KR20090087075A - Compounds useful for treating neurodegenerative disorders - Google Patents

Abstract

As described herein, the present invention provides compounds useful for treating or lessening the severity of a neurodegenerative disorder. The present invention also provides methods of treating or lessening the severity of such disorders wherein said method comprises administering to a patient a compound of the present invention, or composition thereof. Said method is useful for treating or lessening the severity of, for example, Alzheimer's disease. ® KIPO & WIPO 2009

Description

Compounds useful for treating neurodegenerative disorders

Cross Reference of Related Application

The present invention is a partial consecutive application of US patent application Ser. No. 11 / 434,726, filed May 16, 2006, which claims priority US Provisional Patent Application No. 60 / 681,662, filed May 17, 2005, each of which is incorporated herein by reference. Is incorporated herein by reference.

The present invention relates to pharmaceutically active compounds useful for treating or lessening the severity of neurodegenerative disorders.

An important role of long form amyloid beta-peptides, especially Aβ (1-42), in Alzheimer's disease has been established through various histopathology, genetics and biochemical studies. Selkoe, DJ, Physiol. Rev. 2001, 81: 741-766, Alzheimer's disease: genes, proteins, and therapy, and Younkin SG, J Physiol Paris. 1998, 92: 289-92, The role of A beta 42 in Alzheimer's disease. In particular, precipitation of Aβ (1-42) in the brain has been shown to be an early and constant characteristic of all forms of Alzheimer's disease. In fact, it occurs before the precipitation of Aβ (1-40), a shorter major form of A-beta, before the diagnosis of Alzheimer's disease is possible. Parvathy S, et al., Arch Neurol. 2001, 58: 2025-32, Correlation between Abetax-40-, Abetax-42-, and Abetax-43-containing amyloid plaques and cognitive decline]. A further relationship of Aβ (1-42) in disease etiology stems from the observation that mutations in the presenilin (gamma secretase) gene associated with the early onset of familial Alzheimer's disease consistently increase Aβ (1-42) levels. See Ishii K, et al., Neurosci Lett. 1997, 228: 17-20, Increased A beta 42 (43) -plaque deposition in early-onset familial Alzheimer's disease brains with the deletion of exon 9 and the missense point mutation (H163R) in the PS-1 gene]. Additional mutations in the amyloid precursor protein APP increase total Aβ and in some cases increase Aβ (1-42) only. Kosaka T, et al., Neurology, 48: 741-5, The beta APP717 Alzheimer mutation increases the percentage of plasma amyloid-beta protein ending at A beta42 (43)]. While various APP mutations can affect the form, quantity and location of precipitated Aβ, the predominant early species precipitated in the brain parenchyma has been found to be long Aβ (Mann). Mann DM, et al. , Am J Pathol. 1996, 148: 1257-66, Predominant deposition of amyloid-beta 42 (43) in plaques in cases of Alzheimer's disease and hereditary cerebral hemorrhage associated with mutations in amyloid precursor protein gene].

In the initial precipitation of Αβ, most precipitated proteins are in amorphous form or in the form of diffuse plaques, and substantially all of the Αβ is elongated. Gravina SA, et al., J Biol Chem, 270: 7013-6, Amyloid beta protein (A beta) in Alzheimer's disease brain. Biochemical and immunocytochemical analysis with antibodies specific for forms ending at A beta 40 or A beta 42 (43); Iwatsubo T, et al., Am J Pathol. 1996, 149: 1823-30, Full-length amyloid-beta (1-42 (43)) and amino-terminally modified and truncated amyloid-beta 42 (43) deposit in diffuse plaques; and Roher AE, et al., Proc Natl Acad Sci USA 1993, 90: 10836-40, beta-amyloid- (1-42) is a major component of cerebrovascular amyloid deposits: implications for the pathology of Alzheimer disease]. These initial precipitations of Aβ (1-42) can then make further precipitation of both long and short forms of Aβ (Tamaoka A, et al., Biochem Biophys Res Commun. 1994, 205: 834-42, Biochemical evidence for the long-tail form (A beta 1-42 / 43) of amyloid beta protein as a seed molecule in cerebral deposits of Alzheimer's disease].

In transgenic animals expressing A [beta], precipitation is associated with elevated levels of A [beta] (1-42), and the pattern of precipitation is a human body with A [beta] (1-42) precipitated after the initial A [beta] (1-40) precipitation. Similar to that observed in disease. Rockenstein E, et al., J Neurosci Res. 2001, 66: 573-82, Early formation of mature amyloid-beta protein deposits in a mutant APP transgenic model depends on levels of A beta (1-42); and Terai K, et al., Neuroscience 2001, 104: 299-310, beta-Amyloid deposits in transgenic mice expressing human beta-amyloid precursor protein have the same characteristics as those in Alzheimer's disease]. Similar patterns and settling times are observed in Down's syndrome patients with elevated Aβ expression and accelerated precipitation (Iwatsubo T, et al., Ann Neurol. 1995, 37: 294-9, Amyloid beta protein (A beta) deposition: A beta 42 (43) precedes A beta 40 in Down syndrome].

Thus, the selective degradation of Aβ (1-42) reduces neurodegeneration by reducing the likelihood of all forms of amyloid formation of Aβ, slowing or stopping the formation of new precipitates of Aβ, and inhibiting the formation of soluble toxic oligomers of Aβ. It appears as a disease-specific strategy that slows or stops the progression of the disorder.

The gist of the invention

As described herein, the present invention provides compounds useful for treating or lessening the severity of neurodegenerative disorders. The present invention also provides methods of treating or lessening the severity of such disorders, including administering a compound of the invention or a composition thereof to a patient. The method is useful, for example, in treating or lessening the severity of Alzheimer's disease.

1 shows the ¹ H NMR spectra of the chromatographic fractions sat14-9 and sat14-10.

2 shows the ¹ H NMR spectra of the chromatographic fractions sat14-11 and sat14-12.

3 shows the ¹ H NMR spectra of the chromatographic fractions sat15-1 and sat15-2.

4 shows the ¹ H NMR spectra of the chromatographic fractions sat15-4 and sat15-5.

FIG. 5 shows an enlarged view of C-18 reverse phase HPLC chromatogram separation of sat15-5, with numbers 1 to 5 corresponding to time windows for fractions sat16-1 to sat16-9.

6 shows the ¹ H NMR spectrum of fraction sat16-3 corresponding to compound 6 of purity 98%.

7 is a flow chart summary of separation protocol 2. FIG.

8 shows the HPLC traces of black cohosh extracts after semi-preparative HPLC.

9 shows the HPLC traces of compound 6 showing small deacyl peaks.

10 shows the HPLC trace amount of compound 6.

11 shows the mass spectrum of deacyl-compound 6.

12 shows the ¹ H NMR of deacyl-compound 6.

13 shows the HPLC trace amount of compound 6.

FIG. 14 shows ¹ H NMR (CD ₃ OD) of Compound 6.

15 shows the mass spectrum of compound 6.

FIG. 16 shows HPLC traces of Compound 6 detected at 205 nm isolated according to Protocol 2.

FIG. 17 shows HPLC traces of Compound 6 detected at 230 nm isolated according to Protocol 2.

FIG. 18 shows HPLC of Compound 6 detected in ELSD.

FIG. 19 shows a ¹ H NMR spectrum of Compound 6 isolated according to Protocol 2.

20 shows the mass spectrum of Compound 6 isolated according to Protocol 2. FIG.

21 shows the effect of IP-MS measurement of compound 6 on the relative amounts of amyloid-beta (1-40), (1-42), (1-37), (1-38) and (1-39) It is.

Figure 22 shows the IP- of compound 6 for the amounts of amyloid-beta (1-40), (1-42), (1-37), (1-38) and (1-39) of wild type and 717 mutant cells. MS measurement effect is shown.

Detailed Description of Certain Aspects of the Invention

1. General Description of Compounds of the Invention:

In one embodiment, the present invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof.

In Formula I above,

Each ring A, ring B, ring C, ring D and ring E are independently saturated or partially unsaturated aromatic rings,

G is S, CH ₂ , NR or O,

R ¹ and R ² are each independently halogen, R, OR, suitably protected hydroxyl group, SR, suitably protected thiol group, N (R) ₂ or suitably protected amino group, or R ¹ and R ² together form a 3 to 7 membered saturated or partially unsaturated aryl ring having 0 to 2 heteroatoms independently selected from nitrogen, oxygen and sulfur,

Each R is independently hydrogen, an optionally substituted C _1-6 aliphatic group, or an optionally substituted 3-8 membered saturated or partially unsaturated having 0-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur Is an aryl ring wherein two R on the same nitrogen atom are optionally substituted with 3 to 8 membered saturated or partially unsaturated aryl rings having 1 to 4 heteroatoms independently selected from nitrogen, oxygen and sulfur Form the

n is 0 to 2,

R ³ , R ⁴ , R ⁷ and R ⁸ are each independently halogen, R, OR, suitably protected hydroxyl group, SR, suitably protected thiol group, SO ₂ R, OSO ₂ R, N (R) ₂ , suitably protected amino group, NR (CO) R, NR (CO) (CO) R, NR (CO) N (R) ₂ , NR (CO) OR, (CO) OR, O (CO) R , (CO) N (R) ₂ and O (CO) N (R) ₂ ,

m is 0 to 2,

R ⁵ is TC (R ′) ₃ , TC (R ′) ₂ C (R ″) ₃ , R, OR, suitably protected hydroxyl group, SR, suitably protected thiol group, SO ₂ R, OSO ₂ R, N (R) ₂ , suitably protected amino group, NR (CO) R, NR (CO) (CO) R, NR (CO) N (R) ₂ , NR (CO) OR, (CO) OR , O (CO) R, (CO) N (R) ₂ or O (CO) N (R) _2, or

When R ⁵ is TC (R ') ₃ or TC (R') ₂ C (R ") ₃ , the R 'groups on R ⁵ and R ⁵ are optionally taken together, from 0 to 2 independently selected from nitrogen, oxygen and sulfur To form a 3-8 membered saturated or partially unsaturated aryl ring having 3 heteroatoms,

Each T is independently a valence bond, or an optionally substituted, linear or branched, saturated or unsaturated C _1-6 alkylidene chain, wherein up to two methylene units of T are optionally independently -O-,- Replaced with N (R)-, -S-, -C (O)-, -S (O)-or -S (O) _2- ,

Each R ′ and R ″ is independently R, OR, SR, SO ₂ R, OSO ₂ R, N (R) ₂ , NR (CO) R, NR (CO) (CO) R, NR (CO) N (R) ₂ , NR (CO) OR, (CO) OR, O (CO) R, (CO) N (R) ₂ and O (CO) N (R) ₂ ,

R ⁶ is halogen, R, OR, SR, SO ₂ R, OSO ₂ R, N (R) ₂ , NR (CO) R, NR (CO) (CO) R, NR (CO) N (R) ₂ , NR (CO) OR, (CO) OR, O (CO) R, (CO) N (R) ₂ or O (CO) N (R) ₂ ,

R ⁹ and R ^{9 '} are each independently selected from halogen, R, OR, SR and N (R) ₂ , or R ¹ and R ² together are 0 to 2 heteroatoms independently selected from nitrogen, oxygen and sulfur To form a 3-7 membered saturated or partially unsaturated aryl ring having

Q is a valence bond, or an optionally substituted linear or branched saturated or unsaturated C _1-6 alkylidene chain, wherein up to two methylene units of Q are optionally independently -O-, -N (R) -, -S-, -C (O)-, -S (O)-or -S (O) _2-

R ¹⁰ is R, suitably protected hydroxyl group, suitably protected thiol group, suitably protected amino group; Optionally substituted 3-8 membered saturated or partially unsaturated aryl monocyclic rings having 0-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur; Optionally substituted 8-10 membered saturated or partially unsaturated aryl bicyclic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur; Detectable residues, polymer residues, peptides or sugar-containing or sugar-like residues.

According to another embodiment, the present invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof.

Formula I

In Formula I above,

Each ring A, ring B, ring C, ring D and ring E are independently saturated or partially unsaturated aromatic rings,

G is S, CH ₂ , NR or O,

R ¹ and R ² are each independently halogen, R, OR, suitably protected hydroxyl group, SR, suitably protected thiol group, N (R) ₂ or suitably protected amino group, or R ¹ and R ² together form a 3 to 7 membered saturated or partially unsaturated aryl ring having 0 to 2 heteroatoms independently selected from nitrogen, oxygen and sulfur,

Each R is independently hydrogen, an optionally substituted C _1-6 aliphatic group, or an optionally substituted 3-8 membered saturated or partially unsaturated having 0-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur Is an aryl ring wherein two R on the same nitrogen atom are optionally substituted with 3 to 8 membered saturated or partially unsaturated aryl rings having 1 to 4 heteroatoms independently selected from nitrogen, oxygen and sulfur Form the

n is 0 to 2,

R ³ , R ⁴ , R ⁷ and R ⁸ are each independently halogen, R, OR, suitably protected hydroxyl group, SR, suitably protected thiol group, SO ₂ R, OSO ₂ R, N (R) ₂ , suitably protected amino group, NR (CO) R, NR (CO) (CO) R, NR (CO) N (R) ₂ , NR (CO) OR, (CO) OR, O (CO) R , (CO) N (R) ₂ and O (CO) N (R) ₂ ,

m is 0 to 2,

R ⁵ is TC (R ′) ₃ , TC (R ′) ₂ C (R ″) ₃ , R, OR, suitably protected hydroxyl group, SR, suitably protected thiol group, SO ₂ R, OSO ₂ R, N (R) ₂ , suitably protected amino group, NR (CO) R, NR (CO) (CO) R, NR (CO) N (R) ₂ , NR (CO) OR, (CO) OR , O (CO) R, (CO) N (R) ₂ or O (CO) N (R) _2, or R ⁵ is TC (R ') ₃ or TC (R') ₂ C (R ") ₃ In the case, the R 'group on R ⁶ and R ⁵ optionally together form a 3-8 membered saturated or partially unsaturated aryl ring having 0-2 heteroatoms independently selected from nitrogen, oxygen and sulfur,

Each T is independently a valence bond, or an optionally substituted, linear or branched, saturated or unsaturated C _1-6 alkylidene chain, wherein up to two methylene units of T are optionally independently -O-,- Replaced with N (R)-, -S-, -C (O)-, -S (O)-or -S (O) _2- ,

Each R ′ and R ″ is independently R, OR, SR, SO ₂ R, OSO ₂ R, N (R) ₂ , NR (CO) R, NR (CO) (CO) R, NR (CO) N (R) ₂ , NR (CO) OR, (CO) OR, O (CO) R, (CO) N (R) ₂ and O (CO) N (R) ₂ ,

R ⁶ is halogen, R, OR, SR, SO ₂ R, OSO ₂ R, N (R) ₂ , NR (CO) R, NR (CO) (CO) R, NR (CO) N (R) ₂ , NR (CO) OR, (CO) OR, O (CO) R, (CO) N (R) ₂ or O (CO) N (R) ₂ ,

R ⁹ and R ^{9 '} are each independently selected from halogen, R, OR, SR and N (R) ₂ , or R ¹ and R ² together are 0 to 2 heteroatoms independently selected from nitrogen, oxygen and sulfur To form a 3-7 membered saturated or partially unsaturated aryl ring having

Q is a valence bond, or an optionally substituted, linear or branched, saturated or unsaturated C _1-6 alkylidene chain, wherein up to two methylene units of Q are optionally independently -O-, -N (R) -, -S-, -C (O)-, -S (O)-or -S (O) _2-

R ¹⁰ is R, suitably protected hydroxyl group, suitably protected thiol group, suitably protected amino group; Optionally substituted 3-8 membered saturated or partially unsaturated aryl monocyclic rings having 0-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur; Optionally substituted 8-10 membered saturated or partially unsaturated aryl bicyclic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen and sulfur, detectable moieties, polymer residues, peptides or sugar-containing or Sugar-like residues,

only,

의 화합물은 제외된다.

The compound of is excluded.

2. Definition:

Compounds of the present invention include those described generally above, and are further illustrated by the embodiments, details, and types described herein. As used herein, the following definitions may apply unless stated otherwise. For the purposes of the present invention, chemical elements are identified according to the Periodic Table of the Elements (CAS version, Handbook of Chemistry and Physics, 75 ^th Ed). In addition, general principles of organic chemistry are described in "Organic Chemistry", Thomas Sorrell, University Science Books, Sausalito: 1999, and "March's Advanced Organic Chemistry", 5 ^th Ed., Ed .: Smith, MB and March , J., John Wiley & Sons, New York: 2001, the entirety of which is incorporated herein by reference.

As generally described above, ring A, ring B, ring C, ring D and ring E are each, independently, an aromatic ring that is saturated or partially unsaturated. It is understood that the compounds of the present invention may be considered as chemically viable compounds. Thus, those skilled in the art will understand that when any of ring A, ring B, ring C, ring D and ring E is unsaturated, certain substituents in that ring may not be present to satisfy the general rules of valence. For example, when ring D is unsaturated at the bond between ring D and ring E, R ⁶ may not be present. Alternatively, when ring D is unsaturated at the bond between ring D and ring C, R ⁸ and R ³ may not be present. All combinations of saturated and unsaturated ring A, ring B, ring C, ring D and ring E are contemplated by the present invention. Therefore, in order to satisfy the general rules of valence and depending on the saturation or unsaturation of ring A, ring B, ring C, ring D and ring E, R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , Presence or absence of each of R ⁷ , R ⁸ , R ⁹ , R ^{9 '} and QR ¹⁰ are considered accordingly.

As described herein, the compounds of the present invention may be optionally substituted with one or more substituents, for example, as generally described above or as exemplified in the specific classes, subclasses and types of the present invention. The phrase “optionally substituted” is understood to be used interchangeably with the phrase “substituted or unsubstituted”. In general, the term “substituted”, whether or not preceded by the term “optionally”, refers to the replacement of certain substituents of hydrogen radicals with radicals in a given structure. Unless stated otherwise, an optionally substituted group may have a substituent at each substitutable position of the group, and when one or more positions in a given structure may be substituted with one or more substituents selected from a particular group, the substituents may be It may be the same or different in position. Combinations of substituents contemplated by the present invention are preferably those which form stable or chemically viable compounds.

As used herein, the term “stable” refers to a compound that does not substantially change when placed under acceptable conditions for its preparation, detection, preferably its recovery, purification, and use for one or more purposes described herein. In some embodiments, a stable compound or chemically viable compound is a compound that does not substantially change when maintained at a temperature of 40 ° C. or less for at least one week in the absence of moisture or other chemical reaction conditions.

As used herein, the term "aliphatic" or "aliphatic group" is a straight (ie unbranched) or branched or unsubstituted hydrocarbon chain that is fully saturated or comprises one or more unsaturated units, or fully saturated or one or more unsaturated. By monocyclic hydrocarbons or bicyclic hydrocarbons is meant that is not aromatic (also referred to herein as “carbocycle”, “alicyclic” or “cycloalkyl”) that includes units but has a single point of attachment to the rest of the molecule. Unless stated otherwise, aliphatic groups contain 1-20 aliphatic carbon atoms. In some embodiments, aliphatic groups contain 1-6 aliphatic carbon atoms. In another embodiment, aliphatic groups contain 1-4 aliphatic carbon atoms. In certain embodiments, an “alicyclic” (or “carbocycle” or “cycloalkyl”) is completely saturated or comprises one or more unsaturated units, but in each bicyclic ring system any of the rings may contain from 3 to 7 members. By monocyclic C ₃ -C ₈ hydrocarbons or bicyclic C ₈ -C ₁₂ hydrocarbons which are not aromatic having a single point of attachment to the rest of the molecule having. Suitable aliphatic groups include, but are not limited to, linear or branched, substituted or unsubstituted alkyl, alkenyl, alkynyl groups and combination groups thereof, such as (cycloalkyl) alkyl, (cycloalkenyl) Alkyl or (cycloalkyl) alkenyl. In another embodiment, the aliphatic group is oxo (bivalent carbonyl oxygen atom = 0) or ring-forming substituents such as -O- (linear or branched alkylene or alkylidene) -O to form acetal or ketal It may have two geminal hydrogen atoms replaced with-.

In certain embodiments, exemplary aliphatic groups include, but are not limited to, ethynyl, 2-propynyl, 1-propenyl, 2-butenyl, 1,3-butadienyl, 2-pentenyl, vinyl Tenyl), allyl, isopropenyl, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, secondary-butyl, tert-butyl, pentyl, isopentyl, secondary-pentyl, neo-pentyl, tertiary- Pentyl, cyclopentyl, hexyl, isohexyl, secondary-hexyl, cyclohexyl, 2-methylpentyl, tert-hexyl, 2,3-dimethylbutyl, 3,3-dimethylbutyl, 1,3-dimethylbutyl and 2 , 3-dimethyl but-2-yl.

The terms "haloalkyl", "haloalkenyl" and "haloalkoxy" refer to alkyl, alkenyl or alkoxy and may be optionally substituted with one or more halogen atoms. The term "halogen" means F, Cl, Br or I. Such "haloalkyl", "haloalkenyl" and "haloalkoxy" groups may or may not be the same halogen and may have two or more halo substituents which may or may not be present at the same carbon atom. Examples include chloromethyl, periododomethyl, 3,3-dichloropropyl, 1,3-difluorobutyl, trifluoromethyl and 1-bromo-2-chloropropyl.

As used herein, the terms “heterocycle”, “heterocyclyl”, “heteroalicyclic” or “heterocyclic” refer to non-aromatic, monocyclic, bicyclic or in which one or more ring members are independently selected heteroatoms; Tricyclic ring system. In some embodiments, a “heterocycle”, “heterocyclyl”, “heteroalicyclic” or “heterocyclic” group is a heteroatom in which one or more ring members are independently selected from oxygen, sulfur, nitrogen and phosphorus, and in the system Each ring has 3 to 14 ring members, including 3 to 7 ring members.

The term "heteroatom" means one or more oxygen, sulfur, nitrogen, phosphorus or silicon [any oxidized form of nitrogen, sulfur, phosphorus or silicon; Quaternized forms of all basic nitrogen; Or a substitutable nitrogen of a heterocyclic ring, for example, N (in 3,4-dihydro-2H-pyrrolyl), NH (in pyrrolidinyl) or (in N-substituted pyrrolidinyl) NR ⁺ ].

As used herein, the term “unsaturated” means that the residue has one or more unsaturated units.

The term "alkoxy" or "thioalkyl" as used herein refers to an alkyl group as defined above attached to the main carbon chain via an oxygen ("alkoxy") or sulfur ("thioalkyl") atom.

The term "aryl" used alone or as part of a larger moiety in "aralkyl", "aralkoxy" or "aryloxyalkyl" means that at least one ring in the system is aromatic and each ring in the system has 3 to 7 Monocyclic, bicyclic, and tricyclic ring systems having a total of 5 to 14 ring members comprising two ring members. The term "aryl" can be used interchangeably with the term "aryl ring." The term "aryl" also refers to a heteroaryl ring system as described below.

The term “heteroaryl”, used alone or as part of a larger moiety in “heteroaralkyl” or “heteroarylalkoxy”, means that at least one ring in the system is aromatic and at least one ring in the system contains at least one heteroatom. , Monocyclic, bicyclic and tricyclic ring systems in which each ring in the system has a total of 5 to 14 ring members comprising 3 to 7 ring members. The term "heteroaryl" may be used interchangeably with the term "heteroaryl ring" or the term "heteroaromatic".

Aryl (including aralkyl, aralkoxy, aryloxyalkyl, etc.) or heteroaryl (including heteroaralkyl, heteroarylalkoxy, etc.) groups can include one or more substituents. Suitable substituents on the unsaturated carbon atoms of the aryl or heteroaryl group include halogen; N ₃ , CN, R °; OR °; SR °; 1,2-methylene-dioxy; 1,2-ethylenedioxy; Phenyl (Ph) optionally substituted with R °; -O (Ph) optionally substituted with R °; (CH ₂ ) _1-2 (Ph) optionally substituted with R °; CH═CH (Ph) optionally substituted with R °; NO ₂ ; CN; N (R °) ₂ ; NR ° C (O) R °; NR ° C (O) N (R °) ₂ ; NR ° CO ₂ R °; -NR ° NR ° C (O) R °; NR ° NR ° C (0) N (R °) ₂ ; NR ° NR ° CO ₂ R °; C (O) C (O) R °; C (O) CH ₂ C (O) R °; CO ₂ R °; C (O) R °; C (O) N (R °) ₂ ; 0C (0) N (R °) ₂ ; S (O) ₂ R °; SO ₂ N (R °) ₂ ; S (O) R °; NR ° SO ₂ N (R °) ₂ ; NR ° SO ₂ R °; C (= S) N (R °) ₂ ; C (= NH) -N (R °) ₂ ; And (CH ₂ ) _0-2 NHC (O) R °, wherein each independent presence of R ° is hydrogen, optionally substituted C _1-6 aliphatic, unsubstituted 5-6 membered heteroaryl or Selected from heterocyclic ring, phenyl, 0 (Ph) and CH ₂ (Ph), or despite the above definition, two independent presences of R ° in the same substituent or in different substituents, each R ° group is bonded Together with the atom (s) formed form a 3-8 membered cycloalkyl, heterocyclyl, aryl or heteroaryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen and sulfur. Optional substituents in the aliphatic group of R ° are N ₃ , CN, NH ₂ , NH (C _1-4 aliphatic), N (C _1-4 aliphatic) ₂ , halogen, C _1-4 aliphatic, OH, 0 (C _1-4 aliphatic), NO ₂ , CN, CO ₂ H, CO ₂ (C _1-4 aliphatic), O (haloC _1-4 aliphatic) and haloC _1-4 aliphatic group, wherein R ° Each of said C _1-4 aliphatic groups is unsubstituted.

Aliphatic or heteroaliphatic groups or nonaromatic heterocyclic rings may comprise one or more substituents. Suitable substituents on the saturated carbon of an aliphatic or heteroaliphatic group or non-aromatic heterocyclic ring are selected from those described above for the unsaturated carbon of the aryl or heteroaryl group, and further include 0 = 0, = S, = NNHR ^* , = NN ( R ^* ) ₂ , = NNHC (0) R ^* , = NNHCO ₂ (alkyl), = NNHSO ₂ (alkyl) or = NR ^* , wherein R ^* is each independently hydrogen and an optionally substituted C _1-6 Selected from aliphatic groups. Any substituent in the aliphatic group of R ^* is NH ₂ , NH (C _1-4 aliphatic), N (C _1-4 aliphatic) ₂ , halogen, C _1-4 aliphatic, OH, 0 (C _1-4 aliphatic) , NO ₂ , CN, CO ₂ H, CO ₂ (C _1-4 aliphatic), O (halo C _1-4 aliphatic) and halo (C _1-4 aliphatic), wherein R ^* is C _1- Each of the _four aliphatic groups is unsubstituted.

Optional substituents on the nitrogen of the non-aromatic heterocyclic ring are R ⁺ , N (R ⁺ ) ₂ , C (O) R ⁺ , CO ₂ R ⁺ , C (O) C (O) R ⁺ , C (O) CH ₂ C (O) R ⁺ , SO ₂ R ⁺ , SO ₂ N (R ⁺ ) ₂ , C (= S) N (R ⁺ ) ₂ , C (= NH) -N (R ⁺ ) ₂ and NR ⁺ SO ₂ is selected from R ⁺ , wherein R ⁺ is hydrogen, optionally substituted C _1-6 aliphatic, optionally substituted phenyl, optionally substituted O (Ph), optionally substituted CH ₂ (Ph), optionally substituted (CH ₂ ) _1-2 (Ph); Optionally substituted CH═CH (Ph); Or an unsubstituted 5- to 6-membered heteroaryl or heterocyclic ring having 1 to 4 heteroatoms independently selected from oxygen, nitrogen and sulfur, or despite the definitions above, R in the same substituent or in a different substituent two independent presence of ^+, each R ⁺ group is bound atomic nitrogen with (s), oxygen, and 3-to 8 circle having the 0 to 4 heteroatoms independently selected from sulfur cycloalkyl, hetero To form a cyclyl, aryl or heteroaryl ring. Any substituent in the aliphatic group or phenyl ring of R ⁺ is NH ₂ , NH (C _1-4 aliphatic), N (C _1-4 aliphatic) ₂ , halogen, C _1-4 aliphatic, OH, 0 (C _{1- 4} aliphatic), NO ₂ , CN, CO ₂ H, CO ₂ (C _1-4 aliphatic), O (halo C _1-4 aliphatic) and halo (C _1-4 aliphatic), wherein R ⁺ Each C _1-4 aliphatic group is unsubstituted.

의 2개의 존재로 치환될 때, 이들 R°의 2개의 존재는 이들이 결합된 산소 원자와 함께 융합된 6원 산소 함유 환

을 형성한다). 다양한 다른 환은 R°(또는 R⁺ 또는 본원에 유사하게 정의된 다른 변수)의 2개의 독립적인 존재가 각각의 변수가 결합된 원자(들)과 함께 형성될 수 있고, 상기 기재된 예들은 이에 제한되고자 하는 것이 아닌 것으로 이해되어야 한다.As described above, in certain embodiments, two independent presences of R ° (or R ⁺ or other variables similarly defined herein) may be derived from nitrogen, oxygen, and sulfur together with the atom (s) to which each variable is bound. To form a 3-8 membered cycloalkyl, heterocyclyl, aryl or heteroaryl ring having 0-4 heteroatoms independently selected. Exemplary two rings of R ° (or R ⁺ or other variables similarly defined herein) include, but are not limited to, the following examples where each variable is formed with the atom (s) to which it is bound A) two independent presences of R ° (or R ⁺ or other variables similarly defined herein), which are bonded to the same atom and form a ring with the atom, for example N (R °) ₂ Wherein both the presence of R ° together with the nitrogen atom form a piperidin-1-yl, piperazin-1-yl or morpholin-4-yl group; And b) two independent presences of R ° (or R ⁺ or other variables similarly defined herein) which are bonded to different atoms and together with both atoms form a ring, for example (where a phenyl group Is OR °

When substituted with two beings, the two beings of these R ° are 6-membered oxygen containing rings fused together with the oxygen atoms to which they are attached

To form). Various other rings may be formed with two independent presence of R ° (or R ⁺ or other variables similarly defined herein) with the atom (s) to which each variable is bound, the examples described above being intended to be limited thereto. It should be understood that it is not.

As used herein, the term “detectable moiety” is used interchangeably with the term “label” to mean all moieties that can be detected, eg, a first label and a second label. First labels, such as radioisotopes (eg ³² P, ³³ P, ³⁵ S or ¹⁴ C), mass-tags and fluorescent labels can be detected without further alteration of the reporter reporter group to be.

As used herein, the term "second label" refers to biotin and various protein antigens that require the presence of a residue, eg, a second intermediate, for the production of a detectable signal. In the case of biotin, the second intermediate may comprise a streptavidin-enzyme conjugate. For antigen labels, the second intermediate may comprise an antibody-enzyme conjugate. Some fluorescent groups function as second labels as they transfer energy to another group in the process of non-radiative fluorescence resonance energy transfer (FRET), and the second group produces a detected signal.

As used herein, the terms “fluorescent label”, “fluorescent dye” and “fluorophore” refer to moieties that absorb light energy at defined excitation wavelengths and emit light energy at different wavelengths. Examples of fluorescent labels include, but are not limited to, Alexa Fluor dyes (Alexa Fluor 350, Alexa Fluor 488, Alexa Fluor 532, Alexa Fluor 546, Alexa Fluor 568, Alexa Fluor 594, Alexa Fluor 633, Alexa Fluor 660 and Alexa Fluor 680), AMCA, AMCA-S, BODIPY dyes (Bodypie FL, Bodypie R6G, Bodypie TMR, Bodypie TR, Bodypie 530/550, Bodypie 558/568, Bodypie 564/570, Bodhi 576/589, Bodhi 581/591, Bodhi 630/650 and Bodhi 650/665), Carboxyrodamine 6G, Carboxy-X-Rhodamine (ROX), Cascade Blue , Cascade Yellow, Coumarin 343, Cyanine Dyes (Cy3, Cy5, Cy3.5 and Cy5.5), Monosil, Dapoxyl, Dialkylaminocoumarin, 4 ', 5'-dichloro -2 ', 7'-dimethoxy-fluorescein, DM-NERF, Eosin, erythrosin, fluorescein, FAM, hydroxyku Lynn, IRDyes (IRD 40, IRD 700 and IRD 800), JOE, Lysamine Rhodamine B, Marina Blue, Methoxycoumarin, Naphthofluorescein, Oregon Green 488, Oregon Green 500 , Oregon Green 514, Pacific Blue, PyMPO, Pyrene, Rhodamine B, Rhodamine 6G, Rhodamine Green, Rhodamine Red, Rhodol Green, 2 ', 4', 5 ', 7'-Tetra-Bro Parent sulfone-fluorescein, tetramethyl-rhodamine (TMR), carboxytetramethylhodamine (TAMRA), Texas Red, Texas Red-X.

As used herein, the term "mass-tag" refers to all residues that can be detected solely by their mass using mass spectroscopy (MS) detection techniques. Examples of mass-tags are electrophoretic release tags, such as N- [3- [4 '-[(p-methoxytetrafluorobenzyl) oxy] phenyl] -3-methylglyceronyl] isonipcoat Acids, 4 '-[2,3,5,6-tetrafluoro-4- (pentafluorophenoxyl)] methyl acetophenone and derivatives thereof. Synthesis and use of these mass-tags are described in US Patent 4,650,750, US Patent 4,709,016, US Patent 5,360,8191, US Patent 5,516,931, US Patent 5,602,273, US Patent 5,604,104, US Patent No. 5,610,020 and US Pat. No. 5,650,270. Other examples of mass-tags include, but are not limited to, nucleotides, dideoxynucleotides, oligonucleotides of various lengths and base compositions, oligopeptides, oligosaccharides, and other synthetic polymers of various lengths and monomer compositions. Large variety of organic molecules, neutral and charged (biomolecules or synthetic compounds) in suitable mass ranges (100-2,000 daltons) may also be used as mass-tags.

As used herein, the term “substrate” means any substance or macromolecular complex to which the functionalized end-groups of the compounds of the invention may be attached. Examples of commonly used substrates include, but are not limited to, glass surfaces, silica surfaces, plastic surfaces, metal surfaces, metal or chemical coating-containing surfaces, membranes (such as nylon, polysulfone or silica), micro-beads (Such as latex, polystyrene or other polymers), porous polymer matrices (such as polyacrylamide gels, polysaccharides or polymethacrylates), macromolecular complexes (such as proteins or polysaccharides).

Unless stated otherwise, the structures shown herein also include all isomeric forms of the structure, such as enantiomers, diastereomers, and geometric isomers (or form isomers); For example, R and S configurations for each asymmetric center, (Z) and (E) double bond isomers and (Z) and (E) shape isomers are meant. Thus, single stereochemical isomers and mixtures of enantiomers, diastereomers and geometric isomers (or form isomers) of the compounds of the invention are within the scope of the invention.

Unless stated otherwise, all tautomeric forms of the compounds of the invention are within the scope of the invention.

Additionally, unless stated otherwise, the structures shown herein are also meant to include different compounds only in the presence of one or more isotope-rich atoms. For example, compounds having the structure of the present invention are within the scope of the present invention, except for replacing hydrogen with deuterium or tritium or replacing carbon with ¹³ C-rich or ¹⁴ C-rich carbon. Such compounds are useful, for example, as analytical tools or probes in biological assays.

3. Description of Exemplary Compounds:

As generally described above, the G moiety of formula I is S, CH ₂ , NR or O. In certain embodiments, the G residue of Formula (I) is O.

As generally described above, R ¹ and R ² of formula I are each independently halogen, R, OR, suitably protected hydroxyl group, SR, suitably protected thiol group, N (R) ₂ or suitably Or a protected amino group, or R ¹ and R ² together form a 3-7 membered saturated or partially unsaturated aryl ring having 0-2 heteroatoms independently selected from nitrogen, oxygen, and sulfur. In certain embodiments, R ¹ and R ² of Formula I are each independently R or OR. In another embodiment, R ¹ and R ² of Formula I are each independently R, wherein R is hydrogen or an optionally substituted C _1-6 aliphatic group. According to another aspect of the invention, R ¹ and R ² of formula I together are a 3-6 membered saturated or partially unsaturated having 0 to 2 heteroatoms independently selected from nitrogen, oxygen and sulfur To form an aryl ring. Another aspect of the invention provides compounds of formula I, wherein R ¹ and R ² together form a 3- to 6-membered saturated carbocyclic ring. In another embodiment, R ¹ and R ² of Formula I together form a cyclopropyl ring.

In certain embodiments, the n residue of formula (I) is 0 or 1. In another embodiment, the n residue of Formula I is 1.

As generally described above, the R ⁵ groups of formula I are selected from TC (R ′) ₃ , TC (R ′) ₂ C (R ″) ₃ , R, OR, suitably protected hydroxyl groups, SR, suitably Protected thiol group, SO ₂ R, OSO ₂ R, N (R) ₂ , suitably protected amino group, NR (CO) R, NR (CO) (CO) R, NR (CO) N (R) ₂ , NR (CO) OR, (CO) OR, 0 (CO) R, (CO) N (R) ₂ or 0 (CO) N (R) ₂ , wherein each T is independently a valence bond, or optionally As a substituted linear or branched saturated or unsaturated C _1-6 alkylidene chain, up to two methylene units of T are optionally independently -O-, -N (R)-, -S-, -C ( Is replaced by O) —, —S (O) — or —S (O) ₂ — In certain embodiments, each T is independently a valence bond, or a straight or branched C _1-4 alkylidene chain, wherein T One methylene unit of is optionally replaced with -0-, -N (R)-or -S- In other embodiments, each T is independently a valence bond, or a linear or branched C _1-4 egg Kildene chains In another embodiment, each T is a valence bond.

When the R ⁵ group of Formula I is TC (R ') ₃ or TC (R') ₂ C (R ") ₃ , R 'and R" are each independently R, OR, SR, SO ₂ R, OSO ₂ R, N (R) ₂ , NR (CO) R, NR (CO) (CO) R, NR (CO) N (R) ₂ , NR (CO) OR, (CO) OR, 0 (CO) R, And (CO) N (R) ₂ and 0 (CO) N (R) ₂ . In certain embodiments, R 'and R "are each independently R, OR, OC (O) R, SR, or N (R) _{2. In} another embodiment, R' and R" are each independently R, OR, or OC (O) R. Exemplary R ′ and R ″ groups include hydrogen, CH ₃ , OH and OC (O) CH ₃ .

In general, as described above, when R ⁵ is TC (R ′) ₃ or T—CH (R ′) C (R ″) ₃ , the R ′ groups on R ⁶ and R ⁵ are optionally together, nitrogen, oxygen and Form a 3-8 membered saturated or partially unsaturated aryl ring having 0-2 heteroatoms independently selected from sulfur In certain embodiments, R ⁵ is TC (R ′) ³ or TC (R ′ ) ₂ C (R ") ₃ , and the R 'group on R ⁶ and R ⁵ together form a 5-7 membered saturated ring having 1 or 2 heteroatoms independently selected from nitrogen, oxygen and sulfur. Form. In another embodiment, R ⁵ is TC (R ′) ₃ or TC (R ′) ₂ C (R ″) ₃ and the R ′ groups on R ⁶ and R ⁵ together are a 6 membered saturated member having 1 oxygen atom. When T is a valence bond, the compound is a compound of Formula (IIa) when R ⁵ is TC (R ') ₃ , wherein the compound is R ⁵ is TC (R') ₂ C (R " ₃ ) is a compound of Formula IIb:

In the above formulas IIa and IIb,

Each R ', R ", R ¹ , R ² , R ³ , R ⁴ , R ⁷ , R ⁸ , R ⁹ , R ^9' , Q and R ¹⁰ are generally classified above and as described above and herein and As described in the subdivisions

As described generally above, the R ⁵ group of formula (I) is, by itself, a suitably protected hydroxyl group, suitably protected thiol group or suitably protected amino group. Hydroxyl protecting groups are well known in the art and described by reference: Protecting Groups in Organic Synthesis, TW Greene and PGM Wuts, 3 rd edition, John Wiley & Sons, 1999, its specialty of which are incorporated herein by reference; It includes those described in detail. Examples of suitably protected hydroxyl groups of the R ⁵ group of formula (I) further include, but are not limited to, esters, allyl ethers, ethers, silyl ethers, alkyl ethers, arylalkyl ethers and alkoxyalkyl ethers. . Examples of such esters include formate, acetate, carbonate and sulfonate. Specific examples include formate, benzoyl formate, chloroacetate, trifluoroacetate, methoxyacetate, triphenylmethoxyacetate, p-chlorophenoxyacetate, 3-phenylpropionate, 4-oxopentanoate, 4 , 4- (ethylenedithio) pentanoate, pivaloate (trimethylacetyl), crotonate, 4-methoxy-crotonate, benzoate, p-benylbenzoate, 2,4,6-trimethylbenzo Ates, carbonates such as methyl, 9-fluorenylmethyl, ethyl, 2,2,2-trichloroethyl, 2- (trimethylsilyl) ethyl, 2- (phenylsulfonyl) ethyl, vinyl, allyl and p-nitrobenzyl. Examples of such silyl ethers include trimethylsilyl, triethylsilyl, t-butyldimethylsilyl, t-butyldiphenylsilyl, triisopropylsilyl and other trialkylsilyl ethers. Alkyl ethers include methyl, benzyl, p-methoxybenzyl, 3,4-dimethoxybenzyl, trityl, t-butyl, allyl and allyloxycarbonyl ethers or derivatives. Alkoxyalkyl ethers include acetals such as methoxymethyl, methylthiomethyl, (2-methoxyethoxy) methyl, benzyloxymethyl, beta- (trimethylsilyl) ethoxymethyl and tetrahydropyranyl ether . Examples of arylalkyl ethers include benzyl, p-methoxybenzyl (MPM), 3,4-dimethoxybenzyl, 0-nitrobenzyl, p-nitrobenzyl, p-halbenzyl, 2,6-dichlorobenzyl, p-sia Nobenzyl, 2-picolinyl and 4-picolinyl.

Thiol protecting groups are well known in the art literature, in the Reference Protecting Groups in Organic Synthesis, TW Greene and PGM Wuts, 3 rd edition, John Wiley & Sons, 1999, its specialty of which are incorporated herein by reference; Includes those described in detail. Suitably protected thiol groups of the R ⁵ residue of formula (I) include, but are not limited to, disulfides, thioethers, silyl thioethers, thioesters, thiocarbonates, thiocarbamates, and the like. Examples of this group include, but are not limited to, but not limited to, alkyl thioethers, benzyl and substituted benzyl thioethers, triphenylmethyl thioethers, trichloroethoxycarbonyl.

According to another aspect of the invention, the R ⁵ residue of formula (I) is a thiol protecting group removable under neutral conditions, for example AgNO ₃ , HgCl ₂ and the like. Other neutral conditions include reduction with a suitable reducing agent. Suitable reducing agents are dithiothreitol (DTT), mercaptoethanol, dithionite, reduced glutathione, reduced glutathridin, reduced thioredoxin, substituted phosphines, such as triscarboxyethyl phosphine ( TCEP) and all other peptides or organic reducing agents or other reagents known to those skilled in the art. According to another aspect of the invention, the R ⁵ residue of formula (I) is a "photodegradable" thiol protecting group. Such suitable thiol protecting groups are known in the art and include, but are not limited to, nitrobenzyl groups, tetrahydropyranyl (THP) groups, trityl groups, -CH ₂ SCH ₃ (MTM), dimethylmethoxymethyl or -CH ₂ -SS-pyridin- ₂ -yl. Those skilled in the art will appreciate that many suitable hydroxyl protecting groups described herein are also suitable as thiol protecting groups.

In certain embodiments, the R ⁵ groups of Formula I are suitably protected amino groups. Amino protecting groups are well known in the art and described in Protecting Groups in Organic Synthesis, TW Greene and PGM Wuts, 3 ^rd edition, John Wiley & Sons, 1999, the entirety of which is incorporated herein by reference. Includes those described in detail. Suitably protected amino groups of these R ⁵ residues further include, but are not limited to, aralkylamines, carbamates, cyclic imides, allyl amines, amides, and the like. Examples of this group include t-butyloxycarbonyl (BOC), ethyloxycarbonyl, methyloxycarbonyl, trichloroethyloxycarbonyl, allyloxycarbonyl (Alloc), benzyloxocarbonyl (CBZ), allyl, Phthalimide, benzyl (Bn), fluorenylmethylcarbonyl (Fmoc), formyl, acetyl, chloroacetyl, dichloroacetyl, trichloroacetyl, phenylacetyl, trifluoroacetyl, benzoyl and the like. In certain embodiments, the amino protecting group of the R ⁵ residue is phthalimido. In another embodiment, the amino protecting group of the R ⁵ residue is a tert-butyloxycarbonyl (BOC) group.

As generally described above, the Q group of formula (I) is a valence bond, or an optionally substituted linear or branched, saturated or unsaturated C _1-4 alkylidene chain, wherein up to two methylene units of Q are optionally Are independently replaced with -O-, -N (R)-, -S-, -C (O)-, -S (O)-or -S (O) _2- . In certain embodiments, Q is an optionally substituted linear or branched saturated or unsaturated C _1-2 alkylidene chain, wherein one or less methylene units of Q are optionally -O-, -N (R)-or Replaced by -S- In other embodiments, Q is -O-.

In general, as described above, the R ¹⁰ groups of formula (I) are selected from the group consisting of R, suitably protected hydroxyl group, suitably protected thiol group, suitably protected amino group; Optionally substituted 3-8 membered saturated or partially unsaturated aryl monocyclic rings having 0-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur; An optionally substituted 8-10 membered saturated or partially unsaturated aryl bicyclic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen and sulfur, a detectable moiety, a polymer residue, a peptide or a sugar- Containing groups or sugar-like groups.

이다. 또 다른 양태에 따르면, 화학식 I의 R¹⁰ 그룹은

이다. 또 다른 양태는 R¹⁰이

인 화학식 I의 화합물을 제공한다.In certain embodiments, the R ¹⁰ groups of formula I are sugar-containing groups. Such sugar-containing groups are well known to those skilled in the art and described in "Essential of Glycobiology" Edited by Varki, A., et al., Cold Spring Harbor Laboratory Press. Cold Spring Harbor, NY 2002. In certain embodiments, the R ¹⁰ group of formula I is glycoside. Exemplary R ¹⁰ groups include arabinopyranosides and xylopyranosides. In certain embodiments, the R ¹⁰ group of formula (I) is xylopyranoside. In certain embodiments, the R ¹⁰ group of formula (I) is arabinofyranoside. In another embodiment, the R ¹⁰ groups of formula I are

to be. According to another embodiment, the R ¹⁰ groups of formula (I)

to be. Another embodiment is that R ¹⁰ is

It provides a compound of formula (I).

According to another aspect of the invention, the R ¹⁰ group of formula I is a sugar-like. Such sugar-analogs are well known to those skilled in the art and include those described in detail in "Essential of Glycobiology". For example, sugar-like groups contemplated in the present invention include cyclitol and the like. In certain embodiments, R ¹⁰ is a cyclitol moiety, wherein the cyclitol is a cycloalkane containing one hydroxyl group in each of at least three ring atoms as defined in the IUPAC Treaty. In other embodiments, the cyclitol moiety comprises inositol, such as scyllo-inositol.

In addition, suitable sugar-like residues of the R ¹⁰ group of formula (I) include acyclic sugar groups. This group includes only a few examples of linear alkitol and erythritol. It is understood that sugar groups can exist in cyclic or acyclic forms. Thus, acyclic forms of sugar groups are contemplated by the present invention as suitable sugar-like residues of the R ¹⁰ groups of formula (I).

In certain embodiments, the R ¹⁰ groups of formula I are detectable moieties. In another embodiment, the R ¹⁰ groups of formula (I) are fluorescent labels, fluorescent dyes or fluorophores as defined above.

According to another aspect of the invention, the R ¹⁰ group of formula (I) is a polymer moiety. Polymer residues are well known in the art and described in "Chemistry of Protein Conjugation and Cross-Linking" Shan S. Wong, CRC Press. Boca Raton, Florida. 1991]. Suitable polymer moieties of the R ¹⁰ group of formula (I) include poly (alkylene oxides) such as PEG, poly (amino acids) and other polymer moieties that can be conjugated to the compounds of the invention.

As generally described above, the R ¹⁰ group of formula (I) is, by itself, a suitably protected hydroxyl group, suitably protected thiol group or suitably protected amino group. Hydroxyl protecting groups are well known in the art and described by reference: Protecting Groups in Organic Synthesis, TW Greene and PGM Wuts, 3 rd edition, John Wiley & Sons, 1999, its specialty of which are incorporated herein by reference; It includes those described in detail. Examples of suitable hydroxyl protecting groups of the R ¹⁰ group of formula (I) further include, but are not limited to, esters, allyl ethers, ethers, silyl ethers, alkyl ethers, arylalkyl ethers and alkoxyalkyl ethers. Examples of such esters include formate, acetate, carbonate and sulfonate. Specific examples include formate, benzoyl formate, chloroacetate, trifluoroacetate, methoxyacetate, triphenylmethoxyacetate, p-chlorophenoxyacetate, 3-phenylpropionate, 4-oxopentanoate, 4 , 4- (ethylenedithio) pentanoate, pivaloate (trimethylacetyl), crotonate, 4-methoxy-crotonate, benzoate, p-benylbenzoate, 2,4,6-trimethylbenzo Ates, carbonates such as methyl, 9-fluorenylmethyl, ethyl, 2,2,2-trichloroethyl, 2- (trimethylsilyl) ethyl, 2- (phenylsulfonyl) ethyl, vinyl, allyl and p-nitrobenzyl. Examples of such silyl ethers include trimethylsilyl, triethylsilyl, t-butyldimethylsilyl, t-butyldiphenylsilyl, triisopropylsilyl and other trialkylsilyl ethers. Alkyl ethers include methyl, benzyl, p-methoxybenzyl, 3,4-dimethoxybenzyl, trityl, t-butyl, allyl and allyloxycarbonyl ethers or derivatives. Alkoxyalkyl ethers include acetals such as methoxymethyl, methylthiomethyl, (2-methoxyethoxy) methyl, benzyloxymethyl, beta- (trimethylsilyl) ethoxymethyl and tetrahydropyranyl ether . Examples of arylalkyl ethers are benzyl, p-methoxybenzyl (MPM), 3,4-dimethoxybenzyl, O-nitrobenzyl, p-nitrobenzyl, p-halobenzyl, 2,6-dichlorobenzyl, p-sia Nobenzyl, 2-picolinyl and 4-picolinyl.

Thiol protecting groups are well known in the art literature, in the Reference Protecting Groups in Organic Synthesis, TW Greene and PGM Wuts, 3 rd edition, John Wiley & Sons, 1999, its specialty of which are incorporated herein by reference; Includes those described in detail. Suitable thiol protecting groups of the R ¹⁰ residue of formula (I) include, but are not limited to, disulfides, thioethers, silyl thioethers, thioesters, thiocarbonates, thiocarbamates and the like. Examples of this group include, but are not limited to, but not limited to, alkyl thioethers, benzyl and substituted benzyl thioethers, triphenylmethyl thioethers, trichloroethoxycarbonyl.

According to another aspect of the invention, the R ¹⁰ residue of formula (I) is a thiol protecting group which is removable under neutral conditions, for example AgNO ₃ , HgCl ₂ and the like. Other neutral conditions include reduction with a suitable reducing agent. Suitable reducing agents are dithiothreitol (DTT), mercaptoethanol, dithionite, reduced glutathione, reduced glutathridin, reduced thioredoxin, substituted phosphines, such as triscarboxyethyl phosphine ( TCEP) and all other peptides or organic reducing agents or other reagents known to those skilled in the art. According to another aspect of the invention, the R ¹⁰ residue of formula (I) is a "photodegradable" thiol protecting group. Such suitable thiol protecting groups are known in the art and include, but are not limited to, nitrobenzyl groups, tetrahydropyranyl (THP) groups, trityl groups, -CH ₂ SCH ₃ (MTM), dimethylmethoxymethyl or -CH ₂ -SS-pyridin- ₂ -yl. Those skilled in the art will appreciate that many suitable hydroxyl protecting groups described herein are also suitable as thiol protecting groups.

In certain embodiments, the R ¹⁰ groups of formula I are suitably protected amino groups. Amino protecting groups are well known in the art literature, in the Reference Protecting Groups in Organic Synthesis, TW Greene and PGM Wuts, 3 rd edition, John Wiley & Sons, 1999, its specialty of which are incorporated herein by reference; Includes those described in detail. Suitable amino protecting groups of these R ¹⁰ residues further include, but are not limited to, aralkylamines, carbamates, cyclic imides, allyl amines, amides, and the like. Examples of this group include t-butyloxycarbonyl (BOC), ethyloxycarbonyl, methyloxycarbonyl, trichloroethyloxycarbonyl, allyloxycarbonyl (Alloc), benzyloxocarbonyl (CBZ), allyl, Phthalimide, benzyl (Bn), fluorenylmethylcarbonyl (Fmoc), formyl, acetyl, chloroacetyl, dichloroacetyl, trichloroacetyl, phenylacetyl, trifluoroacetyl, benzoyl and the like. In certain embodiments, the amino protecting group of the R ¹⁰ residue is phthalimido. In another embodiment, the amino protecting group of the R ¹⁰ residue is a tert-butyloxycarbonyl (BOC) group.

In certain embodiments, the present invention provides a compound of formula I comprising each stereoisomer, wherein the compound is

의 화합물 중의 1개, 2개 또는 3개 모두는 제외된다.

One, two or all three of the compounds of are excluded.

As generally described above, the present invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof:

Formula I

In Formula I above,

Each of the variables is as described above and in the classifications and subclasses described above and herein.

In certain embodiments, the present invention provides a compound of Formula (I) or a pharmaceutically acceptable salt thereof having the stereochemistry as shown in Formula (Ia):

In Formula Ia above,

Each of the variables is as described above for the compounds of Formula I and in the classifications and subclasses described above and herein.

In certain embodiments, the R ¹ and R ² groups of formula I together form a 3-7 membered saturated or partially unsaturated aryl ring having 0-2 heteroatoms independently selected from nitrogen, oxygen, and sulfur do. In another embodiment, the R ¹ and R ² groups of formula I together form a 3 to 6 membered saturated ring having 0 to 2 heteroatoms independently selected from nitrogen, oxygen and sulfur. In another embodiment, the R ¹ and R ² groups of formula I together form a 3-6 membered saturated carbocyclic ring. According to another aspect of the invention there is provided a compound of formula (Ib) or a pharmaceutically acceptable salt thereof:

In Formula Ib above,

Each of the variables is as described above for the compounds of Formula I and in the classifications and subclasses described above and herein.

In another embodiment, the present invention provides a compound of Formula (Ic) or a pharmaceutically acceptable salt thereof:

In Formula Ic above,

Each of the variables is as described above for the compounds of Formula I and in the classifications and subclasses described above and herein.

As generally described above, ring A, ring B, ring C, ring D and ring E are each, independently, an aromatic ring that is saturated or partially unsaturated. In certain embodiments, ring B is unsaturated and R ¹ and R ² are absent so that a compound of Formula II or a pharmaceutically acceptable salt thereof is formed:

In Formula II above,

Each of the variables is as described above for the compounds of Formula I and in the classifications and subclasses described above and herein.

In certain embodiments, the n group of formula II is 0 or 1 and the G group of formula II is oxygen.

According to another aspect, the present invention provides a compound of Formula (IIa) or a pharmaceutically acceptable salt thereof:

Formula IIa

In Formula IIa above,

Each of the variables is as described above for the compounds of Formula I and in the classifications and subclasses described above and herein.

In certain embodiments, the n group of formula IIa is 0 or 1 and the G group of formula IIa is oxygen.

In another embodiment, Ring B and Ring D are both unsaturated and R ¹ , R ² and R ⁶ are absent, thus forming a compound of Formula III or a pharmaceutically acceptable salt thereof:

In Formula III above,

Each of the variables is as described above for the compounds of Formula I and in the classifications and subclasses described above and herein.

In certain embodiments, the n group of formula III is 0 or 1 and the G group of formula III is oxygen.

According to another embodiment, the present invention provides a compound of Formula IV or a pharmaceutically acceptable salt thereof:

In Formula IV above,

Each of the variables is as described above for the compounds of Formula I and in the classifications and subclasses described above and herein.

는 단일 또는 이중 결합을 나타낸다. 당업자는

가 이중 결합을 나타낼 때, R⁶이 존재하지 않는다는 것을 이해할 것이다. 반대로,

가 단일 결합일 때, R⁶은 존재한다. 따라서, 특정 양태에서,

는 이중 결합을 나타내고, R⁶은 존재하지 않는다. 다른 양태에서,

는 단일 결합을 나타내고, R⁶은 상기 정의된 바와 같다.As used herein

Represents a single or double bond. Those skilled in the art

When will represent a double bond, it will be understood that R ⁶ is absent. Contrary,

When is a single bond, R ⁶ is present. Thus, in certain embodiments,

Represents a double bond, and R ⁶ is absent. In another aspect,

Represents a single bond and R ⁶ is as defined above.

According to another aspect, the present invention provides a compound of Formula IVa or a pharmaceutically acceptable salt thereof. "

In Formula IVa above,

Each of the variables is as described above for the compounds of Formula I and in the classifications and subclasses described above and herein.

In certain embodiments, the G group of formula IVa is oxygen. In other embodiments, the R ⁴ groups of Formula IVa are R, OR or suitably protected hydroxyl groups. In another embodiment, the R ⁴ group of formula IVa is R.

Another aspect of the invention relates to a compound of formula IVb or a pharmaceutically acceptable salt thereof:

In Formula IVb above,

Each of the variables is as described above for the compounds of Formula I and in the classifications and subclasses described above and herein.

In certain embodiments, the R ¹ and R ² groups of formula IVb together form a 3-7 membered saturated or partially unsaturated aryl ring having 0-2 heteroatoms independently selected from nitrogen, oxygen, and sulfur. do. In another embodiment, the R ¹ and R ² groups of formula IVb together form a 3 to 6 membered saturated ring having 0 to 2 heteroatoms independently selected from nitrogen, oxygen and sulfur. In another embodiment, the R ¹ and R ² groups of formula IVb together form a three to six membered saturated carbocyclic ring. According to another aspect of the present invention there is provided a compound of Formula IVc or a pharmaceutically acceptable salt thereof:

In Formula IVc above,

Each of the variables is as described above for the compounds of Formula I and in the classifications and subclasses described above and herein.

In certain embodiments, the R ⁷ group of formula IVc is -OH.

According to another aspect of the present invention there is provided a compound of Formula IVd or a pharmaceutically acceptable salt thereof:

In Formula IVd above,

Each of the variables is as described above for the compounds of Formula I and in the classifications and subclasses described above and herein.

In certain embodiments, the R ⁷ group of formula IVd is -OH.

In another embodiment, the present invention provides a compound of Formula V or a pharmaceutically acceptable salt thereof:

In Formula V above,

Each ring A, ring B, ring C, ring D and ring E are independently saturated or partially unsaturated aromatic rings,

G is S, CH ₂ , NR or O,

R ¹ and R ² are each independently halogen, R, OR, suitably protected hydroxyl group, SR, suitably protected thiol group, N (R) ₂ or suitably protected amino group, or R ¹ and R ² together form a 3 to 7 membered saturated or partially unsaturated aryl ring having 0 to 2 heteroatoms independently selected from nitrogen, oxygen and sulfur,

Each R is independently hydrogen, an optionally substituted C _1-6 aliphatic group, or an optionally substituted 3-8 membered saturated or partially unsaturated having 0-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur Is an aryl ring wherein two R on the same nitrogen atom are optionally substituted with 3 to 8 membered saturated or partially unsaturated aryl rings having 1 to 4 heteroatoms independently selected from nitrogen, oxygen and sulfur Form the

n is 0 to 2,

R ³ , R ⁴ , R ⁷ and R ⁸ are each independently halogen, R, OR, suitably protected hydroxyl group, SR, suitably protected thiol group, SO ₂ R, OSO ₂ R, N (R) ₂ , suitably protected amino group, NR (CO) R, NR (CO) (CO) R, NR (CO) N (R) ₂ , NR (CO) OR, (CO) OR, O (CO) R , (CO) N (R) ₂ and O (CO) N (R) ₂ ,

m is 0 to 2,

R ⁵ is TC (R ′) ₃ , TC (R ′) ₂ C (R ″) ₃ , R, OR, suitably protected hydroxyl group, SR, suitably protected thiol group, SO ₂ R, OSO ₂ R, N (R) ₂ , suitably protected amino group, NR (CO) R, NR (CO) (CO) R, NR (CO) N (R) ₂ , NR (CO) OR, (CO) OR , O (CO) R, (CO) N (R) ₂ or O (CO) N (R) ₂ ,

Each T is independently a valence bond, or an optionally substituted linear or branched, saturated or unsaturated C _1-6 alkylidene chain, wherein up to two methylene units of T are optionally independently -O-,- Replaced by N (R)-, -S-, -C (O)-, -S (O)-or -S (O) _2-

Each R ′ and R ″ is independently R, OR, SR, SO ₂ R, OSO ₂ R, N (R) ₂ , NR (CO) R, NR (CO) (CO) R, NR (CO) N (R) ₂ , NR (CO) OR, (CO) OR, O (CO) R, (CO) N (R) ₂ and O (CO) N (R) ₂ ,

R ⁶ is halogen, R, OR, SR, SO ₂ R, OSO ₂ R, N (R) ₂ , NR (CO) R, NR (CO) (CO) R, NR (CO) N (R) ₂ , NR (CO) OR, (CO) OR, O (CO) R, (CO) N (R) ₂ or O (CO) N (R) ₂ ,

R ⁹ and R ^{9 '} are each independently selected from halogen, R, OR, SR and N (R) ₂ , or R ¹ and R ² together are 0 to 2 heteroatoms independently selected from nitrogen, oxygen and sulfur To form a 3-7 membered saturated or partially unsaturated aryl ring having

Q is a valence bond, or an optionally substituted, linear or branched, saturated or unsaturated C _1-6 alkylidene chain, wherein up to two methylene units of Q are optionally independently -O-, -N (R) Is replaced by-, -S-, -C (O)-, -S (O)-or -S (O) _2- .

The embodiments described herein for compounds of formula (I) apply to each variable of the compound of formula (V), alone or in combination.

In certain embodiments, Q is -O-. In other embodiments, Q is -NH-.

In another aspect, the present invention,

(A) providing a compound of formula (Vb) or a pharmaceutically acceptable salt thereof; and

There is provided a process for preparing a compound of formula Va or a pharmaceutically acceptable salt thereof, comprising the step (b) of treating a compound of formula Vb with a suitable enzyme to form a compound of formula Va:

In the above formulas Va and Vb,

Each ring A, ring B, ring C, ring D and ring E are independently saturated or partially unsaturated aromatic rings,

G is S, CH ₂ , NR or O,

R ¹ and R ² are each independently halogen, R, OR, suitably protected hydroxyl group, SR, suitably protected thiol group, N (R) ₂ or suitably protected amino group, or R ¹ and R ² together form a 3 to 7 membered saturated or partially unsaturated aryl ring having 0 to 2 heteroatoms independently selected from nitrogen, oxygen and sulfur,

Each R is independently hydrogen, an optionally substituted C _1-6 aliphatic group, or an optionally substituted 3-8 membered saturated or partially unsaturated having 0-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur Is an aryl ring wherein two R on the same nitrogen atom are optionally substituted with 3 to 8 membered saturated or partially unsaturated aryl rings having 1 to 4 heteroatoms independently selected from nitrogen, oxygen and sulfur Form the

n is 0 to 2,

R ³ , R ⁴ , R ⁷ and R ⁸ are each independently halogen, R, OR, suitably protected hydroxyl group, SR, suitably protected thiol group, SO ₂ R, OSO ₂ R, N (R) ₂ , suitably protected amino group, NR (CO) R, NR (CO) (CO) R, NR (CO) N (R) ₂ , NR (CO) OR, (CO) OR, O (CO) R , (CO) N (R) ₂ and O (CO) N (R) ₂ ,

m is 0 to 2,

R ⁵ is TC (R ′) ₃ , TC (R ′) ₂ C (R ″) ₃ , R, OR, suitably protected hydroxyl group, SR, suitably protected thiol group, SO ₂ R, OSO ₂ R, N (R) ₂ , suitably protected amino group, NR (CO) R, NR (CO) (CO) R, NR (CO) N (R) ₂ , NR (CO) OR, (CO) OR , O (CO) R, (CO) N (R) ₂ or O (CO) N (R) ₂ ,

Each T is independently a valence bond, or an optionally substituted linear or branched, saturated or unsaturated C _1-6 alkylidene chain, wherein up to two methylene units of T are optionally independently -O-,- Replaced by N (R)-, -S-, -C (O)-, -S (O)-or -S (O) _2-

Each R ′ and R ″ is independently R, OR, SR, SO ₂ R, OSO ₂ R, N (R) ₂ , NR (CO) R, NR (CO) (CO) R, NR (CO) N (R) ₂ , NR (CO) OR, (CO) OR, O (CO) R, (CO) N (R) ₂ or O (CO) N (R) ₂ ,

R ⁶ is halogen, R, OR, SR, SO ₂ R, OSO ₂ R, N (R) ₂ , NR (CO) R, NR (CO) (CO) R, NR (CO) N (R) ₂ , NR (CO) OR, (CO) OR, O (CO) R, (CO) N (R) ₂ or O (CO) N (R) ₂ ,

R ⁹ and R ^{9 '} are each independently selected from halogen, R, OR, SR and N (R) ₂ , or R ¹ and R ² together are 0 to 2 heteroatoms independently selected from nitrogen, oxygen and sulfur To form a 3-7 membered saturated or partially unsaturated aryl ring having

R ¹⁰ is a sugar-containing or sugar-like residue.

The embodiments described herein apply to the variables of the compounds of the formulas Va and Vb alone or in combination.

As used herein, the term “suitable enzyme” refers to any enzyme capable of removing the R ¹⁰ sugar residues of the compound of formula Vb to form a compound of formula Va, ie, hydrolyzing glycosidic bonds. Such enzymes are known to those skilled in the art. In certain embodiments, suitable enzymes are cellulase, xylanase, xylosidase, glycyrrhizin hydrolase or glucuronidase. In certain embodiments, a suitable enzyme is cellulase. In another embodiment, the sugar moiety of the R ¹⁰ group of formula Vb is arabinofyranoside or xylopyranoside. Details of this modification are described in the Examples section below.

In certain embodiments, the present invention provides a compound of Formula Vc, Vd, Ve, Vf, Vg or Vh or a pharmaceutically acceptable salt thereof:

In the above formulas Vc, Vd, Ve, Vf, Vg and Vh,

Each variable is as defined for a compound of Formula (I) herein.

According to another aspect, the present invention provides a compound of Formula VI or a pharmaceutically acceptable salt thereof:

In Formula VI above,

Each ring A, ring B, ring C, ring D and ring E are independently saturated or partially unsaturated aromatic rings,

G is S, CH ₂ , NR or O,

R ¹ and R ² are each independently halogen, R, OR, suitably protected hydroxyl group, SR, suitably protected thiol group, N (R) ₂ or suitably protected amino group, or R ¹ and R ² together form a 3 to 7 membered saturated or partially unsaturated aryl ring having 0 to 2 heteroatoms independently selected from nitrogen, oxygen and sulfur,

Each R is independently hydrogen, an optionally substituted C _1-6 aliphatic group, or an optionally substituted 3-8 membered saturated or partially unsaturated having 0-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur Is an aryl ring wherein two R on the same nitrogen atom are optionally substituted with 3 to 8 membered saturated or partially unsaturated aryl rings having 1 to 4 heteroatoms independently selected from nitrogen, oxygen and sulfur Form the

R ³ , R ⁴ , R ⁷ and R ⁸ are each independently halogen, R, OR, suitably protected hydroxyl group, SR, suitably protected thiol group, SO ₂ R, OSO ₂ R, N (R) ₂ , suitably protected amino group, NR (CO) R, NR (CO) (CO) R, NR (CO) N (R) ₂ , NR (CO) OR, (CO) OR, O (CO) R , (CO) N (R) ₂ and O (CO) N (R) ₂ ,

R ⁶ is halogen, R, OR, SR, SO ₂ R, OSO ₂ R, N (R) ₂ , NR (CO) R, NR (CO) (CO) R, NR (CO) N (R) ₂ , NR (CO) OR, (CO) OR, O (CO) R, (CO) N (R) ₂ or O (CO) N (R) ₂ ,

R ⁹ and R ^{9 '} are each independently selected from halogen, R, OR, SR and N (R) ₂ , or R ¹ and R ² together are 0 to 2 heteroatoms independently selected from nitrogen, oxygen and sulfur To form a 3-7 membered saturated or partially unsaturated aryl ring having

Q is a valence bond, or an optionally substituted, linear or branched, saturated or unsaturated C _1-6 alkylidene chain, wherein up to two methylene units of Q are optionally independently -O-, -N (R) -, -S-, -C (O)-, -S (O)-or -S (O) _2-

R ¹⁰ is R, suitably protected hydroxyl group, suitably protected thiol group, suitably protected amino group; Optionally substituted 3-8 membered saturated or partially unsaturated aryl monocyclic rings having 0-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur; Optionally substituted 8-10 membered saturated or partially unsaturated aryl bicyclic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen and sulfur, detectable moieties, polymer residues, peptides, or sugar-containing Or sugar-like residues.

Embodiments described herein with respect to compounds of Formula (I) apply alone or in combination to compounds of Formula (VI).

In another aspect, the present invention,

Providing a compound of formula VIa or a salt thereof (a) and

Provided is a process for preparing a compound of formula VI or a pharmaceutically acceptable salt thereof, comprising the step (b) of removing the acetyl group of formula VIa to form a compound of formula VI:

Formula VI

In Formulas VI and VIa above,

Each ring A, ring B, ring C, ring D and ring E are independently saturated or partially unsaturated aromatic rings,

G is S, CH ₂ , NR or O,

R ¹ and R ² are each independently halogen, R, OR, suitably protected hydroxyl group, SR, suitably protected thiol group, N (R) ₂ or suitably protected amino group, or R ¹ and R ² together form a 3 to 7 membered saturated or partially unsaturated aryl ring having 0 to 2 heteroatoms independently selected from nitrogen, oxygen and sulfur,

Each R is independently hydrogen, an optionally substituted C _1-6 aliphatic group, or an optionally substituted 3-8 membered saturated or partially unsaturated having 0-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur Is an aryl ring wherein two R on the same nitrogen atom are optionally substituted with 3 to 8 membered saturated or partially unsaturated aryl having 1 to 4 heteroatoms independently selected from nitrogen, oxygen and sulfur, together with said nitrogen atom; Forming a ring,

R ³ , R ⁴ , R ⁷ and R ⁸ are each independently halogen, R, OR, suitably protected hydroxyl group, SR, suitably protected thiol group, SO ₂ R, OSO ₂ R, N (R) ₂ , suitably protected amino group, NR (CO) R, NR (CO) (CO) R, NR (CO) N (R) ₂ , NR (CO) OR, (CO) OR, O (CO) R , (CO) N (R) ₂ and O (CO) N (R) ₂ ,

R ⁶ is halogen, R, OR, SR, SO ₂ R, OSO ₂ R, N (R) ₂ , NR (CO) R, NR (CO) (CO) R, NR (CO) N (R) ₂ , NR (CO) OR, (CO) OR, O (CO) R, (CO) N (R) ₂ or O (CO) N (R) ₂ ,

R ⁹ and R ^{9 '} are each independently selected from halogen, R, OR, SR and N (R) ₂ , or R ¹ and R ² together are 0 to 2 heteroatoms independently selected from nitrogen, oxygen and sulfur To form a 3-7 membered saturated or partially unsaturated aryl ring having

Q is a valence bond, or an optionally substituted, linear or branched, saturated or unsaturated C _1-6 alkylidene chain, wherein up to two methylene units of Q are optionally independently -O-, -N (R) -, -S-, -C (O)-, -S (O)-or -S (O) _2-

R ¹⁰ is R, suitably protected hydroxyl group, suitably protected thiol group, suitably protected amino group; Optionally substituted 3-8 membered saturated or partially unsaturated aryl monocyclic rings having 0-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur; Optionally substituted 8-10 membered saturated or partially unsaturated aryl bicyclic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen and sulfur, detectable moieties, polymer residues, peptides, or sugar-containing Or sugar-like residues.

Embodiments described herein with respect to compounds of Formula (I) apply alone or in combination to compounds of Formula (VIa).

In certain embodiments, removal of the acetyl group in step (b) is achieved by treatment with a suitable base. In another embodiment, removal of the acetyl group in step (b) is achieved by heating in a suitable solvent. In another embodiment, removal of the acetyl group in step (b) is achieved by heating at about 40-115 ° C. In another embodiment, step (b) is performed at about 60 to about 90 ° C.

Suitable solvents are single solvents or solvent mixtures that can facilitate the reaction process. Suitable solvents may dissolve one or more reaction components, or alternatively, suitable solvents may facilitate stirring of the suspension of one or more reaction components. Examples of suitable solvents useful in the present invention are protic solvents, halogenated hydrocarbons, ethers, esters, aromatic hydrocarbons, polar or nonpolar aprotic solvents or any mixture thereof. Such mixtures may be, for example, mixtures of protic and aprotic solvents such as benzene / methanol / water; Benzene / water; DME / water and the like. In certain embodiments, suitable solvents are polar aprotic solvents such as alcohols.

Suitable bases are reagents that are sufficiently basic to remove the acetyl group of the compound of formula VIa to form a compound of formula VI. In certain embodiments, the suitable base is a suitable solvent itself. As an example thereof, it has been found that the methanolic solution of the compound of formula VIa is sufficiently basic to remove acetyl groups to form the compound of formula VI. Details of the modifications are described in the Examples section below. In other embodiments, a suitable solvent is an organic base. In another embodiment, a suitable base is an alkali or alkaline earth alkoxide or hydroxide. Such bases include sodium methoxide or sodium hydroxide. In another embodiment, a suitable base is ammonia.

In certain embodiments, the present invention provides a compound of Formula VII or VIII or a pharmaceutically acceptable salt thereof:

In the above formulas VII and VIII,

Each variable is as defined herein for a compound of Formula (I).

In another embodiment, the present invention provides a compound of Formula IX or X or a pharmaceutically acceptable salt thereof:

In the above formulas IX and X,

Each variable is as defined herein for a compound of Formula (I).

Exemplary compounds of the invention are shown in Table 1 below:

TABLE 1

Exemplary compounds of Formulas IVa and Vg are shown in Table 2 below:

TABLE 2

4. General Methods for Providing Compounds of the Invention:

Compounds of the present invention can generally be prepared or isolated by methods of synthesis and / or semi-synthesis known to those skilled in the art for similar compounds and by the methods detailed in the Examples below.

Isolation of the Active Ingredient

Specific compounds of the invention when Simi fugue La Simulation (cimicifuga racemosa) or sub-La Simulation also separated from the known black cohosh root sh (actaea racemosa) in akta and structure of these compounds was found. Commercial extracts, powders and capsules of black cohosh roots can be used to treat a variety of postmenopausal and gynecological disorders. Surprisingly, however, certain compounds present in black cohosh roots have been found to be useful for modulating and / or inhibiting amyloid-beta peptide production. In particular, certain compounds have been isolated and identified from black cohosh roots, where these compounds are useful for modulating and / or inhibiting amyloid-beta peptides, in particular amyloid-beta peptides (1-42) production. The compound is included by formula (I). These compounds can generally be isolated and used in a form that is substantially free of other compounds present in the roots. Alternatively, extracts can be prepared from the roots, which extracts are abundant in the compounds of the present invention.

Some compounds of the present invention, as described above and herein, are isolated from standard extracts of cultured or wild black cohosh roots and rhizomes. It is also contemplated that the compounds of the present invention may be isolated from plant root tissue or culture medium of culture plant tissue grown in culture. This method of growing plant root tissue in culture is well known to those skilled in the art and is described in Hairy Roots, Cultrue and Application, edited by Pauline M. Doran, published by Harwood Academic Publishers, Amsterdam, The Netherlands. Copyright 1997 OPA (Overseas Publishers Association) Amsterdam B.V. ISBN 90-5702-117-X, the entirety of which is incorporated herein by reference.

Alternatively, the compounds of the present invention can be prepared by semisynthetic processes starting from black cohosh and other compounds found in related extracts of the class of shimishipu, regardless of the root and root or root of the plant. Although it is not a skilled synthetic precursor in the art, and thus limited, Fushimi when fugue La Simulation, Fushimi when fugue dahu Rica (Cimicifuga dahurica), Fushimi when fugue included in the peptidase (Cimicifuga foetida), Fushimi when fugue spatula clay polyamic (Cimicifuga heracleifolia), Simi during Fugue japonica (Cimicifuga japonica), Simi during Fugue Oh Serena (Cimicifuga acerina), Simi during Fugue acetic Lima (Cimicifuga acerima), Simi during Fugue simplex (Cimicifuga simplex), Simi during Fugue Ella other (Cimicifuga elata ), Fushimi when fugue polyamic (Cimicifuga calthaefolia) in kalta, Fushimi when fugue free fall (Cimicifuga frigida), Fushimi when fugue la shinny Atta (Cimicifuga laciniata), Fushimi when fugue Mai ray Simi when fugue Ruby (Cimicifuga mairei), polyamic ( Cimicifuga rubifolia), during Simi Fugue Americana (Cimicifuga americana), a non-hotel Simi during fugue is displayed (Cimicifuga biternata) and Fugue in BP during SIMI It will be appreciated that one or more of the siminifupu , including Cimicifuga bifida , may be obtained from a species or variant thereof. This can be accomplished by chemical or biological modification of the isolated compound or extract fraction or mixture of compounds. Chemical modifications can be accomplished by manipulation of temperature, pH and / or treatment with various solvents, without limitation. Biological modifications can be accomplished by treatment of isolated compounds or extract fractions or mixtures of compounds with plant tissues, plant tissue extracts, other microbial organisms or isolated enzymes from any organism.

In certain embodiments, the present invention provides an extract of black cohosh root, wherein the extract comprises at least 10% by weight of a compound of the present invention. In another embodiment, the present invention provides an extract of black cohosh root, wherein the extract comprises about 10% to about 50% by weight of the compound of the present invention. In another embodiment, the present invention provides an extract of black cohosh root, the extract comprising about 10% to about 50% by weight of the compound of the present invention, wherein the extract is substantially free of actin.

According to another embodiment, the present invention provides a compound of formula (I) which is substantially free of other compounds found in black cohosh roots. The term "substantially free" as used herein means that the compound consists of a significantly higher proportion of the compound of formula (I) compared to the compound found in the black cohosh root or extract thereof. In some embodiments, the present invention provides from about 1% to about 99% by weight of the compound of formula (I). In certain embodiments, the compound of formula I is provided in greater than about 80% chemical purity. In another embodiment, the compound of formula (I) is provided in greater than about 90% chemical purity. In another embodiment, the compound of formula (I) comprises up to about 10.0 area% HPLC of other components of the black cohosh root with respect to the total area of the HPLC chromatogram. In another embodiment, the compound of formula (I) comprises no greater than about 8.0 area% HPLC of other components of the black cohosh root, and in other embodiments no greater than about 3 area%, relative to the total area of the HPLC chromatogram.

Methods of determining whether a compound of the present invention is in a form that is substantially free of other compounds commonly found in black cohosh roots are known to those skilled in the art as described below. Compounds previously isolated and identified from black cohosh roots include certain cycloartha including acteols, acetylactenols, 26-deoxyactenols, simigogenols, actins, 26-deoxyactins and simimifugosides. Nol triterpene is included. (E) -isoferulic acid and isoflavone formmononetin have also been isolated and identified. Representative these compounds have the structure:

Accordingly, another embodiment of the present invention is formula I substantially free of one or more acteols, acetylactenos, 26-deoxyactenos, simigogenols, actins, 26-deoxyactins and simimifugosides It provides a compound of. In certain embodiments, the present invention provides a compound of Formula I substantially free of acteols, acetylactenos, 26-deoxyactenos, simigogenols, actins, 26-deoxyactins and simimifugosides do.

According to another embodiment, the present invention provides formula I having reduced amounts of one or more acteols, acetylactenos, 26-deoxyactenos, simigogenols, actins, 26-deoxyactins and simimifugosides Provides extracts of black cohosh roots rich in compounds. According to another embodiment, the present invention provides compounds of formula I having reduced amounts of acteols, acetylactenos, 26-deoxyactenols, simigogenols, actins, 26-deoxyactins and simimifugosides, respectively. Provides extracts of black cohosh roots rich in compound.

Various techniques for the extraction, separation and / or purification of each active ingredient of black cohosh roots are well known in the art. The present invention encompasses both the identification of such active ingredients as described herein and the incorporation of such ingredients into the compositions of the present invention as described herein.

Each active ingredient of the black cohosh extract can be identified as described herein and can be isolated and / or purified using techniques known in the art. The active ingredient may be purified from the root itself in other forms or above all by decoction of an extract of the invention or a mixture of commercially available extracts. Various techniques available for purification include filtration, selective precipitation, extraction with organic solvents, extraction with aqueous solvents, column chromatography (silica gel), high performance liquid chromatography (HPLC) and other methods known to those skilled in the art. .

In certain embodiments, the extract of the present invention uses fractions isolated from black cohosh roots. By separated fraction is meant an auxiliary amount of root material which is removed, for example, by chromatographic means, distillation, precipitation, extraction, filtration or by other means from the root itself. In other embodiments, the root extracts and fractions are removed from them by chromatography, distillation, precipitation or extraction. Such extraction and separation techniques are well known to those skilled in the art. Details of some of these techniques are described in the Examples section below.

According to another aspect of the present invention, the presence and purity of the active compound are only a few examples including nuclear magnetic spectrometer (NMR), mass spectrometer, infrared spectrometer (IR), ultraviolet visible light spectrometer, elemental analysis and polarimeter, refractometer Evaluate by chemical method. Such assays are known to those skilled in the art. In other embodiments, the chemical structures of the active compounds isolated from the black cohosh roots are only a few, including, for example, NMR, mass spectrometers, infrared spectrometers (IR), ultraviolet visible light spectrometers, elemental analysis and polarimeters, refractometers and X-ray crystallography. It measures by the method known to those skilled in the art including.

Although some exemplary embodiments have been described above and herein, it is understood that the root extracts of the present invention may be prepared according to the methods generally described above using appropriate starting materials by methods generally available to those skilled in the art.

5. Uses, Formulations and Administration

Pharmaceutically Acceptable Compositions

According to another aspect of the present invention, pharmaceutically acceptable compositions are provided, wherein these compositions comprise all of the compounds described herein and optionally comprise a pharmaceutically acceptable carrier, adjuvant or vehicle. In certain embodiments, the composition optionally further comprises one or more additional therapeutic agents.

It is also understood that certain compounds of the present invention may exist in free form for treatment or, where appropriate, as pharmaceutically acceptable salts thereof.

As used herein, the term “pharmaceutically acceptable salts” is within the scope of sound medical judgment and suitable for use in contact with tissues of humans and lower animals without undue toxicity, irritation, allergic reactions, etc., and a reasonable benefit / harm ratio. Means a salt to be compensated for. A “pharmaceutically acceptable salt” is any non-toxic salt of a compound of the invention that, when administered to a recipient, can directly or indirectly provide a compound of the invention or a pharmaceutically active metabolite or residue thereof. Or an ester salt. As used herein, the term “pharmaceutically active metabolite or residue thereof” means that the metabolite or residue thereof is also a pharmaceutically active compound according to the present invention.

Pharmaceutically acceptable salts are well known in the art. For example, S.M. SM Berge et al. Describe pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 1977, 66, 1-19, incorporated herein by reference. Pharmaceutically acceptable salts of the compounds of this invention include those derived from suitable inorganic and organic acids and bases. Examples of pharmaceutically acceptable non-toxic acid addition salts are inorganic acids such as hydrochloric acid, bromic acid, phosphoric acid, sulfuric acid and perchloric acid, or organic acids such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid or malonic acid or in the art. Other methods used in, for example, salts of amino groups formed by using ion exchange. Other pharmaceutically acceptable salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropio Nate, digluconate, dodecyl sulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2- Hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, maleate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, Oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, piclay , Pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate salt and the like. Salts derived from suitable bases include alkali metal, alkaline earth metal, ammonium and N ⁺ (C _1-4 alkyl) ₄ salts. The invention also encompasses quaternization of any basic nitrogen-containing groups of the compounds described herein. Water soluble or fat soluble or dispersible products can be obtained by such quaternization. Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium and the like. Further pharmaceutically acceptable salts are, where appropriate, non-toxic ammonium, quaternary ammonium and counterions such as halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, lower alkyl sulfonates and aryls. Amine cations formed using sulfonates.

The compositions of the present invention may further comprise a pharmaceutically acceptable carrier, adjuvant or vehicle, and as used herein, all solvents, diluents or other liquid vehicles, dispersions or suspension preparations, to suit the particular dosage form desired, Surface active agents, isotonic agents, thickeners or emulsifiers, preservatives, solid binders, lubricants and the like. See Remington's Pharmaceutical Sciences, Sixteenth Edition, E.W. Martin (Mack Publishing Co., Easton, Pa., 1980) describes various carriers used to formulate pharmaceutically acceptable compositions and known techniques for their preparation. Except where conventional carrier media are incompatible with the compounds of the present invention, such as producing undesirable biological effects or otherwise interacting poorly with any other component (s) of the pharmaceutically acceptable composition. Its use is considered to be within the scope of the present invention. Some examples of materials that can be used as pharmaceutically acceptable carriers include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins, such as human serum albumin, buffer materials, eg For example, phosphates, glycine, sorbic acid or potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes, for example protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloids Acidic silica, magnesium trisilicate, polyvinyl pyrrolidone, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, wool, sugars such as lactose, glucose and sucrose; Starch such as corn starch and potato starch; Cellulose and its derivatives such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; Powdered tragacanth; malt; gelatin; Talc; Excipients such as cocoa butter and suppository waxes; Oils such as peanut oil, cottonseed oil; Sunflower oil; Rapeseed oil; olive oil; Corn oil and soybean oil; Glycols; Propylene glycol or polyethylene glycol; Esters such as ethyl oleate and ethyl laurate; Agar; Buffers such as magnesium hydroxide and aluminum hydroxide; Alginic acid; Pyrogen-free water; Isotonic saline; Ringer's solution; Ethyl alcohol and phosphate buffer solutions and other nontoxic compatible lubricants, such as sodium lauryl sulfate and magnesium stearate and colorants, mold release agents, coatings, sweeteners, spices and fragrances, preservatives and antioxidants It may be present in the composition as determined by.

The compositions provided by the present invention can be used in combination therapy, meaning that the compositions of the present invention can be administered simultaneously, before, or subsequently with one or more other desired therapeutic agents or medical procedures. The particular combination therapy (therapeutic agent or procedure) used in the combination therapy may take into account the compatibility of the desired therapeutic agent and / or procedure and the desired therapeutic effect to be achieved. The therapy used may achieve the desired effect on the same disorder (eg, a compound described herein may be administered concurrently with another therapeutic agent used to treat the same disorder), or a different effect (eg, It can be understood that the control of side effects) can be achieved.

For example, known agents useful for treating neurodegenerative disorders can be combined with the compositions of the present invention to treat neurodegenerative disorders such as Alzheimer's disease. Examples of these known agents useful to treat neurodegenerative disorders are, thus, but are not limited to, therapeutic agents for Alzheimer's disease, for example, donepezil, memantine (and related compounds as NMDA inhibitors), Exelon (Exelon ^® Acetylcholinesterase inhibitors, including; Therapeutics for Parkinson's disease, such as L-DOPA / carbidopa, entacapone, ropin roll, pramipexole, bromocriptine, pergolide, trihexefendil and amantadine; Agents for the treatment of multiple sclerosis (MS), such as beta interferon (eg, Avonex ^® and Rebif ^® ), Cofaxone ^® and Mitoxantrone; Rilusol and anti-Parkinson's agents. For a more comprehensive discussion of the latest therapies useful for treating neurodegenerative disorders, see the list of FDA-approved drugs on the site [http://www.fda.gov] and the Merck Manual, Seventeenth Ed. 1999, the entirety of which is incorporated herein by reference.

In another embodiment, the compounds of the present invention are combined with other agents useful for treating neurodegenerative disorders, such as Alzheimer's disease, wherein these agents are beta-secretase inhibitors, gamma-secretase inhibitors, aggregation Inhibitors, metal chelators, antioxidants and neuroprotective agents.

As used herein, the terms “compound”, “comprise” and related terms refer to simultaneous or subsequent administration of a therapeutic agent according to the invention. For example, the compounds of the present invention can be administered simultaneously or successively with another therapeutic agent in separate unit dose form or in single unit dose form. Accordingly, the present invention provides single unit dosage forms comprising a compound of formula (I), additional therapeutic agents and pharmaceutically acceptable carriers, adjuvants or vehicles.

Other examples of inhibitors of the present invention, for a therapeutic agent for asthma, without limiting, e.g., albuterol and SINGULAIR (Singulair ^®); Agents for the treatment of schizophrenia, such as Zyprexa, risperdal, Cerroquel and haloperidol; Anti-inflammatory agents such as corticosteroids, TNF blockers, IL-1 RA, azathioprine, cyclophosphamide and sulfasalazine; Immunomodulators and immunosuppressants such as cyclosporin, tacrolimus, rapamycin, mycophenolate mofetil, interferon, corticosteroids, cyclophosphamide, azathioprine and sulfasalazine; Nerve growth factors such as acetylcholinesterase inhibitors, MAO inhibitors, interferons, anticonvulsants, ion channel blockers, cardiovascular disease agents, such as beta-blockers, ACE inhibitors, diuretics, nitrates, calcium channel blockers And statins; Therapeutic agents for liver diseases such as corticosteroids, cholestyramine, interferon and antiviral agents; Blood disorder treatments such as corticosteroids, anti-leukemic agents and growth factors; And therapeutic agents for immunodeficiency disorders such as gamma globulin.

The amount of additional therapeutic agent present in the composition of the present invention will be less than the amount generally available for administration in a composition comprising the therapeutic agent only as the active agent. In certain embodiments, the amount of additional therapeutic agent in the composition of the present invention will range from about 50% to 100% of the amount generally present in a composition comprising the agent only as a therapeutically active agent.

In another embodiment, the methods of the present invention using a composition that does not include additional therapeutic agents include the additional step of separately administering additional therapeutic agents to the patient. When these additional therapeutic agents are administered separately, they may be administered to the patient, continuously or subsequently, prior to administration of the compositions of the present invention.

The pharmaceutically acceptable compositions of this invention may be administered orally, rectally, parenterally, subarachnoid, intravaginally, intraperitoneally, topically (by powder, ointment or mucus), depending on the severity of the disorder to be treated. It can be administered to the human and other animals of interest by spraying, such as in the cheeks. In certain embodiments, the compounds of the present invention contain from about 0.01 mg / kg to about 50 mg / kg of body weight, preferably from about 1 mg / kg to about 25 mg / kg of body weight, at least once a day, to achieve the desired therapeutic effect. It can be administered orally or parenterally at a dose level of.

Liquid dosage forms for oral administration include, but are not limited to, pharmaceutically acceptable emulsions, microemulsions, solvents, suspensions, syrups and elixirs. In addition to the active compounds, liquid dosage forms are inert diluents commonly used in the art, such as water or other solvents, solvating agents and emulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl Alcohols, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oils (especially cottonseed oil, peanut oil, corn oil, embryo oil, olive oil, castor oil and rapeseed oil), glycerol, tetrahydrofurfuryl Fatty acid esters of alcohols, polyethylene glycols and sorbitan, and mixtures thereof. In addition to inert diluents, oral compositions may also include auxiliaries such as wetting agents, emulsifiers and suspending agents, sweeteners, spices and fragrances.

Injectable preparations, for example, sterile injectable aqueous or oleaginous suspensions may be formulated according to the known art using suitable dispersing or wetting agents and suspending agents. Sterile injectable preparations may be, for example, sterile injectable solutions, suspensions or emulsions in a nontoxic parenterally acceptable diluent or solvent as a solution in 1,3-butanediol. Among the acceptable vehicles and solvents available are water, Ringer's solution (U.S.P.), and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose any bland fixed oil may be employed including synthetic monoglycerides or diglycerides. In addition, fatty acids such as oleic acid are used in the preparation of injectables.

Injectable formulations can be sterilized, for example, by incorporation into the sterilizing agent in the form of a sterile solid composition that can be dissolved or dispersed in sterile water or other sterile injectable media prior to use, by filtration through a bacteria-retaining filter. have.

In order to prolong the effects of the compounds of the invention, it is often desirable to slow the absorption of the compounds from subcutaneous or intramuscular injection. This can be achieved by the use of a liquid suspension of low water solubility crystalline or amorphous material. The rate of absorption of the compound then depends upon its rate of degradation, i.e., depending on crystal size and crystalline form. Alternatively, delayed absorption of the parenterally administered compound form is accomplished by dissolving or suspending the compound in an oil vehicle. Injectable depot forms are made by forming microencapsule matrices of the compound in biodegradable polymers such as polylactide-polyglycolide. Depending on the ratio of compound to polymer and the nature of the particular polymer employed, the rate of compound release can be controlled. Examples of other biodegradable polymers include poly (orthoesters) and poly (anhydrides). Depot injectable formulations are prepared by suturing the compound in liposomes or microemulsions that are compatible with body tissue.

Compositions for rectal or vaginal administration are preferably suppositories, which can be prepared by mixing the compounds of the present invention with a suitable non-irritating adjuvant or carrier such as cocoa butter, polyethylene glycol, or solid at ambient temperature but liquid at body temperature. It is a suppository wax that melts in the rectum or vagina to release the active compound.

Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In this solid dosage form, the active compound is one or more inert, pharmaceutically acceptable adjuvants or carriers such as sodium citrate or dibasic calcium phosphate and / or a) fillers or extenders such as starch, lactose , Sucrose, glucose, mannitol and silicic acid, b) binders such as carboxymethylcellulose, alginate, gelatin, polyvinylpyrrolidinone, sucrose and acacia, c) humectant, such as glycerol , d) disintegrants, for example agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates and sodium carbonate, e) solution retardants such as paraffin, f) absorption accelerators, for example Quaternary ammonium compounds, g) wetting agents such as cetyl alcohol and glycerol monostearate, h) absorbents such as kaolin and bentonite clay and i) suspending agents such as talc, Mixed with calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate and mixtures thereof. In the case of capsules, tablets and pills, the dosage form may comprise a buffer.

Solid compositions of a similar form may also be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or lactose and high molecular weight polyethylene glycols and the like. Solid dosage forms of tablets, dragees, capsules, pills and granules can be prepared with coatings and shells such as enteric coatings and other coatings well known in the pharmaceutical formulation art. They may optionally comprise an opacifying agent and may also be compositions which release only the active ingredient (s) or preferably in certain parts of the small intestine in an optionally delayed manner. Examples of embedded compositions that can be used include polymeric substances and waxes. Solid compositions of a similar form may also be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or lactose and high molecular weight polyethylene glycols and the like.

The active compound may be in microencapsulated form with one or more excipients described above. Solid dosage forms of tablets, dragees, capsules, pills and granules can be prepared with coatings and shells such as enteric coatings, release controlling coatings and other coatings well known in the pharmaceutical formulation art. In this solid dosage form, the active compound may be premixed with one or more inert diluents, such as sucrose, lactose or starch. Such dosage forms may in general practice comprise further substances other than inert diluents such as tableting suspending agents and other tableting aids such as magnesium stearate and microcrystalline cellulose. In the case of capsules, tablets and pills, the dosage form may comprise a buffer. They may optionally comprise an opacifying agent and may also be compositions which release only the active ingredient (s) or preferably in certain parts of the small intestine in an optionally delayed manner. Examples of embedded compositions that can be used include polymeric substances and waxes.

Dosage forms for topical or transdermal administration of a compound of this invention include ointments, pastes, creams, lotions, gels, powders, solvents, sprays, inhalants or patches. The active ingredient is mixed under sterile conditions with a pharmaceutically acceptable carrier and the necessary preservative or buffer that may be necessary. Ophthalmic formulations, ear drops and eye drops are also contemplated as being within the scope of this invention. In addition, the present invention contemplates the use of transdermal patches having the additional advantage of providing controlled delivery of the compound to the body. Such dosage forms can be made by dissolving or dispensing the compound in the proper medium. Absorption enhancers can also be used to increase the influx of compounds through the skin. The rate can be controlled by providing a rate controlling membrane or by dispersing the compound in a polymer matrix or gel.

In some embodiments, the present invention provides a composition comprising a compound of Formula I, II, III, IVa, IVb, IVc, V, VI, VII, VIII, IX, or X in an amount of about 1% to about 99% by weight. to provide. In another embodiment, a composition comprising a compound of Formula (I), (II), (III), (IVa), (IVb), (IVc), (V), (VI), (VII), (VIII), (IX), or (X) is a different component of the black cohosh root over the total area of the HPLC chromatogram. Up to about 10.0 area% HPLC. In another embodiment, a composition comprising a compound of Formula (I), (II), (III), (IVa), (IVb), (IVc), (V), (VI), (VII), (VIII), (IX), or (X) is a different component of the black cohosh root over the total area of the HPLC chromatogram. Up to about 8.0 area% HPLC, and in another embodiment, up to about 3 area%.

Use of Compounds and Pharmaceutically Acceptable Compositions

Compounds of the invention are useful for modulating and / or inhibiting amyloid-beta (1-42) peptide production in a patient. Accordingly, the compounds of the present invention are useful for treating or lessening the severity of disorders associated with amyloid-beta (1-42) peptide production in patients.

The compounds, extracts, and compositions according to the methods of the invention may be administered using an amount and route of administration effective for treating or lessening the severity of a neurodegenerative disorder. The exact amount required may vary from subject to subject, depending on the subject's gender, age and general condition, severity of infection, particular agent, method of administration thereof and the like.

In certain embodiments, the invention provides a method of administering to a patient a compound of Formula I, II, III, IVa, IVb, IVc, V, VI, VII, VIII, IX, or X or a pharmaceutically acceptable composition comprising the compound. A method of modulating and / or inhibiting amyloid-beta (1-42) peptide production in a patient is provided. In another embodiment, the present invention comprises administering to a patient a compound of Formula (I), (II), (III), (IVa), (IVb), (IVc), (V), (VI), (VII), (VIII), (IX) or (X), or a pharmaceutically acceptable composition thereof. Methods of selectively regulating and / or inhibiting amyloid-beta (1-42) peptide production in a patient are provided. In another embodiment, the invention comprises administering to a patient a compound of Formula (I), (II), (III), (IVa), (IVb), (IVc), (V), (VI), (VII), (VIII), (IX) or (X), or a pharmaceutically acceptable composition thereof. , A method of reducing amyloid-beta (1-42) peptide levels in a patient. In another embodiment, the present invention comprises contacting a cell with a compound of Formula (I), (II), (III), (IVa), (IVb), (IVc), (V), (VI), (VII), (VIII), (IX) or (X). 42) A method of reducing peptide levels is provided. Another embodiment comprises amyloid-beta (1-40) peptide in a cell, comprising contacting the cell with a compound of Formula I, II, III, IVa, IVb, IVc, V, VI, VII, VIII, IX or X Provided are methods for reducing amyloid-beta (1-42) in cells without substantially reducing levels. Another embodiment includes contacting a cell with a compound of Formula (I), (II), (III), (IVa), (IVb), (IVc), (V), (VI), (VII), (VIII), (IX), or (X), to inhibit amyloid-beta (1-42) in a cell. A method of removing and increasing one or more amyloid-beta (1-37) and amyloid-beta (1-39) in a cell is provided.

As used herein, the term "reducing" or "reducing" refers to the amyloid-beta in the absence of administration of a compound of Formula (I), (II), (III), (IVa), (IVb), (IVc), (V), (VI), (VII), (VIII), (IX) or (X). It means that the amount of amyloid-beta achieved by administration of a compound of formula (I), (II), (III), (IVa), (IVb), (IVc), (V), (V), (VII), (VIII), (IX) or (X) is relatively reduced. In one example, the decrease in amyloid-beta (1-42) is amyloid-beta (1-42) in the presence of a compound of formula (I), (II), (III), (IVa), (IVb), (IVc), (V), (VI), (VII), (VIII), (IX) or (X). ) Is less than the amount of amyloid-beta (1-42) in the absence of compounds of formula (I), (II), (III), (IVa), (IVb), (IVc), (V), (VI), (VII), (VIII), (IX) or (X).

In another embodiment, the invention comprises administering to a patient a compound of Formula (I), (II), (III), (IVa), (IVb), (IVc), (V), (VI), (VII), (VIII), (IX) or (X), or a pharmaceutically acceptable composition thereof. A method of selectively reducing amyloid-beta (1-42) peptide levels in a patient is provided. In certain embodiments, the invention comprises administering to a patient a compound of Formula I, II, III, IVa, IVb, IVc, V, VI, VII, VIII, IX or X, or a pharmaceutically acceptable composition thereof, Provided are methods for reducing amyloid-beta (1-42) peptide levels in a patient without substantially reducing amyloid-beta (1-40) peptide levels.

In certain embodiments, the invention comprises administering to a patient a compound of Formula I, II, III, IVa, IVb, IVc, V, VI, VII, VIII, IX or X, or a pharmaceutically acceptable composition thereof, Methods of reducing amyloid-beta (1-42) peptide levels and increasing one or more amyloid-beta (1-37) and amyloid-beta (1-39) in a patient are provided.

As used herein, the term "increasing" or "increasing" in relation to the amount of amyloid-beta means the administration of a compound of formula I, II, III, IVa, IVb, IVc, V, VI, VII, VIII, IX or X (Or contacting a cell with a compound of Formulas I, II, III, IVa, IVb, IVc, V, VI, VII, VIII, IX, or X) in the absence of the amount of Formula I, II, III Administration of a compound of formula (IVa, IVb, IVc, V, VI, VII, VIII, IX or X (or a cell with formula (I), II, III, IVa, IVb, IVc, V, VI, VII, VIII, IX or X) Means a relatively increased amount of amyloid-beta achieved by contact with a compound of). In one example, the increase in amyloid-beta (1-37) is amyloid-beta (1-37) in the presence of a compound of formula (I), (II), (III), (IVa), (IVb), (IVc), (V), (VI), (VII), (VIII), (IX) or (X). ) Is greater than the amount of amyloid-beta (1-37) in the absence of compounds of formulas I, II, III, IVa, IVb, IVc, V, VI, VII, VIII, IX or X. For example, the relative amounts of amyloid-beta (1-37) and amyloid-beta (1-39) may be increased by increasing the production of amyloid-beta (1-37) and amyloid-beta (1-39) or longer amyloids. It can be increased by reducing the production of beta peptides such as amyloid-beta (1-40) and / or amyloid-beta (1-42). In addition, the terms "increasing" or "increasing" as used herein with respect to the amount of amyloid-beta are compounds of formula I, II, III, IVa, IVb, IVc, V, VI, VII, VIII, IX or X It is understood to mean an absolute increase in the amount of amyloid-beta achieved by administration of. Thus, in certain embodiments, the invention comprises administering to a patient a compound of Formula I, II, III, IVa, IVb, IVc, V, VI, VII, VIII, IX, or X or a pharmaceutically acceptable composition thereof. Thus, a method of increasing the absolute level of one or more of amyloid-beta (1-37) and amyloid-beta (1-39) is provided. In another embodiment, the invention comprises administering to a patient a compound of Formula I, II, III, IVa, IVb, IVc, V, VI, VII, VIII, IX or X or a pharmaceutically acceptable composition thereof, Provided are methods for increasing the level of either amyloid-beta (1-37) or amyloid-beta (1-39), wherein the increase is a longer amyloid-beta peptide, for example amyloid-beta (1). -40) and / or amyloid-beta (1-42), or relative to the amount of total amyloid-beta.

Those skilled in the art will understand that the overall proportion of amyloid-beta peptides is significant when the selective reduction of amyloid-beta (1-42) is particularly advantageous. In certain embodiments, the compounds of the present invention reduce the overall ratio of amyloid-beta (1-42) peptides to amyloid-beta (1-40) peptides. Accordingly, another aspect of the present invention includes administering to a patient a compound of Formula (I), (II), (III), (IVa), (IVb), (IVc), (V), (VI), (VII), (VIII), (IX), or a pharmaceutically acceptable composition thereof. The present invention provides a method of reducing the ratio of amyloid-beta (1-42) peptide to amyloid-beta (1-40) peptide in a patient. In certain embodiments, the ratio of amyloid-beta (1-42) peptide to amyloid-beta (1-40) peptide decreases from the range of about 0.1 to about 0.4 to the range of about 0.05 to about 0.08.

In another embodiment, the present invention comprises contacting a cell with a compound of Formula (I), (II), (III), (IVa), (IVb), (IVc), (V), (VI), (VII), (VIII), (IX), or (X) to form amyloid-beta (1- 40) A method of reducing the ratio of amyloid-beta (1-42) peptide to peptide is provided. In certain embodiments, the ratio of amyloid-beta (1-42) peptide to amyloid-beta (1-40) peptide decreases from the range of about 0.1 to about 0.4 to the range of about 0.05 to about 0.08.

According to one aspect, the invention comprises administering to a patient a compound of Formula I, II, III, IVa, IVb, IVc, V, VI, VII, VIII, IX or X or a pharmaceutically acceptable composition thereof Provided are methods for treating or lessening the severity of disorders associated with amyloid-beta (1-42) peptides. Such disorders include neurodegenerative disorders such as Alzheimer's disease, Parkinson's disease and Down's syndrome.

Such disorders also include inclusion body myositis (sedimentation of A-beta in peripheral muscle, causing peripheral neuropathy), cerebral amyloid angiopathy (amyloid in blood vessels in the brain), and mild cognitive impairment.

“High A-Beta42” is a measurable condition that occurs earlier than symptomatic disease, particularly in familial patients, based on plasma, CSF measurements, and / or genetic screening. This concept is similar to the relationship between elevated cholesterol and heart disease. Accordingly, another aspect of the present invention comprises administering to a patient a compound of Formula I, II, III, IVa, IVb, IVc, V, VI, VII, VIII, IX or X, or a pharmaceutically acceptable composition thereof. Thus, a method of preventing disorders associated with elevated amyloid-beta (1-42) peptides is provided.

In another embodiment, the invention comprises administering to a patient a compound of Formula I, II, III, IVa, IVb, IVc, V, VI, VII, VIII, IX or X or a pharmaceutically acceptable composition thereof, Provided are methods for treating diseases in which A-beta amyloidosis may be a underlying aspect or a factor present or exacerbated simultaneously.

In another aspect, the invention comprises administering to a patient a compound of Formula I, II, III, IVa, IVb, IVc, V, VI, VII, VIII, IX, or X or a pharmaceutically acceptable composition thereof. Lewy body dementia in patients (associated with precipitation of alpha-synuclein from cognitive neurons to Lewy body, and a-synuclein more commonly associated with deposition in motor neurons and pathogenesis of Parkinson's disease) , Cataracts (a-beta are condensed into the ocular lens), tauopathy (e.g., temporal lobe dementia), Huntington's disease, ALS / Lou Gehrig's disease, type 2 diabetes (IAPP aggregation in islet islands is dependent on the size and sequence of A-beta Similar and type 2 diabetes increases the risk of dementia), transthyretin amyloid disease (TTR, one example of this disease is heart muscle that contributes to cardiomyopathy), prion diseases (Creutzfeldt-Jakob disease, Gerstmann-Schute) Reusler-Shinker Syndrome, Including fatal familial insomnia and kuru] and CJD.

In another embodiment, the invention comprises administering to a patient a compound of Formula I, II, III, IVa, IVb, IVc, V, VI, VII, VIII, IX or X or a pharmaceutically acceptable composition thereof, Provided are methods for treating or lessening the severity of Alzheimer's disease in a patient.

Without wishing to be bound by any particular theory, the compounds of the present invention are considered to be modulators of gamma-secretase that selectively reduce the level of amyloid-beta (1-42). Accordingly, another embodiment of the present invention comprises administering to a patient a compound of Formula I, II, III, IVa, IVb, IVc, V, VI, VII, VIII, IX or X or a pharmaceutically acceptable composition thereof. Thus, a method of controlling gamma-secretase in a patient is provided. In certain embodiments, the compounds of the invention are inhibitors of gamma-secretase. The method is useful for treating or lessening the severity of disorders associated with gamma-secretase. Such disorders include but are not limited to neurodegenerative disorders such as Alzheimer's disease.

Notch / delta signaling pathways are highly conserved across species and widely used during the development of both vertebrates and invertebrates to regulate cell death in developing embryos. Gaiano and Fishell, "The Role of Notch in Promoting Glial and Neural Stem Cell Fates "Annu. Rev. Neurosci. 2002, 25: 471-90. Notches interact with gamma-secretase complexes and with other protein variants and signaling pathways. Notch 1 competes with amyloid precursor protein for gamma-setletase, and the activity of the Notch signaling pathway down-regulates PS-1 gene expression. Lleo et al, "Notch1 Competes with the Amyloid Precursor Protein for g -Secretase and Down-regulates Presenilin-1 Gene Expression "Journal of Biological Chemistry 2003, 48: 47370-47375. Notch receptors synergize with T cell receptors and develop by gamma-secretase action, thus maintaining peripheral T cell activity. Notch 1 can directly regulate Tbx21 via a complex formed on a Tbx21 promoter (Minter et al., "Inhibitors of g-secretase block in vivo and in vitro T helper type 1 polarization by preventing Notch upregulation). of Tbx21, "Nature Immunology 2005, 7: 680-688." In vitro, gamma-secretase inhibitors abolished expression of Notch, interferon-gamma and Tbx21 in TH1-polarized CD4 + cells. In vivo, gamma-secretase inhibitors substantially delayed the development of TH1-mediated disease in a mouse experimental autoimmune encephalomyelitis model of multiple sclerosis, which indicated the availability of such compounds for the treatment of TH1-mediated autoimmunity. Suggested [Id]. Inhibition of gamma-secretase can alter lymphocyte production and intestinal cell differentiation [Wong et al., "Chronic Treatment with the g-Secretase Inhibitor LY-411,575 Inhibits b-Amyloid Peptide Production and Alters Lymphopoiesis and Intestinal Cell Differentiation "Journal of Biological Chemistry 2004, 26: 12876-12882], which includes the induction of goblet cell metaplasia [Milano et al.," Modulation of Notch Processing by g-Secretase Inhibitors Causes Intestinal Goblet Cell Metaplasia and Induction of Genes Known to Specify Gut Secretory Lineage Differentiation "Toxicological Sciences 2004, 82: 341-358.

Strategies that can alter amyloid precursor protein ("APP") and reduce the production of pathogenic forms of amyloid-beta without affecting the notch process are highly desirable. In addition, as described above, inhibition of gamma-secretase has been found to be useful for inhibiting Th cells in vitro and in vivo and thus for treating disorders associated with Th1 cells. Th1 cells, in some instances, are associated with the pathogenesis of various organ-specific autoimmune disorders, Crohn's disease, Helicobacter pylori-induced peptic ulcer, acute renal allograft rejection and unexplained habitual miscarriage.

In one embodiment, the invention relates to a method of inhibiting the formation of Th1 cells in a patient, comprising administering to the patient a compound of the invention or a composition comprising said compound. In certain embodiments, the invention provides a compound of Formula I, II, III, IVa, IVb, IVc, V, VI, VII, VIII, IX or X, or a pharmaceutical composition comprising said compound prepared according to the method of the invention. One or more including irritable bowel disorders, Crohn's disease, rheumatoid arthritis, psoriasis, Helicobacter pylori-induced peptic ulcer, acute renal allograft rejection, multiple sclerosis or systemic lupus erythematosus, including administering an acceptable composition to a patient Provided are methods for treating autoimmune disorders.

In certain embodiments, the invention administers to a patient a compound of Formula I, II, III, IVa, IVb, IVc, V, VI, VII, VIII, IX, or X or a pharmaceutically acceptable composition comprising said compound Provided are methods for modulating and / or inhibiting amyloid-beta peptide production in a patient without affecting the Notch process, including.

In certain embodiments, the invention administers to a patient a compound of Formula I, II, III, IVa, IVb, IVc, V, VI, VII, VIII, IX, or X or a pharmaceutically acceptable composition comprising said compound Including a method of inhibiting amyloid-beta (1-42) peptide production in a patient, without affecting the notch process.

In certain embodiments, the invention administers to a patient a compound of Formula I, II, III, IVa, IVb, IVc, V, VI, VII, VIII, IX, or X or a pharmaceutically acceptable composition comprising said compound Reducing amyloid-beta (1-42) peptide levels and increasing one or more of amyloid-beta (1-37) and amyloid-beta (1-39) in patients without affecting the notch process, including Provide a method.

Accordingly, another aspect of the present invention comprises administering to a patient a compound of Formula I, II, III, IVa, IVb, IVc, V, VI, VII, VIII, IX or X, or a pharmaceutically acceptable composition thereof. Thus, a method is provided for reducing the ratio of amyloid-beta (1-42) peptide to amyloid-beta (1-40) peptide in a patient without affecting the notch process. In certain embodiments, the ratio of amyloid-beta (1-42) peptide to amyloid-beta (1-40) peptide is reduced from the range of about 0.1 to about 0.4 to the range of about 0.05 to about 0.08.

The compounds of the present invention are preferably formulated in dosage unit form for ease of administration and uniformity of dosage. As used herein, the expression “dose unit form” means a physically discrete unit of preparation suitable for the patient to be treated. However, it is understood that the total daily use of the compounds and compositions of the present invention can be determined by the attending physician within the scope of sound medical judgment. Specific effective amount levels for a particular patient or organism may include the disorder to be treated and the severity of the disorder; The activity of the specific compound employed; The specific composition employed; The age, body weight, general health, sex and diet of the patient; The time of administration, route of administration, and rate of excretion of the specific compound employed; Duration of treatment; It may be dependent on various factors including factors well known in the medical art, such as drugs used in combination or co-administration with the particular compound employed. As used herein, the term "patient" means an animal, preferably a mammal, most preferably a human.

The black cohosh extract used in the separation protocol described below was purchased by custom order from Boehringer Ingelheim Nutriceutical. The extract is substantially equivalent to the USP formulation of the black cohosh extract, wherein about 50% aqueous ethanol is used to extract the powdered roots, and then concentrated to dryness.

As used herein, the compound numbers described below correspond to the following compounds.

Compound 1: β-D-xylopyranoside, (3,12,16,23R, 24R, 25S, 26S) -12- (acetyloxy) -16,23: 23,26: 24,25-triepoxy -26-hydroxy-9,19-cyclolanostan-3-yl.

Also known as "actane." C37H56O11; Molar weight: 676.83; Registration No. 18432-44-9.

           One

Compound 2: simigenol 3-β-D-xylopyranoside; C 35 H 56 O 9, molar weight: 620.81; Registration No. 27994-11-2.

           2

Compound 3: simigenol 3-α-L-arabinoside. C 35 H 56 O 9, molar weight: 620.81; Registration No. 256925-92-5.

           3

Compound 4: 24-O-acetylhydroshengmanol 3-β-D-xylopyranoside. C 37 H 60 O 11, molar weight 680.87; Registration No. 78213-32-8.

            4

Compound 5: 24-O-acetylhydrogenhengnol 3-α-L-arabinofyranoside. C 37 H 60 O 11, molar weight: 680.87.

             5

Compound 6: 24-O-acetylhydrogenhenolmanol 3-β-D-xylopyranoside (delta-16,17) -enol ether. C 37 H 58 O 10, molar weight: 662.85.

             6

Compound 7: 24-O-acetylhydrogenhengnol 3-α-L-arabinofyranoside (delta-16,17) -enol ether. C 37 H 58 O 10, molar weight: 662.85.

           7

Compound 8: 24-Epi-24-O-acetylhydrogenhengmanol 3-β-D-xylopyranoside. C 37 H 60 O 11, molar weight: 680.87.

            8

Compound 9: 24-Epi-24-O-acetylhydrogenhengnol 3-β-D-xylopyranoside (delta-16,17) -enol ether. C 37 H 58 O 10, molar weight: 662.85.

            9

Separation Protocol 1

Flash Column Chromatography

Black cohosh extract (15.6 g) was suspended in 150 mL of 4-to-1 (v / v) methanol-water mixture at 25 ° C. Using a mechanical stirrer, the resulting slurry was vigorously stirred at this temperature for 30 minutes, resulting in a brown emulsion. To the emulsion was added 51 g of silica gel (ICN silica 32-63 60 Pa) with continued stirring. The mixture was concentrated using a rotary evaporator in vacuo at 25 ° C. until a very homogeneous beige to brown powder was obtained. The material was column chromatographed on silica gel (ICN silica 32-63 60 Hz) using a glass column with an internal diameter of 50 mm and a length of 60 cm.

In the formulation for column chromatography, silica gel was poured into 500 mL of 20-to-1 dichloromethane-methanol mixture and the resulting slurry was poured into a glass column. The silica gel was allowed to settle for 30 minutes and covered with a layer of sand 1 cm thick. Subsequently, the extract absorbed by silica was poured into a 20-to-1 dichloromethane-methanol mixture and the resulting slurry was poured into a sand layer on top of the column. The silica column was then eluted with the following solvent mixture under 0.4 bar (argon) pressure:

1.0 ml dichloromethane-methanol 20-to-1 then

770 mL dichloromethane-methanol 10-to-1 then

800 mL dichloromethane-methanol 7-to-1, then

550 mL dichloromethane-methanol 5-vs-1.

200 ml 8 fractions (labeled sat14-0 to sat14-7) were collected followed by 100 ml 11 fractions (labeled sat14-8 to sat14-18). All fractions were analyzed by thin layer chromatography (TLC) using a Bakerflex silica plate and eluted with a 5-to-1 dichloromethane-methanol solvent mixture. After development, the silica gel plates were colored with an anisealdehyde colorant. Based on the results of TLC analysis, fractions sat14-9 to sat14-12 were evaporated to dryness in vacuo at 25 ° C., these 10 mg samples were analyzed by ¹ H-NMR spectrometer using CD ₃ OD as solvent. See FIGS. 1 and 2, respectively. Since the signal is characteristic of compound 7 and compound 6, the spectra are analyzed in relation to the presence of wide multinomial at 2.53 ppm and 2.2 Hz doublet at 4.86 ppm. Further dqf-COSY spectra of these four samples confirm that the signals 2.53 ppm and 4.86 ppm actually correspond to compound 7 and compound 6. From the ¹ H-NMR spectrum of fraction sat14-10, it was concluded that although the sample contained the highest concentrations of compound 7 and compound 6, some small amounts of these compounds could be detected in fraction sat14-9. While fraction sat14-11 appears to contain trace amounts of compound 7 and compound 6, these compounds are not detected in fraction sat14-12. Based on these results, fraction sat14-10 was selected for further purification via HPLC. Alternatively, fraction sat14-9 can be used to obtain additional amounts of the required compound 4 to compound 7.

The main component of fraction sat14-10 is Actin (1) (JNP 2002, 65, 601-605), which crystallized from the methanol solution of this fraction. Pure actin is obtained through recrystallization. The main components of the fraction sat14-11 are simigenol beta-D-xylpyranoside (2) and simigenol alpha-L-arabinoside (3), which are about 2: 1 (see JNP 2000, 65, 905). -910 and 1391-1397) and recrystallized from this fraction as a mixture.

Reversed Phase HPLC Fractionation on C-18 Columns

Fraction sat14-10 was dissolved in 3.5 ml methanol. The solution was subjected to an AGILENT comprising a SUPELCO Discovery RP-18 column (25 cm long, 10 mm inner diameter), and an auto-injector and a diode array detector used for wavelength detection between 190 and 400 nm. ) Fractionated by HPLC using a 1100 series HPLC system. The solvent concentration gradient was used as a straight line reduction in water content from 20 minutes to 100% methanol, starting with 30% (v / v) water in methanol for the first 2 minutes. After 2 minutes in 100% methanol, the water content was increased to 30% and maintained at this concentration for an additional 8 minutes. For isolation of the entire sample sat14-10, 100 injections of 35 μl each are required. Nine fractions were collected and labeled with sat15-1 to sat15-9. See FIGS. 3 and 4, respectively. Compounds 4-7 were eluted in fractions sat15-1, 15-2, 15-4 and 15-5. ¹ H NMR spectra of the fractions sat15-1, 15-2, 15-4 and 15-5 are shown in FIGS. 4A and 4B.

Reverse Phase HPLC Fractionation on C-8 Columns for Separation of Compounds 6, 4 and 9

Fraction sat15-5 was dissolved in 1.5 ml methanol. The solution was fractionated by HPLC using a SUPELCO supelcosil LC-8 column (25 cm long, 10 mm inner diameter), and the Agilent 1100 Series HPLC system described above. The solvent concentration gradient was used as a linear reduction in water content from 20 minutes to 100% methanol, starting with 40% (v / v) water in methanol for the first 2 minutes. After 2 minutes in 100% methanol, the water content was increased to 40% and maintained at this concentration for an additional 8 minutes. For isolation of the entire sample sat15-5, 30 μl 50 injections each are required. Five fractions were collected and labeled with sat16-1 to sat16-5 (FIG. 5). Compound 6 eluted in fraction sat16-3, while compound 4 eluted in fraction sat16-1. A small amount of pure compound 9 was obtained in fraction sat15-5. FIG. 6 shows a ¹ H NMR spectrum of 9.8 mg of Compound 6 in 98% purity obtained.

Reversed Phase HPLC Fractionation on C-8 Columns for Separation of Compound 8

Fraction sat15-8 was dissolved in 0.65 mL methanol. The solution was fractionated by HPLC using a Sufelco Sufelcosyl LC-8 column (25 cm long, 10 mm inner diameter), and the Agilent 1100 Series HPLC system described above. The solvent concentration gradient was used as a linear reduction in water content from 20 minutes to 100% methanol, starting with 40% (v / v) water in methanol for the first 2 minutes. After 2 minutes in 100% methanol, the water content was increased to 40% and maintained at this concentration for an additional 8 minutes. Seven fractions were collected and labeled with sat18-1 to sat18-7. Compound 8 was eluted at fraction sat18-6. NMR-spectrometry analysis, including the NOESY spectrum, shows that compound 8 is interconverted with the corresponding ketone in methanol solution. Dilute methanol solution comprises about 4% ketone and 96% hemiacetal form.

Reversed Phase HPLC Fractionation on C-8 Columns for Separation of Compounds 7 and 5

Fraction sat15-2 was dissolved in 0.5 ml methanol. The solution was fractionated by HPLC using a Sufelco Sufelcosyl LC-8 column (25 cm long, 10 mm inner diameter), and the Agilent 1100 Series HPLC system described above. The solvent concentration gradient was used as a linear reduction in water content from 20 minutes to 100% methanol, starting with 40% (v / v) water in methanol for the first 2 minutes. After 2 minutes in 100% methanol, the water content was increased to 40% and maintained at this concentration for an additional 8 minutes. Five fractions were collected and labeled as sat19-3 to sat19-7. Pure compound 7 was obtained in fraction sat19-7, while pure compound 5 was obtained in fraction sat19-5.

Separation Protocol 2

Alternative separation / purification protocols are described below to separate compound 6. Those skilled in the art will understand that while separating compound 6, other compounds of the invention are abundant and / or separated by this process. A summary of this separation process is shown in FIG.

The purification protocol uses the following device.

(a) Hitachi HPLC system with diode array detector (DAD)

(b) Nova Prep ™ 8000 semi-preparative HPLC with Remote PC Controller using LC ReSponder ™ Application Software

(c) Hitachi UV Detector L-7400

(d) Sedex 55 Evaporative Light Scattering (ELSD) Detector

(e) 75L Biotage Silica Column (KP-Sil; P / N FKO-1107-19073; Lot No. 027075L)

(f) 75L Biotage C18 column (backkerbond, 40μ)

(g) 75S Biotage C18 Column (Bydaq, 40μ)

(h) Analytical HPLC column: Phenomenex Luna C18, 3μ, 4.6 x 100 mm

(i) Semi-preparative HPLC column: Phenomenex Luna C8 HPLC column, 20 × 250 mm

(j) Semi-preparative HPLC column: YMC AQ C18 HPLC column, 21.2 × 250 mm and

(k) preparative HPLC column: preparative HPLC column for ES Industries C18; 5 x 25 cm.

The analytical method used to determine the purity of compound 6 is as follows.

Column: Phenomenex Luna C18, 3μ, 4.6 × 100 mm

Mobile phase: isocratic elution by A. 35% acetonitrile; B. 30% Nanoopure water with 0.05% acetic acid; And C. 35% MeOH

Flow rate: 1ml / min

Detection: 205, 230 nm, DAD; And ELSD

Running time: 8 minutes

Column temperature: 32 ℃

This method was used for the analysis of extracts, fractions and final products. Compound 6 eluted at about 5.5 minutes under the above conditions.

50 g of crude black cohosh extract ("BCE") was fractionated in a Biotage silica cartridge (7.5 x 30 cm). After filling, the cartridge was eluted with 5% MeOH / DCM (10 L) and 10% MeOH / DCM (5 L) and 500 mL fractions were collected. The flow rate is 150-200 ml / min. HPLC (UV at 230 nm) related compound 6 is present in fraction 23 (2.6 g) and fraction 24 (2.3 g). Fraction 23 (F23) was selected for further purification on a semi-preparative C8 column.

Ten runs were carried out to yield approximately 10 mg of compound 6. 50 mg of F23 in 0.3 mL MeOH was charged to a Phenomenex Luna C-8 (21.2 × 250 mm, 10 μ, 100 μs) semi-preparation column. The column was eluted at a flow rate of 9.9 mL / min with 70% MeOH in H ₂ O while monitoring with UV at 205 nm. Peaks eluting at 35 and 38 minutes were collected separately as shown in the semi-preparative HPLC traces (FIG. 8).

Fractions collected for peaks at 35 minutes from 10 runs were filled and the solvent was evaporated at ambient temperature. The solid obtained was dried in a lyophilizer to yield 10.3 mg of compound 6 (2609-165-7). While each fraction of HPLC shows only one major peak (FIG. 10), HPLC of product 2609-165-7 (FIG. 9) has a polar impurity peak (11.3%) with a retention time (RT) at 4.5 minutes. appear. Certainly compound 6 was converted to a more polar compound slowly during the process. The more polar compound shows proton NMR (Fig. 12) of impurities isolated at 4.5 min in which there is no singlet for SSI-MS and acetyl methyl showing a strong [M + Na] ⁺ peak at m / z 643 (Fig. 11). As evident from), it was found to be a deacetyl derivative of compound 6.

Some stability experiments with compound 6 show that deacetylation occurs in a slightly basic MeOH solution. However, it is stable in slightly acidic solutions. Thus, 2609-165-7 was retreated on a Luna C8 column using 70% MeOH / 30% water with 0.05% AcOH as eluent to yield 3.4 mg of compound 6 (2609-172-11). HPLC chromatograms of 2609-172-11 are shown in FIG. 13. Proton NMR (in CD ₃ OD) and SSI-MS are shown in FIGS. 14 and 15.

In another process, 250 g of black cohosh extract (BCE) was stirred with 1250 mL of MeOH for 1 hour at room temperature in a beaker. Although not all solids are dissolved, HPLC analysis of the filtrate shows that all compound 6 in the starting extract is dissolved (about 250 mg). Nevertheless, the unfiltered mixture was added to 750 g of silica gel (ICN, 60-200 μ) in a 5 L round bottom flask. MeOH was removed from the rotary evaporator with the aid of vacuum with a dry powder weighed 1,100 g with 9% residual MeOH.

The dried BCE in the silica formulation was divided into four portions of approximately 270 g each. The mixture is charged into SIM and first washed with 500-600 mL of methylene chloride to remove nonpolar material and residual MeOH. The SIM was connected to a 75L silica column (KP-Sil; P / N FKO-1107-19073; Lot No. 027075L; 7.5 × 25 cm or 1750 mL). The main column was radially compressed at 60 psi. The system was eluted with acetone at a flow rate of 100 mL / min and 500-1,000 mL fractions were collected. After eluting compound 6, the column was washed with 1.0 L of MeOH and rebalanced with 2 L of acetone. After approximately 900-1000 mL of acetone elutes from the column in Fraction 1 and Fraction 2, it is observed that Compound 6 elutes mainly in Fraction 3 (1,000 mL). Four runs first yielded approximately 224 mg of compound 6. A second batch of starting material for silica biotage was prepared from 100 g of BCE and 500 ml of MeOH and 300 g of silica. Two additional Biotage runs (5 and 6) were performed similar to the first four runs with this starting material which yielded an additional 93 mg of Compound 6. The product pools from six runs were combined and evaporated to dry solids under reduced pressure.

Solid (90 g) dried from silica biotage was dissolved in 720 mL of MeOH and 480 mL of H ₂ O was added slowly with stirring. Some dark tar-shaped solids were settled and removed from the filter. The cloudy filtrate was charged to a 75 L (7.5 × 25 cm) Backkerbond 60 mm 3, 40 μ Biotage C18 column. After filling (which is negatively tested from compound 6), the column is washed with 5 L of 60% (v / v) MeOH / H ₂ O followed by 4 L of 70% MeOH / H ₂ O, followed by 80% MeOH / Eluted with 4 L of H ₂ O. After eluting, the column was washed with 2 L of MeOH. The flow rate is about 60 ml / min in total, and the MeOH / H ₂ O mobile phase contains 0.05% acetic acid to prevent degradation of compound 6. The product pool (4 L) was concentrated under reduced pressure until essentially all the MeOH was removed and the precipitated solid obtained was collected in a Buchner funnel and dried at room temperature with the help of high vacuum.

Tar-type solids removed by filtration from the first large-scale C18 feed formulation and containing about 32 mg of compound 6 were dissolved in 2 L of MeOH wash from large-scale experiments, which included about 22 mg of compound 6. The mixture was evaporated to 1 L and mixed with 0.67 L water. Some tar-like solids were allowed to settle and collected in a filter, dissolved in 200 mL of MeOH and mixed with 134 mL of water. The mixture was also filtered to remove a small amount of tar and the filtrate was combined with the first filtrate and packed into a 75S (7.5 × 9.0 cm; 400 mL) Vydac 300 mm 3, 40 μ Biotage C18 column. The column was washed with 1 L of 60% MeOH / H ₂ O and ₂ L of 70% MeOH / H ₂ O and eluted with 1 L of 80% MeOH / H ₂ O (mobile phase also contained 0.05% acetic acid). The product pool was evaporated and the solids collected by filtration similar to large scale biotage experiments.

The first product pool (16.69 g) was mixed with 70 mL MeOH from a C18 biotage column. The mixture was sintered and the precipitate was removed by filtration. The filtrate was chromatographed with 70% MeOH / 30% water containing 0.05% AcOH as eluent at a flow rate of 177 mL / min on an Yees Industries Chromegabond WR C18 column (run 5 times, 14 mL each). . Fractions from 6-14 minutes of each run were combined and evaporated to remove MeOH. After removal of MeOH the precipitate was collected by centrifugation and dried in a lyophilizer to give 6.6 g of dry solid 2609-173-16 (compound 6, 3.2%).

The second product pool (4.3 g) from the C18 biotage column was processed in a similar manner to give 2.0 g of dried solid 2609-173-27 (compound 6, 3.06%). 2609-173-16 and 2609-176-27 were combined to yield 8.6 g of 2609-174-6.

2609-174-6 (400 mg) was dissolved in 1.3 mL of MeOH containing 0.1% AcOH. The solution was charged on a Phenomenex Luna C8 column and eluted with 68% MeOH / 32% water containing 0.05% AcOH at a flow rate of 24 mL / min.

Based on analytical HPLC, fractions from each run (22 runs in total) from 15.8 to 19.8 minutes were combined, evaporated to remove MeOH and lyophilized to include 2609-174-28 (12.6% Compound 6). 1.4 g). 2609-174-28 was used for final separation of compound 6 in YMC-AQ C18 column. A total of 28 runs were performed.

2609-174-28 (50 mg) was dissolved in 0.25 mL of MeOH containing 0.1% AcOH. The solution was injected into a YMC AQ C18 column. The column was eluted at 9.9 mL / min with 70% MeOH / 30% water containing 0.05% AcOH. Based on the analytical HPLC profile, generally selected fractions from 28 runs at 48.4 to 50.4 minutes were poured, evaporated and lyophilized to yield Compound 6 (2609-176-30, 85 mg).

Fractions collected immediately before 48.4 minutes and mainly comprising Compound 6 were also combined and dried to give 2609-176-35 (50 mg). Reprocessing 2609-176-35 using the same column and mobile phase (run three times) yielded another, many Compound 6, which was combined with 2609-176-30 to give about 95% chromatographic purity of the product (2609- 177-10) 102 mg. HPLC chromatograms (UV and ELSD at 205, 230 nm) and proton NMR spectra of compound 6 (2609-176-10) are shown in FIGS. 16, 17, 18 and 19, respectively. The proton NMR of 2609-176-10 is consistent with that of the standard sample of compound 6. SSI-MS of Compound 6 (FIG. 20) shows strong [M + Na] ⁺ peak at m / z 685 consistent with Formula C ₃₇ H ₅₈ O _1O of Compound 6.

Carbohydrate Removal by Enzymes

Compound 6 (10 mg) was dissolved in 5 mL of MeOH. Potassium phosphate buffer (50 mM, pH 6.0, 10 mL) was added to the MeOH solution. Cellulase (Sigma Product No. C0615, 20 mg) was dissolved in 10 ml of potassium phosphate buffer solution. Then a solution of compound 6 was added to the cellulase solution and the mixture was incubated at 37 ° C. for 3 days.

The progress of the reaction was monitored by HPLC as follows: An aliquot of the reaction mixture (20 μl) was added to MeOH (80 mL) containing 0.1% acetic acid (v / v). Samples were then analyzed on a 25 cm × 4.6 mm ID SUPELCOSIL LC-8 reverse phase HPLC column (5 μm particle size) using a 20 minute MeOH-water gradient.

After 3 days of incubation, 20 ml of CH ₂ Cl ₂ was added to the reaction mixture which was vigorously shaken and then centrifuged at 2000 rpm for 3 minutes. The aqueous layer was removed and extracted _two more times with 20 ml of CH ₂ Cl ₂ . The organic layer was combined and the solvent removed to give 9.6 mg of crude product. Crude product was dissolved in a minimum volume of 20: 1 CH ₂ Cl ₂ -MeOH and chromatographed on a silica column eluting with the same solvent mixture. Column development was performed with cerium molybdate colorant (1% CeSO _4, 5% (NH ₄ ) Mo ₇ O ₂₄ .4H ₂ O (w / v) and 10% concentrated H ₂ SO ₄ (v / v)) and 15 Monitored by TLC using a 1 CH ₂ Cl ₂ -MeOH solvent mixture. Fractions containing a substance with an Rf of 0.26 were pooled to yield compound I-16 (compound number shown in Table 1 above).

Deacetylation of Compound 6

Compound 6 (chromium purity 88%) was treated with 1% ammonia to give compound I-13 (compound number shown in Table 1 above).

Compound 6 (chromium purity 88%) was heated at 85 ° C. for 1 hour to afford compound I-13 (compound number shown in Table 1 above).

Biological assay

A. Assay to Measure the Ability of Compounds of Formula (I) to Inhibit Aβ-42

Compounds of the invention and extracts comprising the compounds can be assayed as inhibitors of amyloid-beta (1-42) peptides in vitro or in vivo. This assay is described in detail in US Pat. No. 6,649,196, the entirety of which is incorporated herein by reference.

Compounds of the invention have been found to selectively reduce amyloid-beta (1-42) peptides according to assays based on cells performed in substantially the same manner as described in US Pat. No. 6,649,196.

B. Assays to Measure the Ability of Compounds of Formula (I) to Affect the Ratio of Total Aβ

Compounds of the invention are described in Wang et al., J Biol. Chem. 1996, 50: 31894-31902, The Profile of Soluble Amyloid β Protein in Cultured Cell Media, the entirety of which is incorporated herein by reference in its entirety, using an assay protocol substantially similar to that described in 42) Assay to determine its effect on the total ratio of peptides. This assay quantified amyloid-β protein using immunoprecipitation and mass spectroscopy (IP-MS). To illustrate using compound 6, the compound was found to reduce amyloid-β (1-42) peptide, while increasing amyloid-β (1-37) peptide and amyloid-β (1-39) peptide. lost. The results are shown in FIG.

Compound 6 was also assayed according to the method described in Wang et al., In 7 W cells (APPwt) and 7PA2 cells (APP _V717F ). The APP717 mutation increased the relative amount of amyloid-β (1-42) peptide. In this assay, Compound 6 appears to reduce amyloid-β (1-42) peptide while increasing amyloid-β (1-39) peptide. The results are shown in FIG.

While various aspects of the invention have been described, it is evident that the basic examples can be modified to provide other aspects of using the compounds and methods of the invention. Therefore, it is understood that the scope of the present invention is defined by the appended claims rather than the specific aspects expressed in the manner of the embodiments.

Claims (27)

A compound of formula (I) or a pharmaceutically acceptable salt thereof. Formula I

In Formula I above, Each ring A, ring B, ring C, ring D and ring E are independently saturated or partially unsaturated aromatic rings, G is S, CH ₂ , NR or O, R ¹ and R ² are each independently halogen, R, OR, suitably protected hydroxyl group, SR, suitably protected thiol group, N (R) ₂ or suitably protected amino group, or R ¹ and R ² together form a 3 to 7 membered saturated or partially unsaturated aryl ring having 0 to 2 heteroatoms independently selected from nitrogen, oxygen and sulfur, Each R is independently hydrogen, an optionally substituted C _1-6 aliphatic group, or an optionally substituted 3-8 membered saturated or partially unsaturated having 0-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur Is an aryl ring wherein two R on the same nitrogen atom are optionally substituted with 3 to 8 membered saturated or partially unsaturated aryl rings having 1 to 4 heteroatoms independently selected from nitrogen, oxygen and sulfur Form the n is 0 to 2, R ³ , R ⁴ , R ⁷ and R ⁸ are each independently halogen, R, OR, suitably protected hydroxyl group, SR, suitably protected thiol group, SO ₂ R, OSO ₂ R, N (R) ₂ , suitably protected amino group, NR (CO) R, NR (CO) (CO) R, NR (CO) N (R) ₂ , NR (CO) OR, (CO) OR, O (CO) R , (CO) N (R) ₂ and O (CO) N (R) ₂ , m is 0 to 2, R ⁵ is TC (R ′) ₃ , TC (R ′) ₂ C (R ″) ₃ , R, OR, suitably protected hydroxyl group, SR, suitably protected thiol group, SO ₂ R, OSO ₂ R, N (R) ₂ , suitably protected amino group, NR (CO) R, NR (CO) (CO) R, NR (CO) N (R) ₂ , NR (CO) OR, (CO) OR , O (CO) R, (CO) N (R) ₂ or O (CO) N (R) ₂ , Each T is independently a valence bond, or an optionally substituted, linear or branched, saturated or unsaturated C _1-6 alkylidene chain, wherein up to two methylene units of T are optionally independently -O-,- Replaced by N (R)-, -S-, -C (O)-, -S (O)-or -S (O) _2- , Each R ′ and R ″ is independently R, OR, SR, SO ₂ R, OSO ₂ R, N (R) ₂ , NR (CO) R, NR (CO) (CO) R, NR (CO) N (R) ₂ , NR (CO) OR, (CO) OR, O (CO) R, (CO) N (R) ₂ and O (CO) N (R) ₂ , R ⁶ is halogen, R, OR, SR, SO ₂ R, OSO ₂ R, N (R) ₂ , NR (CO) R, NR (CO) (CO) R, NR (CO) N (R) ₂ , NR (CO) OR, (CO) OR, O (CO) R, (CO) N (R) ₂ or O (CO) N (R) ₂ , R ⁹ and R ^{9 '} are each independently selected from halogen, R, OR, SR and N (R) ₂ , or R ¹ and R ² together are 0 to 2 heteroatoms independently selected from nitrogen, oxygen and sulfur To form a 3-7 membered saturated or partially unsaturated aryl ring having Q is a valence bond, or an optionally substituted, linear or branched, saturated or unsaturated C _1-6 alkylidene chain, wherein up to two methylene units of Q are optionally independently -O-, -N (R) Is replaced by-, -S-, -C (O)-, -S (O)-or -S (O) _2- . The compound of claim 1, wherein G is O and R ¹ and R ² are each independently R or OR. The compound of claim 2, wherein R ¹ and R ² are each independently R, wherein R is hydrogen or an optionally substituted C _1-6 aliphatic group. 3. A compound according to claim 2, wherein R ¹ and R ² together form a 3-6 membered saturated or partially unsaturated aryl ring having 0-2 heteroatoms independently selected from nitrogen, oxygen and sulfur. The compound of claim 4, wherein R ⁵ is TC (R ′) ₃ or TC (R ′) ₂ C (R ″) ₃ , wherein each T is independently a valence bond, or a straight or branched C _1-4 alkyl As the leaden chain, one methylene unit of T is optionally replaced with -O-, -N (R)-or -S-, and each R 'and R "are independently R, OR, OC (O) R, SR or N (R) ₂ phosphorus compound. 6. The compound of claim 5, wherein Q is an optionally substituted linear or branched saturated or unsaturated C _1-2 alkylidene chain, wherein up to one methylene unit of Q is optionally -O-, -N (R). Or a compound replaced with -S-. The compound of claim 6, wherein Q is —O—. The compound of claim 1, wherein the compound is a compound of Formula Vc or a pharmaceutically acceptable salt thereof. Formula Vc

The compound of claim 1, wherein the compound is a compound of Formula Vd or Ve or a pharmaceutically acceptable salt thereof. Chemical Formula Vd

Chemical Formula Ve

The compound of claim 1, wherein the compound is a compound of Formula Vf or a pharmaceutically acceptable salt thereof. Formula Vf

The compound of claim 10, wherein the compound is a compound of Formula Vg or Vh or a pharmaceutically acceptable salt thereof. Formula Vg

Formula Vh

12. The compound of claim 11, wherein R ¹ and R ² together form a 3-6 membered saturated carbocyclic ring, wherein R ⁷ is -OH. The compound of claim 1, wherein the compound is selected from compounds of formula VII, VIII, IX or X. Formula VII

Formula VIII

Formula IX

Formula X

Compound selected from. The compound of claim 14, wherein said compound is

Compound selected from. (A) providing a compound of formula (Vb) or a pharmaceutically acceptable salt thereof; and A process for preparing a compound of formula Va or a pharmaceutically acceptable salt thereof, comprising the step (b) of treating a compound of formula Vb with a suitable enzyme to form a compound of formula Va. Chemical formula Va

Formula Vb

In the above formulas Va and Vb, Each ring A, ring B, ring C, ring D and ring E are independently saturated or partially unsaturated aromatic rings, G is S, CH ₂ , NR or O, R ¹ and R ² are each independently halogen, R, OR, suitably protected hydroxyl group, SR, suitably protected thiol group, N (R) ₂ or suitably protected amino group, or R ¹ and R ² together form a 3 to 7 membered saturated or partially unsaturated aryl ring having 0 to 2 heteroatoms independently selected from nitrogen, oxygen and sulfur, Each R is independently hydrogen, an optionally substituted C _1-6 aliphatic group, or an optionally substituted 3-8 membered saturated or partially unsaturated having 0-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur Is an aryl ring wherein two R on the same nitrogen atom are optionally substituted with 3 to 8 membered saturated or partially unsaturated aryl having 1 to 4 heteroatoms independently selected from nitrogen, oxygen and sulfur, together with said nitrogen atom; Forming a ring, n is 0 to 2, R ³ , R ⁴ , R ⁷ and R ⁸ are each independently halogen, R, OR, suitably protected hydroxyl group, SR, suitably protected thiol group, SO ₂ R, OSO ₂ R, N (R) ₂ , suitably protected amino group, NR (CO) R, NR (CO) (CO) R, NR (CO) N (R) ₂ , NR (CO) OR, (CO) OR, O (CO) R , (CO) N (R) ₂ and O (CO) N (R) ₂ , m is 0 to 2, R ⁵ is TC (R ′) ₃ , TC (R ′) ₂ C (R ″) ₃ , R, OR, suitably protected hydroxyl group, SR, suitably protected thiol group, SO ₂ R, OSO ₂ R, N (R) ₂ , suitably protected amino group, NR (CO) R, NR (CO) (CO) R, NR (CO) N (R) ₂ , NR (CO) OR, (CO) OR , O (CO) R, (CO) N (R) ₂ or O (CO) N (R) ₂ , Each T is independently a valence bond, or an optionally substituted, linear or branched, saturated or unsaturated C _1-6 alkylidene chain, wherein up to two methylene units of T are optionally independently -O-,- Replaced with N (R)-, -S-, -C (O)-, -S (O)-or -S (O) _2- , Each R ′ and R ″ is independently R, OR, SR, SO ₂ R, OSO ₂ R, N (R) ₂ , NR (CO) R, NR (CO) (CO) R, NR (CO) N (R) ₂ , NR (CO) OR, (CO) OR, O (CO) R, (CO) N (R) ₂ or O (CO) N (R) ₂ , R ⁶ is halogen, R, OR, SR, SO ₂ R, OSO ₂ R, N (R) ₂ , NR (CO) R, NR (CO) (CO) R, NR (CO) N (R) ₂ , NR (CO) OR, (CO) OR, O (CO) R, (CO) N (R) ₂ or O (CO) N (R) ₂ , R ⁹ and R ^{9 '} are each independently selected from halogen, R, OR, SR and N (R) ₂ , or R ¹ and R ² together are 0 to 2 heteroatoms independently selected from nitrogen, oxygen and sulfur To form a 3-7 membered saturated or partially unsaturated aryl ring having R ¹⁰ is a sugar-containing or sugar-like residue. 17. The method of claim 16, wherein said suitable enzyme is cellulase, xylanase, xylosidase, glycyrrhizin acid hydrolase or glucuronidase. A composition comprising a compound according to claim 1 and a pharmaceutically acceptable carrier, adjuvant or vehicle. A method of inhibiting the production of amyloid-beta peptides in a patient, comprising administering the composition of claim 18 to the patient. A method of inhibiting the production of amyloid-beta (1-42) peptides in a patient, comprising administering the composition of claim 18 to the patient. 20. The method of claim 19, wherein the method does not affect the Notch process. The method of claim 21, wherein the amyloid-beta (1-42) peptide level is reduced and the amyloid-beta (1-40) peptide level is not substantially reduced. The method of claim 22, wherein the level of one or more of amyloid-beta (1-37) and amyloid-beta (1-39) is increased. A method of treating or lessening the severity of a disorder associated with an amyloid-beta (1-42) peptide, comprising administering a composition according to claim 18 to a patient. The method of claim 24, wherein the disorder is Alzheimer's disease, Parkinson's disease, Down syndrome, inclusion body myositis, cerebral amyloid angiopathy, mild cognitive impairment, Lewy body dementia, cataract, tau disease, Huntington's disease, muscular dystrophy ( ALS / Lou Gehrig's disease), type 2 diabetes, Transthyretin amyloid disease or prion diseases (Creutzfeldt-Jakob disease, Gerstmann-Steuisler-Shinker syndrome, fatal familial insomnia and kuru) ]. The method of claim 25, wherein the disorder is Alzheimer's disease, Parkinson's disease or Down syndrome. A method of reducing amyloid-beta (1-42) peptide levels in a cell, comprising contacting the compound of claim 1 with the cell.

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