Patent Description:
Sterol regulatory element-binding proteins (SREBPs) are one of families of transcription factors involved in lipid homeostasis. SREBPs control lipid metabolism in all tissues, by regulating expression of the genes related to biosynthesis and uptake of fatty acids, triglycerides, cholesterol, and phospholipids. Because of their central roles in lipid metabolism, SREBPs are strongly linked to metabolic syndromes. For example, high insulin levels, induced by high calorie diets or obesity, hyper-activate SREBPs, causing triglyceride accumulation and inducing fatty liver diseases. Hyper-activation of SREBPs also increases cholesterol levels and suppresses insulin receptor substrate-<NUM>, leading to hyperlipidemia, arteriosclerosis, and insulin resistance. Furthermore, activation of SREBPs is often correlated with the growth of cancers and the ability of hepatitis virus to cause fatty liver diseases (Non Patent Literature <NUM>). The involvement of SREBP activation in multiple diseases has made these transcription factors attractive pharmaceutical targets. To date, the only known "endogenous" molecules that directly inhibit the SREBP activation pathway are sterols. Further, reference is made in the literature to <NUM>,<NUM>-dihydroxyvitamin D<NUM>'s preventing effect against high fat diet-induced hepatic steatosis in rats as being related to the inhibition of lipogenesis and the promotion of fatty acid oxidation in rat liver (Non Patent Literature <NUM>), vitamin D deficiency as promoting nonalcoholic steatohepatitis through impaired enterohepatic circulation in an animal model (Non Patent Literature <NUM>), and the molecular mechanism of <NUM>,<NUM>-dihydroxyvitamin D3 inhibition of adipogenesis in 3T3-L1 cells (Non Patent Literature <NUM>).

It is an object of the present invention to provide compounds useful as a SREBP inhibitor and useful for treating a disease such as metabolic disease including non-alcoholic steatohepatitis (NASH); liver disease including fatty liver; diabetes; cancer; obesity; cardiovascular disease; and the like.

The inventors have found novel vitamin D<NUM> derivatives as a SREBP inhibitor. The vitamin D<NUM> derivatives in the present invention are useful for treating a disease such as metabolic disease including non-alcoholic steatohepatitis (NASH); liver disease including fatty liver; diabetes; cancer; obesity; cardiovascular disease; and the like.

In one aspect, the present invention is directed to a compound of the following general formula (I):
<CHM>
wherein one of RA and RB is hydroxyl and the other is NR<NUM>R<NUM>;
R<NUM> and R<NUM> are each independently selected from hydrogen; C<NUM>-<NUM> alkyl; C<NUM>-<NUM> alkylcarbonyl optionally substituted with at least one halogen which are the same or different; C<NUM>-<NUM> alkylsulfonyl; benzyloxycarbonyl; <NUM> to <NUM>-membered cycloalkyl-C<NUM>-<NUM> alkyl; C<NUM>-<NUM> arylcarbonyl optionally substituted with at least one group independently selected from the group consisting of halogen, halo-C<NUM>-<NUM> alkyl, -S-halo-C<NUM>-<NUM> alkyl, C<NUM>-<NUM> alkoxy, halo-C<NUM>-<NUM> alkoxy, nitro, cyano, C<NUM>-<NUM> alkoxycarbonyl and C<NUM>-<NUM> aryl; C<NUM>-<NUM> arylsulfonyl optionally substituted with at least one group independently selected from the group consisting of C<NUM>-<NUM> alkyl, nitro, and di-(C<NUM>-<NUM> alkyl)amino; <NUM> to <NUM>-membered saturated heterocyclyl-C<NUM>-<NUM> alkyl optionally substituted with at least one group independently selected from the group consisting of halogen and hydroxyl; <NUM> to <NUM>-membered heteroaryl; and a group of the following formula:
<CHM>
or.

provided that R<NUM> and R<NUM> are not concurrently hydrogen.

In another aspect, the present invention is also directed to the compound of Formula (I) or a pharmaceutically acceptable salt thereof for use in a method for treating a disease selected from metabolic disease, liver disease, obesity, diabetes, cardiovascular disease, or cancer in a subject.

In another aspect, the present invention is also directed to the compound of Formula (I) or a pharmaceutically acceptable salt thereof for use in a method for inhibiting SREBPs in a subject.

In still another aspect, the present invention is also directed to a pharmaceutical composition, comprising as the active ingredient the compound of Formula (I) or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

In still another aspect, the present invention is also directed to the compound of Formula (I) or a pharmaceutically acceptable salt thereof for use in the treatment of metabolic disease including non-alcoholic steatohepatitis; liver disease including fatty liver; diabetes; cancer; obesity; or cardiovascular disease.

In still another aspect, the present invention is also directed to the compound of Formula (I) or a pharmaceutically acceptable salt thereof for use in the treatment of obesity, non-alcoholic steatohepatitis (NASH); fatty liver; or cancer.

The compound of Formula (I) or a pharmaceutically acceptable salt thereof has SREBP inhibitory activity and may be useful for treating a disease such as metabolic disease including non-alcoholic steatohepatitis (NASH), liver disease including fatty liver; diabetes; cancer; obesity; cardiovascular disease or the like.

The term "alkyl" used herein refers to a straight- or branched-chain hydrocarbon group preferably having <NUM> to <NUM> carbon atoms, and includes, for example, methyl, ethyl, normal-propyl, isopropyl, normal-butyl, isobutyl, tert-butyl, etc..

The term "alkoxy" used herein refers to a monovalent group wherein the above mentioned alkyl group attaches to oxygen atom, and may be a straight- or branched-chain group preferably having <NUM> to <NUM> carbon atoms. The alkoxy group includes, for example, methoxy, ethoxy, normal-propoxy, isopropoxy, normal-butoxy, isobutoxy, tert-butoxy, etc..

The term "alkylcarbonyl" used herein refers to a group wherein the above mentioned alkyl group attaches to carbonyl group, and is preferably C<NUM>-<NUM> alkylcarbonyl. The alkylcarbonyl group includes, for example, acetyl, ethylcarbonyl, normal-propylcarbonyl, isopropylcarbonyl, normal-butylcarbonyl, isobutylcarbonyl, tert-butylcarbonyl, etc..

The term "alkylsulfonyl" used herein refers to a group wherein the above mentioned alkyl group attaches to sulfonyl group, and is preferably C<NUM>-<NUM> alkylsulfonyl. The alkylsulfonyl group includes, for example, methylsulfonyl, ethylsulfonyl, normal-propylsulfonyl, isopropylsulfonyl, normal-butylsulfonyl, isobutylsulfonyl, tert-butylsulfonyl, etc..

The term "alkoxycarbonyl" used herein refers to a group wherein the above mentioned alkoxy group attaches to carbonyl group, and is preferably C<NUM>-<NUM> alkoxycarbonyl. The alkoxycarbonyl group includes, for example, methoxycarbonyl, ethoxycarbonyl, normal-propoxycarbonyl, isopropoxycarbonyl, normal-butoxycarbonyl, isobutoxycarbonyl, tert-butoxycarbonyl, etc..

The term "cycloalkyl" used herein refers to a saturated aliphatic monocyclic hydrocarbon ring preferably having <NUM> to <NUM> carbon atoms. The cycloalkyl group includes, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, etc. A preferable cycloalkyl group is <NUM> to <NUM>-membered cycloalkyl, and more preferable one is cyclopropyl.

The term "aryl" used herein refers to a monovalent group of monocyclic aromatic hydrocarbon ring or polycyclic aromatic hydrocarbon ring preferably having <NUM> to <NUM> carbon atoms. The aryl group includes, for example, phenyl, naphthyl, etc. A preferable aryl is C<NUM>-<NUM> aryl, and more preferable one is phenyl or naphthyl.

The term "arylcarbonyl" used herein refers to a group wherein the above mentioned aryl group attaches to carbonyl group, and is preferably C<NUM>-<NUM> arylcarbonyl. The arylcarbonyl group includes, for example, benzoyl, naphthylcarbonyl, etc. A preferable arylcarbonyl includes benzoyl, etc..

The term "arylsulfonyl" used herein refers to a group wherein the above mentioned aryl group attaches to sulfonyl group, and is preferably C<NUM>-<NUM> arylsulfonyl. The arylsulfonyl group includes, for example, phenylsulfonyl, naphthylsulfonyl, etc. A preferable arylsulfonyl includes phenylsulfonyl, etc..

The term "heterocyclyl" or "heterocyclic" used herein refers to a monovalent group of saturated or partially unsaturated <NUM> to <NUM>-membered monocyclic group having at least one heteroatom, preferably one or two heteroatom(s), independently selected from nitrogen, oxygen or sulfur and carbon atoms. The heterocyclyl group includes, for example, pyrrolidinyl, oxazolinyl, pyrazolidinyl, piperidyl, piperazinyl, morpholinyl, etc. A preferable heterocyclyl group includes pyrrolidinyl, piperidyl, morpholinyl, etc..

The term "heteroaryl" used herein refers to a monovalent group of aromatic cyclic group having at least one heteroatom independently selected from nitrogen, oxygen or sulfur and carbon atoms, and is preferably <NUM> to <NUM>-membered heteroaryl group. The heteroaryl group includes, for example, pyrrolyl, isoxazolyl, isothiazolyl, pyrazolyl, oxazolyl, thiazolyl, imidazolyl, pyridyl, pyrimidinyl, etc. A preferable heteroaryl group includes thiazolyl, pyridyl, etc..

The term "nitrogen-containing oxo-substituted saturated <NUM> to <NUM>-membered heterocyclic ring which may be optionally fused with a C<NUM>-<NUM> aryl ring" used herein refers to the above mentioned heterocyclyl ring containing at least one nitrogen atom in the ring which is substituted with at least one oxo group, and includes, for example, γ-lactam, δ-lactam, phthalimidyl, etc..

The term "halogen" or "halo" used herein refers to fluorine atom, chlorine atom, bromine atom, iodine atom, etc..

The optionally substituted alkylcarbonyl refers to the above mentioned alkylcarbonyl which may be optionally substituted with the same or different at least one halogen. The substituent in the optionally substituted alkylcarbonyl includes the same or different <NUM> to <NUM>, preferably <NUM> to <NUM>, halogen atom(s) and in particular three fluorine atoms.

The optionally substituted arylcarbonyl refers to the above mentioned arylcarbonyl which may be optionally substituted with the same or different at least one group selected from the group consisting of halogen, halo-C<NUM>-<NUM> alkyl, -S-halo-C<NUM>-<NUM> alkyl, C<NUM>-<NUM> alkoxy, halo-C<NUM>-<NUM> alkoxy, nitro, cyano, C<NUM>-<NUM> alkoxycarbonyl, and C<NUM>-<NUM> aryl. Preferable substituents in the optionally substituted arylcarbonyl include the same or different <NUM> to <NUM> group(s) selected from the group consisting of chloro, fluoro, bromo, methyl, methoxy, trifluoromethyl, methoxycarbonyl, trifluoromethoxy, nitro, cyano, -S-CF<NUM>, phenyl, etc..

The optionally substituted arylsulfonyl refers to the above mentioned arylsulfonyl which may be optionally substituted with the same or different at least one group selected from the group consisting of C<NUM>-<NUM> alkyl and nitro. A preferable substituent in the optionally substituted arylsulfonyl includes methyl, nitro, etc..

The optionally substituted <NUM> to <NUM>-membered saturated heterocyclyl-alkyl refers to the above mentioned alkyl substituted with the above mentioned heterocyclyl which may be optionally substituted with the same or different at least one group selected from the group consisting of halogen and hydroxyl. A preferable substituent in the optionally substituted <NUM> to <NUM>-membered saturated heterocyclyl-alkyl includes fluoro, hydroxy, etc..

In one aspect, the present invention is directed to the following Items or embodiments.

One of RA and RB is hydroxyl and the other is NR<NUM>R<NUM>.

In one embodiment, R<NUM> and R<NUM> each independently include hydrogen, tert-butoxycarbonyl, benzyloxycarbonyl, acetyl, p-methylphenylsulfonyl, o-nitrophenylsulfonyl, p-trifluoromethylbenzoyl, p-bromobenzoyl, ethylcarbonyl, propylcarbonyl, p-methoxybenzoyl, p-fluorobenzoyl, p-[(trifluoromethyl)thio]benzoyl, <NUM>,<NUM>,<NUM>,<NUM>-tetrafluorobenzoyl, <NUM>,<NUM>,<NUM>-trifluorobenzoyl, <NUM>,<NUM>-dimethoxybenzoyl, <NUM>,<NUM>,<NUM>-trifluorobenzoyl, <NUM>,<NUM>-difluorobenzoyl, <NUM>,<NUM>-difluorobenzoyl, <NUM>-chloro-<NUM>-fluorobenzoyl, <NUM>-chloro-<NUM>-fluorobenzoyl, p-nitrobenzoyl, <NUM>-trifluoromethyl-<NUM>-fluorobenzoyl, <NUM>-trifluoromethyl-<NUM>-fluorobenzoyl, p-trifluoromethoxybenzoyl, p-cyanobenzoyl, p-methoxycarbonylbenzoyl, p-phenylbenzoyl, <NUM>-morpholinylethyl, <NUM>-(<NUM>-fluoropiperidinyl)ethyl, <NUM>-(<NUM>-hydroxypiperidinyl)ethyl, <NUM>-pyridyl, <NUM>-thiazolyl, cyclopropylmethyl, ethyl, butyl, methylsulfonyl, trifluoromethylcarbonyl, <NUM>-dimethylamino-<NUM>-naphthylsulfonyl, and a group of the following probe structure:
<CHM>
, etc..

In another embodiment, R<NUM> and R<NUM> may combine together with the nitrogen atom to which they attach to form for example γ-lactam, δ-lactam or phthalimidyl.

In still another embodiment, R<NUM> is =CH<NUM>.

In still another embodiment, R<NUM> is hydrogen.

The pharmaceutically acceptable salt used herein refers to any salts which are known in the art and do not have excess toxicity. In particular, the pharmaceutically acceptable salt may include a salt with an inorganic acid, an organic acid, an inorganic base, or an organic base. Such an inorganic acid includes hydrofluoric acid, hydrochloric acid, hydrobromic acid, hydroiodic acid, nitric acid, sulfuric acid, and phosphoric acid. Such an organic acid includes acetic acid, trifluoroacetic acid, benzoic acid, p-toluenesulfonic acid, citric acid, oxalic acid, maleic acid, fumaric acid, lactic acid, malic acid, succinic acid, and tartaric acid. Such an inorganic base includes lithium, sodium potassium, magnesium, calcium, aluminum, and zinc. Such an organic base includes arginine and lysine. A preferable pharmaceutically acceptable salt is a salt with an inorganic acid, and in particular hydrochloride.

The pharmaceutically acceptable carrier used herein includes various conventional organic or inorganic carrier substances, for example, substances in solid preparations such as excipients, disintegrators, binders, glidants and lubricants, commonly used in the art, and substances in liquid preparations such as solvents, solubilizing agents, suspending agents, isotonizing agents, buffers and soothing agents, commonly used in the art. Additives commonly used in the art such as preservatives, antioxidants, colorants, and edulcorants may be added to a pharmaceutical composition in the present invention, if needed.

The compound of Formula (I) may be orally or parenterally administered in a therapeutically effective amount to mammals such as mice, rats, hamsters, guinea pigs, rabbits, cats, dogs, pigs, cattle, horses, sheep, monkeys, and human. While the therapeutically effective amount of the compound of Formula (I) may vary depending on subjects, diseases, dosage forms, routes of administration, and the like, the therapeutically effective amount of the compound of Formula (I) generally ranges for example from about <NUM> through about <NUM> to about <NUM> through about <NUM> per day, which may be administered once or several times in a divided amount.

For the avoidance of doubt, it is confirmed that in the general description above, the proposal of general preferences and options in respect of different features of the compounds, methods, use, and compositions constitutes in the usual way the proposal of general combinations of those general preferences and options for the different features, insofar as they are combinable and compatible and are put forward in the same context.

A method for preparing the compound of Formula (I) or a pharmaceutically acceptable salt thereof is illustrated as below, but is not limited thereto. For example, the schemes as below show illustrative preparation methods for exemplary compounds in the present invention. Compounds obtained in each step may be isolated or purified by known methods including distillation, recrystallization, column chromatography, etc., if needed, and may be also used in the next step without isolation or purification.

The following abbreviations may be used herein for example:.

The compound of Formula (I) wherein R<NUM> is =CH<NUM> may be prepared according to the following procedure:
<CHM>
In the scheme, X" is halogen, and the other symbols have the same meanings as defined in Item <NUM>.

A compound of Formula [a1] may be coupled with a compound of Formula [a2] in the presence of a palladium catalyst such as tetrakis(triphenylphosphine)palladium (<NUM>) (i.e., Pd(PPh<NUM>)<NUM>) and a base such as triethylamine in a solvent such as toluene to give a compound of Formula [a3]. The reaction temperature may range from room temperature to about <NUM>, preferably about <NUM>.

Compound [a1] and Compound [a2] may be prepared according to any one of the methods of preparing intermediate compounds below.

A protecting group such as tert-butyldimethylsilyl and triethylsilyl groups in a compound of Formula [a3] may be deprotected by treatment with hydrogen fluoride with a base such as 3HF • Et<NUM>N and HF • pyridine in a solvent such as tetrahydrofuran to give a compound of Formula [a4]. The reaction temperature may be any temperature that the reaction can proceed, preferably room temperature.

A compound of Formula [a3] wherein R<NUM> and R<NUM> are not concurrently hydrogen (e.g. R<NUM> is arylsulfonyl and R<NUM> is hydrogen) may be optionally subjected to Mitsunobu reaction using an organic phosphine compound such as triphenylphosphine and an azocarboxylic acid ester such as diisopropyl azodicarboxylate in a solvent such as tetrahydrofuran. The resulting compound may be sequentially treated or a compound of Formula [a3] may be treated with a thiol such as <NUM>-dodecanethiol in the presence of a base such as sodium hydride in a solvent such as ether including diethylether to give a compound of Formula [a5]. The reaction temperature may range from <NUM> to room temperature, preferably room temperature or a gradually changed temperature starting from <NUM> and raising to room temperature.

A compound of Formula [a5] may be treated with R<NUM>X' wherein X' is halogen or hydroxyl in the presence of a base such as triethylamine in a solvent such as dichloromethane to give a compound of Formula [a3]. The reaction temperature may be any temperature that the reaction can proceed, preferably <NUM>. The resulting Compound [a3] may be then subjected to the deprotection according to Step 2a to give a compound of Formula [a4].

The compound of Formula (I) wherein R<NUM> is hydrogen or the comparative compound of formula [b6] may be prepared according to the following procedure:
<CHM>
In the scheme, RB1 is phthalimidyl or benzyloxy, RB2 is amino or hydroxyl, and the other symbols have the same meanings as defined in Item <NUM>.

A compound of Formula [b1] may be treated with a compound of Formula [b2] in the presence of a base such as lithium bis(trimethylsilyl)amide (i.e., LiHMDS) in a solvent such as tetrahydrofuran to give a compound of Formula [b3]. The reaction temperature may range from -<NUM> to room temperature, preferably a gradually changed temperature starting from -<NUM> and warming to room temperature.

Compound [b1] and Compound [b2] may be prepared according to any one of the methods of preparing intermediate compounds below.

A compound of Formula [b3] may be treated with a base such as hydrazine hydrate and potassium carbonate in a solvent such as methanol and ethanol to give a compound of Formula [b4]. The reaction temperature may range from room temperature to about <NUM>.

A compound of Formula [b4] may be treated with R<NUM>X' wherein X' is halogen or hydroxyl in the presence of a base such as triethylamine in a solvent such as dichloromethane, followed by the deprotection according to Step 2a to give a compound of Formula [b5]. The temperature of the reaction with R<NUM>X' may be any temperature that the reaction can proceed, preferably <NUM>.

A compound of Formula [b4] may be alternatively treated with a fluorination agent such as N,N-diethylaminosulfur trifluoride (i.e., DAST) in a solvent such as dichloromethane, followed by the deprotection according to Step 2a to give a compound of Formula [b6]. The temperature of the fluorination reaction may be any temperature that the reaction can proceed, preferably -<NUM>.

Preparation methods for intermediate Compounds 14a and 14b
<CHM>
In the scheme, R<NUM> has the same meaning as defined in Item <NUM> and X' is halogen or hydroxyl.

Preparation methods for intermediate Compound <NUM>
<CHM>
In the scheme, Compound <NUM> may be prepared from Compound <NUM> according to a common procedure in the art such as the method described in <NPL>.

In particular, methods of preparing the compounds of Formula (I) wherein R<NUM> is =CH<NUM> in the present invention are illustrated in the following schemes using the above prepared intermediate compounds or derivatives thereof which may be prepared in a similar way to the above schemes.

Preparation methods for Comparative Compounds 18a, 18b, 19a and 19b
<CHM>.

Derivatives of Comparative Compounds 18a and 18b may be also prepared in a similar procedure to the above. <CHM>
<CHM>
In the scheme, R<NUM> has the same meaning as defined in Item <NUM>.

Preparation methods for Compounds <NUM>, <NUM>, <NUM>, and <NUM>
<CHM>
<CHM>
In the scheme, R<NUM> has the same meaning as defined in Item <NUM> and X' is halogen or hydroxyl.

Alternatively, the compound of Formula (I) wherein R<NUM> is a group of the following formula:
<CHM>
in the present invention may be prepared according to the following scheme.

Preparation methods for Compound <NUM>
<CHM>.

In addition, methods of preparing the compounds of Formula (I) wherein R<NUM> is hydrogen in the present invention are illustrated in the following schemes.

Preparation methods for intermediate Compound <NUM>
<CHM>.

Preparation methods for intermediate Compounds <NUM> and <NUM>
<CHM>
<CHM>
<CHM>
<CHM>.

Preparation methods for Compounds <NUM> to <NUM> and derivatives thereof
<CHM>
In the scheme, R<NUM> has the same meaning as defined in Item <NUM> and X' is halogen or hydroxyl.

Preparation methods for Compounds <NUM>, <NUM> and Comparative Compound <NUM> and derivatives thereof
<CHM>
In the scheme, R<NUM> has the same meaning as defined in Item <NUM> and X' is halogen or hydroxyl.

Preparation methods for Comparative Compound <NUM>
<CHM>.

Unless otherwise stated, preparations were performed under an argon atmosphere using freshly dried solvents. All preparations were monitored by thin-layer chromatography using Merck silica gel <NUM> F<NUM> pre-coated plates (<NUM>) and were visualized by UV and p-anisaldehyde staining. Flash column chromatography was performed under pressurization using silica gel (particle size <NUM>-<NUM>) purchased from Cica or NH silica gel (NH-DM1020) purchased from FUJI SILYSIA CHEMICAL LTD. <NUM>H NMR spectra were recorded on JNM-ECX <NUM> or JNM-AL <NUM>. The spectra are referenced internally according to residual solvent signals of CDCl<NUM> (<NUM>H NMR; δ = <NUM> ppm) or CD<NUM>OD (<NUM>H NMR; δ = <NUM> ppm). Data for <NUM>H NMR spectra are reported as follows: chemical shift (δ ppm) (multiplicity, coupling constant (Hz), integration). Multiplicity and qualifier abbreviations are as follows: s = singlet, d = doublet, t = triplet, q = quartet, m = multiplet, br = broad. Mass spectra were recorded on JEOL JMS-T100X spectrometer with ESI-MS mode using methanol as solvent or JEOL JMS-MS700V spectrometer with FAB-MS mode using DMSO as solvent.

To a solution of BH<NUM>-SMe<NUM> (<NUM>, <NUM> mol) in THF (<NUM>) was added a solution of L-malic acid (<NUM>, <NUM> mol; purchased from Tokyo Chemical Industry Co. ) in THF (<NUM>) dropwise at <NUM>, then the reaction mixture was warmed to room temperature. After stirring for <NUM>, the reaction mixture was cooled to <NUM> before adding MeOH (<NUM>). After additive <NUM> at room temperature, the mixture was evaporated. Moreover, the residue was evaporated with MeOH (<NUM>) six times and acetone (<NUM>) two times.

To a crude triol <NUM> in acetone (<NUM>) was added p-TsOH • H<NUM>O (<NUM>). After stirring for <NUM>, Et<NUM>N (<NUM>) was added. After additive <NUM>, the mixture was evaporated. The residue was chromatographed on silica gel (hexane/ethyl acetate; <NUM>:<NUM> to <NUM>:<NUM>) to give acetal <NUM> (<NUM>, <NUM>%, <NUM> steps) as a colorless oil.

A solution of oxalyl chloride (<NUM>, <NUM> mmol) in CH<NUM>Cl<NUM> (<NUM>) was cooled to - <NUM> and added dry DMSO (<NUM>, <NUM> mmol) dropwise. After stirring for <NUM> at same temperature, a solution of acetal <NUM> (<NUM>, <NUM> mmol) in CH<NUM>Cl<NUM> (<NUM>) was added dropwise, and the reaction mixture was stirred for <NUM>. To a suspension was added Et<NUM>N (<NUM>, <NUM> mmol) dropwise, then the reaction mixture was warmed to room temperature. After additive <NUM>, ethyl(triphenylphosphoranylidene)acetate (<NUM>, <NUM> mmol) was added and stirred for <NUM> day. The reaction was quenched by H<NUM>O, and the aqueous layer was extracted with CH<NUM>Cl<NUM> three times. The combined mixture was dried over MgSO<NUM> and concentrated in vacuo. The residue was chromatographed on silica gel (hexane/ethyl acetate; <NUM>:<NUM> to <NUM>:<NUM>) to give ethyl ester <NUM> (<NUM>, <NUM>%) as yellow oil.

A solution of ethyl ester <NUM> (<NUM>, <NUM> mmol) in CH<NUM>Cl<NUM> (<NUM>) was added diisobutyl aluminum hydride in hexane (<NUM>, <NUM> mmol, <NUM>). After stirring for <NUM>, the reaction was quenched by careful addition of MeOH (<NUM>). The mixture was added sat. Rochelle salt aq. (<NUM>) and stirred for <NUM>. The organic layer was washed with sat. Rochelle salt aq. three times, and the aqueous layer was extracted with CHCl<NUM> three times. The combined mixture was dried over MgSO<NUM> and concentrated in vacuo. The residue was chromatographed on silica gel (hexane/ethyl acetate; <NUM>:<NUM> to <NUM>:<NUM>) to give alcohol <NUM> (<NUM>, <NUM>%) as a colorless oil.

To a solution of allyl alcohol <NUM> (<NUM>, <NUM> mmol) in pyridine (<NUM>) was added benzoyl chloride (<NUM>, <NUM> mmol) dropwise at <NUM>, then the reaction mixture was warmed to room temperature. After stirring for <NUM>, the reaction mixture was added H<NUM>O. The aqueous layer was extracted with ethyl acetate three times and organic layer was washed with brine. The combined mixture was dried over MgSO<NUM> and concentrated in vacuo. The residue was chromatographed on silica gel (hexane/ethyl acetate; <NUM>:<NUM>) to give benzoate ester (<NUM>, <NUM>%) as colorless oil.

To a benzoate ester <NUM> (<NUM>, <NUM> mmol) was added acetic acid (<NUM>) and H<NUM>O (<NUM>). After stirring at room temperature for <NUM>, the reaction mixture was evaporated. The residue was chromatographed on silica gel (hexane/ethyl acetate; <NUM>:<NUM>) to give diol <NUM> (<NUM>, <NUM>%) as a colorless oil.

A solution of diol <NUM> (<NUM>, <NUM> mmol) in pyridine (<NUM>) was addedp-toluenesulfonyl chloride (<NUM>, <NUM> mmol) at <NUM>, then the reaction mixture was slowly warmed to room temperature. After stirring for <NUM>, the reaction was quenched by H<NUM>O. The aqueous layer was extracted with ethyl acetate three times. The combined organic extracts were washed with brine, dried over MgSO<NUM>, and concentrated in vacuo. The residue was chromatographed on silica gel (hexane/ethyl acetate; <NUM>:<NUM> to <NUM>:<NUM>) to give tosylate <NUM> (<NUM>, <NUM>%) as a colorless oil.

A solution of tosylate <NUM> (<NUM>, <NUM> mmol) in THF (<NUM>) was added NaH (<NUM>, <NUM> mmol, <NUM>%) at <NUM>, then the reaction mixture was warmed to room temperature. After stirring for <NUM>, the reaction was quenched by sat. The aqueous layer was extracted with ethyl acetate three times. The combined organic extracts were washed with brine, dried over MgSO<NUM>, and concentrated in vacuo. The residue was chromatographed on silica gel (hexane/ethyl acetate; <NUM>:<NUM> to <NUM>:<NUM>) to give epoxide <NUM> (<NUM>, <NUM>%) as colorless oil.

A solution of trimethylsilylacetylene (<NUM>, <NUM> mmol) in THF (<NUM>) was added n-BuLi in hexane (<NUM>, <NUM> mmol, <NUM>) dropwise at -<NUM> and stirred for <NUM>. A solution of epoxide <NUM> (<NUM>, <NUM> mmol) in THF (<NUM>) and BF<NUM>-OEt<NUM> (<NUM>, <NUM> mmol) was added dropwise at the same temperature, then the reaction mixture was warmed to room temperature over <NUM>. The reaction was quenched with sat. NH<NUM>Cl aq. , the aqueous layer was extracted with ethyl acetate three times. The combined organic extracts were washed with brine, dried over MgSO<NUM>, and concentrated in vacuo. The residue was chromatographed on silica gel (hexane/ethyl acetate; <NUM>:<NUM> to <NUM>:<NUM>) to give alcohol <NUM> (<NUM>, <NUM>%) as a colorless oil.

Alcohol <NUM> (<NUM>) was added imidazole (<NUM>, <NUM> mmol) and tert-butyldimethylsilyl chloride (<NUM>, <NUM> mmol) at room temperature, then DMF was added until the reagents were dissolved. After stirring for <NUM>, the reaction mixture was diluted with diethyl ether and quenched with sat. NaHCO<NUM> aq. The aqueous layer was extracted with diethyl ether three times. The combined organic extracts were washed with H<NUM>O and brine, dried over MgSO<NUM>, and concentrated in vacuo. The residue was chromatographed on silica gel (hexane/ethyl acetate; <NUM>:<NUM> to <NUM>:<NUM>) to give silyl ether <NUM> (<NUM>, <NUM>%) as a colorless oil.

A solution of allyl alcohol <NUM> (<NUM>, <NUM> mmol) in MeOH (<NUM>) was added potassium carbonate (<NUM>, <NUM> mmol) at <NUM>, then the reaction mixture was cooled to room temperature. After stirring for <NUM>, the reaction was quenched by H<NUM>O. The aqueous layer was extracted with ethyl acetate three times. The combined organic extracts were washed with brine, dried over MgSO<NUM>, and concentrated in vacuo. The residue was chromatographed on silica gel (hexane/ethyl acetate; <NUM>:<NUM> to <NUM>:<NUM>) to give allyl alcohol <NUM> (<NUM>, <NUM>%) as colorless oil. <NUM>H NMR (<NUM>, CDCl<NUM>) δ <NUM>-<NUM> (m, <NUM>), <NUM> (d, J = <NUM>, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM> (dd, J = <NUM>, <NUM>, <NUM>), <NUM> (t, J = <NUM>, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>).

A solution of allyl alcohol <NUM> (<NUM>, <NUM> mmol) in CH<NUM>Cl<NUM> (<NUM>) was added trichloroacetyl isocyanate (<NUM>, <NUM> mmol) at <NUM>. The reaction was monitored by thin layer chromatography on silica gel plates (eluent: <NUM>:<NUM> ethyl acetate to hexane). After <NUM>, the starting material spot completely converted to a new spot, then the reaction mixture was concentrated in vacuo. A solution of the residue in MeOH (<NUM>) was added H<NUM>O (<NUM>) and potassium carbonate (<NUM>, <NUM> mmol) at <NUM>, then the reaction mixture was warmed to room temperature. After stirring for <NUM>, the reaction mixture was added H<NUM>O, the aqueous layer was extracted with CH<NUM>Cl<NUM> three times, dried over MgSO<NUM>, and concentrated in vacuo. The residue was filtered through a pad of silica gel to give carbamate <NUM>.

To a solution of carbamate <NUM> (<NUM> mmol) and Et<NUM>N (<NUM>, <NUM> mmol) in CH<NUM>Cl<NUM> (<NUM>) was added trifluoroacetic anhydride (<NUM>, <NUM> mmol) dropwise at <NUM>. After stirring for <NUM>, the organic layer was washed with H<NUM>O and brine. The combined organic extracts were dried over Na<NUM>SO<NUM>, and concentrated in vacuo to crude isocyanate <NUM>.

A suspension of t-BuOK (<NUM>, <NUM> mmol) and t-BuOH (<NUM>, <NUM> mmol) in THF (<NUM>) was cooled to <NUM>. A crude isocyanate <NUM> (<NUM> mmol) in THF (<NUM>) was slowly added dropwise. The reaction was monitored by thin layer chromatography on silica gel plates (eluent: <NUM>:<NUM> ethyl acetate to hexane). After stirring for <NUM>, the starting material spot completely converted to new spots. The reaction mixture was added acetic acid (<NUM>, <NUM> mmol) and H<NUM>O (<NUM>), then Boc<NUM>O was added until disappeared amine. After stirring for <NUM>, the amine completely converted to product, then the reaction mixture was diluted with ethyl acetate, the aqueous layer was extracted with ethyl acetate three times. The combined organic extracts were washed with brine, dried over MgSO<NUM>, and concentrated in vacuo. The residue was chromatographed on silica gel (hexane/ethyl acetate; <NUM>:<NUM> to <NUM>:<NUM>) to give Boc amine <NUM> as a diastereomeric mixture wherein the ratio of 14a : 14b was <NUM> : <NUM> determined by <NUM>H NMR.

To a solution of diastereomeric mixture <NUM> (<NUM> mmol) in THF (<NUM>) was added tetrabutylammonium fluoride in THF (<NUM>, <NUM>). After stirring for <NUM>, the reaction mixture was evaporated. The residue was chromatographed on silica gel (hexane/ethyl acetate; <NUM>:<NUM> to <NUM>:<NUM>) to give alcohol 15a (<NUM>, <NUM>%) and alcohol 15b (<NUM>, <NUM>%) as a single diastereomer.

To the alcohol 15a (<NUM>, <NUM> mmol) was added imidazole (<NUM>, <NUM> mmol) and tert-butyldimethylsilyl chloride (<NUM>, <NUM> mmol) at room temperature, then DMF was added until the reagents were dissolved. After stirring for <NUM>, the reaction mixture was diluted with diethyl ether and quenched with sat. NaHCO<NUM> aq. The aqueous layer was extracted with diethyl ether three times. The combined organic extracts were washed with H<NUM>O and brine, dried over MgSO<NUM>, and concentrated in vacuo. The residue was chromatographed on silica gel (hexane/ethyl acetate; <NUM>:<NUM>) to give 14a (<NUM>, q. ) as colorless oil. <NUM>H NMR (<NUM>, CDCl<NUM>) δ <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM> (br, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM> (t, J = <NUM>, <NUM>), <NUM>-<NUM> (br, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>).

By following the same procedure described for 14a, 14b (<NUM>, q. ) was obtained from 15b (<NUM>, <NUM> mmol) as colorless oil. <NUM>H NMR (<NUM>, CDCl<NUM>) δ <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM> (br, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM> (t, J = <NUM>, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM> (s, <NUM>), <NUM> (d, J = <NUM>, <NUM>), <NUM> (s, <NUM>), <NUM> (d, J = <NUM>, <NUM>).

Acetyl chloride (<NUM>, <NUM> mmol) was slowly added to EtOH (<NUM>) dropwise at <NUM>. After stirring for <NUM>, this solution was added to alcohol <NUM> (<NUM>, <NUM> mmol). After additional stirring for <NUM>, the reaction mixture was evaporated. The residue was washed with Et<NUM>O, affording amine as a yellow solid. To a solution of above amine (<NUM> mmol) in H<NUM>O (<NUM>) was added acetic acid (<NUM>, <NUM> mmol), DMT-MM (<NUM>, <NUM> mmol), N-methylmorpholine (<NUM>, <NUM> mmol) and stirred for <NUM>. The aqueous layer was extracted with ethyl acetate three times. The combined organic extracts were washed with brine, dried over MgSO<NUM>, and concentrated in vacuo. The residue was chromatographed on silica gel (hexane/ethyl acetate; <NUM>:<NUM> to <NUM>:<NUM>) to give acetamide α and β as a single diastereomer. To α-acetamide (about <NUM> mmol) was added imidazole (<NUM>, <NUM> mmol) and tert-butyldimethylsilyl chloride (<NUM>, <NUM> mmol) at room temperature, and then DMF was added until the reagents were dissolved. After stirring for <NUM>, the reaction mixture was diluted with diethyl ether and quenched with sat. NaHCO<NUM>. The aqueous layer was extracted with diethyl ether three times. The combined organic extracts were washed with H<NUM>O and brine, dried over MgSO<NUM>, and concentrated in vacuo. The residue was chromatographed on silica gel (hexane/ethyl acetate; <NUM>:<NUM> to <NUM>:<NUM>) to give 20a (<NUM>, <NUM>%, <NUM> steps) as a colorless oil.

By following the same procedure described for 20a, 20b (<NUM>, <NUM>%, <NUM> steps) was obtained from β-acetamide (about <NUM> mmol) as a colorless oil.

A solution of alcohol 15a (<NUM>, <NUM> mmol) in pyridine (<NUM>) was added acetic anhydride (<NUM>) at room temperature. After stirring for <NUM>, the reaction mixture was evaporated. The residue was chromatographed on silica gel (hexane/ethyl acetate; <NUM>:<NUM> to <NUM>:<NUM>) to give acetate ester <NUM> (<NUM>, <NUM>%).

Acetyl chloride (<NUM>, <NUM> mmol) was slowly added to EtOH (<NUM>) dropwise at <NUM>. After stirring for <NUM>, this solution was added to acetate ester <NUM> (<NUM>, <NUM> mmol). After additional stirring for <NUM>, the reaction mixture was evaporated. The residue was washed with Et<NUM>O, affording amine <NUM> as a yellow solid.

To a solution of crude amine <NUM> (<NUM> mmol) in CH<NUM>Cl<NUM> (<NUM>) was added Et<NUM>N (<NUM>, <NUM> mmol), o-nitrobenzenesulfonyl chloride (<NUM>, <NUM> mmol) at <NUM>, then the reaction mixture was warmed to room temperature. After stirring for <NUM>, the reaction was quenched by H<NUM>O. The aqueous layer was extracted with ethyl acetate three times. The combined organic extracts were washed with brine, dried over MgSO<NUM>, and concentrated in vacuo. The residue was filtered through a pad of silica gel to give acetate ester as colorless oil. To a solution of acetate ester (<NUM> mmol) in MeOH (<NUM>) was potassium carbonate (<NUM>, <NUM> mmol) at <NUM>, then the reaction mixture was warmed to room temperature. After stirring for <NUM>, H<NUM>O was added, and the aqueous layer was extracted with ethyl acetate three times. The combined organic extracts were washed with brine, dried over MgSO<NUM>, and concentrated in vacuo. The residue was chromatographed on silica gel (hexane/ethyl acetate; <NUM>:<NUM> to <NUM>:<NUM>) to give nosylate <NUM> (<NUM>, <NUM>%) as a colorless oil.

To the nosylate <NUM> (<NUM>, <NUM> mmol) was added imidazole (<NUM>, <NUM> mmol) and tert-butyldimethylsilyl chloride (<NUM>, <NUM> mmol) at room temperature, then DMF was added until the reagents were dissolved. After stirring for <NUM>, the reaction mixture was diluted with diethyl ether and quenched with sat. NaHCO<NUM> aq. The aqueous layer was extracted with diethyl ether three times. The combined organic extracts were washed with H<NUM>O and brine, dried over MgSO<NUM>, and concentrated in vacuo. The residue was chromatographed on silica gel (hexane/ethyl acetate; <NUM>:<NUM> to <NUM>:<NUM>) to give <NUM> (<NUM>, <NUM>%) as a colorless oil. <NUM>H NMR (<NUM>, CDCl<NUM>) δ <NUM>-<NUM> (m, <NUM>), <NUM> (d, J = <NUM>, <NUM>), <NUM> (ddd, J = <NUM>, <NUM>, <NUM>, <NUM>), <NUM> (dd, J = <NUM>, <NUM>, <NUM>), <NUM> (dd, J = <NUM>, <NUM>, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM> (ddd, J = <NUM>, <NUM>, <NUM>, <NUM>), <NUM> (ddd, J = <NUM>, <NUM>, <NUM>, <NUM>), <NUM> (t, J = <NUM>, <NUM>), <NUM> (ddd, J = <NUM>, <NUM>, <NUM>, <NUM>), <NUM> (ddd, J= <NUM>, <NUM>, <NUM>, <NUM>), <NUM> (s, <NUM>), <NUM> (s,<NUM>), <NUM> (s, <NUM>).

To a solution of crude amine <NUM> (<NUM>, <NUM> mmol) in CH<NUM>Cl<NUM> (<NUM>) was added Et<NUM>N (<NUM>, <NUM> mmol), p-toluenesulfonyl chloride (<NUM>, <NUM> mmol) at <NUM>, then the reaction mixture was warmed to room temperature. After stirring for <NUM>, the reaction was quenched by H<NUM>O. The aqueous layer was extracted with ethyl acetate three times. The combined organic extracts were washed with brine, dried over MgSO<NUM>, and concentrated in vacuo. The residue was filtered through a pad of silica gel to give acetate ester (<NUM>, not pure) as colorless oil. To a solution of acetate ester (<NUM> mmol) in MeOH (<NUM>) was potassium carbonate (<NUM>, <NUM> mmol) at <NUM>, then the reaction mixture was warmed to room temperature. After stirring for <NUM>, H<NUM>O was added, and the aqueous layer was extracted with ethyl acetate three times. The combined organic extracts were washed with brine, dried over MgSO<NUM>, and concentrated in vacuo. The residue was chromatographed on silica gel (hexane/ethyl acetate; <NUM>:<NUM> to <NUM>:<NUM>) to give tosylate <NUM> (<NUM>, <NUM>%) as a colorless oil.

To the tosylate <NUM> (<NUM>, <NUM> mmol) was added imidazole (<NUM>, <NUM> mmol) and tert-butyldimethylsilyl chloride (<NUM>, <NUM> mmol) at room temperature, then DMF was added until the reagents were dissolved. After stirring for <NUM>, the reaction mixture was diluted with diethyl ether and quenched with sat. NaHCO<NUM> aq. The aqueous layer was extracted with diethyl ether three times. The combined organic extracts were washed with H<NUM>O and brine, dried over MgSO<NUM>, and concentrated in vacuo. The residue was chromatographed on silica gel (hexane/ethyl acetate; <NUM>:<NUM> to <NUM>:<NUM>) to give <NUM> (<NUM>, <NUM>%) as a colorless oil. <NUM>H NMR (<NUM>, CDCl<NUM>) δ <NUM>-<NUM> (m, <NUM>), <NUM> (d, J = <NUM>, <NUM>), <NUM> (ddd, J = <NUM>, <NUM>, <NUM>, <NUM>), <NUM> (dd, J = <NUM>, <NUM>, <NUM>), <NUM> (dd, J = <NUM>, <NUM>, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM> (s, <NUM>), <NUM> (ddd, J = <NUM>, <NUM>, <NUM>, <NUM>), <NUM> (ddd, J = <NUM>, <NUM>, <NUM>, <NUM>), <NUM> (t, J = <NUM>, <NUM>), <NUM>- <NUM> (m, <NUM>), <NUM> (s, <NUM>), <NUM> (s,<NUM>), <NUM> (s, <NUM>).

To a solution of crude amine <NUM> (<NUM>, <NUM> mmol) in CH<NUM>Cl<NUM> (<NUM>) was added Et<NUM>N (<NUM>, <NUM> mmol), benzyl chloroformate (<NUM>, <NUM> mmol) at <NUM>, then the reaction mixture was warmed to room temperature. After stirring for <NUM>, the reaction was quenched by H<NUM>O. The aqueous layer was extracted with ethyl acetate three times. The combined organic extracts were washed with brine, dried over MgSO<NUM>, and concentrated in vacuo. The residue was chromatographed on silica gel (hexane/ethyl acetate; <NUM>:<NUM> to <NUM>:<NUM>) to give NHCbz <NUM> (<NUM>, <NUM>%) as a colorless oil.

To a solution of NHCbz <NUM> (<NUM>, <NUM> mmol) in CH<NUM>Cl<NUM> (<NUM>) was added <NUM>,<NUM>-lutidine (<NUM>, <NUM> mmol) and tert-butyldimethylsilyl trifluoromethanesulfonate (<NUM>, <NUM> mmol) at room temperature. After stirring for <NUM>, the reaction was quenched by sat. NaHCO<NUM> aq. The aqueous layer was extracted with CH<NUM>Cl<NUM> three times. The combined organic extracts were dried over MgSO<NUM>, and concentrated in vacuo. The residue was chromatographed on silica gel (hexane/ethyl acetate; <NUM>:<NUM> to <NUM>:<NUM>) to give <NUM> (<NUM>, <NUM>%) as a colorless oil.

Imidazole (<NUM>, <NUM> mmol), TBSCI (<NUM>µL, <NUM> mmol) were successively added to a solution of <NUM>-hydroxy piperidine (<NUM>, <NUM> mmol) in dichloromethane (<NUM>) at room temperature and stirred for <NUM> d. The reaction mixture was washed with H<NUM>O, sat. NaHCO<NUM> aq. and brine, dried over MgSO<NUM>, and concentrated in vacuo. The residue was evapolated with toluene for removing residual reagents to give crude product (<NUM>, not pure). To a solution of above crude product (<NUM>) in acetonitrile (<NUM>) was added <NUM>-bromoethanol (<NUM>, <NUM> mmol) and potassium carbonate (<NUM>, <NUM> mmol) at room temperature and then the mixture was heated under reflux for <NUM>. The reaction mixture was filtered through a pad of celite and the resulting filtrate was concentrated. Purification by flash chromatography on silica gel (NH silica gel, hexane/ethyl acetate; <NUM>:<NUM> to <NUM>:<NUM>) to give <NUM> (<NUM>, <NUM>%).

To a solution of vitamin D<NUM> (<NUM>, <NUM> mmol; purchased from Tokyo Chemical Industry Co. ) in dichloromethane (<NUM>) was added methanol (<NUM>) and the solution was cooled to - <NUM>. Ozone was passed through the solution at - <NUM> for <NUM>. After flushing nitrogen to the solution to remove the residual ozone, the resulting mixture was treated with NaBH<NUM> (<NUM>, <NUM> mmol) and stirred for <NUM> at - <NUM>, and then allowed to warm to room temperature. The reaction was quenched by the addition of <NUM> aq. HCl, and extracted with dichloromethane. The organic phase was washed with water and dried over MgSO<NUM>, and concentrated in vacuo. The residue was chromatographed on silica gel (hexane/ethyl acetate; <NUM>:<NUM> to <NUM>:<NUM>) to give diol <NUM> (<NUM>, ><NUM>%). <NUM>H NMR (CDCl<NUM>) δ <NUM> (d, J = <NUM>, <NUM>), <NUM> (dd, J = <NUM>, <NUM>, <NUM>), <NUM> (dd, J = <NUM>, <NUM>, <NUM>), <NUM> (dd, J = <NUM>, <NUM>, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m. <NUM>), <NUM>-<NUM> (m. <NUM>), <NUM> (d, J = <NUM>, <NUM>), <NUM> (s, <NUM>).

To a solution of diol <NUM> (<NUM>, <NUM> mmol) in THF (<NUM>) was successively added imidazole (<NUM>, <NUM> mmol), triphenylphosphine (<NUM>, <NUM> mmol), iodide (<NUM>, <NUM> mmol) at - <NUM> and stirring for <NUM>. The reaction mixture was warmed to room temperature. After additional stirring for <NUM>, the mixture was cooled to <NUM> before adding sat. NaHCO<NUM> aq. The aqueous layer was extracted with ethyl acetate three times. The combined organic extracts were washed with sat. Na<NUM>S<NUM>O<NUM> aq. and H<NUM>O, dried over MgSO<NUM>, and concentrated in vacuo. The residue was chromatographed on silica gel (hexane/ethyl acetate; <NUM>:<NUM>) to give iodide <NUM> (<NUM>, <NUM>%).

This step was carried out according to the method described in <NPL>. A suspension of activated zinc (<NUM>, <NUM> mmol) in pyridine (<NUM>) were successively added methyl acrylate (<NUM>, <NUM> mmol) and NiCl<NUM>·<NUM><NUM>O (<NUM>, <NUM> mmol) at room temperature. The mixture was stirred at <NUM> for <NUM> and then cooled to <NUM>. A solution of iodide <NUM> (<NUM>, <NUM> mmol) in pyridine (<NUM>) was added dropwise. The reaction mixture was stirred for <NUM>, and then diluted with ethyl acetate. The mixture was filtered through a pad of celite. The filtrate was washed with <NUM> HCl aq. two times, and the aqueous layer was extracted with ethyl acetate three times. The combined organic extracts were washed with H<NUM>O and brine, dried over MgSO<NUM>, and concentrated in vacuo. The pyridine hydrochloride was removed by filtering through a plug of cotton, and the filtrate was evaporated. The residue was chromatographed on silica gel (hexane/ethyl acetate; <NUM>:<NUM> to <NUM>:<NUM>) to give methyl ester <NUM> (<NUM>, <NUM>%).

Activated magnesium (<NUM>, <NUM> mmol) in dry diethyl ether (<NUM>) was added methyl iodide (<NUM>, <NUM> mmol) dropwise at <NUM>. To a solution of methyl ester <NUM> (<NUM>, <NUM> mmol) in dry diethyl ether (<NUM>) was added above Grignard reagent (<NUM>, <NUM>) dropwise at -<NUM>. The mixture was stirred for <NUM> at same temperature and then warmed to room temperature. After stirring for <NUM>, the reaction was quenched by careful addition of sat. NH<NUM>Cl aq. The aqueous layer was extracted with ethyl acetate three times. The combined organic extracts were washed with brine, dried over MgSO<NUM>, and concentrated in vacuo. The residue was chromatographed on silica gel (Hexane/ethyl acetate; <NUM>:<NUM> to <NUM>:<NUM>) to give diol <NUM> (<NUM>, <NUM>%).

A solution of diol <NUM> (<NUM>, <NUM> mmol) in dry acetonitrile (<NUM>) was added molecular sieves <NUM> angstroms (<NUM>) and N-methyl morpholine N-oxide (<NUM>, <NUM> mmol) and stirred for <NUM>. Tetraisopropylammonium perruthenate (<NUM>, <NUM> mmol) was added, then the reaction mixture was warmed to room temperature. After stirring for <NUM>, the mixture was filtered through a pad of silica gel, and the filtrate was evaporated to give crude ketone. A suspension of (bromomethyl)triphenylphosphonium bromide (<NUM>, <NUM> mmol) in toluene (<NUM>) was sonicated for <NUM> at room temperature and evaporated with toluene (<NUM>) three times at <NUM>. To the suspension was added dry THF (<NUM>) and cooled to <NUM>. Sodium bis(trimethylsilyl)amide in THF (<NUM>, <NUM> mmol, <NUM>) was added dropwise. After stirring for <NUM>, a solution of above crude ketone (<NUM> mmol) in THF (<NUM>) was added dropwise. After additive <NUM>, the reaction was quenched by the addition of a few drops of sat. NH<NUM>Cl aq. The mixture was filtered, and the filtrate was evaporated. The residue was chromatographed on silica gel (hexane/ethyl acetate; <NUM>:<NUM> to <NUM>:<NUM>) to give bromo olefin <NUM> (<NUM>, <NUM>%, <NUM> steps).

A solution of bromo olefin <NUM> (<NUM>, <NUM> mmol) in CH<NUM>Cl<NUM> (<NUM>) was added <NUM>,<NUM>-lutidine (<NUM>, <NUM> mmol) and dropwised triethylsilyl trifluoromethanesulfonate (<NUM>, <NUM> mmol). After stirring for <NUM>, the reaction was quenched by sat. NaHCO<NUM>. The aqueous layer was extracted with CH<NUM>Cl<NUM> three times. The combined organic extracts were dried over MgSO<NUM>, and concentrated in vacuo. The residue was chromatographed on silica gel (hexane/dichloromethane; <NUM>:<NUM>) to give diol <NUM> (<NUM>, <NUM>%). <NUM>H NMR (<NUM>, CDCl<NUM>) δ <NUM> (s, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM> (s, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM> (s, <NUM>), <NUM> (q, J = <NUM>, <NUM>).

To a solution of <NUM>-[<NUM>-(trifluoromethyl)-<NUM>H-diazirin-<NUM>-yl]benzoic acid (<NUM>, <NUM> mmol; purchased from Tokyo Chemical Industry Co. ) and N-hydroxysuccinimide (<NUM>, <NUM> mmol) in dichloromethane (<NUM>) was added N,N'-dicyclohexylcarbodiimide (<NUM>, <NUM> mmol) at <NUM> and the mixture was stirred at room temperature overnight. The precipitation was filtered off and the filtrate was concentrated under reduced pressure to give a crude product. To a solution of <NUM>-methyl L-glutamate (<NUM>, <NUM> mmol) in acetonitrile/H<NUM>O (<NUM>:<NUM>, <NUM>) was added the crude product and triethylamine (<NUM>, <NUM> mmol) and the mixture was stirred at room temperature overnight. The mixture was evaporated and the residue was dissolved in ethyl acetate, washed with aq. HCl (<NUM>), water, brine, and dried (MgSO<NUM>). The solvent was evaporated to afford crude product <NUM> (<NUM>), which was used in next step without further purification.

To a mixture of crude <NUM> (<NUM>) and N-Boc-<NUM>,<NUM>,<NUM>-trioxa-<NUM>,<NUM>-tridecanediamine (<NUM>, <NUM>. 15mmol) in dichloromethane (<NUM>) was added <NUM>-(<NUM>-dimethylaminopropyl)-<NUM>-ethylcarbodiimide hydrochloride (<NUM>, <NUM> mmol), <NUM>-dimethylaminopyridine (<NUM>, <NUM> mmol) and <NUM>-hydroxybenzotriazole monohydrate (<NUM>, <NUM> mmol) at <NUM>, and the mixture was stirred at room temperature overnight. The mixture was washed with sat. NaHCO<NUM>, water, and brine. The organic layer was dried over anhydrous magnesium sulfate and concentrated in vacuo. The residue was purified by column chromatography (ethyl acetate:hexane; <NUM>:<NUM>) to yield <NUM> (<NUM>, <NUM> mmol, <NUM>% for <NUM> steps).

To a solution of <NUM> (<NUM>, <NUM> mmol) in dichloromethane (<NUM>) was added trifluoroacetic acid (<NUM>) at <NUM> and the mixture was stirred at room temperature for <NUM>. The solvent was evaporated and the residue was dissolved in chloroform, washed with sat. Na<NUM>CO<NUM>, water and brine. The organic layer was dried over anhydrous magnesium sulfate and concentrated in vacuo to afford a crude product (<NUM>). To a solution of the crude product (<NUM>) in N,N'-dimethylformamide (<NUM>) was added NHS-Biotin (<NUM>, <NUM> mmol) and triethylamine (<NUM>, <NUM> mmol), and the mixture was stirred at room temperature overnight. The solvent was evaporated and the residue was purified by column chromatography (methanol: chloroform; <NUM>:<NUM>) to yield <NUM> (<NUM>, <NUM> mmol, <NUM>% for steps).

To a mixture of <NUM> (<NUM>, <NUM> mmol) in tetrahydrofuran (<NUM>) and water (<NUM>) was added lithium hydroxide monohydrate (<NUM>, <NUM> mmol) at <NUM> and the mixture was stirred at room temperature for <NUM>. The mixture was acidified with aq. HCl (<NUM>) and extracted with chloroform three times. The combined extracts were dried over anhydrous magnesium sulfate and concentrated in vacuo. The residue was purified by column chromatography (methanol:chloroform; <NUM>:<NUM>) to yield <NUM> (<NUM>, <NUM> mmol, <NUM>%).

This step was carried out according to the method described in <NPL>. <NUM>,<NUM>-Diol <NUM> (<NUM>, <NUM> mmol) was obtained in <NUM>% yield from D-(-)-Quinic acid <NUM> (<NUM>, <NUM> mmol).

A solution of diol <NUM> (<NUM>, <NUM> mmol) in CH<NUM>Cl<NUM> (<NUM>) was added benzaldehyde dimethyl acetal and pyridinium p-toluenesulfonate at room temperature. After stirring for <NUM>, the reaction was quenched by sat. NaHCO<NUM> aq. The aqueous layer was extracted with CH<NUM>Cl<NUM> three times. The combined organic extracts were dried over MgSO<NUM>, and concentrated in vacuo. The residue was chromatographed on silica gel (Hexane/dichloromethane; <NUM>:<NUM>) to give benzylidene acetal (<NUM>) including residual reagent (benzaldehyde dimethyl acetal). To a solution of above benzylidene acetal (<NUM>) in THF (<NUM>) was added a solution of TBAF·<NUM><NUM>O (<NUM>, <NUM> in THF) at <NUM>. After stirring for <NUM>, the reaction mixture was quenched with sat. NH<NUM>Cl aq. The aqueous layer was extracted with ethyl acetate three times. The combined organic extracts were washed with brine, dried over MgSO<NUM>, and concentrated in vacuo. The residue was chromatographed on silica gel (Hexane/ethyl acetate; <NUM>:<NUM> to <NUM>:<NUM>) to give alcohol <NUM> (<NUM>, <NUM>%).

A solution of alcohol <NUM> (<NUM>, <NUM> mmol) in CH<NUM>Cl<NUM> (<NUM>) was added pyridine (<NUM>, <NUM> mmol) and chloromethylsulfonyl chloride (<NUM>, <NUM> mmol) at <NUM>. After stirring for <NUM>, the reaction was quenched with sat. NH<NUM>Cl aq. The aqueous layer was extracted with CH<NUM>Cl<NUM> three times. The combined organic extracts were dried over MgSO<NUM>, and concentrated in vacuo to give crude monochlate. To a solution of above crude monochlate in DMF (<NUM>) was added sodium azide (<NUM>, <NUM> mmol) at room temperature, then the mixture was warmed to <NUM> and stirred for <NUM>. The reaction mixture was quenched with H<NUM>O. The aqueous layer was extracted with ethyl ether three times. The combined organic extracts were washed with brine, dried over MgSO<NUM>, and concentrated in vacuo. The residue was chromatographed on silica gel (Hexane/ethyl acetate; <NUM>:<NUM>) to give azide <NUM> (<NUM>, <NUM>% from <NUM>).

A solution of trimethylphosphine (<NUM>, <NUM> mmol, <NUM> in toluene) and distilled water (<NUM>) was added to a solution of azide <NUM> (<NUM>, <NUM> mmol) in THF (<NUM>) at room temperature. After stirring for <NUM>, the mixture was concentrated at <NUM> to give crude amine. To a solution of above crude amine in THF (<NUM>) was added N-carbethoxy phthalimide (<NUM>, <NUM> mmol) at room temperature. After stirring for <NUM>, to the mixture was added sat. Na<NUM>CO<NUM> aq. (<NUM>) and stirred vigorously. After additive <NUM>, the aqueous layer was extracted with ethyl ether three times. The combined organic extracts were washed with brine, dried over MgSO<NUM>, and concentrated in vacuo. The residue was chromatographed on silica gel (Hexane/ethyl acetate; <NUM>:<NUM> to <NUM>:<NUM>) to give azide <NUM> (<NUM>, <NUM>% from <NUM>).

A solution of phthalimide <NUM> (<NUM>, <NUM> mmol) in ethanol (<NUM>) was added palladium hydroxide (<NUM>, <NUM> wt%, <NUM>% on Carbon wetted with ca. <NUM>% Water from TCI Co. ) at room temperature. The reaction vessel was purged and placed under hydrogen and stirred vigorously for <NUM>. The reaction mixture was filterd through a pad of celite and filtrate was concentrated. The residue was chromatographed on silica gel (Hexane/ethyl acetate; <NUM>:<NUM> to <NUM>:<NUM>) to give diol <NUM> (<NUM>, <NUM>%) and starting material (<NUM>, <NUM>% recovery).

To a solution of <NUM> (<NUM>, <NUM> mmol) in methanol (<NUM>) was added dropwise a solution of sodium periodate (<NUM>, <NUM> mmol) in distilled water (<NUM>) at <NUM>. After stirring for <NUM>, the reaction mixture was diluted with water and the aqueous layer was extracted with dichloromethane three times. The combined organic extracts were washed with brine, dried over MgSO<NUM>, and concentrated in vacuo. The residue was chromatographed on silica gel (Hexane/ethyl acetate; <NUM>:<NUM> to <NUM>:<NUM>) to give ketone <NUM> (<NUM>, <NUM>%) and starting material <NUM> (<NUM>, <NUM>% recovery). <NUM>H NMR (<NUM>, CDCl<NUM>) δ <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM> (dddd, J= <NUM>, <NUM>, <NUM>, <NUM>, <NUM>), <NUM> (dddd, J = <NUM>, <NUM>, <NUM>, <NUM>, <NUM>), <NUM> (dd, J = <NUM>, <NUM>, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>).

A solution of diol <NUM> (<NUM>, <NUM> mmol) in dry dichloromethane (<NUM>) was added molecular sieves 4A (<NUM>) and N-methyl morpholine A-oxide (<NUM>, <NUM> mmol) and stirred for <NUM>. Tetraisopropylammonium perruthenate (<NUM>, <NUM> mmol) was added, then the reaction mixture was warmed to room temperature. After stirring for <NUM>, the mixture was filtered through a pad of silica gel, and the filtrate was evaporated to give crude ketone. A suspension of sodium hydride (<NUM>, <NUM> mmol, <NUM>% stabilized by oil) in THF (<NUM>) was added triethyl phosphonoacetate (<NUM>, <NUM> mmol) at <NUM> and warmed to room temperature. To the solution was added a solution of above ketone (<NUM> mmol) in THF (<NUM>). After stirring for <NUM> days, the reaction was quenched with H<NUM>O and aqueous layer was extracted with ethyl acetate three times. The combined organic extracts were washed with brine, dried over MgSO<NUM>, and concentrated in vacuo. The residue was chromatographed on silica gel (Hexane/ethyl acetate; <NUM>:<NUM> to <NUM>:<NUM>) to give α,β-unsaturated ester <NUM> (<NUM>, <NUM>%) as a colorless oil. <NUM>H NMR (<NUM>, CDCl<NUM>) δ <NUM> (s, <NUM>), <NUM> (q, J = <NUM>, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM>(m, <NUM>), <NUM> (t, J= <NUM>, <NUM>), <NUM> (s, <NUM>), <NUM> (d, J= <NUM>, <NUM>), <NUM> (s, <NUM>).

A solution of α,β-unsaturated ester <NUM> (<NUM>, <NUM> mmol) in CH<NUM>Cl<NUM> (<NUM>) was added diisobutyl aluminum hydride in hexane (<NUM>, <NUM> mmol, <NUM>). After stirring for <NUM>, the reaction was quenched by careful addition of MeOH (<NUM>). The mixture was added sat. Rochelle salt aq. (<NUM>) and stirred for <NUM>. The organic layer was washed with sat. Rochelle salt aq. three times, and the aqueous layer was extracted with CHCl<NUM> three times. The combined mixture was dried over MgSO<NUM> and concentrated in vacuo. The residue was chromatographed on silica gel (Hexane/ethyl acetate; <NUM>:<NUM> to <NUM>:<NUM>) to give allyl alcohol <NUM> (<NUM>, q. ) as a colorless oil. <NUM>H NMR (<NUM>, CDCl<NUM>) δ <NUM> (dd, J= <NUM>, <NUM>, <NUM>), <NUM> (dd, J= <NUM>, <NUM>, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM>(m, <NUM>), <NUM> (s, <NUM>), <NUM> (d, J = <NUM>, <NUM>), <NUM> (s, <NUM>).

Triphenylphosphine (<NUM>, <NUM> mmol), <NUM>-mercapt benzothiazole (<NUM>, <NUM> mmol) were added to a solution of allyl alcohol <NUM> (<NUM>, <NUM> mmol) in dichloromethane (<NUM>) at <NUM>, then added dropwise diisopropyl azodicarboxylate (<NUM>µL, <NUM> mmol). After stirring for <NUM>, the reaction mixture was quenched with H<NUM>O. The aqueous layer was extracted with dichloromethane three times. The combined organic extracts were dried over MgSO<NUM>, and concentrated in vacuo. The residue was chromatographed on silica gel (hexane/ethyl acetate; <NUM>:<NUM> to <NUM>:<NUM>) to thioether (<NUM> mmol) including residual reagent. To a solution of thioether (<NUM> mmol) in ethanol (<NUM>) was added hydrogen peroxide (<NUM>, <NUM>% solution in water) and hexaammonium heptamolybdate tetrahydrate (<NUM>, <NUM> mmol) at room temperature. After stirring for <NUM>, the reaction was quenched with <NUM>% Na<NUM>S<NUM>O<NUM> aq. The aqueous layer was extracted with ethyl acetate three times. The combined organic extracts were washed with brine, dried over MgSO<NUM>, and concentrated in vacuo. The residue was chromatographed on silica gel (hexane/ethyl acetate; <NUM>:<NUM> to <NUM>:<NUM>) to give alcohol <NUM> (<NUM>, <NUM>%).

To a solution of alcohol <NUM> (<NUM>, <NUM> mmol) in N,N-dimethylformamide (<NUM>) was added imidazole (<NUM>, <NUM> mmol) and N,N-dimethylamino-<NUM>-pyridine (<NUM>, <NUM> mmol) at room temperature, then added dropwise chloro triethylsilane (<NUM>, <NUM> mmol). After stirring for <NUM>, the reaction mixture was quenched with H<NUM>O. The aqueous layer was extracted with ethyl ether three times. The combined organic extracts were washed with H<NUM>O and brine, dried over MgSO<NUM>, and concentrated in vacuo. The residue was chromatographed on silica gel (hexane/ethyl acetate; <NUM>:<NUM> to <NUM>:<NUM>) to give benzothiazolyl sulfone <NUM> (<NUM>, <NUM>%).

A solution of alcohol <NUM> (<NUM>, <NUM> mmol) in CH<NUM>Cl<NUM> (<NUM>) was added molecular sieves 4A and N-methylmorpholine N-oxide at room temperature. After stirring for <NUM>, tetrapropylammonium perruthenate was added at <NUM>. After additive <NUM>, the reaction mixture was filtered through a pad of silica gel and filtrate was concentrated. The residue was chromatographed on silica gel (Hexane/dichloromethane; <NUM>:<NUM> to <NUM>:<NUM>) to give ketone 78a (<NUM>, <NUM>%) and 78b (<NUM>, <NUM>%).

A solution of ketone 78a (<NUM>, <NUM> mmol) in ethanol (<NUM>) was added sodium borohydride (<NUM>, <NUM> mmol). After stirring for <NUM>, the reaction mixture was quenched with H<NUM>O and brine, then aqueous layer was extracted with ethyl acetate three times. The combined organic extracts were washed with brine, dried over MgSO<NUM>, and concentrated in vacuo. The residue was chromatographed on silica gel (hexane/ethyl acetate; <NUM>:<NUM> to <NUM>:<NUM>) to give cis-alcohol 79a (<NUM>, <NUM>%) and trans-alcohol 62a (<NUM>, <NUM>%) as a colorless oil.

By following the same procedure described for 79a, cis-alcohol 79b (<NUM>, <NUM>%) and trans-alcohol 62b (<NUM>, <NUM>%) was obtained from 78b (<NUM>, <NUM> mmol) as a colorless oil.

By following the same procedure described for <NUM>, phthalimide <NUM> (<NUM>, <NUM>%) was obtained from 79a and 79b (<NUM>, <NUM> mmol) as an amorphous solid like foam.

A solution of phthalimide <NUM> (<NUM>, <NUM> mmol) in ethanol (<NUM>) was added palladium hydroxide (<NUM>, <NUM> wt%, <NUM>% on Carbon wetted with ca. <NUM>% Water from TCI Co. ) at room temperature. The reaction vessel was purged and placed under hydrogen and stirred vigorously for <NUM>. The reaction mixture was filterd through a pad of celite and filtrate was concentrated. The residue was chromatographed on silica gel (Hexane/ethyl acetate; <NUM>:<NUM> to <NUM>:<NUM>) to give diol (<NUM>, <NUM>%) and starting material (<NUM>, <NUM>% recovery). To a solution of above diol (<NUM>, <NUM> mmol) in methanol (<NUM>) was added dropwise a solution of sodium periodate (<NUM>, <NUM> mmol) in distilled water (<NUM>) at <NUM>. After stirring for <NUM>, the reaction mixture was diluted with water and the aqueous layer was extracted with dichloromethane three times. The combined organic extracts were washed with brine, dried over MgSO<NUM>, and concentrated in vacuo. The residue was chromatographed on silica gel (Hexane/ethyl acetate; <NUM>:<NUM> to <NUM>:<NUM>) to give ketone <NUM> (<NUM>, <NUM>%) as a colorless solid. <NUM>H NMR (<NUM>, CDCl<NUM>) δ <NUM>-<NUM> (m, <NUM>), <NUM> (dddd, J = <NUM>, <NUM>, <NUM>, <NUM>, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM> (dd, J= <NUM>, <NUM>, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>).

A solution of alcohol 79a and 79b (<NUM>, <NUM> mmol) in CH<NUM>Cl<NUM> (<NUM>) was added triethylamine (<NUM>, <NUM> mmol), N,N-dimethyl-<NUM>-aminopyridine (<NUM>, <NUM> mmol) and benzoyl chloride (<NUM>, <NUM> mmol) at <NUM>, then the mixture was warmed to room temperature and stirred for <NUM>. The reaction was quenched with sat. NaHCO<NUM> aq. The aqueous layer was extracted with CH<NUM>Cl<NUM> three times. The combined organic extracts were dried over MgSO<NUM>, and concentrated in vacuo. The residue was chromatographed on silica gel (Hexane/ethyl acetate; <NUM>:<NUM>) to give <NUM> (<NUM>, <NUM>%) as a colorless solid.

A solution of benzoate ester <NUM> (<NUM>, <NUM> mmol) in ethyl acetate (<NUM>) was added palladium hydroxide (<NUM>, <NUM> wt%, <NUM>% on Carbon wetted with ca. <NUM>% Water from TCI Co. ) at room temperature. The reaction vessel was purged and placed under hydrogen and stirred vigorously for <NUM>. The reaction mixture was filterd through a pad of celite and filtrate was concentrated. The residue was chromatographed on silica gel (Hexane/ethyl acetate; <NUM>:<NUM> to <NUM>:<NUM>) to give diol (<NUM>, <NUM>%) as a colorless solid. To a solution of above diol (<NUM>, <NUM> mmol) in methanol (<NUM>) was added dropwise a solution of sodium periodate (<NUM>, <NUM> mmol) in distilled water (<NUM>) at <NUM>. After stirring for <NUM>, the reaction mixture was diluted with water and the aqueous layer was extracted with dichloromethane three times. The combined organic extracts were washed with brine, dried over MgSO<NUM>, and concentrated in vacuo. The residue was chromatographed on silica gel (Hexane/ethyl acetate; <NUM>:<NUM> to <NUM>:<NUM>) to give ketone <NUM> (<NUM>, <NUM>%) as a colorless solid.

To a solution of <NUM> (<NUM>, <NUM> mmol) and 14a (<NUM>, <NUM> mmol) in toluene (<NUM>) and Et<NUM>N (<NUM>) was added Pd(PPh<NUM>)<NUM> (<NUM>-<NUM> mol%) at room temperature, then the reaction mixture was heated at <NUM>. After stirring for <NUM>, the reaction mixture was filtered through a pad of silica gel column and the filtrates were evaporated. The residue was chromatographed on silica gel (hexane/ethyl acetate; <NUM>:<NUM>) to give 17a (<NUM>, <NUM> mmol, not pure). To a solution of 17a (<NUM>, <NUM> mmol) in THF (<NUM>) was added 3HF-Et<NUM>N (<NUM>, <NUM> mmol) at room temperature. The reaction was monitored by thin layer chromatography on silica gel plates (eluent: <NUM>:<NUM> ethyl acetate to hexane). The reaction was quenched by sat. NaHCO<NUM>, the aqueous layer was extracted with ethyl acetate three times. The combined organic extracts were washed with brine, dried over MgSO<NUM>, and concentrated in vacuo. The residue was chromatographed on silica gel (hexane/ethyl acetate; <NUM>:<NUM> to <NUM>:<NUM>) to give 1α-NHBoc VD<NUM> 18a (<NUM>, <NUM> steps <NUM>%).

By following the same procedure described for 18a, 1β-NHBoc VD<NUM> 18b (<NUM>, <NUM>%) was obtained from <NUM> (<NUM>, <NUM> mmol) and 14b (<NUM>, <NUM> mmol) as a colorless oil.

Acetyl chloride (<NUM>, <NUM> mmol) was slowly added to EtOH (<NUM>) dropwise at <NUM>. After stirring for <NUM>, this solution was added to diol 18a (<NUM>, <NUM> mmol). After additional stirring for <NUM>, the reaction mixture was evaporated. The residue was washed with Et<NUM>O, affording 19a (<NUM>, <NUM>%) as a yellow solid.

By following the same procedure described for 19a, 19b (<NUM>, <NUM>%) was obtained from 18b (<NUM>, <NUM> mmol) as a yellow solid. <NUM>H NMR (<NUM>, MeOH-d<NUM>) δ <NUM> (d, J = <NUM>, <NUM>), <NUM> (d, J= <NUM>, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM> (d, J = <NUM>, <NUM>), <NUM> (d,J = <NUM>, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM> (d, J= <NUM>, <NUM>), <NUM> (s, <NUM>).

To a solution of <NUM> (<NUM>, <NUM> mmol) and 20a (<NUM>, <NUM> mmol) in toluene (<NUM>) and Et<NUM>N (<NUM>) was added Pd(PPh<NUM>)<NUM> (<NUM> - <NUM> mol%) at room temperature, then the reaction mixture was heated at <NUM>. After stirring for <NUM>, the reaction mixture was filtered through a pad of silica gel column and the filtrates were evaporated. The residue was chromatographed on silica gel (hexane/ethyl acetate; <NUM>:<NUM> to <NUM>:<NUM>) to give 21a (<NUM>, <NUM>%). To a solution of 21a (<NUM>, <NUM> mmol) in THF (<NUM>) was added 3HF-Et<NUM>N (<NUM>, <NUM> mmol) at room temperature. The reaction was monitored by thin layer chromatography on silica gel plates (eluent: <NUM>:<NUM>:<NUM> chloroform to ethyl acetate to methanol). The reaction mixture was quenched with sat. NaHCO<NUM> aq. The aqueous layer was extracted with ethyl acetate three times. The combined organic extracts were washed with brine, dried over MgSO<NUM>, and concentrated in vacuo. The residue was chromatographed on silica gel (hexane/ethyl acetate; <NUM>:<NUM> to <NUM>:<NUM>) to give 1α-NHAc VD<NUM> 22a (<NUM>, <NUM>%).

By following the same procedure described for 22a, 1β-NHAc VD<NUM> 22b (<NUM>, <NUM>%) was obtained from <NUM> (<NUM>, <NUM> mmol) and 20b (<NUM>, <NUM> mmol) as colorless oil. <NUM>H NMR (<NUM>, CDCl<NUM>) δ <NUM> (d, J = <NUM>, <NUM>), <NUM> (d, J = <NUM>, <NUM>), <NUM> (d, J= <NUM>, <NUM>), <NUM> (s, <NUM>), <NUM> (s,<NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM> (d, J = <NUM>, <NUM>), <NUM> (d, J = <NUM>, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (d, J = <NUM>, <NUM>), <NUM> (s, <NUM>).

To a solution of <NUM> (<NUM>, <NUM> mmol) and <NUM> (<NUM>, <NUM> mmol) in toluene (<NUM>) and Et<NUM>N (<NUM>) was added Pd(PPh<NUM>)<NUM> (<NUM>-<NUM> mol%) at room temperature, then the reaction mixture was heated at <NUM>. After stirring for <NUM>, the reaction mixture was filtered through a pad of silica gel column and the filtrates were evaporated. The residue was chromatographed on silica gel (hexane/ethyl acetate; <NUM>:<NUM>) to give <NUM> (<NUM>, <NUM>%). To a solution of <NUM> (<NUM>, <NUM> mmol) in THF (<NUM>) was added 3HF-Et<NUM>N (<NUM>, <NUM> mmol) at room temperature. The reaction was monitored by thin layer chromatography on silica gel plates (eluent: <NUM>:<NUM> ethyl acetate to hexane). The reaction was quenched by sat. NaHCO<NUM>. The aqueous layer was extracted with ethyl acetate three times. The combined organic extracts were washed with brine, dried over MgSO<NUM>, and concentrated in vacuo. The residue was chromatographed on silica gel (hexane/ethyl acetate; <NUM>:<NUM> to <NUM>:<NUM>) to give 1α-NHNs VD<NUM> <NUM> (<NUM>, <NUM>%).

To a solution of <NUM> (<NUM>, <NUM> mmol) and <NUM> (<NUM>, <NUM> mmol) in toluene (<NUM>) and Et<NUM>N (<NUM>) was added Pd(PPh<NUM>)<NUM> (<NUM>-<NUM> mol%) at room temperature, then the reaction mixture was heated at <NUM>. After stirring for <NUM>, the reaction mixture was filtered through a pad of silica gel column and the filtrate was evaporated. The residue was chromatographed on silica gel (hexane/ethyl acetate; <NUM>:<NUM>) to give <NUM> (<NUM>, <NUM>%). To a solution of <NUM> (<NUM>, <NUM> mmol) in THF (<NUM>) was added 3HF-Et<NUM>N (<NUM>, <NUM> mmol) at room temperature. The reaction was monitored by thin layer chromatography on silica gel plates (eluent: <NUM>:<NUM> ethyl acetate to hexane). The reaction mixture was quenched with sat. NaHCO<NUM> aq. The aqueous layer was extracted with ethyl acetate three times. The combined organic extracts were washed with brine, dried over MgSO<NUM>, and concentrated in vacuo. The residue was chromatographed on silica gel (hexane/ethyl acetate; <NUM>:<NUM> to <NUM>:<NUM>) to give <NUM>α-NHTs VD<NUM> <NUM> (<NUM>, <NUM>%).

To a solution of <NUM> (<NUM>, <NUM> mmol) and <NUM> (<NUM>, <NUM> mmol) in toluene (<NUM>) and Et<NUM>N (<NUM>) was added Pd(PPh<NUM>)<NUM> (<NUM>-<NUM> mol%) at room temperature, then the reaction mixture was heated at <NUM>. After stirring for <NUM>, the reaction mixture was filtered through a pad of silica gel column and the filtrates were evaporated. The residue was chromatographed on silica gel (hexane/ethyl acetate; <NUM>:<NUM> to <NUM>:<NUM>) to give <NUM> (<NUM>, <NUM>%). To a solution of <NUM> (<NUM>, <NUM> mmol) in THF (<NUM>) was added 3HF-Et<NUM>N (<NUM>, <NUM> mmol) at room temperature. The reaction was monitored by thin layer chromatography on silica gel plates (eluent: <NUM>:<NUM> ethyl acetate to hexane). The reaction mixture was quenched with sat. NaHCO<NUM> aq. The aqueous layer was extracted with ethyl acetate three times. The combined organic extracts were washed with brine, dried over MgSO<NUM>, and concentrated in vacuo. The residue was chromatographed on silica gel (Hexane/ethyl acetate; <NUM>:<NUM> to <NUM>:<NUM>) to give <NUM>α-NHCbz VD<NUM> <NUM> (<NUM>, <NUM>%).

A solution of <NUM>-dodecanethiol (<NUM>, <NUM> mmol) in diethyl ether (<NUM>) was added NaH (<NUM>, <NUM> mmol, <NUM>% stabilized in mineral oil) at <NUM> and stirred for <NUM>. To the suspension was added nosylate <NUM> (<NUM>, <NUM> mmol) in diethyl ether (<NUM>), and then the reaction mixture was warmed to room temperature. After additive <NUM>, the reaction was quenched by H<NUM>O. The aqueous layer was extracted with ethyl acetate three times. The combined organic extracts were washed with brine, dried over MgSO<NUM>, and concentrated in vacuo. The residue was chromatographed on silica gel (hexane/ethyl acetate; <NUM>:<NUM> to <NUM>:<NUM>) to give amine <NUM> (<NUM>, <NUM>%). <NUM>H NMR (<NUM>, CDCl<NUM>) δ <NUM> (d, J = <NUM>, <NUM>), <NUM> (d, J= <NUM>, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM> (s, <NUM>), <NUM> (t, J= <NUM>, <NUM>), <NUM> (s, <NUM>), <NUM> (q, J = <NUM>, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>).

A solution of amine <NUM> (<NUM>, <NUM> mmol) in CH<NUM>Cl<NUM> (<NUM>) was added Et<NUM>N (<NUM>, <NUM> mmol), and carefully dropwised p-(trifluoromethyl)benzoyl chloride (<NUM>, <NUM> mmol) at - <NUM>. After stirring for <NUM>, the reaction was quenched by sat. NaHCO<NUM> aq. The aqueous layer was extracted with ethyl acetate three times. The combined organic extracts were washed with brine, dried over MgSO<NUM>, and concentrated in vacuo. The residue was chromatographed on silica gel (hexane/ethyl acetate; <NUM>:<NUM> to <NUM>:<NUM>) to give benzamide (<NUM>, <NUM>%, not pure). To a solution of the benzamide (<NUM>, <NUM>µmol) in THF (<NUM>) was added 3HF-Et<NUM>N (<NUM>, <NUM> mmol) at room temperature. The reaction was monitored by thin layer chromatography on silica gel plates (eluent: <NUM>:<NUM> ethyl acetate to hexane). The reaction mixture was quenched with sat. NaHCO<NUM>. The aqueous layer was extracted with ethyl acetate three times. The combined organic extracts were washed with brine, dried over MgSO<NUM>, and concentrated in vacuo. The residue was chromatographed on silica gel (hexane/ethyl acetate; <NUM>:<NUM> to <NUM>:a1) to give 1α-NH(p-CF<NUM>Bz) VD<NUM> <NUM> (<NUM>, <NUM>%).

By following the similar procedure described for <NUM>, 1α-NH(p-CF<NUM>Bz) VD<NUM> <NUM> (<NUM>, <NUM>%) purified by flash chromatography (Hexane/EtOAc; <NUM>:<NUM> to <NUM>:<NUM>) was obtained from <NUM> (<NUM>, <NUM>µmol) as a solid.

By following the similar procedure described for <NUM>, 1α-NH(p-OMeBz) VD<NUM> <NUM> (<NUM>, <NUM>%) purified by flash chromatography (hexane/ethyl acetate; <NUM>:<NUM> to <NUM>:<NUM>) was obtained from <NUM> (<NUM>, <NUM>µmol) as a solid.

By following the similar procedure described for <NUM>, 1α-NH(p-SCF<NUM>Bz) VD<NUM> <NUM> (<NUM>, <NUM>%) purified by flash chromatography (hexane/ethyl acetate; <NUM>:<NUM> to <NUM>:<NUM>) was obtained from <NUM> (<NUM>, <NUM> mmol) as a solid.

By following the similar procedure described for <NUM>, 1α-NH(<NUM>,<NUM>,<NUM>,<NUM>-tetrafluoroBz) VD<NUM> <NUM> (<NUM>, <NUM>%) purified by flash chromatography (NH silica gel, hexane/ethyl acetate; <NUM>:<NUM>) was obtained from <NUM> (<NUM>, <NUM>µmol) as a solid.

By following the similar procedure described for <NUM>, 1α-NH(<NUM>,<NUM>,<NUM>-trifluoroBz) VD<NUM> <NUM> (<NUM>, <NUM>%) purified by flash chromatography (NH silica gel, hexane/ethyl acetate; <NUM>:<NUM>) was obtained from <NUM> (<NUM>, <NUM> mmol) as a solid.

By following the similar procedure described for <NUM>, 1α-NH(<NUM>,<NUM>-dimethoxyBz) VD<NUM> <NUM> (<NUM>, <NUM>%) purified by flash chromatography (hexane/ethyl acetate; <NUM>:<NUM>) was obtained from <NUM> (<NUM>, <NUM>µmol) as a solid.

By following the similar procedure described for <NUM>, 1α-NH(propionyl) VD<NUM> <NUM> (<NUM>, <NUM>%) purified by flash chromatography (chloroform/ethyl acetate/methanol; <NUM>:<NUM>:<NUM>) was obtained from <NUM> (<NUM>, <NUM>µmol) as a solid.

By following the similar procedure described for <NUM>, 1α-NH(butyryl) VD<NUM> <NUM> (<NUM>, <NUM>%) purified by flash chromatography (chloroform/methanol; <NUM>:<NUM>) was obtained from <NUM> (<NUM>, <NUM>µmol) as a solid.

Triphenylphosphine (<NUM>, <NUM> mmol), ethanol (<NUM>µL, <NUM> mmol) and diisopropyl azodicarboxylate (<NUM>µL, <NUM> mmol) were added to a solution of nosylamide <NUM> (<NUM>, <NUM> mmol) in THF (<NUM>) at room temperature and stirred for <NUM>. The reaction mixture was quenched with H<NUM>O. The aqueous layer was extracted with ethyl acetate three times. The combined organic extracts were washed with brine, dried over MgSO<NUM>, and concentrated in vacuo. The residue was chromatographed on silica gel (hexane/ethyl acetate; <NUM>:<NUM>) to give ethylated nosylamide (<NUM>, <NUM>%, not pure). To a solution of <NUM>-dodecanethiol (<NUM>µL, <NUM> mmol) in diethyl ether (<NUM>µL) was added sodium hydride (<NUM>, <NUM> mmol, <NUM>% stabilized by oil) at <NUM> and stirred for <NUM>. The suspension was added to a solution of above ethylated nosylamide (<NUM>, <NUM> mmol) in diethyl ether (<NUM>µL). After additive <NUM>, the reaction was quenched with H<NUM>O. The aqueous layer was extracted with ethyl acetate three times. The combined organic extracts were washed with brine, dried over MgSO<NUM>, and concentrated in vacuo. The residue was chromatographed on silica gel (hexane/ethyl acetate; <NUM>:<NUM> to <NUM>:<NUM>) to give ethyl amine (<NUM>, <NUM>%). HF Py (<NUM>, <NUM> mmol) was added to a solution of ethyl amine (<NUM>, <NUM>µmol) in THF (<NUM>) at room temperature. The reaction was monitored by thin layer chromatography on silica gel plates (eluent: <NUM>:<NUM>:<NUM> chloroform to ethyl acetate to methanol). The reaction mixture was quenched with sat. NaHCO<NUM> aq. The aqueous layer was extracted with ethyl acetate three times. The combined organic extracts were washed with brine, dried over MgSO<NUM>, and concentrated in vacuo. The residue was chromatographed on silica gel (NH silica gel, chloroform/methanol; <NUM>:<NUM>) to give 1α-NHEt VD<NUM> <NUM> (<NUM>, <NUM>%).

By following the similar procedure described for <NUM>, 1α-NHBu VD<NUM> <NUM> (<NUM>, <NUM>%) purified by flash chromatography (chloroform/methanol; <NUM>:<NUM>) was obtained from <NUM> (<NUM>, <NUM> mmol) as a solid.

By following the similar procedure described for <NUM>, 1α-NH(cyclopropylmethyl) VD<NUM> <NUM> (<NUM>, <NUM>%) purified by flash chromatography (chloroform/ethyl acetate/methanol; <NUM>:<NUM>:<NUM> to <NUM>:<NUM>:<NUM>) was obtained from <NUM> (<NUM>, <NUM> mmol) as a solid.

By following the similar procedure described for <NUM>, 1α-NH(<NUM>-morpholinoethyl) VD<NUM> <NUM> (<NUM>, <NUM>%) purified by flash chromatography (chloroform/ethyl acetate/methanol; <NUM>:<NUM>:<NUM> to chloroform/methanol; <NUM>:<NUM>) was obtained from <NUM> (<NUM>, <NUM> mmol) as a solid.

By following the similar procedure described for <NUM>, 1α-NH(<NUM>-(<NUM>-hydroxy piperidyl)ethyl) VD<NUM> <NUM> (<NUM>, <NUM>%) purified by flash chromatography (NH silica gel, chloroform/ethyl acetate/methanol; <NUM>:<NUM>:<NUM>) was obtained from <NUM> (<NUM>, <NUM> mmol) as a solid.

By following the similar procedure described for <NUM>, 1α-NHMs VD<NUM> <NUM> (<NUM>, <NUM>%) purified by flash chromatography (chloroform/methanol; <NUM>:<NUM>) was obtained from <NUM> (<NUM>, <NUM>µmol) as a solid.

By following the similar procedure described for <NUM>, 1α-NH(p-fluoroBz) VD<NUM> <NUM> (<NUM>, <NUM>%) purified by flash chromatography (NH silica, hexane/ethyl acetate; <NUM>:<NUM>) was obtained from <NUM> (<NUM>, <NUM>µmol) as a solid.

To a stirred solution of <NUM> (<NUM>, <NUM> mmol) and 19a (<NUM>, <NUM> mmol) in methanol (<NUM>) was added <NUM>-(<NUM>,<NUM>-dimethoxy-<NUM>,<NUM>,<NUM>-triazin-<NUM>-yl)-<NUM>-methylmorpholinium chloride (<NUM>, <NUM> mmol) and N,N-diisopropylethylamine (<NUM>µL, <NUM> mmol) at <NUM> and the mixture was stirred at room temperature overnight. The solvent was evaporated and the residue was purified by HPLC to yield <NUM> (<NUM>, <NUM> mmol, <NUM>%).

To a solution of benzothiazolyl sulfone <NUM> (<NUM>, <NUM> mmol) in THF (<NUM>) was added dropwise lithium bis(trimethylsilyl)amide (<NUM>µL, <NUM> mmol, <NUM> in THF) at -<NUM>. After stirring for <NUM>, a solution of ketone <NUM> (<NUM>, <NUM> mmol) in THF (<NUM>) was added. The mixture was stirred for <NUM> at same temperature, then warmed up to room temperature and stirred for <NUM>. The reaction mixture was quenched with water and the aqueous layer was extracted with ethyl acetate three times. The combined organic extracts were washed with brine, dried over MgSO<NUM>, and concentrated in vacuo. The residue was chromatographed on silica gel (Hexane/ethyl acetate; <NUM>:<NUM>) to give <NUM> (<NUM>, <NUM>%), <NUM> (<NUM>, <NUM>%) and starting material <NUM> (<NUM>, <NUM>% recovery).

To a solution of <NUM> (<NUM>, <NUM> mmol) in ethanol (<NUM>) was added hydrazine hydrate (<NUM>µL, <NUM> mmol) at room temperature. The mixture was stirred for <NUM> at <NUM>. The reaction mixture was filtered through a plug of cotton and filtrate was concentrated to give crude <NUM>.

By following the same procedure described for <NUM>, crude <NUM> was obtained from <NUM> (<NUM>, <NUM> mmol).

To a solution of crude <NUM> (ca. <NUM> mmol) in CH<NUM>Cl<NUM> (<NUM>) was added trimethylamine (<NUM>µL, <NUM> mmol) andp-toluenesulfonyl chloride (<NUM>, <NUM> mmol) at <NUM>. After stirring for <NUM>, the reaction mixture was quenched with water and the aqueous layer was extracted with dichloromethane three times, dried over MgSO<NUM>, and concentrated in vacuo. The residue was chromatographed on silica gel (NH silica gel, hexane/ethyl acetate; <NUM>:<NUM> to <NUM>:<NUM>) to give 1β-NHTs-3β-OTBS VD<NUM> (<NUM>, <NUM>%) as a colorless oil. HF Py (<NUM>, <NUM> mmol) was added to a solution of above 1β-NHTs-3β-OTBS VD<NUM> (<NUM>, <NUM>µmol) in THF (<NUM>) and the mixture was stirred for <NUM>. The reaction mixture was quenched with sat. NaHCO<NUM> aq. and aqueous layer was extracted with ethyl acetate three times. The combined organic extracts were washed with brine, dried over MgSO<NUM>, and concentrated in vacuo. The residue was chromatographed on silica gel (Hexane/ethyl acetate; <NUM>:<NUM> to <NUM>:<NUM>) to give 1β-NHTs-3β-OH VD<NUM> <NUM> (<NUM>, <NUM>%) as a white solid.

By following the same procedure described for <NUM>, 1α-OTBS-3α-NHTs VD<NUM> <NUM> (<NUM>, <NUM>%) was obtained from <NUM> (ca. <NUM> mmol) as a white solid.

To a solution of crude <NUM> (ca. <NUM> mmol) in CH<NUM>Cl<NUM> (<NUM>) was added trimethylamine (<NUM>µL, <NUM> mmol) and (p-trifluoromethylthio)benzoyl chloride (<NUM>µL, <NUM> mmol) at <NUM>. After stirring for <NUM>, the reaction mixture was quenched with water and the aqueous layer was extracted with dichloromethane three times, dried over MgSO<NUM>, and concentrated in vacuo. The residue was chromatographed on silica gel (hexane/ethyl acetate; <NUM>:<NUM> to <NUM>:<NUM>) to give 1β-NH(p-SCF<NUM>Bz)-3β-OTBS VD<NUM> (<NUM>, <NUM>%) as a colorless oil. HF Py (<NUM>, <NUM> mmol) was added to a solution of above 1β-NH(p-SCF<NUM>Bz)-3β-OTBS VD<NUM> (<NUM>, <NUM>µmol) in THF (<NUM>) and the mixture was stirred for <NUM>. The reaction mixture was quenched with sat. NaHCO<NUM> aq. and aqueous layer was extracted with ethyl acetate three times. The combined organic extracts were washed with brine, dried over MgSO<NUM>, and concentrated in vacuo. The residue was chromatographed on silica gel (Hexane/ethyl acetate; <NUM>:<NUM> to <NUM>:<NUM>) to give 1β-NH(p-SCF<NUM>Bz)-3β-OH VD<NUM> <NUM> (<NUM>, <NUM>%) as a white solid.

HF Py (<NUM>, <NUM> mmol) was added to a solution of above 1β-NPhth-3β-OTBS VD<NUM> <NUM> (<NUM>, <NUM>µmol) in THF (<NUM>) and the mixture was stirred for <NUM>. The reaction mixture was quenched with sat. NaHCO<NUM> aq. and aqueous layer was extracted with ethyl acetate three times. The combined organic extracts were washed with brine, dried over MgSO<NUM>, and concentrated in vacuo. The residue was chromatographed on silica gel (Hexane/ethyl acetate; <NUM>:<NUM> to <NUM>:<NUM>) to give 1β-NPhth-3β-OH VD<NUM> <NUM> (<NUM>, <NUM>%) as a white solid.

By following the same procedure described for <NUM>, 1α-OH-3α-NHPhth VD<NUM> <NUM> (<NUM>, <NUM>%) was obtained from <NUM> (<NUM>, <NUM>µmol) as a white solid.

To a solution of benzothiazolyl sulfone <NUM> (<NUM>, <NUM> mmol) in THF (<NUM>) was added dropwise lithium bis(trimethylsilyl)amide (<NUM>, <NUM> mmol, <NUM> in THF) at - <NUM>. After stirring for <NUM>, a solution of ketone <NUM> (<NUM>, <NUM> mmol) in THF (<NUM>) was added. The mixture was stirred for <NUM> at same temperature, then warmed up to room temperature and stirred for <NUM>. The reaction mixture was quenched with water and the aqueous layer was extracted with ethyl acetate three times. The combined organic extracts were washed with brine, dried over MgSO<NUM>, and concentrated in vacuo. The residue was chromatographed on silica gel (Hexane/ethyl acetate; <NUM>:<NUM> to <NUM>:<NUM>) to give <NUM> (<NUM>, <NUM>%), <NUM> (<NUM>, <NUM>%) and starting material <NUM> (<NUM>, <NUM>% recovery). <NUM>; <NUM>H NMR (<NUM>, CDCl<NUM>) δ <NUM>-<NUM> (m, <NUM>), <NUM> (d, J= <NUM>, <NUM>), <NUM> (d, J = <NUM>, <NUM>), <NUM> (dddd, J = <NUM>, <NUM>, <NUM>, <NUM>, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM> (s, <NUM>), <NUM>(t, J = <NUM>, <NUM>), <NUM> (d, J= <NUM>, <NUM>), <NUM> (s, <NUM>), <NUM> (q, J= <NUM>, <NUM>), <NUM> (s, <NUM>), <NUM> (s,<NUM>), <NUM> (s, <NUM>).

By following the same procedure described for <NUM>, crude <NUM> was obtained from <NUM> (<NUM>, <NUM>µmol).

By following the same procedure described for <NUM>, 1α-NHTs-3β-OH VD<NUM> <NUM> (<NUM>, <NUM>%) was obtained from crude <NUM> (ca. <NUM>µmol) as a white solid.

By following the same procedure described for <NUM>, 1α-NH(p-SCF<NUM>Bz)-3β-OH VD<NUM> <NUM> (<NUM>, <NUM>%) was obtained from crude <NUM> (ca. <NUM>µmol) as a white solid.

By following the same procedure described for <NUM>, 1α-NPhth-3β-OH VD<NUM> <NUM> (<NUM>, <NUM>%) was obtained from crude <NUM> (ca. <NUM>µmol) as a white solid.

By following the similar procedure described for <NUM>, 1α-NHAc-3β-OH VD<NUM> <NUM> (<NUM>, <NUM>%) was obtained from crude <NUM> (ca. <NUM>µmol) as a white solid.

By following the similar procedure described for <NUM>, 1α-NHMs-3β-OH VD<NUM> <NUM> (<NUM>, <NUM>%) was obtained from crude <NUM> (ca. <NUM>µmol) as a white solid.

By following the similar procedure described for <NUM>, 1α-NH(trifluoroacetyl)-3β-OH VD<NUM> <NUM> (<NUM>, <NUM>%) was obtained from crude <NUM> (ca. <NUM>µmol) as a white solid.

By following the similar procedure described for <NUM>, 1α-NHDns-3β-OH VD<NUM> <NUM> (<NUM>, <NUM>%) was obtained from crude <NUM> (ca. <NUM>µmol) as a white solid.

To a solution of benzothiazolyl sulfone <NUM> (<NUM>, <NUM> mmol) and ketone <NUM> (<NUM>, <NUM> mmol) in THF (<NUM>) was added dropwise lithium bis(trimethylsilyl)amide (<NUM>, <NUM> mmol, <NUM> in THF) at -<NUM>. The mixture was stirred for <NUM> at same temperature, then warmed up to room temperature and stirred for <NUM>. The reaction mixture was quenched with water and the aqueous layer was extracted with ethyl acetate three times. The combined organic extracts were washed with brine, dried over MgSO<NUM>, and concentrated in vacuo. The residue was chromatographed on silica gel (Hexane/ethyl acetate; <NUM>:<NUM> to <NUM>:<NUM>) to give <NUM> (<NUM>, <NUM>%) as isomer mixture and starting material <NUM> (<NUM>, <NUM>% recovery).

<NUM> (<NUM>, <NUM> mmol) was dissolved in methanol (<NUM>), then the solution was added potassium carbonate (<NUM>, <NUM> mmol) at room temperature and stirred for <NUM>. The reaction mixture was quenched with sat. NH<NUM>Cl aq. and the aqueous layer was extracted with dichloromethane three times. The combined organic extracts were dried over MgSO<NUM>, and concentrated in vacuo. The residue was chromatographed on silica gel (Hexane/ethyl acetate; <NUM>:<NUM> to <NUM>:<NUM>) to give <NUM> (<NUM>, <NUM>%) and isomer <NUM> (<NUM>, <NUM>%).

To a solution of <NUM> (<NUM>, <NUM> mmol) in dichloromethane (<NUM>) was added N,N-diethylaminosulfur trifluoride (<NUM>µL, <NUM> mmol) at -<NUM>. After stirring for <NUM>, the reaction mixture was quenched with sat. NaHCO<NUM> aq. and the aqueous layer was extracted with ethyl acetate three times. The combined organic extracts were dried over MgSO<NUM>, and concentrated in vacuo. The residue was chromatographed on silica gel (Hexane/ethyl acetate; <NUM>:<NUM> to <NUM>:<NUM>) to give <NUM> (<NUM>, <NUM>%).

HF Py (<NUM>, <NUM> mmol) was added to a solution of <NUM> (<NUM>, <NUM> mmol) in THF (<NUM>) and the mixture was stirred for <NUM>. The reaction mixture was quenched with sat. NaHCO<NUM> aq. and aqueous layer was extracted with ethyl acetate three times. The combined organic extracts were washed with brine, dried over MgSO<NUM>, and concentrated in vacuo. The residue was chromatographed on silica gel (Hexane/ethyl acetate; <NUM>:<NUM> to <NUM>:<NUM>) to give 1α-F-3β-OH VD<NUM> <NUM> (<NUM>, <NUM>%) as a white solid.

The molecules in a chemical library which consists of <NUM> endogenous lipid related molecules were screened for their ability to inhibit the expression of a luciferase reporter gene. In this gene, three repeats of sterol regulatory elements (SREs) control expression of luciferase. Lipid depletion activates SREBPs, which bind to the SRE domain and work as transcription factors to express luciferase. The SREBP-responsive reporter construct was co-transfected into Chinese hamster ovary K1 (CHO-K1) cells, along with a control β-galactosidase (β-gal) reporter gene. A constitutively active actin promoter in the reporter gene drives the expression of β-gal. The levels of luciferase expression from the SREBP-responsive reporter gene were normalized to the levels of β-gal expression.

Eleven molecules that decreased the expression levels of the reporter gene by more than <NUM>% were selected as hit molecules from the first screening. A reporter assay developed by <NPL>) was used to determine whether the selected hit molecules affected the ER-Golgi translocation and proteolytic processing of SREBPs, and to eliminate molecules that gave false positive results. In this assay, a secreted alkaline phosphatase, fused with an SREBP-<NUM> fragment lacking the NH<NUM>-terminal DNA-binding domain (PLAP-BP2<NUM>-<NUM>), was used to monitor translocation and processing, through changes in the fluorescence of a fluorogenic phosphatase substrate. When cells were co-transfected with plasmids encoding PLAP-BP2<NUM>-<NUM> and SREBP cleavage-activating protein (SCAP), PLAP phosphatase was secreted, and fluorescence signals were generated. Of the eleven hit molecules, vitamin D<NUM> (VD) derivatives caused a significant decrease in secretion, compared to the DMSO control (<FIG>). Two molecules in particular, <NUM>-hydroxyvitamin D<NUM> [<NUM>(OH)D] and 1α,<NUM>-dihydroxyvitamin D<NUM> [<NUM>,<NUM>(OH)<NUM>D], caused a greater decrease in PLAP secretion than did cholesterol.

CHO-K1 cells were maintained in a medium A (<NUM>:<NUM> mixture of Ham's F-<NUM> medium and Dulbecco's modified Eagle medium, supplemented with <NUM> units/mL penicillin, <NUM>µg/mL streptomycin sulfate, and <NUM>% (v/v) fetal bovine serum) at <NUM> under humidified <NUM>% CO<NUM>.

On day <NUM>, CHO-K1 cells were added to <NUM>-well plates at <NUM> × <NUM><NUM> cells per well in medium A. On day <NUM>, the cells were co-transfected with pCMV-PLAP-BP2<NUM>-<NUM> (<NUM>µg/well), pCMV-SCAP (<NUM>-<NUM>µg/well), and β-gal reporter, in which the expression of <NUM>-galactosidase was controlled by an actin promoter (pAc-β-gal, <NUM>µg/well) using FuGENE(Registered trademark) HD transfection reagent (Promega), according to the manufacturer's protocol. After <NUM> of incubation, the cells were washed with PBS and then incubated in medium B (<NUM>:<NUM> mixture of Ham's F-<NUM> medium and Dulbecco's modified Eagle medium, supplemented with <NUM> units/mL penicillin, <NUM>µg/mL streptomycin sulfate, <NUM>% (v/v) lipid-depleted serum, <NUM> compactin, and <NUM> lithium mevalonate), in the absence or presence of VD derivatives (<NUM>) or sterols (<NUM>µg/mL of cholesterol and <NUM>µg/mL of <NUM>-hydroxycholesterol). After <NUM> of incubation, an aliquot of the medium was assayed for secreted alkaline phosphatase activity. The cells in each well were lysed and used for measurement of β-galactosidase activities. The alkaline phosphatase activity was normalized by the activity of β-galactosidase.

<NUM>(OH)D inhibited the activation of SREBPs in a dose-dependent manner in the reporter assay, and had an IC<NUM> value of <NUM> (<FIG>, Table <NUM>). The inhibition of SREBP activation, mediated by the VD derivatives, was confirmed by Western blot analysis of SREBP-<NUM> (<FIG>). When cells were treated with <NUM>-hydroxycholesterol [<NUM>-HC], the mature form of SREBP disappeared, and the precursor form of SREBP accumulated, indicating that <NUM>-HC blocked the transportation of SREBP-SCAP complex from the ER to the Golgi apparatus. In contrast, treatment with <NUM>(OH)D decreased levels of both the mature and precursor forms, suggesting that the VD derivatives reduced the levels of the precursor form of SREBP, which consequently decreased levels of the mature form, resulting in the inhibition of the SREBP activation.

On day <NUM>, CHO-K1 cells were added to <NUM>-well plates at <NUM> × <NUM><NUM> cells per well in medium A. On day <NUM>, the cells were co-transfected with an SRE-<NUM>-driven luciferase reporter construct (pSRE-Luc) and pAc-β-gal at a <NUM>/<NUM> ratio, using FuGENE(Registered trademark) HD transfection reagent (Promega), according to the manufacturer's protocol. After <NUM>-<NUM> of incubation, the cells were washed with PBS buffer, then incubated in medium B containing the specific test compounds. Stock solutions of each compound in DMSO were prepared and added to medium B to give a <NUM>-fold (v/v) dilution (<NUM>% DMSO). After <NUM>-<NUM> of incubation, the cells in each well were lysed by freeze-thaw with Reporter Lysis Buffer (Promega), and aliquots were used to measure luciferase and β-galactosidase activities. Luciferase activity was measured using the Steady-Glo(Registered trademark) Luciferase Assay System (Promega), and β-galactosidase activity was measured using the β-galactosidase Enzyme Assay System (Promega). Luciferase activity was normalized to β-galactosidase activity.

On day <NUM>, CHO-K1 cells were added to <NUM>-well plates at <NUM> × <NUM><NUM> cells per well in medium A. On day <NUM>, the cells were washed with PBS, and then incubated in medium B in the absence or presence of specific test compounds (<NUM>). After <NUM> of incubation, the cells were washed three times with cold PBS, and lysed with a buffer A (<NUM> Tris-HCl (pH <NUM>), <NUM> NaCl, <NUM>% (v/v) Nonidet P40, <NUM>% (w/v) sodium deoxycholate, <NUM> urea, and protease inhibitor cocktail (Nacalai)). The cell lysate were passed <NUM> times through a <NUM> needle and centrifuged at <NUM>,<NUM> at <NUM> for <NUM>. The supernatants were transferred to new tubes, and the pellets were extracted with a buffer A. The resulting buffer was centrifuged at <NUM>,<NUM> at <NUM> for <NUM>, and the supernatants were combined to the previous ones. The resulting lysate was mixed with <NUM> volume of <NUM>× SDS sample buffer (Nacalai) and incubated at room temperature for <NUM>. The samples were separated on a <NUM>% SDS-PAGE gel and blotted, using mouse monoclonal antibodies against SREBP-<NUM> (IgG-7D4), SCAP (IgG-9D5), c-Myc (IgG1-MC045), and actin (AC-<NUM>). The specific bands were visualized using enhanced chemiluminescence (ECL Prime Western Blotting Detection Reagent, GE Healthcare) on an ImageQuant LAS <NUM> (GE Healthcare).

The effect of <NUM>(OH)D on the levels of SCAP was also confirmed by Western blot analysis (<FIG>). Treatment of cells with <NUM>(OH)D caused reduced the levels of SCAP as well as that of SREBP. Furthermore, the decrease in the levels of SCAP, mediated by <NUM>(OH)D, was cancelled by addition of MG-<NUM>, a general proteasome inhibitor (<FIG>), and <NUM>(OH)D accelerated the ubiquitination of SCAP (<FIG>). When SREBP is not able to form a complex with SCAP, the precursor form of SREBP is rapidly degraded (<NPL>. These results suggested that <NUM>(OH)D stimulates the ubiquitin-proteasome degradation of SCAP, resulting in the decrease in the levels of the precursor form of SREBP.

Despite the discovery of the SREBP inhibitory mechanism of hydroxylated VD derivatives, these endogenous molecules per se cannot be used for specific pharmacological intervention of SREBP due to their well-established roles in calcium homeostasis. For example, overdose administration <NUM>(OH)D, which is converted to an active VDR agonist, <NUM>,<NUM>(OH)<NUM>D, increases serum calcium concentrations, often resulting in formation of kidney stones. One of possible ways to eliminate its VDR activity is to prevent metabolic hydroxylation of the C1 position. Example compounds 18a, 22a, 22b, <NUM>, <NUM>, <NUM>, <NUM>, <NUM> and <NUM> are unlikely to be VDR agonists or converted to VDR agonists.

Reporter assays showed that these Example compounds inhibited activation of SREBPs, with IC<NUM> values ranging from <NUM> to <NUM> (Table <NUM>). The inhibition of SREBP activation mediated by Example compounds 22a, 22b, <NUM>, <NUM>, <NUM>, <NUM>, and <NUM> (<FIG>) and Example compounds <NUM>-<NUM>, <NUM>, <NUM>, <NUM>-<NUM>, <NUM>-<NUM>, and <NUM> (<FIG>) was confirmed by Western-blot analysis of SREBP-<NUM>. The treatment of cells with the derivatives decreased the levels of the mature form of SREBP relative to treatment with DMSO.

Claim 1:
A compound of the following general formula (I):
<CHM>
wherein one of RA and RB is hydroxyl and the other is NR<NUM>R<NUM>;
R<NUM> and R<NUM> are each independently selected from hydrogen; C<NUM>-<NUM> alkyl; C<NUM>-<NUM> alkylcarbonyl optionally substituted with at least one halogen which are the same or different; C<NUM>-<NUM> alkylsulfonyl; benzyloxycarbonyl; <NUM> to <NUM>-membered cycloalkyl-C<NUM>-<NUM> alkyl; C<NUM>-<NUM> arylcarbonyl optionally substituted with at least one group independently selected from the group consisting of halogen, halo-C<NUM>-<NUM> alkyl, -S-halo-C<NUM>-<NUM> alkyl, C<NUM>-<NUM> alkoxy, halo-C<NUM>-<NUM> alkoxy, nitro, cyano, C<NUM>-<NUM> alkoxycarbonyl and C<NUM>-<NUM> aryl; C<NUM>-<NUM> arylsulfonyl optionally substituted with at least one group independently selected from the group consisting of C<NUM>-<NUM> alkyl, nitro, and di-(C<NUM>-<NUM> alkyl)amino; <NUM> to <NUM>-membered saturated heterocyclyl-C<NUM>-<NUM> alkyl optionally substituted with at least one group independently selected from the group consisting of halogen and hydroxyl; <NUM> to <NUM>-membered heteroaryl; and a group of the following formula:
<CHM>
or
R<NUM> and R<NUM> may optionally combine together with the nitrogen atom to which they attach to form a nitrogen-containing oxo-substituted saturated <NUM> to <NUM>-membered heterocyclic ring which may be optionally fused with a C<NUM>-<NUM> aryl ring; and
R<NUM> is hydrogen or =CH<NUM>; or a pharmaceutically acceptable salt thereof;
provided that R<NUM> and R<NUM> are not concurrently hydrogen.