AU2002351170B8 - Production of chirally pure alpha-amino acids and N-sulfonyl alpha-amino acids - Google Patents
Production of chirally pure alpha-amino acids and N-sulfonyl alpha-amino acids Download PDFInfo
- Publication number
- AU2002351170B8 AU2002351170B8 AU2002351170A AU2002351170A AU2002351170B8 AU 2002351170 B8 AU2002351170 B8 AU 2002351170B8 AU 2002351170 A AU2002351170 A AU 2002351170A AU 2002351170 A AU2002351170 A AU 2002351170A AU 2002351170 B8 AU2002351170 B8 AU 2002351170B8
- Authority
- AU
- Australia
- Prior art keywords
- substituted
- sulfonyl
- amino acid
- alkyl
- phenyl
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Ceased
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D307/00—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom
- C07D307/02—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom not condensed with other rings
- C07D307/34—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom not condensed with other rings having two or three double bonds between ring members or between ring members and non-ring members
- C07D307/56—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom not condensed with other rings having two or three double bonds between ring members or between ring members and non-ring members with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
- C07D307/64—Sulfur atoms
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
- A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D333/00—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom
- C07D333/02—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom not condensed with other rings
- C07D333/04—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom not condensed with other rings not substituted on the ring sulphur atom
- C07D333/26—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom not condensed with other rings not substituted on the ring sulphur atom with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
- C07D333/30—Hetero atoms other than halogen
- C07D333/34—Sulfur atoms
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D409/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms
- C07D409/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms containing two hetero rings
- C07D409/12—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms containing two hetero rings linked by a chain containing hetero atoms as chain links
Landscapes
- Organic Chemistry (AREA)
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Neurosurgery (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biomedical Technology (AREA)
- Neurology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Life Sciences & Earth Sciences (AREA)
- Hospice & Palliative Care (AREA)
- General Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Pharmacology & Pharmacy (AREA)
- Psychiatry (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Description
WO 03/050062 PCT/US02/38117 PRODUCTION OF CHIRALLY PURE a-AMINO ACIDS AND N-SULFONYL a-AMINO ACIDS BACKGROUND OF THE INVENTION 5 This invention relates to a novel process of producing chirally pure a-amino acids and N-sulfonyl a-amino acids. Compounds of the present invention are useful for a variety of purposes, including for use in pharmaceutical compositions. A variety of techniques have been described for production of a preferred enantiomer from a-amino acids. More efficient means for producing chirally pure 10 target compounds are needed. SUMMARY OF THE INVENTION In one aspect, the present invention comprises a process for preparing chirally pure S-enantiomers of a-amino acids. 15 In a further aspect, a process is provided for preparing chirally pure S enantiomers of 2-aminoalcohols, aldehydes and oximes. In yet another aspect, a process is provided for preparing chirally pure S enantiomers of N-sulfonyl a-amino acids. In a further aspect, a process is provided for preparing chirally pure N-sulfonyl 20 2-aminoalcohols, aldehydes and oximes. These and other aspects of the invention will be apparent to one of skill in the art upon reading of the following detailed description of the invention. DETAILED DESCRIPTION OF THE INVENTION 25 In one aspect, the present invention is directed to a process for the preparation of chiral a-amino acids. In another aspect, the present invention provides a process for the resolution of chiral N-sulfonyl a-amino acids. Both processes of the invention produce chirally pure compounds which can 30 be converted to suitable target compounds, including the corresponding 2 1 WO 03/050062 PCT/US02/38117 aminoalcohols or N-sulfonyl 2-aminoalcohols, aldehydes and oximes, among other desirable target compounds. As used herein, the term "chirally pure" refers to compounds which are in greater than about 95% S-enantiomeric form, preferably greater than about 97%, as 5 measured by chiral high performance liquid chromatography (HPLC). Other methods of measuring chiral purity include conventional analytical methods, including specific rotation, and conventional chemical methods. However, the technique used to measure chiral purity is not a limitation on the present invention. As used herein, the term "pharmaceutically useful" refers to compounds 10 having a desired biological effect, whether as a therapeutic, immune stimulant or suppressant, adjuvant, or vaccinal agent. Similarly, a variety of compounds which are suitable for use in non-pharmaceutical applications, e.g., a diagnostic, a marker, among others may be produced by the method of the invention. However, other pharmaceutically useful compounds may be produced by this method. 15 The compounds produced by the present invention and any target compounds into which they are converted can be used in the form of salts derived from pharmaceutically or physiologically acceptable acids or bases. These salts include, but are not limited to, the following salts with organic and inorganic acids such as acetic, lactic, citric, tartaric, succinic, fumaric, maleic, malonic, mandelic, mallic, 20 hydrochloric, hydrobromic, phosphoric, nitric, sulfuric, methanesulfonic, toluenesulfonic and similarly known acceptable acids, and mixtures thereof. Other salts include salts with alkali metals or alkaline earth metals, such as sodium (e.g., sodium hydroxide), potassium (e.g., potassium hydroxide), calcium or magnesium. These salts, as well as other compounds produced by the method of the 25 invention may be in the form of esters, carbamates and other conventional "pro-drug" forms, which, when administered in such form, convert to the active moiety in vivo. In one desirable embodiment, the prodrugs are esters. See, e.g., B. Testa and J. Caldwell, "Prodrugs Revisited: The "Ad Hoc" Approach as a Complement to Ligand Design", Medicinal Research Reviews, 16(3):233-241, ed., John Wiley & Sons 30 (1996). Both natural and unnatural o-amino acids, natural and unnatural 2 aminoalcohols, and intermediates thereof, may be prepared according to the present 2 WO 03/050062 PCT/US02/38117 invention. Typically, a-amino acids are characterized by the formula
(NH
2 )(CHR.)(COOH), in which R. is an aliphatic radical. The a-amino acids prepared according to the invention can be converted to N-sulfonyl a-amino acids and other desired compounds. Such other desired compounds include, without limitation, 5 the corresponding 2-aminoalcohols, aldehydes, oximes, and pharmaceutically acceptable salts, hydrates, and prodrugs thereof Similarly, both natural and unnatural N-sulfonyl a-amino acids, natural and unnatural N-sulfonyl 2-aminoalcohols, and intermediates thereof, may be prepared according to the present invention. Thus, the N-sulfonyl a-amino acids described herein can be readily reduced to 2 10 aminoalcohols, or converted to the corresponding aldehydes, oximes, and pharmaceutically acceptable salts, hydrates, and prodrugs thereof, using techniques known to those of skill in the art. For example, chirally pure a-amino acids produced according to the method of the invention and having the formula (R) 2
CH(CH
2 )nCH(CO 2 H)NH-R' can readily be 15 converted to chirally pure 2-aminoalcohols. In another example, chirally pure N sulfonyl a-amino acids produced according to the invention and having the formula
(R)
2
CH(CH
2 )nCH(CO 2
H)NH-S(O)
2 R' are readily converted to N-sulfonyl 2 aminoalcohols of the formula (R) 2
CH(CH
2 )nCH(CH 2
OH)NHS(O)
2 R'. Suitably, in the above formulae, n is 0 to about 10; R is lower alkyl, substituted lower alkyl, lower 20 alkenyl, substituted lower alkenyl, lower alkynyl, substituted lower alkynyl, cycloalkyl, substituted cycloalkyl, phenyl, substituted phenyl, benzyl, substituted benzyl, CH 2 cycloalkyl, CH 2 -3-indole, CH(loweralkyl)-2-furan, CH(loweralkyl)-4 methoxyphenyl, CH(loweralkyl)phenyl, or CH(OH)-4-SCH 3 -phenyl; and R' is selected from among H, lower alkyl, substituted lower alkyl, lower alkenyl, 25 substituted lower alkenyl, lower alkynyl, substituted lower alkynyl, heterocycle, substituted heterocycle, phenyl, substituted phenyl, benzyl, substituted benzyl, cycloalkyl, and substituted cycloalkyl, among other suitable groups. In another example, an N-sulfonyl 2-aminoalcohol having the formula
(R)
2
CH(CH
2 )nCH(CH 2 OH)NH-S(0) 2 -2-C 4
H
2 S-5-Cl is prepared using the method of 30 the invention. However, the chirally pure compounds produced by the methods of the present invention are not limited by the above formulae. 3 WO 03/050062 PCT/US02/38117 The term "alkyl" is used herein to refer to both straight- and branched-chain saturated aliphatic hydrocarbon groups having one to ten carbon atoms, preferably one to eight carbon atoms and, most preferably, one to six carbon atoms; "alkenyl" is intended to include both straight- and branched-chain alkyl groups with at least one 5 carbon-carbon double bond and two to eight carbon atoms, preferably two to six carbon atoms; "alkynyl" group is intended to cover both straight- and branched-chain alkyl groups with at least one carbon-carbon triple bond and two to eight carbon atoms, preferably two to six carbon atoms. As used herein, the term "lower" refers to any of the above-defined groups having one to six carbon atoms. 10 The terms "substituted alkyl", "substituted alkenyl", "substituted alkynyl", "substituted lower alkyl", "substituted lower alkenyl", "substituted lower alkynyl" refer to alkyl, alkenyl, alkynyl, lower alkyl, lower alkenyl, and lower alkynyl as just described having from one to three substituents selected from the group including halogen, CN, OH, NO 2 , amino, aryl, heterocyclic, substituted aryl, substituted 15 heterocyclic, alkoxy, substituted alkoxy, aryloxy, substituted alkyloxy, alkylcarbonyl, alkylcarboxy, alkylamino, and arylthio. These substituents may be attached to any carbon of an alkyl, alkenyl, or alkynyl group provided that the attachment constitutes a stable chemical moiety. The term "aryl" is used herein to refer to a carbocyclic aromatic system, which 20 may be a single ring, or multiple aromatic rings fused or linked together as such that at least one part of the fused or linked rings forms the conjugated aromatic system. The aryl groups include, but are not limited to, phenyl, naphthyl, biphenyl, anthryl, tetrahydronaphthyl, phenanthryl, and indane. The term "substituted aryl" refers to aryl as just defined having one to four 25 substituents from the group including halogen, CN, OH, NO 2 , amino, alkyl, cycloalkyl, alkenyl, alkynyl, alkoxy, aryloxy, substituted alkyloxy, alkylcarbonyl, alkylcarboxy, alkylamino, and arylthio. The term "substituted benzyl" refers to a benzyl (Bn) group, having substituted on the benzene ring, one to five substituents from the group including 30 halogen, CN, OH, NO 2 , amino, alkyl, cycloalkyl, alkenyl, alkynyl, alkoxy, aryloxy, substituted alkyloxy, alkylcarbonyl, alkylcarboxy, alkylamino, and arylthio. 4 WO 03/050062 PCT/US02/38117 The term "heterocyclic" is used herein to describe a stable 4- to 7-membered monocyclic or a stable multicyclic heterocyclic ring which is saturated, partially unsaturated, or unsaturated, and which consists of carbon atoms and from one to four heteroatoms selected from the group including N, 0, and S atoms. The N and S 5 atoms may be oxidized. The heterocyclic ring also includes any multicyclic ring in which any of above defined heterocyclic rings is fused to an aryl ring. The heterocyclic ring may be attached at any heteroatom or carbon atom provided the resultant structure is chemically stable. Such heterocyclic groups include, for example, tetrahydrofuran, piperidinyl, piperazinyl, 2-oxopiperidinyl, azepinyl, 10 pyrrolidinyl, imidazolyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, oxazolyl, isoxazolyl, morpholinyl, indolyl, quinolinyl, thienyl, furyl, benzofuranyl, benzothienyl, thiamorpholinyl, thiamorpholinyl sulfoxide, isoquinolinyl, and tetrahydrothiopyran. The term "substituted heterocyclic" is used herein to describe the heterocyclic 15 groups just defined having one to four substituents selected from the group which includes halogen, CN, OH, NO 2 , amino, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, alkoxy, substituted alkoxy, aryloxy, substituted aryloxy, alkyloxy, substituted alkyloxy, alkylcarbonyl, substituted alkylcarbonyl, alkylcarboxy, substituted 20 alkylcarboxy, alkylamino, substituted alkylamino, arylthio, or substituted arylthio. The term "substituted cycloalkyl" is used herein to describe a carbon-based ring having more than 3 carbon-atoms which forms a stable ring and having from one to five substituents selected from the group consisting of halogen, CN, OH, NO 2 , amino, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, alkoxy, aryloxy, 25 substituted alkyloxy, alkylcarbonyl, alkylcarboxy, alkylamino, substituted alkylamino, arylthio, heterocyclic, substituted heterocyclic, aminoalkyl, and substituted aminoalkyl. Where the terms "substituted alkylcycloalkyl", "substituted alkylOBn", "substituted alkylpyridyl", "substituted alkylfuranyl", "substituted alkyl NHR 7 ", and 30 phenyl(substituted)alkyl, "substituted alkylOH", and "substituted alkylSRs" are recited in Formula I and Formula Ia below, the substitution may occur at the alkyl group or on the corresponding base compound. 5 WO 03/050062 PCT/US02/38117 As used in the definition of the R4 group in Formula I and Ia below, an N substituted piperidinyl group may be defined as a substituted heterocyclic group. Among particularly desirable substituents are N-alkyl-, N-aryl-, N-acyl-, and N sulfonyl piperidinyl groups. One particularly suitable N-acyl-piperidinyl group is N 5 t-butyloxycarbonyl (BOC)-piperidine. However, other suitable substituents can be readily identified by one of skill in the art. The term "alkoxy" is used herein to refer to the O(alkyl) group, where the point of attachment is through the oxygen-atom and the alkyl can be optionally substituted. The term "aryloxy" is used herein to refer to the O(aryl) group, where the 10 point of attachment is through the oxygen-atom and the aryl can be optionally substituted. The term "alkylcarbonyl" is used herein to refer to the CO(alkyl) group, where the alkyl can be optionally substituted and the point of attachment is through the carbon atom of the carbonyl group. The term "alkylcarboxy" is used herein to refer to the COO(alkyl) group, where the alkyl can be optionally substituted and the 15 point of attachment is through the carbon atom of the carboxy group. The term "aminoalkyl" refers to both secondary and tertiary amines wherein the alkyl or substituted alkyl groups, containing one to eight carbon atoms, which may be either same or different, and the point of attachment is on the nitrogen atom The term "halogen" refers to Cl, Br, F, or I. 20 The term "ring" structure, e.g., when R 3 and R 4 may form a ring structure in Formula I, includes a monocyclic structure, a bridged cyclo structure, and fused cyclo structures, unless the type of ring structure is otherwise specified. The term "strong non-nucleophilic base" refers to a non-nucleophilic basic reagent, which does not act as a nucleophile or bind to the reagents utilized according 25 to the reaction. A number of non-nucleophilic bases are known in the art and include sodium hydride, potassium hydride, lithium diisopropylamide and potassium hexamethyldisilazide. The term "aqueous base" refers to a solution composed of, at a minimum, a base and water. A number of bases which readily dissolve in water are known in the 30 art and include alkali metal hydroxides such as lithium hydroxide, sodium hydroxide or potassium hydroxide, among others. The aqueous base solution may further contain other reagents which do not interfere with the reactions of the present 6 invention, and include organic solvents such as tetrahydrofuran, methanol, ethanol, or hydrocarbon solvents, salts such as sodium chloride, and buffers, among others. The term "aqueous acid" refers to a solution composed of, at a minimum, an acid and water. The aqueous acid solution may further contain other reagents which do not 5 interfere with the reactions of the present invention. The term "strong acid" or "strong base" refers to an acid or base that is completely ionized in solution. Common strong acids include HCl, HBr, HI, HNO 3 , H 2
SO
4 , and HC10 4 . Common strong bases include hydroxides of the alkali metals (Li, Na, K, Rb, Cs) and hydroxides of the heavy alkaline earths (Ca, Sr, Ba). 10 The tent "inorganic" acid or "inorganic" base includes acids and bases which do not contain carbon. The term "organic solvent" may include any carbon-containing solvent known in the art, which does not react with the reagents utilized in the reaction and includes saturated hydrocarbon solvents, unsaturated hydrocarbon solvents, including aromatic 15 hydrocarbon solvents, alcohols, halocarbons, ethers, and acetates, among others. Throughout the description and claims of this specification, use of the word "comprise" and variations of the word, such as "comprising" and "comprises", is not intended to exclude other additives, components, integers or steps. The discussion of documents, acts, materials, devices, articles and the like is 20 included in this specification solely for the purpose of providing a context for the present invention. It is not suggested or represented that any or all of these matters formed part of the prior art base or were common general knowledge in the field relevant to the present invention as it existed in Australia before the priority date of each claim of this application. 25 Synthesis of Chirally Pure Compounds The chirally pure compounds can be prepared using the methods described below. Where reference to conventional techniques is made, one of skill in the art will be able to readily select appropriate synthetic methods and reagents, which are known in the 30 synthetic organic arts or variations of these methods by one skill ed in the art. See, generally, Comprehensive Organic Synthesis, "Selectivity, Strategy & Efficiency in Modem Organic Chemistry", ed., I. Fleming, Pergamon Press, New York (1991); Comprehensive Organic Chemistry, "The Synthesis and Reactions of Organic Compounds", ed. J.F. Stoddard, Pergamon Press, New York (1979). 7 Preparation of Chirally Pure a-Amino Acids In one aspect, the invention provides a method for preparing chirally pure a amino acids from chirally impure a-amino acids. For the preparation of chirally pure a amino acids XXXXVI, a novel asymmetric variant of the Strecker a-amino acid synthesis 5 is utilized (Scheme 14; J. Org. Chem. 54:1055-1062 (1989)). In this route (Scheme I), an aldehyde XXXXVII is reacted with a cyanide salt and a 10 15 20 25 30 7q WO 03/050062 PCT/US02/38117 methylbenzylamine or a salt thereof in a 1:1:1 molar ratio in a suitable solvent to afford the compound XXXXVIII. Included among desirable cyanide salts are sodium cyanide and potassium cyanide. However, other suitable cyanide salts may be readily selected for use in the method of the invention. Preferably, the solvent is 1:1 5 methanol to water. Suitably, the reaction is performed for about 12 to about 24 hours, and most preferably, about 18 hours. However, longer or shorter reaction times may be readily utilized. Optionally, following this reaction, a suspension containing, precipitate is formed, which is subjected to filtration and is washed (e.g., with water) to provide a powder. Compound XXXXVIII is dissolved with a strong inorganic acid 10 which is desirably cold upon combination with the compound (e.g., about 0'C to about 10'C) to provide the compound XXXXIX. Desirably, the strong inorganic acid is sulfuric acid. However, other strong inorganic acids may be readily selected. The reaction mixture is neutralized with an inorganic base and extracted with an organic solvent to compound XXXXIX. Suitably, extraction may utilize ethyl acetate or 15 another suitable compound, and further involves drying and concentrating to provide compound XXXXIX. The hydrogenolysis reaction takes place in the presence of a suitable catalyst under pressure, e.g., Pd or RaNi under 3 atm pressure, filtering to remove the catalyst followed by concentration to remove solvent provides compound XXXXX. Compound XXXXX is then dissolved with an aqueous acid to afford the 20 derivatives of formula XXXXVI. Where XXXXX has been dried to powder form, it is dissolved in a strong inorganic acid at high temperature to afford a salt of a chirally pure a-amino acid. For example, hydrochloric acid at 100 0 C may be utilized. Alternatively, other acids and other suitable temperatures may be readily selected by one of skill in the art. Most desirably, the hydrolysis step is performed over a period 25 of about 12 to 18 hours, or longer. In one suitable embodiment, the step is performed over 16 hours. Optionally, the resulting reaction mixture is concentrated to provide a product which consists of the amino acid salt and one equivalent of ammonium salt. In this example, the product is the amino acid hydrochloride salt and one equivalent of ammonium chloride. This product is dissolved in water to which the base, e.g., 30 sodium hydroxide or ammonium hydroxide, is added to form a solution. 8 Scheme 1 Q s 1. CN salt CN R H R HN )XXVOXOXVM H2N 0 H2N RHN XXXXIX 3. H 2 /catalyst 4. strong inorganic OH aid H 2 N o R H 2 N R H 2 N These chirally pure a-amino acids produced according to the method of the invention can be readily utilized in the form produced, or converted to a desired target compound. For example, a chirally pure a-amino acid can be readily converted to a chirally pure 2-aminoalcohol by reducing the a-amino acid to the 2-aminoalcohol and recrystallizing the 2-aminoalcohol to afford the chirally pure 2-aminoalcohol. These and other uses for the chirally pure a-amino acids of the invention will be readily apparent to those of skill in the art from the information provided herein and that known to those of skill in the art. In another aspect, the invention provides a process for resolving a chirally pure N-[(5-chloro-2-thienyl)sulfonyl]-3-ethylnorvaline for use in preparing a chirally pure a-amino acid, comprising the steps of: (a) forming a mixture of (+)-ephedrine hemihydrate and N-[(5 chloro-2-thienyl)sulfonyl]-3-ethylnorvaline in ethanol at a molar ratio of 1:1; (b) heating to at least 80'C to dissolve the solids; (c) cooling the mixture to allow it to form a precipitate; (d) washing the precipitate with ethanol and ethyl acetate to give diastereomeric salt; (e) recrystallizing the diastereomeric salt; (f) extracting the recrystallized diastereomeric salt to provide an organic extract; 9 (g) washing the organic extract, and (h) drying and optionally concentrating the organic extract to provide chirally pure N-[(5-chloro-2-thienyl)sulfonyl]-3-ethylnorvaline. In another aspect, the invention provides a method for producing chirally pure a-amino acids comprising the steps of: (a) reacting an aldehyde and potassium cyanide with an a methylbenzylamine or hydrochloride salt thereof and filtering to provide product (a); (b) reacting sulphuric acid and product (a); (c) neutralizing the acid from the reaction of step (b); (d) extracting product (c) from the neutralized acid; (e) hydrogenating product (d) in the presence of a catalyst; (f) mixing product (e) in hydrochloric acid to provide a salt of chirally pure a-amino acid; and (g) neutralizing the salt of chirally pure a-amino acid to provide the chirally pure a-amino acid. Preparation of Chirally Pure N-sulfonyl a-Amino Acids In another aspect, the invention provides a scheme for resolving a chirally impure N-sulfonyl a-amino acid having a p-branched alkyl substituent to provide a chirally pure N-sulfonyl a-amino acid. Desirably, the N-sulfonyl a-amino acid is N sulfonyl p-ethylnorvaline. In another embodiment, the norvaline compound can be substituted with a compound selected from among N-sulfonyl O-ethylnorvaline, N sulfonylvaline, and N-sulfonyl 0-n-propylnorleucine. Alternatively, one ofskill in the art may use the method of the invention with another selected N-sulfonyl a-amino 9a WO 03/050062 PCT/US02/38117 acid having a p-branched alkyl substituent for preparing the corresponding chirally pure compound. Suitably, N-sulfonyl p-ethylnorvaline (or another selected compound) is mixed with chirally pure ephedrine heniihydrate in ethanol at a molar ratio of 1:1. 5 The mixture is then heated to dissolve the solids. In one embodiment, the mixture is heated to about 80'C. However, other suitable temperatures may be readily selected. Thereafter, the mixture is cooled in order to allow a precipitate to form This cooling step may be performed at room temperature or at reduced temperature (e.g., about 5 C) overnight (about 16 - 20 hours). The temperature and the period of the cooling 10 step each may be adjusted upwardly or downwardly, as needed or desired. Optionally, the suspension is filtered or washed following cooling. The salt is then recrystallized and then dissolved in a solvent and a strong aqueous acid. Suitably, the recrystallizing step is performed in boiling ethyl acetate and the recrystallized salt is separated. This may be performed using filtration or other conventional methods. 15 Suitably, the salt is dissolved in an organic solvent and strong aqueous acid. The organic extract is washed, dried and concentrated to provide the chirally pure N sulfonyl a-amino acid. In one embodiment, the wash step is performed with a strong aqueous acid such as, for example, hydrochloric acid, and drying is performed with sodium sulfate or the like. 20 Suitably, these chirally pure a-amino acids and N-sulfonyl a-amino acids are useful for a variety of purposes. For example, these chirally pure a-amino acids and N-sulfonyl a-amino acids can be converted to the corresponding N-sulfonyl 2 aminoalcohols by the methods described herein. Thus, in one embodiment the chirally pure a-amino acids produced according 25 to the present invention are useful in the synthesis of chiral N-sulfonyl a-amino acids. Suitable methods for preparation of these chiral N-sulfonyl a-amino acids are provided herein. 10 WO 03/050062 PCT/US02/38117 Conversion to 2-Aminoalcohols The processes of the invention provide efficient routes to the synthesis of chirally pure S enantiomers of a-amino acids and N-sulfonyl a-amino acids which are useful in preparing 2-aminoalcohols or N-sulfonyl 2-aminoalcohols, and 5 intermediates thereof, which are useful for a variety of purposes. For example, the exemplary compounds provided herein, the N-sulfonyl 2 amino alcohols and their corresponding aldehydes, oximes and salts, are useful for modulating p-anyloid production, which is implicated in amyloid angiopathy, cerebral amyloid angiopathy, systemic amyloidosis, Alzheimer's Disease (AD), 10 hereditary cerebral hemorrhage with amyloidosis of the Dutch type, inclusion body myositis, Down's syndrome, among others. Thus, the compounds of Formula (I) are useful in modulating beta amyloid production in subjects at risk for, or suffering from, AD or other diseases resulting from elevated levels of beta amyloid protein in the brain. These compounds and their uses are described in more detail in co-pending US 15 Patent Application No. 10/014,304, filed December 11, 2001, which is incorporated herein by reference. The compounds of Formula (I) include pharmaceutically acceptable salts and/or hydrates or prodrugs thereof, wherein Formula (I) is: 0 R 5 Re NT R6- 1-IWfIIlX X O
R
3 (I) 20 R 3 is selected from the group consisting of hydrogen, alkyl, and substituted alkyl;
R
4 is selected from the group consisting of hydrogen, alkyl, substituted alkyl, alkylcycloalkyl, substituted alkylcycloalkyl, phenyl(substituted)alkyl, alkylOH, substituted alkylOH, alkylOBn, substituted alkylOBn, alkylpyridyl, substituted 25 alkylpyridyl, alkylfuranyl, substituted alkylfuranyl, CH(OH)phenyl, CH(OH)substituted phenyl, alkenyl, substituted alkenyl, cycloalkyl, substituted cycloalkyl, N-substituted-piperidinyl, piperidinyl, substituted piperidinyl, 11 WO 03/050062 PCT/US02/38117 tetrahydrothiopyran, substituted tetrahydrothiopyran, 2-indane, substituted 2-indane, phenyl, substituted phenyl, alkylNHR 7 , and substituted alkylNHR 7 ; with the proviso that R 3 and R are not both hydrogen;
R
7 is alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, benzyl, 5 substituted benzyl, alkylOH, substituted alkylOH, alkylSRs, or substituted alkzylSRs;
R
8 is alkyl, substituted alkyl, benzyl, or substituted benzyl; or R 3 and R4 may be joined to form a ring;
R
5 is selected from the group consisting of hydrogen, lower alkyl, substituted lower alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, CH 2 cycloalkyl, 10 substituted CH 2 cycloalkyl, benzyl, substituted benzyl, and CH 2
CH
2 QR9; Q is 0, NH or S;
R
9 is lower alkyl, substituted lower alkyl, phenyl, or substituted phenyl;
R
6 is selected from the group consisting of hydrogen, halogen and CF 3 ; T is selected from the group consisting of OH 0 RH, and - HC -N OH 15
R
1 and R 2 are independently selected from the group consisting of hydrogen, alkyl, substituted alkyl, CF 3 , alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl, substituted cycloalkyl, phenyl, substituted phenyl, and (CH 2 )n 20 (1,3)dioxane, where n is 2 to 5; W, Y and Z are independently selected from the group consisting of C, CRio and N, with the proviso that at least one of W, Y and Z must be C; Rio is selected from the group consisting of hydrogen and halogen; X is selected from the group consisting of 0, S, SO 2 , and NR 1 ; 25 Ru, is selected from the group consisting of hydrogen, lower alkyl, substituted lower alkyl, benzyl, substituted benzyl, phenyl, and substituted phenyl. The point of attachment of the W-X-Y-Z-C heterocyclic ring to the SO 2 group is not a limitation of the present invention. However, in one preferred embodiment, 12 WO 03/050062 PCT/US02/38117 the ring is attached to the SO 2 group through a carbon-atom. However, the ring may be attached through 0, S, or N heteroatoms. The compounds of formula (I) contain one or more asymmetric carbon atoms and some of the compounds may contain one or more asymmetric (chiral) centers and 5 may thus give rise to optical isomers and diastereomers. While shown without respect to stereochemistry in Formula (I), when the compounds of Formula (I) contain one or more chiral centers, at least the a-amino acid-derived chiral center must be of S stereochemistry. Most preferably, the carbon atom to which N, T, R 3 and R 4 are attached is of S-stereochemistry. 10 In one embodiment, the present invention is directed toward a process for preparing chiral S enantiomers of N-sulfonyl 2-aminoalcohols of the general formula Ia: R R
(CH
2 )n cl NH OH 0 0 15 wherein R is lower alkyl, substituted lower alkyl, lower alkenyl, substituted lower alkenyl, lower alkynyl, substituted lower alkynyl, cycloalkyl, substituted cycloalkyl, phenyl, substituted phenyl, benzyl, substituted benzyl, CH 2 cycloalkyl,
CH
2 -3-indole, CH(loweralkyl)-2-furan, CH(loweralkyl)-4-methoxyphenyl, CH(loweralkyl)phenyl, or CH(OH)-4-SCH 3 -phenyl and n is 0 to about 10. 20 Desirably, the compounds prepared according to the method of the invention contain at least one chiral carbon center, where R in the above-noted structures is the same. In certain desired embodiments, the R groups are methyl, ethyl, and n-propyl, and most preferably the R groups are ethyl. However, the invention further encompasses producing a-amino acids and 2-aminoalcohols of the general formulae 25 provided herein where the R groups are different. In these compounds one or more additional chiral centers may be present; however, the additional chiral centers must 13 WO 03/050062 PCT/US02/38117 be optically pure and must not interfere with the production of the chirally pure a amino acids, 2-aminoalcohols, and pure S enantiomers of N-sulfonyl 2-aminoalcohols of the present invention. In another preferred embodiment, the chiral carbon center is of S 5 stereochemistry which gives rise to enantiomerically pure products. In one embodiment, the method of the invention is used to produce chirally pure a-amino acids which are readily converted to the N-sulfonyl a-amino acids. For example, a chirally pure a-amino acid prepared according to the invention can be used to prepare a compound of Formula (I). Particularly desirable compounds of Formula 10 (I) include thiophenesulfonamides, and more desirably, 5-halo thiophenesulfonamides, and most desirably, 5-halo thiophene sulfonamides with p branches in the side chain of a primary alcohol. Thus, with respect to Formula (I), the compound produced by the invention desirably has a structure in which X is S, W is C, Y is C (or CRio) and Z is C (or CRio), and the sulfonamide is attached to C2 of the 15 thiophene ring. More desirably, X is S, W is C, Y is C (or CRio), Z is C (or CRio) and
R
6 is a halogen. Most desirably, X is S, W is C, Y is C, Z is C, R 6 is a halogen, T is C(OH)RiR 2 , where R 1 and R 2 are hydrogen, R3 is H, R 4 is a lower alkyl of S stereo chemistry, and R5 is H. Other desirable compounds of Formula (I) are furansulfonamides, in which X is 0, W is C, Y is C, and Z is C. In one particularly 20 desirable embodiment, the furansulfonamides of Formula (I) are further characterized by p-branches in the side chain of a primary alcohol. Thus, with respect to Formula (I), in these compounds T is C(OH)R 1
R
2 , in which R 1 and R2 are hydrogen, R3 is H,
R
4 is a lower alkyl of S-stereochemistry, R5 is H and R 6 is halogen. In preliminary screening assays in vitro and in vivo, selected compounds of these structures have 25 been found to have unexpectedly good beta-amyloid inhibitory activity, and in many cases, better activity than compounds of Formula (I) having other heterocycles (e.g., furans, where X is 0). However, other such compounds of Formula (I) are also useful for the purposes described herein. Additionally, other chirally pure a-amino acids and N-sulfonyl a-amino acids 30 prepared by the invention can be converted to the desired N-sulfonyl 2 aminoalcohols, which include the compounds of Formula (I). The compounds of Formula (I) are characterized by being sulfonamides of Formula (I), which have p 14 WO 03/050062 PCT/US02/38117 branches in the side chain of the primary alcohol group. Thus, with respect to Formula (I), in these compounds T is C(OH)RiR 2 , R 1 and R2 are hydrogen, R3 is H,
R
1 4 is a lower alkyl of S-stereochemistry, and R5 is H. These and chirally pure N sulfonyl a-amino acids can be prepared following the methods described herein. 5 A first method of preparation consists of reaction of a 2-aminoalcohol II with the appropriate sulfonyl halide in the presence of a base such as triethylamine (TEA) and in a suitable solvent to afford compounds of Formula III. For compounds where R2 and R1 are hydrogen, oxidation of the N-sulfonyl primary alcohol with pyridinium chlorochromate (PCC) or under Swern conditions then affords the corresponding 10 aldehyde IV which can be reacted with Grignard reagents (RMgX, where R is an organic radical and X is a halogen) to afford the secondary alcohols V as a mixture of diastereomers which can be separated by high performance liquid chromatography (HPLC) (Scheme 2). Scheme 2 0 OH s C O R5 OH
R
2
R
6 W y~ I
H
2 N X 0 ___N
R
4 R TEA R 4 3 X O R PCC or Swern when in III, RI=R 2 =H Y 0 R 5 OH H RYgX 0 R 5 0 R / R RMgX RH x O R 4
R
3 X R4 V IV 15 A second method of preparation involves reaction of an o-amino acid or ester IX with the appropriate sulfonyl halide in the presence of a base such as triethylamine and in a suitable solvent to afford compounds of Formula X (Scheme 3). The intermediate N-sulfonyl acid X (Rx=H) can be converted to the corresponding 15 WO 03/050062 PCT/US02/38117 primary alcohol VIII (R 1
=R
2 =H) utilizing standard methodology such as LiAlH 4 ,
B
2
H
6 or cyanuric chloride/NaBH 4 . The intermediate N-sulfonyl ester X (Rx=alkyl, Bn) can also be reduced to the corresponding primary alcohol VIII (R1=R 2 =H) utilizing standard methodology such as LiAlH 4 . Alternatively, the intermediate N 5 sulfonyl ester X (Rx=alkyl, Bn) can be converted to the aldehyde IV with DiBAL. Scheme 3 O R5 O R6 W H x O R R3 IV DiBAL Rx= alkyl, Bn 0 Y.---.-Z
O
O cl R O ORx TEA R6 ORx
R
4 RT x R 4
R
3 Rx=H or alkyl, Bn x IX RMgX (R 1
=R
2 ) LiAIH 4 or for Rx=H, Rx= alkyl, Bn Rx=H or alkyl, Bn B 2
H
6 0 R, OH 0 R 5 OH Y- --ZR2 Y Z | H NN R6 W R4 R, R6 W H x O 4 R x O 4 R 3 III VIII Finally, the intermediate N-sulfonyl ester X (Rx=alkyl, Bn) can be reacted with 2 equivalents of Grignard reagent to afford the tertiary alcohols III with
RI=R
2 =alkyl. Alternatively, for tertiary alcohols III with R 1 not equal to R 2 , the 10 corresponding Weinreb amide (see Scheme 11) of the N-sulfonyl acid can be prepared and subsequently reacted with R 1 MgX and R 2 MgX. For compounds of formula X (Rx=H) that have an asymmetric center at the a-amino acid carbon, the pure enantiomers can be obtained by standard resolution procedures employing recrystallization of salts formed with various chiral bases. 16 WO 03/050062 PCT/US02/38117 In a variation of the second method to prepare the primary alcohols, an a amino acid or ester (or N-protected derivative thereof) VI is first converted to the corresponding primary 2-aminoalcohol VII (using the methodology outlined in the previous paragraph), which is subsequently, after deprotection (if necessary), reacted 5 with the appropriate sulfonyl halide (Scheme 4) to afford compounds of Formula VIII. Scheme 4 P OH P 0 1. LiA1H 4 H ORx for Rx=H or alkyl, Bn H 4
R
4
R
3 or for Rx=H cyanuric Cl/NaBH 4 VII VI or B 2
H
6 P= H or protecting group 2. Deprotect _ C_ l 3. TEA R 6 W O R 5 OH Y I H SN R6 w H X O R 4
R
3 VIII For preparation of compounds derived from unnatural a-amino acids 10 containing beta branching in the amino acid side chain, a method of preparation based on the work of Hruby (Tet. Lett. 38: 5135-5138 (1997)) is outlined in Scheme 5. This route entails formation of the a,P-unsaturated amide XII of the Evans chiral auxiliary from an a, p-unsaturated acid XI, followed by conjugate addition of an organocuprate, trapping of the resulting enolate anion XIII with N-bromosuccinimide 15 (NBS), displacement of the bromide XIV with azide anion (provided by tetramethylguanidinium azide (TMGA)) to afford XV, followed by reduction to the 2 aminoalcohol and subsequent sulfonylation to afford the target compound XVI. In Schemes 2 through 5, R 5 is H. 17 WO 03/050062 PCT/US02/38117 Scheme 5 Ra Rn 1. tBuCOCI/TEA 0 3. CuBrDMS, RbMgX 0 O2H 2. L O Ph Ph XI Ph 4.'NBS X']' Ra Ra Hetaryl N3 Bri, O=S0= Ra 6. LAH 5. TMGA 0 ' -R ' N 0 R5" H " 7. HetarylSO 2 CA HOee Ph Ph XVI Xv XIV For the preparation of N-alkylated sulfonamides VIII (R 5 =alkyl etc.), the sulfonamide ester XVII can be N-alkylated by either treatment with a suitable base 5 such as potassium carbonate followed by the alkylating agent R 5 X or by employing Mitsunobu conditions (R 5 OH/DEAD, TPP). LiBH 4 reduction of the N-alkylated sulfonamide ester affords the N-alkylated sulfonamide in the primary alcohol series VIII (Scheme 6). These primary alcohols VIII can be converted to the secondary alcohols V or aldehyde IV series by chemistry that has been outlined above. 10 Alternatively, the N-alkylated sulfonamide esters, or their corresponding Weinreb aides, can be treated with Grignard reagents to afford the N-alkylated tertiary alcohols III. 18 WO 03/050062 PCT/US02/38117 Scheme 6 0- H? 0 OR 5 OH RI 6- N 1. Base/R 5 X R H R, ; ORx )W Swj x 0 RX ' R 3 - "k1 or X O R 4
R
3 RO/DEAD/TPP Rx= alkyl or Bn 2. LiBH 4 XVII When the heterocycle attached to the sulfonamide in the above alcohols is thiophene, the corresponding sulfone derivative XIX may be obtained by oxidation of the 5 thiophene compound XVIII with MCPBA (Scheme 7). Scheme 7 O Re OHR20 R OH 2 R / ..- MCPBA R / O R.N R 2
R
4 XI S R3 s 0 R 4
R
3 XVIII XIX 10 An alternate preparation of sulfonamides derived from unnatural 2 aminoalcohols utilizes the Bucherer modification of the Strecker a-amino acid synthesis (Scheme 8). In this route, an aldehyde XX is reacted with cyanide anion and ammonium carbonate to afford the hydantoin XXI, which is hydrolyzed to the a amino acid XXII. This compound is then reduced to XXIII and sulfonylated to afford 15 the desired compounds of Formula XXIV. 19 WO 03/050062 PCT/US02/38117 Scheme 8 O Rb Rb 1.NaCN/(NH 4
)
2
C
3 N0 Rb 2. NaOH H H Ra 1HO Ra )--HN Ra Z 0
H
2 N 0 XXII XX XXI 3. LiAlH 4 Rb Rb 10" R 4. HetaryISO 2 CI R HO Ra :HO Ra 0 HN-S-Hetaryl H 2 N XXIII XXIV For sulfonamides derived from 2-aminoalcohols containing an N or 0 heteroatom in the side chain, a route has been devised starting from D-serine 5 (Scheme 9). In this route, D-serine XXV is first sulfonylated to XXVI and subsequently converted to the ketone XXVII, which is reductively aminated to the target compounds of Formula XXVIII. 20 WO 03/050062 PCT/US02/38117 Scheme 9 0
CO
2 H C-- CO 2 H
H
2 N H R/ R- --- S-HN H HZl TEA > 6 H OH x 0 OH XXV XXVI BuLi R 0 MgX Rc NHRd Rco Y z 0H RdN}{ 2
H
/ S-HN N a c)3BH R -- W S--HN H -, 1H > 6 IH x 0OH HOAc x UOH XXVIII XXVII For sulfonamides derived from 2-aminoalcohols in the secondary alcohol series with R 1 =H and R 2
=CF
3 (compound XXIX), a method of preparation has been 5 devised that is outlined in Scheme 10 starting from the aldehyde IV (prepared as in Scheme 2). Scheme 10 0 R 5 0 1. TMSCF 3 /CsF 0 R6 OH R61H or TBw
CF
3 x 0 R 4
R
3 X2o 4
R
3 IV XXIX As has been mentioned in the section concerning Scheme 2, the preparation of 10 sulfonamides derived from 2-aminoalcohols in the secondary alcohol series V results in the formation of a diastereomeric mixture. An alternate method of preparation of these compounds that results in the production of a pure diastereomer is outlined in 21 WO 03/050062 PCT/US02/38117 Scheme 11 for compounds derived from L-isoleucine. This method, which utilizes chemistry previously employed by Roux (Tetrahedron 50: 5345-5360 (1994)), consists of addition of Grignard reagents to the Weinreb amide XXX (derived from the requisite a-amino acid) followed by stereospecific reduction of the ketone XXXI 5 to afford a single diastereomeric N-protected 2-aminoalcohol XXXII. Deprotection of this compound followed by reaction with sulfonyl chlorides affords the pure diastereomeric sulfonamide secondary alcohols of Formula XXXIII. Scheme 11 H 0 H 0 BOO RMgX BOC R NOMe - - R NaBH 4 O H OH Y-z1 1. TFA I L, S_-NN R6e Ni R :-- BOC NR x 0 2. TEA YK0 ~~W-cl x 0 xxxiii xxxii 10 When the heterocycle attached to the sulfonamide in the above alcohols is thiophene, the corresponding 5-iodo and 5-fluoro-thiophene derivatives may be obtained by conversion of the 5-bromo-thiophene derivative XXXIV (obtained as in Scheme 2) to a 5-trialkyltin-thiophene intermediate XXXV which can be converted to either the 5-iodo-thiophene (XXXVII) by treatment with sodium iodide and 15 chloramine T or the 5-fluoro-thiophene analog (XXXVI) by treatment with SELECTFLUORTM (Aldrich Chemical Co) (Scheme 12). 22 WO 03/050062 PCT/US02/38117 Scheme 12 O R, OH 0 R 5 OH Br /N
(R
3 Sn) 2 /n RLA Br
R
3 Sn_ II S O R 4
R
3 Pd (PPh 3
)
4 S O R 4
R
3 XXXIV NaI/CNorame T cl (B =V N+ (BF 4 ") O R OH F 0 R 5 OH R RI R R 3 S 0 R 4
R
3 XXXVH XXXVI Sulfonamides derived from cyclohexylglycinol substituted by alkoxy and amino groups at the 4 position of the cyclohexane ring can be prepared according to the methods described herein (Scheme 13). This route entails initial hydrogenation of 5 4-L-hydroxyphenylglycine XXXVIII, followed by sulfonylation, reduction of the carboxylic acid with diborane and formation of the N,O-acetonide XXXIX. The 4 hydroxy acetonide XXXIX is then O-alkylated using sodium hydride and an alkylating agent such as an alkyl or benzyl bromide. This is followed by removal of the protecting group by treatment with aqueous acid to afford the 4-ether derivatives 10 of Formula XXXX. Alternatively, the 4-hydroxy acetonide XXXIX can be oxidized to the 4-ketone which can be reductively aminated and deprotected to afford the corresponding 4-amino analogs of Formula XXXXI. 23 WO 03/050062 PCT/US02/38117 Scheme 13 H 01. H 2 /Ra-Ni HN2. Y-Z10 0 *OH Re >t-N x fa "W x 0 3. B 2
H
6 4. DMP/TsOH OHQ OH xxxvMI 5.PGC I XI 6. RNH 2 /Na(OAC) 3 BH 5.NaHIRBr 7. aq HOAc 6. aq HOAc O- OH 0 OH / H H II_ 0 0 NHR OR xxxxiXXXX 24 WO 03/050062 PCT/US02/38117 Where desired, oximes XXXXXIV can be derived from the corresponding aldehydes IV by standard methodology as depicted in Scheme 14. Scheme 14 OH 0 fOR 5
R
6 W H NH 2 OH R 6 H x R R 3 x 0 R 3 5 IV xxxxx1v 10 EXAMPLES The following examples are provided to illustrate the production of representative compounds according to the method of the invention. One skilled in the art will appreciate that although specific reagents and conditions are outlined in the following examples, these reagents and conditions are not a limitation on the 15 present invention. Example 1 Method 1 20 5-Chloro-N-[(1S)-2-ethyl-1-formylbutyllthiophene-2-sulfonamide 0 0 N 0 H H CI 25 WO 03/050062 PCT/US02/38117 A. 5-(1 -Ethyl-propyl)-imidazolidine-2,4-dione Sodium cyanide (12.0 g, 244.8 mmol) and 2-ethylbutyraldehyde (10.0 mL, 81.3 mmol) were added to ammonium carbonate (25.4 g, 325.3 mmol) in H 2 0 (300 mL). Ethanol (300 mL) was added and salts precipitated. The reaction mixture 5 was heated to 90 C. After 1 h, the mixture became homogeneous and was stirred at 90 C for 18 h. After cooling to 25'C, about 500 mL of solvent was removed in vacuo. Concentrated HCl was added to acidify the mixture to pH 1-2 and a precipitate formed. It was filtered and the precipitate was recrystallized from EtOAc to afford 5 (1-ethyl-propyl)-imidazolidine-2,4-dione as a white solid (12.9 g, 93%). Mass 10 Spectrum (-ESI): 169 (M-H)~. B. N-[(5-Chloro-2-thienyl)sulfonyl]-3-ethylnorvaline 5-(1-Ethyl-propyl)-imidazolidine-2,4-dione (12.3 g, 72.3 mmol) was dissolved in a 150 mL solution of aqueous NaOH (11.6 g, 289.2 nimol). The solution was heated by microwave in a sealed vessel for 1 h. (Microwave conditions: 15 min 15 @ 100 % power, 150 C, 50 psi, then 5 min 0% power, then 15 min @ 100% power, 150 'C, 50 psi, then repeat sequence.) Water and ammonium hydroxide were removed from the reaction mixture in vacuo and the resulting crude amino acid and NaOH mixture was used in the next reaction without further purification. The crude amino acid and NaOH mixture was dissolved in 300 mL of 20 water. The mixture was cooled to 0 C in an ice bath. 5-Chlorotbiophene-2-sulfonyl chloride (17.3 g, 79.5 mmol) was dissolved in 100 mL of THF and added dropwise to the reaction mixture over 0.5 h. After 1 h the reaction mixture was allowed to warm gradually to 25 0 C and stirred for 16 h. THF was removed in vacuo and then the mixture was acidified to pH 1 with IN HCl. After about 15 min, a precipitate began 25 to crash out of the milky white solution. After 1 h, the mixture was cooled in a refrigerator for 1 h and then filtered. The precipitate was washed with 1 N HCI to provide N-[(5-chloro-2-thienyl)sulfonyl]-3-ethylnorvaline as a white solid (18.5 g, 78%). Mass Spectrum (-ESI): 325 (M-H)~. C. N-[(5-Chloro-2-thienyl)sulfonyl]-3-ethyl-L-norvaline 30 (+)-(1S,2R)-Ephedrine heniihydrate (16.7 g, 95.6 mmol) was added to a suspension ofN-[(5-chloro-2-thienyl)sulfonyl]-3-ethylnorvaline (31.2 g, 95.6 mmol) in 185 niL of EtOH. The mixture was heated slightly to dissolve solids and a 26 WO 03/050062 PCT/US02/38117 precipitate formed. After cooling at 5 C for 18 h the resulting suspension was filtered and the precipitate was washed with cold EtOH and EtOAc to give 27 % yield of the diastereomeric salt. The salt was recrystallized from boiling EtOAc (420 mL), then filtered off The resulting white solid was then dissolved in 300 mL of EtOAc and 5 300 mL of IN HCl. The layers were separated and the organic extract was washed with IN HCl (2 x 200 mL), dried (Na 2 S04), and concentrated to give N-[(5-chloro-2 thienyl)sulfonyl]-3-ethyl-L-norvaline as a white solid (5.6 g, 18 %). Chiral HPLC [chiralpak AD (25 x 0.46 cm), 8:2 hexane (0.1% TFA): isopropanol, L-isomer elutes at 9.6 min and D-isomer elutes at 13.1 min] indicated 96% chiral purity. 10 [a]D 2 1 = +44.5- (c = 1% SOLUTION, MeOH). Mass Spectrum (-ESI): 325 (M-H)~ Anal. Calc'd for Cn 1
H
16 ClN04S 2 : C, 40.55; H, 4.95; N, 4.30. Found: C, 40.30; H, 4.78; N, 4.16. D. 5-Chloro-N-[(IS)-2-ethyl-1-(hydroxymethyl)butyl-2 thiophenesulfonamide 15 To N-[(5-Chloro-2-thienyl)sulfonyl]-3-ethyl-L-norvaline (5.6 g, 17.2 mmol) in THF (150 mL) at 0 C was added a solution of 1 M borane tetrahydrofuran complex in THF (69 mL, 69 mmol) dropwise via addition funnel. After 15 min, the reaction mixture was warmed to 25"C and stirred for 18 h. It was then quenched with 90 mL of 10% AcOH in MeOH slowly. Volatiles were removed in vacuo. The 20 residue was then dissolved in EtOAc (300 mL), washed with saturated aqueous NaHCO 3 (3 x 200 mL), dried (Na 2 SO4), and concentrated to a white precipitate (5.1 g, 96%'yield, 96 % chiral purity). The precipitate was recrystallized with heptane/EtOAc, 4:1, to give optically pure 5-chloro-N-[(1S)-2-ethyl-1 (hydroxymethyl)butyl]-2-thiophenesulfonamide as white needles (4.4 g, 81% yield). 25 mp 113-114'C. [a]D 2 1 =+45 (c = 1% SOLUTION, DMSO). Mass Spectrum (-ESI): 310 (M-H). Anal. Calc'd for C 1
H
18 ClN0 3
S
2 : C, 42.37; H, 5.82; N, 4.49. Found: C, 42.37; H, 5.79; N, 4.38. E. 5-Chloro-N-[(iS)-2-ethyl-1-formylbutyl]thiophene-2-sulfonamide Pyridinium dichromate (2.4 g, 6.4 mmol) was added to a solution of 5 30 chloro-N-[(1S)-2-ethyl-1-(hydroxymethyl)butyl]-2-thiophenesulfonamide (0.5 g, 1.6 mmol) in CH 2 Cl 2 (20 mL). After 18 h, the reaction mixture was filtered through a plug of Celite. The filtrate was concentrated and the resulting residue was purified by 27 WO 03/050062 PCT/US02/38117 silica gel column chromatography (eluant: 1:4 EtOAc-hexane) to give 5-chloro-N [(1S)-2-ethyl-l-formylbutyl]thiophene-2-sulfonamide as a white solid (303 mg, 61%). [ID5 =+136.76* (c = 1% SOLUTION, CHC1 3 ). Mass Spectrum (-ESI): 308 (M-H)-. Anal. Calc'd for CniH 16 ClNO 3
S
2 : C, 42.64; H, 5.21; N, 4.52. Found: C, 42.57; H, 5 5.24; N, 4.52. Example 2 Method 2 5-Chloro-N-[(1S)-2-ethyl-1-formylbutyllthiophene-2-sulfonamide A. (2S)-3 -Ethyl-2- {[(IS)- 1 -phenylethyl] amino } pentanenitrile 10 To (S)-(-)- a-methylbenzylamine hydrochloride salt (1.2 g, 7.6 mmol) in 80 mL of 1:1 MeOH/H 2 0 was added potassium cyanide (0.5 g, 7.6 mmol) and 2 methylbutyraldehyde (0.94 mL, 7.6 mmol). A precipitate formed after 30 min. After 20 h, the suspension was filtered and washed with H20 to give (2S)-3-ethyl-2-{[(1S) l-phenylethyl]amino}pentanenitrile as a white powder (1.29 g, 74%). Mass Spectrum 15 (+ESI): 310 (M+H)*. Anal. Calc'd for C 15
H
22
N
2 : C, 78.21; H, 9.63; N, 12.16. Found: C, 77.90; H, 9.75; N, 12.32. B. 3-Ethyl-N 2 -[(1S)-1-phenylethyl]-L-norvalinamide To 25 mL of sulfuric acid at 0 C was added (2S)-3-ethyl-2-{[(1S)-1 phenylethyl]amino}pentanenitrile (2.7 g, 11.6 mmol) portionwise. The mixture was 20 warmed to 25'C. After 2 days, the reaction mixture was poured over about 100 g of crushed ice. Concentrated NH 4 0H was added to neutralize the acid. This mixture was extracted with EtOAc (3 x 100 mL), dried over Na 2 S04, filtered and concentrated to give 3-ethyl-N 2 -[(1S)-1-phenylethyl]-L-norvalinamide (2.6 g, 90%), which was used in the next step without purification. Mass Spectrum (+ESI): 249 (M+H)*. 25 Anal. Calc'd for C 15
H
2 4
N
2 0: C, 72.54; H, 9.74; N, 11.28. Found: C, 72.24; H, 10.04; N, 11.01 C. 3-Ethyl-L-norvalinamide A mixture of 3-ethyl-N 2 -[(1S)-1 -phenylethyl] -L-norvalinamide (2.6 g, 10.5 mmol) and 5% Pd/C (800 mhg) was shaken for 24 h in a Parr apparatus under 3 30 atm of H2. The mixture was filtered through a plug of Celite and the solvent was removed in vacuo to give 3-ethyl-L-norvalinanide as a white solid (1.4 g, 93 %), 28 WO 03/050062 PCT/US02/38117 which was used in the next reaction without further purification. Mass Spectrum (+ESI): 145 (M+H)*. D. N-[(5-Chloro-2-thienyl)sulfonyl]-3-ethyl-L-norvaline 3-Ethyl-L-norvalinamide (1.2 g, 4.8 mmol) was dissolved in conc. HCl 5 (10 mL) and heated to 100 C for 16 h. The reaction mixture was concentrated to a white solid consisting of the amino acid hydrochloride salt and one equivalent of NH4Cl, which was used in the next reaction without purification. Amino acid hydrochloride salt with 1 equivalent of NH4C (0.28 g, 1.19 mmol) was dissolved in 6 mL of H20 and then NaOH (0.24 g, 6.00 mmol) was 10 added. The solution was cooled to 0 C and then 5-chlorothiophene-2-sulfonyl chloride (0.29 g, 1.32 mmol) in 6 mL of THF was added dropwise. The mixture was warmed to 25'C. After 19 h, THF was removed in vacuo. The remaining solution was diluted with 10 mL of H20 and washed with EtOAc (2 x 10 mL). The solution was acidified with 1N HCl and a precipitate formed. This was filtered to give N-[(5 15 chloro-2-thienyl)sulfonyl]-3-ethyl-L-norvaline as a white solid (0.17 g, 44%). Chiral HPLC indicates that only the S enantiomer is present. 5-Chloro-N-[(1S)-2-ethyl-1-(hydroxymethyl)butyl]-2-thiophenesulfonamide and 5-chloro-N-[(1S)-2-ethyl-1-formylbutyl]thiophene-2-sulfonamide were then prepared from N-[(5-chloro-2-thienyl)sulfonyl]-3-ethyl-L-norvaline according to 20 method 1 of Example 1. Example 3 5-Chloro-N-[(lS)- 2 -ethyl-1-(hydroxymethyl)butyl]-2-thiophenesulfonamide Into a 3L 3-necked flask equipped with a nitrogen inlet tube, a mechanical 25 stirrer, and an addition funnel with a stopper was placed lithium borohydride (145 mL of a 2 M solution in THF, 0.29 mol). The solution was placed under nitrogen and cooled to 0"C. Chlorotrimethylsilane (73.8 mL, 0.58 mol) was added dropwise over a period of 30 min. The ice bath was removed and the resulting slurry was stirred at 25*C for 30 min. The reaction mixture was cooled to 0"C and the 3-ethyl-L-norvaline 30 (21.1 g, 0.145 mol), which was prepared according to US Provisional Patent Application No. 60/339,264, was added in portions as a solid over a period of 15 min. The reaction mixture was allowed to warm slowly to 25"C as the ice bath melted. 29 WO 03/050062 PCT/US02/38117 After 3 days at 25'C, the reaction mixture was cooled to 0 0 C, and methanol (217 mL) was carefully added over a period of 80 min. The solution was stirred at 25"C for an additional 40 min, then concentrated under reduced pressure in a water bath at 60*C. The resulting slurry was made basic with 20% sodium hydroxide (37.5 mL). Water 5 (37.5 mL) was added, and the entire aqueous layer was extracted with methylene chloride (300 mL), and dried (Na 2
SO
4 ). Concentration under reduced pressure gave 3-ethyl-L-norvalinol as an oil (17.3 g, 91%), which was used immediately or stored in the freezer overnight: Opt. Rot. {a25D= -3.7' (1% solution, DMSO); 1H NMR (DMSO-d 6 , 500 MHz): 8 4.38 (broad s, 1H), 3.35 (dd overlapping with a broad s at 10 8 3.32, J=4.5, 10.3 Hz, 3H), 3.14 (dd, J=7.9, 10.2 Hz, 1H), 2.63 (m, 1H), 1.45-1.05 (m, 5H), 0.82 and 0.81(two overlapping triplets, J=7.4 Hz, 6H); MS(+ESI): [M+H]*, 132 (60%). A mixture of 3-ethyl-L-norvalinol (34.1 g, 0.26 mol) and methylene chloride (700 mL) was placed under Argon, and cooled to 0 0 C. Triethylamine (36.2 mL, 0.26 15 mol) was added, followed by the dropwise addition of 5-chlorothiophene-2-sulfonyl chloride (56.4 g, 0.26 mol) in methylene chloride (400 iL). The reaction mixture was allowed to warm slowly to 25'C as the ice bath melted. After 3 days at 25 C, the reaction mixture was divided into two-0.6 L portions. Each portion was diluted with ethyl acetate (1 L), and washed three times with saturated potassium phosphate 20 monohydrate (200 mL), once with brine (200 iL), and dried (Na 2
SO
4 ). Concentration under reduced pressure gave a white solid (74.5 g, 92%). The product (87.98 g) from several runs were combined and recrystallized from hot heptane:ethyl acetate (4:1, 775 mL) to give the title compound as crystals (74.9 g, 85%): mp 115 117.6'C; Opt. Rot. [a]D 2 5 +10.81o (1% solution, MeOH); 1H NMR (DMSO-d', 500 25 MHz): 6 7.71 (d, J=8.1 Hz, 1H), 7.44 (d, J=4.1 Hz, 1H), 7.22 (d, J=4.1 Hz, 1H), 4.56 (t, J=5.2 Hz, OH), 3.31-3.15 (m, 3H), 1.40-1.15 (m, 4H), 1.07 (m, 1H), 0.79 and 0.76 (two overlapping triplets, J=7.3 Hz, 6H); "C NMR (DMSO-d, 100 MHz): 6 141.75, 133.73, 130.95, 127.60, 60.41, 56.89, 41.57, 21.31, 20.80, 11.79, 11.51; MS(-ESI): [M-H]-, 1 chlorine isotope pattern, 310 (100/o), 312 (30%); Anal. Calc. for 30 CuiH is C1N0 3
S
2 : C, 42.37, H, 5.82, N, 4.49. Found: C, 42.34, H, 5.65, N, 4.43. Chiral HPLC (Chiralpak AD, 25x0.46 cm, eluant 8:2 hexane/isopropanol containing 0.1% 30 WO 03/050062 PCT/US02/38117 TFA, flow rate 0.5 mL/min, UV detection at 254 nm, retention times for the S and R isomers are 10.95 min and 11.95 min, respectively) revealed an S/R ratio of 100.0:0.0. Example 4 5 5-Chloro-N-[(1S)-2-ethyl-1-(1-hydroxyethyl)butyl]thiophene-2-sulfonamide \0 S H OH CI A solution of methylmagnesium bromide (1.4 M, 7.0 mL, 9.7 mmol) in toluene/THF (75:25) was added to a 0 C solution of 5-chloro-N-[(1S)-2-ethyl-1 10 formylbutyl]thiophene-2-sulfonanide (Example 1 or 2, 1.0 g, 3.2 mmol) in THE (30 mL). The mixture was warmed to 25"C and after 2 h was quenched carefully with saturated aqueous ammonium chloride (25 mL). The mixture was extracted with EtOAc (3 x 25 mL). The organic extract was dried over Na 2 S04, filtered and concentrated to give a colorless oil. The product was purified by column 15 chromatography (Biotage), eluant: 1:4 EtOAc-hexane, to afford 5-chloro-N-[(1S)-2 ethyl-1-(1-hydroxyethyl)butyl]thiophene-2-sufonamide as a white solid (876 mg, 83%). The product is a diastereomeric mixture with a ratio of 3:7. mp 95-98 0 C. Anal. Calc'd for Ci 2
H
2 oClNO 3
S
2 : C, 44.23; H, 6.19; N, 4.30. Found: C, 44.25; H, 6.35; N, 4.29. Mass Spectrum (-ESI): 324 (M-H)~. 20 Example 5 - Assay Conditions A. Semi-preparative RP-HPLC conditions: Gilson Semi-Preparative HPLC system with Unipoint Software. 25 Column: Phenomenex C18 Luna 21.6 mm x 60 mm, 5 L Solvent A: Water (0.02% TFA buffer) Solvent B: Acetonitrile (0.02 % TFA buffer) Solvent Gradient: Time 0 : 10 % B; 2.5 min : 10 % B; 14 min: 90 % B. 31 WO 03/050062 PCT/US02/38117 Flow Rate: 22.5 mL/min The product peak was collected based on UV absorption and concentrated. B. Analytical LCMS conditions: 5 Hewlett Packard 1100 MSD with ChemStation Software Column: YMC ODS-AM 2.0 mm x 50 mm 5 p. column at 23'C; 3 [tL injection; Solvent A: Water (0.02% TFA buffer) Solvent B: Acetonitrile (0.02 % TFA buffer) 10 Gradient: Time 0: 95% A; 0.3 min: 95% A; 4.7 min: 10% A; 4.9 min: 95% A. Flow rate 1.5 mL/min; Detection: 254 nm DAD; API-ES Scanning Mode Positive 150-700; Fragmentor 70 mV. 15 C. Analytical LCMS conditions: ZMD (Waters) or Platform (Micromass) or LCZ (Micromass) Column: Zorbax SB-C8 Solvent: Acetonitrile+H 2 0 containing 0. 1%TFA or 0. 1%FA Gradient: 2.5min 15% Acetonitrile - 95% Acetonitrile 20 Flow rate 3ml\min Detection: ELSD detection (SEDEX 55) UV 253 detection (Schimadzu) Example 6 -Repressor Release Assay (RRA) 25 The compounds generated as described in Examples 1 through 4 were tested in the RRA in accordance with published techniques [Shuey, D.J., Sheiffele, P., Jones, D., Cockett, MI., and Quinet, E.M. (1999), "Repressor release: a useful tool for monitoring amyloid precursor protein (APP) proteolysis in mammalian cells", Society for Neuroscience Abstracts, Vol. 25, 29h Annual Meeting of Society for 30 Neuroscience, Miami Beach, Florida, October 23-28, 1999]. Briefly, this assay is performed as follows. 32 WO 03/050062 PCT/US02/38117 A. Cell Culture CHO-KI cells are cultured in whole DMEM media (DMEM -High Glucose with 10% fetal bovine serum, 1% Non-essential Amino Acids, and 1% Penicillin-Streptomycin) at 37 0 C with 5% C0 2 . Two million cells are plated into 10 5 cm dishes 24 hrs prior to transfection. Transient transfections are completed as recommended by Gibco BRL using their Lipofectamine Plus system. First, 6 [tg of pRSVO-luc and 6 pg of APP lac construct DNA are added to 460 p.L Opti-Mem transfection media and incubated with 30 pL Plus reagent for 15 minutes. Then, a lipid mixture of 40 kL 10 Lipofectamine reagent and 460 pL Opti-Mem transfection media is incubated with the DNA-Plus reagent mixture for 15 minutes. During the DNA-lipid incubation, the CHO-KI cells are washed once and covered in 5.0 mL DMEM media without Penicillin-Streptomycin. The DNA-lipid preparation is then layered onto these cells and incubated at 37'C overnight. 15 One and one half million transfected cells per well (100 piL total volume) are plated into sterile, opaque Packard 96-well Cultur-Plates in clear DMEM whole media (DMEM - without phenol red) and incubated at 370 C with 5% CO 2 for 3-5 hours. B. Compound Dilution 20 Compounds are diluted using two different protocols; one protocol is used for compounds supplied neat (weighed powder in vial) and the other protocol is used for compounds supplied in solution (20 mM in DMSO in 96-well plates). For both protocols, 25 mM Hepes and 25 mM Hepes/1% DMSO are prepared fresh to be used as diluent, The Hepes/DMSO is used as the diluent control on all experimental 25 plates. The following table depicts the steps for compound dilution (please note that the last step is the addition of compound to cells/media in tissue culture plate): 33 WO 03/050062 PCT/US02/38117 Concentration Dilution x mg compound (vial) Stock Solution 10 mg/mL diluted with 100% DMSO 20 LL stock solution Dilution 1 1 mg/mL 180 tL 25mM Hepes 60 pL Dilution 1 Dilution 2 200 4g/mL 240 IL 25 mM Hepes Dilution 3 11.3[tL Dilution 2 (in Cell Plate) 20 pg/mL (in 100 tL cells/well) Because some compounds arrive in 96-well format at 20 mM, the following represents the protocol for their dilution (note that an average molecular weight of 5 these compounds was used to calculate these dilutions and as above, the last step is the addition of compound to cells/media in tissue culture plate): Concentration Dilution Stock Solution (original conc.) - 20 mM Solution 6 [L stock solution Dilution 1 -200 ptg/mL 194 iL 25 mM Hepes 11.3pL Dilution 2 Dilution 2 (in Cell Plate) ~20 tg/mL (in 100 p.L cells/well) Once compounds are diluted, they are applied in duplicate on cells in tissue 10 culture plates (prepared above). Cells are incubated with compound at 370 C with 5%
CO
2 for an additional 36-48 hours. C. Assay Measurement Luciferase assays (LucLite reagent, Packard) are performed and are read on a Packard TopCount instrument. Media is removed from each 96-well plate 15 and replaced with 100 ptL PBS per well (with Mg 2 + and Ca 2 +). An equal volume (100 tL) of the LucLite lysis/substrate buffer is added to each well and the plates are sealed and mixed in the dark on a rotary shaker for 15-30 minutes at room temperature. Luciferase readings are then taken on the TopCount instrument. Measurements are expressed as relative light units (RLU) and are calculated and 20 analyzed in MS Excel as follows. 34 WO 03/050062 PCT/US02/38117 D. Analysis of data The results of the assay with respect to the compounds exemplified herein are provided in the following table. A compound is considered active in RRA if it leads to at least a 1.5 fold increase in luciferase activity at 20 pg/mL and is non 5 toxic, as determined by loss of signal ( 0.75 fold increase). Fold increase is the amount of luciferase activity (measured in relative light units) over diluent control. SEM represents the standard error of the mean for fold increase (not shown). All compounds tested were found to be non-toxic. 10 Table Exl# Conc APPI Fold Name (pg/mL) Increase 1-3 20 4.9 5-chloro-N-[(1S)-2-ethyl-1-formylbutyl]thiophene-2 sulfonamide 4 20 7.2 5-chloro-N-[(1S)-2-ethyl-1-(1 9.7 hydroxyethyl)butyl]thiophene-2-sulfonamide All publications cited in this specification are incorporated herein by reference. While the invention has been described with reference to a particularly preferred embodiment, it will be appreciated that modifications can be made without 15 departing from the spirit of the invention. Such modifications are intended to fall within the scope of the appended claims. 35
Claims (42)
1. A method for producing a chirally pure a-amino acid or a salt thereof comprising the steps of: 5 (a) reacting an aldehyde and a cyanide salt with a chiral a-methylbenzylamine or a salt thereof and filtering to provide product (a); (b) reacting a strong inorganic acid and product (a); (c) neutralizing the acid of the reaction of (b); (d) extracting product (b) from the neutralized acid; 10 (e) hydrogenating product (b) in the presence of a catalyst to provide product (c); and (f) hydrolyzing product (c) in a strong aqueous acid to provide a salt of a chirally pure a-amino acid. 15
2. The method of claim 1, further comprising the step of neutralizing the salt of chirally pure a-amino acid to provide the chirally pure a-amino acid.
3. The method according to claim 1 or 2, wherein the reacting step (a) is permitted to continue for 12 to 24 hours. 20
4. The method according to any one of claims 1 to 3, wherein the reacting step (a) results in a suspension containing precipitate and further comprises the steps of filtering the suspension and washing the filtrate with water to provide a powder. 25
5. The method according to any one of claims I to 4, wherein the cyanide salt is selected from among sodium cyanide and potassium cyanide.
6. The method according to any one of claims 1 to 5, wherein the strong inorganic acid is sulfuric acid. 30
7. The method according to claim 6, wherein the sulfuric acid is at 0*C when product (a) is added.
8. The method according to any one of claims 1 to 7, wherein a strong 5 inorganic base is used in neutralizing step (c) and is ammonium hydroxide.
9. The method according to any one of claims 1 to 8, wherein the extracting step utilizes ethyl acetate as an extractant, and wherein the step further comprises the steps of drying, filtering, and concentrating to provide product (b). 10
10. The method according to any one of claims 1 to 9, wherein in the hydrogenating step (e), the catalyst is palladium.
11. The method according to any one of claims 1 to 10, wherein in the 15 hydrogenating step (e), the mixture is pressurized to 3 atm.
12. The method according to any one of claims 1 to 12, wherein the hydrolyzing step (f) is performed at a temperature of 100*C. 20
13. The method according to any one of claims 1 to 12, wherein the hydrogenating step (e) further comprises the steps of filtering the mixture and removing solvent.
14. The method according to any one of claims 1 to 13, wherein the 25 hydrolyzing step (f) is performed over a period of 16 hours.
15. The method according to any one of claims 1 to 15, wherein the hydrolyzing step (f) further comprises the step of concentrating the resulting reaction mixture to provide a product consisting of the amino acid hydrochloride salt and one 30 equivalent of ammonium chloride.
16. The method according to claim 15, further comprising the steps of dissolving .the product consisting of the amino acid hydrochloride salt and one equivalent of ammonium chloride in water and adding sodium hydroxide to form a solution. 5
17. The method according to any one of claims 1 to 16, wherein the chiral a amino acid is of the formula (R) 2 CH(CH 2 )nCH(CO 2 H)NHR', where n is 0 to about 10 and R is selected from the group consisting of lower alkyl, substituted lower alkyl, lower alkenyl, substituted lower alkenyl, lower .alkynyl, substituted lower alkynyl, cycloalkyl, substituted cycloalkyl, phenyl, substituted phenyl, benzyl, substituted benzyl, 10 CH 2 cycloalkyl, CH 2 -3-indole, CH(loweralkyl)-2-furan, CH(loweralkyl)-4 methoxyphenyl, CH(loweralkyl)phenyl, and CH(OH)-4-SCH 3 -phenyl; and R' is selected from the group consisting of H, lower alkyl, substituted lower alkyl, lower alkenyl, substituted lower alkenyl, lower alkynyl, substituted lower alkynyl, cycloalkyl, substituted cycloalkyl, heterocycle, substituted heterocycle, phenyl, substituted phenyl, 15 benzyl, and substituted benzyl.
18. A method for preparing a chiral 2-aminoalcohol, said method comprising the steps of: preparing a chiral a-amino acid according to claim 1; 20 reducing the a-amino acid to the 2-aminoalcohol; and recrystallizing the 2-aminoalcohol to afford the chirally pure 2-aminoalcohol.
19. A method for resolving an N-sulfonyl a-amino acid having a B-branched alkyl substituent for use in preparing a chirally pure N-sulfonyl a-amino acid, comprising 25 the steps of: (a) forming a mixture of chirally pure ephedrine hemihydrate and an N sulfonyl a-amino acid in ethanol at a molar ratio of 1:1, wherein the N-sulfonyl a-amino acid is selected from the group consisting of N-sulfonyl B-ethylnorvaline, N sulfonylvaline, and N-sulfonyl B-n-propylnorleucine; 30 (b) heating the mixture of (a) to about 80'C to dissolve the solids; (c) cooling the mixture to allow formation of a precipitate; (d) filtering the precipitate to give a diastereomeric salt; (e) recrystallizing the diastereomeric salt; (f) dissolving the recrystallized salt in an organic solvent and strong aqueous 5 acid and separating the layers to obtain an organic extract; (g) washing the organic extract; and (h) drying and concentrating the organic extract to provide chirally pure N-sulfonyl a-amino acid. 10
20. The method according to claim 19, wherein the N-sulfonyl ethylnorvaline is N-[(5-chloro-2-thienyl)sulfonyl]-3-ethylnorvaline; and wherein in step (d), the precipitate is washed with ethanol and ethyl acetate to give a diastereomeric salt. 15
21. The method according to claim 19 or 20, wherein the cooling step is performed at 5 0 C for 18 hours.
22. The method according to any one of claims 19 to 21, wherein the cooling is permitted to occur at room temperature. 20
23. The method according to any one of claims 19 to 22, further comprising the step of, following the cooling step, filtering the suspension formed.
24. The method according to any one of claims 19 to 23, wherein the 25 recrystallizing step is performed in boiling ethyl acetate.
25. The method according to any one of claims 19 to 24, further comprising the step of filtering off the recrystallized salt. 30
26. The method according to any one of claims 19 to 25, wherein the organic extract is washed with HCl.
27. The method according to any one of claims 19 to 26, wherein the organic extract is dried with Na 2 SO 4 .
28. A method for preparing a chiral N-sulfonyl 2-aminoalcohol comprising the steps of preparing a chiral N-sulfonyl a-amino acid according to claim 19; reducing the N-sulfonyl a-amino acid to. the N-sulfonyl 2 5 aminoalcohol; and recrystallizing the N-sulfonyl 2-aminoalcohol to afford the chirally pure N-sulfonyl 2-aminoalcohol.
29. The method according to claim 28, wherein the N-sulfonyl a aminoalcohol is of Formula (I), or pharmaceutically acceptable salt thereof, wherein 1 0 Formula (I) has the structure: 0 R5 T RsN x 4 R 3 (I) wherein: R 1 and R 2 are independently selected from the group consisting of hydrogen, alkyl, substituted alkyl, CF 3 , alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, 1 5 cycloalkyl, substituted cycloalkyl, phenyl, substituted phenyl, and (CH 2 ).(1,3)dioxane, where n is 2 to 5; R 3 is selected from the group consisting of hydrogen, alkyl, and substituted alkyl; R 4 is selected from the group consisting of hydrogen, alkyl, substituted alkyl, 20 alkylcycloalkyl, substituted alkylcycloalkyl, phenyl(substituted)alkyl, alkylOH, substituted alkyIOH, alkylOBn, substituted alkylOBn, alkylpyridyl, substituted 40 WO 03/050062 PCT/US02/38117 alkylpyridyl, alkylfuranyl, substituted alkylfuranyl, CH(OH)phenyl, CH(OH)substituted phenyl, alkenyl, substituted alkenyl, cycloalkyl, substituted cycloalkyl, N-substituted-piperidinyl, piperidinyl, substituted piperidinyl, tetrahydrothiopyran, substituted tetrahydrothiopyran, 2-indane, substituted 2-indane, phenyl, substituted phenyl, alkylNHR 7 , and substituted alkylNHR 7 ; with the proviso that R 3 and R4 are not both hydrogen; R 7 is alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, benzyl, substituted benzyl, alkylOH, substituted alkylOH, alkylSRs, or substituted alkylSR 8 ; R 8 is alkyl, substituted alkyl, benzyl, or substituted benzyl; or R 3 and R4 may be joined to form a ring; R 5 is selected from the group consisting of hydrogen, lower alkyl, substituted lower alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, CH 2 cycloalkyl, substituted CH 2 cycloalkyl, benzyl, substituted benzyl, and CH 2 CH 2 QR9; Q is 0, NH or S; R 9 is lower alkyl, substituted lower alkyl, phenyl, or substituted phenyl; R 6 is selected from the group consisting of hydrogen, halogen and CF 3 ; T is selected from the group consisting of OH 0 H ,and OH W, Y and Z are independently selected from the group consisting of C, CRio and N; R 1 0 is selected from the group consisting of hydrogen and halogen, with the proviso that at least one of W, Y and Z must be C; X is selected from the group consisting of 0, S, SO 2 , and NR 11 ; R 1 1 is selected from the group consisting of hydrogen, lower alkyl, substituted lower alkyl, benzyl, substituted benzyl, phenyl, and substituted phenyl; provided that when the compound contains one or more chiral centers, at least the a-amino acid-derived chiral center must be of S-stereochemistry. 41
30. The method according to claim 29, wherein R 6 is halogen.
31. The method according to claim 29 or 30, wherein R 6 is chlorine or bromine. 5
32. The method according to any one of claims 29 to 31, wherein T is C(OH)RiR 2 and R, and R 2 are each hydrogen.
33. The method according to any one of claims 29 to 32, wherein W and Z are 10 both C.
34. The method according to any one of claims 29 to 33, wherein R 4 is lower alkyl of S-stereochemistry. 15
35. The method according to any one of claims 29 to 34, wherein X is S, W is C, Z is C, R 6 is halogen, R 4 is lower alkyl of S-stereochemistry, R 3 is hydrogen, R 5 is hydrogen, and R, and R 2 are each hydrogen.
36. The method according to any one of claims 29 to 35, wherein R 3 CR 4 is 20 cyclohexyl.
37. The method according to any one of claims 29 to 36, wherein R 3 CR 4 is piperidine or N-substituted piperidine. 25
38. The method according to any one of claims 29 to 37, wherein X is S, and W, Y and Z are independently C or CR 10 . 30 A')
39. A method for producing chirally pure a-amino acids comprising the steps of: (a) reacting an aldehyde and potassium cyanide with an a methylbenzylamine or hydrochloride salt thereof and filtering to provide product (a); (b) reacting sulphuric acid and product (a); 5 (c) neutralizing the acid from the reaction of step (b); (d) extracting product (c) from the neutralized acid; (e) hydrogenating product (d) in the presence of a catalyst; (f) mixing product (e) in hydrochloric acid to provide a salt of chirally pure a-amino acid; and l0 (g) neutralizing the salt of chirally pure a-amino acid to provide the chirally pure a-amino acid.
40. A compound when produced by a method according to any one of claims 1, 18, 19, 28 or 39. 15
41. A method according to any one of claims 1, 18, 19, 28 or 39 substantially as hereinbefore described with reference to any of the Examples.
42. A compound according to claim 40 substantially as hereinbefore described with 20 reference to any of the Examples. 43
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
AU2009251195A AU2009251195A1 (en) | 2001-12-11 | 2009-12-24 | Production of chirally pure alpha-amino acids and n-sulfonyl alpha-amino acids |
Applications Claiming Priority (5)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US10/014,304 US6610734B2 (en) | 2000-12-13 | 2001-12-11 | Heterocyclic sulfonamide inhibitors of beta amyloid production |
US10/014,304 | 2001-12-11 | ||
US10/166,896 US6657070B2 (en) | 2000-12-13 | 2002-06-11 | Production of chirally pure α-amino acids and N-sulfonyl α-amino acids |
US10/166,896 | 2002-06-11 | ||
PCT/US2002/038117 WO2003050062A2 (en) | 2001-12-11 | 2002-11-26 | PRODUCTION OF CHIRALLY PURE α-AMINO ACIDS ADN N-SULFONYL α-AMINO ACIDS |
Related Child Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
AU2009251195A Division AU2009251195A1 (en) | 2001-12-11 | 2009-12-24 | Production of chirally pure alpha-amino acids and n-sulfonyl alpha-amino acids |
Publications (3)
Publication Number | Publication Date |
---|---|
AU2002351170A1 AU2002351170A1 (en) | 2003-06-23 |
AU2002351170B2 AU2002351170B2 (en) | 2009-09-24 |
AU2002351170B8 true AU2002351170B8 (en) | 2009-10-08 |
Family
ID=46204648
Family Applications (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
AU2002351170A Ceased AU2002351170B8 (en) | 2001-12-11 | 2002-11-26 | Production of chirally pure alpha-amino acids and N-sulfonyl alpha-amino acids |
AU2009251195A Abandoned AU2009251195A1 (en) | 2001-12-11 | 2009-12-24 | Production of chirally pure alpha-amino acids and n-sulfonyl alpha-amino acids |
Family Applications After (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
AU2009251195A Abandoned AU2009251195A1 (en) | 2001-12-11 | 2009-12-24 | Production of chirally pure alpha-amino acids and n-sulfonyl alpha-amino acids |
Country Status (7)
Country | Link |
---|---|
EP (1) | EP1461332A4 (en) |
JP (1) | JP4615861B2 (en) |
AU (2) | AU2002351170B8 (en) |
CA (1) | CA2470111A1 (en) |
MX (1) | MXPA04005366A (en) |
TW (1) | TWI260316B (en) |
WO (1) | WO2003050062A2 (en) |
Families Citing this family (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
SI1341779T1 (en) | 2000-12-13 | 2006-12-31 | Wyeth Corp | Heterocyclic sulfonamide inhibitors of beta amyloid production |
CL2004000647A1 (en) | 2003-03-31 | 2005-02-04 | Wyeth Corp | COMPOUNDS DERIVED FROM SULFONAMIDE HETEROCICLES CONTAINING FLUORO AND TRIFLUORO RENT; PHARMACEUTICAL COMPOSITION; PHARMACEUTICAL KIT; PREPARATION PROCEDURE; AND ITS USE AS BETA AMILOID INHIBITORS TO TREAT ALZHEIMER, ANGIOPATIA AM |
JP2007517905A (en) | 2004-01-16 | 2007-07-05 | ワイス | Beta-amyloidogenic heterocyclic sulfonamide inhibitors containing azoles |
AU2007243490A1 (en) | 2006-04-21 | 2007-11-08 | Wyeth | Production of chirally pure amino alcohol intermediates, derivatives thereof, and uses thereof |
Family Cites Families (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
DE3802981A1 (en) * | 1988-02-02 | 1989-08-10 | Basf Ag | METHOD FOR REDUCING REDUCABLE CONNECTIONS |
US5264577A (en) * | 1992-04-22 | 1993-11-23 | Warner-Lambert Company | Cyclic amino acids and derivatives thereof |
US5968942A (en) * | 1992-08-25 | 1999-10-19 | G. D. Searle & Co. | α- and β-amino acid hydroxyethylamino sulfonamides useful as retroviral protease inhibitors |
DK0656887T3 (en) * | 1992-08-25 | 1999-07-05 | Searle & Co | Hydroxyethylaminosulfonamides for use as inhibitors of retroviral proteases |
US6191306B1 (en) * | 1999-08-03 | 2001-02-20 | Eastman Chemical Company | Process for the preparation of cyclopropylglycine |
-
2002
- 2002-11-26 JP JP2003551090A patent/JP4615861B2/en not_active Expired - Fee Related
- 2002-11-26 EP EP02786817A patent/EP1461332A4/en not_active Withdrawn
- 2002-11-26 CA CA002470111A patent/CA2470111A1/en not_active Abandoned
- 2002-11-26 MX MXPA04005366A patent/MXPA04005366A/en active IP Right Grant
- 2002-11-26 WO PCT/US2002/038117 patent/WO2003050062A2/en active Application Filing
- 2002-11-26 AU AU2002351170A patent/AU2002351170B8/en not_active Ceased
- 2002-12-10 TW TW091135621A patent/TWI260316B/en active
-
2009
- 2009-12-24 AU AU2009251195A patent/AU2009251195A1/en not_active Abandoned
Also Published As
Publication number | Publication date |
---|---|
TWI260316B (en) | 2006-08-21 |
MXPA04005366A (en) | 2004-09-27 |
WO2003050062A3 (en) | 2003-11-20 |
AU2009251195A1 (en) | 2010-01-21 |
CA2470111A1 (en) | 2003-06-19 |
EP1461332A2 (en) | 2004-09-29 |
WO2003050062A2 (en) | 2003-06-19 |
EP1461332A4 (en) | 2009-10-21 |
TW200306292A (en) | 2003-11-16 |
AU2002351170B2 (en) | 2009-09-24 |
JP2005511727A (en) | 2005-04-28 |
JP4615861B2 (en) | 2011-01-19 |
AU2002351170A1 (en) | 2003-06-23 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
US6610734B2 (en) | Heterocyclic sulfonamide inhibitors of beta amyloid production | |
AU2002245123A1 (en) | Heterocyclic sulfonamide inhibitors of beta amyloid production | |
US7858658B2 (en) | Fluoro- and trifluoroalkyl-containing heterocyclic sulfonamide inhibitors of beta amyloid production and derivatives thereof | |
US6657070B2 (en) | Production of chirally pure α-amino acids and N-sulfonyl α-amino acids | |
CA2649396A1 (en) | Production of chirally pure amino alcohol intermediates, derivatives thereof, and uses thereof | |
AU2009251195A1 (en) | Production of chirally pure alpha-amino acids and n-sulfonyl alpha-amino acids | |
OA10518A (en) | Pyrrolidinyl hydroxamic acid compounds and their production process | |
CA2693959A1 (en) | Inhibitors of beta amyloid production | |
AU2002359519B2 (en) | Process for the synthesis of chirally pure beta-amino-alcohols | |
JP5410677B2 (en) | Method for producing optically active aminopentane derivative, intermediate and method for producing the same |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
TH | Corrigenda |
Free format text: IN VOL 23, NO 37, PAGE(S) 10212 UNDER THE HEADING APPLICATIONS ACCEPTED -NAME INDEX UNDER THE NAME WYETH, APPLICATION NO. 2002351170, UNDER INID (54) CORRECT THE TITLE TO READ PRODUCTION OF CHIRALLY PURE ALPHA-AMINO ACIDS AND N-SULFONYL ALPHA-AMINO ACIDS |
|
FGA | Letters patent sealed or granted (standard patent) | ||
MK14 | Patent ceased section 143(a) (annual fees not paid) or expired |