JP2001515064A - Synthesis of clast-lactacystin β-lactone and analogs thereof - Google Patents

Abstract

  (57) [Summary] The present invention is directed to an improved synthesis of crust-lactacystin-β-lactone, and its analogs, which proceeds with fewer steps and in a larger overall yield than the synthesis described in the prior art. The synthetic route relies on a novel configuration-specific synthesis of an oxazoline intermediate and a unique configuration-specific addition of formylamide to the oxazoline. Also described are novel crust-lactacystin-β-lactones, and analogs thereof, and their use as proteasome inhibitors.

Description

DETAILED DESCRIPTION OF THE INVENTION

FIELD OF THE INVENTION [0001] The present invention relates generally to methods for preparing lactacystin and related compounds, novel analogs of lactacystin and crust-lactacystin β-lactone, and their use as proteasome inhibitors.

2. Description of the Related Art Lactacystin (1), a metabolite of Streptomyces, inhibits cell cycle progression and induces neurite outgrowth in cultured neuroblastomas (Omura
J. et al. Antibiotics 44: 117 (1991); Omura
J. et al. Antibiotics 44: 113 (1991); Fentany et al., Pc. Natl. Acad. Sci. USA 91: 3358 (
1994)). The cellular target that mediates these effects is the proteolytic core of the 20S proteasome, the 26S proteasome, which is a central element of the ubiquitin-proteasome pathway of intracellular proteolysis. By mechanism research,
It has been established that lactacystin inhibits the proteasome through an intermediate step of the active species, a crust-lactacystin β-lactone (2) that specifically acylates the N-terminal threonine residue of the proteasome X / MB1 subunit. Yes (
Science 268: 726 (1995) by Fentany et al .; Di.
cck et al. Biol. Chem. 271: 7273 (1996)). Lactacystin analogs have been disclosed by Fentany et al. (WO
96/32105).

[0003]

Embedded image

[0004] The ubiquitin-proteasome pathway involves a variety of important physiological processes (Chemistry & Biology 2 by Goldberg et al.).
: 503 (1995); Cell 79:13 by Ciechanover (
1994); Trends Cell Biol. 5:
431 (1995)). In fact, cytoplasmic proteins are hydrolyzed by this pathway. First, protein substrates are marked for degradation by covalent attachment to multiple small protein molecules, ubiquitin. The resulting polyubiquitinated protein is then recognized and degraded by the 26S proteasome. Although its role in the degradation of damaged or mutated intracellular proteins has long been recognized, it is now known that this pathway also contributes to the selective degradation of various regulatory proteins. For example, ordered cell cycle progression requires programmed ubiquitination and degradation of cyclins. The ubiquitin-proteasome pathway also mediates the degradation of many other cell cycle regulatory proteins and tumor suppressor proteins (eg, p21, p27, p53).
Activation of the transcription factor NF-κB, which plays a central role in the regulation of genes involved in the immune and inflammatory responses, is dependent on ubiquitination and degradation of the inhibitory protein IκB-α (WO 95 / by Palombella et al. 2553
3). In addition, continuous metabolism of cellular proteins by the ubiquitin-proteasome pathway is essential for processing of antigenic peptides for presentation on MHC class I molecules (WO 94/17816 by Goldberg and Rock).
). The interesting biological activities of lactacystin and crust-lactacystin β-lactone, and the lack of natural materials, and the challenge of the chemical structure of the molecule, have stimulated synthetic efforts directed towards lactacystin and related analogs.
J. Corey and Reichard. Am. Chem. Soc. 114
: 10677 (1992); Tetrahedron Lett. 34: 697
7 (1993) achieved the first total synthesis of lactacystin proceeding in 15 steps and 10% overall yield. A key feature of the synthesis is the configuration-selective aldol reaction of cis-oxazolidinaldehyde from N-benzylserine to create C (6) -C (7) bonds. J. Uno et al. Am. Che
m. Soc. 116: 2139 (1994), the configuration-selective Mukaiyama-aldol reaction of a bicyclic oxazolidinyl silyl enol ether intermediate derived from D-pyroglutamic acid is converted to C (5).
-C (9) Used in fabrication. This synthesis comprises 19 steps and 5%
Proceeds in overall yield. Aldol reactions of similar bicyclic oxazolidine intermediates under basic conditions are described by Dikshit et al. In Tetrahedron Lett. Three
6: 6131 (1995).

The aldol reaction of oxazoline-derived enolates has been described by Smith and coworkers (J. Am. Chem. Soc. 115: 5302 by Suazuka et al.).
(1993); Nagamitsu et al. Am. Chem. Soc. 11
8: 3584 (1996)) and the synthesis of lactacystin, and Tetrahedron Lett. By Corey and Choi. 34
: 6969 (1993); Choi Ph. D. Thesis, Harvard University, 44 (1995), has a distinctive feature in the synthesis of (6R) -lactacystin. In the former synthesis, which proceeds in 20 steps and an overall yield of 9%, the enolate is condensed with formaldehyde to set up a single carbon atom, which has to be followed by a number of further steps. In the synthesis of Corey and Choi, the aldol reaction selectively yields the undesired stereochemical product resulting from the final preparation of the C (6) epimer of lactacystin, which lacks biological activity.

[0006] Lactacystin has also been prepared from D-glucose in 22 steps and a 2% overall yield (see J. Chem. Soc. Chem. Comm by Chida et al.).
un. 793 (1995)). The biosynthetic pathway involved in the production of natural products is ¹³ C
-Has been studied in feeding experiments involving enriched compounds (Takahedron Lett. 35: 5009 (1994) by Nakagawa et al.).

[0007] The reported synthesis of lactacystin is time consuming and proceeds in low yield.
Moreover, none of these syntheses are readily applied to analog synthesis. Thus, there is a need for improved methods of preparing lactacystin, crust-lactacystin β-lactone, and analogs thereof.

SUMMARY OF THE INVENTION [0008] A first aspect of the present invention relates to a process for forming lactacystin or an analog thereof having formula VI below, or clast-lactacystin β-lactone or an analog thereof having formula VII below:

Embedded image Wherein R ¹ is alkyl, alkenyl, alkynyl, cycloalkyl, aryl, alkaryl, or aralkyl, wherein any ring portion of the aryl, aralkyl, or alkaryl is optionally substituted. R ² is alkyl, cycloalkyl, aryl, alkaryl, aralkyl, alkoxy, hydroxy, alkoxyalkyl or amide, wherein any ring portion of said aryl, aralkyl or alkaryl is optionally substituted; R ⁷ is alkyl, aryl, aralkyl, or alkaryl (wherein any of the alkyl, aryl, aralkyl, or alkaryl may be optionally substituted)].

A second aspect of the present invention provides a compound of formula XIV:

Embedded image Wherein R ² is alkyl, cycloalkyl, aryl, alkaryl, aralkyl, alkoxy, hydroxy, alkoxyalkyl, or amide (wherein any ring moiety of the aryl, aralkyl or alkaryl is optionally substituted R ⁵ and R ⁶ are independently one of alkyl or alkaryl; alternatively, R ⁵ and R ⁶ are taken together with the nitrogen atom to which they are attached If
To form a 5- to 7-membered heterocyclic ring which may be optionally substituted and optionally further contain an oxygen or nitrogen atom. Is done.

A third aspect of the present invention provides a compound of formula Ia or Ib:

Embedded image Wherein R ¹ is alkyl, alkenyl, alkynyl, cycloalkyl, aryl,
R ⁴ is alkaryl or aralkyl, wherein any of the aryl, aralkyl or alkaryl ring moieties is optionally substituted; R ⁴ is aryl or heteroaryl, any of which is optionally Which may be substituted)]. The trisubstituted oxazolines of formulas Ia and Ib are useful as starting materials to form lactacystin, crust-lactacystin β-lactone, or analogs thereof, via the processes described herein.

A fourth aspect of the present invention is directed to lactacystin, crust-lactacystin β-lactone or analogs of formulas VI and VII having unexpected biological activity. Lactacystin, crust-lactacystin β-lactone, and their analogs have biological activity as inhibitors of the proteasome. They can be used to treat conditions directly mediated by proteasome function, such as muscle wasting, or indirectly through proteins processed by the proteasome, such as the transcription factor NF-κB. it can.

A fifth aspect of the present invention relates to a pharmaceutical composition comprising a compound of formula VI or VII and a pharmaceutically acceptable carrier or diluent.

[0013] A sixth aspect of the present invention relates to a method of inhibiting a proteasome function by administering a compound represented by the formula VI or the formula VII having a surprisingly high activity in inhibiting the proteasome. It relates to a method of treating a condition mediated directly or indirectly. A preferred embodiment is to use a compound of formula VI or VII to prevent infarction after vasculature or to reduce its size, for example to treat post-stroke neuronal loss. Be oriented. A further preferred embodiment is directed to the use of the compound for treating asthma.

[0014] A seventh aspect of the present invention relates to an enantiomerically-enriched composition of formylamide of formula XIV.

An eighth aspect of the present invention relates to an aldol represented by the formula II and a formula III
Novel individual intermediates such as aminodiols represented by:

Embedded image As well as individual steps within a multi-step process for forming lactacystin, crust-lactacystin β-lactone or various analogs thereof.

A ninth aspect of the present invention is directed to individual intermediates such as the compounds of formulas XVII, XVIII and XIX below:

Embedded image Wherein X is a halogen, preferably Cl, Br or I, as well as the individual steps in the multi-step process of forming the substituted oxazolines of the formula I.

[0017] Other features or advantages of the invention will be apparent from the following detailed description, and from the appended claims.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS The present invention relates to lactacystin, crust-lactacystin β-lactone and their processes, which proceed in fewer steps and with much higher overall yields than the synthesis described in the prior art. An improved multi-step synthesis of analogs. Numerous individual manufacturing steps and chemical intermediates differentiate this synthetic route from the routes described in the prior art. For example, this synthetic route relies on a novel configuration-specific synthesis of oxazoline intermediates and a unique stereoselective addition of formylamide to oxazoline.

The present invention is also directed to novel analogs of formulas VI and VII having unexpected biological activities. Lactacystin, crust-lactacystin β-lactone and their analogs have biological activity as inhibitors of the proteasome. Using them to treat conditions directly mediated by the function of the proteasome, such as muscle wasting, or indirectly mediated through proteins processed by the proteasome, such as the transcription factor NF-κB. Can be. The present invention relates to inhibiting proteasome function, or directly or indirectly mediated by proteasome function, by administering a compound of formula VI or VII having unexpectedly high activity in inhibiting proteasome. It is also directed to methods of treating certain conditions. In a preferred embodiment of the invention, a pharmaceutical composition comprising a compound of formula VI or VII is administered to treat ischemia or reperfusion injury. For example, in a preferred embodiment, the compounds can be used to treat, prevent or ameliorate post-stroke neuronal loss.

Synthetic Steps A first aspect of the present invention relates to a process for forming lactacystin having the formula VI below and its analogs and clast-lactacystin β-lactone having the formula VII below and its analogs:

Embedded image Wherein R ¹ is alkyl, alkenyl, alkynyl, cycloalkyl, aryl, alkaryl, or aralkyl (wherein any ring moiety of the aryl, aralkyl, or alkaryl may be optionally substituted R ² is alkyl, cycloalkyl, aryl, alkaryl, aralkyl, alkoxy, hydroxy, alkoxyalkyl, or amide, wherein any ring moiety of the aryl, aralkyl, or alkaryl is optionally substituted. R ⁷ is alkyl, aryl, aralkyl, or alkaryl (wherein any of the alkyl, aryl, aralkyl, or alkaryl may be optionally substituted)].

The process for forming these compounds is represented by formula V:

Embedded image Wherein R ¹ and R ² are the same as defined above for Formulas VI and VII. These steps include: (a) Formula I:

Embedded image Wherein R ¹ is the same as defined above, and R ³ is alkyl, alkenyl, alkynyl, cycloalkyl, aryl or alkaryl, wherein any of these is optionally substituted R ⁴ is aryl or heteroaryl (both of which may be optionally substituted)], and treating the substituted aryl or heteroaryl oxazoline with a strong base Or deprotonating the heteroaryl oxazoline to form an enolate; (b) transmetallating the enolate with a metal selected from the group consisting of titanium, aluminum, tin, zinc, magnesium and boron, followed by the formula XI
V:

Embedded image Wherein R ² is the same as defined above for formulas VI and VII; R ⁵ and R ⁶ are independently one of alkyl or alkaryl; or R ⁵ and R ⁶ is a 5- to 7-membered heterocyclic ring which, when taken together with the nitrogen atom to which they are attached, is optionally substituted and may optionally contain additional oxygen or nitrogen atoms. To form a compound of formula II:

Embedded image Wherein R ¹ to R ⁶ are as defined above; (c) catalytically hydrogenating the adduct of formula II to form an adduct of formula IV:

Embedded image Wherein R ¹ , R ² and R ³ are the same as defined above to form a γ-lactam represented by the formula: (d) saponifying the γ-lactam represented by the formula IV to obtain a compound represented by the formula V:

Embedded image [Wherein R ¹ and R ² are the same as defined above].

The carboxylic acid intermediate of formula V is cyclized by treatment with a cyclizing reagent to form a crust-lactacystin β-lactone of formula VII or an analog thereof, and optionally, This can be reacted with a thiol (R ⁷ SH), such as N-acetylcysteine, to form lactacystin having formula VI or an analog thereof.

Alternatively, the carboxylic acid intermediate of formula V can be directly coupled to a thiol (R ⁷ SH) such as N-acetylcysteine to form lactacystin having formula VI or an analog thereof. It can also be done.

A second aspect of the present invention provides a method comprising: (a) Formula VIII:

Embedded image Wherein, R ⁸ is isopropyl or benzyl] was deprotonated compound represented by, after which the resulting anion R ² CH ₂ C
Acylation with OCI provides a compound of formula IX:

Embedded image Wherein R ² and R ⁸ are the same as defined above; and (b) reacting the acyloxazolidinone of formula IX with benzyloxymethyl chloride in a stereoselective manner. And the formula X:

Embedded image Wherein R ² and R ⁸ are as defined above; (c) hydrolyzing the protected alcohol of formula X to form a formula XI:

Embedded image Wherein R ² is as defined above; (d) coupling the acid of formula XI with an amine R ⁵ R ⁶ NH ₂ to form a compound of formula XII :

Embedded image Wherein R ² , R ⁵ and R ⁶ are as defined above; (e) catalytic hydrogenation of an amide of formula XII to give a compound of formula XIII:

Embedded image Wherein R ² , R ⁵ and R ⁶ are as defined above; and (f) oxidizing the resulting alcohol of formula XIII to form a compound of formula XIV:

Embedded image Wherein R ² , R ⁵ and R ⁶ are the same as defined above, including the formation of an enantiomer-rich formyl amide of formula XIV.

In a third aspect of the present invention, there is provided (a) a compound of the formula XV:

Embedded image Asymmetric dihydroxylation of the alkene intermediate of formula XVIa:

Embedded image (B) reacting the optically active diol of formula XVIa with an orthoester derived from an aromatic carboxylic acid under an acid catalyst (Lewis or Bronsted acid) to form a mixed orthoester; Followed by the resulting mixed orthoester intermediate and a lower alkanoyl halide, hydrohalic acid
(HX, where X is a halogen), acid chlorides, and halogen-containing Lewis acids (eg, BBr ₃ , SnCl ₄ , Ti (OR) ₂ Cl ₂ , Ti (OR) ₃ Cl, and Me ₃ SiX (Where X is halogen or the like)) in the presence of a base with a reagent selected from the group consisting of:

Embedded image Wherein X is a halogen, preferably Cl, Br or I; (c) reacting the derivative of formula XVIIa with an alkali metal azide to form a compound of formula XVIIIa:

Embedded image (D) catalytic hydrogenation of the azide to form a compound of formula XIXa:

Embedded image (E) subjecting the compound of formula XIXa to ring closure conditions to form a substituted aryl or heteroaryl oxazoline of formula I with inversion of the configuration at the oxygen-substituted carbon. And formula Ia:

Embedded image Wherein R ¹ is alkyl, alkenyl, alkynyl, cycloalkyl, aryl, alkaryl, or aralkyl, wherein any ring portion of the aryl, aralkyl, or alkaryl is optionally substituted. R ³ is alkyl, cycloalkyl, aryl, or alkaryl (any of which may be optionally substituted); R ⁴ is aryl or heteroaryl (any of which is optionally substituted) Wherein, for each of formulas XV, XVIa, XVIIa, XVIIIa and XIXa, R ¹ , R ³ and R ⁴ are the same as defined above for formula I. -Forming an oxazoline comprising forming a trisubstituted cis-oxazoline compound of formula Ia. On.

Alternatively, a third aspect of the present invention provides a method comprising: (a) formula XV:

Embedded image Asymmetric dihydroxylation of the alkene intermediate of formula XVIb:

Embedded image (B) reacting the optically active diol of formula XVIb with an orthoester derived from an aromatic carboxylic acid under an acid catalyst (Lewis or Bronsted acid) to form a mixed orthoester And then the resulting orthoester intermediate and a lower alkanoyl halide, hydrohalic acid (HX, where X is a halogen), acid chloride, and a halogen-containing Lewis acid (eg, BBr ₃ , SnCl ₄ , Ti (O
R) ₂ Cl ₂ , Ti (OR) ₃ Cl, and a reagent selected from the group consisting of Me ₃ SiX (where X is a halogen, etc.) in the presence of a base to react with a reagent of formula XVI
Ib:

Embedded image Wherein X is a halogen, preferably Cl, Br or I; (c) reacting the derivative of formula XVIIb with an alkali metal azide to form a compound of formula XVIIIb:

Embedded image (D) catalytic hydrogenation of the azide to form a compound of formula XIXb:

Embedded image (E) subjecting the compound of Formula XIXb to ring closure conditions to form a substituted aryl- or heteroaryloxazoline of Formula Ib, wherein said ring closure reaction With the reversal of the configuration at the oxygen-substituted carbon
b to form a trans-oxazoline having the formula XV, XVI, X
A process for forming a trisubstituted trans-oxazoline compound of formula Ib, wherein for each of VII, XVIII and XIX, R ¹ , R ³ and R ⁴ are the same as defined above for formula I About.

For the above process, the following preferred values are applicable: R¹Is a preferred value of C_1-12Alkyl, especially C_1-8Alkyl, C_3-8Cycloalkyl, especially C_3-6Cycloalkyl, C_2-8Alkenyl, C_2-8Alkynyl, C_6-14 Aryl, especially C_6-10Aryl, C_6-10Ara (C_1-6) Lucil or C_1-6Al
Mosquito (C_6-10) Yl, wherein the aryl, aralkyl or alkaryl is
Any ring moiety may be optionally substituted). R¹Substituents which may be optionally present on the aryl ring of the moiety include one or more, preferably one or more,
Is two hydroxy, nitro, trifluoromethyl, halogen, C_1-6Alkyl, C_6-10Aryl, C_1-6Alkoxy, C_1-6Aminoalkyl, C_1-6Aminoalkoxy, amino, C_2-6Alkoxycarbonyl, carboxy, C_1-6Hydroxya
Luquil, C_2-6Hydroxyalkoxy, C_1-6Alkylsulfonyl, C_6-10Ally
Rusulfonyl, C_1-6Alkylsulfinyl, C_1-6Alkylsulfonamide, C _6-10 Arylsulfonamide, C_6-10Ara (C_1-6) Rukylsulfonamide, C_{1 -6} Alkyl, C_1-6Hydroxyalkyl, C_6-10Aryl, C_6-10Aryl (C_{1 -6} ) Alkyl, C_1-6Alkylcarbonyl, C_2-6Carboxyalkyl, cyano,
And trifluoromethoxy.

R¹Is more preferably a C such as ethyl, propyl or isopropyl_1-8 Alkyl; cycloalkyl such as cyclohexyl; or C such as phenyl _6-10 One of the aryls. Most preferably, it is isopropyl.

Preferred values for R ² are C _1-8 alkyl, C _3-8 cycloalkyl, especially C _3-6 cycloalkyl, C _1-8 alkoxy, C _2-8 alkenyl, C _2-8 alkynyl, C _2-8 alkynyl, _6-14 aryl, especially C _6-10 aryl, C _6-10 ar (C _1-6 ) alkyl, or C _1-6 alka (
C _6-10 ) yl, wherein the ring moiety of any of the aryl, aralkyl, or alkaryl may be optionally substituted with any of the substituents defined above for R ^1. .

[0030] R ² is more preferably methyl, ethyl, or a C _{1 -4} alkyl, such as propyl or butyl; or a such as methoxy or ethoxy C _1-4 alkoxy. Most preferred are methyl, ethyl, propyl and butyl.

For R ³ , various ester functions can be used at this moiety. Preferred values are C _1-8 alkyl, C _3-8 cycloalkyl, especially C _4-7 cycloalkyl, C _2-8 alkenyl, C _2-8 alkynyl, C _6-14 aryl, especially C _6-10 Aryl,
C _6-10 aralkyl (C _1-6 ) alkyl, or C _1-6 alk (C _6-10 ) aryl, wherein any of these may be optionally substituted. Substituents which may exist optionally on R ^3, 1 or greater than that of said definitions of substituents for R ^1, preferably included one or Two.

[0032] R ³ is preferably C _1-4 alkyl, C _6-10 aryl or C _6-10 Ala, (C _{1- 6)} alkyl. Most preferred are methyl, ethyl, tert-butyl and benzyl.

R ⁴ is preferably C _6-10 aryl, preferably phenyl, or thienyl, benzo [b] thienyl, furyl, pyranyl, isobenzofuranyl, benzoxazolyl, 2H-pyrrolyl, pyrrolyl, imidazolyl, Pyrazolyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, indolizinyl, isoindolyl, 3H
-A heteroaryl group selected from the group consisting of indolyl, indolyl, indazolyl, purinyl, 4H-quinolizinyl, isoquinolyl, quinolyl or triazolyl. The phenyl or heteroaryl group may be optionally substituted by one or two of the substituents defined above for R ¹ . Most preferred are phenyl and phenyl substituted with halogen, C _1-6 alkyl, C _1-6 alkoxy, carboxy, amino, C _1-6 alkylamino and / or di (C _1-6 ) alkylamino. .

R ⁵ and R ⁶ are independently one of alkyl, aralkyl or alkaryl; alternatively, R ⁵ and R ⁶ , when taken together with the nitrogen atom to which they are attached, Forming a 5- to 7-membered heterocyclic ring which is optionally substituted and may optionally contain additional oxygen or nitrogen atoms. Optional substituents are those listed above for R ¹ .

R^FiveAnd R⁶Is preferably C_1-6Alkyl, C_6-10Ara (C_1-6) Lucil
Or C_1-6Arca (C_6-10) Is, or together with, the nitrogen atom to which they are attached, optionally substituted and optionally further substituted
Forming a 5- to 7-membered heterocyclic ring which may contain an oxygen atom or a nitrogen atom
You. NR^FiveR⁶The most preferred values for are dimethylamino, diethylamino,
Pyrrolidino, piperidino, morpholino, oxazolidinone, and halogen, C _1-6 Alkyl, C_6-10Ara (C_1-6) Luquil, C_1-6Oxazolidinones substituted with alkoxy, carboxy, and / or amino.

R ⁷ is preferably C _1-8 alkyl, C _3-8 cycloalkyl, C _6-10 aryl, C _6-10 ar (C _1-6 ) alkyl, or C _1-6 alk (C _6-10 ) Ryl (any of these may be optionally substituted). Substituents which may be optionally present on the Izu Re or both rings or chain portion of R ^7, 1 or greater than that of the substituents described above for R ^1, preferably 1 or 2 substituents Groups are included. Preferably, R ⁷ is that it together with the sulfur atom to which they are attached cysteine or N- acetylcysteine, a cysteine derivatives such as such as glutathione.

Reaction Scheme 1 shows lactacystin and crust-substitution from substituted oxazoline starting materials.
1 is a general reaction scheme for forming a lactacystin β-lactone analog.

[0038]

Embedded image

The starting oxazoline I, which can be in either the cis (Ia) or trans (Ib) configuration, is deprotonated with a strong base to form an enolate. An example of a suitable base for use in this reaction is lithium diisopropylamide (LDA)
, Lithium tetramethylpiperidide (LiTMP), lithium, sodium or potassium hexamethyldisilazide (LiHMDS, NaHMDS, KH
A constrained amide base such as MDS); and an organic base containing a constrained alkyl lithium reagent such as sec-butyllithium and tert-butyllithium. The reaction is preferably performed at low temperature in an ethereal solvent such as diethyl ether, tetrahydrofuran (THF) or dimethoxyethane (DME). The reaction temperature is preferably about -100C to about -30C, more preferably -85C to -5C.
0 ° C, most preferably in the range of -85 ° C to -75 ° C. This reaction temperature is important in determining the stereochemical consequences of the subsequent addition to the aldehyde, but at lower temperatures better selectivity is obtained.

[0040] Following the deprotonation step, the enolate is transmetallated with a metal selected from the group consisting of titanium, aluminum, tin, zinc, magnesium and boron. This is step per preferred reagent, titanium or aluminum Lewis acid, for example, Me ₂ AlCl, or (i-PrO) ₃ TiCl or mixtures of the two. Preferably, 1 to 3 molar equivalents of Lewis acid are used, more preferably 2 to 3 equivalents, most preferably about 2.2 to 2.3 equivalents. Subsequent treatment of the enolate with formylamide (XIV) gives the adduct II. Excess aldehyde was washed off with sodium bisulfite solution and the crude material was carried on to the next step without further purification. By using 2.2-2.3 equivalents of Me ₂ AlCl, the (6S) -product (
Lactacystin numbering), while the use of one equivalent of Me ₂ AlCl results in the selective formation of (6R) -product in a ratio of about 5: 1.

The catalytic hydrogenation hydrolysis of the adduct II gives, as a mixture of (6S)-and (6R) -epimers, sometimes aminodiols III:

Embedded image To obtain the desired γ-lactam (IV).

[0042] Catalysts useful for this reaction include palladium black, palladium on activated carbon, palladium hydroxide on carbon, and the like. Organic solvents suitable for use in this reaction include lower alkanols such as methanol, ethanol or isopropanol, lower alkanoates such as ethyl acetate, lower alkanoic acids such as acetic acid, or mixtures thereof. The reaction is carried out under a hydrogen atmosphere at about 15 to about 100 psi. ,
More preferably, from about 30 to about 50 psi. It is performed at a pressure in the range. Alternatively, the transfer hydrogenation method (R.A.W. Johnstone et al., Chem. Rev.
85: 129 (1985)), wherein adduct II is treated with a catalyst and a hydrogen donor at atmospheric pressure.

Upon heating of the crude product mixture, the aminodiol III is converted to γ-lactam IV, which can be isolated from II in almost 60-75% overall yield. The heating step is conveniently carried out by first filtering off the catalyst used in the hydrogenation step and then heating and refluxing the filtrate. Aminodiol I in the crude product mixture
If no II is present, the heating step is omitted. The cyclization following saponification of the ester gives β-lactone VII in a yield of 40-90%, generally more than 60%. Cyclization includes arylsulfonyl chloride, benzotriazol-1-yloxytris (dimethylamino) phosphonium hexafluorophosphate (BOP reagent), O- (1H-benzotriazol-1-yl) -N,
N, N ', N'-tetramethyluronium tetrafluoroborate (TBTU)
, Alkyl chloroformates, aryl chloroformates, or alkenyl chloroformates, and the like. Isopropenyl chloroformate is the preferred reagent for this step. Because
Because all by-products are volatile, this product does not require chromatographic purification.

Crust-lactacystin β-lactone was prepared using a reported procedure (Core
y et al., Tetrahedron Lett. 34: 6977 (1993))
Can be converted to lactacystin by treating β-lactone with N-acetylcysteine according to The reaction proceeds similarly when β-lactone VII is reacted with another thiol. Alternatively, the lactacystin analog can be prepared by coupling the carboxylic acid intermediate V with a thiol to form the corresponding thiol ester VI
It is also prepared by forming Thus, the methods of the present invention are useful for the synthesis of both lactacystin and crust-lactacystin β-lactone, and analogs thereof.

The enantiomer-rich formylamide XIV used in the aldol reaction is new. They can be prepared according to a typical reaction sequence such as that shown in Reaction Scheme 2. For the purposes of the present invention, the term "enantiomer-rich" means that one enantiomer is present in excess of another;
That is, one enantiomer is present in greater than 50% of the mixture. The term "configuration-selective" is intended to mean that one enantiomer or diastereomer is formed in excess of another enantiomer or other diastereomer (s) by a synthesis or reaction step. Used for

[0046]

Embedded image

(S)-(−)-4-benzyl-2-oxazolidinone (VIIIa) or
(S)-(-)-4-isopropyl-2-oxazolidinone (VIIIb) (formula
Medium, R⁸Acylation of the anion of benzyl or isopropyl) affords the acyloxazolidinone IX in greater than 80% yield. Continuing configuration
Selective benzyloxymethylation (Evans et al., J. Am. Chem. S.
oc. 112: 8215 (1990)) to provide protected 80% yields.
Coal X is obtained, except that benzyl chloromethyl ether is freshly prepared
(Organic Synthesis 52:16 (1 by Connor et al.)
974)). Peroxide-mediated hydrolysis affords acid XI, which is reacted with amine
Coupling affords amide XII in an overall yield generally greater than 50%.
You. For the oxidation of the resulting alcohol (XIII) following the hydrogenolysis of the benzyl group
Thus, formylamide XIV is obtained in a yield of 80 to 85%. Pearlman catalyst
(Pd (OH)_Two) Is preferably used in the hydrocracking step. The final oxidation step is described in Dess and Martin, J. et al. Org. Chem. 48: 4156 (1983
), Or 2,2,6,6-tetramethyl-
Buffer in the presence of piperidinyloxy (TEMPO) free radical and bromide ion
Works best with hydrochloride hydrochloride (J.
Org. Chem. 50: 4888 (1985); Org. Synth. Col
l. 8: 367 (1993)). Tetrapropyl-ammonium perruthenate
Other mild oxidants such as (TPAP) can also be used. Formylamide
XIV reduces the aldehyde with sodium borohydride and gives the alcohol
With R-(+)-α-methoxy-α- (trifluoromethyl) phenylacetyl chloride
Conversion to the corresponding Mosher ester using
Can be shown to be enantiomerically pure (Dale et al.
J. Org. Chem. 34: 2543 (1969)). At 300 MHz ¹ H-NMR analysis reveals a single diastereomer. The aldehyde prepared according to Reaction Scheme 2 is sterically stable and can be stored at 0 ° C. for one week.
Even afterwards it shows no signs of enantiomeric degradation. The aldehyde is
It is sterically stable under the conditions of the Dole reaction, and the adduct II has C (7)
Substituent R^TwoFormed without epimerization of

The synthetic method will work with any substituent of R ¹ that is strong base and stable to hydrogenation. Isopropyl is a preferred substituent for good proteasome inhibitory activity of the final product.

[0049]

Embedded image

The present invention also relates to a new route for forming oxazoline starting material I.
The overall synthesis involves five steps (Scheme 3), and the cis-substituted oxazoline Ia
Which is then used in the method described above. The first step shown in Reaction Scheme 3 is
Sharpless asymmetric dihydroxylation of alkene XV (J. Org. Chem. 57: 2768 (1992) by Sharless et al .; C. by Kolb et al.).
hem. Rev .. 94: 2483 (1994); Shao and Goodman.
J. Org. Chem. 61: 2582 (1996)). If not commercially available, alkene XV is prepared by Wittig condensation between aldehyde and carbethoxymethylene triphenylphosphorane (Tetrahedron 50: 9181 by Hale et al. (1994)). Other olefination techniques are known in the art. The dihydroxylation reaction is preferably
AD-mix-β (Aldrich Chemi) in the presence of methanesulfonamide
cal Co. Using Sharpless's face-selection rule (S
As expected by harless face-selection rule), the diol XVIa is obtained in a configurationally selective manner. On a large scale, the dihydroxylation reaction is preferably performed using N-methylmorpholine-N-oxide (NMO) as reoxidant instead of K ₃ Fe (CN) ₆ present in AD-mix-β. . Although proceeding with somewhat lower enantiomeric selectivity, this approach allows for a more concentrated reaction mixture and greatly simplifies work-up. The enantiomeric purity of the product can be increased by recrystallization.

In the next step, the diol XVIa is catalyzed under a Lewis or Bronsted acid catalyst.
Is treated with an orthoester to give a mixed orthoester which is converted in situ to the haloester XVIIa by treatment with an acyl halide (Tetrahedron Lett. 37: 4525 (Haddadd et al.)
1996)). Acyl halides, particularly acetyl halides, are preferred for this reaction, but other acid halides such as HCl, HBr, HI, Me ₃ SiCl, Me ₃ SiI, Me ₃ SiBr, etc. can also be used. BBr ₃ , SnCl ₄ ,
_{Ti (OR) 2 Cl 2,} Ti (OR) 3 using a halogen-containing Lewis acids represented by the formula ML _n X, such as such as Cl can be. In the above formula, M is B, Ti, Sn, Al
L is any ligand suitable for the metal, preferably an alkoxide or halogen group;
n is an integer that results in a stable complex; X is halogen. Preferably, acetyl bromide is used to produce the haloester XVIIa. Preferably, the orthoester used in this reaction is derived from an aromatic or heteroaromatic carboxylic acid. More preferably, the orthoester is derived from a benzoic acid, for example, trimethylorthobenzoic acid. It is preferred to use a boron trifluoride ether compound such as a Lewis acid catalyst in forming the mixed orthoester, but other acids such as HBr, SnCl ₄ , TiCl ₄ , BBr ₃ can be used.

After work-up, the crude halide XVIIa is treated with an alkali metal azide in a polar aprotic solvent such as dimethylsulfoxide (DMSO) or N, N-dimethylformamide (DMF) to give the azide. XVII
Is converted to Ia. Catalytic hydrogenation of azide XVIIIa over a palladium catalyst in ethyl acetate was accomplished by simultaneous transfer of aroyl groups (Wang et al., J. Org. Chem.
59: 5014 (1994)) to give the hydroxyamide XIXa.

Treatment of XIXa with thionyl chloride in methylene chloride affects ring closure with inversion of configuration at the hydroxyl-substituted carbon atom to produce the cis-substituted oxazoline starting material Ia. Other reagents suitable for use in this reaction include sulfuryl chloride, phosphorus trichloride, phosphorus oxychloride, and (methoxycarbonylsulfamoyl) -triethylammonium hydroxide, inner salt (Burgess reagent). Treatment of XIXa under Mitsunobu conditions (Synthesis by Mitsunobu: 1 (1981)) will also affect ring closure. The oxazoline ring oxygen atom is intended to be the C (9) -hydroxyl group in the final products VI and VII. Under equilibrium conditions (sodium methoxide, methanol), the cis-oxazoline (Ia) is converted to trans-oxazoline (Ib) by inversion of the configuration of the ester substituent, while the configuration of the R ¹ substituent remains fixed. You. The cis- and trans-oxazolines can both be used in the manner shown in Scheme 1 with equivalent results.

In an alternative route for forming oxazoline starting material 1, p-toluenesulfonic acid (p-TsOH) is used to effect ring closure (Scheme 4). In this case, ring closure proceeds while maintaining the configuration at the hydroxyl-substituted carbon atom to give trans-oxazoline (1b). C of final product (
To obtain the appropriate stereochemistry in 9), the chiral ligand used in the dihydroxylation reaction must be chosen to provide the opposite face selectivity to that shown in Scheme 3. For example, AD-mix-α is used instead of AD-mix-β. All other steps in the sequence proceed similar to those described for the synthesis of cis-oxazoline Ia.

[0055]

Embedded image

Compounds Many of the compounds described above are novel compounds; new compounds are also claimed. The fourth, fifth and sixth aspects of the present invention relate to lactacystin analogues, which can be produced by the synthetic routes described herein; pharmaceutical compositions containing such compounds; and A method of treating a subject having a condition mediated by a protein processed by the proteasome by administering to the subject an effective amount of the disclosed pharmaceutical composition. These methods include Alzheimer's disease, cachexia, cancer, inflammation (eg, allergy, inflammatory response associated with bone marrow or solid tissue transplantation, or, but not limited to, arthritis, multiple sclerosis, inflammatory Disease states including intestinal diseases and parasitic diseases such as malaria);
Includes the treatment of psoriasis, restenosis, seizures, and myocardial infarction.

The compounds of formulas VI and VII disclosed herein are highly selective for the proteasome and trypsin, α-chymotrypsin, calpain I
Does not inhibit other proteases such as, calpain II, papain and cathepsin B.

[0058] Fentany et al. (WO 96/32), which is expressly incorporated herein by reference.
As disclosed by US Pat. No. 105), lactacystin, crust-lactacystin β-lactone and their analogs have biological activity as inhibitors of the proteasome. They can be used to treat conditions that are directly mediated by the function of the proteasome, such as muscle wasting, or indirectly through proteins that are processed by the proteasome, such as the transcription factor NF-κB. it can. The compounds prepared by the methods of the present invention can also be used to determine whether a cellular, developmental or physiological process or result is regulated by proteasome proteolytic activity.

These compounds with unexpected proteasome function-inhibitory activity are represented by formulas VI and VII:

Embedded image Wherein R ¹ is C _1-12 alkyl, C _3-8 cycloalkyl, C _2-8 alkenyl, C _2-8 alkynyl, C _6-14 aryl, C _6-10 ar (C _1-6 ) alkyl , or a C _1-6 alk (C _{6- 10)} Lil; R ² is a C _2-6 alkyl; R ⁷ is C _1-8 alkyl, C _3-8 cycloalkyl, C _6-10 aryl , C _6-10 aralkyl (C _1-6 ) alkyl, or C _1-6 alk (C _6-10 ) aryl, wherein any of these may be optionally substituted. Or a salt thereof. Substituents which may optionally be present on either or both the ring or chain portion of R ⁷ include one or more, preferably one or two, of the substituents defined above for R ^1. It is.

Preferred compounds are those in which R ¹ is C _1-4 alkyl, more preferably isopropyl. R ² is preferably ethyl, n- propyl, n- butyl or isobutylene chill. Preferably, R ⁷ together with the sulfur atom to which it is attached, becomes cysteine or N- acetylcysteine, cysteine derivatives such as such as glutathione.

A seventh aspect of the present invention provides a compound of formula XIV:

Embedded image Wherein R ² is C _1-8 alkyl, C _3-8 cycloalkyl, C _2-8 alkenyl, C _2-8 alkynyl, C _6-14 aryl, C _6-10 ara (C _1-6 ) alkyl or C _1-6 alk (C _{6- 10)} a drill,; R ⁵ and R ⁶ are independently, C _1-6 alkyl, C _6-10 Ara (C _1-6) alkyl or, C _1-6 alk (C _6-10 ) lyl, or together with the nitrogen atom to which they are attached, may be optionally substituted and optionally contain further oxygen or nitrogen atoms. Forms an optionally 5- to 7-membered heterocycle
] Or a salt thereof. Preferred compounds are those where R ² is C _2-6 alkyl.

An eighth aspect of the present invention provides a compound of formula II and III:

Embedded image Wherein R ¹ is alkyl, alkenyl, alkynyl, cycloalkyl, aryl,
R ² is alkyl, cycloalkyl, aryl, alkaryl, aralkyl, alkoxy, hydroxy, , alkoxyalkyl or an amide (where the aryl, any ring moiety of the aralkyl or alkaryl may also be optionally substituted),; R ³ is alkyl, alkenyl, alkynyl, cycloalkyl, aryl, or alkaryl (here, may be optionally substituted none of them); R ⁴ is a heteroaryl optionally substituted by aryl or optionally substituted optionally; R ⁵ and R ^6, independently, alkyl or alkali Which is one one of; or, R ⁵ and R ^6, if they are taken together with the nitrogen atom to which they are attached
To form a 5- to 7-membered heterocyclic ring which may be optionally substituted and optionally further contain an oxygen or nitrogen atom, or a salt thereof. . The most preferred values for NR ⁵ R ⁶ is dimethylamino, diethylamino, pyrrolidino, piperidino, morpholino, oxazolidinones, and halogen, C _1-6 alkyl, C _6-10 Ara (C _{1 -6)} alkyl, C _1-6 alkoxy Oxazolidinone substituted with carboxy and / or amino], or a salt thereof.

Preferred compounds of formulas II and III are:¹But C_1-12Alkyl, C_3-8Cycloalkyl, C_2-8Alkenyl, C_{Two -8} Alkynyl, C_6-14Aryl, C_6-10Ara (C_1-6) Lucil or C_1-6Al
Mosquito (C_6-10) Yl, wherein the aryl, aralkyl or alkaryl is
Any ring moiety may be optionally substituted); R^TwoBut C_1-8Alkyl, C_3-8Cycloalkyl, C_2-8Alkenyl, C_2-8Alkynyl, C_6-14Aryl, C_6-10Ara (C_1-6) Lucil or C_1-6Arca (C _6-10 ) Is ril (wherein any of the aryl, aralkyl or alkaryl)
May also be optionally substituted); R^ThreeBut C_1-8Alkyl, C_3-8Cycloalkyl, C_2-8Alkenyl, C_2-8Alkynyl, C_6-14Aryl, C_6-10Ara (C_1-6) Lucil or C_1-6Arca (C _6-10 ) Yl, wherein any of these may be optionally substituted
); R^FourIs optionally substituted C_6-10Aryl, or thienyl, benzo [β] thienyl, furyl, pyranyl, isobenzofuranyl, benzoxazo
Ryl, 2H-pyrrolyl, pyrrolyl, imidazolyl, pyrazolyl, pyridyl, pyra
Dinyl, pyrimidinyl, pyridazinyl, indolizinyl, isoindolyl, 3H
-Indolyl, indolyl, indazolyl, purinyl, 4H-quinolizinyl, i
Optionally selected from the group consisting of soquinolyl, quinolyl and triazolyl
An optionally substituted heteroaryl group; R^FiveAnd R⁶But independently C_1-6Alkyl, C_6-10Ara (C_1-6) Lucil
Or C_1-6Arca (C_6-10) Is lyl or, together with the nitrogen atom to which they are attached, optionally substituted and optionally
Forming a 5- to 7-membered heterocyclic ring which may contain an element atom or a nitrogen atom
Is the thing. NR^FiveR⁶The most preferred values for are dimethylamino, diethyl
Amino, pyrrolidino, piperidino, morpholino, oxazolidinone, and ha
Rogen, C_1-6Alkyl, C_6-10Ara (C_1-6) Luquil, C_1-6Oxazolidinones substituted with alkoxy, carboxy and / or amino.

A ninth aspect of the invention is a compound of formula XVIIa, XVIIb, XVIIIa, XVII
Ib, XIXa or XIXb:

Embedded image Wherein R ¹ is alkyl, alkenyl, alkynyl, cycloalkyl, aryl, alkaryl, or aralkyl, wherein any ring portion of the aryl, aralkyl, or alkaryl is optionally substituted. R ³ is alkyl, alkenyl, alkynyl, cycloalkyl, aryl, or alkaryl (wherein any of these may be optionally substituted); R ⁴ is optionally substituted Aryl or an optionally substituted heteroaryl], or a salt thereof.

Preferred compounds of formula XVII, XVIII or XIX are those wherein R¹But C_1-12Alkyl, C_3-8Cycloalkyl, C_2-8Alkenyl, C_{Two -8} Alkynyl C_6-14Aryl, C_6-10Ara (C_1-6) Lucil or C_1-6Arca
(C_6-10) Yl, wherein said aryl, aralkyl or alkaryl is
The offset ring moiety may also be optionally substituted);^ThreeBut C_1-8Alkyl, C_3-8Cycloalkyl, C_2-8Alkenyl, C_2-8Alkynyl, C_6-14Aryl, C_6-10Ara (C_1-6) Lucil or C_1-6Arca (C _6-10 ) Yl, wherein any of these may be optionally substituted
I); R^FourIs optionally substituted C_6-10Aryl or thienyl, benzo [b] thienyl, furyl, pyranyl, isobenzofuranyl, benzoxazo
Ryl, 2H-pyrrolyl, pyrrolyl, imidazolyl, pyrazolyl, pyridyl, pyra
Dinyl, pyrimidinyl, pyridazinyl, indolizinyl, isoindolyl, 3H
-Indolyl, indolyl, indazolyl, purinyl, 4H-quinolizinyl, i
Optionally selected from the group consisting of soquinolyl, quinolyl and triazolyl
A heteroaryl group which may be substituted.

Definitions As used herein, the term “alkyl” includes methyl, ethyl, propyl, isopropyl, butyl, t-butyl, isobutyl, pentyl, hexyl, isohexyl, 1-ethylpropyl, heptyl, 4,4 Includes both straight-chain and branched radicals of up to 12, preferably 1-8 carbons, such as dimethylpentyl, octyl, 2,2,4-trimethylpentyl, nonyl, decyl, undecyl and dodecyl .

The term “substituted alkyl” as used herein includes one, two or three halo,
Hydroxy, nitro, trifluoromethyl, halogen, C _1-6 alkyl, C _6-10 aryl, C _1-6 alkoxy, C _1-6 aminoalkyl, C _1-6 aminoalkoxy, amino, C _2-6 alkoxycarbonyl , Carboxy, C _1-6 hydroxyalkyl,
C _2-6 hydroxyalkoxy, C _1-6 alkylsulfonyl, C _6-10 arylsulfonyl, C _1-6 alkylsulfinyl, C _1-6 alkylsulfonamide, C _6-10 arylsulfonamide, C _6-10 ara ( C _1-6 ) alkylsulfonamide, C _1-6 alkyl, C _1-6 hydroxyalkyl, C _6-10 aryl, C _6-10 aryl (C _1-6 ) alkyl, C _1-6 alkylcarbonyl, C _{2 -6} Carboxyalkyl, cyano and trifluoromethoxy and / or alkyl groups as defined above with a carboxy substituent are included.

The term “cycloalkyl” as used herein includes saturated cyclic hydrocarbons containing 3 to 12, preferably 3 to 8, carbons, including any group Cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclooctyl, cyclodecyl and cyclododecyl, optionally substituted by substituents such as halogen, C _1-6 alkyl, C _1-6 alkoxy and / or hydroxy groups. included.

The term “heteroaryl” as used herein refers to 5 to 14 ring atoms, preferably 5, 6, 9 or 10 ring atoms; 6, 10 or 14π electron; and a group having a carbon atom and one, two or three oxygen, nitrogen or sulfur heteroatoms (examples of a heteroaryl group are: thienyl,
Benzo [b] thienyl, naphtho [2,3-b] thienyl, thianthrenyl, furyl, pyranyl, isobenzofuranyl, benzoxazolyl, clomethinyl, xanthenyl, phenoxathiinyl, 2H-pyrrolyl, pyrrolyl, imidazolyl, pyrazolyl , Pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, indolizinyl,
Isoindolyl, 3H-indolyl, indolyl, indazolyl, purinyl, 4
H-quinolizinyl, isoquinolyl, quinolyl, phthalazinyl, naphthyridinyl,
Tetrazolyl, quinazolinyl, cinnolinyl, pteridinyl, 4αH-carbazolyl, carbazolyl, β-carbolinyl, phenanthridinyl, acridinyl,
(Perimidinyl, phenanthrolinyl, phenazinyl, isothiazolyl, phenothiazinyl, isoxazolyl, furazanyl, and phenoxazinyl groups)
.

The term “aryl” as used herein, by itself or as part of another group, refers to 6 to 12 carbons in the ring moiety, such as phenyl, naphthyl or tetrahydronaphthyl, preferably 6 Refers to a monocyclic or bicyclic aromatic group containing -10 carbons in the ring portion.

As used herein, the term “aralkyl” or “arylalkyl”, by itself or as part of another group, refers to a compound having an aryl substituent, such as benzyl, phenylethyl or 2-naphthylmethyl. Refers to a C _1-6 alkyl group as discussed above.

The term “alkaryl” or “alkylaryl” as used herein, by itself or as part of another group, refers to a C _1-6 alkyl substituent such as toluyl, ethylphenyl, or methylnaphthyl. Refers to an aryl group as discussed above having

The term “optionally substituted” when used in reference to an aryl, aralkyl, alkaryl or 5-, 6-, 9- or 10-membered heteroaryl group means that the ring portion of the group is C _1-6 alkyl, C _3-8 cycloalkyl, C _1-6 alkyl (C _3-8 ) cycloalkyl, C _2-8 alkenyl, C _2-8 alkynyl, cyano, amino, C _1-6 alkyl Amino, di (C _1-6 ) alkylamino, benzylamino, dibenzylamino, nitro, carboxy, carbo (C _1-6 ) alkoxy, trifluoromethyl, halogen, C _1-6 alkoxy, C _6-10 aryl, C _6-10 aryl (C _{1 -6)} alkyl, C _6-10 aryl (C _1-6) alkoxy, hydroxy, C _1-6 alkylthio, C _1-6 alkylsulfinyl, C _1-6 alkylsulfonyl, C _{6- 10} arylthio, C _6-10 A Rusurufiniru, C _6-10 arylsulfonyl, C _6-10 aryl, C _1-6 alkyl (C _6-10) aryl, and halo (C _6-10) 1 or 2 substituents independently selected from aryl Means that they may be optionally substituted by groups.

The term “alkoxy” refers to an alkyl group as described above bonded to oxygen. The terms "halogen" or "halo" as used herein, by themselves or as part of another group, refer to chlorin, bromine, fluorin or iodine. The term "amide" as used herein refers to formylamino, alkylcarbonylamino or arylcarbonylamino.

Uses Pharmacological data for the crust-lactacystin β-lactone analog prepared by the method of the present invention are listed in Table 1. These compounds are all irreversible inactivators of the 20S proteasome, which acylate the N-terminal threonine residue of the X / MB1 subunit. The value K _obs / [I] is a measure of the rate of enzyme inactivation. Some compounds show an improved activity, ie, a faster rate of inactivation when compared to crust-lactacystin β-lactone itself (2). The most potent compounds in this enzyme assay are
This is a 7-methoxy derivative 3f. However, 3f is less potent than 2 when assayed in cell culture.

The lactone ring is subject to nucleophilic attack by water as well as threonine residues of the proteasome X / MB1 subunit. Hydrolysis results in the formation of hydroxy acid V, which is not active as an inhibitor of the proteasome. The relative potency in cell culture is a complex of many factors, including enzyme potency, cell permeability, and rate of hydrolysis. Although more potent for the enzyme than 2, 3f hydrolyzes more rapidly in cell culture, resulting in much weaker activity. In contrast, analogs 3a-3d surprisingly show improved potency in enzyme assays as well as in cell culture.

Using the disclosed compounds, conditions mediated directly by the proteolytic function of the proteasome, such as muscle wasting, or indirectly mediated by proteins processed by the proteasome, such as NF-κB Treat the symptoms. The proteasome is involved in the rapid clearance and post-translational processing of proteins involved in cell regulation (eg, cell cycle, gene transcription, and metabolic pathways), intercellular communication and immune responses (eg, antigen presentation). Specific examples include β-amyloid proteins and regulatory proteins such as cyclins and the transcription factor NF-κB. As used herein, treatment includes reversing, reducing, or preventing symptomatic symptoms, clinical signs, and existing pathologies, in a manner ameliorating or stabilizing a subject's symptoms. It is.

Another embodiment of the present invention relates to cachexia and muscle wasting disease. The proteasome degrades many proteins in mature reticulocytes and growing fibroblasts. In cells taken from insulin or serum, the rate of protein hydrolysis is almost doubled.

Inhibition of the proteasome reduces protein hydrolysis, thereby reducing both muscle protein loss and nitrogen load to the kidney or liver.
Proteasome inhibitors are useful for treating conditions such as cancer, chronic infections, fever, muscle wasting (atrophy) and denervation, nerve damage, fasting, renal failure associated with acidosis, and liver failure . See, for example, US Patent No. 5,340,736 to Goldberg (1994).

Thus, embodiments of the present invention include methods of reducing the rate of muscle protein degradation in cells and methods of reducing the rate of intracellular protein degradation. Each of these methods includes contacting a cell (in vivo or in vitro, eg, a muscle of a subject) with an effective amount of a compound of the formula disclosed herein (eg, a pharmaceutical composition). Is included.

Proteasome inhibitors block the processing of ubiquitinated NF-κB in vitro and in vivo. Proteasome inhibitors also inhibit IκB-α degradation and NF-κB activation (Palombell
EMBO J. a .; and Traenckner et al. 13: 5433-5
441 (1994)). One embodiment of the present invention is a method of inhibiting IκB-α degradation, comprising contacting a cell with a compound of the formula described herein. Further embodiments include contacting a cell, muscle, organ, or subject with a compound of the formulas described herein to determine the cellular content of NF-κB in the cell, muscle, organ, or subject. It is a method of lowering. In certain embodiments, administration of a compound of the formula disclosed herein results in allergy, an inflammatory response associated with bone marrow or solid organ transplantation, or, but not limited to, arthritis, Methods for treating disease states, including inflammatory bowel disease, asthma, and multiple sclerosis. A preferred embodiment of the present invention is directed to treating asthma by administering a compound of formula VI or formula VII, most preferably of formula 3b.

The proteasome inhibitor was obtained from Bra, filed February 17, 1998.
US Patent Application No. (ProScript Doc
No. 08 / 988,339 filed on Dec. 3, 1997 and U.S. Patent Application No. 08 / 801,936 filed on Feb. 15, 1998. It is also useful for the treatment of ischemia or reperfusion injury, particularly for preventing or reducing the size of infarcts following vaso-occlusion, such as occurs during a stroke or heart attack, as described in is there. Proteasome inhibitors also block the proteasome-dependent metamorphosis of strict animal parasites (Gonzalez et al., J. Exp. Med. 184: 1909 (
1996)). Accordingly, a further embodiment of the present invention includes a method of treating infarction or a protozoan parasitic disease, comprising administering a compound of the formulas disclosed herein. In a preferred embodiment of the invention, formula VI or formula VII
Is administered to prevent or reduce infarction following vasculature obstruction. The compound can be administered from about 0 to about 10 hours after the onset of an attack to treat or reduce nerve loss following an ischemic event. Compound 3b is the most preferred compound in this aspect of the invention.

[0083] Proteasome inhibitors also block the degradation of cell cycle regulatory proteins, such as cyclins and cyclin-dependent kinase inhibitors, and tumor suppressor proteins, such as p53. Thus, another embodiment of the present invention is:
Includes methods for using the compounds of the formulas described herein to block the cell cycle and treat cell proliferative diseases such as cancer, psoriasis, and restenosis.

The term “inhibitor” is meant to describe a compound that blocks or reduces the activity of an enzyme (eg, the proteasome, or the X / MB1 subunit of the 20S proteasome). Inhibitors are competitive, uncompetitive (uncomp
active) or non-competitive inhibition. Inhibitors can bind reversibly or irreversibly, and thus the term also includes compounds that are suicide substrates of the enzyme. The inhibitor may modify one or more sites above or near the active site of the enzyme, or in other cases it may cause a conformational change on the enzyme.

The amount and mode of administration of the protease inhibitors and compositions of the present invention
It can be easily determined by those skilled in the art. In general, the dosage of the composition of the invention will depend on the type of composition used; age; health; the medical condition to be treated; Nature; degree of tissue damage;
Gender; duration of symptoms; and response indicators, if any, will vary depending on other variables to be adjusted by the individual physician. The desired dose can be administered in one or more applications to achieve the desired result. Pharmaceutical compositions containing the proteasome inhibitors of the present invention can be provided in unit dosage form.

Compositions within the scope of the present invention include all compositions that contain a compound of the present invention in an amount effective to achieve its intended purpose. While individual needs vary, determination of optimal ranges of effective amounts of each component is within the skill of the art. Typically,
The compound may be administered to a mammal, e.g., a human, to treat a proteasome-mediated condition, such as seizures or asthma, in a daily dose of 0.0025-50 mg / kg of the mammal's body weight, or a significant amount of its pharmaceutically acceptable. Can be administered. For intramuscular injection, the dose is generally about 1/2 of the oral dose.

In a method of preventing or reducing the size of an infarction, the compound is administered by intravenous injection at a dose of about 0.01 to about 10 mg / kg, preferably about 0.025 to about 1 mg / kg. Can be administered.

A unit oral dose may contain about 0.01 to about 50 mg, preferably about 0.1 to about 10 mg.
Can be included. The unit dose can be administered as one or more tablets each containing from about 0.1 to about 10, conveniently about 0.25 to 50 mg of the compound or solvate thereof, one or more times per day. can do. For use in treating stroke, it is preferable to administer a single dose from 0 to about 10 hours after the event, preferably from 0 to about 6 hours after the event.

The following examples are illustrative, but not limiting, of the methods and compositions of the present invention. Other suitable modifications and applications of the various conditions and indications that are routinely cited and obvious to those skilled in the art are within the spirit and scope of the invention.

The preparation of formylamide XIV according to the synthetic scheme shown in Scheme 2 is as illustrated in Examples 1-6.

Example 1 Acyloxazolidinone (IX) a. Acyloxazolidinone IXb (R ² = n-Pr; R ⁸ = CH ₂ Ph): 75 m
(S)-(-)-4-benzyl-2-oxazolidinone in anhydrous THF (4.0 g).
, 22.6 mmol) in cold (-78 ° C) solution over 15 minutes in hexane (9.
(1 mL, 22.6 mmol) in a 2.5 M solution of n-BuLi. After 5 minutes, pure valeryl chloride (2.95 mL, 24.9 mmol) was added dropwise and the mixture was-
Stirred at 78 ° C. for an additional 45 minutes. The mixture was then allowed to reach room temperature, stirred for a further 90 minutes, and then treated with 50 mL of a saturated NH ₄ Cl solution. Then, dichloromethane (50 mL) was added and the organic layer was washed with brine (2 × 30).
mL), dried over MgSO ₄ and concentrated in vacuo. This gave 5.94 g (100%) of the desired acyloxazolidinone I as a clear, colorless oil.
Xb was obtained. ¹ H NMR (300 MHz, CDCl ₃ ) δ7.36-7.20 (m, 5H), 4.7
1-4.64 (m, 1H), 4.23-4.14 (m, 1H), 3.40 (dd, J = 13
.3, 3.2 Hz, 1H), 3.04-2.84 (m, 2H), 2.77 (dd, J = 1 3.3, 9.6 Hz, 1H), 1.74-1.63 (m, 2H), 1.46-1.38 (m
, 2H), 0.96 (t, J = 7.3 Hz, 3H)

B. Acyl oxazolidinone ^{IXa (R 2 = Et; R} 8 = CH 2 Ph): by Ashiruo Kisazorijinon IXb analogous procedure to that described for the preparation of, (S) - (-) - Lithium anion of 4-benzyl-2-oxazolidinone Was treated with butyryl chloride to give the acyl oxazolidinone IXa in 94% yield. ¹ H NMR (300 MHz, CDCl ₃ ) δ7.37-7.20 (m, 5H), 4.68 (ddd, J = 13.1, 7.0, 3.4 Hz, 1H), 4.23-4 .13 (m,
2H), 3.30 (dd, J = 13.3, 9.6 Hz, 1H), 3.02-2.82 (m, 2H), 2.77 (dd, J = 13.3, 9.6 Hz) , 1H), 1.73 (q, J = 7
.3Hz, 2H), 1.01 (t, J = 7.3Hz, 3H)

C. Acyl oxazolidinone ^{IXc (R 2 = n-Bu} ; R 8 = CH 2 Ph): by a method analogous to that described for the preparation of reed oxazolidinone IXb, (S) - (- ) - 4- benzyl-2-oxazolidinone Was treated with hexanoyl chloride to give the acyloxazolidinone IXc in 96% yield. ¹ H NMR (300 MHz, CDCl ₃ ) δ7.36-7.20 (m, 5H), 4.68 (m, 1H), 4.23-4.14 (m, 2H), 3.30 (dd, J = 13.3, 3.3 Hz, 1H), 3.02-2.83 (m, 2H), 2.76 (dd, J = 13.3, 9.6 Hz, 1H), 1.70 (m , 2H), 1.43-1.34 (m, 4H), 0.9
2 (t, J = 3.3 Hz, 3H)

D. Acyl oxazolidinone ^{IXd (R 2 = i-Bu} ; R 8 = CH 2 Ph): i. 4- methylvaleryl chloride 4-methylvaleryl chloride, the following method was prepared from commercially available 4-methyl valeric acid; cooling of 4-methyl valeric anhydride in CH ₂ Cl ₂ 50mL containing DMF in 10 mL (0 ° C. ) Solution (1.85 mL, 15.0 mmol) was treated with 1.95 μL of oxalyl chloride (22.5 mmol). The mixture was then stirred at room temperature for 3 hours, concentrated in vacuo and filtered to give 1.65 g (100%) of the desired acid chloride as a colorless liquid. ii. Acyl oxazolidinone ^{IXd (R 2 = i-Bu} ; R 8 = CH 2 Ph): by a method analogous to that described for the preparation of acyl oxazolidinone IXb, (S) - (- ) - 4-benzyl-2-oxazolidinone Treatment of the lithium anion with 4-methylvaleryl chloride provided the acyloxazolidinone IXd in 85% yield. _{^{1 H NMR (300MHz, CDCl 3}} ) δ7.37-7.20 (m, 5H), 4. 70-4.63 (m, 1H), 4.23-4.15 (m, 2H), 3. 30 (dd, J = 13.2, 3.2 Hz, 1H), 2.98-2.90 (m, 2H), 2.76 (dd, J = 13.3, 9.6 Hz, 1H), 1 .68-1.54 (m, 3H), 0.94 (d, J = 6.2 Hz, 3H)

C. Acyloxazolidinone IXe (R ² ＣＨCH ₂ Ph; R ⁸ ＣＨCH ₂ Ph): By a procedure similar to that described for the preparation of acyl oxazolidinone IXb, (S)-(−)-4-benzyl-2-oxazolidinone The lithium anion is treated with hydrocinnamoyl chloride to give the acyloxazolidinone IX in 82% yield.
e was obtained. ¹ H NMR (300 MHz, CDCl ₃ ) δ7.35-7.16 (m, 10H), 4
.70-4.63 (m, 1H), 4.21-4.14 (m, 2H), 3.38-3.19 (m
, 3H), 3.08-2.98 (m, 2H), 2.75 (dd, J = 13.4, 9.5 Hz, 1H).

Example 2: Acyl oxazolidinones (X) a. Acyloxazolidinone Xb (R^Two= N-Pr; R⁸= CH_TwoPh): 110 mL of anhydrous CH_TwoCl_TwoA cooled (0 ° C.) solution of acyloxazolidinone IXb (5.74 g, 22.0 mmol) in 2.52 mL of TiCl_Four(23.1 mmol) to form a rich precipitate. After 5 minutes, diisopropylethylamine (4.2
(2 mL, 24.7 mmol) was added slowly and the resulting dark brown solution was allowed to stand at room temperature for 35
Stirred for minutes. Benzyl chloromethyl ether (6.0 mL, 44.0 mmol) was added.
Added quickly and the mixture was stirred at room temperature for 5 hours. Then 50 mL of CH _Two Cl_TwoAnd 75 mL of 10% NH_FourUpon addition of the aqueous Cl solution, a yellow gum was formed. After vigorously stirring the suspension for 10 minutes, the supernatant was placed in a separatory funnel.
Transfer and remove the gummy residue to 100 mL of 1: 1 10% NH_FourCl aqueous solution / CH_TwoCl_TwoTo
Collected. The combined organic layers were then combined with 1N aqueous HCl, saturated NaHCO 3_ThreeAnd
And brine, then wash with MgSO_FourDry over and concentrate in vacuo. The crude solid was recrystallized from EtOAc / hexane to give 6.80 g of the desired acyloxazolidinone Xb as a white solid in 81% yield.¹1 H NMR (300 MHz, CDCl_Three) δ7.34-7.18 (m, 10H), 4
.77-4.69 (m, 1H), 4.55 (s, 2H), 4.32-4.23 (m, 1H), 4.21-4.10 (m, 2H), 3.80 (t, J = 9.0 Hz, 1H), 3.65 (
dd, J = 9.0, 5.0 Hz, 1H), 3.23 (dd, J = 13.5, 3.3 Hz, 1H), 2.69 (dd, J = 13.5, 9.3 Hz, 1H), 1.74-1.64 (m, 1H), 1.54-1.44 (m, 1H), 1.40-1.28 (m, 2H), 0.9
1 (t, J = 7.3 Hz, 3H) LRMS (FAB) m / e 382 (M + H⁺)

B. Acyl oxazolidinone ^{Xa (R 2 = Et; R} 8 = CH 2 Ph): acyloxy Sazorijinon by analogous procedure to that described for the preparation of Xb, 80%
Acyloxazolidinone Xa was obtained in a yield of 1. ¹ H NMR (300 MHz, CDCl ₃ ) δ7.36-7.18 (m, 10H), 4
.55 (s, 2H), 4.21-4.11 (m, 3H), 3.81 (t, J = 9.0 Hz,
1H), 3.66 (dd, J = 9.0, 5.0 Hz, 1H), 3.23 (dd, J = 13
.5, 3.2 Hz, 1H), 2.70 (dd, J = 13.5, 9.3 Hz, 1H), 1.78-1.57 (m, 2H), 0.94 (t, J = 7.5Hz, 3H)

C. Acyl oxazolidinone Xc (R ² = n-Bu; R ⁸ = CH ₂ Ph): By a procedure similar to that described for the preparation of acyl oxazolidinone Xb, 9
Acyloxazolidinone Xc was obtained with a yield of 1%. ¹ H NMR (300 MHz, CDCl ₃ ) δ7.38-7.17 (m, 10H), 4
.72 (m, 1H), 4.54 (s, 2H), 4.27-4.10 (m, 2H), 3.79 (
t, J = 8.7 Hz, 1H), 3.65 (dd, J = 9.1, 5.0 Hz, 1H), 3.
23 (dd, J = 13.5, 3.3 Hz, 1H), 2.68 (dd, J = 13.5, 9.
3Hz, 1H), 1.75-1.68 (m, 1H), 1.31-1.26 (m, 4H), 0.87 (t, J = 6.8Hz, 3H)

D. Acyl oxazolidinone Xd (R ² = i-Bu; R ⁸ = CH ₂ Ph): By a procedure similar to that described for the preparation of acyl oxazolidinone Xb, 9
Acyloxazolidinone Xd was obtained with a yield of 8%. ¹ H NMR (300 MHz, CDCl ₃ ) δ7.38-7.17 (m, 10H), 4
.75-4.67 (m, 1H), 4.57 (d, J = 12.0 Hz, 1H), 4.51 (d
, J = 12.0 Hz, 1H), 4.41-4.36 (m, 1H), 4.20-4.09 (m, 2H), 3.74 (t, J = 9.0 Hz, 1H) , 3.65 (dd, J = 9.0,5
.1 Hz, 1H), 3.23 (dd, J = 13.5, 3.2 Hz, 1H), 2.63 (dd, J = 13.5, 9.5 Hz, 1H), 1.74-1 .52 (m, 2H), 1.35 (dd, J = 13.1, 6.1 Hz, 1H), 0.92 (d, J = 2.9 Hz, 3H), 0.90 (d, J = 2.9Hz, 3H)

E. Acyl oxazolidinone _{^{Xe (R 2 = CH 2 Ph}} ; R 8 = CH 2 Ph): by a method analogous to that described for the preparation of acyl oxazolidinone Xb, 8
Acyloxazolidinone Xe was obtained with a yield of 4%. ¹ H NMR (300 MHz, CDCl ₃ ) δ7.38-7.15 (m, 15H), 4.
62-4.50 (m, 4H), 4.03 (dd, J = 9.0, 2.7 Hz, 1H), 3.93-3.82 (m, 2H), 3.66 (dd, J = 9.2, 4.8 Hz, 1H), 3.19 (dd, J = 13.5, 3.2 Hz, 1H), 2.98 (dd, J = 13.4, 8.
2 Hz, 1 H), 2.88 (dd, J = 13.4, 7.3 Hz, 1 H), 2.68 (dd
, J = 13.5, 9.3 Hz, 1H)

Example 3: Carboxylic acid (XI) a. Carboxylic acid XIb (R ² = n-Pr): 6.6 in 320 mL of THF / H ₂ O
A cooled (0 ° C.) solution of 0 g (17.3 mmol) of acyloxazolidinone Xb was prepared by adding 6.95 mL of 35% aqueous H ₂ O ₂ and 20 mL of lithium hydroxide monohydrate (1.46 g) in H ₂ O. , 34.6 mmol). 0 ° C
Stirred for 16 hours at, then, first, Na ₂ SO ₃ in H ₂ O in 55 mL (10.5 g)
The solution was then carefully treated with a solution of NaHCO ₃ (4.35 g) in 100 mL of H ₂ O. The mixture was stirred at room temperature for 30 minutes and concentrated under vacuum to remove THF. The resulting aqueous mixture was then washed with CH ₂ Cl ₂ (4 × 75 mL),
Cooled to 0 ° C., acidified with 6N aqueous HCl and extracted with CH ₂ Cl ₂ (1 × 200 mL and 3 × 100 mL). The combined organic layers were then dried over MgSO ₄ and concentrated in vacuo to give 3.47 g (90%) of the desired acid XIb as a clear, colorless oil. ¹ H NMR (300 MHz, CDCl ₃ ) δ 7.38-7.26 (m, 5H), 4.55 (s, 2H), 3.67 (m, 1H), 3.57 (dd, J = 9. 2, 5.2 Hz, 1 H), 2.75 (m, 1 H), 1.72-1.31 (m, 4 H), 0.93 (t, J = 7)
.2Hz, 3H) LRMS (FAB) m / e 223 (M + H ')

B. Carboxylic acid XIa (R ² = Et): Preparation of acyloxazolidinone XIb An acyloxazolidinone XIa was obtained in 48% yield by a procedure similar to that described for the preparation of acyloxazolidinone XIb. ¹ H NMR (300 MHz, CDCl ₃ ) δ7.36-7.27 (m, 5H), 4.55 (s, 2H), 3.68 (dd, J = 9.2, 7.9 Hz, 1H), 3.59 (dd, J = 9.2, 5.4 Hz, 1H), 2.68-2.65 (m, 1H), 1.71-1.62 (m, 2H), 0.97 (t , J = 7.5Hz, 3H)

C. Carboxylic acid XIc (R ² = n-Bu): A procedure similar to that described for the preparation of the acyloxazolidinone XIb provided the acyloxazolidinone XIc in 96% yield. ¹ H NMR (300 MHz, CDCl ₃ ) δ7.37-7.28 (m, 5H), 4.55 (s, 2H), 3.67 (dd, J = 9.1, 8.1 Hz, 1H), 3.57 (dd, J = 9.2, 5.3 Hz, 1H), 2.72 (m, 1H), 1.67-1.51 (m, 2H), 1.36-1.27 (m , 4H), 0.89 (t, J = 6.9 Hz, 3H)

D. Carboxylic acid ^{XId (R 2 = i-Bu} ): by a method analogous to that described for the preparation of acyl oxazolidinone XIb, to give the acyl oxazolidinone XId in 80% yield. ¹ H NMR (300 MHz, CDCl ₃ ) δ7.37-7.28 (m, 5H), 4.55 (s, 2H), 3.64 (t, J = 9.1 Hz, 1H), 3.54 ( dd, J = 9.1, 5.1 Hz, 1H), 2.81 (m, 1H), 1.68-1.54 (m, 2H), 1.
36-1.27 (m, 1H), 0.92 (d, J = 4.9 Hz, 3H), 0.90 (d, J = 4.9 Hz, 3H)

E. Carboxylic acid XIe (R ² ＣＨCH ₂ Ph): A similar procedure to that described for the preparation of acyl oxazolidinone XIb afforded acyl oxazolidinone XIe in 92% yield. ¹ H NMR (300 MHz, CDCl ₃ ) δ 7.38-7.16 (m, 10H), 4
.53 (d, J = 12.1 Hz, 1H), 4.50 (d, J = 12.1 Hz, 1H), 3
.68-3.57 (m, 2H), 3.09-2.85 (m, 3H)

Example 4: Diethylamide (XII) a. Diethylamide XIIb (R ² = n-Pr; R ⁵ = R ⁶ = Et): diethylamine (2.36 mL, 23.0 mmol) and 2- (1H-benzotriazol-1-yl) -1,1,3 , 3-tetramethyluronium tetrafluoroborate (TBTU, 5.89 g, 18.4 mmol) 1 containing: 1 _{MeCN / CH 2 C l 2 (} 150mL) solution of the carboxylic acid XIb (3.40 g, 15.3 mmol) cooling
(0 ° C.) The solution was treated with diisopropylethylamine (6.7 mL, 38.2 mmol) for 1.5 hours (syringe pump). The mixture was then concentrated in vacuo and partitioned between ether (200 mL) and H ₂ O (100mL). The aqueous layer was further extracted with ether (2 × 100 mL) and the combined organic layers
Washed with aq. HCl (3 × 50 mL), aq. Sat. NaHCO ₃ and brine, dried over MgSO ₄ and concentrated in vacuo. Chromatographic purification (2
(30-400 mesh SiO ₂ , eluting with 1: 3 AcOEt / hexane) gave 4.24 g (97%) of diethylamide XIIb as a clear, colorless oil. ¹ H NMR (300 MHz, CDCl ₃ ) δ 7.35-7.23 (m, 5H), 4.52 (d, J = 12.0 Hz, 1H), 4.44 (d, J = 12.0 Hz, 1H) ), 3.
67 (t, J = 8.6 Hz, 1H), 3.51 (dd, J = 8.7, 5.5 Hz, 1H)
, 3.46-3.27 (m, 4H), 2.96 (m, 1H), 1.67-1.57 (m, 1
H), 1.48-1.22 (m, 4H), 1.20-1.10 (m, 6H), 0.90 (t, J = 7.2 Hz, 3H) LRMS (FAB) m / e 278 (M + H ⁺ )

B. Diethylamide ^{XIIa (R 2 = Et; R} 5 = R 6 = Et): diethylamide
A procedure similar to that described for the preparation of XIIb provided diethylamide XIIa in 73% yield. ¹ H NMR (300 MHz, CDCl ₃ ) δ7.33-7.26 (m, 5H), 4.52 (d, J = 12.0 Hz, 1H), 4.44 (d, J = 12.0 Hz, 1H) ), 3.
68 (t, J = 8.6 Hz, 1H), 3.53-3.33 (m, 5H), 2.90 (m, 1H), 1.75-1.50 (m, 2H), 1. 18 (t, J = 7.1 Hz, 3H), 1.
13 (t, J = 7.1 Hz, 3H), 0.89 (t, J = 7.4 Hz, 3H)

C. Diethylamide XIIc (R ² = n-Bu; R ⁵ = R ⁶ = Et): A procedure similar to that described for the preparation of diethylamide XIIb provided diethylamide XIIc in 94% yield. ¹ H NMR (300 MHz, CDCl ₃ ) δ 7.35-7.25 (m, 5H), 4.51 (d, J = 12.0 Hz, 1H), 4.44 (d, J = 12.0 Hz, 1H) ), 3.
67 (t, J = 8.6 Hz, 1H), 3.51 (dd, J = 8.8, 5.5 Hz, 1H), 3.46-3.29 (m, 1H), 2.94 (m , 1H), 1.66-1.62 (m, 2
H), 1.33-1.10 (m, 9H), 0.85 (t, J = 7.0 Hz, 3H)

D. Diethylamide ^{XIId (R 2 = i-Bu} ; R 5 = R 6 = Et): by Jiechirua bromide XIIb analogous procedure to that described for the preparation of, to obtain a diethylamide XIId in 95% yield. ¹ H NMR (300 MHz, CDCl ₃ ) δ 7.35-7.23 (m, 5H), 4.51 (d, J = 12.0 Hz, 1H), 4.44 (d, J = 12.0 Hz, 1H) ), 3.
65 (t, J = 8.7 Hz, 1H), 3.54-3.28 (m, 5H), 3.03 (m, 1H), 1.63-1.49 (m, 2H), 1. 33-1.24 (m, 1H), 1.18 (t, J = 7.1 Hz, 3H), 1.12 (t, J = 7.1 Hz, 3H), 0.90 (t,
J = 6.4Hz, 3H)

E. Diethylamide _{^{XIIe (R 2 = CH 2 Ph}} ; R 5 = R 6 = Et): by diethylamide XIIb analogous procedure to that described for the preparation of, to obtain a diethylamide XIIe in 89% yield. ¹ H NMR (300 MHz, CDCl ₃ ) δ7.35-7.16 (m, 10H), 4
.53 (d, J = 12.1 Hz, 1H), 4.47 (d, J = 12.1 Hz, 1H), 3
.77 (t, J = 8.5 Hz, 1H), 3.59 (dd, J = 8.8, 5.7 Hz, 1H)
), 3.40 (m, 1H), 3.22-2.89 (m, 5H), 2.79 (dd, J = 13
0.0, 5.1 Hz, 3H), 1.01 (t, J = 7.1 Hz, 3H), O.85 (t, J =
7.2Hz, 3H)

Example 5: Alcohol (XIII) a. Alcohol ^{XIIIb (R 2 = n-Pr} ; R 5 = R 6 = Et): 140mL of diethylamide XIIb (4.08 g, 14.7 mmol) in MeOH was added _{20% Pd (OH) 2 /} C (400mg of ) Was added and the suspension was hydrogenated at atmospheric pressure and room temperature for 15 hours. The catalyst was filtered and the filtrate was concentrated under vacuum to give 2.84 g (100%) of the desired primary alcohol XIIIb. ¹ H NMR (300 MHz, CDCl ₃ ) δ 3.74 (br.d, J = 4.2 Hz, 1 H), 3.61-3.15 (m, 5 H), 2.71 (m, 1 H), l. 69-1.24
(m, 4H), 1.20 (t, J = 7.1 Hz, 3H), 1.12 (t, J = 7.1 Hz, 3H), 0.92 (t, J = 7.2 Hz, 3H) LRMS (FAB) m / e 188 (M + H ⁺ )

B. Alcohol XIIIa (R ² = Et; R ⁵ = R ⁶ = Et): A procedure similar to that described for the preparation of alcohol XI IIb provided alcohol XIIIa in 100% yield. ¹ H NMR (300 MHz, CDCl ₃ ) δ 3.76 (m, 2H), 3.58-3.19 (m, 4H), 2.64 (m, 1H), 1.71-1.65 (m, 2H) 2H), 1.21 (t, J = 7.1 Hz, 3H), 1.13 (t, J = 7.1 Hz, 3H), 0.96 (t,
J = 7.4 Hz, 3H)

C. Alcohol ^{XIIIc (R 2 = n-Bu} ; R 5 = R 6 = Et): by a method analogous to that described for the preparation of alcohol XIIIb, to give the alcohol XIIIc in 100% yield. ¹ H NMR (300 MHz, CDCl ₃ ) δ 3.76 (d, J = 4.5 Hz, 2H)
3.58-3.19 (m, 4H), 2.72-2.65 (m, 2H), 1.68-1.5
5 (m, 2H), 1.40-1.24 (m, 4H), 1.20 (t, J = 7.1 Hz, 3H), 1.12 (t, J = 7.1 Hz, 3H) , 0.90 (t, J = 6.9 Hz, 3H)

D. Alcohol ^{XIIId (R 2 = i-Bu} ; R 5 = R 6 = Et): by a method analogous to that described for the preparation of alcohol XIIIb, to give the alcohol XIIId in 100% yield. ¹ H NMR (300 MHz, CDCl ₃ ) δ 3.78-3.68 (m, 2H), 3.57-3.15 (m, 4H), 2.81-2.73 (m, 1H), 1. 70-1.60 (m
, 2H), 1.40-1.28 (m, 1H), 1.21 (t, J = 7.1 Hz, 3H),
1.12 (t, J = 7.1 Hz, 3H), 0.92 (m, 6H)

E. Alcohol XIIIe (R ² ＣＨCH ₂ Ph; R ⁵ ＲR ⁶ ＥEt): alcohol
By a procedure similar to that described for the preparation of XIIIb, the alcohol XIIIe was obtained in 100% yield. _{^{1 H NMR (300MHz, CDCl 3}} ) δ7.29-7.16 (m, 5H), 3. 81-3.71 (m, 2H), 3.61-3.50 (m, 1H), 3. 15-2.87 (m
, 6H), 1.05 (t, J = 7.1 Hz, 3H), 0.98 (t, J = 7.1 Hz, 3H)
H)

Example 6: Aldehyde (XIV) a. Aldehyde ^{XIVb (R 2 = n-Pr} ; R 5 = R 6 = Et): prepared by wet CH ₂ Cl ₂ to (125 mL, and stirred CH ₂ Cl ₂ with water, separating the organic layer
To a solution of the alcohol XIIIb (2.34 g, 12.7 mmol) in Dess-Martin periodinane (8.06 g, 19.0 mmol). The mixture was stirred at room temperature for 40 minutes, then a 5% aqueous solution of Na ₂ S ₂ O ₃ containing 5.2 g of NaHCO ₃ (250 m
L) and a mixture of ether (200 mL). The biphasic mixture was stirred vigorously for 5 minutes and the aqueous layer was extracted with 15% CH ₂ Cl ₂ / Et ₂ O (2 × 100 mL). The combined organic layers were then washed with H ₂ O (3 × 75 mL) and brine, dried over MgSO ₄ , filtered, and concentrated in vacuo to give 2. as a clear, colorless oil.
06 g (88%) of the desired aldehyde XIVb were obtained. ¹ H NMR (300 MHz, CDCl ₃ ) δ 9.60 (d, J = 3.5 Hz, 1H)
, 3.49-3.30 (m, 5H), 1.96-1.85 (m, 2H), 1.39-1.3
1 (m, 2H), 1.19 (t, J = 7.1 Hz, 3H), 1.13 (t, J = 7.1 Hz, 3H), 0.95 (t, J = 7.3 Hz, 3H)

B. Aldehyde XIVb (R ² = n-Pr; R ⁵ = R ⁶ = Et): toluene (20 m
L), a solution of crude XIIIb (1.25 g, 6.68 mmol) in a mixture of ethyl acetate (20 mL) and water (3 mL) was added with free radical 2,2,6,6-tetramethyl-1. -Piperidinyloxy (TEMPO) (9 mg) was added. 0 ° C
Was cooled to, sodium hypochlorite solution prepared by adding sodium hypochlorite in 4.3mL into NaHCO ₃ in water 1.6g of 20 mL (10-13% available chlorine) for 30 minutes Over a small portion. Upon addition of sodium bromide (660 mg), the solution turned pale orange. Within minutes, the color of the reaction mixture turned grayish white. Additional sodium hypochlorite (4.7 mL) was added in several portions to complete the reaction. The aqueous layer was separated and extracted with toluene (20ml) and ethyl acetate (2x20mL). And the combined organic extracts were washed with a solution of KI (70 mg) in 10% aqueous KHSO ₄ solution. The organic layer was then washed with 5% Na ₂ S ₂ O ₃ and phosphate buffer at pH 7 and dried (
Na ₂ SO _4), to obtain a XIVb as concentrated to a pale yellow oily product (1.1 g). The spectral data for this compound was consistent with the product from Example 6a above.

C. Aldehyde ^{XIVa (R 2 = Et; R} 5 = R 6 = Et): by a method analogous to that described for the preparation of alcohol XIV b, to give the aldehyde XIVa in 80% yield. ¹ H NMR (300 MHz, CDCl ₃ ) δ 9.61 (d, J = 3.6 Hz, 1H)
3.48-3.29 (m, 5H), 2.02-1.90 (m, 2H), 1.19 (t, J = 7.1 Hz, 3H), 1.14 (t, J = 7.1 Hz, 3H), 0.96 (t, J = 7
.4Hz, 3H)

D. Aldehyde ^{XIVc (R 2 = n-Bu} ; R 5 = R 6 = Et): by a method analogous to that described for the preparation of alcohol X IVb, give the aldehyde XIVc in 98% yield. ¹ H NMR (300 MHz, CDCl ₃ ) δ 9.59 (d, J = 3.6 Hz, 1H)
, 3.48-3.29 (m, 5H), 1.97-1.87 (m, 2H), 1.39-1.2
2 (m, 4H), 1.18 (t, J = 7.2 Hz, 3H), 1.13 (t, J = 7.2 Hz, 3H), 0.90 (t, J = 7.0 Hz, 3H)

E. Aldehyde ^{XIVd (R 2 = i-Bu} ; R 5 = R 6 = Et): by a method analogous to that described for the preparation of alcohol X IVb, give the aldehyde XIVd in 96% yield. ¹ H NMR (300 MHz, CDCl ₃ ) δ 9.57 (d, J = 3.7 Hz, 1H)
3.51-3.27 (m, 5H), 1.83 (t, J = 7.1 Hz, 3H), l.66-1.55 (m, 1H), 1.20 (t, J = 7.1 Hz, 3H), 1.13 (t, J = 7.1 Hz, 3H), 0.93 (d, J = 6.6 Hz, 6H)

F. Aldehyde XIVe (R ² ＣＨCH ₂ Ph; R ⁵ ＲR ⁶ ＥEt): alcohol X
Aldehyde XIVe was obtained in 97% yield by a procedure similar to that described for the preparation of IVb. ¹ H NMR (300 MHz, CDCl ₃ ) δ 9.69 (d, J = 2.9 Hz, 1H)
, 7.29-7.16 (m, 5H), 3.65 (m, 1H), 3.53-3.42 (m, 1
H), 3.30 (dd, J = 13.5, 9.3 Hz, 1H), 3.23-3.13 (m, 2H), 3.06-2.91 (m, 2H), 1. 04 (t, J = 7.1 Hz, 3H), 0.
93 (t, J = 7.1 Hz, 3H)

The preparation of crust-lactacystin β-lactone and its analogs according to the synthetic reaction scheme is outlined in Reaction Scheme 1 as exemplified in Examples 7-9.

Example 7: Aldol adduct (II) a. Aldol adduct IIb (R^Two= N-Pr; R¹= I-Pr; R^Three= Me; R ^Four = Ph; R^Five= R⁶= Et): trans-oxazoline in ether (35 ml)
Ia (R¹= I-Pr; R^Four= Ph) in a cooled (-78 ° C) solution.
Tylsilyl) amide (1M solution in hexane, 2.17 mmol) was added. 30
After minutes, the orange solution was treated with dichloride in hexane (4.55 mL, 4.55 mmol).
Drop treatment with a 1 M solution of methylaluminum and cooling to −85 ° C. (liquid N_TwoTo
The mixture was stirred for an additional 60 minutes before being added to the dry ice / acetone bath).
Was. Then aldehyde XIVb (420 mg, 2.27 in ether (4 mL))
Mmol) was added along the wall of the flask over 10 minutes. The mixture was then allowed to warm to −40 ° C. for 2.5 hours and then 35 mL of saturated NH_FourQuenched by adding Cl and 25 mL of AcOEt. Then enough 2N HCl was added until two clear phases were obtained.
(About 15 mL added). The aqueous layer was extracted with AcOEt (2 × 20 mL), and the combined
The organic layer was washed with 0.5N HCl aqueous solution (20 mL), H_TwoO (20 mL), 0.5 M NaHSO_ThreeAqueous solution (2 × 15 mL), saturated NaHCO_ThreeAqueous solution and finally brine
, And then Na_TwoSO_FourAnd concentrated in vacuo.
879 mg (> 100%) of crude aldol produced, pure enough to be used
Compound IIb was obtained.¹1 H NMR (300 MHz, CDCl_Three) δ 8.02-7.97 and 7.53-7.39 (m, 5H), 6.58 (d, J = 9.9 Hz, 1H), 4.82 (d, J = 2.
4Hz, 1H), 3.73 (s, 3H), 3.69-3.61 (m, 2H), 3.49-3.39 (m, 2H), 3.24-3.16 (m, 1H), 3.05 (m, 1H), 2.89 (m, 1H), 2.28-2.23 (m, 1H), 1.98-1.91 (m, 1H), 1
.37-1.20 (m, 6H), 1.19-1.06 (m, 6H), 0.87 (t, J = 7.
1 Hz, 3H), 0.70 (d, J = 6.7 Hz, 3H) The aldol product IIb was also replaced with trans-oxazoline Ia.
The procedure is similar to that described above except that su-oxazoline Ib is used.
% Yield.

B. Aldol adduct IIb (R^Two= N-Pr; R¹= I-Pr; R^Three= Me; R ^Four = Ph; R^Five= R⁶= Et): trans-oxazoline I in THF (280 mL)
a (R¹= I-Pr, R^Four= Ph) (20.74 g) to a cooled (-78 ° C) solution was added lithium bis (trimethylsilyl) amide (12.4M in hexane, 92.4 mL) over a period of 75 minutes. After 30 minutes, the orange solution was dissolved in hexane (202 mL).
Treated dropwise with a 1 M solution of dimethylaluminum chloride at -85 ° C.
(Liquid N_TwoBefore addition to a dry ice / acetone bath) for an additional 40 minutes.
Stirred. Then a solution of aldehyde XIVb (19.43 g) in THF (50 mL) was added over 45 minutes. The mixture is then released for 40 minutes.
And warmed to -20 ° C over a period of 25 minutes.
The yellow reaction mixture was cooled again to -78 ° C and then 40 mL of saturated NH_FourQuenched by careful addition of aqueous Cl solution. The reaction mixture was added to 46
0 mL saturated NH_FourPoured slowly into Cl aqueous solution. AcOEt (500 mL) was added and, with good stirring, the reaction mixture was acidified with 6N HCl to give two clear
Phase is obtained. The aqueous layer was extracted with AcOEt (2 × 200 mL) and the combined organic layers
To H_TwoO (2 × 200 mL), saturated NaHCO_ThreeAqueous solution (2 x 200 mL) and bra
Washed sequentially in (2 × 300 mL). Organic extract_TwoSO_FourAnd MgSO _Four Dry above and concentrate in vacuo to give 41.55 g of crude aldol product IIb pure enough to use directly in the next step. The spectrum data for this compound was consistent with that of the product from Example 7a above.

C. Aldol adduct ^{IIb (R 2 = Et; R} 1 = i-Pr; R 3 = Me; R 4 = Ph; R 5 = R 6 = Et): analogous procedure to that described for the preparation of aldol adduct IIb Treated the lithium anion of trans-oxazoline Ia (R ¹ = i-Pr, R ⁴ = Ph) sequentially with dimethylaluminum chloride and aldehyde XI Va to give the aldol adduct IIa in 95% yield. . ¹ H NMR (300 MHz, CDCl ₃ ) δ 8.00-7.97 and 7.51-7.39 (m, 5H), 6.50 (d, J = 9.9 Hz, 1H), 4.80 (d , J = 2.
4Hz, 1H), 3.81-3.64 (m, 2H), 3.74 (s, 3H), 3.45 (m
, 2H), 3.19 (m, 2H), 2.93-2.84 (m, 2H), 2.24 (m, 1H)
), 1.89 (m, 1H), 1.73-1.64 (m, 4H), 1.29 (t, J = 7.2 Hz, 3H), 1.12 (d, J = 6. 9 Hz, 3 H), 1.07 (d, J = 7.2 Hz)
, 3H), 0.70 (d, J = 6.7 Hz, 3H)

D. Aldol adduct IIc (R ² = n-Bu; R ¹ = i-Pr; R ³ = Me; R ⁴ = Ph; R ⁵ = R ⁶ = Et): similar to that described for the preparation of aldol adduct IIb The lithium anion of trans-oxazoline Ia (R ¹ = i-Pr, R ⁴ = Ph) is sequentially treated with dimethylaluminum chloride and aldehyde XI Vc to give the aldol adduct IIc in 100% yield. Obtained. ¹ H NMR (300 MHz, CDCl ₃ ) δ 8.02-7.98 and 7.53-7.33 (m, 5H), 6.57 (d, J = 10.0 Hz, 1H), 4.81 (d , J = 2.3 Hz, 1H), 3.73 (s, 3H), 3.68-3.60 (m, 2H), 3.49
-3.17 (m, 2H), 3.00 (m, 1H), 2.90 (m, 1H), 1.98-1.87 (m, 2H), 1.38-0.83 (m , 16H), 0.70 (d, J = 6.7 Hz, 3H)

E. Aldol adduct IId (R ² = i-Bu; R ¹ = i-Pr; R ³ = Me; R ⁴ = Ph; R ⁵ = R ⁶ = Et): similar to that described for the preparation of aldol adduct IIb The lithium anion of trans-oxazoline Ia (R ¹ = i-Pr, R ⁴ = Ph) is sequentially treated with dimethylaluminum chloride and aldehyde XI Vd to give the aldol adduct IId in 100% yield. Obtained. ¹ H NMR (300 MHz, CDCl ₃ ) δ 8.01-7.80 and 7.55-7.20 (m, 5H), 4.87 (d, J = 2.3 Hz, 1H), 3.73 (s) , 3H),
3.69-3.58 (m, 2H), 3.51-3.32 (m, 2H), 2.98-2.87 (m, 1H), 2.33-2.24 (m, 2H) 1H), 2.12-2.02 (m, 1H), 1
.83 (t, J = 7.1 Hz, 1H), 1.35 (t, J = 7.1 Hz, 3H), 1.25-1.05 (m, 5H), 0.93 (d, J = 6.6 Hz, 3H), 0.89 (d, J = 6.5 Hz, 3H), 0.80 (d, J = 6.5 Hz, 3H), 0.69 (d, J = 6)
.7Hz, 3H)

F. Aldol adduct IIb (R ² = CH ₂ Ph; R ¹ = i-Pr; R ³ = Me; R ⁴ = Ph; R ⁵ = R ⁶ = Et): similar to that described for the preparation of aldol adduct IIb The lithium anion of trans-oxazoline Ia (R ¹ = i-Pr; R ⁴ = Ph) is sequentially treated with dimethylaluminum chloride and aldehyde XI Ve to give the aldol adduct IIe in 100% yield. Obtained. ¹ H NMR (300 MHz, CDCl ₃ ) δ 8.01-7.93 and 7.54-7.10 (m, 10H), 4.71 (d, J = 2.5 Hz, 1H), 3.73 (s) , 3H)
3.68-3.58 (m, 2H), 3.48-2.79 (m, 6H), 2.17 (m, 1
H), 1.12-0.91 (m, 9H), 0.68 (d, J = 6.7 Hz, 3H)

Example 8: γ-lactam (IV) a. γ-lactam IVb (R ² = n-Pr; R ¹ = i-Pr; R ³ = Me): 4.8
A solution of the aldol adduct IIb (4.72 g, 10.9 mmol) in 100 mL of 1: 9 AcOH / MeOH with addition of 20% Pd (OH) ₂ / C was added to 55 p.
.S.I 60 hours under H ₂ for. vigorously shaken. The mixture was cooled to ambient temperature before being filtered and concentrated under vacuum. The resulting solid was purified by flash chromatography (SiO ₂ , eluting with 1: 1 AcOEt / 1% AcOH in hexane) to afford 2.23 g (75%) of the desired product as a white solid. γ-lactam IVb was obtained. ¹ H NMR (300 MHz, CDCl ₃ ) δ 7.89 (br.s, 1H), 4.77 (br.d, J = 11.5 Hz, 1H), 4.47 (dd, J = 11.5,5) .6 Hz, 1H), 4.08 (dd, J = 9.4, 5.0 Hz, 1H), 3.83 (s, 3H), 2.
93 (m, 1H), 1.78-1.39 (m, 6H), 1.02-0.88 (m, 9H)

B. γ-lactam IVa (R ² = Et; R ¹ = i-Pr; R ³ = Me): The aldol adduct IIa was converted to 55 psi by a procedure similar to that described for the preparation of γ-lactam IVb. Hydrogenated under H _{2 for} 48 hours to give γ in 72% yield
-Lactam IVa was obtained. ¹ H NMR (300 MHz, CDCl ₃ ) δ 7.79 (br.s, 1 H), 4.62 (
br.d, J = 11.2 Hz, 1H), 4.51 (dd, J = 11.2, 5.4 Hz, 1H), 3.83 (s, 3H), 2.85 (m, 1H), 1.77-1.64 (m, 3H), 1.01 (t, J = 7.4 Hz, 3H), 0.98 (d, J = 6.9 Hz, 3H), 0
.95 (d, J = 6.9 Hz, 3H)

C. γ-lactam IVc (R ² = n-Bu; R ¹ = i-Pr; R ³ = Me): in 6 mL of 1: 9 AcOH / MeOH with addition of 250 mg of 20% Pd (OH) ₂ / C. Of an aldol adduct IIc (361 mg, 0.80 mmol) in 50 p.
Shake vigorously under si H _{2 for} 24 h. Then, a further catalyst (100 m
g) was added and the mixture was shaken again under 50 psi of H _{2 for a} further 24 h, then lowered to ambient temperature before filtration. The filtrate was then heated at reflux for 30 minutes, cooled to room temperature and concentrated in vacuo. The resulting solid was once evaporated with toluene and purified by flash chromatography (eluting with _{SiO 2, 4% Me OH /} CHCl 3), as a white solid 140 mg (61%
) To obtain the desired γ-lactam IVc. ¹ H NMR (300 MHz, CDCl ₃ ) δ 8.02 (br.s, 1H), 4.93 (
br.d, J = 11.3 Hz, 1 H), 4.46 (dd, J = 1 l 3, 5.5 Hz, 1
H), 4.15-4.08 (m, 1H), 3.83 (s, 3H), 2.94-2.87 (m, 1H), 1.80-1.34 (m, 6H) , 0.94 (d, J = 6.9 Hz, 3H), 0.89 (t, J = 7.2 Hz, 3H)

D. γ-lactam IVd (R ² = i-Bu; R ¹ = i-Pr; R ³ = Me): The aldol adduct IId was converted to 55 pps by a procedure similar to that described for the preparation of γ-lactam IVc. Hydrogenation under i. for 40 hours and heating under reflux for 30 minutes to obtain γ-lactam IVd in 61% yield. ¹ H NMR (300 MHz, CDCl ₃ ) δ 7.92 (br.s, 1H), 4.81 (
br.d, J = 11.5 Hz, 1H), 4.46 (m, 1H), 4.09 (m, 1H), 3.83 (s, 3H), 3.04-2.98 (m, 1H), 1.78-1.73 (m, 2H
), 1.66-1.47 (m, 3H), 1.00-0.90 (m, 12H)

E. γ-lactam IVe (R ² = CH ₂ Ph; R ¹ = i-Pr; R ³ = Me): γ-
By a procedure similar to that described for the preparation of lactam IVc, the aldol adduct IIe was hydrogenated at 50 psi for 24 hours and heated at reflux for 30 minutes to give γ-lactam IVe in 71% yield. I got ¹ H NMR (300 MHz, CDCl ₃ ) δ 8.01 (br.s, 1H), 7.35 −
7.15 (m, 5H), 5.02 (br.d, J = 11.7 Hz, 1H), 4.40-4.
34 (m, 1H), 4.06-4.01 (m, 1H), 3.84 (s, 3H), 3.34-
3.27 (m, 1H), 3.10-3.04 (m, 2H), 1.84-1.72 (m, 1H
), 0.98 (d, J = 6.7 Hz, 3H), 0.93 (d, J = 6.9 Hz, 3H)

Example 9: β-lactone (VII) a. β-lactone VIIb (R ² = n-Pr; R ¹ = i-Pr): To a cooled (0 ° C.) solution of γ-lactam IVb (2.20 g, 8.06 mmol) in EtOH (100 mL) , 0.1 N aqueous NaOH (100 mL, 10.0 mmol) were added. The mixture was stirred for 15 hours at room temperature, followed by the addition of H ₂ O (50mL) and AcOE t (100ml). The aqueous layer was then washed with AcOEt (2 × 50 mL), acidified with 6N aqueous HCl and concentrated in vacuo to a volume of about 60 mL. The solution was then frozen and lyophilized. The resulting solid was suspended in THF, filtered to remove sodium chloride and concentrated in vacuo to give 2.05 g (98%) of the desired dihydroxy acid as a white solid. ¹ H NMR (300 MHz, CD ₃ OD) δ 4.42 (d, J = 5.8 Hz, 1H)
, 3.90 (d, J = 6.5 Hz, 1H), 2.84 (m, 1H), 1.70-1.24 (
m, 6H), 0.95-0.84 (m, 9H) To a solution of dihydroxy acid (1.90 g, 7.33 mmol) in anhydrous THF (36 mL) was added 2- ( 1H-benzotriazol-1-yl)
-1,1,3,3-tetramethyluronium tetrafluoroborate (TBTU, 2
.59, 8.06 mmol) was added followed by triethylamine (0.72 mL, 22.0 mmol). After stirring at room temperature for 70 minutes, some toluene was added and the mixture was concentrated in vacuo and co-evaporated twice with toluene. Purification by flash chromatography (SiO ₂ , eluting with 2: 3 AcOEt / hexane) provided 1.44 g (81%) of the desired β-lactone VIIb as a white solid. ¹ H NMR (300 MHz, CDCl ₃ ) δ 6.07 (br.s, 1H), 5.26 (
d, J = 6.1 Hz, 1H), 3.97 (dd, J = 6.4, 4.4 Hz, 1H), 2.
70-2.63 (m, 1H), 2.03 (d, J = 6.4 Hz, 3H), 1.93-1.44 (m, 5H), 1.07 (d, J = 7.0 Hz) , 3H), 0.99 (d, J = 7.3 Hz, 3H), 0.91 (d, J = 6.7 Hz, 3H) LRMS (FAB) m / e 242 (M + H ⁺ )

B. β-lactone VIIa (R ² = Et; R ¹ = i-Pr): As described above for IVb, hydrolysis of IVa gave the corresponding dihydroxy acid in 100% yield. ¹ H NMR (300 MHz, CD ₃ OD) δ 4.45 (d, J = 5.8 Hz, 1H)
3.90 (d, J = 6.4 Hz, 1H), 2.74 (m, 1H), 1.71-1.53 (
m, 3H), 0.94 (t, J = 7.4 Hz, 3H), 0.92 (d, J = 6.8 Hz,
3H), 0.88 (d, J = 6.8 Hz, 3H) by a procedure similar to that described for the preparation of β-lactone VIIb.
Β-lactone VIIa was obtained in 9% yield. ¹ H NMR (300 MHz, CDCl ₃ ) δ 6.17 (br.s, ¹ H), 5.30 (
d, J = 6.0 Hz, 1H), 3.98 (dd, J = 6.4, 4.4 Hz, 1H), 2.
60 (m, 1H), 2.08 (d, J = 6.4 Hz, 3H), 1.97 (m, 2H), 1.
75 (m, 1H), 1.12 (t, J = 7.5 Hz, 3H), 1.07 (d, J = 6.8 Hz, 3H), 0.92 (d, J = 6.8 Hz, 3H)

C. β-lactone VIIc (R ² = n-Bu; R ¹ = i-Pr): As described above for IVb, hydrolysis of IVc gave the corresponding dihydroxy acid in 100% yield. ¹ H NMR (300 MHz, CD ₃ OD) δ 4.42 (d, J = 5.8 Hz, 1H
), 3.90 (d, J = 6.4 Hz, 1H), 2.86-2.79 (m, 1H), 1.70
-1.24 (m, 8H), 0.97-0.86 (m, 9H) by a procedure similar to that described for the preparation of β-lactone VIIb.
Β-lactone VIIc was obtained with a yield of 0%. ¹ H NMR (300 MHz, CDCl ₃ ) δ 6.14 (br.s, ¹ H), 5.27 (
d, J = 6.1 Hz, 1H), 3.97 (d, J = 4.4 Hz, 1H), 2.68-2.
61 (m, 1H), 1.94-1.86 (m, 2H), 1.72-1.36 (m, 7H),
1.07 (d, J = 7.0 Hz, 3H), 0.93 (t, J = 7.1 Hz, 3H), 0.91 (d, J = 6.8 Hz, 3H) LRMS (FAB) m / e 256 (M + H ⁺ )

D. β-lactone VIId (R ² = i-Bu; R ¹ = i-Pr): As described above for IVb, hydrolysis of IVd gave the corresponding dihydroxy acid in 100% yield. ¹ H NMR (300 MHz, CD ₃ OD) δ 4.50 (d, J = 5.8 Hz, 1H)
, 4.00 (d, J = 6.5 Hz, 1H), 3.09-3.02 (m, 1H), 1.90-1.61 (m, 3H), 1.49-1.40 ( m, 2H), 1.02 (d, J = 6.7H
z, 3H), 0.98 (d, J = 6.5 Hz, 3H), 0.97 (d, J = 6.7 Hz,
3H) By a procedure similar to that described for the preparation of β-lactone VIIb, 6
Β-lactone VIId was obtained in a yield of 2%. ¹ H NMR (300 MHz, CDCl ₃ ) δ 6.16 (br.s, ¹ H), 5.25 (
d, J = 6.1 Hz, 1H), 3.97 (d, J = 4.4 Hz, 1H), 2.71 (dd
, J = 15.1, 6.2 Hz, 1H), 1.95-1.66 (m, 5H), 1.08 (d, J = 6.9 Hz, 3H), 0.99 (d, J = 6.3 Hz, 3H), 0.98 (d, J
= 6.3 Hz, 3H), 0.92 (d, J = 6.7 Hz, 3H) LRMS (FAB) m / e 256 (M + H ⁺ )

E. β-lactone VIIe (R ² = CH ₂ Ph; R ¹ = i-Pr): For IVb, hydrolysis of IVe gave the corresponding dihydroxy acid in 88% yield, as described above. ¹ H NMR (300 MHz, CD ₃ OD) δ 7.25-7.04 (m, 5H), 4.29 (d, J = 5.7 Hz, 1H), 3.83 (d, J = 6.4 Hz, 1H), 3.01
-2.82 (m, 3H), 1.65 (m, 1H), 0.90 (d, J = 6.6 Hz, 3H)
, 0.86 (d, J = 6.8 Hz, 3H) by a procedure similar to that described for the preparation of β-lactone VIIb.
Β-lactone VIIe was obtained in a yield of 7%. ¹ H NMR (300 MHz, CDCl ₃ ) δ7.36-7.20 (m, 5H), 6.57 (br.s, 1H), 5.08 (d, J = 5.4 Hz, 1H), 3. 94 (d, J = 4.5 Hz, 1H), 3.25 (d, J = 10.1 Hz, 1H), 3.01-2.89 (m, 2H), 1.92-1.81 (m , 1H), 1.05 (d, J = 6.9 Hz, 3H), 0.86 (d, J = 6.7 Hz, 3H) LRMS (FAB) m / e 290 (M + H ⁺ )

The preparation of cis-oxazoline and trans-oxazoline according to the synthetic reaction scheme is shown in Reaction Schemes 3 and 4 as illustrated in Examples 10 and 11. Example 10: cis-oxazoline (Ia) a. Ethyl 3- (isopropyl) propionate (XV; R ¹ = i-Pr; R ³ = Me): Carbohydrate in dry CH ₂ Cl ₂ (168 mL) at 0 ° C. To a stirred solution of methoxymethylenetriphenylphosphorane (56.04 g, 167.6 mmol) was added isobutyraldehyde (17.4 mL, 191.6 mmol) dropwise. After 5 minutes, the reaction mixture was warmed to room temperature and stirred for 24 hours. The solvent was removed in vacuo and pentane was added to the white oily solid to precipitate triphenylphosphine oxide. The solid was filtered off and the filtrate was concentrated in vacuo. The procedure was repeated once more to give the crude olefin (20.00 g, 93%) as a yellow oil which was sufficiently pure for the next step. ¹ H NMR (300 MHz, CDCl ₃ ) δ 6.95 (dd, J = 15.7, 6.6)
Hz, 1H), 5.77 (dd, J = 15.7, 1.5 Hz), 3.72 (s, 3H),
2.44 (m, 1H), 1.06 (d, J = 6.7 Hz, 6H)

B. (2S, 3R) -methyl 2,3-dihydroxy-3- [isopropyl] propionate (XVIa; R ¹ = i-Pr; R ³ = Me): AD-mix-β (100.00.0 g), methanesulfonamide (6.78 g, 71.3 mmol) and a mixture of tert-butanol-water (1: 1, 720 mL) were stirred vigorously at room temperature for 5 minutes. The reaction mixture was then cooled to 0 ° C. and the α, β-unsaturated ester XV (R ¹ = i-Pr; R ³ = Me) (9.14 g, 71.3 mmol) was passed through a Pasteur pipette. It was dropped. After stirring at 0 ° C. for 96 hours, Na ₂ SO ₃ (3.0 g) was added, and stirring was continued at room temperature for 1 hour. The mixture was diluted with ethyl acetate (200ml) and transferred to a separatory funnel. Remove the organic layer and wash the aqueous phase with ethyl acetate (2 × 100 mL)
Extracted. The combined organic layers were dried (Na ₂ SO ₄ ), filtered and concentrated in vacuo. The resulting yellow oil was passed through a silica gel pad using 1: 1 hexane / ethyl acetate to afford the diol XVIa (R ¹ = i-Pr; R ³ = Me) as a yellow solid (11. (48 g, 94%). ¹ H NMR (300 MHz, CDCl ₃ ) δ 4.28 (dd, J = 5.6, 1.8 H
z, 1H), 3.80 (s, 3H), 3.48 (m, 1H), 3.28 (m, 1H), 2.33 (d, J = 9.3 Hz, 1H), 1.87 (m, 1H), 1.02 (d, J = 6.7 Hz, 3H), 0.95 (d, J = 6.7 Hz, 3H)

C. (2S, 3R) -methyl 2-bromo-3-dihydroxy-3- (isopropyl
) Propionate (XVIIa; R¹= I-Pr; R^Three= Me): (2S, 3R) -me
Cyl 2,3-hydroxy-3- [isopropyl] propionate XVIa (R¹ = I-Pr; R^Three= Me) (1.0 g, 6.17 mmol) and trimethylortho
The benzoic acid (1.02 mL, 80.1 mmol) was added to CH_TwoCl_Two(20 mL) and BF _Three ・ OEt_Two(40.0 μL, 0.32 mmol). After stirring for 75 minutes
The mixture was concentrated under full vacuum (0.05 mmHg) for 35 minutes. The mixture is CH_TwoCl_Two(20 mL), cooled to 0 ° C., and_ThreeN (43.0 μL, 0.31 mmol) and acetyl bromide (0.48 mL, 6.49 mmol) sequentially
Processed. After stirring at 0 ° C. for 4 hours, the reaction mixture was washed with saturated NaHCO 3_Threesolution(
12 mL) and allowed to warm to room temperature. The layers were separated and the aqueous layer was washed with CH_Two Cl_Two(2 × 20 mL). Dry the combined organic layers (Na_TwoSO_Four), Filtered and concentrated in vacuo to give crude α-bromo β-benzoic acid as a clear, colorless oil.
XVIIa (R¹= I-Pr; R^Three= Me) (1.36 g, 85%).¹1 H NMR (300 MHz, CDCl_Three) δ 8.05-8.00 (m, 2H), 7.47-7.40 (m, 3H), 5.57 (dd, J = 8.8, 3.9 Hz, 1H), 4.47 ( d, J = 8.8 Hz, 1H), 3.67 (s, 3H), 2.45 (m, 1H), 1.
01 (d, J = 6.8 Hz, 6H)

D. (2S, 3R) -methyl 2-azo-3-hydroxy-3- [isopropyl] propionate (XVIIIa; R ¹ = i-Pr; R ³ = Me): 15 mL of DMSO
(2R, 3R) -methyl 2-bromo-3-hydroxy-3- [isopropyl]
A solution of propionate XVIIa (R ¹ = i-Pr; R ³ = Me) (2.00 g, 6.07 mmol) was treated with sodium azide (790.0 mg, 12.2 mmol). After stirring for 12 h at room temperature, the mixture was partitioned H ₂ O and ethyl acetate (each 50ml) therebetween. Extracted with the aqueous layer was further ethyl acetate, dried the combined organic layers over MgSO _4, alpha-azo dried in vacuo as a yellow oily product
β-benzoic acid (1.55 g, 87%) was obtained. ¹ H NMR (300 MHz, CDCl ₃ ) δ 8.07-8.02 (m, 2H), 7.55-7.43 (m, 3H), 5.40 (dd, J = 8.8, 2.8 Hz) , 1H), 3.73 (s, 3H), 2.24 (m, 1H), 1.04 (d, J = 5.8 Hz, 3H), 0.
98 (d, J = 5.8 Hz, 3H) The same procedure was repeated except that DMF was used instead of DMSO as the solvent.
The desired α-azo β-benzoic acid was obtained with a yield of 85%.

E. Benzamide XIXa (R ¹ = i-Pr; R ³ = Me): ethyl acetate (25 m
(2S, 3R) -Methyl 2-azo-3-dihydroxy-3- [isopropyl] propionate XVIIIa (R ¹ = i-Pr; R ³ = Me) (1.50 g, 5.
(15 mmol) was treated with 200 mg of 20% Pd (OH) ₂ / C and the suspension was stirred vigorously in a H ₂ atmosphere under balloon pressure. After 12 hours, the mixture was filtered and refluxed for 4 hours to complete the transfer of the benzoyl group. The mixture was then cooled to room temperature and concentrated in vacuo to give the desired benzamide (1.25 g, 92%) as a yellow oil. ¹ H NMR (300 MHz, CDCl ₃ ) δ 7.85-7.83 (m, 2H), 7.46-7.40 (m, 3H), 6.99 (br.d, J = 9.1 Hz, 1H ), 5.05
(dd., J = 9.1, 1.9 Hz, 1H), 3.77 (s, 3H), 1.79 (m, 1H), 1.03 (d, J = 6.7 Hz, 3H) 0.99 (d, J = 6.7 Hz, 3H)

F. Cis-oxazoline Ia (R ¹ = i-Pr; R ³ = Me): CH ₂ Cl ₂ (20 m
L) in 500mg benzamide ^{XIXa (R 1 = i-Pr} ;. A solution of R ³ = Me) (18 8 mmol) was treated with 4.50mL of thionyl chloride (61.7 mmol). After stirring at room temperature for 24 hours, the mixture was diluted with CH ₂ Cl ₂ and saturated Na 2
Wash with HCO ₃ solution, dry (Na ₂ SO ₄ ), concentrate in vacuo, chromatograph (silica gel, 1: 1 hexane / ethyl acetate) to give the desired cis-oxazoline (248 mg). , 53%) as a pale yellow oil. ¹ H NMR (300 MHz, CDCl ₃ ) δ 8.01-7.97 (m, 2H), 7.
52-7.38 (m, 3H), 4.94 (d, J = 9.8 Hz, 1H), 4.53 (dd, J = 9.8, 7.8 Hz, 1H), 3.76 (s , 3H), 2.09 (m, 1H), 1.
05 (d, J = 6.5 Hz, 3H), 1.01 (d, J = 6.7 Hz, 3H)

Example 11: trans-oxazoline (Ib) a. Ethyl 3- (isopropyl) propionate (XV; R ¹ = i-Pr; R ³ = Me): dry CH ₂ Cl ₂ (168 mL) at 0 ° C. To a stirred solution of carbomethoxymethylene triphenylphosphorane (56.04 g, 167.6 mmol) in was added isobutyraldehyde (17.4 mL, 191.6 mmol) dropwise. After 5 minutes, the reaction mixture was warmed to room temperature and stirred for 24 hours. The solvent was removed in vacuo and pentane was added to the white oily solid to precipitate triphenylphosphine oxide. The solid was filtered off and the filtrate was concentrated in vacuo. This procedure was repeated one more time to give the crude olefin (20.00 g, 93%) as a yellow oil that was sufficiently pure for the next step. ¹ H NMR (300 MHz, CDCl ₃ ) δ 6.95 (dd, J = 15.7, 6.6)
Hz, 1H), 5.77 (dd, J = 15.7, 1.5 Hz), 3.72 (s, 3H), 2.44 (m, 1H), 1.06 (d, J = 6. 7Hz, 6H)

B. (2R, 3S) -Methyl 2,3-hydroxy-3- [isopropyl] propionate (XVIb; R ¹ = i-Pr; R ³ = Me): K ₂ OsO ₂ (OH) ₄ (246.1m
g, 0.67 mmol, 0.95 mol%), hydroquinine, 1,4-phthalazinediyl diether (555.1 mg, 0.71 mmol, 1.01 mol%), N-methylmorpholine N-oxide (50 wt. %, 25.0 mL, 0.106 mol, 1.51 eq), t-BuOH (84mL) , and the clear yellow solution of H ₂ O (58mL), for 48 hours at 25 ° C., via a syringe pump Pure olefin XV (R ¹ = i-Pr; R ³ = Me) (9.0 g, 70.2 mmol) was added (a syringe was connected to the tube and the tip was immersed in the solution during the reaction time) ). Then, the resulting clear orange solution was stirred for an additional 60 minutes, then a solution of ethyl acetate (200 meters L) and _{H 2 O (150mL) Na 2} SO 3 in (15.0 g), obtained The resulting mixture was stirred for 4 hours. Separate the phases and separate the aqueous layer with more ethyl acetate.
(2x). Then, the organic layers were combined, extracted from the organic phase with a solution of chiral ligand of saturated _{Na 2 SO 4 (2 × 100mL} ) of 0.3M in H ₂ SO _4. The phases were separated once more and the aqueous layer was further extracted with ethyl acetate (1 ×). The organic layers were combined, dried over Na ₂ SO _4, filtered, and concentrated in vacuo. As a result, 11
0.4 g (about 100%) of a white oily solid was obtained, which consisted of 70% ee (diol in C ₆ D ₆ and tris [3- (heptafluoropropylhydroxymethylene)-(−)-europium camphorate). Determined by ¹ H NMR from a 1: 1 molar solution of
). 6.8 g by recrystallization from petroleum ether at 35-60 ° C.
(60%) of (2R, 3S) -methyl 2,3-dihydroxy-3- [isopropyl] propionate (XVIb; R ¹ = i-Pr; R ³ = Me), which is about 100% e
e, obtained as white crystals. mp = 32-34 ° C.; =-110.6 ° (c 1.04, CHCl ₃ )] ¹ H NMR (300 MHz, CDCl ₃ ) δ 4.28 (dd, J = 5.6, 1.8 H)
z, 1H), 3.80 (s, 3H), 3.48 (m, 1H), 3.28 (m, 1H), 2.33 (d, J = 9.3 Hz, 1H), 1.87 (m, 1H), 1.02 (d, J = 6.7 Hz, 3H), 0.95 (d, J = 6.7 Hz, 3H)

C. (2S, 3S) -Methyl 2-bromo-3-dihydroxy-3- (isopropyl)
Propionate (XVIIb; R ¹ = i-Pr; R ³ = Me): (2R, 3S) -methyl 2,3-dihydroxy-3- [isopropyl] propionate (XVIb; R ¹ = i-Pr; R ³ = Me) ) (30.0 g, 185.2 mmol) and trimethylorthobenzoic acid (41.3 mL, 240.7 mmol) were dissolved in CH ₂ Cl ₂ (400 mL) and BF ₃ OEt ₂ (1.16 mL, 9 .25 mmol). After 2 hours, triethylamine (1.8 mL, 13 mmol) was added and the mixture was concentrated in vacuo and placed under full vacuum (0.05 mmHg) for 70 minutes. The residue was re-dissolved in CH ₂ Cl ₂ (400 mL), cooled to 0 ° C. and acetyl bromide (14.3).
mL, 194.5 mmol). After 2 hours, additional acetyl bromide (0.
(68 mL, 9.25 mmol). After 30 minutes, a saturated NaHCO ₃ solution (50
0 mL) was added and the mixture was stirred vigorously for 5-10 minutes. Layers were separated and the aqueous layer was extracted with _{CH 2 Cl 2 (2 × 20mL} ). Dry the combined organic layers
(Na ₂ SO ₄ ), filtered and concentrated in vacuo to about 9% as a clear, colorless oil.
Crude α-bromo β-benzoic acid XVI Ib (R ¹ = i-Pr; R ³ = Me) containing 0.3% by weight of methyl benzoate (66.23 g) was obtained. For the product: ¹ H NMR (300 MHz, CDCl ₃ ) δ 8.05-8.00 (m, 2H), 7.47-7.40 (m, 3H), 5.57 (dd, J = 8.8). , 3.9 Hz, 1H), 4.47 (d, J = 8.8 Hz, 1H), 3.67 (s, 3H), 2.45 (m, 1H), 1.
01 (d, J = 6.8 Hz, 6H)

D. (2R, 3S) -Methyl 2-azo-3-dihydroxy-3- [isopropyl] propionate (XVIIIb; R ¹ = i-Pr; R ³ = Me): sodium azide (24 g, 370 mmol) ) Was added to 230 mL of DMSO and the mixture was stirred at room temperature overnight. To the resulting solution was added (2S, 3S) -methyl 2-bromo-3-dihydroxy-3- [isopropyl] propionate (X in 20 mL of DMSO).
VIIb; was added a solution of ^{R 3 = Me) (61g,} 185 ^{mmol); R 1 = i-Pr} . After stirring at room temperature for 11 hours, the mixture was washed with water (1.5 L) and ether (
(200 mL) and stirred vigorously for 10-15 minutes. Ether (100 mL) was added and the layers were separated. The aqueous layer was extracted with ether (2 × 100 mL) and the combined organic layers were washed with water (2 × 100 mL) and brine (100 mL),
Dried over O ₄ and concentrated in vacuo to give a crude product (57.5 g) containing about 3% starting material and 8% disappeared by-products. For the product: ¹ H NMR (300 MHz, CDCl ₃ ) δ 8.07-8.02 (m, 2H), 7.55-7.43 (m, 3H), 5.40 (dd, J = 8.8). , 2.8 Hz, 1H), 3.73 (s, 3H), 2.24 (m, 1H), 1.04 (d, J = 5.8 Hz, 3H), 0.
98 (d, J = 5.8 Hz, 3H)

E. Benzamide XIXb (R ¹ = i-Pr; R ³ = Me): methanol (300
(2R, 3S) -Methyl 2-azo-3-dihydroxy-3- [isopropyl] propionate XVIIIb (R ¹ = i-Pr; R ³ = Me) (55 g) (0-5 ° C.) ) Add 94 mL of 4 M HCl / dioxane and 2.75 g of P
d (OH) ₂ / C was added. The mixture was purged with hydrogen and stirred at room temperature. The mixture was purged with hydrogen every 30 minutes to remove liberated nitrogen. After 4 hours, the reaction mixture was purged with nitrogen and additional Pd (OH) _{2 /} C (1.3 g) was added. The reaction mixture was purged with hydrogen and again purged every hour for 4 hours. The mixture was filtered and concentrated in vacuo. The residue was dissolved in water and extracted with EtOAc. The aqueous layer was basified with Na ₂ CO ₃ and extracted again with EtOAc. Wash the combined organic extracts with brine, dried over Na ₂ SO _4, and concentrated to give a mixture of directly using N- and O- benzoylated product in the next step.

F. Trans-oxazoline Ib (R ¹ = i-Pr; R ³ = Me): The crude product IXb (37.3 g, 141 mmol) obtained in Example 10e above was dissolved in toluene (350 mL). p-Toluenesulfonic acid (2.68 g, 14.1 mmol) was added and the mixture was heated to reflux. Transfer water to Dean Stark trap (D
ean Stark trap). After 3 hours, 〜2.5 mL of water was collected. The reaction mixture was cooled, diluted with EtOAc (100 mL), washed sequentially with saturated NaHCO ₃ (2 × 100 mL) and brine (100 mL), dried over MgSO ₄ and concentrated. The residue was purified on a pad of silica gel (〜400 g) eluting with 25-30% EtOAc-hexane to give trans-oxazoline Ib (R ¹ = i-Pr; R ³ = Me). ¹ H NMR (300 MHz, CDCl ₃ ) δ 8.01-7.97 (m, 2H), 7.52-7.38 (m, 3H), 4.68 (apparent t, J = 7 Hz, 1H), 4 .57 (d
, J = 7 Hz, 1H), 3.81 (s, 3H), 2.00-1.93 (m, 1H), 1.04 (d, J = 6.7 Hz, 3H), 1.00 (d , J = 6.8Hz, 3H)

Example 12 Inactivation of Proteasome Activity Purification of the 20S proteasome and proteasome activator PA28 was performed as previously described (Dick et al., J. Biol. Chem. 271: 7273 (1996)). ). 2 mL of assay buffer (20 mM HEPES, 0.5 mM EDTA, pH
8.0) and Suc-Leu-Leu-Val-Tyr-AMC in dimethyl sulfoxide were added to a 3 mL fluorescent cuvette and the cuvette was replaced with a Hitachi F-2.
000 fluorescence spectrometer into a jacketed cell holder. The temperature was maintained at 37 ° C. by a circulating water bath. 0.34 mg of PA28 was added and the progress of the reaction was monitored by the increase in fluorescence at 440 nm (λ _ex = 380 nm) associated with the formation of free AMC. The progress curve showed a lag phase lasting 1-2 minutes caused by the slow formation of the 20S-PA28 complex. After achieving a steady state of substrate hydrolysis, lactacystin was added to a final concentration of 1 mM and the reaction was monitored for 1 hour. fluorescence(
F) vs. time (t) data were collected on a microcomputer using LAB CALC (Galactic) software. The κ _inact value is a linear equation:

[0152]

(Equation 1) Was inferred by a non-linear least squares fit of the data to

Example 13 Inhibition of Intracellular Proteolysis in C2C12 Cells C2C12 cells (mouse myogenic cell line) were labeled with ³⁵ S-methionine for 48 hours. The cells were then washed and pre-incubated for 2 hours in the same medium supplemented with 2 mM unlabeled methionine. The medium was removed and replaced with a new aliquot of preincubation medium containing 50% serum and a concentration of the compound to be tested. Then, the medium was removed, finished to 10% TCA, and centrifuged. TCA soluble radioactivity was counted. Inhibition of protein hydrolysis is based on TCA
Calculated as percent reduction in soluble radioactivity. From this data, the IC _{50 of} each compound was calculated.

Example 14 Lactone Hydrolysis The half-life (t _1/2 ) for hydrolysis of the β-lactone analog to the corresponding dihydroxy acid was determined by comparing the 200 mM HEPES, pH 7.8, 200 mM in 0.5 mM EDTA. The concentration was measured at 37 ° C. The absorbance was measured at 230 nm where there was the largest difference in the absorbance coefficient between lactone and dihydroxy at that wavelength for at least 5 half-lives (approximately 1 hour). Half-life was determined by Guggenheim analysis (Enzymes by Gutfred und: Physical Principles; Wi).
Ley and Sons: New York, 1975, pp 118-119.
). The results of Examples 12-14 are reported in Table 1.

[0155]

Embedded image Table 1 Kinetics of inhibition of 20S proteasome and inhibition of intracellular proteolysis

[Table 1] ^a Inactivation of chymotrypsin-like activity of PA28-activated 20S proteasome ^b Inhibition of intracellular proteolysis in C2C12 cells ^c Half-life of hydrolysis Results indicate that compounds of the invention are potent inhibitors of proteasome .

Example 15 Reduction of Infarct Size and Neuronal Loss Methods Male Sprague Dawley rats (250-400 g) were anesthetized with halothane,
Central cerebral artery (MCA) occlusion was performed for 2 hours using nylon filaments. Subsequently, the filaments were removed and reperfusion of the infarcted tissue was performed 24 hours before the rats were killed.

Immediately after removing the filament, the animals were evaluated using a neurological scoring system. Neurological scores are expressed on a scale of 0 to 10, where 0 indicates no neurological deficits and 10 indicates severe neurological deficits. After 24 hours and before sacrifice, the animals were evaluated a second time using the same neurological scoring system. Staining of coronal sections (2.0 mm x 7-8) with triphenyltetrazolium chloride (TTC) taken throughout the brain was evaluated under blind conditions using image analysis to determine infarct size.

Dosing Mode Rats were treated with vehicle (50% propylene glycol / saline; n = 8) or 7-n-propyl-crust-lactacystin β-lactone (3b) 2 hours after the onset of occlusion. An iv bolus injection (1.0 mL / kg) of either 0.3 mg / kg: n = 7) was administered. Two additional groups of rats received an iv bolus injection of 3b (1.0 mL / kg) at 0 minutes, 2 hours and 6 hours after the onset of occlusion. One group (0.1 mg / kg: n = 6) received 0 mg at each of these times.
.1 mg / kg, while another group (0.3 mg / kg x 3: n = 7) received 0.3 mg / kg at each of the three time points.

Results In animals treated with a single dose of 7-n-propyl-crust-lactacysteine β-lactone (3b), the occluded volume was reduced by 50% (FIG. 1, 0.3 × 1).
). The infarct volume was not significantly reduced in either the 0.1 mg / kg x 3 administration group or the 0.3 mg / kg x 3 administration group (Fig. 1).

All animals had a neurological score of 10 ± 0 immediately after the 2 hour ischemic episode. At 24 hours, vehicle treated rats had an average score of 8.7 ± 0.6, but a single 0.3 mg / kg dose of 7-n-propyl-crust-lactacystin β-lactone ( Rats treated with 3b) had an average score of 4 ± 1 (FIG. 2). These data represent a 60% neurological improvement for drug treated animals. Neither the 0.1 mg / kg × 3 administration group nor the 0.3 mg / kg × 3 administration group showed any significant improvement in neurological score (FIG. 2).
.

Conclusions 7-n-Propyl-crust-lactacystin β-lactone given once after ischemia provides significant protection in both the extent of neurological deficits and infarcted brain injury. From these preliminary data, it appears that a single dose format is preferred over a multiple dose format.

Although the present invention has now been fully described, those skilled in the art will appreciate that a wide and equivalent range of conditions, prescriptions, and other It will be appreciated that the invention may be practiced within parameters. All patents and cited publications herein are expressly incorporated by reference.

[Brief description of the drawings]

BRIEF DESCRIPTION OF DRAWINGS FIG. 1 shows i versus infarct volume in rats (n = 6-8).
. v. It is a graph which shows the effect of the administered compound 3b.

FIG. 2 shows i.v. versus immunity score in rats (n = 6-8). v. It is a graph which shows the effect of the administered compound 3b.

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI Theme coat ゛ (Reference) A61P 29/00 A61P 29/00 35/00 35/00 35/04 35/04 43/00 111 43/00 111 C07C 231/10 C07C 231/10 233/82 233/82 235/80 235/80 C07D 263/16 C07D 263/16 491/044 491/044 // C07B 61/00 300 C07B 61/00 300 C07M 7: 00 C07M 7:00 (31) Priority claim number 60 / 067,352 (32) Priority date December 3, 1997 (Dec. 23, 1997) (33) Priority claim country United States (US) (81) Designated countries EP (AT, BE, CH, CY, DE, DK, ES, FI, FR, GB, GR, IE, IT, LU, MC, NL, PT, SE), OA (BF, BJ, CF, CG, CI, CM, GA, GN, GW, L, MR, NE, SN, TD, TG), AP (GH, GM, KE, LS, MW, SD, SZ, UG, ZW), EA (AM, AZ, BY, KG, KZ, MD, RU) , TJ, TM), AL, AM, AT, AU, AZ, BA, BB, BG, BR, BY, CA, CH, CN, CU, CZ, DE, DK, EE, ES, FI, GB, GE , GH, GM, HU, ID, IL, IS, JP, KE, KG, KP, KR, KZ, LC, LK, LR, LS, LT, LU, LV, MD, MG, MK, MN, MW, MX, NO, NZ, PL, PT, RO, RU, SD, SE, SG, SI, SK, SL, TJ, TM, TR, TT, UA, UG, UZ, VN, YU, ZW (71) Application People William Rouche 3105 West Dobson, Ann Arbor, Michigan 48105 USA (72) Inventor François Sucey United States 02174 Summit Street 60, Arlington, Mass. 72 (72) Inventor Louis Prammondon United States 02472-4922 Robbins Road 144, Watertown, Mass. 72 (72) Inventor Mark・ Banke, United States 02144, North Street 122, Somerville, Mass., Massachusetts, USA Inventor William Rouche, United States 48105, Ann Arbor, Michigan, West Dobson, 3116 F term (reference) 4C056 AA01 AB01 AC02 AD01 AE02 AF01 BA03 BA04 BB14 BC01 4C069 AB14 BC12 BD03 CC02 4C086 AA01 AA02 AA03 AA04 BC06 BC69 CB22 MA01 MA04 NA14 ZA36 ZA59 ZA66 ZB11 ZB13 ZB26 ZC20 4H006 AA01 AA02 AA03 AB22 AB23 AB29 AC11 AC28 AC41 AC44 AC48 AC52 AC53 AC59 AC81 BA25 BA17 BA66

Claims (73)

[Claims] (A) Formula I: embedded image Wherein R ¹ is alkyl, alkenyl, alkynyl, cycloalkyl, aryl, alkaryl, or aralkyl (wherein any of the aryl, aralkyl, or alkaryl ring moieties may be optionally substituted) R ³ is alkyl, alkenyl, alkynyl, cycloalkyl, aryl, or alkaryl, wherein any of these may be optionally substituted; R ⁴ is aryl or heteroaryl Wherein any of these may be optionally substituted)]] is treated with a strong base to deprotonate the substituted aryl or heteroaryl oxazoline to form an enolate; (B) the enolate is titanium, aluminum Transmetallized with a metal selected from the group consisting of iron, tin, zinc, magnesium and boron,
V: Wherein R ² is alkyl, cycloalkyl, aryl, alkaryl, aralkyl,
R ⁵ and R ⁶ are independently alkyl, hydroxy, alkoxyalkyl, or amide, wherein any of the aryl, aralkyl, or alkaryl ring moieties is optionally substituted; Or is one of alkaryl; or R ⁵ and R ⁶ , when taken together with the nitrogen atom to which they are attached, are optionally substituted and optionally further oxygen or nitrogen atoms To form a 5- to 7-membered heterocyclic ring, which may contain a compound of formula II: Wherein R ¹ to R ⁶ are as defined above; (c) catalytically hydrogenating the adduct of formula II to form an adduct of formula IV: Wherein R ¹ , R ² and R ³ are the same as defined above; and (d) saponifying the γ-lactam of formula IV to form a compound of the formula V: [Wherein R ¹ and R ² are the same as defined above], a method for forming a γ-lactam carboxylic acid represented by the formula V, or a salt thereof, comprising forming a carboxylic acid represented by the formula: 2. The method of claim 1, further comprising treating the carboxylic acid of formula V with a cyclizing reagent.
Formula VII: Wherein R ¹ and R ² are the same as defined in claim 1, wherein clast-lactacystin β-lactone is formed. 3. The method according to claim 1, wherein the cyclization is performed by arylsulfonyl chloride, benzotriazole-
1-yloxytris (dimethylamino) phosphonium hexafluorophosphate, O- (1H-benzotriazol-1-yl) -N, N, N ', N'-
3. The process according to claim 2, wherein the reaction is carried out with a reagent selected from the group consisting of tetramethyluronium tetrafluoroborate and alkyl-, aryl- or alkenyl-chloroformate. 4. The method of claim 1, further comprising a crust-lactacystin β of formula VII
Reacting the lactone with a thiol, R ⁷ SH, to form a compound of formula VI: Wherein R ¹ and R ² are the same as defined in claim 1; R ⁷ is alkyl, aryl, aralkyl or alkaryl (wherein any of the alkyl, aryl, aralkyl or alkaryl Optionally substituted)). 5. The method according to claim 4, wherein clast-lactacystin β-lactone is converted to lactacystin by treating β-lactone with N-acetylcysteine. 6. A carboxylic acid intermediate of formula V is coupled directly to a thiol, R ⁷ SH, to form a compound of formula VI: Wherein R ¹ and R ² are the same as defined in claim 1; R ⁷ is alkyl, aryl, aralkyl, or alkaryl (wherein each of the alkyl, aryl, aralkyl or alkaryl is Lactacystin having the formula (I) which may be substituted if desired). 7. In the step (a), the strong base is hindered.
2. The process according to claim 1, wherein the process is selected from the group consisting of :) amide bases; alkali metal hexamethyldisilazide; and constrained alkyllithium reagents. 8. The process according to claim 1, wherein in the step (a), the reaction is carried out in an ethereal solvent at a low temperature. 9. In step (a), the ethereal solvent is selected from the group consisting of diethyl ether, tetrahydrofuran, and dimethoxyethane, and the reaction temperature is from about -100 ° C to about -30 ° C. 9. The method according to claim 8, wherein 10. The process according to claim 1, wherein, in step (b), the enolate is transmetallated with titanium or aluminum or a mixture thereof. 11. The process according to claim 1, wherein in step (b), the enolate is transmetallated by reaction with Me ₂ AlCl. 12. The method according to claim 10, wherein 1 to 3 molar equivalents of said metal are used. 13. In step (c), the catalytic hydrogenolysis of adduct II gives aminodiol III: Wherein R ¹ -R ⁶ are the same as defined in claim 1 to obtain the desired γ-lactam (IV) as a mixture. 14. The hydrocracking in the presence of a catalyst selected from the group consisting of palladium black, palladium on activated carbon, and palladium hydroxide on carbon;
The method according to claim 1, wherein the reaction is performed in the presence of an organic solvent selected from the group consisting of lower alkanols, lower alkanoates, lower alkanoic acids, and mixtures thereof.
3. The production method according to 3. 15. The method according to claim 14, wherein the organic solvent is selected from the group consisting of methanol, ethanol, isopropanol, ethyl acetate, acetic acid, and a mixture thereof. 16. The process according to claim 13, wherein the crude product mixture is heated to convert aminodiol III to γ-lactam IV. 17. R¹But C_1-12Alkyl, C_3-8Cycloalkyl, C_2-8Alkenyl, C_2-8Alkynyl, C_6-14Aryl, C_6-10Ara (C_1-6) Lucil
Or C_1-6Arca (C_6-10) Is ril; R^TwoBut C_1-8Alkyl, C_3-8Cycloalkyl, C_2-8Alkenyl, C_2-8Alkynyl, C_6-14Aryl, C_6-10Ara (C_1-6) Lucil or C_1-6Arca (C _6-10 ) Is ril; R^ThreeBut C_1-8Alkyl, C_3-8Cycloalkyl, C_2-8Alkenyl, C_2-8Alkynyl, C_6-14Aryl, C_6-10Ara (C_1-6) Lucil or C_1-6Arca (C _6-10 ) Is ril; R^FourBut C_6-10Aryl, or thienyl, benzo [b] thienyl, furyl, pyranyl, isobenzofuranyl, benzoxazolyl, 2H-pyrrolyl, pyrrolyl
, Imidazolyl, pyrazolyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazi
Nil, indolizinyl, isoindolyl, 3H-indolyl, indolyl, indol
Dazolyl, purinyl, 4H-quinolizinyl, isoquinolyl, quinolyl and tri
A heteroaryl group selected from the group consisting of azolyl;^FiveAnd R⁶But independently C_1-6Alkyl, C_1-10Ara (C_1-6) Lucil
Or C_1-6Arca (C_6-10) Is, or together with, the nitrogen atom to which they are attached, optionally substituted and optionally further substituted
Forming a 5- to 7-membered heterocyclic ring which may contain an oxygen atom or a nitrogen atom
The method according to claim 1, wherein Is 18. R ^1, C _1-6 alkyl, C _3-6 cycloalkyl, or C _{6 -10} aryl,; R ² is, Ri Oh methyl, ethyl, propyl, butyl, methoxy or ethoxy, R ³ is methyl, ethyl, tert-butyl, or benzyl; R ⁴ is phenyl or phenyl substituted with halogen, C _1-6 alkyl, C _1-6 alkoxy, carboxy or amino. NR ⁵ R ⁶ is dimethylamino, diethylamino, pyrrolidino, piperidino substituted with halogen, C _1-6 alkyl, C _6-10 aralkyl (C _1-6 ) alkyl, C _1-6 alkoxy, carboxy or amino; 18. The method according to claim 17, wherein the compound is one of, morpholino, and oxazolidinone. (A) Formula I: (B) treating the substituted aryl or heteroaryl oxazoline with a strong base to deprotonate the substituted aryl or heteroaryl oxazoline to form an enolate; and (b) converting the enolate to titanium, aluminum, tin, zinc , Magnesium and boron, and then transmetalated with a metal selected from the formula X
IV: Wherein R ¹ is alkyl, alkenyl, alkynyl, cycloalkyl, aryl, alkaryl, or aralkyl for each of Formula I, Formula II, and Formula XIV above (wherein aryl, aralkyl, or alkaryl R ² is alkyl, cycloalkyl, aryl, alkaryl, aralkyl, alkoxy, hydroxy, alkoxyalkyl, or amide (wherein the aryl, aralkyl or R ³ is alkyl, alkenyl, alkynyl, cycloalkyl, aryl, or alkaryl (any of which may be optionally substituted) R ⁴ is optionally substituted Aryl or optionally substituted heteroaryl; R ⁵ and R ⁶ are independently one of alkyl or alkaryl; or R ⁵ and R ⁶ are Together form a 5- to 7-membered heterocyclic ring which is optionally substituted and may optionally contain further oxygen or nitrogen atoms. Comprising reacting with the indicated formylamide, a compound of formula II: A method for forming a substituted oxazoline compound represented by the formula: 20. Furthermore, the oxazoline compound of the formula II is catalytically hydrogenated, after which the reaction mixture obtained if desired is refluxed, whereby the formula IV: 20. The method according to claim 19, comprising forming a β-lactam represented by the formula: wherein R ¹ , R ² and R ³ are the same as defined in claim 19. 21. The method further comprises saponifying a compound of formula IV,
Upon cyclization, a compound of formula VII: [In the formula, R ¹ and R ² are the same as defined in claim 19] crust having a - method according to claim 20, wherein the includes forming a lactacystin β- lactone compound. 22. (a) Formula XV: An asymmetric dihydroxylation of the alkene intermediate of formula XVIa: (B) reacting the optically active diol of formula XVIa with an orthoester under an acid catalyst to obtain a mixed orthoester; Acyl halide, HCl, HBr, HI, Me ₃ SiCl, Me ₃ Si
Reacting with a reagent selected from the group consisting of I, Me ₃ SiBr and a halogen-containing Lewis acid to form a compound of formula XVIIa: Wherein X is Cl, Br or I; (c) reacting the haloester derivative with an alkali metal azide to form a compound of formula XVI
IIa: embedded image (D) hydrogenating the azide to form a compound of formula XIXa: (E) subjecting the compound of formula XIXa to ring closure conditions to give a compound of formula Ia: Wherein R ¹ is alkyl, alkenyl, alkynyl, cycloalkyl, aryl, alkaryl, or aralkyl, wherein the aryl, aralkyl, or alkaryl ring moiety is optionally substituted; ³ is alkyl, cycloalkyl, aryl, or alkaryl (all of which are optionally substituted); R ⁴ is optionally substituted aryl or optionally substituted Wherein, for each of the above formulas XV, XVIa, XVIIa, VIIIa and XIXa, R ¹ , R ³ and R ⁴ are the same as in the definition of formula Ia. To form a substituted aryl oxazoline compound of formula Ia Law. 23. In the step (a), a dihydroxylation reaction is carried out using AD-mix-β in the presence of methanesulfonamide to obtain a diol represented by the formula XVIa in a configuration-selective manner. The method according to claim 22, characterized in that: 24. The process according to claim 22, wherein in step (a), the dihydroxylation reaction is carried out using N-oxide as a reoxidant. 25. In step (b), the diol of formula XVIa is treated with an orthoester in the presence of a Lewis or Bronsted acid catalyst to give a mixed orthoester, which is treated with acetyl bromide. With formula XVIIa (wherein
X is Br) in the in situ
23. The method according to claim 22, wherein the conversion is performed by: 26. The method according to claim 25, wherein the orthoester used in the reaction is an aromatic carboxylic acid orthoester. 27. The method according to claim 26, wherein said orthoester is trimethylorthobenzoic acid. 28. The method according to claim 25, wherein the acid catalyst is HBr, SnCl ₄ , TiCl ₄ , BBr ₃ or boron trifluoride. 29. In step (c), the crude haloester of formula XVIIa is converted to the azide of formula XVIIIa by treatment with an alkali metal azide in a polar aprotic organic solvent. Claim 2
2. The manufacturing method according to 2. 30. The process according to claim 22, wherein in step (d), the catalytic hydrogenation of the azide of formula XVIIIa is carried out over a palladium catalyst in ethyl acetate. 31. The process according to claim 30, wherein the catalytic hydrogenation proceeds with the simultaneous transfer of an aroyl group to obtain a hydroxyamide represented by the formula XIXa. 32. In step (e), the hydroxyamide of formula XIXa is treated with thionyl chloride in methylene chloride to effect ring closure with hydroxyl inversion to give the cis-substituted oxazoline of formula Ia 23. The method according to claim 22, wherein 33. The process of claim 32, wherein said cis-oxazoline is converted to trans-oxazoline under equilibrium conditions by reversing the configuration of the ester substituent. 34. (a) Formula XV: Asymmetric dihydroxylation of the alkene intermediate of formula XVIb: (B) reacting the optically active diol of formula XVIb with an orthoester under an acid catalyst to obtain a mixed orthoester; With a reagent selected from the group consisting of acyl halides, HCl, HBr, HI, Me ₃ SiCl, Me ₃ SiI, Me ₃ SiBr, and halogen-containing Lewis acids to form a compound of formula XVIIb: Wherein X is Cl, Br or I; (c) reacting the haloester derivative with an alkali metal azide to form a compound of formula XVI
IIb: embedded image (D) hydrogenating the azide to form a compound of formula XIXb: (E) subjecting the compound of formula XIXb to ring closure conditions to give the compound of formula Ib: Wherein R ¹ is alkyl, alkenyl, alkynyl, cycloalkyl, aryl, alkaryl, or aralkyl, wherein the aryl, aralkyl, or alkaryl ring moiety is optionally substituted; ³ is alkyl, cycloalkyl, aryl, or alkaryl (all of which are optionally substituted); R ⁴ is optionally substituted aryl or optionally substituted [Wherein, for each of the above formulas XV, XVIb, XVIIb, XVIIIb and XIXb, R ¹ , R ³ and R ⁴ are the same as defined in formula Ib] Forming a substituted aryl oxazoline compound of Formula Ib comprising forming the compound. A manufacturing method in which. 35. In the step (a), a dihydroxylation reaction is carried out using AD-mix-α in the presence of methanesulfonamide to obtain a diol represented by the formula XVIb in a configuration-selective manner. 35. The method according to claim 34, wherein: 36. The process according to claim 34, wherein in the step (a), the dihydroxylation reaction is carried out using N-oxide as a reoxidant. 37. In step (b), the diol of formula XVIb is treated with an orthoester under a Lewis or Bronsted acid catalyst to obtain a mixed orthoester, which is treated with acetyl bromide. Formula XVIIb, wherein X
35. The method according to claim 34, wherein the compound is converted into a haloester represented by the formula: 38. The method according to claim 37, wherein the orthoester used in the reaction is an aromatic carboxylic acid orthoester. 39. The method according to claim 38, wherein said orthoester is trimethylorthobenzoic acid. 40. The method according to claim 37, wherein the acid catalyst is HBr, SnCl ₄ , TiCl ₄ , BBr ₃ or boron trifluoride. 41. In step (c), the crude haloester of formula XVIIb is converted to the azide of formula XVIIIb by treatment with an alkali metal azide in a polar aprotic organic solvent. The method according to claim 34. 42. The process according to claim 34, wherein in step (d), the catalytic hydrogenation of the azide of formula XVIIIb is carried out over a palladium catalyst in ethyl acetate. 43. The catalytic hydrogenation proceeds with simultaneous transfer of an aroyl group,
43. The process according to claim 42, wherein a hydroxyamide of formula XIXb is obtained. 44. In step (e), the hydroxyamide of formula XIXb is treated with thionyl chloride in methylene chloride to effect ring closure with hydroxyl inversion to give the trans-substituted oxazoline of formula Ib 35. The method according to claim 34, wherein 45. A compound of formula XVIIIa: Comprising hydrogenating an azide compound having the formula XIXa: Wherein, for each of the above formulas XVIIIa and XIXa, R ¹ is alkyl, alkenyl, alkynyl, cycloalkyl, aryl, alkaryl, or aralkyl (wherein any ring moiety of the aryl, aralkyl, or alkaryl is desired) R ³ is alkyl, cycloalkyl, aryl, or alkaryl (wherein any of these may be optionally substituted); R ⁴ is aryl or heteroaryl (Wherein any of these may be optionally substituted)]. 46. The compound of formula XIXa is further subjected to ring closure conditions to give a compound of formula Ia: [Wherein, R ^1, R ³ and R ⁴ are the same as defined in claim 45] A method according to claim 45, wherein the includes forming a substituted oxazoline compound represented by the. 47. A compound of formula XVIIIb: Comprising hydrogenation of an azide compound having the formula XIXb: [Wherein for each of the above formulas XVIIIb and XIXb, R ¹ is alkyl, alkenyl, alkynyl, cycloalkyl, aryl, alkaryl, or aralkyl (wherein any ring moiety of the aryl, aralkyl, or alkaryl is desired) R ³ is alkyl, cycloalkyl, aryl, or alkaryl (wherein any of these may be optionally substituted); R ⁴ is aryl or heteroaryl (Wherein any of these may be optionally substituted)]. 48. The compound of formula XIXb is further subjected to ring closure conditions to give a compound of formula Ib: Wherein, R ^1, R ³ and R ⁴ are the same as defined in claim 47] includes forming a substituted oxazoline compound represented by 47. A process according. 49. Formula VI or VII: Wherein R ¹ is C _1-12 alkyl, C _3-8 cycloalkyl, C _2-8 alkenyl, C _2-8 alkynyl, C _6-14 aryl, C _6-10 ar (C _1-6 ) alkyl , or a C _1-6 alk (C _6-10) Lil; R ² is a C _2-6 alkyl; R ⁷ is an alkyl, aryl, aralkyl or alkaryl (herein, the alkyl, aryl , Aralkyl or alkaryl may be optionally substituted)], or a salt thereof. 50. The compound according to claim 49, wherein R ¹ is C _1-4 alkyl. 51. The compound according to claim 50, wherein R ¹ is isopropyl. 52. The compound of claim 49, wherein R ² is ethyl, n-propyl, n-butyl, or isobutyl. 53. The compound according to claim 52, wherein R ² is ethyl. 54. The compound according to claim 52, wherein R ² is n-propyl. 55. The compound according to claim 52, wherein R ² is n-butyl. 56. The compound according to claim 52, wherein R ² is isobutyl. 57. Formula XIV: Wherein R ² is C _1-8 alkyl, C _3-8 cycloalkyl, C _2-8 alkenyl, C _2-8 alkynyl, C _6-14 aryl, C _6-10 ara (C _1-6 ) alkyl or C _1-6 a alk (C _{6-1 0)} Lil,; R ⁵ and R ⁶ are independently, C _1-6 alkyl, C _6-10 Ara (C _1-6) alkyl, Or C _1-6 alk (C _6-10 ) lyl, or together with the nitrogen atom to which they are attached, may be optionally substituted and optionally further oxygen or nitrogen atoms To form a 5- to 7-membered heterocyclic ring, which may contain: or a salt thereof. 58. A pharmaceutical composition comprising a compound according to any one of claims 49 to 56 and a pharmaceutically acceptable carrier or diluent. 59. Formula II: embedded image Wherein R ¹ is alkyl, alkenyl, alkynyl, cycloalkyl, aryl, alkaryl, or aralkyl, wherein any ring portion of the aryl, aralkyl, or alkaryl is optionally substituted. R ² is alkyl, cycloalkyl, aryl, alkaryl, aralkyl, alkoxy, hydroxy, alkoxyalkyl, or amide, wherein any ring moiety of the aryl, aralkyl or alkaryl is optionally substituted; R ³ is alkyl, alkenyl, alkynyl, cycloalkyl, aryl, or alkaryl (all of which may be optionally substituted); R ⁴ is optionally substituted aryl or An optionally substituted heteroaryl Or when, R ⁵ and R ⁶ taken together with the nitrogen atom to which they are attached; a Lumpur; R ⁵ and R ⁶ are independently one at either of the alkyl or alkaryl To form a 5- to 7-membered heterocyclic ring which may be optionally substituted and optionally contain an additional oxygen or nitrogen atom, or a salt thereof. 60. wherein R ¹ is, C _1-12 alkyl, C _3-8 cycloalkyl, C _2-8 A alkenyl, C _2-8 alkynyl, C _6-14 aryl, C _6-10 Ara (C _{1- 6)} alkyl, was or is C _1-6 alk (C _6-10) Lil (where the aryl, aralkyl or may be optionally substituted any of the ring portion of the alkaryl); R ² Is a C _1-8 alkyl, C _3-8 cycloalkyl, C _2-8 alkenyl, C _2-8 alkynyl, C _6-14 aryl, C _6-10 ar (C _1-6 ) alkyl, or C _{1 -6} alk (C _{6 -10)} a drill (wherein the aryl, any ring moiety of the aralkyl or alkaryl may also be optionally substituted); R ³ is, C _1-8 alkyl, C _{3- 8} cycloalkyl, C _2-8 alkenyl, C _2-8 Al Kiniru, C _6-14 aryl, C _6-10 Ara (C _1-6) alkyl or C _1-6 alk (C _{6- 10),} A le (here, may be optionally substituted none of these); R ⁴ is optionally optionally substituted C _6-10 aryl or thienyl, benzo [beta] thienyl, furyl, Pyranyl, isobenzofuranyl, benzoxazolyl, 2H-pyrrolyl, pyrrolyl, imidazolyl, pyrazolyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, indolizinyl, isoindolyl, 3H
- indolyl, indolyl, indazolyl, purinyl, 4H-quinolizinyl, isoquinolyl, an optionally optionally substituted heteroaryl group selected from the group consisting of quinolyl and triazolyl; R ⁵ and R ^6, independently , C _1-6 alkyl, C _6-10 Ara (C _1-6) alkyl, or was or is a C _1-6 alk (C _6-10) Lil, or together with the nitrogen atom to which it is attached 60. The compound of claim 59, which forms a 5- to 7-membered heterocycle optionally substituted and optionally containing an additional oxygen or nitrogen atom. . 61. Formula III: Wherein R ¹ is alkyl, alkenyl, alkynyl, cycloalkyl, aryl, alkaryl, or aralkyl, wherein any ring portion of the aryl, aralkyl, or alkaryl is optionally substituted. R ² is alkyl, cycloalkyl, aryl, alkaryl, aralkyl, alkoxy, hydroxy, alkoxyalkyl, or amide, wherein any ring moiety of the aryl, aralkyl or alkaryl is optionally substituted; R ³ is alkyl, alkenyl, alkynyl, cycloalkyl, aryl, or alkaryl (wherein any of these may be optionally substituted); R ⁵ and R ⁶ are independently Is one of alkyl or alkaryl; or When R ⁵ and R ⁶ taken together with the nitrogen atom to which they are attached, optionally may be substituted, and may 5 not also contain additional oxygen atom or nitrogen atom optionally 7- membered Or a salt thereof. 62. wherein R ¹ is, C _1-12 alkyl, C _3-8 cycloalkyl, C _2-8 A alkenyl, C _2-8 alkynyl, C _6-14 aryl, C _6-10 Ara (C _{1- 6)} alkyl, was or is C _1-6 alk (C _6-10) Lil (where the aryl may be optionally substituted any of the ring moiety of the aralkyl or alkaryl); R ² is , C _1-8 alkyl, C _3-8 cycloalkyl, C _2-8 alkenyl, C _2-8 alkynyl, C _6-14 aryl, C _6-10 ar (C _1-6 ) alkyl, or C _1- alkyl. ₆ alk (C _{6- 10)} is Lil (where the aryl, any ring moiety of the aralkyl or alkaryl may also be optionally substituted); R ³ is, C _1-8 alkyl, C _{3- 8} cycloalkyl, C _2-8 alkenyl, C _2-8 Al Kiniru, C _6-14 aryl, C _6-10 Ara (C _1-6) alkyl or C _1-6 alk (C _{6- 10)} Li, A in the (here, may be any of those optionally substituted); R ⁵ and R ^6, independently, C _1-6 alkyl, C _6-10 Ara (C _1-6) alkyl Or a C _1-6 alk (C _6-10 ) lyl, or together with the nitrogen atom to which they are attached, may be optionally substituted and optionally further oxygen atoms or 63. The compound according to claim 61, which forms a 5- to 7-membered heterocycle optionally containing a nitrogen atom. 63. A compound of formula XVIIa or XVIIb: Wherein R ¹ is alkyl, alkenyl, alkynyl, cycloalkyl, aryl, alkaryl, or aralkyl, wherein any ring portion of the aryl, aralkyl, or alkaryl is optionally substituted. ; R ² is Cl, Br, or I; R ³ is alkyl, alkenyl, alkynyl, cycloalkyl, aryl, a or alkaryl (may be optionally substituted none of these); R ⁴ is Or an optionally substituted aryl or an optionally substituted heteroaryl], or a salt thereof. 64. A compound of formula XVIIIa or XVIIIb: Wherein R ¹ is alkyl, alkenyl, alkynyl, cycloalkyl, aryl, alkaryl, or aralkyl (wherein any ring portion of the aryl, aralkyl or alkaryl may be optionally substituted R ³ is alkyl, alkenyl, alkynyl, cycloalkyl, aryl, or alkaryl (wherein any of these may be optionally substituted); R ⁴ may be optionally substituted A good aryl or an optionally substituted heteroaryl], or a salt thereof. 65. (a) Formula VIII: Wherein R ⁸ is isopropyl or benzyl. The anion of the compound of formula IX is allylated with R ² CH ₂ COCl to give a compound of formula IX: Wherein R ² and R ⁸ are the same as defined below and above, respectively, to form an acyloxazolidinone represented by the formula: (b) a stereoconfiguration of the acyloxazolidinone represented by the formula IX and benzyloxymethyl chloride Selectively reacting to form a compound of formula X: Wherein R ² and R ⁸ are each as defined below and as defined above; (c) hydrolyzing the protected alcohol of formula X to form a protected alcohol of formula XI: Embedded image Wherein R ² is the same as defined below; and (d) coupling the acid of formula XI with the amine R ⁵ R ⁶ NH ₂ to form a compound of formula XII : Wherein R ² , R ⁵ and R ⁶ are each the same as defined below; to obtain an amide represented by the formula: (e) catalytic hydrogenation of the amide represented by the formula XII to obtain a compound represented by the formula XIII: Scheme 44 Wherein R ² , R ⁵ and R ⁶ are each as defined below; and (f) oxidizing the resulting alcohol of formula XIII to form an alcohol of the formula XIV: [R ² is alkyl, cycloalkyl, aryl, alkaryl, aralkyl, alkoxy, hydroxy, alkoxyalkyl, or amide (wherein any ring portion of the aryl, aralkyl, or alkaryl may be optionally substituted. R ⁵ and R ⁶ are independently one of alkyl or alkaryl; or R ⁵ and R ⁶ , when taken together with the nitrogen atom to which they are attached,
To form a 5- to 7-membered heterocyclic ring which is optionally substituted and may optionally contain additional oxygen or nitrogen atoms. A process for forming an enantiomer-rich formylamide of the formula XIV, or a salt thereof. 66. A method wherein R ² is C _1-8 alkyl, C _3-8 cycloalkyl, C _2-8 alkenyl, C _2-8 alkynyl, C _6-14 aryl, C _6-10 ara (C _1-6 ) alkyl, or a C _1-6 alk (C _6-10) Lil; R ⁵ and R ^6, independently, C _1-6 alkyl, C _6-10 Ara (C _1-6) alkyl Or a C _1-6 alk (C _6-10 ) lyl, or together with the nitrogen atom to which they are attached, may be optionally substituted and optionally further oxygen or 66. The process according to claim 65, wherein a 5- to 7-membered heterocycle optionally containing a nitrogen atom is formed. 67. A method for inhibiting proteasome function in a cell, comprising contacting the cell with the compound according to claim 49. 68. A method for inhibiting proteasome function in a mammal, comprising administering the compound according to claim 49 to the mammal in an amount effective to inhibit proteasome function. 69. A method of treating inflammation comprising administering to a subject an anti-inflammatory effective amount of the compound of claim 49. 70. A method of treating cancer comprising administering to a subject an anti-tumor or anti-metastatic effective amount of a compound according to claim 49. 71. A method of treating ischemia or reperfusion injury in a mammal, comprising administering to the mammal an effective amount of the compound of claim 49. 72. The method of claim 71, wherein said ischemia is the result of a vascular occlusion. 73. The method of claim 71, wherein said vascular obstruction occurs during a stroke.