkenyl, aryl, or alkyl I;

R ₂ is selected from H, OH, OAc, OX, wherein X is a carbohydrate comprising at least one sugar or an acylated derivative thereof;

R ₃ is selected from H, OH, OAc;

R ₄ is alkenyl, aryl, or alkyl II;

R ₅ is H or OH. Alkyl I groups further carry oxygen, nitrogen, or phosphorus; Alkyl II groups further carry a functional group selected from hydroxyl, ether, ketone, oxime, hydrazone, imine, Schiff base. In one embodiment, the oxidant is chromic anhydride and the reducing agent is NaBH ₄ . In other embodiments, the protected R ₁ derivative is a protected R ₁ halogen derivative. For example, protected R ₁ derivatives are protected with Ac ₈ -groups. Protected R ₁ groups are deprotected using drugs such as NaOMe.

In addition, the present invention provides a method for synthesizing a compound having the formula wherein the method comprises the following steps:

(a)

Treatment of a compound having the formula with an oxidant to form a compound having the formula

(b) treating the compound formed in step (a) with a protecting agent to form a compound having the formula

(c) treating the compound formed in step (b) with a reducing agent to form a compound having the formula

(d) treating the compound formed in step (c) with Ac ₈ -Glc-Glc-Br to form a compound having the formula

(e) treating the compound formed in step (d) with a deprotecting agent to form a compound having the formula

(f) further modifying the compound formed in step (e) to form a compound having the formula:

In one embodiment, the oxidant is chromic anhydride, the reducing agent is NaBH ₄ , and the compound is deprotected with NaOMe.

The present invention also provides a method for synthesizing a compound having the formula wherein the method comprises the following steps:

Compounds having the formula below are treated with a reducing agent such as NaBH ₄ :

The present invention also provides a method for synthesizing a compound having the formula wherein the method comprises the following steps:

(a)

Treatment of a compound having the above formula with a reducing agent forms a compound having the following formula:

(b) treating the compound formed in step (a) with Ac ₈ -Glc-Glc-Br to form a compound having the formula

(c) treating the compound formed in step (b) with a deprotecting agent to form a compound having the formula

In one embodiment, the reducing agent is NaBH ₄ and the compound is deprotected with NaOMe.

In addition, the present invention is directed to the use of isolated or isolated and more synthesized ginsenosides to modulate amyloid-beta production in individuals, to treat or prevent Alzheimer's disease, to treat or prevent neurodegeneration. A ginsenoside composition containing a mixture is provided wherein at least one ginsenoside is selected from Ra1, Ra2, Ra3, Rb1, Rb2, Rb3, Rc, Rd, Re, Rf, Rg1, (20R) Rg2, (20S) Rg2. , (20R) Rg3, (20S) Rg3, Rg5, Rg6, Rh1, (20R) Rh2, (20S) Rh2, Rh3, Rh4, (20R) Rg3, (20S) Rg3, Rk1, Rk2, Rk3, Rs1, Rs2 , Rs3, Rs4, Rs5, Rs6, Rs7, F4, Rgk351, protopanaxadiol, protopanaxatriol, DHPPD-I, DHPPD-II, DHPPT-I, DHPPT-II, sun ginseng ( sun ginseng, butanol-soluble fractions of white ginseng or red ginseng, or analogs or analogs thereof. In one embodiment of the invention, the ginsenoside composition is Rgk351.

The present invention provides methods and pharmaceutical compositions used to reduce amyloid-beta production, which methods include the use of pharmaceutically acceptable carriers and ginsenoside compounds. Examples of pharmaceutically acceptable carriers, formulations of pharmaceutical compositions, and methods of preparing these formulations are described herein. These pharmaceutical compositions are useful for administering the dhamranane and ginsenoside compounds of the invention to a subject for the treatment of various diseases, including neurodegeneration and / or symptoms associated therewith, as described herein. Ginsenoside compounds are provided in an amount effective to treat a disease (eg, neurodegeneration) in an individual to which such pharmaceutical composition is administered. As noted above, one skilled in the art can conveniently determine an appropriate amount. In one embodiment, the present invention provides a method of inhibiting β-amyloid production in a subject, the method comprising administering to the subject a compound having the formula:

R ₁ is selected from α-OH, β-OH, α-OX, β-OX, α-R ₆ COO-, β-R ₆ COO-, α-R ₆ PO _3- , β-R ₆ PO _3- Wherein X is a carbohydrate comprising one or more sugars or an acylated derivative thereof;

R ₆ is alkenyl, aryl, or alkyl I;

R ₂ is selected from H, OH, OAc, OX, wherein X is a carbohydrate comprising at least one sugar or an acylated derivative thereof;

R ₃ is selected from H, OH, OAc;

R ₄ is alkenyl, aryl, or alkyl II;

R ₅ is H or OH. As used herein, “individual” encompasses, for example, animals, in particular humans, mice, mice, rabbits, dogs, sheep, cattle, and in vitro systems, in particular cultured cells, tissues, organs.

The present invention also provides a method of treating neurodegeneration in a subject, which method reduces amyloid-beta production in the cell, and thus an effective amount of a ginsenoside compound or composition for treating neurodegeneration and the cells of the subject ( Preferably, CNS cells). Examples of neurodegeneration that can be treated with the methods of the invention include, but are not limited to, Alzheimer's disease, Amyotrophic axon sclerosis (Lou Gehrig's disease), Binswanger's disease, corticobasal degeneration (CBD), and obvious histopathology. Dementia lacking distinctive histopathology (DLDH), frontotemporal dementia (FTD), Huntington's chorea, multiple sclerosis, myasthenia gravis, Parkinson's disease (DLDH) Parkinson's disease, Pick's disease, progressive supranuclear palsy (PSP), and the like. In a preferred embodiment of the invention, the neurodegeneration is Alzheimer's disease (AD) or sporadic Alzheimer's disease (SAD). In another preferred embodiment of the invention, Alzheimer's disease is early-onset familial Alzheimer's disease (FAD). One skilled in the art can conveniently determine when clinical symptoms of neurodegeneration are alleviated or reduced.

The present invention also provides a method of treating or preventing pathological abnormalities such as neurodegeneration and Aβ42-related diseases in a subject, wherein the method comprises administering at least one ginsenoside compound to the subject in an amount effective to treat neurodegeneration. Administering. Aβ42-related diseases are any diseases caused by Aβ42 or exhibit symptoms of abnormal Aβ42 accumulation. As used herein, “effective for treating neurodegeneration” means effective for alleviating or minimizing clinical damage or symptoms of neurodegeneration. For example, if neurodegeneration is Alzheimer's disease, clinical damage or symptoms of neurodegeneration may result in progressive production of cognitive function by reducing amyloid-beta production and the development of senile plaques and neurofibrillary tangles. It can be mitigated or minimized by minimizing or weakening loss. The amount of inhibitor that is effective for treating neurodegeneration in an individual depends on the specific factors of each case, including the type of neurodegeneration, the stage of neurodegeneration, the weight of the individual, the severity of the individual's condition, and the method of administration. Effective amounts can be conveniently determined by one skilled in the art. In one embodiment, the present invention provides a method of treating or preventing neurodegeneration in a subject, the method comprising administering to the subject a compound having the formula:

R ₁ is selected from α-OH, β-OH, α-OX, β-OX, α-R ₆ COO-, β-R ₆ COO-, α-R ₆ PO _3- , β-R ₆ PO _3- Wherein X is a carbohydrate comprising one or more sugars or an acylated derivative thereof;

R ₆ is alkenyl, aryl, or alkyl I;

R ₂ is selected from H, OH, OAc, OX, wherein X is a carbohydrate comprising at least one sugar or an acylated derivative thereof;

R ₃ is selected from H, OH, OAc;

R ₄ is alkenyl, aryl, or alkyl II;

R ₅ is H or OH.

In one embodiment of the invention, Alzheimer's disease is treated in a subject by administering to the subject a therapeutically effective amount of a ginsenoside composition, ginsenoside, or analog or homologue thereof effective to treat Alzheimer's disease. Suitably the subject is a mammal (eg, human, pet and commercial animals, including cattle, dogs, monkeys, mice, pigs, mice), preferably humans. As used herein, an analog refers to chemical compounds that are structurally similar to other compounds and that can be derived from them but differ slightly in composition. For example, analogs of ginsenoside (20S) Rg3 are slightly different from (20S) Rg3 (eg, substitution of one atom by atoms of different elements or the presence of certain functional groups) but can be derived from (20S) Rg3 to be. As used herein, a homologue means a member of a series of compounds in which each member is distinguished from another member by one constant chemical unit. Synthesis herein means the formation of certain chemical compounds from constituent parts, using synthetic procedures known in the art. This synthesis process involves, for example, the use of thermal means, light means, chemical means, enzymatic means or other means of forming a particular chemical composition.

As used herein, a “therapeutically effective amount” or “effective amount” is a single dose or multiple doses, which is necessary to prevent, treat, alleviate or at least minimize clinical damage, symptoms or complications associated with Alzheimer's disease. Means the amount of the composition according to the invention. The amount of ginsenosides effective for treating Alzheimer's disease depends on the specific factors of each case, including the stage or severity of Alzheimer's disease, the weight of the subject, the condition of the subject, and the method of administration. One skilled in the art can conveniently determine these amounts. For example, clinical damage or symptoms of Alzheimer's disease may reduce dementia or other discomfort experienced by the individual; Prolong the survival of the individual beyond the life expectancy in the absence of such treatment; Or is alleviated or minimized by inhibiting or preventing the progression of Alzheimer's disease.

Treatment of Alzheimer's disease herein means, without limitation, treating one or more abnormalities causing Alzheimer's disease, including neurodegeneration, senile plaques, neurofibrillary concentrates, neurotransmitter deficiencies, dementia, and old age. In the present specification, the prevention of Alzheimer's disease prevents the onset of Alzheimer's disease, delays the onset of Alzheimer's disease, prevents the progress or progress of Alzheimer's disease, slows the progress or progress of Alzheimer's disease, and the progress or progress of Alzheimer's disease. Means to delay.

Prior to the present invention, the effects of dammaran and ginsenosides on the production of beta amyloid proteins are unknown. The present invention establishes that ginsenosides such as (20S) Rg3, Rk1, Rg5, or analogs or analogs thereof may also be used to prevent and treat Alzheimer's disease patients. These new therapies offer unique strategies to treat and prevent neurodegeneration and dementia associated with Alzheimer's disease by regulating the production of Aβ42. Furthermore, neurodegeneration and dementia not associated with Alzheimer's disease can also be treated or prevented by the ginsenosides of the invention that regulate the production of Aβ 42.

Ginsenosides of the present invention include natural or synthetic functional variants possessing ginsenoside biological activity and fragments of ginsenosides having ginsenoside biological activity. As used herein, “ginsenoside biological activity” refers to an activity that modulates the production of Aβ42, a 42 amino acid equivalent of amyloid β-peptide, which strongly shows amyloid production. In one embodiment of the invention, ginsenosides reduce the production of Aβ 42 in the cells of the individual. The well known ginsenosides and ginsenoside compositions are Ra1, Ra2, Ra3, Rb1, Rb2, Rb3, Rc, Rd, Re, Rf, Rg1, (20R) Rg2, (20S) Rg2, (20R) Rg3, (20S Rg3, Rg5, Rg6, Rh1, (20R) Rh2, (20S) Rh2, Rh3, Rh4, (20R) Rg3, (20S) Rg3, Rk1, Rk2, Rk3, Rs1, Rs2, Rs3, Rs4, Rs5, Rs6 , Rs7, F4, Rgk351, Protoparnacindiol (PPD), Protoparnacintriol (PPT), DHPPD-I, DHPPD-II, DHPPT-I, DHPPT-II, sun ginseng, white ginseng Or ginsenosides, including but not limited to butanol-soluble fractions of red ginseng, or analogs or homologs thereof. In one embodiment of the invention, the ginsenoside is Rk1. In another embodiment of the invention, the ginsenoside is (20S) Rg 3. In another embodiment, the ginsenoside is Rg 5. In another embodiment, the ginsenoside composition is Rgk351, which is a mixture of (20S) Rg3, Rg5, Rk1.

Ginsenosides such as Rk1, (20S) Rg3, Rg5 and methods for preparing analogs and homologues thereof are well known in the art. For example, US Pat. No. 5,777,460 describes a process for preparing processed ginseng products in which the ratio of ginsenosides (Rg3 + Rg5) to (Rc + Rd + Rb1 + Rb2) is greater than 1.0. The processing product described in US Pat. No. 5,776,460 is prepared by heat-treating ginseng at a high temperature of 120-180 ° C. for 0.5-20 hours. Ginsenosides of the present invention are isolated ginsenoside compounds or isolated and more synthesized ginsenoside compounds. The isolated ginsenosides of the present invention can be further synthesized using processes including, but not limited to, thermal synthesis, photosynthesis, chemical synthesis, enzyme synthesis, or other synthetic procedures known to those skilled in the art.

In the methods of the invention, the ginsenoside compound is administered to the subject in combination with one or more different ginsenoside compounds. Administration of the ginsenoside compound “cooperatively” with one or more different ginsenoside compounds means co-administration of these therapeutic agents. Co-administration can proceed simultaneously, sequentially, or alternately. Simultaneous co-administration means administration of different ginsenoside compounds at essentially the same time point. For simultaneous co-administration, the treatment may be carried out simultaneously with two or more different ginsenosides. For example, a single integrated formulation containing both an amount of a particular ginsenoside compound and an amount of a second, different ginsenoside compound in physical contact with each other can be administered to the subject. A single integrated formulation may consist of an oral formulation containing an amount of both ginsenoside compounds that can be administered orally to an individual, or a liquid mixture containing an amount of both ginsenoside compounds that can be injected into an individual.

In addition, in the present invention, an amount of a particular ginsenoside compound and an amount of one or more different ginsenoside compounds may be simultaneously administered to the individual in separate and independent formulations. Thus, the method of the present invention is not limited to simultaneous co-administration of different ginsenoside compounds in physical contact with each other.

In the methods of the present invention, the ginsenoside compound may be co-administered to the subject in separate independent agents spaced at regular time intervals to achieve the maximum efficacy of this combination. Administration of each therapeutic agent can be varied for a duration of time from short and rapid to continuous perfusion. In the case of spaced apart at regular time intervals, co-administration of the ginsenoside compound may be performed sequentially or alternately. For sequential co-administration, one of the therapeutic agents is administered separately following the other therapeutic agent. For example, the entire course of Rg5 derivative treatment is completed, followed by the entire course of Rk1 derivative treatment. Alternatively, for sequential co-administration, the entire course of Rk1 derivative treatment is completed, followed by the entire course of Rg5 derivative treatment. For alternative co-administration, a partial course of Rk1 derivative treatment is performed alternately with a partial course of Rg5 derivative treatment until the entire treatment of each therapeutic agent is completed.

The therapeutic agents of the present invention (ie, ginsenosides and analogs thereof) may be administered by known procedures including oral administration, parenteral administration (eg, intramuscular, intraperitoneal, endovascular, intravenous or subcutaneous administration), transdermal administration. Or to an animal subject. Suitably, the therapeutic agents of the invention are administered orally or intravenously.

For oral administration, the ginsenoside formulation may be provided in capsules, tablets, powders, granules, or suspensions. These formulations may contain conventional additives such as lactose, mannitol, corn starch or potato starch. These formulations may also be provided with binders such as crystalline cellulose, cellulose analogs, acacia, corn starch or gelatin. In addition, these agents may be provided with disintegrators such as corn starch, potato starch, or sodium carboxymethyl cellulose. These formulations may also be provided with dibasic calcium phosphate or anhydrous sodium starch glycolate. Finally, these formulations may be provided with lubricants, such as talc or magnesium stearate.

For parenteral administration, the ginsenoside preparation may be mixed with a sterile aqueous solution, preferably with a sterile aqueous solution that is isotonic with the blood of the individual. These formulations produce aqueous solutions by dissolving the solid active ingredient in water containing a physiologically-compatible substance such as sodium chloride, glycine and the like and having a buffered pH suitable for physiological conditions, and the solution It can be prepared by making this sterile. These formulations may be provided in unit or multi-dose containers, such as sealed ampoules or vials. In addition, these agents may be used without limitation, in episcial, intracapsular, intracutaneous, intramuscular, intraorbital, intraperitoneal (especially for topical site treatment). E), intraspinal, intrasternal, intravascular, intravenous, parenchymatous or subcutaneous.

For transdermal administration, ginsenoside preparations may be mixed with skin penetration enhancers such as propylene glycol, polyethylene glycol, isopropanol, ethanol, oleic acid, N-methylpyrrolidone, etc. increase permeability and allow the treatment to penetrate the bloodstream through the skin. The therapeutic / enhancer composition may be further mixed with a polymerizable material, such as ethylcellulose, hydroxypropyl cellulose, ethylene / vinylacetate, polyvinyl pyrrolidone, and the like to provide a gel form composition, wherein the composition is methylene chloride It may be dissolved in a solvent such as, evaporated to the desired viscosity, and then applied to a backing material to provide a patch.

Ginsenosides of the present invention may be released or delivered from an osmotic mini-pump. The release rate from the basal osmotic mini-pump can be controlled with a microporous fast-response gel of microporous structure disposed at the release orifice. Osmotic mini-pumps are useful for controlling the release of a therapeutic or for targeting delivery.

Ginsenoside formulations are further combined with pharmaceutically acceptable carriers, and therefore the present invention includes pharmaceutical compositions. Pharmaceutically acceptable carriers should be “acceptable” in that they are compatible with the other ingredients of the composition and should not be harmful to their recipients. Examples of acceptable pharmaceutical carriers are, in particular, carboxymethyl cellulose, crystalline cellulose, glycerin, gum arabic, lactose, magnesium stearate, methyl cellulose, powders, saline, sodium alginate, sucrose, starch, talc, water and the like. Formulations of pharmaceutical compositions may conveniently be presented in unit dosage form.

The formulations of the present invention can be prepared by methods well known in the pharmaceutical art. For example, the active compound may be combined with a carrier or diluent as a suspension or solution. Optionally, one or more accessory ingredients (eg, buffers, flavors, surface active agents, etc.) may also be added. The choice of carrier depends on the route of administration. Such pharmaceutical compositions are useful for administering to a subject a therapeutic agent of the invention (ie, ginsenosides and analogs thereof in a separate independent formulation or in a single integrated formulation) to treat Alzheimer's disease. The therapeutic agent is provided in an amount effective to treat or prevent Alzheimer's disease in a subject. These amounts can be conveniently determined by one skilled in the art.

The therapeutically effective amount of ginsenoside depends on the specific factors of each case, including the stage of Alzheimer's disease, the weight of the subject, the severity of the subject's condition, and the method of administration. For example, (20S) Rg 3 may be administered at a dose of approximately 5 μg / day to 1500 mg / day. Suitably, (20S) Rg3 is administered at a dose of approximately 1 mg / day to 1000 mg / day. Rg5 may be administered at a dose of approximately 5 μg / day to 1500 mg / day, preferably at a dose of approximately 1 mg / day to 1000 mg / day. Rk1 may be administered at a dose of approximately 5 μg / day to 1500 mg / day, preferably at a dose of approximately 1 mg / day to 1000 mg / day. Furthermore, the ginsenoside composition Rgk351 may be administered at a dose of about 5 μg / day to 1500 mg / day, preferably about 1 mg / day to 1000 mg / day. Appropriate therapeutically effective amounts of any particular ginsenoside compound within the stated ranges may be conveniently determined by one skilled in the art depending on the specific factors in each case.

Additionally, the present invention includes a method for preventing Alzheimer's disease in a subject with pre-Alzheimer's disease, the method comprising administering to the subject a therapeutically effective amount of a ginsenoside compound. do. As used herein, the term "pre-Alzheimer's disease" means a condition prior to the onset of Alzheimer's disease. Individuals with pre-Alzheimer's disease have not been diagnosed with Alzheimer's disease but have some of the typical symptoms of Alzheimer's disease and / or have a medical history that increases the risk of developing Alzheimer's disease. .

The present invention also provides a method of treating or preventing Alzheimer's disease in a subject, the method comprising administering to the subject a therapeutically effective amount of a ginsenoside compound.

The following examples illustrate the invention and are described to aid the understanding of the invention, but in no way limit the scope of the invention as defined by the following claims.

We believe that at least three ginsenoside compounds, Rk1, (20S) Rg3, Rg5 and mixture Rgk351, inhibit the production of Aβ42 in cells to treat non-AD-associated neuropathogenesis and / or AD and It was unexpectedly found to prevent the progression of non-AD related neuropathies. Rgk351 and Rk1 were most effective at reducing Aβ42 levels. Furthermore, Rk1 has been shown to inhibit Aβ42 production in a cell-free assay using partially purified γ-secretase complexes, indicating that Rk1 inhibits the specificity and / or specificity of the γ-secretase enzyme. Imply that it regulates activity.

Example  One

The potential effects of ginsenosides and their analogues on the treatment of AD were investigated. First, a number of ginsenosides were screened based on the effect on Aβ production. The effects of various ginsenosides on the production of Aβ (eg, Aβ40 and Aβ42) are, firstly, Chinese hamsters expressing human APP, with each ginsenoside purified from raw ginseng (“white ginseng”). Access was made by culturing ovary (CHO) cells (CHO-APP cells). These representative ginsenosides are Rb1, Rb2, Rc, Rd, Re, Rg1, Rg2, which are distinguished in the side chain and sugar moiety.

Table 1-3 shows the structure of ginsenosides used in this study and their effects on A [beta] 42 production. They are distinguished from two or three side chains attached to a common triterpene skeleton known as dammarane. Common structural skeletons for each group of ginsenosides are shown in the top panel. Ginsenosides with Aβ 42 -inhibiting activity are indicated in the rightmost column of the table: Aβ 42-inhibiting activity (“Yes”), no effect (“No”), not measured (“ND”). Ginsenosides that affect cell viability are labeled "Cytotoxic". Abbreviations for carbohydrates are as follows: Glc, D-glucopyranosyl; Ara (pyr), L-arabinopyranosyl; Ara (fur), L-arabinofuranosyl; Rha, L-Rhamnopyranosyl.

After 8 hours of incubation, the medium was collected and the levels of secreted Aβ40 and Aβ42 were determined by ELISA. Ginsenosides from Rb1, Rb2, Rc, Rd, Re, Rg1, Rg2 showed no inhibitory effect on Aβ40 and Aβ42 production (FIG. 10).

Steaming ginseng at high temperatures produced additional ginsenosides with enhanced pharmacological activity, including (20S) Rg3, Rk1, Rg5 (22-25). The effects of these heat-treated derived ginsenosides (eg, (20S) Rg3, Rh1, Rh2, Rk1, Rg6, Rg5) on Aβ40 and Aβ42 production were investigated. In the initial screening, three structurally related ginsenosides, Rk1, (20S) Rg3, Rg5, which selectively inhibit the secretion of Aβ 42, were identified (FIG. 11). In contrast, Aβ42 levels were not affected by (20S) Rg3, Rh1, Rg6. Aβ40 levels were not changed by treatment with any ginsenosides tested. The efficacy of Aβ42-inhibiting activity was highest in Rk1 and (20S) Rg3. Rg5 was less potent as Aβ42-inhibiting reactants compared to Rk1 or (20S) Rg3 (FIG. 2). Secretion of Aβ40 was affected by Rk1 treatment only at very high concentrations (~ 100 μM), and cell viability was not affected by Rk1 treatment under these conditions (up to 100 μM, 8 h treatment) (data not shown). . Interestingly, the PS1 ΔE9 FAD mutation (FIG. 11B) reduced Aβ42-inhibiting response to (20S) Rg3, Rk1, Rg5 treatment compared to PS1 wild type expressing cells (FIG. 11A). In further analysis, Rk1 and Rg5 were found to inhibit Aβ42 in a dose-dependent manner (FIG. 12A). Overnight treatment with Rgk351, Rk1, Rg5 also reduced Aβ42 production in CHO-APP cells (FIG. 12B). The Aβ 42 -inhibitory activity of Rk1 was similar to the Aβ 42 -inhibitory activity of sulindac sulfide, one of the known Aβ 42 -inhibited NSAIDs. During overnight treatment, A [beta] 40 production was also slightly affected by Rk1 or sulfindac sulfide treatment (FIG. 12B). These studies support the structure-activity correlation between ginsenoside chemical structure and Aβ42-inhibiting activity, and also provide the basis for designing additional Aβ42-inhibiting analogs and defining the class of compounds with Aβ42-inhibiting activity. do.

Rk1 did not affect the steady-state levels of full-length APP in both CHO-APP and Neuro2a-APPsw cells (FIG. 13), indicating that a decrease in Aβ42 resulted in modified post-translation of APP. implied likely due to processing). In contrast to full length morphology, the steady-state level of the C-terminal APP fragment was up-regulated by Rk1 treatment (FIG. 13). These data indicate that Rk1 affects the γ-secretase cleavage step (eg, Aβ42 cleavage), leading to the accumulation of APP C-terminal fragments, as observed in the comprehensive γ-secretase inhibitor Compound E. Hints. Aβ42 levels in the media of each corresponding sample are shown in the bottom panel.

Because the effects of Rk1 are highly selective for Aβ42 (not selective for Aβ40) in cell-based assays, the development of AICD is a transmembrane cleavage of APP distal from the Aβ40 or Aβ42 sites and in Notch1 or p75NTR cells. Rk1 differs from other γ-secras, including whether it is due to γ-secretase-mediated intramembrane cleavage of Notch1 or p75 neurotrophic factor receptor (p75NTR) yielding domains (NICD or p75-ICD, respectively). It was investigated whether it affected the lipase-mediated cleavage phenomenon. Cell-free generation of AICD, NICD, p75-ICD was not affected by incubation with Rgk351 or Rk1 (FIG. 5). Under these conditions, Compound E effectively inhibited the acellular development of ICD, and sulfindax sulfide did not affect ICD development from APP, Notch1 or p75NTR. These data suggest that Rk1 is not a comprehensive inhibitor of γ-secretase cleavage and does not affect intramembrane cleavage of other γ-secretase substrates such as Notch1 or p75NTR.

The inhibitory effect of Rk1 and (20S) Rg3 on A [beta] production was then examined in an in vitro [gamma] -secretase test. Both Rk1 and sulindac sulfide potentially inhibited Aβ42 production in vitro (FIG. 15). In contrast, naproxen, an NSAID without Aβ42-inhibiting activity, did not have any effect on Aβ42 production (FIG. 15A). Wgggen, et al., Evidence that nonsteroidal anti-inflammatory drugs decrease amyloid beta 42 production by direct modulation of gamma-secretase activity, J. Biol . Chem . 278: 3183-3187 (2003 Similar to)), Aβ42-inhibiting ginsenosides (eg, Rk1 and (20S) Rg3) are cell-free γ-secretases, although both compounds primarily affect Aβ42 production in cell-based assays. The test inhibited both Aβ40 and Aβ42 with similar efficacy (FIG. 15B).

Ginsenosides are metabolized by human enterobacteria after oral administration of ginseng extracts (Kobayashi K., et al., Metabolism of ginsenoside by human intestinal bacteria [II] Ginseng Review 1994; 18: 10-14; Hasegawa H., . et al, Main ginseng saponin metabolites formed by intestinal bacteria Planta Med 1996:.. 62: 453-457). For this reason, the effects on the Aβ42 production of two major metabolites of ginsenosides, including 20 (S) -protopanaxatriol (PPT) and 20 (S) -protopanaxadiol (PPD), were investigated. 20 (S) -Panaxtriol (PT) and 20 (S) -Panaxadiol (PD) are artificial derivatives of PPT and PPD, respectively. PPT or PT treatment reduced Aβ42 production without affecting the levels of Aβ42 in Neuro2a cells (Neuro2a-SW) expressing human Swedish variant forms of APP and CHO cells expressing wild type human APP (FIG. 16). PPD and PD did not give any inhibitory effect on Aβ40 or Aβ42 production.

In summary, Aβ42-inhibited natural compounds originating from heat-treated ginseng have been identified. Aβ42-inhibited ginsenosides, including Rk1 and (20S) Rg3, appear to specifically regulate γ-secretase activity involved in Aβ42 production. Structure-activity defines a class of compounds that can serve as the basis for the development of therapeutic agents that are effective in the treatment of AD.

Example  2

The benefits of ginsenoside therapy in the treatment of AD associated neurodegeneration can be demonstrated in the murine model of AD. Specifically, the ginsenoside compound (20S) Rg3, Rk1, Rg5, Rgk351 can be used to treat mice with AD-related neurodegeneration.

Mice expressing human APP and mice expressing Swedish familial Alzheimer's disease variant forms of APP can be purchased from Jackson Laboratory, 600 Main Street, Bar Harbor, Maine 04609. Thereafter, four groups of mice: (1) APP mice without ginsenoside treatment (placebo); (2) Swedish mice (placebo) without ginsenoside treatment; (3) APP mice + Rg5 (100 μg / μl / day); (4) Swedish mice + Rg5 (100 μg / μl / day) can be examined. After approximately 16 weeks of infusion therapy, the amount of Aβ 42 in the serum of mice can be measured. The results of this study are expected to demonstrate the overall benefits of ginsenoside therapy in the treatment of AD-related neurodegeneration. APP and Swedish mice without ginsenosides had significantly higher levels of serum Aβ42 and exhibited neurodegenerative behaviors compared to ginsenosides treated APP and Swedish mice.

Example  3

The true sagogenine of ginseng glycosides is structurally similar to some chemical components of other plants. Betufoliopolyenetriol (dammar-24-ene-3α, 12β, 20 (S) -triol) isolated from birch leaves is an intrinsic sapongenin of ginseng glycoside, 20 (S ) -Protopanaxadiol. For this reason, inexpensive and relatively readily available betulapolyentriol is suitable as a substrate for preparing 20 (S) -protopanaxadiol and its glycolsides Rg3, Rg5, Rk1.

Rkl or Rg5

Bettula poly entry is birch ( Btula) pendular ) leaves were extracted from the ether extract and then chromatographed on silica gel and crystallized from acetone: mp 195-195 °, lit. 197-198 ° (Fischer et al. (1959) Justus Liebigs Ann . Chem . 626: 185).

The 12-O-acetyl derivative of 20 (S) -protopanaxadiol (3) is prepared from betulapolyentriol in the series of reactions shown in Scheme 1. Betulapolyentriol is oxidized to ketone 1 [dammar-24-ene-12β, 20 (S) -diol-3-one: mp 197-199 °, lit 196-199 ° (yield: 60%)], It is acetylated with acetic anhydride dissolved in pyridine to form Compound 2 [12-O-Acetyl-dammar-24-ene-12β, 20 (S) -diol-3-one (yield: 100%?) (Nagal et. al., (1973) Chem. Pharm. Bull. 9: 2061). 1 H NMR (CDCl ₃ ) of Compound 2: 0.90 (s, 3H), 0.95 (s, 3H), 1.0 (s, 6H), 1.1 (s, 3H), 1.1 (s, 3H), 1.65 (s, 3H ), 1.72 (s, 3H), 2.1 (s, 3H), 3.04 (s, 1H), 4.73 (td, 1H), 5.17 (t, 1H). Sodium borohydride reduction of compound 2 in 2-propanol yielded compound 3 [12-O-Acetyl-dammar-24-ene-3β, 12β, 20 (S) -triol (yield: 90%)]. Form. 1 H NMR (CDCl ₃ ) of Compound 3: 0.78 (s, 3H), 0.86 (8, 3H), 0.95 (s, 3H), 1.0 (s, 3H), 1.02 (s, 3H), 1.13 (s, 3H ), 1.64 (s, 3H), 1.71 (s, 3H), 2.05 (s, 3H, OAc), 3.20 (dd, 1H, H-3α), 4.73 (td, 1H, H-12α), 5.16 (t , 1H, H-24).

Condensation of compound 3 with O-acetylate-sugar bromide in the presence of silver oxide and molecular sieve 4A in dichloroethane to form compound 4 (yield: 50%). Specifically, compound 3 (1.08 g, 2 mmol), silver oxide (1.4 g, 6 mmol), α-acetobromoglucose (2.47 g, 6 mmol), molecular sieve 4A (1.0 g), dichloroethane ( 20 ml) was stirred at room temperature until acetobromoglucose reacted (TLC). The reaction mixture was then diluted with CHCl ₃ and filtered. The solvent was evaporated and the residue was washed with hot water to remove excess glucose derivative. Silica gel column chromatography (8: 1 n-hexane-acetone) gave compound 4 (853 mg). Deprotection of the glucoside 4 affords ginsenoside Rg3, which is converted to Rkl or Rg5 in two steps.

Although the invention has been described in detail for clarity of understanding, various changes in form and detail are possible without departing from the precise scope of the invention in the appended claims.

Claims (51)

Compounds having the formula

R ₁ is selected from α-OH, β-OH, α-OX, β-OX, α-R ₆ COO-, β-R ₆ COO-, α-R ₆ PO _3- , β-R ₆ PO _3- Wherein X is a carbohydrate comprising one or more sugars or an acylated derivative thereof; R ₆ is alkenyl, aryl, or alkyl I; R ₂ is selected from H, OH, OAc, OX, wherein X is a carbohydrate comprising at least one sugar or an acylated derivative thereof; R ₃ is selected from H, OH, OAc; R ₄ is alkenyl, aryl, or alkyl II; R ₅ is H or OH. The compound of claim 1, wherein the alkyl I group further carries oxygen, nitrogen, or phosphorus. The compound of claim 1, wherein the alkyl II group further has a functional group selected from hydroxyl, ether, ketone, oxime, hydrazone, imine, and Schiff base. The compound of claim 1, wherein the sugar is selected from Glc, Ara (pyr), Ara (fur), Rha, Xyl. The compound of claim 1, wherein R ₄ is selected from:

Any stereo-center arrangement is R or S; X is OR or NR, wherein R is alkyl or aryl; X 'is alkyl, OR, NR, wherein R is alkyl or aryl; R 'is H, alkyl, or acyl. Use of a compound having the formula below in the treatment or prevention of pathological abnormalities:

R ₁ is selected from α-OH, β-OH, α-OX, β-OX, α-R ₆ COO-, β-R ₆ COO-, α-R ₆ PO _3- , β-R ₆ PO _3- Wherein X is a carbohydrate comprising one or more sugars or an acylated derivative thereof; R ₆ is alkenyl, aryl, or alkyl I; R ₂ is selected from H, OH, OAc, OX, wherein X is a carbohydrate comprising at least one sugar or an acylated derivative thereof; R ₃ is selected from H, OH, OAc; R ₄ is alkenyl, aryl, or alkyl II; R ₅ is H or OH. The compound of claim 6, wherein the alkyl I group further carries oxygen, nitrogen, or phosphorus; Alkyl II groups are characterized in that they further carry a functional group selected from hydroxyl, ether, ketone, oxime, hydrazone, imine, Schiff base. Use according to claim 6, characterized in that the pathological abnormality is neurodegeneration. Use according to claim 8, characterized in that the pathological abnormality is Alzheimer's disease. Use according to claim 6, characterized in that the pathological abnormality is an Aβ 42-associated disease. Isolated compounds having the formula:

R ₁ is selected from α-OH, β-OH, α-OX, β-OX, α-R ₆ COO-, β-R ₆ COO-, α-R ₆ PO _3- , β-R ₆ PO _3- Wherein X is a carbohydrate comprising one or more sugars or an acylated derivative thereof; R ₆ is alkenyl, aryl, or alkyl I; R ₂ is selected from H, OH, OAc, OX, wherein X is a carbohydrate comprising one or more sugars or acylated derivatives thereof; R ₃ is selected from H, OH, OAc; R ₄ is alkenyl, aryl, or alkyl II; R ₅ is H or OH. The compound of claim 10, wherein the alkyl I group further carries oxygen, nitrogen, or phosphorus; An alkyl II group is characterized in that it further has a functional group selected from hydroxyl, ether, ketone, oxime, hydrazone, imine, and Schiff base. A composition containing a compound having the formula:

R ₁ is selected from α-OH, β-OH, α-OX, β-OX, α-R ₆ COO-, β-R ₆ COO-, α-R ₆ PO _3- , β-R ₆ PO _3- Wherein X is a carbohydrate comprising one or more sugars or an acylated derivative thereof; R ₆ is alkenyl, aryl, or alkyl I; R ₂ is selected from H, OH, OAc, OX, wherein X is a carbohydrate comprising one or more sugars or acylated derivatives thereof; R ₃ is selected from H, OH, OAc; R ₄ is alkenyl, aryl, or alkyl II; R ₅ is H or OH. The compound of claim 13, wherein the alkyl I group further carries oxygen, nitrogen, or phosphorus; Alkyl II groups are characterized in that they further have a functional group selected from hydroxyl, ether, ketone, oxime, hydrazone, imine, Schiff base. Pharmaceutical compositions containing a compound having the formula: and a pharmaceutically acceptable carrier:

R ₁ is selected from α-OH, β-OH, α-OX, β-OX, α-R ₆ COO-, β-R ₆ COO-, α-R ₆ PO _3- , β-R ₆ PO _3- Wherein X is a carbohydrate comprising one or more sugars or an acylated derivative thereof; R ₆ is alkenyl, aryl, or alkyl I; R ₂ is selected from H, OH, OAc, OX, wherein X is a carbohydrate comprising at least one sugar or an acylated derivative thereof; R ₃ is selected from H, OH, OAc; R ₄ is alkenyl, aryl, or alkyl II; R ₅ is H or OH. The compound of claim 15, wherein the alkyl I group further carries oxygen, nitrogen, or phosphorus; Alkyl II group is a pharmaceutical composition, characterized in that it further has a functional group selected from hydroxyl, ether, ketone, oxime, hydrazone, imine, Schiff base. A method for synthesizing a compound having the formula below, comprising the following steps:

(a)

Treating a compound having the formula with an oxidizing agent to form a compound having the formula

(b) treating the compound formed in step (a) with a reducing agent to form a compound having the formula

R ₁ is H or OH; R ₂ is selected from H, OH, OAc, OX, wherein X is a carbohydrate comprising at least one sugar or an acylated derivative thereof; R ₃ is selected from H, OH, OAc; R ₄ is alkenyl, aryl, or alkyl. 18. The process according to claim 17, wherein the oxidant is chromic anhydride. 18. The method of claim 17, wherein the reducing agent is NaBH ₄ . 18. The method of claim 17, wherein a compound having the formula: is obtained from a plant:

The method of claim 20, wherein the plant is selected from conventional birch trees. The method of claim 20, wherein the compound having the formula is betulapolyenetriol.

A method for synthesizing a compound having the formula below, comprising the following steps:

(a)

Treatment of a compound having the formula with an oxidant to form a compound having the formula

(b) treating the compound formed in step (a) with a reducing agent to form a compound having the formula

(c) optionally, treating the compound formed in step (b) with a protected R ₁ derivative to form a compound having the formula

(d) The compound formed in step (c) is treated with a deprotection agent to form a compound having the formula

R ₁ is selected from α-OH, β-OH, α-OX, β-OX, α-R ₆ COO-, β-R ₆ COO-, α-R ₆ PO _3- , β-R ₆ PO _3- Wherein X is a carbohydrate comprising one or more sugars or an acylated derivative thereof; R ₆ is alkenyl, aryl, or alkyl I; R ₂ is selected from H, OH, OAc, OX, wherein X is a carbohydrate comprising at least one sugar or an acylated derivative thereof; R ₃ is selected from H, OH, OAc; R ₄ is alkenyl, aryl, or alkyl II; R ₅ is H or OH. The compound of claim 23, wherein the alkyl I group further carries oxygen, nitrogen, or phosphorus; The alkyl II group further comprises a functional group selected from hydroxyl, ether, ketone, oxime, hydrazone, imine, and Schiff base. The method of claim 23, wherein the oxidizing agent is chromic anhydride. The method of claim 23, wherein the reducing agent is NaBH ₄ . The method of claim 23, wherein the protected R ₁ derivative is a protected R ₁ halogen derivative. The method of claim 23, wherein the protected R ₁ derivative is protected by an Ac ₈ -group. The method of claim 28, wherein the compound is deprotected using NaOMe. The method of claim 23, wherein a compound having the formula: is obtained from a plant:

31. The method of claim 30, wherein the plant is selected from conventional birch trees. 31. The method of claim 30, wherein the compound having the formula: betulafolienetriol.

A method for synthesizing a compound having the formula below, comprising the following steps:

(a)

Treatment of a compound having the formula with an oxidant to form a compound having the formula

(b) treating the compound formed in step (a) with a protecting agent to form a compound having the formula

(c) treating the compound formed in step (b) with a reducing agent to form a compound having the formula

(d) treating the compound formed in step (c) with Ac ₈ -Glc-Glc-Br to form a compound having the formula

(e) treating the compound formed in step (d) with a deprotecting agent to form a compound having the formula

(f) further modifying the compound formed in step (e) to form a compound having the formula:

34. The method of claim 33, wherein the oxidant is chromic anhydride. The method of claim 33, wherein the reducing agent is NaBH ₄ . The method of claim 33, wherein the compound is deprotected using NaOMe. 34. The method of claim 33, wherein the compound having the formula: is obtained from a plant:

38. The method of claim 37, wherein the plant is selected from conventional birch trees. A method for synthesizing a compound having the formula below, comprising the following steps:

Compounds having the above formula are treated with reducing agents to form compounds having the following formula:

The method of claim 39, wherein the reducing agent is NaBH ₄ . A method for synthesizing a compound having the formula below, comprising the following steps:

(a)

Treatment of a compound having the above formula with a reducing agent forms a compound having the following formula:

(b) treating the compound formed in step (a) with Ac ₈ -Glc-Glc-Br to form a compound having the formula

(c) treating the compound formed in step (b) with a deprotecting agent to form a compound having the formula

42. The method of claim 41, wherein the reducing agent is NaBH ₄ . The method of claim 41, wherein the compound is deprotected using NaOMe. A method of treating or preventing pathological abnormalities in an individual, the method comprising administering to the individual a compound having the formula:

R ₁ is selected from α-OH, β-OH, α-OX, β-OX, α-R ₆ COO-, β-R ₆ COO-, α-R ₆ PO _3- , β-R ₆ PO _3- Wherein X is a carbohydrate comprising one or more sugars or an acylated derivative thereof; R ₆ is alkenyl, aryl, or alkyl I; R ₂ is selected from H, OH, OAc, OX, wherein X is a carbohydrate comprising at least one sugar or an acylated derivative thereof; R ₃ is selected from H, OH, OAc; R ₄ is alkenyl, aryl, or alkyl II; R ₅ is H or OH. The method of claim 44, wherein the alkyl I group further carries oxygen, nitrogen, or phosphorus; Alkyl II groups are characterized in that they further carry a functional group selected from hydroxyl, ether, ketone, oxime, hydrazone, imine, Schiff base. 45. The method of claim 44, wherein the pathological abnormality is neurodegeneration. 45. The method of claim 44, wherein the pathological abnormality is Alzheimer's disease. The method of claim 44, wherein the pathological abnormality is an Aβ 42 -associated disease. 45. The method of claim 44, wherein the subject is a human. A method of inhibiting β-amyloid production in a subject, the method comprising administering to the subject a compound having the formula:

R ₁ is selected from α-OH, β-OH, α-OX, β-OX, α-R ₆ COO-, β-R ₆ COO-, α-R ₆ PO _3- , β-R ₆ PO _3- Wherein X is a carbohydrate comprising one or more sugars or an acylated derivative thereof; R ₆ is alkenyl, aryl, or alkyl I; R ₂ is selected from H, OH, OAc, OX, wherein X is a carbohydrate comprising at least one sugar or an acylated derivative thereof; R ₃ is selected from H, OH, OAc; R ₄ is alkenyl, aryl, or alkyl II; R ₅ is H or OH. 51. The compound of claim 50, wherein the alkyl I group further carries oxygen, nitrogen, or phosphorus; Alkyl II groups are characterized in that they further carry a functional group selected from hydroxyl, ether, ketone, oxime, hydrazone, imine, Schiff base.

Images