HK1190717B

HK1190717B - (-)- huperzine a processes and related compositions and methods of treatment

Info

Publication number: HK1190717B
Application number: HK14103893.6A
Authority: HK
Inventors: S．赫尔森; M．K．M．邓
Original assignee: 耶鲁大学
Priority date: 2011-03-04
Filing date: 2012-02-17
Publication date: 2017-11-24

Description

(-) -huperzine A methods and related compositions and methods of treatment

Related applications/research support

This application claims priority to U.S. provisional application serial No. 61/449,198 entitled "Huperzine a," filed on 3, month 4, 2011, the entire contents of which are incorporated herein by reference.

Background

(-) -huperzine A (1) is a tricyclic alkaloid derived from Huperzia serrata (Huperzia serrata).¹(-) -huperzine A (1) is a potent, selective and reversible inhibitor of acetylcholinesterase (AChE, Ki =23 nM).²Recent studies have established that this activity can be exploited against organophosphate chemical warfare agents such as sarin and VX by inhibiting their covalent modification of peripheral and brain AChE。³There is also substantial evidence that (-) -huperzine a (1) slows the progression of neurodegenerative diseases, including alzheimer's disease.⁴(-) -huperzine A (1) is well tolerated in humans even at doses well above those clinically required.⁵Therefore, clinical studies of (-) -huperzine A (1) are the subject of intensive research in the pharmaceutical and defense industries.

One of the major obstacles to clinical development of (-) -huperzine a (1) is supply. The yields from natural sources are low (average yield of dry herbs = 0.011%),^4aand over-harvesting has resulted in a rapid reduction in huperzia serrata (Huperziaceae) resources.⁶Combining these problems, production species take nearly 20 years to mature.⁶

The total synthesis provides an alternative potential source of huperzine. Enantioselective synthesis is highly desirable because (+) -huperzine A is significantly less potent than the natural (-) -enantiomer (1).⁷The earliest total synthesis of (+ -) -huperzine A was performed by Kozikowski and Xia⁸And Qian and Ji.⁹It was reported. Subsequently, Kozikowski et al.¹⁰Routes based on chiral auxiliaries were developed. In the years that follow, various groups have reported improvements to the Kozikowski route,¹¹and the integrity of huperzine,¹²Part of the material,¹³And formally¹⁴And (4) routing. Nevertheless, 16 steps are required and a yield of about 2.8% of a route based on Kozikowski chiral control agent¹⁰It remains the most efficient published route for the synthesis of (-) -huperzine A (1).¹⁵

In view of the large number of steps and relatively low stereochemical yields of the known processes for the preparation of (-) -huperzine a, and the growing importance of (-) -huperzine a as a neuroprotective agent, there is a need for improved processes for the preparation of substantially pure (-) -huperzine a in yields that facilitate scaling up to commercial manufacture.

Disclosure of Invention

Summary of The Invention

In one embodiment, the present invention provides a novel method for preparing substantially pure (-) huperzine a and substantially pure (-) huperzine a derivatives in relatively high yield by synthesis employing significantly fewer steps than known techniques.

In another embodiment, the present invention provides a novel process for the preparation of various intermediates useful in the manufacture of pharmaceutically active ingredients, including substantially pure (-) huperzine a and substantially pure (-) huperzine a derivatives.

In yet another embodiment, the present invention provides various novel compositions useful in the manufacture of pharmaceutically active ingredients, including substantially pure (-) huperzine a and substantially pure (-) huperzine a derivatives.

In yet another embodiment, the present invention provides a method of treating or preventing a neurological disorder, comprising administering substantially pure (-) huperzine a or a substantially pure (-) huperzine a derivative to a subject having or at risk of developing a neurological disorder.

In yet another embodiment, the present invention provides a novel process for preparing substantially pure (-) huperzine A having the formula:

comprising reacting an amide of the formula:

subjecting to a modified Hoffmann reaction (Hoffmann reaction) in an aqueous or alcoholic solvent, preferably methanol, and in the presence of bis (trifluoroacetoxyiodo) benzene (PIFA) to form an intermediate, deprotecting the intermediate completely to form (-) huperzine a, and purifying the (-) huperzine a (e.g., by crystallization and/or flash column chromatography) to obtain substantially pure (-) huperzine a.

As used herein, "substantially pure (-) huperzine a" includes greater than about 80% by weight (-) huperzine a and less than about 20% by weight (+) huperzine a, more preferably greater than about 90% by weight (-) huperzine a and less than about 10% by weight (+) huperzine a, even more preferably greater than about 95% by weight (-) huperzine a and less than about 5% by weight (+) huperzine a, and most preferably greater than about 99% by weight (-) huperzine a and less than about 1% by weight (+) huperzine a. The nearly pure (-) huperzine a derivative contains more than 99.5% by weight (-) huperzine a and less than 0.5% by weight (+) huperzine a, more preferably more than about 99.9% (-) huperzine a and less than about 0.1% (+) huperzine a. "substantially pure (+) huperzine a derivative" is defined similarly with respect to the relative amounts of the (+) and (-) enantiomers thereof.

As used herein, the term (±) huperzine A (or "racemic huperzine A" or "huperzine A racemate") means a composition comprising about 40-60% of (-) huperzine A and about 40-60% of (+) huperzine A. The racemate of the huperzine a derivative is defined similarly for its (-) and (+) enantiomers.

"huperzine a derivative" (e.g., as used in the term "substantially pure (-) huperzine a derivative") refers to compounds as described in U.S. patent No. RE38460, as well as compounds of formula (II) and (III) described below.

In one embodiment, the amide subjected to the modified hofmann reaction as described above is preferably prepared in a one-pot process by a process comprising the steps of: reacting a cyanohydrin of the formula:

dehydrating in an organic solvent, preferably toluene, under heating and in the presence of a Burgess reagent (Burgessrean) to form a dehydration product, and subjecting the dehydration product to pyrolysis in an alcohol, preferably aqueous ethanol, in the presence of a platinum catalyst to form an amide. This novel reaction also constitutes an embodiment of the invention and can also be carried out stepwise.

In one embodiment, the above cyanohydrins are preferably prepared in a one-pot process by: reacting an olefination product substantially in the form of the E isomer and having the formula:

undergo oxidative desilylation (e.g., by reaction with boron trifluoride-acetic acid complex or a Bronsted acid such as TFA, MSA, FMSA, or tetrafluoroboric acid in an inert solvent such as DCM, or by oxidation using a Fleming-Tamao, followed by fluoride, hydrogen peroxide, and potassium carbonate). In addition to the protic acid, removal of the silyl group involves a step of treatment with fluoride, hydrogen peroxide and potassium carbonate. This new reaction step also constitutes an embodiment of the present invention.

In one embodiment, the above-described alkylene products are preferably prepared in a one-pot process in a process comprising deprotonating an addition-alkylation product of the formula:

reacting the addition-alkylation product with lithium bis (trimethylsilyl) amide (LHMDS) or Lithium Diisopropylamide (LDA) and an electrophilic cyanide source (e.g., p-toluenesulfonylcyanide, cyanogenbromide, etc.) in an organic solvent (e.g., THF or toluene) to form an alpha-cyanoketone, subjecting the alpha-cyanoketone to palladium-catalyzed (e.g., tetrakis (triphenylphosphine) palladium, tris (dibenzylideneacetone) dipalladium, palladium bis (tri-tert-butylphosphine)) intramolecular enolate heteroarylation in the presence of a base (most preferably sodium tert-butoxide) and a palladium catalyst to form a cyclization product, and in the presence of a base (e.g., n-butyllithium, sodium bis (trimethylsilyl) amide, lithium bis (trimethylsilyl) amide, potassium bis (trimethylsilyl) amide, or lithium diisopropylamide) and in an organic solvent (e.g., THF, Diethyl ether or 1, 4-dioxane) to form the olefination product, wherein the stereoselective olefination of the cyclized product kinetically favors the formation of the olefination product as the E-isomer. This new reaction step also constitutes an embodiment of the invention and can also be carried out stepwise.

In one embodiment, the addition-alkylation product is prepared in a one-pot process in a process comprising the steps of: reacting (R) -4-methyl-cyclohex-2-en-1-one with lithium dimethylphenylsilylcuprate in a conjugate addition reaction to form an initial enolate and alkylating the initial enolate by 3-bromo-2- (bromomethyl) -6-methoxypyridine) to form an addition-alkylated product. This new reaction step also constitutes an embodiment of the invention and can also be carried out stepwise.

In yet another embodiment, the present invention provides a method of cyclizing a β -keto comprising subjecting an α -cyanoketone to palladium-catalyzed intramolecular enolate heteroarylation, as described in detail below.

In yet another embodiment, the present invention provides a novel process for preparing substantially pure (-) huperzine a comprising:

(a) preferably in a one-pot process, (R) -4-methyl-cyclohex-2-en-1-one is reacted with lithium dimethylphenylsilylcuprate in a conjugate addition reaction to form an initial enolate and the initial enolate is alkylated by 3-bromo-2- (bromomethyl) -6-methoxypyridine) to form an addition-alkylation product having the formula:

(b) the addition-alkylation product is deprotonated, preferably in a one-pot process, by: reacting the addition-alkylation product with lithium bis (trimethylsilyl) amide (LHMDS) or Lithium Diisopropylamide (LDA) in an organic solvent (e.g., THF or toluene) to form an alpha-cyanoketone, subjecting the alpha-cyanoketone to palladium-catalyzed intramolecular enolate heteroarylation in the presence of a base (most preferably sodium tert-butoxide) to form a cyclized product, and stereoselective olefination of the ketone function of the cyclized product in the presence of a base (e.g., sodium n-butyllithium, bistrimethylsilyl) amide lithium, bistrimethylsilyl) amide potassium or lithium diisopropylamide) and in an organic solvent (e.g., THF, diethyl ether or 1, 4-dioxane) in a Wittig olefination reaction to form an olefination product, wherein the stereoselectivity of the cyclization product kinetically favors the formation of the olefination product as an E-isomer and wherein the olefination product has the property of being an E The following formula:

(c) the alkylene product is subjected to oxydimethylsilylation (e.g., by reaction with boron trifluoride-acetic acid complex or a bronsted acid such as TFA, MSA, FMSA, or tetrafluoroboric acid in an inert solvent such as DCM, or by oxidation using a fleming-jade tail) to form a cyanohydrin of the formula:

(d) preferably in a one-pot process, cyanohydrin in an organic solvent (preferably toluene) is dehydrated under heating and in the presence of a burges reagent to form a dehydration product, and the dehydration product is subjected to pyrolysis in an alcohol (preferably ethanol) and in the presence of a platinum catalyst to form an amide having the formula:

and

(f) subjecting an amide to a modified hofmann reaction in an aqueous or alcoholic solvent, preferably methanol, and in the presence of bis (trifluoroacetoxyiodo) benzene (PIFA) to form an intermediate, fully deprotecting the intermediate to form (±) huperzine a, and purifying the (±) huperzine a (e.g. by flash column chromatography) to obtain substantially pure (-) huperzine a:

in yet another embodiment, the invention provides a compound of formula (I):

wherein:

R₁selected from substituted or unsubstituted C₁-C₆Alkyl and substituted or unsubstituted ether;

R₂and R₅Independently selected from H, substituted or unsubstituted C₁-C₆Alkyl and CN, with the proviso that when R is₂Or R₅When one is CN, the other must be H;

x is halogen;

R₃independently at each occurrence, selected from H, substituted or unsubstituted C₁-C₆Alkyl, ether, amino and alkoxy groups;

R₄selected from Si (CH)₃)₂Ph, substituted or unsubstituted C₁-C₆Alkyl, substituted or unsubstituted C₁-C₆Alkenyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl;

a is C, N or S;

m is 0, 1 or 2;

n is 0 or 1;

or a pharmaceutically acceptable salt, enantiomer, diastereomer, solvate or polymorph thereof.

In yet another embodiment, the present invention provides a compound of formula (II):

wherein:

R₂and R₅Independently selected from H and substituted or unsubstituted C₁-C₆An alkyl group;

R₄selected from H, OH and Si (CH)₃)₂Ph；

R₆Is selected from NH₂Amides, CN, carboxylic acid derivatives (e.g. esters, ketones, or secondary or tertiary amines), alcohols or aldehydes;

R₇is substituted or unsubstituted C₁-C₆An alkyl, ester, or substituted or unsubstituted aryl;

a is C, N or S; and is

n is 0 or 1;

In one embodiment, the compounds of formula (I) and (II) are used to prepare pharmacologically active compositions comprising substantially pure (-) huperzine A and substantially pure (-) huperzine A derivatives.

Preferred compounds of the invention include:

wherein R is¹And R²Each independently is H or C₁-C₆An alkyl group;

and primary amine derivatives thereof (wherein CN is converted to CH)₂NR¹R²Group, wherein R¹And R²The same as described above);

and primary amine derivatives thereof (wherein CN is converted to CH)₂NR¹R²Group, wherein R¹And R²The same as described above); and

In yet another embodiment, the present invention provides a novel process for preparing substantially pure (-) huperzine a or a derivative thereof having the formula (III):

wherein:

R₄selected from H, OH and Si (CH)₃)₂Ph；

a is C, N or S; and is

n is 0 or 1;

comprising reacting an amide having the following formula (IV):

wherein R is₁、R₂、R₃、R₄、R₅、R₇A and n are defined as compounds of formula (III), are subjected to an improved Hofmann reaction in an aqueous or alcoholic solvent, preferably methanol, and in the presence of bis (trifluoroacetoxyiodo) benzene (PIFA) to form an intermediate, the intermediate is fully deprotected to form (±) huperzine A or (±) huperzine A derivatives, and the (±) huperzine A or (±) huperzine A derivatives are purified, for example by flash column chromatography, to obtain substantially pure (-) huperzine A or substantially pure (±) huperzine A derivatives.

In yet another embodiment, the present invention provides a process for preparing an amide having formula (IV):

wherein:

R₄selected from H, OH and Si (CH)₃)₂Ph and H;

a is C, N or S; and is

n is 0 or 1;

comprising reacting a cyanohydrin of formula (V):

wherein R is₁、R₂、R₃、R₅、R₇A and n are as defined for the compound of formula (IV), dehydrating in an organic solvent, preferably toluene, with heating and in the presence of a Burgis reagent to form a dehydration product, and subjecting the dehydration product to pyrolysis in an alcohol, preferably ethanol, in the presence of a platinum catalyst to form an amide, wherein the dehydrating and the pyrolysis can be carried out in one-pot or in steps.

In yet another embodiment, the present invention provides a process for preparing cyanohydrins of formula (V):

wherein:

a is C, N or S; and is

n is 0 or 1;

comprising reacting an olefination product substantially in the form of the E isomer and having formula (VI):

wherein R is₁、R₂、R₃、R₅、R₇A and n are as defined for compounds of formula (V), are subjected to oxydimethylsilylation (e.g., by reaction with boron trifluoride-acetic acid complex or a bronsted acid such as TFA, MSA, FMSA or tetrafluoroboric acid in an inert solvent such as DCM, or by oxidation using fleming-jade tail), wherein the process can be carried out in a one-pot process or in steps.

In another embodiment, the present invention provides a process for preparing an olefination product substantially in the E isomeric form and having the formula (VI):

wherein:

a is C, N or S; and is

n is 0 or 1;

comprising reacting an addition-alkylation product having the formula (VII):

wherein R is₁、R₂、R₃、R₅A and n are as defined in (V), R₄Selected from Si (CH)₃)₂Ph, substituted or unsubstituted C₁-C₆Alkyl, substituted or unsubstituted C₁-C₆Alkenyl, substituted or unsubstituted aryl and substituted or unsubstituted heteroaryl, X is halogen and m is 0, 1 or 2, by reacting the addition-alkylation product with lithium bis (trimethylsilyl) amide (LHMDS) or Lithium Diisopropylamide (LDA) in an organic solvent (e.g., THF or toluene) to form α -cyanoketone, subjecting α -cyanoketone in the presence of a base (most preferably sodium tert-butoxide) to deprotonationSubjecting the cyclized product to palladium-catalyzed intramolecular enolate heteroarylation to form a cyclized product, and stereoselective olefination of the ketone function of the cyclized product in the presence of a base (e.g., sodium n-butyllithium, bistrimethylsilyl) amide lithium, bistrimethylsilyl) amide potassium or lithium diisopropylamide and in an organic solvent (e.g., THF, diethyl ether, or 1, 4-dioxane) in a Wittig olefination reaction to form an olefination product, wherein each of the foregoing reactions can be carried out in a one-pot or in steps.

These and other aspects of the invention will be further detailed in the detailed description.

Drawings

FIG. 1 shows a comparison of NMR data for synthetic and natural (-) -huperzine A.

Figure 2 includes a list of nmr and ir spectra of the compositions of the invention.

FIG. 3 shows that the minor diastereomer of the olefinating product prepared according to the process of the present invention was confirmed to have the Z-configuration by NOE analysis (500MHz, CDCl 3).

Detailed Description

The following terminology is used to describe the invention, among other things. It is to be understood that terms not specifically defined will have meanings consistent with their usage in the context of the present invention as understood by those of ordinary skill in the art.

The term "compound" as used herein, unless otherwise indicated, refers to any specific compound disclosed herein, and includes tautomers, regioisomers, geometric isomers, and where applicable, optical isomers (e.g., enantiomers), stereoisomers (diastereomers), and pharmaceutically acceptable salts and derivatives thereof (including prodrug forms). The term "compound" when used in this context generally refers to a single compound, but may also include other compounds such as stereoisomers, regioisomers and/or optical isomers (including racemic mixtures) as well as specific enantiomers or enantiomerically enriched mixtures and diastereomers and epimers (as the context applies) of the disclosed compounds. The term also refers in this context to prodrug forms of the compounds that are modified to facilitate administration and delivery of the compounds to the active site.

The term "patient" or "subject" is used throughout the context of the present specification to describe an animal, typically a mammal and preferably a human, to which treatment (including prophylactic treatment, i.e. prevention) is provided by a composition according to the invention. For the treatment of those particular infections, conditions or diseases with respect to a particular animal, e.g., a human patient, the term "patient" refers to that particular animal.

The symbols used in the compounds according to the inventionTo indicate that the bond between atoms is a single or double bond depending on the context in which the bond is used in the compound, depending on the atom (and substituent) used to define the compound. Thus, when a carbon (or other) atom is used and the context in which the atom is used requires a double or single bond to connect the atom to an adjacent atom in order for the atom used to maintain the proper valence, then the bond is considered a double or single bond.

"neurological disorders" include, but are not limited to, amyloid-related disorders such as alzheimer's disease and the amyloid disorders described below; psychiatric disorders such as Gilles de la Tourette syndrome, Post Traumatic Stress Disorder (PTSD), panic and anxiety disorders, obsessive compulsive disorder and schizophrenia; developmental disorders such as fragile X syndrome and autism; pain; drug addiction, such as alcohol abuse; neurodegenerative diseases such as parkinson's disease and huntington's chorea; and stroke and ischemic brain injury, amyotrophic lateral sclerosis and epilepsy. "neurological disorder" also includes any disorder, symptom or effect associated with or associated with exposure to a neurotoxin, including but not limited to a neurotoxin such as a chemical warfare agent.

"amyloid-related disorders" include diseases associated with amyloid accumulation that may be localized to one organ ("localized amyloidosis") or spread to multiple organs ("systemic amyloidosis"). Secondary amyloidosis may be associated with chronic infection (such as tuberculosis) or chronic inflammation (such as rheumatoid arthritis), including secondary amyloidosis, also seen in the familial form of Familial Mediterranean Fever (FMF), and another type of systemic amyloidosis present in long-term hemodialysis patients. Localized forms of amyloidosis include, but are not limited to, type II diabetes and any related disorders thereof, neurodegenerative diseases such as itch, bovine spongiform encephalopathy, Creutzfeldt-Jakob disease, alzheimer's disease, Senile Systemic Amyloidosis (SSA), cerebral amyloid angiopathy, parkinson's disease, and prion protein related disorders such as prion-related encephalopathy, and rheumatoid arthritis.

Unless otherwise indicated, the term "effective" is used herein to describe an amount of a compound or composition that is used in the context to produce or achieve an intended result, whether the result relates to inhibition of the effect of the neurological disorder or to enhancing the effect of a supplemental treatment used to treat the neurological disorder (e.g., an antipsychotic or as described elsewhere herein). The term includes all other effective amount or effective concentration terms (including the term "therapeutically effective") described elsewhere in this application.

The term "treating" (treating, etc.) as used herein refers to any measure that provides a beneficial effect to a patient at risk of developing or afflicted with a neurological disorder, including alleviating or inhibiting at least one symptom of the neurological disorder, delaying the progression of the neurological disorder, or reducing the likelihood of developing the neurological disorder. "treatment" as used herein encompasses both prophylactic and therapeutic treatment.

The term "pharmaceutically acceptable salt" or "salt" is used throughout the specification to describe salt forms of one or more of the compositions herein, either to increase the solubility of the compound in saline for parenteral delivery or to increase the solubility of the compound in the gastric juices of the gastrointestinal tract of a patient in order to facilitate dissolution and bioavailability of the compound. Pharmaceutically acceptable salts include those derived from pharmaceutically acceptable inorganic or organic bases and acids. Suitable salts include those derived from alkali metals (such as potassium and sodium), alkaline earth metals (such as calcium, magnesium) and ammonium salts among many other acids well known in the pharmaceutical art. As the composition containing a neutralized salt of a carboxylic acid and free phosphoric acid according to the present invention, sodium salts and potassium salts may be preferable. The term "salt" shall mean any salt consistent with the use of the compounds according to the present invention. Where the compound is used for a pharmaceutical indication, the term "salt" shall mean a pharmaceutically acceptable salt consistent with the use of the compound as a medicament.

The term "co-administration" shall mean that at least two compounds or compositions are administered to a patient simultaneously such that an effective amount or concentration of each of the two or more compounds may be present in the patient at a given point in time. While the compounds according to the invention may be co-administered to a patient simultaneously, the term encompasses the administration of two or more agents together at the same time or at different times, including sequential administration. Preferably, an effective concentration of all co-administered compounds or compositions is present in the subject at a given time.

For example, a compound according to the present invention may be administered with one or more agents useful for treating an amyloid-related disorder or a stage of an amyloid-related disorder. The type of co-administered agent may vary widely depending on the particular clinical setting. For example, co-administered agents may include anticoagulants or coagulation inhibitors, antiplatelet or platelet inhibitors, thrombin inhibitors, thrombolytics or fibrinolytics, antiarrhythmics, antihypertensives, calcium channel blockers (L-and T-type), cardiac glycosides, diuretics, mineralocorticoid receptor antagonists, phosphodiesterase inhibitors, cholesterol/blood fat lowering agents and lipid profile therapies (lipid profile therapies), antidiabetic agents, antidepressants, anti-inflammatory agents (steroids and non-steroids), antiosteoporotic agents, hormone replacement therapies, oral contraceptives, antiobesity agents, anxiolytic agents, antiproliferative agents, antineoplastic agents, antiulcer and gastroesophageal reflux disease agents, growth hormone and/or growth hormone secretagogues, thyroid mimetics (including thyroid receptor antagonists), anti-infective agents, antiviral agents, anti-viral agents, anti-inflammatory agents, Antibacterial and antifungal agents.

More specifically, in the case of Alzheimer's disease, additional agents that may be used include, but are not limited to, cholinesterase inhibitors, the antioxidant Ginkgo biloba extract, non-steroidal anti-inflammatory agents, and non-specific NMDA antagonists, such as(memantine) for parkinson's disease, additional agents that may be used include, but are not limited to carbidopa/levodopa (Sinemet-Bristol Myers Squibb) other therapies including dopamine agonists, carbidopa/levodopa therapies, COMT inhibitors, anticholinergics, and MAO inhibitors, such as selegiline/propynedrine for type II diabetes, additional agents that may be used include, but are not limited to biguanides (e.g., metformin), glucosidase inhibitors (e.g., acarbose), insulin (including insulin secretagogues or insulin sensitizers), meglitic acids (e.g., repaglinide), sulfonylureas (e.g., glimepiride, glyburide, and glipizide), biguanide/glibenclamide complexes (e.g., glucovence), thiazolidinediones (e.g., troglitazone, rosiglitazone and pioglitazone), PPAR- α agonists, PPAR- γ agonists, PPAR 42/γ agonists, dual agonists, inhibitors of fatty acid binding to GLP 1-DP 36, peptidyl peptidase IV (sgp 36), and peptidyl peptidase IV).

The terms "antagonist" and "inhibitor" are used interchangeably to refer to agents specifically disclosed herein that reduce or inhibit a biological activity, such as inhibiting the activity of a neurological disorder, including, inter alia, chemical agents. A "modulator of a neurological disorder" inhibits or enhances the activity of the neurological disorder.

The term "acyl" is art-recognized and refers to a group represented by the general formula hydrocarbyl C (O) -preferably alkyl C (O) -group.

The term "amido" is art-recognized and refers to a moiety having an amino group and an acyl group and may include the same substituents as disclosed elsewhere herein.

The term "aliphatic group" refers to a straight, branched or cyclic aliphatic hydrocarbon group and includes saturated and unsaturated aliphatic groups such as alkyl, alkenyl and alkynyl groups.

The term "alkenyl" as used herein refers to an aliphatic group containing at least one double bond and is intended to include both "unsubstituted alkenyls" and "substituted alkenyls," the latter of which refers to alkenyl moieties having substituents replacing a hydrogen on one or more carbons of the alkenyl. Such substituents may be present on one or more carbons that may or may not be present in one or more double bonds. Further, such substituents may include all those contemplated for alkyl groups as discussed herein, except when the stability of the moiety is inhibited. For example, substitution of alkenyl groups by one or more alkyl, carbocyclyl, aryl, heterocyclyl or heteroaryl groups is contemplated.

The term "alkoxy" (alkoxyl) as used herein refers to an alkyl group as defined below having an oxygen radical attached thereto. Representative alkoxy groups include methoxy, ethoxy, propoxy, t-butoxy, and the like.

An "ether" is two hydrocarbons covalently linked by oxygen. Thus, the substituent of an alkyl group which renders the alkyl group an ether is an alkoxy group or is analogous to an alkoxy group, such as may be represented by- -O-alkyl, - -O-alkenyl, - -O-alkynyl, - -O- - (CH)₂)_m-one of the substituents represents, wherein m is 0 to 6, and the substituent is aryl or substituted aryl, cycloalkyl, cycloalkenyl, heterocycle or polycycle (two or three rings), each of which may be optionally substituted.

The term "alkyl" refers to a radical of a saturated aliphatic group,including straight chain alkyl, branched chain alkyl, cycloalkyl (alicyclic hydrocarbon groups), alkyl-substituted cycloalkyl and cycloalkyl-substituted alkyl groups. In a preferred embodiment, the linear or branched alkyl group has 10 or less carbon atoms in its main chain (e.g., linear C)₁-C₁₀C of a branched chain₁-C₁₀) More preferably 8 or less, most preferably 6 or less. Likewise, preferred cycloalkyl groups have 3 to 10 carbon atoms in their ring structure, more preferably 5,6,7 or 8 carbons in the ring structure.

Furthermore, the term "alkyl" (or "lower alkyl") as used throughout the specification, examples and claims is intended to include both "unsubstituted alkyls" and "substituted alkyls," the latter of which refers to alkyl moieties having substituents replacing a hydrogen on one or more carbons of the hydrocarbon backbone. Such substituents include, for example, halogen, hydroxyl, carbonyl (such as carboxyl, alkoxycarbonyl, formyl, or acyl), thiocarbonyl (such as thioester, thioacetate, or thioformate), alkoxy, phosphoryl, phosphate, phosphonate, phosphinate, amino, amido, amidine, imine, cyano, nitro, azido, sulfhydryl, alkylthio, sulfate, sulfonate, sulfamoyl, sulfonamide, sulfonyl, heterocyclyl, aralkyl, or an aromatic or heteroaromatic moiety or as described elsewhere herein. Those skilled in the art will appreciate that each substituent chemical moiety may itself be substituted. For example, the substituents of a substituted alkyl group can include substituted and unsubstituted forms of the following groups: amino, azido, imino, amido, phosphoryl (including phosphate and phosphonate), sulfonyl (including sulfate, sulfonamide, sulfamoyl and sulfonate) and silyl groups, as well as ether, alkylthio, carbonyl (including ketone, aldehyde, carboxylate and ester), -CF₃- -CN, etc. Exemplary, non-limiting, substituted alkyls are described herein. Cycloalkyl may be further substituted by alkyl, alkenyl, alkynyl, alkoxy, alkylthio, aminoalkyl, carbonyl, - -CF₃-CN, etc.

Similar substitutions may be made to alkenyl and alkynyl groups to produce, for example, but not limited to, aminoalkenyl, aminoalkynyl, amidoalkenyl, amidoalkynyl, iminoalkenyl, iminoalkynyl, thioalkenyl, thioalkynyl, carbonyl-substituted alkenyl or alkynyl groups.

As used herein, "lower alkyl" means an alkyl group as defined above but having from one to eight carbons, more preferably from one to six carbon atoms in its backbone structure, unless the number of carbons is otherwise specified. Likewise, "lower alkenyl" and "lower alkynyl" have similar chain structures. Throughout this application, preferred alkyl groups are lower alkyl groups. In a preferred embodiment, the substituents specified herein are lower alkyl.

The term "alkynyl" as used herein refers to an aliphatic group containing at least one triple bond and is intended to include both "unsubstituted alkynyls" and "substituted alkynyls," the latter of which refers to alkynyl moieties having substituents replacing a hydrogen on one or more carbons of the alkynyl group. Such substituents may be present on one or more carbons that may or may not be contained in one or more triple bonds. Further, such substituents may include all those contemplated for alkyl groups as discussed above, except when stability is inhibited. For example, substitution of alkynyl groups by one or more alkyl, carbocyclyl, aryl, heterocyclyl or heteroaryl groups is contemplated.

The term "alkylthio" refers to an alkyl group as defined above having a sulfur radical attached thereto. In a preferred embodiment, "alkylthio" is through-S-alkyl, -S-alkenyl, -S-alkynyl and-S- (CH)₂)_mOne of the substituents represents, wherein m is 0 or an integer from 1 to 8 and the substituent is as defined herein and elsewhere (R of amine/amino group) below₉And R₁₀) The same is true. Representative alkylthio groups include methylthio, ethylthio, and the like.

The terms "amine" and "amino" are art-recognized and refer to both unsubstituted and substituted amines, such as moieties that can be represented by, but are not limited to, the following general formula:

wherein R is₉、R₁₀And R'₁₀Each independently represents hydrogen, alkyl, alkenyl, - - (CH)₂)_m--R₈Or R is₉And R₁₀Taken together with the N atom to which they are attached to form a heterocyclic ring having from 4 to 8 atoms in the ring structure; r₈Represents aryl, cycloalkyl, cycloalkenyl, heterocycle or polycycle; and m is zero or an integer in the range of 1 to 8. In a preferred embodiment, R₉Or R₁₀Only one may be a carbonyl group, e.g. R₉、R₁₀Together with nitrogen, do not form an imide. In certain such embodiments, R₉And R₁₀Are not linked to N via a carbonyl group, e.g. the amine is not an amide or an imide, and the amine is preferably basic, e.g. its conjugate acid has a pK higher than 7_a. In an even more preferred embodiment, R₉And R₁₀(and optionally R'₁₀) Each independently represents hydrogen, alkyl, alkenyl or- - (CH)₂)_m--R₈. Thus, the term "alkylamine" as used herein means an amine group as defined above having a substituted or unsubstituted alkyl group attached thereto, i.e., R₉And R₁₀At least one of (a) and (b) is an alkyl group. Each group bonded to an amine group may be optionally substituted where applicable.

The term "amido" is art-recognized as an amino-substituted carbonyl and includes moieties that can be represented by the general formula:

wherein R is₉、R₁₀As defined above. Preferred embodiments of the amides will not include imides that may be unstable.

The term "aralkyl" as used herein refers to an alkyl group substituted with an aryl group (e.g., an aromatic or heteroaromatic group).

The term "aryl" as used herein includes 5,6 and 7 membered monocyclic or aromatic groups containing from zero to four heteroatoms, such as benzene, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, triazole, pyrazole, pyridine, pyrazine, pyridazine and pyrimidine, and the like. Those aryl groups having heteroatoms in the ring structure may also be referred to as "aryl heterocycles," "heteroaromatics" or "heteroaryls". The aromatic ring may be substituted at one or more ring positions with a substituent such as described elsewhere herein, for example, halogen, azide, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, polycyclyl, hydroxyl, alkoxy, amino, nitro, mercapto, imino, amide, phosphate, phosphonate, phosphinate, carbonyl, carboxyl, silyl, ether, alkylthio, sulfonyl, sulfonamide, ketone, aldehyde, ester, heterocyclyl, aromatic or heteroaromatic moiety, - - -CF₃- -CN, etc. The term "aryl" also includes polycyclic ring systems having two or more cyclic rings in which two or more carbons are common to two adjoining rings (the rings are "fused rings"), wherein at least one of the rings is aromatic, e.g., the additional rings can be cycloalkyls, cycloalkenyls, cycloalkynyls, aryls and/or heterocyclyls.

The term "carbocyclic" as used herein refers to an aromatic or non-aromatic ring in which each atom of the ring is carbon.

The term "carbonyl" is art-recognized and includes moieties such as may be represented by the general formula:

wherein X is a bond or represents oxygen or sulfur, and R₁₁Represents, for example but not limited to, hydrogen, alkyl, alkenyl, - - (CH)₂)_m--R₈Or a pharmaceutically acceptable salt, R'₁₁Represents hydrogen, alkyl, alkenyl or- - (CH)₂)_m--R₈Wherein m and R₈As described elsewhere herein, but not limited thereto. If X is oxygen and R₁₁Or R'₁₁Instead of hydrogen, the formula represents an "ester". If X is oxygen and R₁₁As defined above, then this moiety is referred to herein as carboxy, and especially when R is₁₁When hydrogen, the formula represents a "carboxylic acid". If X is oxygen and R'₁₁And is hydrogen, the formula represents a "formate". In general, if the oxygen atom of the above formula is replaced by sulfur, the formula represents a "thiocarbonyl". If X is sulfur and R₁₁Or R'₁₁If not hydrogen, the formula represents a "thioester". If X is sulfur and R₁₁And is hydrogen, then the formula represents "thiocarboxylic acid". If X is sulfur and R'₁₁And is hydrogen, the formula represents "thioformate". On the other hand, if X is a bond, and R₁₁Instead of hydrogen, the above formula represents a "ketone" group. If X is a bond, and R₁₁And is hydrogen, the formula represents an "aldehyde" group.

The term "electron withdrawing group" refers to a chemical group that draws electron density from the atom or group of atoms to which the electron withdrawing group is attached. Attraction of electron density includes attraction by induction and by delocalization/resonance effects. Examples of electron withdrawing groups attached to the aromatic ring include perhaloalkyl (such as trifluoromethyl), halogen, azide, carbonyl-containing groups (such as acyl), cyano, and imine-containing groups.

The term "ester" as used herein refers to a group-c (O) O-substituent, wherein the substituent table is exemplified by hydrocarbyl groups or other substituents described elsewhere herein.

The terms "halo" and "halogen" as used herein mean halogen and include chloro, fluoro, bromo, and iodo.

The term "heteroaralkyl" as used herein refers to an alkyl group substituted with a heteroaryl group.

The term "heterocycle" or "heterocyclic group" refers to a 3 to 10 membered ring structure, more preferably a 3 to 7 membered ring, which ring structure contains one to four heteroatoms. The heterocyclic ring may be polycyclic. Heterocyclic groups include, for example and without limitation: thiophene, thianthreneFuran, pyran, isobenzofuran, chromene, xanthene, phenoxathiin, pyrrole, imidazole, pyrazole, isothiazole, isoxazole, pyridine, pyrazine, pyrimidine, pyridazine, indolizine, isoindole, indole, indazole, purine, quinolizine, isoquinoline, quinoline, phthalazine, naphthyridine, quinoxaline, quinazoline, cinnoline, pteridine, carbazole, carboline, phenanthridine, acridine, pyrimidine, phenanthroline, phenazine, phenothiazine, furazan, phenoxazine, pyrrolidine, oxolane, thiacyclopentane, oxazole, piperidine, piperazine, morpholine, lactone, lactam (such as azetidinone and pyrrolidone), sultam, sultone, and the like. The heterocyclic ring may be substituted at one or more ring positions with substituents such as, but not limited to, halogen, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxy, amino, nitro, mercapto, imino, amido, phosphate, phosphonate, phosphinate, carbonyl, carboxyl, silyl, ether, alkylthio, sulfonyl, ketone, aldehyde, ester, heterocyclyl, aromatic or heteroaromatic moieties, - - -CF₃-CN, etc., and as described elsewhere herein.

The term "heteroaryl" includes substituted or unsubstituted aromatic monocyclic structures, preferably 5 to 7-membered rings, more preferably 5 to 6-membered rings, the ring structure of which comprises at least one heteroatom, preferably one to four heteroatoms, more preferably one to two heteroatoms. The term "heteroaryl" also includes up to 20-membered polycyclic ring systems having two or more rings in which two or more carbons are common to two adjoining rings, wherein at least one of the rings is heteroaromatic, e.g., the additional rings can be cycloalkyls, cycloalkenyls, cycloalkynyls, aryls, heteroaryls, and/or heterocyclyls. Heteroaryl groups include, for example, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, pyrazole, pyridine, pyrazine, pyridazine, and pyrimidine, and the like.

The term "heteroatom" as used herein means an atom of any element other than carbon or hydrogen. Preferred heteroatoms are nitrogen, oxygen and sulfur.

Thus, the terms "heterocyclyl", "heterocycle" and "heterocyclic" refer to a substituted or unsubstituted aromatic or non-aromatic ring structure (which may be a cyclic, bicyclic or fused ring system), preferably a 3 to 10 membered ring, more preferably a 3 to 7 membered ring, which ring structure contains at least one heteroatom, preferably one to four heteroatoms, more preferably one or two heteroatoms. The terms "heterocyclyl" and "heterocyclic" also include polycyclic ring systems having two or more cyclic rings in which two or more carbons are common to two adjoining rings wherein at least one of the rings is heterocyclic, e.g., the additional rings can be cycloalkyls, cycloalkenyls, cycloalkynyls, aryls, heteroaryls, and/or heterocyclyls. Heterocyclic groups include, for example, piperidine, piperazine, pyrrolidine, morpholine, lactones, lactams, and the like.

The term "5-to 20-membered heterocyclic group" or "5-to 14-membered heterocyclic group" as used throughout the present specification refers to an aromatic or non-aromatic cyclic group having 5 to 20 ring-forming atoms, preferably 5 to 14 ring-forming atoms, and containing at least one heteroatom (such as nitrogen, sulfur or oxygen) among the ring-forming atoms, which in the former case is a "5-to 20-membered, preferably 5-to 14-membered aromatic heterocyclic group" (also referred to as "heteroaryl" or "heteroaromatic") and in the latter case is a "5-to 20-membered, preferably a" 5-to 14-membered non-aromatic heterocyclic group ".

Heterocyclic groups that may be mentioned include nitrogen-containing aromatic heterocycles such as pyrrole, pyridine, pyridone, pyridazine, pyrimidine, pyrazine, pyrazole, imidazole, triazole, tetrazole, indole, isoindole, indolizine, purine, indazole, quinoline, isoquinoline, quinolizine, phthalazine, naphthyridine, quinoxaline, quinazoline, cinnoline, pteridine, imidazopyridine, imidazotriazine, pyrazinopyridazine, acridine, phenanthridine, carbazole, carbazoline, perimidine, phenanthroline, fused hexabenzene (phenacene), oxadiazole, benzimidazole, pyrrolopyridine, pyrrolopyrimidine and pyridopyrimidine; sulfur-containing aromatic heterocycles such as thiophene and benzothiophene; oxygen-containing aromatic heterocycles such as furan, pyran, cyclopentapyran, benzofuran and isobenzofuran; and aromatic heterocycles comprising 2 or more heteroatoms selected from nitrogen, sulfur and oxygen, such as thiazole, thiadiazole, isothiazole, benzoxazole, benzothiazole, benzothiadiazole, phenothiazine, isoxazole, furazan, phenoxazine, pyrazolooxazole, imidazothiazole, thienofuran, furopyrrole, pyridooxazine, furopyridine, furopyrimidine, thienopyrimidine and oxazole.

As examples of the "5 to 14-membered aromatic heterocyclic group", pyridine, triazine, pyridone, pyrimidine, imidazole, indole, quinoline, isoquinoline, quinolizine, phthalazine, naphthyridine, quinazoline, cinnoline, acridine, fused hexabenzene, thiophene, benzothiophene, furan, pyran, benzofuran, thiazole, benzothiazole, phenothiazine, pyrrolopyrimidine, furopyridine and thienopyrimidine may be preferably mentioned, and pyridine, thiophene, benzothiophene, thiazole, benzothiazole, quinoline, quinazoline, cinnoline, pyrrolopyrimidine, pyrimidine, furopyridine and thienopyrimidine may be more preferably mentioned. The term "heterocyclic group" shall generally mean a 3 to 20 membered heterocyclic group, preferably a 3 to 14 membered heterocyclic group, and all subsets of heterocyclic groups (including non-heteroaromatic or heteroaromatic) falling within the definition of heterocyclic groups are 3 to 20 membered heterocyclic groups, preferably 3 to 14 membered heterocyclic groups.

The term "8 to 20-membered heterocyclic group" or "8 to 14-membered heterocyclic group" means an aromatic or non-aromatic fused bicyclic or tricyclic group having 8 to 20, preferably 8 to 14, atoms forming a ring (two or three rings) and containing at least one heteroatom such as nitrogen, sulfur or oxygen among the atoms forming the ring, which is an "8 to 20-membered", preferably an "8 to 14-membered aromatic heterocyclic group" (also referred to as "heteroaryl" or "heteroaromatic") in the former case and an "8 to 20-membered", preferably an "8 to 14-membered non-aromatic heterocyclic group" in the latter case. The "8 to 20-membered heterocyclic group" and "8 to 14-membered heterocyclic group" are represented by the following structures: fused bicyclic, tricyclic, and tetracyclic structures containing nitrogen atoms, such as indole, isoindole, indolizine, purine, indazole, quinoline, isoquinoline, quinolizine, phthalazine, naphthyridine, quinoxaline, quinazoline, cinnoline, pteridine, imidazopyridine, imidazotriazine, pyrazinopyridazine, acridine, phenanthridine, carbazole, carbazoline, perimidine, phenanthroline, fused hexabenzene, benzimidazole, pyrrolopyridine, pyrrolopyrimidine, and pyridopyrimidine; sulfur-containing aromatic heterocycles such as thiophene and benzothiophene; oxygen-containing aromatic heterocycles such as cyclopentapyran, benzofuran and isobenzofuran; and aromatic heterocycles containing 2 or more heteroatoms selected from nitrogen, sulfur and oxygen, such as benzoxazole, benzothiazole, benzothiadiazole, phenothiazine, benzofurazan, phenoxazine, pyrazolooxazole, imidazothiazole, thienofuran, furopyrrole, pyridooxazine, furopyridine, furopyrimidine and thienopyrimidine, and the like.

The term "5-to 14-membered non-aromatic heterocyclic group" as used throughout this specification refers to a non-aromatic cyclic group having 5 to 14 ring-forming atoms and containing at least one heteroatom (such as nitrogen, sulfur or oxygen) among the ring-forming atoms. As specific examples, there may be mentioned non-aromatic heterocycles such as pyrrolidinyl, pyrrolinyl, piperidinyl, piperazinyl, N-methylpiperazinyl, imidazolinyl, pyrazolidinyl, imidazolidinyl, morpholinyl, tetrahydropyranyl, azetidinyl, oxetanyl, oxathiolanyl (oxathiolanyl), pyridone, 2-pyrrolidone, ethyleneurea, 1, 3-dioxolane, 1, 3-dioxane, 1, 4-dioxane, phthalimide and succinimide. As examples of the "5 to 14-membered non-aromatic heterocyclic group", there may be preferably mentioned pyrrolidinyl, piperidinyl and morpholinyl, more preferably pyrrolidinyl, piperidinyl, morpholinyl and pyrrole.

The term "8-to 14-membered non-aromatic heterocyclic group" as used throughout this specification refers to a non-aromatic fused ring system (typically having two or three rings) having 8 to 14 atoms forming a ring (bicyclic or tricyclic) and containing at least one heteroatom such as nitrogen, sulfur or oxygen among the atoms forming the ring.

The term "5-to 14-membered heterocyclic group" as used throughout this specification refers to an aromatic or non-aromatic cyclic group having 5 to 14 ring-forming atoms and containing at least one heteroatom (such as nitrogen, sulfur or oxygen) among the ring-forming atoms, which is a "5-to 14-membered aromatic heterocyclic group" in the former case and a "5-to 14-membered non-aromatic heterocyclic group" in the latter case. Specific examples of the "5-to 14-membered heterocyclic group" thus include specific examples of the "5-to 14-membered aromatic heterocyclic group" and specific examples of the "5-to 14-membered non-aromatic heterocyclic group".

As the "5 to 14-membered heterocyclic group", there may be preferably mentioned pyrrolidinyl, piperidinyl, morpholinyl, pyrrole, pyridine, pyridone, pyrimidine, imidazole, indole, quinoline, isoquinoline, quinolizine, phthalazine, naphthyridine, quinazoline, cinnoline, acridine, fused hexabenzene, thiophene, benzothiophene, furan, pyran, benzofuran, thiazole, benzothiazole, phenothiazine and carbostyryl (carbostyryl), more preferably pyrrolidinyl, piperidinyl, morpholinyl, pyrrole, pyridine, thiophene, benzothiophene, thiazole, benzothiazole, quinoline, quinazoline, cinnoline and carbostyryl, even more preferably thiazole, quinoline, quinazoline, cinnoline and carbostyryl, and the like.

The term "6-to 14-membered aromatic heterocyclic group" as used throughout the present specification means those substituents defined by a "5-to 14-membered aromatic heterocyclic group" having 6 to 14 ring-forming atoms. As specific examples, pyridine, pyridone, pyrimidine, indole, quinoline, isoquinoline, quinolizine, phthalazine, naphthyridine, quinazoline, cinnoline, acridine, benzothiophene, benzofuran, thiazole, benzothiazole and phenothiazine may be mentioned. "8-to 14-membered aromatic heterocyclic group" refers to those substituents or radicals having 8 to 14 atoms which form a fused bicyclic or tricyclic ring system. Specific examples include indole, quinoline, isoquinoline, quinolizine, phthalazine, naphthyridine, quinazoline, cinnoline, acridine, benzothiophene, benzofuran, benzothiazole, pyrrolopyrimidine, pyrrolopyrazine, furopyrimidine, phenothiazine and the like.

The term "6-to 14-membered heterocyclic group" as used throughout the present specification refers to those substituents defined by a "5-to 14-membered heterocyclic group" having 6 to 14 ring-forming atoms. As specific examples, mention may be made of piperidyl, piperazinyl, N-methylpiperazinyl, morpholinyl, tetrahydropyranyl, 1, 4-dioxane and phthalimide.

The term "3-to 7-membered heterocyclic group" as used throughout this specification refers to those heterocyclic substituents having from 3 to 7 ring-forming atoms, preferably from 5 to 6 ring-forming atoms.

The term "8-to 14-membered heterocyclic group" as used throughout this specification refers to those substituents defining an 8-to 14-membered heterocyclic group having 8 to 14 atoms forming a fused ring system.

The term "heterocycloalkyl" as used herein refers to an alkyl group substituted with a heterocyclic group.

The term "hydrocarbyl" as used herein refers to an optionally substituted group bonded through carbon atoms and typically having at least one carbon-hydrogen bond and a predominantly carbon chain, but which may optionally contain heteroatoms. Hydrocarbyl groups include, but are not limited to, aryl, heteroaryl, carbocycle, heterocycle, alkyl, alkenyl, alkynyl, and combinations thereof.

The term "lower" when used in connection with a chemical moiety such as acyl, acyloxy, alkyl, alkenyl, alkynyl or alkoxy is intended to encompass groups in which ten or fewer atoms, preferably six or fewer atoms, are present in the substituent. "lower alkyl" for example means an alkyl group containing ten or fewer carbon atoms, preferably six or fewer carbon atoms. In certain embodiments, acyl, acyloxy, alkyl, alkenyl, alkynyl or alkoxy substituents, as defined herein, are lower acyl, lower acyloxy, lower alkyl, lower alkenyl, lower alkynyl or lower alkoxy, respectively, whether they occur alone or in combination with other substituents, such as in the hydroxyalkyl and aralkyl lists (in which case, for example, when counting carbon atoms in an alkyl substituent, the atoms within the acyl group are not counted).

As used herein, the term "nitro" means- -NO₂(ii) a The term "halogen" denotes- -F, - -Cl, - -Br or- -I; the term "mercapto" means- -SH; the term "hydroxy" means- -OH; and the term "sulfonyl" means- -SO₂-。

The term "polycyclyl" or "polycyclic group" refers to two or more rings (e.g., cycloalkyls, cycloalkenyls, cycloalkynyls, aryls and/or heterocyclyls) in which two or more atoms are common to two adjoining rings (e.g., the rings are "fused rings"). Rings joined by non-adjacent atoms are referred to as "bridged" rings. Each of the polycyclic rings may be substituted with, but is not limited to, substituents such as those described above, e.g., halogen, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxy, amino, nitro, mercapto, imino, amido, phosphate, phosphonate, phosphinate, carbonyl, carboxy, silyl, ether, alkylthio, sulfonyl, ketone, aldehyde, ester, heterocyclyl, aromatic or heteroaromatic moieties, - - -CF3, - - -CN, and the like.

The phrase "protecting group" as used herein means a temporary substituent that avoids undesirable chemical transformation of a potentially reactive functional group. Examples of such protecting groups include esters of carboxylic acids, silyl ethers of alcohols, acetals and ketals of aldehydes and ketones. An overview of the field of protecting group chemistry has been made (Greene, T.W.; Wuts, P.G.M.Protective group in Organic Synthesis,2^nded.;Wiley:New York,1991)。

The term "substituted" refers to moieties having substituents replacing a hydrogen on one or more carbons of the backbone. It is to be understood that "substituted" or "substituted. Such substitution is in accordance with permitted valences of the atoms and substituents being substituted, and the substitution results in a stable compound, e.g., one that does not spontaneously undergo transformation, such as by rearrangement, cyclization, elimination, or the like.

As used herein, the term "substituted" is contemplated to include all permissible substituents of organic compounds. In a broad aspect, permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic, nonaromatic and inorganic substituents of organic compounds. For suitable organic compounds, the permissible substituents can be one or more and can be the same or different. For the purposes of the present invention, a heteroatom such as nitrogen may have a hydrogen substituent and/or any permissible substituents of organic compounds described herein which satisfy the valences of the heteroatom. Substituents may include any substituent (group) as described elsewhere herein, for example, halogen, hydroxyl, carbonyl (such as carboxyl, alkoxycarbonyl, formyl, or acyl), ether, thioether, thiocarbonyl (such as thioester, thioacetate, or thioformate), alkoxy, phosphoryl, phosphate, phosphonate, phosphinate, amino, amide, amidine, imine, cyano, nitro, azido, sulfhydryl, alkylthio, sulfate, sulfonate, sulfamoyl, sulfonamide, sulfonyl, heterocyclyl, aralkyl, or an aromatic or heteroaromatic moiety. One skilled in the art will appreciate that a moiety substituted on a moiety or chemical group may itself be substituted.

It is to be understood that "substituted" or "substituted. Such substitution is in accordance with permitted valences of the atoms and substituents being substituted, and the substitution results in a stable compound, e.g., one that does not spontaneously undergo transformation, such as by rearrangement, cyclization, elimination, or the like. It will be recognized that the term "unsubstituted" simply refers to a hydrogen substituent or to no substituent in the context of the use of the term.

Preferred substituents for use in the present invention include, in this context, for example, hydroxy, carboxy, cyano (C.ident.N), nitro (NO ≡ N)₂) Halogen (preferably 1,2 or 3 halogens, especially on alkyl, especially methyl, such as trifluoromethyl), thiol, alkyl (preferably C)₁-C₆More preferably C₁-C₃) Alkoxy (preferably C)₁-C₆Alkyl or aryl radicals, including phenyl), ethers (preferably C)₁-C₆Alkyl or aryl), esters (preferably C)₁-C₆Alkyl or aryl) includes alkylene esters (so as to be linked to the alkylene group, not preferably by C₁-C₆Alkyl or aryl substituted ester function), thioethers (preferably C)₁-C₆Alkyl or aryl) (preferably C)₁-C₆Alkyl or aryl), thioesters (preferably C)₁-C₆Alkyl orAryl), halogen (F, Cl, Br, I), nitro or amine (including five-or six-membered cyclic alkylene amines, including C₁-C₆Alkylamines or C₁-C₆Dialkylamine), alkanol (preferably C)₁-C₆Alkyl or aryl) or alkanoic acids (preferably C)₁-C₆Alkyl or aryl). More preferably, the term "substituted" in the context of its use shall refer to alkyl, alkoxy, halogen, hydroxy, carboxylic acid, nitro and amine (including mono-or dialkyl substituted amines). Any substitutable position in a compound according to the invention may be substituted in the present invention, but preferably no more than 5, more preferably no more than 3 substituents are present on a single ring or ring system. Preferably, the term "unsubstituted" shall mean substituted by one or more H atoms.

The term "sulfamoyl" is art-recognized and includes moieties represented by the general formula:

wherein R is₉And R₁₀Are substituents as described above.

The term "sulfate" is art recognized and includes moieties represented by the general formula:

wherein R is₄₁Is an electron pair, hydrogen, alkyl, cycloalkyl or aryl.

The term "sulfonamide" is art recognized and includes moieties represented by the general formula:

wherein R is₉And R'₁₁As described above.

The term "sulfonate" is art recognized and includes moieties represented by the general formula:

wherein R is₄₁Is an electron pair, hydrogen, alkyl, cycloalkyl or aryl.

The term "sulfoxide" or "sulfinyl" is art-recognized and includes moieties represented by the following general formula:

wherein R is₄₄Selected from hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aralkyl or aryl, which groups may be optionally substituted.

The term "thioester" is art recognized and used to describe the group-C (O) SR⁹or-SC (O) R⁹Wherein R is⁹Represents an optionally substituted hydrocarbyl group as described elsewhere herein.

As used herein, the definition of each expression of alkyl, m, n, etc., when it occurs more than once in any structure, is intended to reflect the independence of the definition of the same expression in the structure.

By way of example, certain preferred aromatic and aliphatic rings and derivatives thereof and substituents that may be used as pharmacophores or substituents in the compounds according to the invention include, but are not limited to: phenyl, benzyl, pyridine, cyclohexadiene, dihydropyridine, tetrahydropyridine, piperidine, pyrazine, tetrahydropyrazine, dihydropyrazine, piperazine, pyrimidine, dihydropyrimidine, tetrahydropyrimidine, hexahydropyrimidine, pyrimidinone, triazine, dihydrotriazine, tetrahydrotriazine, triazinane (triazazine), tetrazine, dihydrotetrazine, tetrahydrotetrazine, tetrazine (tetrazinane), pyrrole, dihydropyrrole, pyrrolidine, imidazolidine, dihydroimidazolidine, imidazole, dihydroimidazole, azetidine, triazole, dihydrotriazole, triazolidine, tetrazole, dihydrotetrazole, tetrazolidine, diazepine, diazepane, tetrahydrodiazepine, dihydrodiazepine, diazepine, oxazole, dihydrooxazole, oxazolidine, isoxazole, dihydroisoxazole, isoxazole, thiazole, dihydrothiazole, thiazolidine, isothiazole, dihydroisothiazole, isothiazole, thiazolidine, oxadiazole, oxadiazolidine, dihydropyrazine, pyrazine, triazine, pyrimidine, triazine, dihydropyrrole, thiadiazoles, dihydrothiadiazoles, thiadiazolidines, oxazinanes, dihydrooxazines (including morpholines), thiazinanes, dihydrothiazinanes, dihydrothiazines, thiazines (including thiomorpholines), thiazines, furans, dihydrofurans, tetrahydrofuran, thiophenes, pyridazine-3, 6-diones, tetrahydrothiophenes, dihydrothiophenes, tetrahydrothiophenes, dithiolanes, dithiolenes, dithiametallocenes, dioxolanes, dioxoles, oxathiolanes, pyridones, dioxanes, dioxanone diones, dihydrodioxins, dioxins, pyrans, 3, 4-dihydro 2H-pyrans, pyrones, 2H-pyran-2, 3(4H) -diones, oxathianes, dihydrooxathiadienes, oxathiahexadienes, oxathianes, thiazalines (including morpholines), thiathiazines, dihydrothiazines, dihydrothiathiazines, tetrahydrothianes, oxetane, thietane, thiazepine, cyclohexadienone, lactam, lactone, piperazinone, pyrroledione, cyclopentenone, oxazete, oxazinanone (oxazinone), dioxolane, 3, 4-dihydro-2H-thiopyran-1, 1-dioxide, dioxolane, oxazolidinone, oxazolone, thiocyclopentane-1-oxide, thiomorpholine-1-oxide, tetrahydrothiopyran, thiocyclopentane-1, 1-dioxide, dioxazine (dioxazinone), pyrazolone, 1, 3-thiazine (1,3-thiazete), thiazine-1, 1-dioxide, 6, 7-dihydro-5H-1, 4-dioxep, 1, 2-dihydropyridazin-3 (4H) -one, oxazete, thiazelate, thiazepine-1, 1-dioxide, 6, 7-dihydro-5H-1, 4-dioxep, 1, 2-dihydropyridazin-3 (4H) -one, Pyridine-2, 6(1H,3H) -dione and sugars (glucose, mannose, galactose, fucose, fructose, ribose).

Bicyclic and fused rings include, for example, naphthyl, quinone, quinolinone, dihydroquinoline, tetrahydroquinoline, naphthyridine, quinazoline, dihydroquinazoline, tetrahydroquinazoline, quinoxaline, dihydroquinazoline, tetrahydroquinazoline, pyrazine, quinazoline-2, 4(1H,3H) -dione, isoindoline-1, 3-dione, octahydropyrrolopyridine, indoline, isoindoline, hexahydroindolone, tetrahydropyrrolooxazolone, hexahydro-2H-pyrrolo [3,4-d ] isoxazole, tetrahydro-1, 6-naphthyridine, 2,3,4,5,6, 7-hexahydro-1H-pyrrolo [3,4-c ] pyridine, 1H-benzo [ d ] imidazole, octahydropyrrolo [3,4-c ] pyrrole, 3-azabicyclo [3.1.0] hexane, 7-azabicyclo [2.2.1] hept-2-ene, diazabicycloheptane, benzoxazole, indole, 1, 4-diazabicyclo [3.3.1] nonane, azabicyclooctane, naphthalene-1, 4-dione, indene, indane, 2,3,3a,7 a-tetrahydro-1H-isoindole, 2,3,3a,4,7,7 a-hexahydro-1H-isoindole, 1, 3-dihydroisobenzofuran, 1-methyl-3 a,4,5,6,7,7 a-hexahydro-1H-indole, 3-azabicyclo [4.2.0] octane, 5, 6-dihydrobenzo [ b ] thiophene, 5, 6-dihydro-4H-thieno [2,3-b ] thiopyran, 3, 4-dihydropyrazine-2 (1H) -one, 2, 3-dihydropyrazine-1H-one, 1,3, 4-dihydropyrazine-2 (1H) -one, 2, 3-dihydroindole, 2-1H-indole, 2,4, 2H-benzo [ b ] [1,4] thiazine, naphthyridin-4 (1H) -one, octahydropyrrolo [1,2-a ] pyrazine, imidazopyridazine, tetrahydroimidazopyridazine, tetrahydropyridazine, buprofezin, 5-thia-1-azabicyclo [4.2.0] oct-2-en-8-one, 4-thia-1-azabicyclo [3.2.0] hept-7-one, 1,6,7, 8-tetrahydroimidazo [4,5-d ] [1,3] diaza, 8H-thiazolo [4,3-c ] [1,4] oxazin-4-ium, 8H-thiazolo [4,3-c ] [1,4] thiazin-4-ium, pteridine, thiazolo [3,4-a ] pyrazin-4-ium, 7- (methylimino) -7H-pyrrolo [1,2-c ] thiazol-4-ium, thiazolopyrazine, 3, 7-dioxabicyclo [4.1.0] hept-4-ene, 6, 7-dihydro-5H-pyrrolo [1,2-a ] imidazole, 3 a-dihydrofuro [3,2-b ] furan-2 (6aH) -one, tetrahydro-3 aH- [1,3] dioxido [4,5-c ] pyrrole, 7-ethylene-7H-pyrrolo [1,2-c ] thiazol-4-ium, hexahydro-1H-pyrrolo [2,1-c ] [1,4] oxazine, 6,7,8,8 a-tetrahydro-1H-pyrrolo [2,1-c ] [1,4] oxazine, 2-azabicyclo [2.2.2] oct-2-ene, 6 a-dihydrothieno [3,2-b ] furan-5 (3aH) -one, 4, 5-dihydropyridin-3 (2H) -one, 4,7 a-dihydro-3 aH- [1,3] dioxido [4,5-c ] pyran, 6, 7-dihydro-1H-furo [3,4-c ] pyran-1, 3(4H) -dione, 3a,4,7 a-tetrahydro-2H-furo [2,3-b ] pyran, 2,4a,7,7 a-tetrahydro-1H-cyclopenta [ c ] pyridine, 4H-pyrano [3,2-b ] pyridine-4, 8(5H) -dione, 1,2,3,3a,4,7 a-hexahydropyrano [4,3-b ] pyrrole, 2,3,8,8 a-tetrahydroindazin-7 (1H) -one, octahydro-1H-pyrido [1,2-a ] pyrazin-1-one, 2,6,7,8,9,9 a-hexahydro-1H-pyrido [1,2-a ] pyrazin-1-one, 6,7,8,8 a-tetrahydropyrrolo [1,2-a ] pyrazin-1 (2H) -one, hexahydropyrrolo [1,2-a ] pyrazin-1 (2H) -one, bicyclo [2.2.1] hepta-2, 5-diene.

A spiro moiety: 1, 5-dioxaspiro [5.5] undecane, 1, 4-dioxaspiro [4.5] decane, 1, 4-diazabicyclo [3.2.1] octane, 5-azaspiro [2.5] octane, 5-azaspiro [2.4] heptane, 3, 9-diaza-6-azocationic spiro [5.5] undecane, 3, 4-dihydrospiro [ benzo [ b ] [1,4] oxazin-2, 1' -cyclohexane ], 7-oxa-4-azaspiro [2.5] oct-5-ene.

A pharmaceutical composition comprising a combination of an effective amount of at least one STEP modulating compound according to the invention and one or more compounds described elsewhere herein, all in effective amounts, in association with a pharmaceutically effective amount of a carrier, additive or excipient represents another aspect of the invention.

The compositions for use in the methods of treatment of the present invention and the pharmaceutical compositions of the present invention may be formulated in a conventional manner using one or more pharmaceutically acceptable carriers and may also be administered in a controlled release formulation. Pharmaceutically acceptable carriers that may be used in these pharmaceutical compositions include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins (such as human serum protein), buffer substances (such as phosphates, glycine, sorbic acid, potassium sorbate), partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes (such as prolamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts), colloidal silica, magnesium trisilicate, polyvinylpyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene-polyoxypropylene block polymers, polyethylene glycol and wool fat.

The compositions for use in the treatment methods of the invention and the pharmaceutical compositions of the invention may be administered orally, parenterally, by inhalation spray, topically, rectally, nasally, buccally, vaginally or by implantation in reservoirs. The term "parenteral" as used herein includes subcutaneous, intravenous, intramuscular, intraarticular, intrasynovial, intrasternal, intrathecal, intrahepatic, intralesional and intracranial injection or infusion techniques. Preferably, the composition is administered orally, intraperitoneally, or intravenously.

The sterile injectable dosage forms of the compositions for use in the methods of treatment of the present invention may be aqueous or oily suspensions. These suspensions may be formulated according to the techniques known in the art using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally-acceptable diluent or solvent, for example as a solution in 1, 3-butanediol. Acceptable vehicles and solvents that may be employed include water, ringer's solution 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 mono-or diglycerides. Fatty acids (such as oleic acid) and its glyceride derivatives may be used in the preparation of injectables, as may pharmaceutically-acceptable natural oils (such as olive oil or castor oil), especially in their polyoxyethylated forms. These oily solutions or suspensions may also contain a long chain alcohol diluent or dispersant, such as a swiss pharmacopoeia-grade (ph.helv) alcohol or similar alcohol.

The pharmaceutical compositions of the present invention may be administered orally in any orally acceptable dosage form including, but not limited to, capsules, tablets, aqueous suspensions or solutions. For tablets for oral administration, commonly used carriers include lactose and corn starch. Lubricating agents, such as magnesium stearate, are also typically added. For oral administration in a capsule dosage form, useful diluents include lactose and dried corn starch. When aqueous suspensions are desired for oral use, the active ingredient is combined with emulsifying and suspending agents. If desired, certain sweetening, flavoring or coloring agents may also be added.

Alternatively, the pharmaceutical compositions of the present invention may be administered in the form of suppositories for rectal administration. These suppositories can be prepared by: the medicament is mixed with a suitable non-irritating excipient which is solid at room temperature but liquid at the rectal temperature and will therefore melt in the rectum to release the drug. Such materials include cocoa butter, beeswax and polyethylene glycols.

The pharmaceutical compositions of the present invention may also be administered topically, particularly to treat skin cancer, psoriasis, or other diseases that occur in or on the skin. Suitable topical formulations can be readily prepared for each of these areas or organs. Topical application to the lower intestinal tract may be effected as a rectal suppository (see above) or in a suitable enema. Topically acceptable transdermal patches may also be used.

For topical application, the pharmaceutical compositions may be formulated in a suitable ointment containing the active ingredient suspended or dissolved in one or more carriers. Carriers for topical application of the compounds of this invention include, but are not limited to, mineral oil, liquid petrolatum, white petrolatum, propylene glycol, polyoxyethylene, polyoxypropylene compound, emulsifying wax and water.

Alternatively, the pharmaceutical compositions may be formulated in a suitable emulsion or cream containing the active ingredient suspended or dissolved in one or more pharmaceutically acceptable carriers. Suitable carriers include, but are not limited to, mineral oil, sorbitan monostearate, polysorbate 60, cetyl esters wax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol and water.

For ophthalmic use, the pharmaceutical compositions may be formulated as micronized suspensions in isotonic, pH adjusted sterile saline, or preferably as solutions in isotonic, pH adjusted sterile saline, with or without preservatives such as benzalkonium chloride. Alternatively, for ophthalmic use, the pharmaceutical composition may be formulated in an ointment (such as petrolatum).

The pharmaceutical compositions of the present invention may also be administered by nasal spray or inhalation. Such compositions may be prepared according to techniques well known in the art of pharmaceutical formulation and may be prepared as solutions in physiological saline using benzyl alcohol or other suitable preservatives, absorption promoters to enhance bioavailability, fluorocarbons, and/or other conventional solubilizing or dispersing agents.

The amount of compound in the pharmaceutical composition of the invention that can be combined with a carrier material to produce a single dosage form will vary depending upon the host and the disease to be treated, the particular mode of administration. Preferably, a composition should be formulated containing from about 0.05 mg to about 750 mg or more, more preferably from about 1mg to about 600 mg, even more preferably from about 10mg to about 500mg of the active ingredient alone or in combination with at least one additional non-antibody attracting compound useful for the treatment of cancer, prostate cancer or metastatic prostate cancer or secondary effects or disorders thereof.

It will also be understood that the specific dose and treatment regimen for any particular patient will depend upon a variety of factors including the activity of the specific compound employed, the age, body weight, general health, sex, diet, time of administration, rate of excretion, drug combination, the judgment of the treating physician, and the severity of the particular disease or disorder being treated.

A patient or subject (e.g., a man) having or at risk of developing a neurological disorder can be treated by: an effective amount of (-) -huperzine a and related aspects and embodiments (including pharmaceutically acceptable salts, solvates, or polymorphs thereof) according to the present invention is administered to a patient (subject) for treatment, optionally in a pharmaceutically acceptable carrier or diluent, alone or in combination with other known agents, preferably agents that can assist in the treatment of neurological disorders or ameliorate secondary effects and conditions associated with neurological disorders. The present treatment may also be administered in conjunction with other conventional therapies, such as drugs for treating cognitive and behavioral symptoms in alzheimer's patients (e.g., drug therapy for treating cognitive and behavioral symptoms in alzheimer's disease patientsAnd）。

the compounds may be administered by any suitable route, for example orally, parenterally, intravenously, intradermally, subcutaneously or topically in liquid, cream, gel or solid dosage forms or by aerosol dosage forms.

The active compound is included in a pharmaceutically acceptable carrier or diluent in an amount sufficient to deliver a therapeutically effective amount to the desired indication in a patient without causing serious toxic side effects in the patient being treated. For all conditions mentioned herein, preferred dosages of the active compound are in the range of from about 10ng/kg to 300mg/kg per day, preferably from 0.1 to 100mg/kg per day, more usually from 0.5 to about 25mg per kg of receptor per patient body weight per day. Typical topical dosages will be in the range of 0.01-3% wt/wt in a suitable carrier.

The compounds are typically administered in any suitable unit dosage form, including, but not limited to, unit dosage forms containing less than 1mg, 1mg to 3000mg, preferably 5 to 500mg, of active ingredient per unit dosage form. Oral dosages of about 25-250mg are generally suitable.

The active ingredient is preferably administered to achieve a peak plasma concentration of the active compound of about 0.00001-30mM, preferably about 0.1-30 μ M. This can be achieved, for example, by intravenous injection of a solution or formulation of the active ingredient, optionally in saline or an aqueous medium, or administration as a bolus (bolus) of the active ingredient. Oral administration is also suitable for producing effective plasma concentrations of the active agent as with topically applied compositions.

The concentration of the active compound in the pharmaceutical composition will depend on absorption, distribution, inactivation, and rate of excretion of the drug, as well as other factors known to those skilled in the art. It should be noted that dosage values will also vary with the severity of the condition to be alleviated. It is also to be understood that for any particular subject, the specific dosage regimen will be adjusted over time according to the individual needs and the professional judgment of the person administering or supervising the administration of the composition, and that the concentration ranges set forth herein are exemplary only and are not intended to limit the scope or practice of the claimed compositions. The active ingredient may be administered at one time, or may be divided into a plurality of smaller doses to be administered at different time intervals.

The oral composition will typically comprise an inert diluent or an edible carrier. They may be encapsulated in gelatin capsules or compressed into tablets. For the purpose of oral therapeutic administration, the active compound or prodrug derivative thereof may be compounded with excipients and used in the form of tablets, troches or capsules. Pharmaceutically compatible binding agents and/or adjuvant materials may be included as part of the composition.

Tablets, pills, capsules, lozenges and the like may comprise any of the following ingredients or compounds of similar nature: a binder such as microcrystalline cellulose, gum tragacanth or gelatin; excipients, such as starch or lactose; dispersing agents such as alginic acid, Primogel or corn starch; lubricants, such as magnesium stearate or Sterotes; glidants, such as colloidal silicon dioxide; sweetening agents, such as sucrose or saccharin; or a flavoring agent such as peppermint, methyl salicylate, or orange flavoring. When the dosage unit form is a capsule, it may contain, in addition to materials of the above type, a liquid carrier such as a fatty oil. In addition, the dosage unit forms may contain various other materials which modify the physical form of the dosage unit, such as sugar, shellac, or enteric coatings.

The active compound or a pharmaceutically acceptable salt thereof may be administered as a component of an elixir, suspension, syrup, wafer, chewing gum or the like. Syrups may contain, in addition to the active compounds, sucrose as a sweetening agent and certain preservatives, pigments and coloring and flavoring agents.

The active compound or a pharmaceutically acceptable salt thereof may also be mixed with other active materials that do not harm the desired effect or with other materials that supplement the desired effect, such as other anticancer, antibiotic, antifungal, anti-inflammatory or antiviral compounds.

Solutions or suspensions for parenteral, intradermal, subcutaneous or topical application may comprise the following components: sterile diluents such as water for injection, saline solution, fixed oils, polyethylene glycols, glycerin, propylene glycol or other synthetic solvents; antibacterial agents such as benzyl alcohol or methyl paraben; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents, such as ethylenediaminetetraacetic acid; buffers such as acetate, citrate or phosphate; and agents to adjust tonicity such as sodium chloride or dextrose. The parenteral formulations may be enclosed in ampoules, disposable syringes or multiple dose vials made of glass or plastic.

If administered intravenously, the preferred carrier is physiological saline or Phosphate Buffered Saline (PBS).

In one embodiment, the active compound is formulated with a carrier that will avoid rapid elimination of the compound from the body, such as a controlled release formulation, including implants and microencapsulated delivery systems. Biodegradable biocompatible polymers such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid may be used. Methods of formulating such formulations will be apparent to those skilled in the art.

Liposomal suspensions may also be pharmaceutically acceptable carriers. They can be prepared according to methods known to those skilled in the art, for example, as described in U.S. Pat. No. 4,522,811 (which is incorporated herein by reference in its entirety). For example, liposomal formulations can be prepared by: suitable lipids such as stearoyl phosphatidyl ethanolamine, stearoyl phosphatidyl choline, arachidoyl phosphatidyl choline and cholesterol are dissolved in an inorganic solvent, and the solvent is then evaporated, leaving a dry lipid film on the surface of the container. An aqueous solution of the active compound is then introduced into the vessel. The vessel was then vortexed by hand to detach the lipid material from the vessel sidewall and disperse the lipid aggregates, thereby obtaining a liposome suspension.

Exemplary methods and Compounds of the invention

Scheme 1 below illustrates one preferred synthesis within the scope of the present invention.

The descriptors a, b, c represent the steps carried out by batch aqueous workup of the product without purification of the product. Steps 3a and 3d were completed by evaporation of volatiles and direct addition of reagents to the reaction flask (no batch post-treatment). The route to synthesis 5 proceeds from 1 in 55% overall yield.

This route can start with the enantiomerically pure reagent (+) -pulegone at low cost, which can be converted into 1 by a four-step procedure, which has been published previously. Lee, h.w.; Ji, s.k.; Lee i. -y.c.; Lee j.h.j.org.chem.1996,61,254.

There are many new features associated with exemplary scenario 1. For example, the existing stereochemistry of the starting material may be used to control the relative stereochemistry in product 2.

The known art does not suggest the conversion of 2 to 3. In particular, step 2b is the first known example of β -ketonitrile cyclization.

The known art also does not suggest the Wittig reaction (step 2 c). Previous workers have obtained mixtures of olefin isomers in the product. We have optimized the substrates and reaction conditions to achieve the desired results.

The known art also does not suggest the conversion of 4 to 5. Previous workers relied on a two-step procedure to eliminate the alcohol function (step 4 a). We have found that this can be performed efficiently in one reaction flask using a bergels reagent.

Step 4b is also not proposed by the known art. The use of platinum catalysts in the hydration of nitriles to primary amides is rare. In addition, it is known in the literature that such catalysts are generally ineffective for the hydration of tertiary nitriles (such as present in 4). Thus, the direct hydration of nitriles using this catalyst constitutes an advance in the known synthesis processes.

Intermediates 3,4 and 5 can potentially be used to obtain other natural products. In particular, compounds such as 5 may be useful scaffolds for the synthesis of other biologically active compounds.

The significant improvement in yield and the significant reduction in process steps achieved by the present invention are demonstrated by comparison with known processes as outlined below.

Summary of previous syntheses:

1.Xia,Y.;Kozikowski,A.P.J.Am.Chem.Soc.1989,111,4116.

2.Qian,L.;Ji,R.Tetrahedron Lett.1989,30,2089.

3.Yamada,F.;Kozikowski,A.P.;Reddy,E.R.;Pang,Y.P.;Miller,J.H.;McKinney,M.J.Am.Chem.Soc.1991,113,4695.

4.Lucey,C.;Kelly,S.A.;Mann,J.Org.Biomol.Chem.2007,5,301.

5.Koshiba,T.;Yokoshima,S.;Fukuyama,T.Org.Lett.2009,11,5354.

further description of the process of the invention

Most methods for huperzine a synthesis rely on the introduction of a four carbon segment into the bicyclic structure (retro-synthetic cleavage of the a and b bonds in 1 as shown below).

Reverse synthetic analysis of (-) -huperzine A (1).

Scheme 2 below provides further details on the chemical techniques used in the scheme 1 process as described above, and points out the unseparated intermediates generated in each step of our process.

Referring to schemes 1 and 2, we developed a novel process in which the cleavage of two alternative bonds (see 2 in "(-) -huperzine A (1) retro-Synthesis analysis" above) results on the basis of ketone and ketone respectivelySynthons 3 and 4 for pyridones. The former is obtainable from (R) -4-methyl-cyclohex-2-en-1-one (5) and 3-bromo-2- (bromomethyl) -6-methoxypyridine (6) will be the functional equivalent of 4. The C-4 stereocenter in 5 was used in our route to control the relative and absolute stereochemistry in the target. Various convenient methods for preparing (R) -4-methylcyclohex-2-en-1-one (5) have been reported.¹⁶In a preferred embodiment, a direct four-step sequence starting from (+) -pulegone may be used.^16aDihalopyridines such as 6 have been used in different and significantly longer routes to the synthesis of (-) -huperzine a (1), 14c and in the synthesis of other huperzine (Lycopodium) alkaloids.¹⁷

Scheme 2 enantioselective synthesis of (-) -huperzine A (1). Reagents and conditions:

(1)Ph(CH₃)₂SiLi, CuI, HMPA, THF, -78 → -23 → -78 ℃, then 6, -78 → -23 ℃, 84-91%; (2a) LHMDS, p-TsCN, toluene, -78 ℃; (2b) pd (Pt-Bu)₃)₂(5 mol%) NaOt-Bu, toluene, 110 ℃; (2c) EtPPh₃Br，LHMDS，Et₂O, 24 ℃, 71-76% from 7, E: Z =5: 1; (3a) TfOH, DCM, 0 → 24 ℃ and then TBAF, H₂O₂，K₂CO₃DMF, 40 ℃, E: Z =5: 1; (3b)12, toluene, 110 ℃, E: Z =5: 1; (3c)13(2 mol%), EtOH-H₂O(2:1)，95℃，E:Z=5:1；(3d)PIFA，CH₃OH, reflux, then TMSI, CHCl₃Refluxing, then CH₃OH, reflux, 56-70% from 10.

Successful implementation of this strategy is shown in scheme 2 above. To make this route suitable for large scale synthesis, we have extensively optimized each step, which effectively simplifies many transformations (the final synthetic route requires three chromatographic purification steps). Our work began with the conjugate addition of lithium dimethylphenylsilylcuprate with (R) -4-methyl-cyclohex-2-en-1-one (5). Alkylation of the initial enolate by 3-bromo-2- (bromomethyl) -6-methoxypyridine (6) gave addition-alkylation product 7 as the single detectable diastereomer (1H NMR analysis), isolated after purification (2.0-4.5 g scale) in 84-91% yield.

The kinetics of 7 control the deprotonation and the capture of the resulting enolate by p-toluenesulfonylcyanide,¹⁸the product mixture is then subjected to immediate work-up to high purity (estimated)>95%，¹H NMR analysis) α -cyanoketone 8 was formed rapid isolation of the product was critical because α -cyanoketone 8 would undergo disproportionation to starting material (7) and α -dicyanolone (not shown) if the mixture was allowed to age.

Unpurified α -cyanoketone 8 was then subjected to palladium-catalyzed intramolecular enolate heteroarylation.¹⁹Bis (tri-tert-butylphosphino) palladium (0) prepared by the methods of Dai and Fu among the various catalyst precursors studied²⁰As the most efficient occurrence. A dramatic dependence on base was observed (table 1).

Table 1: optimization of enolate heteroarylation. ^a

^aPd (Pt-Bu) was used for all reactions₃)₂As precatalyst in toluene at 110 ℃ for 3 h.^bIsolated yield after purification by flash column chromatography.^cBy unpurified reaction mixture¹H NMR and LC/MS analysis. Dec. = decomposition.

Thus, in the presence of carbonate base (entries 1, 2), the prototype debrominated (protodebrominated) product 15 dominates. Sodium hydride (entry 3) increased the conversion of the cyclization product (9), however, a large amount of decomposition also occurred. Finally, we determined that sodium tert-butoxide (entry 5) is optimalAnd the product was obtained in substantially quantitative yield (1H NMR analysis) using this base. The next step of the sequence requires stereoselective olefination of the ketone function of 9. The olefination product 10 is obtained in high yield by lithium ylide treatment 9 (ether, 24 ℃) derived from ethyltriphenylphosphonium bromide. A clear trend was observed between E: Z selectivity and concentration (E/Z1.1, 1.8, 5 at 1.0, 0.1 and 0.01M respectively), consistent with the salt effect and indicating that the desired E-isomer is a kinetically favored product.²¹Under optimized conditions, the olefination product 10 was isolated from 7 by flash column chromatography (4.3-7.4 g scale) as a 5:1 mixture of E/Z isomers in 71-76% yield. By this method, a complete carbon skeleton of 1 is formed in four steps in a high overall yield and on a several gram scale.

Treatment of the alkylene product (10) with trifluoromethanesulfonic acid followed by oxidative desilylation affords high purity (A)¹HNMR analysis) of cyanohydrin 11. The unpurified cyanohydrin 11 is effectively dehydrated by heating in toluene with a berges reagent (12). In the presence of a platinum catalyst 13 in aqueous ethanol²²The pyrolysis (not shown) of the dehydration product is carried out in the case of (1) to obtain the amide 14. Finally, Hofmann rearrangement of [ bis (trifluoroacetoxy) iodobenzene]Complete deprotection and purification by flash column chromatography gave (-) -huperzine a alone (1, 56-70% via four runs) and its alkene isomer (not shown, 11-14%). Synthetic (-) -huperzine A (1) in all respects (1)¹H NMR, 13C NMR, IR, three TLC solvent systems, LC/MS retention time, optical rotation) were all the same as for the real sample. Batches of (-) -1 up to 1.6g have been prepared.

To date, more than 3.5g of (-) -huperzine A (1) has been prepared by the route outlined above. Our synthesis was performed in 35-45% overall yield (16 times more efficient than any other previously reported enantioselective route) and required only three chromatographic purifications. We expect that this chemistry will reliably supply synthetic (-) -huperzine a (1) and will greatly facilitate its clinical development in neuroprotective and anti-neurodegenerative applications.

One of ordinary skill in the art will recognize that the various reactants, reagents, and reactions used in the methods of the invention can be varied in many ways without affecting the efficiencies and yields as described herein.

For example, the formation of (. + -.) huperzine from amide 14 can be generally described in accordance with Loudon, G.M., Radhakrishna, A.S., Almount, M.R., Blodgett, J.K., Boutin, R.H.J.Org.Chem.,1984,49, 4272-4276; the process described in Zhang, L.; Kaufmann, G.S.; Pesti, J.A.; Yin, J.J.org.Chem.,1997,62, 6918-. Ethanol, propanol or water may replace methanol in the hofmann rearrangement of amide 14.

Dehydration of cyanohydrin 11 using berges reagent can be accomplished in a number of ways, for example by using the techniques described in k.c. nicolaou, d.y. -k.chen, x.huang, t.ling, m.bella, s.a.snyder, "Chemistry and biology of Diazonamide a: First Total Synthesis and validation of the truetstruture" j.am. chem.soc.126,12888-12896 (2004).

The conversion of the olefination product 10 to cyanohydrin 11 as described herein can be achieved in a variety of ways. For example, desilylation can be achieved by reaction with boron trifluoride-acetic acid complex or a bronsted acid such as TFA, MSA, FMSA or tetrafluoroboric acid in an inert solvent such as DCM. When boron trifluoride-acetic acid complex is used, the alkylene product 10 may be treated with hydrogen peroxide and KHCO₃And (4) oxidizing. When Bronsted acids are used, the alkylene product 10 may be hydrogen peroxide, KHCO₃And KF oxidation. Methods useful for converting silyl groups to hydroxyl groups are also described in Fleming, i. (Chemgrams-Organic Chemistry 1996,9,1-64) and Jones, G.R.et al (Tetrahedron,1996,52, 7599-7662).

The Wittig olefination reaction used to convert the cyclized product 9 to the olefination product 10 can be improved in a number of ways, for example, by using a base such as n-butyllithium, sodium bis (trimethylsilyl) amide, lithium bis (trimethylsilyl) amide, potassium bis (trimethylsilyl) amide, or lithium diisopropylamide in a solvent such as THF, diethyl ether, or 1, 4-dioxane. See chem.Rev.1989,89,863, Modem Carbonyl elongation 2004,1-17, LiebigsAnn. chem.1997, 1283.

In the cyanation of the enolate formed by deprotonation of the addition-alkylation product 7, the toluene in the cyanation reaction can be replaced by THF. See d.kahne and d.b.columm, Tetrahedron lett.,5011(1981), and Lithium Diisopropylamide (LDA) may be replaced with lithium bis (trimethylsilyl) amide (LHMDS).

Any of the methods described in the reference noted 16 can be used to prepare the starting material (R) -4-methyl-cyclohex-2-en-1-one 1.

Further details regarding the above-described methods are given in the exemplary experimental section below.

Experimental section

General experimental procedure. All reactions were carried out under positive argon pressure in a single-necked, flame-dried round-bottomed flask equipped with a rubber septum, unless otherwise indicated. Air and moisture sensitive liquids are transferred by syringe or stainless steel cannula, or treated in a nitrogen-filled dry box (working oxygen content)<1 ppm). The organic solution was concentrated by rotary evaporation at 30-33 ℃. Such as Still et al¹The silica gel obtained from Sorbent Technologies (Atlanta, GA) was used: (40-63 μm particle size) was subjected to flash column chromatography. Silica gel impregnated with a fluorescent indicator (254nm) (1.0 mm,pore size) pre-coated glass plates were subjected to analytical Thin Layer Chromatography (TLC). TLC plates were prepared by exposure to Ultraviolet (UV) light or/and immersion in aqueous potassium permanganate solution (KMnO)₄) Then briefly heated on a hot plate (120 ℃ C., 10-15 s) for observation.

A material. Commercial solvents and reagents were used as received with the following exceptions. Benzene, dichloromethane, ether and toluene were purified according to Pangborn et al.²Tetrahydrofuran was distilled from sodium/benzophenone under nitrogen atmosphere just before use. Methanol was distilled from magnesium methoxide under nitrogen atmosphere immediately before use. Hexamethylphosphoramide was distilled from calcium hydride under nitrogen and stored under nitrogen. Will be provided withThe molecular sieves were activated by heating under vacuum overnight (200 ℃,200 mtorr), stored in a gravity oven (120 ℃), and flame dried under vacuum (100 mtorr) immediately prior to use. A solution of phenyldimethylsilyl lithium in tetrahydrofuran was prepared according to the procedure of Fleming and co-workers.³(R) -4-methyl-cyclohex-2-en-1-one (5) was prepared from (+) -pulegone according to the procedure of Lee and co-workers.⁴3-bromo-2- (bromomethyl) -6-methoxypyridine (6) was prepared according to the procedure of Kelly and co-workers.⁵Bis (tri-tert-butylphosphino) palladium (0) was prepared according to the procedures of Dai and Fu.⁶Methyl N- (triethylammoniunsulfonyl) carbamate (Burgis reagent, 12) was prepared according to the procedure of Burgess and co-workers.⁷Hydrogenated (hydroxydimethylphosphine) [ Hydrobis (hydroxydimethylphosphine) was prepared according to the procedure of Ghaffar and Parkins]Platinum (II) (13).⁸Ethyltriphenylphosphonium bromide was recrystallized in water, and the resulting crystals were dried in vacuo at 50 ℃ for 24 h.

An apparatus. Proton nuclear magnetic resonance spectroscopy (unless otherwise indicated) (ii)¹H NMR) were recorded at 400 or 500MHz and 24 ℃. Chemical shifts are expressed in parts per million (ppm, coordinate) at the tetramethylsilane low field and are referenced to NMR solvent (CHCl)₃7.26). The data are presented below: chemical shifts, multiplicity (s = singlet, d = doublet, t = triplet, q = quartet, m = multiplet and/or multiple resonance, br = broad, app = apparent), integral, coupling constant (hertz) and attribution. Proton decoupled carbon nuclear magnetic resonance spectroscopy, unless otherwise indicated (¹³C NMR) are recorded at 100 or 125MHz and 24 ℃. Chemical shifts are expressed in parts per million (ppm, coordinate) at the tetramethylsilane low field and are referenced to the solvent (CDCl)₃77.0). Distortion-free enhancement of polarization-transferred spectra [ DEPT (135) ]]Both recorded at 100 or 125MHz and 24 ℃. Will be provided with¹³The C NMR and DEPT (135) data were combined and are shown below: chemical Shift, carbon type [ from DEPT (135) experiment]. Attenuated Total reflectance Fourier transform Infrared Spectroscopy (ATR-FTIR) was obtained using a Thermo Electron Corporation Nicolet6700FTIR spectrometer referenced to polystyrene standards. The data are presented below: absorption frequency (cm)^-1) Absorption intensity (s = strong, m = medium, w = weak, br = wide). High Resolution Mass Spectrometry (HRMS) data were obtained using a Waters UPLC/HRMS instrument equipped with a dual API/ESI high resolution mass spectrometry detector and a photodiode array detector. Unless otherwise indicated, samples were in reverse phase C₁₈The linear gradient of 5% acetonitrile-water containing 0.1% formic acid → 95% acetonitrile-water containing 0.1% formic acid on a column (1.7 μm particle size, 2.1 × 50 mm) over 4 minutes and then eluted at a flow rate of 600 μ L/min over 1 minute through 100% acetonitrile containing 0.1% formic acid the optical rotation was measured on a Perkin Elmer polarimeter equipped with a sodium (589nm, D) lamp the optical rotation data is expressed as the specific optical rotation ([ α ] m particle size, 2.1 × mm)]²⁰ _n) Concentration (g/mL) and solvent.

And (4) synthesizing procedures.⁹

Step 1: addition-alkylation of (R) -4-methyl-cyclohex-2-en-1-one (5) (addition-alkylation product 7):

hexamethylphosphoramide (11.4 mL, 65.4mmol, 3.60 equiv.) was added dropwise via syringe to a stirred suspension of cuprous iodide (3.46 g, 18.2mmol, 1.00 equiv.) in tetrahydrofuran (36mL) at 24 ℃. The resulting mixture was cooled to-78 ℃. A solution of lithium dimethylphenylsilyl in tetrahydrofuran (0.46M, 79.0mL, 36.3mmol, 2.00 equiv.) was added dropwise to the cold brown suspension over 30 minutes by syringe pump. After the addition was complete, the mixture was warmed to 0 ℃. The resulting solution was stirred at 0 ℃ for 1 h. The mixture was then cooled to-78 ℃. (R) -4-methyl-cyclohex-2-en-1-one (5, 2.00g, 18.2mmol, 1.00 eq.) was added dropwise via syringe over 5 minutes. After the addition was complete, the reaction mixture was warmed to-23 ℃. The warm solution was stirred at-23 ℃ for 3 h. The reaction mixture was then cooled to-78 ℃. A solution of 3-bromo-2- (bromomethyl) -6-methoxypyridine (6) in tetrahydrofuran (0.50M, 40.0mL, 20.0mmol, 1.10 equiv.) was added dropwise to the cold reaction mixture over 30 minutes via a cannula. After the addition was complete, the reaction mixture was warmed to-23 ℃. The warm solution was stirred at-23 ℃ for 1 h. The product mixture was then warmed to 24 ℃ over 30 minutes. The warm product mixture was eluted through a pad of celite (length/diameter =3/9 cm). The celite pad was washed with a saturated aqueous sodium bicarbonate solution (100mL), ethyl acetate (250mL), a saturated aqueous sodium bicarbonate solution (100mL), and ethyl acetate (250mL) in this order. The biphasic filtrate was collected and transferred to a separatory funnel. The formed layers are separated. The organic layer was washed successively with a saturated aqueous sodium bicarbonate solution (2X 200mL), distilled water (200mL) and a saturated aqueous sodium chloride solution (200 mL). The washed organic layer was dried over sodium sulfate. The dried solution was filtered and the filtrate was concentrated. The resulting residue was purified by flash column chromatography (eluting with 5% ethyl acetate-hexanes) to afford the addition-alkylation product 7 as a light yellow viscous oil (7.37g, 91%).

R_f=0.27 (5% ethyl acetate-hexanes, KMnO)₄）。[α]²⁰ _n=-40.8(c0.10,CHCl₃)。¹H NMR(500MHz,CDCl₃),7.55(d,1H,J=8.5Hz,H₁),7.45(dd,2H,J=8.0,2.0Hz,H₁₂),7.33-7.29(m,3H,H₁₂),6.42(d,1H,J=8.5Hz,H₂),3.79(s^-,3H,H₃),3.22-3.12(m,2H,H₄/H₅),2.84(dd,1H,14.5,4.5Hz,H₄),2.58-2.52(m,1H,H₉),2.23-2.17(m,1H,H₉),2.05-1.94(m,2H,H₇/H₈),1.82-1.75(m,1H,H₈),1.15(t,1H,J=6.5Hz,H₆),1.00(d,3H,6.5Hz,H₁₀),0.32(app s,6H,H₁₁)。¹³C NMR(125MHz,CDCl₃),214.8(C),162.2(C),154.7(C),142.4(CH),138.1(C),134.0(CH),129.3(CH),128.0(CH),112.2(C),110.1(CH),53.6(CH₃),47.1(CH),40.3(CH₂),37.3(CH₂),34.3(CH),31.1(CH₂),29.3(CH),23.9(CH₃),-3.0(CH₃),-3.6(CH₃)。IR(ATR-FTIR),cm^-1:2951(br),1709(s),1575(s),1459(s),1417(s),1295(m),1250(m),1111(m),1037(m),1014(m),820(s),734(m),701(m)。HRMS-CI(m/z):[M+H]⁺C₂₂H₂₉BrNO₂Si calculated 446.1146/448.1125; found 446.1147/448.1124.

Step 2 a-c: synthesis of alkylene product 10:

step 2 a: cyanation of addition-alkylation product 7(α -cyanoketone 8):

a solution of lithium hexamethyldisilazide in toluene (1.00M, 49.7mL, 49.7mmol, 3.00 equiv.) was added dropwise via syringe pump to a stirred solution of addition-alkylation product 7(7.37g, 16.6mmol, 1.00 equiv.) in toluene (170mL) at-78 deg.C over 15 minutes. After the addition was complete, the reaction mixture was warmed to 0 ℃. The warm solution was stirred at 0 ℃ for 15 minutes. The mixture was then cooled to-78 ℃. A toluene solution of p-toluenesulfonylnitrile (1.00M, 18.2mL, 18.2mmol, 1.10 equiv.) was quickly injected by a syringe: (<1 minute) was added to the cold reaction mixture, the reaction mixture was stirred at-78 ℃ for 1 minute, the cold product mixture was rapidly diluted with 100mM aqueous sodium phosphate buffer (pH7,30mL), the product mixture was allowed to warm to 24 ℃ over 30 minutes with stirring, the warmed product mixture was diluted with ethyl acetate (200mL), the diluted product mixture was transferred to a separatory funnel already containing 100mM aqueous sodium phosphate buffer (pH7,150mL), the resulting layer was separated, the aqueous layer was extracted with ethyl acetate (3 × 150mL)) Extract, combine the organic layers, dry the combined organic layers over sodium sulfate, filter the dried solution, and concentrate the filtrate to afford unpurified α -cyanoketone 8 as a pale yellow viscous oil.¹H NMR analysis (400MHz, CDCl)₃) Show that>95% conversion to cyanoketone 8 ((R) - α -cyanoketone, (S) - α -cyanoketone and β -hydroxy- α -unsaturated nitrile isomer mixture).

Alpha-cyanoketone 8 was found to be unstable for flash column chromatography purification. Therefore, no further characterization was attempted.

And step 2 b: cyclization of α -cyanoketone 8 (tricyclo 9):

to a 500mL round bottom flask attached to a teflon coated valve was added unpurified α -cyanoketone 8 (16.6 mmol, 1.00 equiv., assuming quantitative yield in the previous step.) the residue was dried by azeotropic distillation with benzene (5.0 mL.) the vessel was sealed and the sealed vessel was then transferred to a nitrogen filled dry box sodium tert-butoxide (1.75 g, 18.2mmol, 1.10 equiv.), bis (tri-tert-butylphosphine) palladium (0) (423 mg, 828 μmol, 0.05 equiv.) and toluene (170mL) were added to the flask in that order.¹H NMR analysis (400MHz, CDCl)₃) Indicating the formation of the cyclization product9 conversion rate>95 percent. The product thus obtained was used directly in the next step. An analytically pure sample of the cyclized product 9 was obtained by flash column chromatography (eluting with 5% ethyl acetate-hexanes):

R_f=0.23 (5% ethyl acetate-hexane, KMnO)₄）。¹H NMR(500MHz,CDCl₃),7.64(d,1H,J=9.0Hz,H₁,7.51(dd,2H,J=7.0,1.5Hz,H₁₁),7.39-7.29(m,3H,H₁₁),6.74(d,1H,J=8.5Hz,H₂),3.91(s,3H,H₃),3.14(dd,1H,7=18.0,4.5Hz,H₄),2.95-2.92(m,1H,H₅),2.82-2.77(m,2H,H₄/H₈),2.15(dd,1H,J=13.5,10.0Hz,H₈),1.85-1.78(m,1H,H₇),1.32(dd,1H,J=10.0,6.5Hz,H₆),0.75(d,3H,J=6.5Hz,H₉),0.40(s,3H,H₁₀),0.37(s,3H,H₁₀)。¹³C NMR(125MHz,CDCl₃),206.0(C),164.1(C),149.5(C),138.5(CH),136.9(C),134.1(CH),129.8(CH),128.3(CH),125.1(C),119.2(C),111.0(CH),53.9(CH₃),52.4(CH₂),49.9(C),44.9(CH),42.4(CH₂),38.1(CH),28.2(CH),21.8(CH₃),-3.4(CH₃),-3.8(CH₃)。IR(ATR-FTIR),cm^-1:2955(br),2268(w),1736(s),1713(w),1599(m),1576(w),1476(s),1424(m),1321(m),1264(m),1130(m),1112(m),1028(m),824(s),737(w),704(m)。HRMS-CI(m/z):[M+H]⁺C₂₃H₂₇N₂O₂Calculated Si 391.1837; found 391.1839.

And step 2 c: olefination of cyclization product 9 (alkene 10):

in a dry box filled with nitrogen, ethyltriphenylphosphonium bromide (7.38 g, 19.9mmol, 1.20 equiv.) and lithium hexamethyldisilazide (3.33 g, 19.9mmol, 1.20 equiv.) were added sequentially to a 500mL round bottom flask. The flask was sealed with a rubber septum, and the sealed flask was then taken out of the dry box. Ether (200mL) was added to the flask via syringe. The resulting orange suspension was stirred at 24 ℃ for 1 h. During this time, the solid dissolved to form a clear orange solution. In a separate flask, a solution of unpurified cyclized product 9 (16.6 mmol, 1.00 equiv, assuming quantitative yield in the previous step) in ether (1.5L) was prepared. The orange ylide solution was transferred via cannula over 10 minutes to the flask containing the cyclized product 9 at 24 ℃. The reaction mixture was stirred at 24 ℃ for 12 h. The product mixture was poured into a separatory funnel that had been filled with distilled water (500mL) and ethyl acetate (500 mL). The formed layers are separated. The aqueous layer was extracted with ethyl acetate (2X 500 mL). The organic layers were combined and the combined organic layers were dried over sodium sulfate. The dried solution was filtered and the filtrate was concentrated. The resulting residue was purified by flash column chromatography (eluting with 5% ethyl acetate-hexanes) to give the olefination product 10 as a pale yellow viscous oil (4.74 g, 71% from 7, 5:1 mixture of E/Z diastereomers).

R_f=0.20 (5% ethyl acetate-hexanes, KMnO)₄）。¹H NMR（400MHz，CDCl₃5:1 mixture of diastereomers): e-olefin (major diastereomer), 7.69(d,1H, J =8.4Hz, H₁),7.54-7.48(m,2H,H₁₁),7.39-7.34(m,3H,H₁₁),6.64(d,1H,J=8.8Hz,H₂),5.95(q,1H,J=6.8Hz,H₁₂),3.90(s,3H,H₃),3.37-3.34(m,1H,H₅),2.86(dd,1H,J=17.6,4.8Hz,H₄),2.60-2.55(m,1H,H₄),2.50(dd,1H,J=12.4,6.0Hz,H₈),1.79-1.68(m,2H,H₇/H₈),1.72(d,3H,J=6.8Hz,H₁₃),0.77(dd,1H,J=8.8,5.6Hz,H₆),0.63(d,3H,J=6.8Hz,H₉),0.37(s,3H,H₁₀),0.36(s,3H,H₁₀) (ii) a Z-olefin (minor diastereomer), 7.78(d,1H, J =8.8Hz, H₁),7.54-7.48(m,2H,H₁₁),7.39-7.34(m,3H,H₁₁),6.67(d,1H,J=8.8Hz,H₂),5.60(q,1H,J=7.6Hz,H₁₂),3.91(s,3H,H₃),2.94(dd,1H,J=17.6,4.8Hz,H₄),2.75-2.70(m,1H,H₅),2.62-2.46(m,2H,H₄/H₈,2.02(d,3H,J=8Hz,H₁₃),1.79-1.68(m,2H,H₇/H₈),0.67-0.60(m,1H,H₆),0.62(d,3H,J=6Hz,H₉),0.36(s,3H,H₁₀),0.33(s,3H,H₁₀)。¹³C NMR（100MHz，CDCl₃5:1 mixture of diastereomers): e-olefins (major diastereomer), 163.3(C),151.9(C),138.3(C),137.9(CH),134.2(C),134.0(CH),129.4(CH),128.1(CH),127.4(C),122.7(C),118.2(CH),109.5(CH),53.7 (CH)₃),50.4(CH₂),44.4(C),42.2(CH₂),34.7(CH),30.7(C),27.7(CH),22.3(CH₃),12.7(CH₃),-2.9(CH₃),-3.3(CH₃) (ii) a Z-olefins (minor diastereomers), 163.3(C),152.3(C),138.4(C),138.0(CH),134.0(CH),132.7(C),129.3(CH),128.0(CH),127.2(C),124.6(C),120,6(CH),109.6(CH),53.8 (CH)₃),51.1(CH₂),43.3(CH₂),41.9(CH),39.7(C),34.7(CH),27.9(CH),22.0(CH₃),12.8(CH₃),-3.0(CH₃),-3.5(CH₃)。IR(ATR-FTIR),cm^-1:2952(br),1598(m),1578(w),1476(s),1426(m),1320(m),1264(m),1112(w),1031(w),824(m),733(w),702(w)。HRMS-CI(m/z):[M+H]⁺C₂₅H₃₁N₂OSi calculated value 403.2201; found 403.2198.

Analysis by NOE (500MHz, CDCl)₃) Indicating that the secondary diastereomer has the Z-configuration. See fig. 3.

Steps 3 a-d: conversion of the olefination product 10 to (-) -huperzine a (1):

step 3 a: yuba-fleming oxidation of olefination product 10 (alcohol 11):

triflic acid (2.29 mL, 26.0mmol, 2.20 equiv.) is added dropwise via syringe to a stirred solution of olefination product 10 (4.74 g, 11.8mmol, 1.0 equiv.) in dichloromethane (59mL) over 5 minutes at 0 ℃. The reaction mixture was warmed to 24 ℃ over 10 minutes. The reaction mixture was stirred at 24 ℃ for 1 h. The solvent was evaporated under reduced pressure.The resulting residue was dissolved in N, N-dimethylformamide (94mL) and potassium carbonate (4.89 g, 35.4mmol, 3.00 equiv.) and distilled water (47mL) were then added in that order the resulting milky solution was stirred at 24 ℃ for 15 minutes, a solution of tetrabutylammonium fluoride in tetrahydrofuran (1.0M, 177mL, 177mmol, 15.0 equiv.) was added, the resulting mixture was stirred at 24 ℃ for 1 hour then an aqueous solution of hydrogen peroxide (35%, 30.4mL, 354mmol, 30.0 equiv.) was added rapidly, the resulting mixture was warmed to 40 ℃ the reaction mixture was stirred and heated at 40 ℃ for 12 hours, the product mixture was cooled to 24 ℃ over 10 minutes, the cooled product mixture was transferred to a separatory funnel already filled with distilled water (300mL) and 50% ethyl acetate-hexane (v/v,500mL), the resulting layer was separated, the organic layer was washed with water (5 mL) (× mL) and saturated aqueous sodium chloride (2 mL) in that order, the organic layer was washed, dried, filtered, and the resulting solution was concentrated as a pale yellow, filtered, dried, 35.11 g, dried, and the alcohol was added.¹HNMR analysis (400MHz, CDCl)₃) Indicating conversion to alcohol 11>95 percent. The product thus obtained was used directly in the next step.

An analytically pure sample of alcohol 11 was obtained by flash column chromatography (eluting with 50% ethyl acetate-hexanes):

R_f=0.30 (50% ethyl acetate-hexanes, KMnO)₄）。¹H NMR（500MHz，CDCl₃5:1 mixtures of diastereomers); e-olefin (major diastereomer), 7.69(d,1H, J =8.5Hz, H₁),6.64(d,1H,J=8.5Hz,H₂),6.12(q,1H,J=6.5Hz,H₁₀),3.89(s,3H,H₃),3.54(dd,1H,J=6.0,3.5Hz,H₆),3.29-3.27(m,1H,H₅),3.10(dd,1H,J=18.5,6.5Hz,H₄),2.99(d,1H,J=17.5Hz,H₄),2.59(dd,1H,J=13.5,7.0Hz,H₈),1.79(d,3H,J=7.0Hz,H₁₁),1.87-1.76(m,2H,H₇/H₈),0.71(d,3H,7.5Hz,H₉) (ii) a Z-olefin (minor diastereomer), 7.78(d,1H, J-8.5Hz, H₁),6.67(d,1H,J=8.5Hz,H₂),5.65(q,1H,J=7.5Hz,H₁₀),3.90(s,3H,H₃),3.43(dd,1H,J=5.5,3.5Hz,H₆),3.17(dd,1H,J=18.0,7.0Hz,H₄),2.94(d,1H,J=18.0Hz,H₄),2.70(dd,1H,J=13.5,7.5Hz,H₈),2.62-2.60(m,1H,H₅),2.07(d,3H,J=7.0Hz,H₁₁),1.87-1.76(m,2H,H₇/H₈),0.68(d,3H,J=7.5Hz,H₉)。¹³C NMR（125MHz，CDCl₃5:1 mixtures of diastereomers); e-olefin (major diastereomer), 163.5(C),152.3(C),137.7(CH),131.5(C),126.4(C),122.0(C),120.4(CH),109.7(CH),78.4(CH),53.8 (CH)₃),44.7(CH₂),44.5(C),39.1(CH),37.9(CH₂),34.2(CH),17.9(CH₃),12.8(CH₃) (ii) a Z-olefins (minor diastereomers), 163.5(C),152.6(C),137.5(CH),129.8(C),126.0(C),122.8(CH),122.0(C),109.7(CH),77.9(CH),53.8 (CH)₃),49.3(CH),45.5(CH₂),44.5(C),37.9(CH₂),34.2(CH),17.9(CH₃),12.8(CH₃)。IR(ATR-FTIR),cm^-1:3431(br),2925(br),1598(m),1577(w),1476(s),1422(m),1323(m),1267(m),1033(m),828(w),658(w)。HRMS-CI(m/z):[M+H]⁺C₁₇H₂₁N₂O₂Calculated value 285.1598; found 285.1597.

And step 3 b: dehydration of the Jade tail-Flemine Oxidation product 11 (olefin 16):

to a 100mL round bottom flask connected to a teflon coated valve was added sequentially unpurified yurt-fleming oxidation product 11 (11.8 mmol, 1.00 eq., assuming quantitative yield in the previous step) and N- (triethylammoniumsulfonyl) carbamic acid methyl ester 12 (3.09 g, 13.0mmol, 1.10 eq.). Benzene (10mL) was added and the resulting solution was stirred at 24 ℃ for 15 minutes. The solution was concentrated to dryness and the resulting residue was dissolved in toluene (59 mL). The reaction vessel was sealed and the sealed vessel was then placed in an oil bath preheated to 110 ℃. The reaction mixture was stirred and heated at 110 ℃ for 12 h. The product mixture was cooled to 24 ℃ over 30 minutes. Will cool downThe product mixture of (2) was diluted with ethyl acetate (200mL) and the diluted solution was transferred to a separatory funnel already containing a saturated aqueous solution of sodium bicarbonate (200 mL). The formed layers are separated. The aqueous layer was extracted with ethyl acetate (200 mL). The organic layers were combined and the combined organic layers were dried over sodium sulfate. The dried solution was filtered and the filtrate was concentrated to give olefin 16(3.19g) as an off-white solid.¹H NMR analysis (400MHz, CDCl)₃) Indicating conversion to olefin 16>95 percent. The product thus obtained was used directly in the next step. An analytically pure sample of the olefin 16 was obtained by flash column chromatography (eluting with 10% ethyl acetate-hexanes):

R_f=0.32 (10% ethyl acetate-hexanes, KMnO)₄）。¹H NMR（400MHz，CDCl₃5:1 mixture of diastereomers): e-olefin (major diastereomer), 7.70(d,1H, J =8.8Hz, H₁),6.63(d,1H,J=8.8Hz,H₂),5.95(q,1H,J=6.8Hz,H₉),5.48(m,1H,H₆),3.89(s,3H,H₃),3.62(m,1H,H₅),2.98(dd,1H,J=17.2,5.2Hz,H₄),2.88-2.80(m,2H,H₄/H₇),2.38(d,1H,J=16.8Hz,H₇),1.76(d,3H,J=6.8Hz,H₁₀),1.55(s,3H,H₈) (ii) a Z-olefin (minor diastereomer), 7.78(d,1H, J =8.4Hz, H₁),6.66(d,1H,J=8.4Hz,H₂),5.65(q,1H,J=7.2Hz,H₉),5.46(d,1H,J=4.8Hz,H₆),3.89(s,3H,H₃),3.10-2.77(m,4H,2xH₄/H₅/H₇),2.38(d,1H,J=16.8Hz,H₇),2.06(d,3H,J=7.6Hz,H₁₀),1.54(s,3H,H₈)。¹³C NMR（100MHz，CDCl₃5:1 mixture of diastereomers): e-olefins (major diastereomer), 163.5(C),152.9(C),137.7(CH),132.3(C),130.7(C),125.2(CH),124.8(C),121.7(C),116.7(CH),109.2(CH),53.7 (CH)₃),47.5(CH₂),44.6(C),39.8(CH₂),31.6(CH),22.6(CH₃),12.7(CH₃) (ii) a Z-olefins (minor diastereomers), 163.5(C),153.2(C),137.7(CH),130.9(C),130.2(C),126.3(CH),124.6(C),121.7(C),119.0(CH),109.3(CH),53.7 (CH)₃),48.3(CH₂),42.1(CH),40.7(CH₂),40.1(C),22.5(CH₃),12.3(CH₃)。IR(ATR-FTIR),cm^-1:2934(br),1598(m),1576(w),1476(s),1421(m),1323(m),1268(m),1028(w),826(w)。HRMS-CI(m/z):[M+H]⁺C₁₇H₁₉N₂O calculated value 267.1492; found 267.1492.

And step 3 c: hydrolysis of nitrile 16 (amide 14):

hydrogenation of (Hydroxydimethylphosphine) [ Hydrobis (Hydroxydimethylphosphine)]Platinum (II) (13, 101mg, 240. mu. mol, 0.02 eq) was added to a solution of unpurified nitrile 16 (11.8 mmol, 1.00 eq, assuming quantitative yield in the previous step) in ethanol (6.6mL) and water (3.3 mL). The resulting mixture was placed in an oil bath preheated to 95 ℃. The reaction mixture was stirred and heated at 95 ℃ for 24 h. The product mixture was cooled to 24 ℃ over 10 minutes. The cooled mixture was concentrated to dryness. The resulting residue was dissolved in dichloromethane (15mL) and chloroform (15mL), and the resulting solution was filtered through a pad of sodium sulfate. The filtrate was concentrated to give amide 14(3.60g) as an off-white solid.¹H NMR analysis (400MHz, CDCl)₃) Indicating conversion to amide 14>95 percent. The product thus obtained was used directly in the next step. An analytically pure sample of amide 14 was obtained by flash column chromatography (eluting with 50% ethyl acetate-hexanes):

R_f=0.20 (50% ethyl acetate-hexanes, KMnO)₄）。¹H NMR（500MHz，CDCl₃5:1 mixture of diastereomers): e-olefin (major diastereomer), 7.33(d,1H, J =8.5Hz, H₁),6.57(d,1H,J-8.5Hz,H₂),5.62(brs,1H,H₁₁),5.40(q,1H,J=7.0Hz,H₉),5.38-5.35(m,1H,H₆),5.17(br s,1H,H₁₁),3.90(s,3H,H₃),3.60(m,1H,H₅),3.09-3.01(m,2H,H₄/H₇),2.88(d,1H,J=16.5Hz,H₄),2.11(d,1H,J=17.5Hz,H₇),1.70(d,3H,J=7.0Hz,H₁₀),1.53(s,3H,H₈) (ii) a Z-olefin (minor diastereomer), 7.37(d,1H, J =8.4Hz, H₁),6.58(d,1H,J=8.4Hz,H₂),5.58(br s,1H,H₁₁),5.54(q,1H,J=16.5Hz,H₉),5.38-5.35(m,1H,H₆),5.30(br s,1H,H₁₁),3.90(s,3H,H₃),3.15-3.01(m,3H,H₄/H₅/H₇),2.83(d,1H,J=16.5Hz,H₄),2.18(d,1H,J=17.0Hz,H₇),1.73(d,3H,J=7.5Hz,H₁₀),1.53(s,3H,H₈)；¹³CNMR（125MHz，CDCl₃5:1 mixture of diastereomers): e-olefin (major diastereomer), 176.9(C),162.9(C),153.8(C),138.9(CH),138.1(C),133.7(C),128.5(C),124.1(CH),115.3(CH),108.9(CH),54.4(C),53.7 (CH)₃),45.3(CH₂),39.8(CH₂),33.0(CH),23.0(CH₃),13.0(CH₃) (ii) a Z-olefins (minor diastereomers), 178.4(C),162.9(C),153.1(C),138.5(CH),137.1(C),133.6(C),128.3(C),125.9(CH),117.5(CH),109.2(CH),53.7 (CH)₃),51.2(C),45.1(CH₂),44.2(CH),39.7(CH₂),23.0(CH₃),13.0(CH₃)。IR(ATR-FTIR),cm^-1:HRMS-CI(m/z):3346(br),2926(br),1710(w),1664(s),1597(m),1576(w),1475(s),1422(m),1322(m),1267(w),1028(m),824(w)。[M+H]⁺C₁₇H₂₁N₂O₂Calculated value 285.1598; found 285.1601.

And step 3 d: conversion of amide 14 to (-) -huperzine a (1):

[ bis (trifluoroacetyloxy) iodo ] benzene (5.58 g, 13.0mmol, 1.10 equiv.) was added to a stirred solution of unpurified amide 14 (11.8 mmol, 1.00 equiv., assuming quantitative yield in the previous step) in methanol (240 mL). The resulting mixture was heated to reflux (bath temperature =65 ℃). The reaction mixture was stirred and heated at 65 ℃ for 2 h. The product mixture was cooled to 24 ℃ over 30 minutes. The cooled mixture was concentrated to dryness. The resulting residue was dissolved in chloroform (120 mL). Trimethylsilyl iodide (8.40 mL, 59.0mmol, 5.00 equiv) was added and the reaction mixture was heated to reflux (bath temperature =61 ℃). The reaction mixture was stirred and heated at 61 ℃ for 3 h. The mixture was then cooled to 24 ℃ over 30 minutes. Methanol (120mL) was added and the resulting mixture was heated to reflux (bath temperature =65 ℃). The reaction mixture was stirred and heated at 65 ℃ for 12 h. The product mixture was then cooled to 24 ℃ over 30 minutes. The cooled product mixture was concentrated to dryness. The resulting residue was dissolved in 50% dichloromethane-chloroform (v/v,200 mL). The resulting solution was transferred to a separatory funnel already filled with 1.0N aqueous sulfuric acid (200 mL). The formed layers are separated. The aqueous layer was then extracted with 50% dichloromethane-chloroform (v/v, 2X 200 mL). The organic layers were combined and discarded. The aqueous layer was basified with saturated aqueous ammonium hydroxide solution (100mL, final pH = 12-13). The basified aqueous layer was extracted with 50% dichloromethane-chloroform (v/v, 4X 200 mL). The organic layers were combined and the combined organic layers were dried over sodium sulfate. The dried solution was filtered and the filtrate was concentrated. The resulting residue was subjected to flash column chromatography (eluting with 10% methanol-ethyl acetate) to afford (-) -huperzine a (1, 1.61g, 56%, off-white solid) and the olefin isomer (isophytine a, 17, 310mg, 11%, off-white solid).

Synthetic (-) -huperzine A (1) is, in all respects¹H NMR、¹³C NMR, LC/MS retention time, IR, TLC solvent system (10% methanol-ethyl acetate, 5% methanol-dichloromethane +1% ammonium hydroxide) and optical rotation]All identical to the real samples.

(-) -huperzine A (1): r_f=0.15 (10% methanol-ethyl acetate, KMnO)₄）。t_R=0.91。[α]²⁰ _n-144(c0.23,CHCl₃) Reference [ α ]]²⁰ _n=-150(c0.12,CHCl₃)。¹⁰ ¹H NMR(500MHz,CDCl₃),13.25(br s,1H,H₃),7.88(d,1H,9.5Hz,H₁,6.37(d,1H,J=9.0Hz,H₂),5.46(q,1H,J=6.5Hz,H₉),5.38(d,1H,J=4.5Hz,H₆),3.59-3.55(m,1H,H₅),2.86(dd,1H,J=17.0,5.0,H₄),2.73(dd,1H,J=16.5,1.0Hz,H₄),2.12(app s,2H,H₇),1.88(br s,2H,H₁₁),1.64d,3H,J=6.5Hz,H₁₀),1.51(s,3H,H₈).¹³CNMR(125MHz,CDCl₃),165.5(C),143.3(C),142.4(C),140.3(CH),134.1(C),124.4(CH),122.8(C),117.1(CH),111.4(CH),54.4(C),49.2(CH₂),35.4(CH₂),33.0(CH),22.7(CH₃),12.5(CH₃)。IR(ATR-FTIR),cm^-1:3355(br),1644(s),1608(s),1552(m),1452(m),1121(m),837(m)。HRMS-CI(m/z):[M+H]⁺C₁₅H₁₉N₂O calculated value 243.1492; found 243.1493.

Isohuperzine a (17): r_f=0.15 (5% methanol-dichloromethane +1% ammonium hydroxide, KMnO₄）。[α]²⁰ _n=-121(c0.01,CHCl₃)。¹H NMR(400MHz,CDCl₃),13.10(br s,1H,H₃),7.86(d,1H,7=9.6Hz,H₁),6.42(d,1H,J=9.6Hz,H₂),5.41(q,1H,J=7.2Hz,H₉),5.37(br s,1H,H₆),3.00-2.88(m,2H,H₄/H₅),2.70(d,1H,J=16.0Hz,H₄),2.40(d,1H,J=16.8,H₇),2.05(d,1H,H₇),1.93(d,3H,J=7.2Hz,H₁₀),1.90(brs,2H,H₁₁),1.53(s,3H,H₈)。¹³C NMR(100MHz,CDCl₃),165.5(C),143.4(C),140.2(C),140.0(CH),133.7(C),125.4(CH),123.0(C),117.3(CH),115.7(CH),56.6(C),49.8(CH₂),44.0(CH),36.4(CH₂),22.6(CH₃),14.0(CH₃)。IR(ATR-FTIR),cm^-1:3380(br),2909(br),1653(s),1611(m),1551(m),1459(m),833(m),755(m),651(m)。HRMS-CI(m/z):[M+H]⁺C₁₅H₁₉N₂O calculated value 243.1492; found 243.1494.

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Claims

1. A process for preparing substantially pure (-) huperzine a having the formula:

comprising reacting an amide of the formula:

subjecting to a modified Hofmann reaction in an aqueous or alcoholic solvent and in the presence of bis (trifluoroacetoxyiodo) benzene to form an intermediate, fully deprotecting the intermediate to form (-) huperzine A, and optionally further purifying the (-) huperzine A, wherein the substantially pure (-) huperzine A comprises greater than 80% by weight of (-) huperzine A and less than 20% by weight of (+) huperzine A.

2. The process of claim 1, wherein the alcohol solvent is methanol and the (-) huperzine a is further purified by crystallization or flash column chromatography.

3. The method of claim 1 or 2, wherein the substantially pure (-) huperzine comprises greater than 99% by weight (-) huperzine a and less than 1% by weight (+) huperzine a.

4. The method of claim 1 or 2, wherein the substantially pure (-) huperzine a comprises greater than 90% by weight (-) huperzine a and less than 10% by weight (+) huperzine a.

5. The method of claim 1 or 2, wherein the substantially pure (-) huperzine a comprises greater than 95% by weight (-) huperzine a and less than 5% by weight (+) huperzine a.

6. The method of claim 1 or 2, wherein the substantially pure (-) huperzine a comprises greater than 99.9% by weight (-) huperzine a and less than 0.1% by weight (+) huperzine a.

7. A process for preparing substantially pure (-) huperzine a comprising:

(a) in a one-pot process, (R) -4-methyl-cyclohex-2-en-1-one is reacted with lithium dimethylphenylsilylcuprate in a conjugate addition reaction to form an initial enolate and the initial enolate is alkylated by 3-bromo-2- (bromomethyl) -6-methoxypyridine to form an addition-alkylation product having the formula:

(b) in a one-pot process, the addition-alkylation product is deprotonated by: reacting the addition-alkylation product with lithium bis (trimethylsilyl) amide or lithium diisopropylamide in an organic solvent to form an alpha-cyanoketone, subjecting the alpha-cyanoketone to palladium-catalyzed intramolecular enolate heteroarylation in the presence of a base to form a cyclized product, and stereoselective olefination of a ketone functional group of the cyclized product in the presence of a base and in an organic solvent in a Wittig olefination reaction to form an olefination product, wherein the stereoselective olefination of the cyclized product kinetically favors the formation of the olefination product in the form of the E-isomer, and wherein the olefination product has the formula:

(c) subjecting the olefination product to oxydisilylation by: (1) with boron trifluoride-acetic acid complex or a bronsted acid in an inert solvent, or (2) oxidation using a fleming-jade tail to form a cyanohydrin having the formula:

(d) in a one-pot process, the cyanohydrin is dehydrated in an organic solvent under heating and in the presence of a bergius reagent to form a dehydration product, and the dehydration product is subjected to pyrolysis in an alcohol and in the presence of a platinum catalyst to form an amide having the formula:

and

(f) subjecting the amide to a modified hofmann reaction in an aqueous or alcoholic solvent and in the presence of bis (trifluoroacetoxyiodo) benzene to form an intermediate, fully deprotecting the intermediate to form (-) huperzine a, and optionally further purifying the (-) huperzine a to obtain substantially pure (-) huperzine a having the formula:

wherein the substantially pure (-) huperzine A comprises greater than 80% by weight (-) huperzine A and less than 20% by weight (+) huperzine A.

8. The method of claim 7, wherein:

(a) reacting the addition-alkylation product with lithium bis (trimethylsilyl) amide or lithium diisopropylamide in THF or toluene;

(b) the palladium-catalyzed intramolecular enolate heteroarylated base is sodium tert-butoxide;

(c) the Wittig olefination reaction base is selected from the group consisting of n-butyllithium, sodium bis (trimethylsilyl) amide, lithium bis (trimethylsilyl) amide, potassium bis (trimethylsilyl) amide, and lithium diisopropylamide; and the Wittig olefination reaction organic solvent is selected from THF, diethyl ether or 1, 4-dioxane;

(d) the oxydimethylsilylated Bronsted acid is selected from TFA, MSA, FMSA or tetrafluoroboric acid;

(e) the disilylated inert solvent is DCM;

(f) the cyanohydrin dehydration organic solvent is toluene;

(g) the pyrolysis alcohol is hydrous ethanol;

(h) the improved Hofmann reaction alcohol solvent is methanol; and

(i) the (+/-) huperzine A is purified by flash column chromatography.

9. The method of claim 7 or 8, wherein the substantially pure (-) huperzine a contains less than one percent (+) huperzine a by weight.

10. The method of claim 7 or 8, wherein the substantially pure (-) huperzine a comprises greater than 90% by weight (-) huperzine a and less than 10% by weight (+) huperzine a.

11. The method of claim 7 or 8, wherein the substantially pure (-) huperzine a comprises greater than 95% by weight (-) huperzine a and less than 5% by weight (+) huperzine a.

12. The method of claim 7 or 8, wherein the substantially pure (-) huperzine a comprises greater than 99.9% by weight (-) huperzine a and less than 0.1% by weight (+) huperzine a.