HK1081441B

HK1081441B - Proline derivatives having affinity for the calcium channel alpha-2-delta subunit

Info

Publication number: HK1081441B
Application number: HK06101658.5A
Authority: HK
Inventors: James Rawson David
Original assignee: 美国辉瑞有限公司
Priority date: 2002-10-31
Filing date: 2003-10-22
Publication date: 2009-01-23

Description

Proline derivatives having affinity for the alpha-2-delta subunit of calcium channels

Technical Field

The present invention relates to proline derivatives useful as medicaments, to processes for their preparation, to pharmaceutical compositions containing them and to their use for the treatment of the diseases listed below.

Background

Gabapentin (Neurontin)) Is used for the treatment ofAnticonvulsants of epilepsy, and recently the potential ability to treat neurogenic pain has been shown. It is 1- (aminomethyl) -cyclohexylacetic acid of the formula:

gabapentin is one of a series of compounds of the formula:

wherein R is hydrogen or lower alkyl and n is 4, 5 or 6. These compounds are described in US-A-4024175 and its division US-A-4087544. Gabapentin is used in the treatment of a number of disorders, including pain and epilepsy.

Gabapentin and related compounds, such as pregabalin, may be referred to as α -2- δ ligands. An alpha-2-delta receptor ligand is any molecule that binds to any subtype of the human calcium channel alpha-2-delta subunit. The calcium channel α -2- δ subunit includes several subtypes, which have been described in the literature, e.g. n.s.gee, j.p.brown, v.u.disananayake, j.off ford, r.thurlow and g.n.woodruff, J-pool-CHEM 271 (10): 5768-76, 1996, (type 1);

gong, j.hang, w.kohler, z.li, and T-z.su, j.membr.biol.184 (1): 35-43, 2001, (types 2 and 3);

e.marais, n.klugbauer and f.hofmann, molpharmacol.59 (5): 1243-1248, 2001 (types 2 and 3); and

qin, s.yagel, m.l.mommplaisir, e.e.codd, and m.r.d' andrea.mo1.pharmacol.62 (3): 485 496, 2002, (type 4). They may also be referred to as GABA analogs.

International patent application nos. WO0230871 and WO0222568 describe compounds of type I and type 11 respectively,

it also has affinity for gabapentin binding sites and has similar physiological activity as gabapentin, especially for analgesia.

International patent application No. WO0119817 describes 3-pyrrolidino-3' -pyridyl ether compounds, which are useful for controlling neurotransmitter release.

International patent application No. WO0222575 describes benzamidine derivatives which are serine protease inhibitors.

Certain compounds comprised in the broadest formula of the present invention have been disclosed for use unrelated to the present invention, in particular compounds according to table 1:

disclosure of Invention

The present invention provides proline derivatives and pharmaceutically acceptable salts, solvates, polymorphs and prodrugs thereof, useful for the treatment of various diseases including epilepsy, faintness attacks, hypokinesia, craniopathy, neurodegenerative diseases, depression, anxiety, panic, pain, fibromyalgia, sleep disorders, osteoarthritis, rheumatoid arthritis, and neuropathic conditions. The compounds provided are also useful in the treatment of visceral pain, functional bowel disorders such as gastroesophageal reflux, dyspepsia, irritable bowel syndrome and functional abdominal pain syndrome, as well as inflammatory bowel disorders such as Crohn's disease, ileitis, and ulcerative colitis, and other types of visceral pain associated with dysmenorrhea, pelvic pain, cystitis and pancreatitis. They are also useful for the treatment of premenstrual syndrome.

Accordingly, the present invention provides the use of a compound of formula (I) or a pharmaceutically acceptable salt, solvate or prodrug thereof in medical therapy:

in the formula

Or X is O, S, NH or CH₂And Y is CH₂Or a direct bond, or Y is O, S or NH and X is CH₂(ii) a And is

R is 3-12 membered cycloalkyl, 4-12 membered heterocycloalkyl, aryl or heteroaryl, wherein any ring may be optionally substituted with one or more substituents independently selected from the group consisting of: halogen, hydroxy, cyano, nitro, amino, hydroxycarbonyl, hydroxy-, cyano-, nitro-, amino-, or carboxyl group,

C₁-C₆Alkyl radical, C₁-C₆Alkenyl radical, C₁-C₆Alkynyl, alkynyl,

C₁-C₆Alkoxy, hydroxy C₁-C₆Alkyl radical, C₁-C₆Alkoxy radical C₁-C₆Alkyl, perfluoro C₁-C₆Alkyl, perfluoro C₁-C₆Alkoxy radical,

C₁-C₆Alkylamino radical, di-C₁-C₆Alkylamino radical, amino radical C₁-C₆Alkyl radical, C₁-C₆Alkylamino radical C₁-C₆Alkyl, di-C₁-C₆Alkylamino radical C₁-C₆Alkyl, aryl, heteroaryl, and heteroaryl,

C₁-C₆Acyl radical, C₁-C₆Acyloxy, C₁-C₆Acyloxy C₁-C₆Alkyl radical, C₁-C₆An amido group,

C₁-C₆Alkylthio radical, C₁-C₆Alkylthio carbonyl group, C₁-C₆Alkylthio radical, C₁-C₆An alkoxycarbonyl group, a carbonyl group,

C₁-C₆alkylsulfonyl radical, C₁-C₆Alkylsulfonylamino, aminosulfonyl, C₁-C₆Alkylaminosulfonyl, di-C₁-C₆An alkylaminosulfonyl group,

3-8 membered cycloalkyl, 4-8 membered heterocycloalkyl, phenyl and monocyclic heteroaryl.

Another aspect of the present invention provides the use of a compound of formula (1), or a pharmaceutically acceptable salt, solvate or prodrug thereof, in the manufacture of a medicament for the treatment of a disease in which the α -2- δ receptor is involved. Suitably, the disease in which the alpha-2-delta receptor is involved is selected from epilepsy, faintness attacks, hypokinesia, craniopathy, neurodegenerative diseases, depression, anxiety, panic, pain, fibromyalgia, irritable bowel syndrome, sleep disorders, osteoarthritis, rheumatoid arthritis, neuropathological disorders, visceral pain, functional bowel disease, inflammatory bowel disease, pain associated with dysmenorrhea, pelvic pain, cystitis and pancreatitis.

As a further alternative of a further aspect of the invention there is provided a method of treatment of a disease involving the alpha-2-delta receptor in a mammal, including a human, comprising the effective administration of a compound of formula (I) or a pharmaceutically acceptable salt, solvate or prodrug thereof.

According to formula (I), suitably X is O, S, NH or CH₂And Y is CH₂Or a direct bond, or X is CH₂And Y is O. Preferably, -Y-X-is methylene, methyleneoxy, methylenethio, oxydimethylene, amino, thio or an oxygen linkage. Particularly preferably, -Y-X-is an oxy (oxy), methylene or oxydimethylene linkage.

According to formula (I), R is suitably heteroaryl, aryl, 4-8 membered heterocycloalkyl or 3-12 membered cycloalkyl, optionally substituted with one or more substituents independently selected from: halogen, C₁-C₆Alkyl radical, C₁-C₆Alkoxy radical, C₁-C₆Alkoxy radical C₁-C₆Alkyl, perfluoro C₁-C₆Alkyl, perfluoro C₁-C₆Alkoxy, cyano, amino C₁-C₆Alkyl, di-C₁-C₆Alkylamino radical C₁-C₆Alkyl and monocyclic heteroaryl. R is more suitably optionally substituted aryl, 4-8 membered heterocycloalkyl or 3-12 membered cycloalkyl. R is preferably optionally substituted phenyl, cyclohexyl, dihydro-benzofuranyl or isoquinolinyl. More preferably R is optionally substituted phenyl. Most preferably, R is phenyl substituted in the meta position, and optionally disubstituted.

According to formula (I), suitable optional substituents on R are preferably in the meta position, independently selected from hydroxy, (C)₁-C₆) Alkoxy or halogen, preferably methoxy, fluoro, chloro or bromo, more preferably fluoro or chloro.

Particularly preferred compounds of the invention include those wherein each variable substituent (varvabre) in formula (I) is selected from the appropriate groups for each variable substituent. More preferred compounds of the invention include those wherein each variable substituent in formula (I) is selected from preferred or more preferred groups for each variable substituent.

It will be appreciated that certain compounds represented by formula (I), including those specifically described herein, are novel and thus individually and collectively constitute a further aspect of the invention.

Preferred compounds of formula (I) are selected from:

(2S, 4S) -4- (benzylsulfanyl) -pyrrolidine-2-carboxylic acid;

(2S, 4S) -4- [ (4-chlorobenzyl) oxy ] -pyrrolidine-2-carboxylic acid;

(2S, 4S) -4- [ (4-bromophenylsulfanyl ] -pyrrolidine-2-carboxylic acid;

(2S, 4S) -4-thiophenyl-pyrrolidine-2-carboxylic acid;

(2S, 4S) -4- [ 2-fluorophenoxy ] -pyrrolidine-2-carboxylic acid;

(2S, 4S) -4- [ (4-chlorophenoxy ] -pyrrolidine-2-carboxylic acid;

(2S, 4S) -4- [ 2-isoquinolinyloxy ] -pyrrolidine-2-carboxylic acid;

(2S, 4S) -4- (3-chloro-phenoxy) -pyrrolidine-2-carboxylic acid;

(2S, 4S) -4- (benzyloxy) -pyrrolidine-2-carboxylic acid;

(2S, 4S) -4- (3-fluoro-benzyl) -pyrrolidine-2-carboxylic acid;

(2S, 4S) -4- (2, 3-difluoro-benzyl) -pyrrolidine-2-carboxylic acid;

(2S, 4S) -4- (2, 5-difluoro-benzyl) -pyrrolidine-2-carboxylic acid;

(2S, 4S) -4-cyclohexylmethyl-pyrrolidine-2-carboxylic acid;

(2S, 4S) -4- (3-methoxy-benzyl) -pyrrolidine-2-carboxylic acid;

(2S, 4S) -4- (3-fluoro-phenoxymethyl) -pyrrolidine-2-carboxylic acid;

(2S, 4S) -4- (3-chloro-phenoxymethyl) -pyrrolidine-2-carboxylic acid;

(2S, 4S) -4- (2, 3-dihydro-benzofuran-6-yloxy) -pyrrolidine-2-carboxylic acid;

(2S, 4S) -4- (3-chloro-phenylamino) -pyrrolidine-2-carboxylic acid;

(2S, 4S) -4- (2, 5-difluoro-phenoxymethyl) -pyrrolidine-2-carboxylic acid;

(2S, 4S) -4- (2, 3-difluoro-phenoxymethyl) -pyrrolidine-2-carboxylic acid; and

(2S, 4S) -4- (3-methoxy-phenoxymethyl) -pyrrolidine-2-carboxylic acid.

More preferred compounds of formula (I) are selected from:

(2S, 4S) -4- (3-chloro-phenoxy) -pyrrolidine-2-carboxylic acid;

(2S, 4S) -4- (3-fluoro-benzyl) -pyrrolidine-2-carboxylic acid;

(2S, 4S) -4- (2, 3-difluoro-benzyl) -pyrrolidine-2-carboxylic acid;

(2S, 4S) -4- (2, 5-difluoro-benzyl) -pyrrolidine-2-carboxylic acid;

(2S, 4S) -4-cyclohexylmethyl-pyrrolidine-2-carboxylic acid;

(2S, 4S) -4- (3-methoxy-benzyl) -pyrrolidine-2-carboxylic acid;

(2S, 4S) -4- (3-fluoro-phenoxymethyl) -pyrrolidine-2-carboxylic acid;

(2S, 4S) -4- (2, 5-difluoro-phenoxymethyl) -pyrrolidine-2-carboxylic acid;

(2S, 4S) -4- (2, 3-difluoro-phenoxymethyl) -pyrrolidine-2-carboxylic acid; and

(2S, 4S) -4- (3-methoxy-phenoxymethyl) -pyrrolidine-2-carboxylic acid.

Certain compounds within the scope of formula (I) have been disclosed for non-therapeutic use. Thus, as a further aspect, the present invention provides a compound of formula (I), or a pharmaceutically acceptable salt, solvate, polymorph or prodrug thereof, excluding any compound previously disclosed in the art for non-therapeutic use, particularly those described in table 1 above, namely (2S, 4S) -4- (benzyloxy) pyrrolidine-2-carboxylic acid, (2S, 4S) -4- (benzylthio) pyrrolidine-2-carboxylic acid, (2S, 4S) -4-phenoxypyrrolidine-2-carboxylic acid, (2S, 4S) -4- (2-naphthoxy) pyrrolidine-2-carboxylic acid, (2S, 4S) -4- [ (4-methoxybenzyl) thio ] pyrrolidine-2-carboxylic acid, a pharmaceutically acceptable salt, solvate, polymorph or prodrug thereof, (2S, 4S) -4- [ (4-methylphenyl) thio ] pyrrolidine-2-carboxylic acid, (2S, 4S) -4- (phenylthio) pyrrolidine-2-carboxylic acid, (2S, 4S) -4- (2-naphthylthio) pyrrolidine-2-carboxylic acid, (2S, 4S) -4-benzylpyrrolidine-2-carboxylic acid, (2S, 4S) -4- (3-methoxybenzyl) pyrrolidine-2-carboxylic acid, (2S, 4S) -4- [4- (trifluoromethyl) benzyl ] pyrrolidine-2-carboxylic acid, (2S, 4S) -4- [ (4-nitrobenzyl) oxy ] pyrrolidine-2-carboxylic acid, (2S, 4S) -4- [ (4-cyclohexylbenzyl) thio ] pyrrolidine-2-carboxylic acid, (2S, 4S) -4- (4-chlorobenzyl) pyrrolidine-2-carboxylic acid, (2S, 4S) -4- (4-fluorophenoxy) pyrrolidine-2-carboxylic acid, (2S, 4S) -4- (4-methylphenoxy) pyrrolidine-2-carboxylic acid, (2S, 4S) -4- (3-methylthiophenoxy) pyrrolidine-2-carboxylic acid, (2S, 4S) -4- (4-chlorophenoxy) pyrrolidine-2-carboxylic acid, (2S, 4S) -4- (4-methoxyphenoxy) pyrrolidine-2-carboxylic acid, and mixtures thereof, (2S, 4S) -4- (1-Naphthyloxy) pyrrolidine-2-carboxylic acid, (2S, 4S) -4- (4-chlorophenylthio) pyrrolidine-2-carboxylic acid, trifluoromethylphenylthio) pyrrolidine-2-carboxylic acid, (2S, 4S) -4- (4-fluorophenylthio) pyrrolidine-2-carboxylic acid, (2S, 4S) -4- (4-acetoxyphenylthio) pyrrolidine-2-carboxylic acid, (2S, 4S) -4- (4-chlorobenzyloxy) pyrrolidine-2-carboxylic acid, (2S, 4S) -4- (4-phenyl-phenoxy) pyrrolidine-2-carboxylic acid, (2S, 4S) -4- (4-phenyl-phenylthio) pyrrolidine-2-carboxylic acid (2S, 4S) -4- (4-methyl-benzyloxy) pyrrolidine-2-carboxylic acid and (2S, 4S) -4- (4-fluorobenzyl) pyrrolidine-2-carboxylic acid.

In another aspect, the invention provides a compound of formula (la), (lb) or (lc):

in the formula R^aAnd R^bIndependently selected from hydrogen, halogen, hydroxyl and (C)₁-C₆) Alkoxycyano, nitro, amino, hydroxycarbonyl, nitro, amino, carboxyl, amino,

C₁-C₆alkyl radical, C₁-C₆Alkenyl radical, C₁-C₆Alkynyl, alkynyl,

3-8 membered cycloalkyl, 4-8 membered heterocycloalkyl, phenyl and monocyclic heteroaryl;

or a pharmaceutically acceptable salt, solvate or prodrug thereof in medical therapy.

Provided that

For the compounds of formulae (la) and (lb), R^aAnd R^bCannot all be hydrogen, and when R is^bWhen it is a para-substituent, R^aIt cannot be hydrogen and the compound can not be,

for compounds of formula (la), when R^aWhen it is methyl sulfide, R^bCannot be hydrogen, and

for the compound of formula (lb), when R is^aWhen it is methoxy, R^bAnd cannot be hydrogen.

For formulae (la), (lb) or (lc), R^aSuitably not hydrogen.

In the above definitions, halogen means fluorine, chlorine, bromine or iodine. Alkyl and alkoxy groups containing the desired number of carbon atoms may be unbranched or branched chains unless otherwise indicated. Examples of alkyl groups include straight and branched chain groups such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl and tert-butyl. Examples of alkoxy groups include straight and branched chain groups such as methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy and tert-butoxy. Alkenyl and alkynyl groups referred to herein include straight and branched chain cycloaliphatic groups having 1 double or triple bond, respectively. Examples of alkenyl and alkynyl groups include vinyl, prop-1-enyl (prop-1-enyl), prop-2-enyl and ethynyl, prop-1-ynyl and prop-2-ynyl, respectively.

As used herein, 4-8 membered heterocycloalkyl refers to a separately saturated or partially unsaturated ring system containing at least one ring heteroatom independently selected from O, S and N. As used herein, 4-12 membered heterocycloalkyl refers to a separately saturated or partially unsaturated ring or fused ring system containing at least one ring heteroatom independently selected from O, S and N. Polycyclic fused ring systems containing one or more carbocyclic fused saturated, partially unsaturated, or aromatic rings are therefore also within the definition of 4-12 membered heterocycloalkyl as long as the ring system also contains at least one fused ring containing at least one of the aforementioned heteroatoms. Suitable heterocycloalkyl groups include pyrrolidinyl, tetrahydrofuryl, dihydrofuranyl, tetrahydropyranyl, pyranyl, thiopyranyl, acridinyl, oxiranyl, methylenedioxy, benzopyranyl, isoxazolidin, 1, 3-oxazolidin-3-yl, isothiazolidinyl, 1, 3-thiazolidin-3-yl, 1, 2-pyrazolidin-2-yl, 1, 3-pyrazolidin-1-yl, piperidinyl, thiomorpholinyl, 1, 2-tetrahydrothiazin-2-yl, 1, 3-tetrahydrothiazin-3-yl, tetrahydrothiadiazinyl, morpholinyl, 1, 2-tetrahydrothiadiazin-2-yl, 1, 3-tetrahydrodiazin-1-yl, tetrahydroazepine, piperazinyl, chromanyl, 2, 3-dihydrobenzofuranyl, and the like.

Heteroaryl, as used herein, refers to a single aromatic ring or a fused aromatic ring system (preferably bicyclic) containing at least one ring heteroatom independently selected from O, S and N. Polycyclic fused rings containing one or more carbocyclic fused saturated, partially unsaturated, or aromatic rings are also within the definition of heteroaryl, provided that the ring system contains at least one fused aromatic ring, which ring contains at least one heteroatom as described above. Suitable heteroaryl groups include furyl, thienyl, thiazolyl, pyrazolyl, isothiazolyl, oxazolyl, isoxazolyl, pyrrolyl, triazolyl, tetrazolyl, imidazolyl, 1, 3, 5-oxadiazolyl, 1, 2, 4-oxadiazolyl, 1, 2, 3-oxadiazolyl, 1, 3, 5-thiadiazolyl, 1, 2, 3-thiadiazolyl, 1, 2, 4-thiadiazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, 1, 2, 4-triazinyl, 1, 2, 3-triazinyl, 1, 3, 5-triazinyl, pyrazolo [3, 4-b ] pyridyl, o-diazonaphthyl, pteridinyl, purinyl, 6, 7-dihydro-5H- [1] azoindenyl (pyridinyl), benzo [ b ] thiophenyl, 5, 6, 7, 8-tetrahydro-quinolin-3-yl, pyrrolyl, triazolyl, tetrazolyl, imidazolyl, 1, 3-thiadiazolyl, pyridazinyl, pyrimidinyl, pyrazinyl, pyridazinyl, triazinyl, 1, 2, 4-triazinyl, benzoxazolyl, benzothiazolyl, benzisothiazolyl, benzisoxazolyl, benzimidazolyl, thioindenyl, isothioindenyl, benzofuranyl, isobenzofuranyl, isoindolyl, indolyl, indolizinyl, indazolyl, isoquinolyl, quinolyl, phthalazinyl, quinoxalinyl, quinazolinyl, benzoxazinyl, and the like.

As used herein C₃-C₈Cycloalkyl refers to a single saturated or partially unsaturated carbocyclic ring system. As used herein C₃-C₁₂Cycloalkyl refers to a single or fused carbocyclic ring system containing at least one saturated or partially unsaturated ring, while the other rings in the fused system may be phenyl. Suitable cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, indane and 1, 2, 3, 4-tetrahydronaphthylene.

Aryl when used herein refers to phenyl or naphthyl.

Acyl, as used herein, refers to an aliphatic or cyclic hydrocarbon attached to a carbonyl group through which a substituent is attached.

Detailed Description

The compounds of the present invention may exist in unsolvated or solvated forms. The term "solvate" is used herein to describe a molecular complex comprising a compound of the invention and one or more pharmaceutically acceptable solvent molecules, such as ethanol. The term "hydrate" is used when the solvent is water.

Included within the scope of the present invention are complexes, such as cages, drug-host inclusion complexes (drug-host complexes) wherein the drug and host are present in stoichiometric or non-stoichiometric amounts, as opposed to the solvates described previously. Also included are complexes of drugs containing two or more organic and/or inorganic components, which may be in stoichiometric or non-stoichiometric amounts. The resulting complex may be ionized, partially ionized, or non-ionized. For a review of these complexes, see HaleblianJ Pharm Sci (8 months 1975),64(8)，1269-1288。

although the stereochemistry at the pyrrolidine ring of formula (I) is fixed, certain compounds of formula (I) containing one or more asymmetric carbon atoms may exist in the form of two or more stereoisomers. When the compound of formula (I) contains an alkenyl or alkenylene group, geometric cis/trans (or Z/E) isomers are possible. When the compound contains, for example, a keto or oxime group or an aromatic moiety, tautomerism (tautomerism) may occur. Which results in a single compound that may exhibit more than one type of isomerization.

All stereoisomers, geometric isomers, tautomers of the compounds of formula (I) are included within the scope of the present invention, including compounds that exhibit more than one type of isomerism, as well as mixtures of one or more thereof. Also included are acid adducts or base salts in which the counter ion is optically active, such as D-lactate or L-lysine; or racemic, e.g., DL-tartrate or DL-arginine.

The cis/trans isomers may be separated by conventional techniques well known to those of ordinary skill in the art, such as by chromatography and fractional crystallization.

Conventional techniques for the preparation/separation of the individual enantiomers include chiral synthesis from suitable optically pure precursors or separation of the racemates (or racemates of salts or derivatives) using, for example, chiral High Pressure Liquid Chromatography (HPLC).

Alternatively, the racemate (or a racemic precursor) may be reacted with a suitable optically active compound, for example an alcohol, or, when the compound of formula (I) contains an acid or base moiety, an acid or base such as tartaric acid or 1-phenylethylamine. The resulting diastereomeric mixtures can be separated by chromatography and/or fractional crystallization and one or both of the diastereomers converted to the corresponding pure enantiomers in a manner well known to those of ordinary skill in the art.

The chiral compounds of the invention (and chiral precursors thereof) can be obtained in enantiomerically enriched form on asymmetric resins using chromatography, typically HPLC, with a mobile phase consisting of a hydrocarbon, typically heptane or hexane, containing 0-50%, typically 2-20% isopropanol, and 0-5% alkylamine, typically 0.1% diethylamine. The concentrate of the eluent provides a concentrated mixture.

Mixtures of stereoisomers can be isolated by conventional techniques well known to those of ordinary skill in the art, see, e.g., "stereospecificity of Organic Compounds", E L Eliel (Wiley, New York, 1994).

The invention also includes all suitable isotopic variations of the compounds of the invention or pharmaceutically acceptable salts thereof. Isotopic variations of a compound of the present invention or a pharmaceutically acceptable salt thereof are defined as those wherein at least one atom is substituted by an atom having the same atomic number but an atomic weight different from the atomic weight usually found in nature.

Examples of isotopes suitable for inclusion in the compounds of the invention and pharmaceutically acceptable salts thereof include isotopes of the following elements: hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine and chlorine, e.g. each²H、³H、¹³C、¹⁴C、¹⁵N、¹⁷O、¹⁸O、³¹P、³²P、³⁵S、¹⁸F and³⁶Cl。

certain isotopically-labeled compounds of formula (I) and pharmaceutically acceptable salts thereof, for example, those in which a radioactive isomer is incorporated, e.g.³H or¹⁴C, compounds useful in drug and/or substrate tissue distribution studies. The radioactive isotope tritium is particularly preferred in view of its ease of preparation and existing detection equipment, i.e.³H and carbon-14, i.e.¹⁴Isotope of C.

With heavier isotopes such as deuterium, i.e.²H substitution may result in certain therapeutic advantages, leading to higher metabolic stability, such as increased in vivo half-life or a reduction in required dose, and is therefore preferred in some circumstances.

Using positron-emitting isotopes, e.g.¹¹C、⁸F、¹⁵O and¹³n substitution can be used in Positron Emission Tomography (PET) studies to examine substrate receptor occupancy.

Isotopically-labelled compounds of formula (I) can generally be prepared by conventional techniques known to those skilled in the art or by processes analogous to those described in the accompanying examples and preparations using an appropriate isotopically-labelled reactant in place of the unlabelled reactant previously used.

According to the invention, pharmaceutically acceptable solvates include those in which the crystallization solvent may be isotopically substituted, e.g. D₂O、d₆-acetone, d₆-those of DMSO.

The compounds of the invention are amino acids. Because the amino acids are amphoteric, pharmaceutically compatible salts can be salts of suitable non-toxic inorganic or organic acids or bases. Suitable acid addition salts are the hydrochloride/chloride, hydrobromide/bromide, hydroiodide, sulphate, bisulphate, nitrate, phosphate, biphosphate, acetate, fumarate, aspartate, benzenesulfonate, bicarbonate/carbonate, camphorsulphonate, D and L-lactate, D and L-tartrate, edisylate (edisylate), methanesulphonate, malonate, orotate, glucoheptonate, methylsulphate, stearate, glucuronate, 2-naphthalenesulphonate, toluenesulphonate, hydroxybenzylbenzoate, nicotinate, isethionate, malate, maleate, citrate, gluconate, succinate, saccharinate, benzoate, ethanesulphonate (esylate), trifluoroacetate and pamoate.

Suitable base salts are formed from bases which form non-toxic salts. Examples thereof include sodium salt, potassium salt, aluminum salt, calcium salt, magnesium salt, zinc salt, choline salt, diethanolamine salt, ethanolamine salt, arginine salt, glycine salt, tromethamine salt, benzathine salt, lysine salt, meglumine salt and diethylamine salt. The compounds of the present invention may also be formed as zwitterions.

Suitable salts of the amino acid compounds for use in the present invention are the hydrochloride salts. For a review of suitable Salts, see Stahl and Wermuth, Handbook of Pharmaceutical Salts: properties, Selection, and Use, Wiley-VCH, Weinheim, Germany (2002).

Salts having quaternary ammonium ions can also be prepared using, for example, tetramethylammonium ions. It has been found that oral absorption of certain drugs can be enhanced by preparing "soft" quaternary salts. The quaternary salts are referred to as "soft" quaternary salts because of their ability to react with normal quaternary salts, e.g., R-N⁺(CH₃)₃Instead, it may release the active drug upon hydrolysis. "Soft" quaternary salts have useful physical properties compared to basic drugs or salts thereof. The aqueous solubility may be improved compared to other salts, such as the hydrochloride salt, but more importantly it is possible to improve the intestinal absorption of the drug. The improved absorption may be due to the fact that the "soft" quaternary salts have surfactant properties and are capable of forming micelles with bile acids and unionized ion pairs, which are capable of more effectively penetrating the intestinal epithelium. Upon absorption, the prodrug is rapidly hydrolyzed, releasing the active parent drug.

Pharmaceutically acceptable salts of the compounds of formula (I) may be prepared by mixing together a solution of the compound of formula (I) and the acid or base as required. The salt may be precipitated from the solution and collected by filtration, or may be recovered by evaporation of the solvent. The degree of ionization in the salt can vary from fully ionized to nearly unionized.

Hereinafter, all references to compounds of formula (I) include references to salts, solvates and complexes thereof, and to solvates and complexes of salts thereof.

The compounds of the invention include compounds of formula (I) as defined hereinbefore, polymorphs, prodrugs and isomers thereof (including optical, geometric and tautomeric isomers) as defined hereinafter and isotopically labeled compounds of formula (I).

As mentioned, the present invention includes all polymorphs of the compound of formula (I) as defined below.

Also included within the scope of the present invention are so-called "prodrugs" of the compounds of formula (I). Certain derivatives of the compounds of formula (I), which may themselves have little or no pharmaceutical activity, are therefore capable of being converted, for example by hydrolytic cleavage or oxidative metabolism, when administered on or in the body, into compounds of formula (I) having the desired activity. These derivatives are referred to as "prodrugs". Further information on the use of prodrugs can be found in the ` Pro-drugs as Novel Delivery Systems, Vol.14, ACSSymposium Series (T Higuchi and W Stella) and ` Bioreversible Carriers in drug Delivery ` Pergamon Press, 1987(ed.E B Roche, American pharmaceutical Association).

Prodrugs according to the invention may be prepared, for example, by substituting appropriate functionalities present in compounds of formula (I) with certain moieties known to those of ordinary skill in the art as "precursor moieties", as described in "Design of ester Pro-drugs to organic precursors compositions" K.Beaumont et al, Current Drug Metabolism, 2003 and "Design of Pro-drugs" H.Bundgaard (Elsevier) 1985. In addition, certain compounds of the present invention may act as prodrugs of other compounds of the present invention. All protected derivatives and prodrugs of the compounds of the invention are included within the scope of the invention.

Some examples of prodrugs of the invention include:

(i) esters of carboxylic acid functionality (-COOH) of compounds of formula (I), e.g. with (C)₁-C₆) Alkyl instead of hydrogen, or carboxamides thereof, e.g. with amino functionality (-NH)₂-NHR or NRR ', wherein R and R' are each independently (C)₁-C₆) Alkyl) in place of hydroxy;

(ii) amides of secondary amino functionality (NHR where R is not H) of compounds of formula (I), e.g. with (C)₁-C₆) Alkanoyl radicalInstead of hydrogen.

Other examples of substituents and examples of other prodrug types according to the foregoing examples can be found in the foregoing references, which are incorporated herein by reference.

Aminoacyl-glycolic acid and-lactate are known as prodrugs of amino acids (WermuthC.G., Chemistry and Industry, 1980: 433-435). The carbonyl group of the amino acid can be esterified by known methods. Prodrugs and soft Drugs are known in the art (Palomino E., Drugs of the future, 1990; 15 (4): 361-.

The invention also relates to the therapeutic use of the compounds of the invention as medicaments for treating or alleviating the symptoms of neurodegenerative diseases. Such neurodegenerative diseases include, for example, Alzheimer's disease, Huntington's disease, Parkinson's disease, and Amyotrophic Lateral Sclerosis (Amyotropic late Sclerosis). The present invention also includes the treatment of neurodegenerative diseases known as acute brain injury. These include, but are not limited to: stroke, head trauma and asphyxia. Stroke refers to cerebrovascular disease and may be referred to as cerebrovascular accident (CVA), and includes acute thrombotic stroke. Strokes include local and global ischemia (global ischemia). And also transient cerebral ischemic stroke and other cerebral vascular problems associated with cerebral ischemia. These vascular diseases can often occur during a definite carotid endarterectomy or other cerebral or vascular surgery of a patient, or during diagnosis of blood vessels including cerebral angiography, etc. Other accidents are head trauma, spinal cord trauma or injuries commonly resulting from systemic hypoxia (hypoxia), hypoxia, hypotension or similar injuries seen during joint replacement (embole), hyper fusion and hypoxia. The invention can be used in a range of accidents, for example during cardiac bypass surgery (cardiac bypass surgery), in accidents with intracranial haemorrhages, in perinatal asphyxia, in cardiac arrest and status epilepticus.

The skilled practitioner will be able to determine the appropriate situation in which the target is susceptible to or at risk of, for example, stroke and stroke resulting from administration of the methods of the invention.

The compounds of the invention are useful in the general treatment of pain, in particular neuropathic pain. Physiological pain is an important protective mechanism designed to warn of the risk of potentially harmful stimuli from the external environment. The system operates through a specific series of primary sensory neurons and is triggered by toxic stimuli through a peripheral switch mechanism (MILLAN 1999 prog. neurobio.57: 1-164 for a review of the whole). These sensory fibers, called nociceptors, are characterized by small diameter axons with small conduction velocities. Nociceptors encode the intensity, duration and amount of noxious stimuli, as well as the location of the stimuli, which is obtained as a result of their topographical organized injection into the spinal cord. Nociceptors are found on nociceptive nerve fibers. The fibers are of two main types, a- δ fibers (myelinated) and C fibers (non-myelinated). The activity produced by nociceptor input is processed in the dorsal horn and then transported to the ventral basal thalamus, either directly or via the brainstem relay nuclei, and then to the cerebral cortex, where a sensation of pain is produced.

Intense acute and chronic pain can involve the same pathways driven by pathophysiological processes and are halted as described to provide a protective mechanism and help alleviate symptoms associated with a wide range of disease states. Pain is a characteristic of many trauma and disease states. When substantial damage to body tissue occurs through disease or trauma, the nature of nociceptor activation is altered. There is sensitization around the site that partially surrounds the lesion and is centered at the site of termination of the nociceptors. This results in hypersensitivity in normal tissue at and near the site of the lesion. In acute pain, these mechanisms may be useful and take into account that a repair process takes place, and when the injury is cured, the hypersensitivity returns to normal. However, in many chronic pain states, allergies persist longer than the healing process and are often due to nervous system injury. Such damage often results in maladaptation of afferent fibers (Woolf & Salter 2000Science 288: 1765-. Clinical pain exists when there is discomfort and an abnormal sensitivity profile in the patient's symptoms. Patients tend to be very heterogeneous and are accompanied by various pain symptoms. There are some typical pain subtypes: 1) spontaneous pain, which may be dull, burning or stinging; 2) increased pain in response to noxious stimuli (hyperalgesia); 3) pain (allodynia) produced by normal, non-noxious stimulation (Meyer et al, 1994 Textbook of Pain 13-44). While patients with back pain, arthritic pain, CNS trauma, or neuropathic pain may have similar symptoms, the underlying mechanisms are different and thus may require different treatment strategies. Pain can therefore be divided into several distinct areas due to pathophysiology, including nociceptive, inflammatory, neuropathic pain, etc. It should be noted that some types of pain have multiple etiologies and can therefore be divided into more than one area, with back pain, cancer pain having both nociceptive and neuropathic components.

Nociceptive pain is caused by tissue injury or by intense stimuli with the potential to produce injury. Afferent pain can be activated by stimulation of conduction by nociceptors at the site of injury and sensitize the spinal cords at the level of their termination. This then relays to the painful brain on the spinal cord (Meyer et al, 1994 Textbook of Pain 13-44). Activation of the nociceptors activates two types of afferent nerve fibers. The myelinated a-delta fibers are transported rapidly and are responsible for the sensation of acuteness and tingling, whereas non-myelinated C fibers are transported at a slower rate and are less slow or sore to deliver. Moderate to severe acute nociceptive pain is a prominent feature from, but not limited to: strain/sprain, post-operative pain (pain after any type of surgical procedure), post-traumatic pain, burn, myocardial infarction, acute pancreatitis and renal colic. Moreover, cancer-related acute pain syndromes are often due to therapeutic interactions such as chemotherapy toxicity, immunotherapy, hormonal therapy and radiotherapy. Moderate to severe acute nociceptive pain is a prominent feature of cancer pain, but is not limited to this, and can be tumor-related pain (e.g., bone, headache and facial, visceral) or associated with cancer therapy (e.g., post-chemotherapy (postchemothermy) syndrome, chronic post-operative pain syndrome, post-radiation syndrome), back pain (which may be due to herniated or ruptured intervertebral or lumbar facet joint abnormalities, sacral joint abnormalities, paravertebral muscle abnormalities, or posterior longitudinal ligament abnormalities.

Neuropathic pain is defined as pain caused or produced by primary lesions or dysfunctions in the nervous system (IASP definition). Nerve damage can be caused by trauma and disease, and thus the term "neuropathic pain" encompasses a number of conditions of different etiology. These include, but are not limited to, diabetic neuropathy, postherpetic neuralgia, back pain, cancer neuropathy, HIV neuropathy, phantom limb pain, carpal tunnel syndrome, chronic alcoholism, hypothyroidism, trigeminal neuralgia, uremia, or vitamin deficiency. Neuropathic pain is pathological in that it has no protective effect. It usually exists well after the initial cause has disappeared, generally lasting several years, significantly reducing the quality of life of the patient (Woolf and Mannion 1999 Lancet 353: 1959-1964). Symptoms of neuropathic Pain are difficult to treat because they are usually heterogeneous even among patients with the same disease (Woolf & Decoster 1999 Pain Sup.6: S141-S147; Woolf and Mannion 1999 Lancet 353: 1959-1964). They include spontaneous pain, which may be continuous or episodic, and allodynia, such as hyperalgesia (increased sensitivity to noxious stimuli) and allodynia (sensitivity to normally innocuous stimuli).

The inflammatory process is a complex series of biochemical and cellular events that are triggered in response to tissue damage or the presence of impurities, which leads to swelling and Pain (Levine and Taiwo 1994: Textbook of Pain 45-56). Arthritic pain constitutes the majority of the overall inflammatory pain population. Rheumatoid disease is one of the most common chronic inflammatory states in developed countries, and rheumatoid arthritis is a common cause of disability. The exact etiology of RA is unknown, but current hypotheses suggest that genetic and microbiological factors may be important (Grennan & Jayson 1994 Textbook of Pain 397-407). It is estimated that about 1600 million Americans suffer from symptoms of Osteoarthritis (OA) or degenerative joint disease, most of which are over 60 years old, and that this is expected to exceed 4000 million as the population ages, making this a significant public health problem (Houge & Mersfelder 2002 Ann Pharmacother.36: 679-. Most patients with OA seek medical treatment due to pain. Arthritis has a great impact on psychosocial and somatic functions and is known to be a major cause of disability in the elderly. Other types of inflammatory pain include, but are not limited to, Inflammatory Bowel Disease (IBD), and other types of pain include, but are not limited to:

musculo-skeletal diseases include, but are not limited to, myalgia, fibromyalgia, spondylitis, seronegative (non-rheumatoid) arthropathy, non-articular rheumatism, dysphenopathgy, glycogenolysis, polymyositis, pyogenic myositis.

Central pain or "thalamic pain" as defined by pain caused by damage or dysfunction of the nervous system, including but not limited to central post-stroke pain, multiple sclerosis, spinal cord injury, parkinson's disease and epilepsy.

-cardiac and vascular pain including but not limited to angina, myocardial infarction, mitral stenosis, pericarditis, raynaud's disease, sclerodoma, skeletal muscle ischemia.

Visceral pain and gastrointestinal discomfort. Viscera includes organs in the abdominal cavity. These organs include the sexual organs, spleen and part of the digestive system. Visceral related pain can be divided into digestive visceral pain and non-digestive visceral pain. Commonly encountered Gastrointestinal (GI) diseases include Functional Bowel Disease (FBD) and Inflammatory Bowel Disease (IBD). These GI diseases include a wide range of disease states which are currently only moderately controlled, including-for FBD, gastroesophageal reflux, dyspepsia, Irritable Bowel Syndrome (IBS) and Functional Abdominal Pain Syndrome (FAPS), and-for IBD, crohn's disease, ileitis and ulcerative colitis, all of which regularly produce visceral pain. Other types of visceral pain include pain associated with dysmenorrhea, pelvic pain, cystitis and pancreatitis.

-headache, including but not limited to migraine, migraine aura, migraine without aura, cluster headache, tension headache.

Orofacial pain, including but not limited to dental pain, temporomandibular muscularis disci pain.

The compounds of the present invention are also expected to be useful in the treatment of depression. Of the causes, depression may be the source of an organic disease, secondary to stress associated with personal loss, or primary. Some forms of depression have a strong familial propensity to develop, suggesting a mechanical cause for at least some forms of depression (mechanistic house). The diagnosis of depression is primarily made by quantifying the changes in the mood of the patient. The assessment of these emotions is typically performed by a physician or neuropsychologist using an effective rating scale, such as the Hamilton depression rating scale or the Brief psychopath rating scale. Numerous other scales have been developed to quantify and measure the degree of mood changes in patients with depression, such as insomnia, inability to concentrate, lack of energy, feelings of worthlessness, and guilt. Criteria for diagnosing depression as well as all Psychiatric diagnoses are collected at the Diagnostic and Statistical Manual of Mental Disorders (fourth edition) called DSM-IV-R Manual, published by the American Psychiatric Association, 1994.

In another aspect, methods of treating a disease selected from the group consisting of: epilepsy, faintness attacks, hypokinesia, craniopathy, neurodegenerative diseases, depression, anxiety, panic, pain, irritable bowel syndrome, sleep disorders, osteoarthritis, rheumatoid arthritis, neuropathological diseases, visceral pain, functional bowel diseases, inflammatory bowel diseases, pain associated with dysmenorrhea, pelvic pain, cystitis and pancreatitis, comprising administering to a mammal in need of such treatment a therapeutically effective amount of a compound of formula (I).

The biological activity of the alpha-2-delta ligand of the present invention can be expressed by³H]Gabapentin and from pig brainThe α 2 δ subunit of tissue, measured in a radioligand binding assay (Gee N.S., Brown J.P., Dissanayake V.U.K., Offord J.S., Thurlow R., Woodruff G.N., J.Bio1.chem., 1996; 271: 5879-. The results can be expressed in terms of μ M or nM α 2 δ binding affinity.

The compounds of the invention may also be administered alone, simultaneously or sequentially in combination with one or more other pharmacologically active agents. Suitable agents are, in particular, agents for the treatment of pain, including:

i) opioid analgesics such as morphine, heroin, hydromorphone, oxymorphone, levorphanol, methadone, meperidine, fentanyl, codeine, dihydrocodeine, oxycodone, hydrocodone, propoxyphene, nalmefene, nalorphine, naloxone, naltrexone, buprenorphine, butorphanol, nalbuphine and pentazocine;

ii) opioid antagonists (opioid antagonists), such as naloxone, naltrexone;

iii) non-steroidal anti-inflammatory drugs (NSAIDS), such as aspirin, diclofenac, diflusinal, etodolac, fenbufen, fenoprofen, flufenisal, flurbiprofen, ibuprofen, indomethacin, ketoprofen, ketorolac, meclofenamic acid, mefenamic acid, nabumetone, naproxen, oxaprozin, phenylbutazone, piroxicam, sulindac, tolmetin, zomepirac and pharmaceutically acceptable salts thereof;

iv) barbiturate sedatives such as amobarbital, alprenol, sec-butyl barbital, butabital, mebendal, methamphetal, methohexital, pentobarbital, phenobarbital, secobarbital, talbarbital, theamylal, thiopental and pharmaceutically acceptable salts thereof;

v) benzodiazepines with sedative actionClass (iii) e.g. chlorine nitrogenChlorine nitrogenSalts (clorazepate), diazepam, flurazepam, lorazepam, oxazepam, temazepam, triazolam, and pharmaceutically acceptable salts thereof;

vi) H with sedative action₁Antagonists such as diphenhydramine, mepyramine, promethazine, chlorpheniramine, clocycline, and pharmaceutically acceptable salts thereof;

vii) hybrid sedatives (miscella neurous sedatives), such as glutethimide, toluate, methaqualone, chloraldoprine, and pharmaceutically acceptable salts thereof;

viii) skeletal muscle relaxants, such as baclofen, carisoprodol, chlorzoxazone, cyclobenzaprine, methocarbamol, orphradine and pharmaceutically acceptable salts thereof,

ix) NMDA receptor antagonists such as dextromethorphan ((+) -3-hydroxy-N-methyllevopyran) and its metabolite dextromethorphan ((+) -3-hydroxy-N-methyllevopyran), ketamine, memantine, pyrroloquinoline quinone and cis-4- (phosphonomethyl) -2-piperidinecarboxylic acid, and pharmaceutically acceptable salts thereof;

x) α -adrenergic active compounds, such as doxazosin, tamsulosin, clonidine and 4-amino-6, 7-dimethoxy-2- (5-methanesulfonamido-1, 2, 3, 4-tetrahydroisoquinol-2-yl) -5- (2-pyridyl) quinazoline;

xi) tricyclic antidepressants, such as desipramine, imipramine, amytriptiline and nortriptyline;

xii) anticonvulsants such as carbamazepine and valproate;

xiii) 5-hydroxytryptamine reuptake inhibitors such as fluoxetine, paroxetine, citalopram and sertraline;

xiv) mixed 5-hydroxytryptamine-norepinephrine reuptake inhibitors such as milnacipran, venlafaxine, and duloxetine;

xv) norepinephrine reuptake inhibitors, such as reboxetine;

xvi) tachykinin (NK) antagonists, in particular NK-3, NK-2 and NK-1 antagonists, such as (alpha R, 9R) -7- [3, 5-bis (trifluoromethyl) benzyl ] -8, 9, 10, 11-tetrahydro-9-methyl-5- (4-methylphenyl) -7H- [1, 4] diazocine [2, 1-g ] [1, 7] naphthyridine (naphthyridine) -6-13-dione (TAK-637), 5- [ [ (2R, 3S) -2- [ (1R) -1- [3, 5-bis (trifluoromethyl) phenyl ] ethoxy-3- (4-fluorophenyl) -4-morpholinyl ] methyl ] -1, 2-dihydro-3H-1, 2, 4-triazol-3-one (MK-869), lanopiptan (lanepitant), dapitant (dapitant), and 3- [ [ 2-methoxy-5- (trifluoromethoxy) phenyl ] methylamino ] -2-phenyl-piperidine (2S, 3S);

xvii) muscarinic antagonists such as oxybutynin (oxybutin), tolterodine, propiverine, tobramide chloride, and darifenacin;

xviii) COX-2 inhibitors, such as celecoxib, rofecoxib, and valdecoxib;

xix) non-selective COX inhibitors (preferably with GI protection), such as nitroflurbiprofen (HCT-1026);

xx) PDEV inhibitors, for example sildenafil, vardenafil (Bayer), tadalafil (tadalafil) (Icos Lilly), 1- { 6-ethoxy-5- [ 3-ethyl-6, 7-dihydro-2- (2-methoxyethyl) -7-oxo-2H-pyrazolo [4, 3-d ] pyrimidin-5-yl ] -3-pyridylsulfonyl } -4-ethylpiperazine, 5- (5-acetyl-2-butoxy-3-pyridyl) -3-ethyl-2- (1-ethyl-3-azetidinyl) -2, 6-dihydro-7H-pyrazolo [4, 3-d ] -pyrimidin-7-one and 5- [ 2-ethoxy-5- (4-ethylpiperazin-1-ylsulfonyl) pyridin-3-yl ] -3-ethyl-2- [ 2-methoxyethyl ] -2, 6-dihydro-7H-pyrazolo [4, 3-d ] pyrimidin-7-one;

xxi) coal tar analgesics (coal-tar analgesics), particularly acetaminophen;

xxii) neuroleptic agents, such as droperidol;

xxiii) Vanilloid receptor agonists, such as anti-urinary-loss-of-care drugs (resimultifaxin);

xxiv) β -adrenergic compounds, such as propranolol;

xxv) local anesthetics, such as mexiletine;

xxvi) corticosteroids, such as dexamethasone;

xxvii) 5-hydroxytryptamine receptor agonists and antagonists;

xxviii) cholinergic (nicotinic) analgesics; and

xxix) admixtures (miscella neous agents) such as Tramadol

Accordingly, the present invention also provides a combination drug (combination) comprising a compound of the above formula (I) or a pharmaceutically acceptable salt, solvate or prodrug thereof, and a compound or compounds selected from the classes (I) to (XXIX). Also provided is a pharmaceutical composition comprising the combination together with a pharmaceutically acceptable excipient, diluent or carrier, particularly for use in the treatment of a disease involving an alpha-2-delta ligand.

Thus, in a further aspect, the present invention provides a combination pharmaceutical product comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof and a PDEV inhibitor. Preferred PDEV inhibitors are selected from sildenafil, vardenafil, tadalafil, 1- { 6-ethoxy-5- [ 3-ethyl-6, 7-dihydro-2- (2-methoxyethyl) -7-oxo-2H-pyrazolo [4, 3-d ] pyrimidin-5-yl ] -3-pyridylsulfonyl) -4-ethylpiperazine, 5- (5-acetyl-2-butoxy-3-pyridyl) -3-ethyl-2- (1-ethyl-3-azetidinyl) -2, 6-dihydro-7H-pyrazolo [4, 3-d ] pyrimidin-7-one and 5- [ 2-ethoxy-5- (4-ethylpiperazine-1-yl ] pyrimidin-7-one -methylsulfonyl) pyridin-3-yl ] -3-ethyl-2- [ 2-methoxyethyl ] -2, 6-dihydro-7H-pyrazolo [4, 3-d ] pyrimidin-7-one.

Combinations of the compounds of the present invention and other therapeutic agents may be administered separately, sequentially or simultaneously.

Since, for example, for the treatment of a particular disease or condition, it may be desirable to administer a combination of active compounds, it is also within the scope of the present invention that two or more pharmaceutical compositions, at least one of which contains a compound of the present invention, may conveniently be combined in a kit suitable for the simultaneous administration of the compositions.

The kit of the invention thus comprises two or more separate pharmaceutical compositions, at least one of which contains a compound of formula (I) according to the invention, and means for separately retaining the compositions, such as a container, a separate bottle or a separate foil packet. An example of such a kit is the familiar blister pack (blister pack) used for packaging tablets, capsules, etc.

The kits of the invention are particularly suitable for administration in different dosage forms, e.g. oral and parenteral, for administration of the individual compositions at different dosage intervals, or for stepwise instillation of the individual compositions one to the other. To aid compliance, the kit typically includes instructions for administration and may be equipped with a so-called memory aid.

The compounds of the invention for pharmaceutical use may be administered as crystalline or amorphous products. They can be obtained, for example, as solid suppositories, powders or films, by methods such as precipitation, crystallization, freeze drying, spray drying or evaporative drying. Microwave or radio frequency drying may be used for this purpose. Suitable formulations of the compounds of the invention may be in hydrophilic or hydrophobic matrices, ion exchange resin complexes, coated or uncoated forms and other types as desired, as described in US 6,106,864.

The compounds of the present invention may be administered alone or in combination with one or more other drugs (or as any combination thereof). Typically they will be administered in the form of a formulation in combination with one or more suitable pharmaceutically acceptable excipients. The term "excipient" as used herein describes any component other than a compound of the present invention. The choice of excipient will depend largely on the particular mode of administration, the effect of the excipient on solubility and stability, and the nature of the dosage form. If appropriate, auxiliaries may be added. The auxiliary agent is preservative, antioxidant, flavoring agent or coloring agent.

Pharmaceutical compositions suitable for delivery of the compounds of the invention and methods suitable for their preparation will be apparent to those of ordinary skill in the art. These compositions and their preparation can be found, for example, in Remingtons Pharmaceutical Sciences, 19 th edition (Mack Publishing Company, 1995).

The compounds of the invention may be administered orally. Oral administration may involve swallowing, so the compound enters the gastrointestinal tract or may be administered orally or sublingually, whereby the compound enters the blood stream directly from the mouth.

Formulations suitable for oral administration include solid formulations such as tablets, capsules containing microparticles, liquids or powders, multiparticulates and nanoparticles, gels, films (including binders), powders, beads (ovules), elixirs, lozenges (including liquid-filled), chewables (chews), solid solutions, liposomes, suspensions, sprays and liquid formulations.

Liquid preparations include suspensions, solutions, syrups and elixirs. The formulations may be presented as a filler in soft or hard capsules and typically comprise a carrier, for example water, ethanol, polyethylene glycol, polypropylene glycol, methyl cellulose or a suitable oil, and one or more emulsifying agents and/or suspending agents. Liquid formulations can also be prepared, for example, by reconstitution from a solid in a sachet.

The compounds of the present invention may also be administered as osmotic dosage forms or in the form of high energy dispersions or as coated particles or fast dissolving, fast disintegrating dosage forms as described in Expert Opinionin Therapeutic Patents, 11(6), 981-986(2001) by Liang and Chen.

For tablet dosage forms, depending on the dosage, the drug may comprise 1 to 80 wt%, more particularly 5 to 60 wt% of the dosage form. In addition to the drug, tablets generally contain a disintegrant. Examples of disintegrants include sodium starch glycolate, sodium carboxymethylcellulose, calcium carboxymethylcellulose, croscarmellose sodium, crospovidone, polyvinylpyrrolidone, methylcellulose, microcrystalline cellulose, lower alkyl-substituted hydroxypropyl cellulose, starch, pregelatinized starch, and sodium alginate. Generally, the disintegrant will comprise from 1 wt% to 25 wt%, preferably from 5 wt% to 20 wt% of the dosage form.

Binders are commonly used to impart cohesive properties to the tablet formulation. Suitable binders include microcrystalline cellulose, gelatin, sugars, polyethylene glycol, natural and synthetic resins, polyvinyl pyrrolidone, pregelatinized starch, hydroxypropyl cellulose and hydroxypropyl methyl cellulose. Tablets may also contain diluents such as lactose (monohydrate, spray-dried monohydrate, anhydrate, etc.), mannitol, xylitol, glucose, sucrose, sorbitol, microcrystalline cellulose, starch and dibasic calcium phosphate dihydrate.

The tablets may also optionally contain surfactants such as sodium lauryl sulfate and Tween 80 and glidants such as silicon dioxide and talc. If present, the surfactant may comprise 0.2 wt% to 5 wt% of the tablet, and the glidant may comprise 0.2 wt% to 1 wt% of the tablet.

Tablets may also typically contain lubricating agents such as magnesium stearate, calcium stearate, zinc stearate, sodium stearyl fumarate, and mixtures of magnesium stearate and sodium lauryl sulfate. The lubricant will generally comprise from 0.25% to 10%, preferably from 0.5% to 3% by weight of the tablet.

Other useful components include antioxidants, coloring agents, flavoring agents, preservatives, and taste-masking agents.

Exemplary tablets contain up to about 80% drug, about 10% to about 90% binder, about 0% to about 85% diluent, about 2% to about 10% disintegrant, and about 0.25% to about 10% lubricant.

The tablet blend may be formed into tablets either directly or by roller compression. The tablet blend or blend portions may optionally be wet, dry or melt granulated, melt congealed or extruded prior to tableting. The final formulation may comprise one or more layers, and may be coated or uncoated; it may also be encapsulated.

Tablet formulations are described in "Pharmaceutical Dosage Forms: tablets, Vol.1 ", H.Lieberman and L.Lachman, Marcel Dekker, N.Y., N.Y., 1980(ISBN 0-8247-6918-X).

Solid compositions for oral administration may be formulated for immediate and/or modified release. Modified release compositions include delayed release, sustained release, pulsed release, controlled release, targeted release and programmed release. Suitable modified release compositions suitable for the purposes of the present invention are described in U.S. patent 6.106,864. Details of other suitable delivery techniques such as high energy dispersions and osmotic and coated particles are found in Verma et al Pharmaceutical Technology On-line, 25(2), 1-14 (2001).

Solid compositions of a similar type may also be employed as fillers in capsules, such as gelatin, starch or HPMC capsules. Preferred excipients in this regard include lactose, starch, cellulose, lactose or high molecular weight polyethylene glycols. The liquid compositions may be used as fillers in soft or hard capsules, for example gelatin capsules. For aqueous and oily suspensions, solutions, syrups and/or elixirs, the compounds of the invention may be combined with various sweetening or flavouring agents, colouring matter or dyes, with emulsifying and/or suspending agents, and with diluents such as water, ethanol, propylene glycol, methylcellulose, alginic acid or sodium alginate, glycerol, hydrocolloid agents and combinations thereof. In addition, formulations containing these compounds and excipients prior to use may be presented as a dry product for formulation with water or other suitable carrier.

The compounds of the invention may also be administered by injection, i.e., intravenous, intramuscular, intradermal, or intraperitoneal, intraarterial, intrathecal, intraventricular, intraurethral, intrasternal, intracranial, intraspinal, or subcutaneous injection, or they may be administered by infusion, needle (including microneedle) syringe, needleless syringe, or by an implanted injection technique. For these parenteral administrations, they are usually used in the form of a sterile aqueous solution, suspension or emulsion (or system, so that micelles may be included), which may contain other substances known in the art, for example, sufficient salts or carbohydrates, such as glucose, to render the solution isotonic with blood. The aqueous solution should be suitably buffered if necessary (preferably to a pH of 3 to 9). For some forms of parenteral administration, they may be used in the form of a sterile non-aqueous system, for example a fixed oil, including mono-and diglycerides and fatty acids (including OLEIC acids). Preparation of suitable parenteral formulations under sterile conditions, e.g., freeze-drying, is readily carried out by standard pharmaceutical techniques well known to those of ordinary skill in the art. Alternatively, the active ingredient may be in powder form for constitution with a suitable vehicle, e.g., sterile, pyrogen-free water, before use.

The solubility of the compounds of formula (I) used in the preparation of parenteral solutions may be enhanced by the use of appropriate formulation techniques, for example the introduction of solubility enhancers.

Compositions for parenteral administration may be formulated for immediate and/or modified release. The compounds of the invention can therefore be formulated in more solid form for administration as an embedded sustained release agent (immunophenodepot) to provide long-term release of the active compound.

The compounds of the invention may also be administered intranasally or by inhalation. They are conveniently delivered in dry powder form (either alone, as a mixture, e.g. with lactose, or mixed component particles, e.g. a dry blend with phospholipids) from a dry powder inhaler or an aerosol nebulizer by means of a pressurised container, pump, eductor, nebulizer (preferably using an electro-hydrodynamic nebulizer to produce a fine mist) or nebulizer (nebuliser), with or without the use of a suitable propellant, e.g. dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, a hydrofluoroalkane, such as 1, 1, 1, 2-tetrafluoroethane (HFA 134A [ trade mark ]) or 1, 1, 2, 3, 3, 3-heptafluoropropane (HFA 227EA [ trade mark ]), carbon dioxide, another perfluorocarbon such as Perflubron (trade mark) or other suitable gas. For intranasal use, the powder may include a bioadhesive agent such as chitosan or cyclodextrin.

In the case of a pressurized aerosol, the dosage unit may be determined by providing a valve to deliver a metered amount. The pressurized container, pump, sprayer or sparger contains a solution or suspension of the compound of the invention, including, for example, ethanol (aqueous ethanol) or a suitable agent for dispersion, solubilization or extended release and a propellant as a solvent, which may additionally contain a lubricant, such as sorbitol trioleate or lactic acid oligomers.

Prior to use in a dry powder formulation or suspension formulation for inhalation, the compounds of the invention will be micronized to a size suitable for delivery by inhalation (generally considered to be less than 5 microns). Micronization can be carried out by methods such as spiral jet milling, fluidized bed jet milling, processing using supercritical fluids to form nanoparticles, high pressure homogenization, or by spray drying.

Capsules (made of e.g. gelatin or HPMC), blisters and cartridges for use in an inhaler or insufflator may be formulated containing a powder mix of: the compounds of the invention, a suitable powder base such as lactose or starch and a property modifier such as 1-leucine, mannitol or magnesium stearate. Lactose can be anhydrous or in the form of a monohydrate, the latter being preferred. Other suitable excipients include dextran, glucose, maltose, sorbitol, xylitol, fructose, sucrose and trehalose.

Suitable solution formulations for use in preparing fine sprays using an electro-hydraulic-dynamic nebulizer may contain from 1 microgram to 20 milligrams of the compound of the invention per spray (per actuation) and may vary in volume from 1 microliter to 100 microliters. A typical formulation may comprise a compound of the invention, propylene glycol, sterile water, ethanol and sodium chloride. Other solvents that may be substituted for propylene glycol include glycerol or polyethylene glycol.

Suitable flavouring agents, such as menthol and levomenthol or sweeteners, such as saccharin or sodium saccharin may be added to those formulations of the invention for inhalation/intranasal administration.

Compositions for inhalation/intranasal administration may be formulated in immediate and/or modified release form using, for example, (DL) -lactic-glycolic acid (PGLA) copolymers. Modified release compositions include delayed, sustained, pulsed, controlled, targeted, and programmed release.

Alternatively, the compounds of the present invention may be applied topically, i.e., transdermally or transdermally, to the skin or mucosa. Typical formulations for this purpose include gels, hydrogels, lotions, solutions, creams, ointments, dusting powders, dressings, foams, membranous skin patches, films, implants, sponges, fibers, bandages and microemulsions. Liposomes may also be used. For these applications, the compounds of the invention may be suspended or dissolved in a mixture, for example, having one or more of the following: mineral oil, liquid petrolatum, white petrolatum, propylene glycol, polyethylene oxide polypropylene compounds, emulsifying waxes, non-volatile oils including synthetic mono-or diglycerides and fatty acids including oleic acid, water, sorbitan monostearate, polyethylene glycol, liquid paraffin, Tween 60, cetyl esters wax, crystal wax alcohol, 2-octyldodecanol, benzyl alcohol, alcohols such as ethanol may be incorporated-see for example J.Pharm.Sci., 88(10), 955-.

Other means of topical administration include delivery by: iontophoresis, electroporation, sonophoresis (sonophoresis) and needle-or microneedle injection (e.g. Powderject)^TM，Bioject^TMEtc.).

Compositions for topical administration may be formulated for immediate and/or modified release. Modified release compositions include delayed, sustained, pulsed, controlled, targeted, and programmed release.

Alternatively, the compounds of the invention may be administered rectally, for example in the form of suppositories, pessaries or enemas. They may also be administered by the vaginal route. For example, these compositions may be prepared by mixing the drug with a suitable non-irritating excipient, such as cocoa butter, synthetic glyceride esters or polyethylene glycols, which are solid at ordinary temperatures but emulsify and/or dissolve in the cavity to release the drug.

Compositions for rectal/vaginal administration may be formulated for immediate and/or modified release. Modified release compositions include delayed, sustained, pulsed, controlled, targeted, and programmed release.

The compounds of the invention may also be administered directly to the eye or ear, usually in the form of droplets of a micronized suspension or solution in isotonic, pH-adjusted sterile saline. Other formulations suitable for ocular or otic administration include ointments, biodegradable (e.g., absorbable gel sponges, collagen) or non-biodegradable (e.g., silicone) implants, films, lenses, and microparticles or vesicular systems such as niosomes or liposomes. A polymer. Such as cross-linked polyacrylic acid, polyvinyl alcohol, hyaluronic acid, cellulosic polymers (e.g., hydroxypropyl methylcellulose, hydroxyethyl cellulose, methyl cellulose), or heteropolysaccharide polymers (e.g., agarose gel) may be incorporated together as preservatives, such as benzalkonium chloride. These formulations may also be delivered using iontophoresis.

Compositions for ocular/otic administration may be formulated for immediate and/or modified release. Modified release compositions include delayed, sustained, pulsed, controlled, targeted, and programmed release.

The compounds of the present invention may also be combined with soluble macromolecular substances, such as cyclodextrins and suitable derivatives thereof or polyethylene glycol-containing polymers, in order to improve their solubility, dissolution rate, taste masking, bioavailability and/or stability for use in any of the aforementioned modes of administration.

For example, drug-cyclodextrin complexes are commonly used in most dosage forms and routes of administration. The form containing the complex may be used, and the form containing no complex may be used. As an alternative to direct complexation with the drug, the cyclodextrin may be used as an auxiliary aid, i.e. as a carrier, diluent or solubiliser. alphA-, betA-and gammA-cyclodextrins are mostly used in general, examples of which are described in WO-A-91/11172, WO-A-94/02518 and WO-A-98/55148.

The term "administering" includes delivery by viral or non-viral techniques. Viral delivery mechanisms include, but are not limited to, adenoviral vectors, adeno-associated viral (AAV) vectors, herpes viral vectors, retroviral vectors, lentiviral vectors, and baculovirus vectors. Non-viral delivery mechanisms include lipid-mediated transition infections, lipomes, immunoliposomes, lipofectins, Cationic Facial Amphiphiles (CFAs), and combinations thereof. Routes for these delivery mechanisms include, but are not limited to, mucosal, nasal, oral, parenteral, gastrointestinal, topical, or sublingual routes.

Thus, the present invention provides a pharmaceutical composition comprising a compound of formula (I) or a pharmaceutically acceptable salt, solvate, polymorph or prodrug thereof, in association with a pharmaceutically acceptable excipient, diluent or carrier.

The units of the pharmaceutical formulation are preferably in unit dosage form. In such form, the preparation is divided into unit doses containing appropriate quantities of the active ingredient. The unit dosage form can be a packaged preparation, the package containing discrete quantities of preparation, such as packeted tablets, capsules, and powders in bottles or ampoules. Moreover, the unit dosage form can be a capsule, tablet, cachet, or lozenge itself, or it can be the appropriate number of such dosage forms in the form of a package. The amount of active ingredient in a unit dose formulation can vary or be adjusted from 0.1mg to 1g depending on the particular formulation and potency of the active ingredient. In pharmaceutical applications, the drug may be administered 1 to 3 times per day, for example as 100 or 300mg capsules. In therapeutic use, the compounds used in the pharmaceutical methods of the invention are about 0.01mg to about 100mg per kg per day in the initial dose. A preferred daily dosage range is from about 0.01mg to about 100 mg/kg.

The dose is based on an average human patient weighing about 65kg to 70 kg. A physician will be able to easily determine dosages for patients with weights outside this range, such as infants and elderly people. However, the dosage may vary based on the needs of the patient, the severity of the condition being treated and the compound used. Determination of the appropriate dosage for a particular condition is within the skill of one of ordinary skill in the art. Typically, treatment is initiated at a smaller dose than the optimal dose of the compound. Thereafter, the dosage is increased by a small increase until the optimum effect under the condition is reached. For convenience, the total daily dose may be divided into several administrations throughout the day if desired.

The pharmaceutical compositions of the present invention may, if desired, contain one or more compatible therapeutic agents. In particular, the composition may be combined with any one or more compounds useful for treating pain (such as those listed above). Accordingly, the present invention provides pharmaceutical compositions comprising a compound of formula (I), one or more other pharmacologically active agents and one or more pharmaceutically acceptable carriers.

For the avoidance of doubt, the term "treatment" herein includes curative, palliative and prophylactic treatment.

General procedure

Compounds of formula (I) can be synthesized using various methods given below:

according to the first process, (A), the compound of formula (I) can be prepared by deprotection of the compound of formula (II), (III) or (IV) by a conventional method, for example, acidolysis with a strong acid such as trifluoroacetic acid or hydrochloric acid in a suitable solvent such as dioxane or dichloromethane

Wherein R, X and Y are as described above for formula (I) and R1 is a suitable carboxylic acid protecting group, e.g. C_1-16Alkyl, and PG is a suitable protecting group such as tert-butoxycarbonyl.

The compounds of formula (I) may be prepared directly from the compounds of formula (III) by hydrolytic cleavage.

The compound of formula (II) may be prepared by hydrolysis of the ester functionality of compound (III),

wherein X, Y, R, PG and R₁As defined above, and the hydrolysis is promoted by an alkali metal hydroxide, such as lithium hydroxide, in a suitable solvent, such as aqueous dioxane.

The compounds of formula (III) may be prepared by the following process:

i) the compound of formula (VI) is reacted with a compound RYX-H in a suitable solvent, such as DMF, at a temperature of 20 to 140 ℃ using a suitable base, such as an alkali metal salt, such as K₂CO₃Or an alkali metal hydride, such as NaH,

wherein Z is a suitable leaving group such as mesylate, tosylate, triflate or halogen.

ii) when RYX-is ArO-, wherein Ar is an optionally substituted aryl or heteroaryl ring, the compound of formula (VII) is reacted with a compound of formula ArOH in a suitable solvent, e.g. THF, at a temperature of 25-60 ℃ using Mitsunobu conditions of the appropriate azidodicarboxylate, e.g. DIAD and triphenylphosphine or tributylphosphine.

iii) hydrogenolysis of the compound of formula (VIII) with a suitable catalyst such as palladium on carbon.

The compounds of formula (VIII) may be prepared from compounds of formula (XII), for example in NBu, by addition of an organometallic compound in the presence of a suitable catalyst and additive in a suitable solvent, for example 1: 1 THF: 1-methyl-2-pyrrolidone₄l, palladium catalyst and phosphine ligand, and adding benzyl zinc bromide to prepare the catalyst.

The compounds of formula (XII) are prepared from the compounds of formula (X) by addition of the appropriate base at-78 ℃ to-20 ℃ followed by the addition of a triflating agent, for example n-butyllithium in a suitable solvent such as THF followed by the addition of triflic anhydride.

iv) when X is CH₂When desired, the compound of formula (IX) is hydrolysed with a suitable catalyst, for example palladium on carbon.

The compound of formula (IX) can be prepared from the compound of formula (X) using a Wittig reaction in which a ylide is formed at room temperature from a suitable phosphonium salt and a base such as 1M tBuOK/THF or sodium tert-amylate in toluene or dichloromethane.

Under basic conditions, e.g. THF/H₂The aqueous lithium hydroxide solution in O hydrolyzes the compound of formula (IX) to the compound of formula (XVII). At room temperature, using standard coupling agents such as DCC, DMAP and an appropriate alcohol such as menthol (R) in methylene chloride₂) The compound of formula (XVIII) is prepared from the compound of (XVII).

At room temperature, under a hydrogen atmosphere of 15psi, using a suitable catalyst such as PtO in EtOAc and/or toluene₂And, hydrogenating the compound of formula (XVIII) for 1 to 18 hours to obtain the compound of formula (XIV).

Deprotecting the compound of formula (XIV) completely, suitably using 6M hydrochloric acid, at 60 ℃ to 120 ℃ for 18 hours according to method A, to give the compound of formula (I) wherein X is CH₂。

Alternatively, the compounds of formula (VIII) and (IX) may be prepared by acid-catalyzed dehydration of the compound of formula (XI).

The compounds of formula (XI) may be prepared by adding the organometallic to the compound of formula (VIII), for example benzylmagnesium bromide, in a suitable solvent such as THF at a temperature of-78 deg.C to 20 deg.C.

v) when Y is O and X is CH₂When the compound of formula (XVI) is reacted with a compound of formula R-OH, Mitsunobu conditions are used.

The compound of formula (XVI) can be prepared by hydroboration of the compound of formula (XV).

The compounds of formula (XV) are prepared from compounds of formula (XIII) by hydrolysis of the ester functionality to give compounds of formula (XIV) followed by re-esterification.

The compound of formula (XIII) can be prepared from the compound of formula (X) using an appropriate methylene Wittig reagent, such as methyltriphenylphosphonium bromide, and a base, such as potassium tert-butoxide, in an appropriate solvent, such as toluene.

It is also possible to use hydroboration agents such as BH₃The compound of formula (XVI) is prepared by reduction of a carboxylic acid of formula (XVII) in a suitable solvent such as THF at a temperature of 0-30 ℃.

Can be prepared by using appropriate conditions, such as ruthenium chloride, and sodium periodate, in a solvent mixture, such as H2O, EtOAc and CH₃CN in room temperature, aryl oxidation of the compound of formula (XVIII) produces the compound of formula (XVII).

For the above general methods, one of ordinary skill in the art will readily appreciate that when protecting groups are present, these protecting groups are generally exchangeable with other protecting groups having similar properties, e.g., when an amine is described as being protected with a tert-butoxycarbonyl group, it may be exchanged with any suitable amine protecting group.

The invention is illustrated by the following non-limiting examples and intermediates, in which the following abbreviations are used:

THF tetrahydrofuran

DMF dimethyl formamide

DIAD Azobicarboxylic acid diisopropyl ester

EtOAc ethyl acetate

DCM dichloromethane

rt Room temperature

MeOH methanol

ETOH ethanol

TFA trifluoroacetic acid

BOC tert-butoxycarbonyl

Example 1

(2S, 4S) -4- (benzylsulfanyl) -pyrrolidine-2-carboxylic acid

To a solution of di-tert-butyl (2S, 4S) -4-benzylthio-pyrrolidine-1, 2-dicarboxylate (preparation 2, 130mg, 3.3mmol) in dichloromethane (2.5ml) was added trifluoroacetic acid (2.5ml), and the mixture was stirred at room temperature under a nitrogen atmosphere for 36 hours. The solvent was removed under reduced pressure and the residue was purified by ion exchange chromatography using Dowex^TMThe 50WX8-200 resin, first eluted with water and then 10% ammonia gave the title compound (66mg, 75%) as a white solid.

¹H-NMR(400MHz，D₂O)δ＝1.88-1.98(1H，m)；2.45-2.56(1H，m)；3.07-3.13(1H，m)；3.22-3.38(2H，m)；3.66-3.74(2H，s)；3.93-4.01(1H，m)；7.11-7.29(5H，m)

LRMS (electrospray): m/z [ MH⁺]238；[MNa+]260；[MH^-]236

Microanalysis: c, 59.36; h, 6.33; n, 5.77.C₁₂H₁₅NO₂S.0.3H₂O requires C, 59.38; h, 6.48; n, 5.77

Example 2

(2S, 4S) -4- [ (4-chlorobenzyl) oxy]-pyrrolidine-2-carboxylic acid

Reacting (2S, 4S) -1- (tert-butoxycarbonyl)4- [ (4-chlorobenzyl) oxy) phenyl]-2-Pyrrolidinecarboxylic acid (preparation 4, 96mg, 0.38mmol) was dissolved in dichloromethane (5 ml). Trifluoroacetic acid (5ml) was added to the solution, and the mixture was allowed to stand overnight at room temperature. The reaction mixture was partitioned between dichloromethane (25ml) and water (25 ml). The aqueous layer was separated, washed with dichloromethane (25ml) and evaporated to dryness. Dowex was used as the product^TMPurification on 50WX8-200 resin eluting first with water and then with 9: 1 water: ammonia gave the title compound (5mg, 5% yield) as a white solid.

¹H-NMR(400MHz，CD₃OD)δ＝2.4-2.5(m，1H)，2.6-2.7(m，1H)，3.4-3.5(m，1H)，3.6-3.7(m，1H)，4.5-4.7(m，4H)，7.3-7.5(m，4H).

LCMS (electrospray): m/z [ M ]^-]254

Example 3

(2S, 4S) -4- [ (4-bromophenylthio-pyrrolidine-2-carboxylic acid)

(2S, 4S) -4- (4-bromo-phenylthio)) -pyrrolidine-1, 2-dicarboxylic acid 1-tert-butyl ester (preparation 7, 54mg, 0.14mmol) was dissolved in 4M HCl in dioxane and stirred at room temperature for 2 h. The solvent was removed in vacuo to give a milky solid (32mg, 76%).

¹H-NMR(400MHz，CD₃OD)δ＝2.20(1H，m)，2.83(1H，m)，3.32(1H，m)，3.70(1H，m)，4.15(1H，m)，4.50(1H，m)，7.40(2H，d)，7.55(2H，m).

LRMS (electrospray): m/z [ MH⁺]302，304.

Microanalysis: c, 39.01; h, 4.23; n, 4.14.C₁₁H₁₂NO₂SBr.0.9HCl requires C, 39.44; h, 3.88; n, 4.18.

Example 4

(2S, 4S) -4-Phenylthio-pyrrolidine-2-carboxylic acid

Starting from the title compound of preparation 8, the title compound was prepared by the method of example 3. Yield 60% and title compound as white solid.

¹H-NMR(400MHz，CD₃OD)δ＝2.19(1H，m)，2.80(1H，m)，3.34(1H，m)，3.70(1H，m)，4.10(1H，m)，4.56(1H，m)，7.030-7.60(5H，m).

LCMS (electrospray): m/z [ MH⁺]224.

Microanalysis: c, 48.95; h, 5.50; n, 4.97.C₁₁H₁₃NO₂S.HCl.0.5H₂O requires C, 49.16; h, 5.63; n, 5.21.

Example 5

(2S, 4S) -4- [ 2-fluorophenoxy ] -pyrrolidine-2-carboxylic acid

From the title compound of preparation 10, the title compound was prepared by the method of example 3 in 74% yield.

¹H-NMR(400MHz，MeOD)：δ＝2.60-2.76(m，2H)，3.57-3.65(m，1H)，3.75(d，2H)，4.56-4.64(m，1H)，4.85(s，3H)，5.18-5.24(m，1H)，6.98-7.19(m，4H).

LRMS (electrospray): [ M-1]]224，[MH⁺]226.

Microanalysis: the following are found: c, 50.38; h, 4.95; n, 5.29% C₁₁H₁₂FNO₃C, 50.49; h, 5.01, N, 5.35%

Example 6

(2S, 4S) -4- [ (4-chlorophenoxy ] -pyrrolidine-2-carboxylic acid

The BOC protected product from preparation 12 (250mg, 0.73mmol) was stirred for 2 hours at 0 deg.C in 4M HCl in dioxane (5 ml). Diethyl ether (10ml) was added, the resulting precipitate was filtered off, and washed with diethyl ether to give the title compound (178mg, 87%).

¹H-NMR(400MHz，MeOD)：δ＝2.59-2.71(m，2H)，3.56-3.72(m，2H)，4.57-4.66(m，1H)，4.82-4.93(M，3H)，5.17-5.25(m，1H)，6.88-6.98(m，2H)，7.26-7.36(m，2H).

LRMS (electrospray): [ M-1]]240，[MH⁺]242，[MNa⁺]264.

Microanalysis: c, 47.48; h, 4.71; n, 4.92.C₁₁H₁₂ClNO₃HCl requires C, 47.50; h, 4.71; n, 5.04%

Example 7

(2S, 4S) -4- [ 2-isoquinolinyloxy]-pyrrolidine-2-carboxylic acid

(2S, 4S) -4- (isoquinoline-7-yloxy) -pyrrolidine-1, 2-dicarboxylic acid 1-tert-butyl ester (preparation 13, 120mg, 0.29mmol) was stirred at room temperature in TFA (3ml) for 4.5 h. The solvent was removed in vacuo and triturated with diethyl ether to give a very hygroscopic solid which was redissolved in 2N HCl (3ml) and stirred at room temperature for 1 hour. The solution was washed 1 time with diethyl ether (5ml) and the water was evaporated to give a foam. Trituration with ether gave the title compound as a glass (24mg, 28%).

¹H-NMR(400MHz，CH₃OD)：δ2.68-2.80(m, 1H), 2.82-2.97(m, 1H), 3.75-3.91(m, 2H), 4.62-4.75(m, 1H), 4.75-4.96(m, 5H switchable), 5.48-5.60(m, 1H), 7.75-7.81(m, 1H), 7.98-8.02(m, 1H), 8.26(d, 1H), 8.39-8.55(m, 2H), 9.64(s, 1H)

LRMS (electrospray) [ M-1]]257，[MH⁺]259

Example 8

(2S, 4S) -4- (3-chloro-phenoxy) -pyrrolidine-2-carboxylic acid

From the title compound of preparation 15, prepared by the method of example 6 and washed with diethyl ether (2 × 20ml) to give a white solid (52mg, 93%).

¹H NMR(400MHz，CD₃OD)：δ＝2.65(m，2H)，3.60(dd，1H)，3.70(d，1H)，4.60(dd，1H)，5.02(m，1H)，6.88(m，1H)，6.97(s，1H)，7.03(d，1H)，7.29(dd，1H).

LRMS (electrospray [ MH ]⁺]242，[M-1]240.

Microanalysis: c, 46.97; h, 4.70; n, 4.90.C₁₁H₁₂ClNO₃.HCl.0.1H₂O requires C, 47.20; h, 4.75; and N, 5.00.

Example 9

(2S, 4S) -4- (benzyloxy) -pyrrolidine-2-carboxylic acid

(2S, 4S) -1- (tert-butoxycarbonyl) -4- (benzyloxy) -pyrrolidine-2-carboxylic acid (preparation 17, 150mg, 0.47mmol) was dissolved in dichloromethane (5 ml). Trifluoroacetic acid (5ml) was added to the solution, and the mixture was stirred at room temperature overnight. The reaction mixture was partitioned between dichloromethane (25ml) and water (25 ml). The aqueous layer was removed, washed with dichloromethane (25ml) and evaporated to a dry substance. The product was purified using an ion exchange column (Dowex50WX8-200 resin) eluting first with water and then with 9: 1 water: ammonia to give the title compound (34mg, 33% yield) as a white solid.

¹H-NMR(400MHz，CD₃OD)δ＝2.3-2.5(m，1H)，3.1-3.18(m，1H)，3.4-3.5(d，1H)，3.9-3.95(m，1H)，4.2(s，1H)，4.4-4.55(dd，3H)，7.2-7.4(m，5H).

LCMS (electrospray): m/z [ MNa ]⁺]244.

Example 10

(2S, 4S) -4- (3-fluoro-benzyl) -pyrrolidine-2-carboxylic acid monohydrochloride

4- (3-fluoro-benzyl) -pyrrolidine-1, 2-dicarboxylic acid 1-tert-butyl ester 2- (2-isopropyl-5-methyl-cyclohexyl) ester (preparation 35, 0.91g, 1.96mmol) was dissolved in toluene (2 ml). Adding 6N saltAcid (50ml) and stirred under reflux for 18 h. The reaction mixture was cooled to room temperature and extracted with ethyl acetate (3X 20 ml). The aqueous layer was concentrated by evaporation under reduced pressure to give the title compound (417mg, 81%) as a white solid.¹H-NMR showed the cis: trans diastereomer in a 7: 1 ratio and the product was recrystallized from isopropyl alcohol to give the title compound (170mg, 65%) which was determined by NMR to have a cis: trans ratio of 14: 1.

¹H-NMR(400MHz，CD₃OD): (diastereomer 2S, 4S: 2S, 4R (14: 1)): δ is 1.85(q, 1H), 2.51(quin, 1H), 2.69-2.85(m, 3H), 3.07(t, 1H), 3.41(dd, 1H), 4.38 and 4.48(t, 1H), 6.90-7.04(m, 3H), 7.32(q, 1H).

LRMS(APCI)：m/z[MH]⁺224.

[α]_D ²⁵-1.27 ° (c ═ 9.00 in methanol).

Microanalysis: c, 55.56; h, 5.81; n, 5.34%. C₁₂H₁₄FNO₂HCl requires C, 55.50; h, 5.82; and N, 5.39 percent.

Example 11

(2S, 4S) -4- (2, 3-difluoro-benzyl) -pyrrolidine-2-carboxylic acid monohydrochloride

Starting from the title compound of preparation 37, the title compound was prepared by the method of example 10 and purified by recrystallization from acetone/diethyl ether to give a white solid of the title compound as a mixture of diastereomers (2S, 4S: 2S, 4R (12: 1))¹H-NMR (500mg, 60%) measurement.

¹H-NMR(400MHz，CD₃OD) (diastereomer cis: trans (12: 1)): delta 0.80-1.90(m，0.92H)，2.12-2.20(m，0.08H)，2.28-2.36(m，0.08H)，2.49-2.58(q，0.92H)，2.66-2.81(m，1H)，2.83-2.95(m，2H)，3.02-3.13(t，1H)，3.46(dd，1H)，4.40(dd，0.92H)，4.48-4.54(m，0.08H)，7.03-7.20(m，3H).

LRMS (electrospray): m/z [ M + H]⁺242.

Microanalysis: c, 51.42; h, 5.08; n, 5.01%. C₁₂H₁₃NO₂F₂HCl requires C, 51.90; h.5.08; and N, 5.04%.

Example 12

(2S, 4S) -4- (2, 5-difluoro-benzyl) -pyrrolidine-2-carboxylic acid monohydrochloride

The title compound was prepared by the method of example 10 starting from the title compound of preparation 36.

¹H-NMR(400MHz，CD₃OD): (diastereomer 2S, 4S: 2S, 4R (26: 1)): δ — 1.86(q, 1H), 2.51-2.54(m, 1H), 2.75-2.83(m, 3H), 3.09(t, 1H), 3.45(q, 1H), 4.39 and 4.49(2t, 1H) 26: 1, 7.00-7.14(m, 3H).

LRMS(APCI)：m/z[MH]⁺242.

Microanalysis: c, 50.18; h, 4.94; n, 4.83%. C₁₂H₁₃F₂NO₂HCl requires C, 51.90; h, 5.08; and N, 5.04%.

[α]_D ²⁵-0.22 ° (c ═ 1.84 in methanol).

Example 13

(2S, 4S) -4-cyclohexylmethyl-pyrrolidine-2-carboxylic acid monohydrochloride

4-Cyclohexylmethyl-pyrrolidine-1, 2-dicarboxylic acid 1-tert-butyl ester 2- (2-isopropyl-5-methyl-cyclohexyl) ester (preparation 38, 316mg, 0.70mmol) was dissolved in toluene (2 ml). 6N hydrochloric acid (50ml) was added and stirred under reflux for 72 hours. The reaction mixture was cooled to room temperature and extracted with ethyl acetate (3X 20 ml). Concentration of the aqueous layer by evaporation under reduced pressure gave the title compound as a white solid (80mg, 48%).

¹H-NMR(400MHz，CD₃OD): (diastereomer 2S, 4S: 2S, 4R (6: 1)): δ is 0.83-1.00(m, 2H), 1.13-1.40(m, 6H), 1.62-1.81(m, 6H), 2.48(m, 2H), 2.90(t, 1H), 3.48(t, 1H), 4.32 and 4.42(2t, 1H).

LRMS(APCI)：m/z[MH]⁺ 212.

[α]_D ²⁵-1.86 ° (c ═ 2.04 in methanol)

Example 14

(2S, 4S) -4- (3-methoxy-benzyl) -pyrrolidine-2-carboxylic acid monohydrochloride

Starting from the title compound of preparation 39, the title product was prepared by the method of example 10.

¹H-NMR(400MHz，CD₃OD): (diastereomer 2S, 4S: 2S, 4R (15: 1)): δ is 1.79-1.89(m, 1H), 2.47-2.52(m, 1H), 2.68-2.77(m, 3H), 3.06(t, 1H), 3.36(t, 1H), 3.39(s, 3H), 4.37 and 4.47(t, 1H))，6.81(d，3H)，7.22(t，1H).

LRMS(APCI)：m/z[MH]⁺236.

Microanalysis: c, 56.77; h, 6.62; n, 5.06%. C₁₃H₁₇NO₃HCl requires C, 57.46; h, 6.68; and N, 5.15 percent.

[α]_D ²⁵-6.90°(c＝3.1，MeOH).

Example 14A

(2S, 4S) -4- (3-methoxy-benzyl) -pyrrolidine-2-carboxylic acid monohydrochloride can also be prepared by the method in J.Ezquerra, C.Pedegrel, B.YRurtagoyena and A.Rubio in J.Org.Chem.1995, 60, 2925-.

Example 15

(2S, 4S) -4- (3-fluoro-phenoxymethyl) -pyrrolidine-2-carboxylic acid

Di-tert-butyl 4- (3-fluoro-phenoxymethyl) -pyrroline-1, 2-dicarboxylate (preparation 44, 475mg, 1.2mmol) was dissolved in a solution of anhydrous hydrogen chloride in dioxane (4M, 15ml) and stirred at 50 ℃ under a nitrogen atmosphere for 1 hour. The solvent was removed under reduced pressure and the resulting semi-solid triturated with ethyl acetate to give a white solid which was recrystallized from ethyl acetate/isopropanol as a mixture of diastereomers (. about.5: 12S, 4S: 2S, 4R) to give the title compound as the hydrochloride salt as a white solid (90mg, 35%)

¹H-NMR(400MHz，CD₃OD)：δ＝2.04-2.09(m，0.8H)；2.33-2.47(m，0.4H)；2.65-2.75(m，0.8H)；2.88-3.00(m，1H)；3.33-3.40(m，1H)；3.52-3.60(m，0.8H)；3.60-3.68(0.2H)；3.96-4.04(m，1H)；4.04-4.12(m，1H)；4.42-4.51(m，0.8H)；4.40-4.56(m，0.2H)；6.65-6.80(m，3H)，7.21-7.30(m，1H)

LRMS (electrospray): [ M +1] 240; [ M +23] 262; [ M-1]238.

The following compounds can be obtained by a method similar to example 15:

example 16

(2S, 4S) -4- (2, 5-difluoro-phenoxymethyl) -pyrrolidine-2-carboxylic acid;

example 17

(2S, 4S) -4- (2, 3-difluoro-phenoxymethyl) -pyrrolidine-2-carboxylic acid;

example 18

(2S, 4S) -4- (3-methoxy-phenoxymethyl) -pyrroline-2-carboxylic acid

Example 19

(2S, 4S) -4- (3-chloro-phenoxymethyl) -pyrrolidine-2-carboxylic acid

(2S, 4S) -4- (3-chloro-phenoxymethyl) -pyrrolidine-1, 2-dicarboxylic acid di-tert-butyl ester (preparation 46,67mg, 0.16mmol) was dissolved in a solution of anhydrous hydrogen chloride in dioxane (4M, 5ml) and stirred at room temperature for 18 h. The solvent was removed under reduced pressure and the residue triturated with ethyl acetate to give the title compound as the hydrochloride salt as a white solid (13mg, 27%)

¹H-NMR(400MHz，CD₃OD)：δ＝2.07-2.18(m，1H)；2.63-2.74(m，1H)；2.88-3.00(m，1H)；3.32-3.40(m，1H)；3.52-3.61(m，1H)；3.96-4.04(m，1H)；4.04-4.10(m，1H)；4.42-4.51(t，1H)；6.82-6.89(d，1H)；6.80-7.00(m，2H)；7.20-7.28(t，1H)

LRMS (electrospray): [ M +1] 256; [ M +23] 278; [ M-1]254

Example 20

(2S, 4S) -4(2, 3-dihydro-benzofuran-6-yloxy) -pyrrolidine-2-carboxylic acid

The title compound was prepared by the method of example 3 as a pale yellow solid in 100% yield.

¹H-NMR(400MHz，D₂O)：δ＝2.35-2.56(m，2H)；2.86-3.04(m，2H)；3.35-3.65(m，2H)；4.10-4.26(m，3H)；4.97-5.05(m，1H)；6.20-6.36(m，2H)；7.02(d，1H).

LRMS (electrospray): [ MH⁺]250

Microanalysis: the following are found: c54, 16; h, 5.78; n, 4.72%. C₁₃H₁₅NO₄.HCl.0.15H₂O requires C, 54.14; h, 5.70; and N, 4.86.

Example 21

(2S, 4S) -4-3-chloro-phenylamino) -pyrrolidine-2-carboxylic acid

4- (3-chloro-phenylamino) -pyrrolidine-1, 2-dicarboxylic acid 1-tert-butyl ester (preparation 41, 155mg, 0.456MMOL) was stirred in 4M HCl in dioxane (4ml) at 0 ℃ for 2 h. Ether (4ml) was added and the resulting white hygroscopic solid was filtered off and dried under vacuum at 40 ℃ to give the title compound (90mg, 60.3%).

¹H-NMR(400MHz，CD₃OD)：2.20-2.29(m，1H)；2.95-3.05(m，1H)；3.28-3.39(m，2H)；4.22-4.31(m，1H)；4.45-4.55(m，1H)；4.90(s，5H)；6.62(d，1H)；6.70-6.75(m，2H)；7.13(t，1H).

LRMS (electrospray): [ M-1]239.

Microanalysis: the following are found: c, 40.37; h, 5.07; n, 8.46%. C₁₁H₁₃ClN₂O₂.2HCl.0.75H₂O requires C, 40.39; h, 5.08; n, 8.56.

Preparation 1

(2S, 4R) -4- (toluene-4-sulfonyloxy) -pyrrolidine-1, 2-dicarboxylic acid di-tert-butyl ester

To 20ml CH₂Cl₂To a solution of di-tert-butyl (2S, 4R) -4-hydroxy-pyrrolidine-1, 2-dicarboxylate (CAS Reg.No.170850-75-6) (1g, 3.48mmol) were added pyridine (3.9ml) and p-toluenesulfonyl chloride (0.7g, 3.67mmol), and the mixture was stirred at room temperature under a nitrogen atmosphere for 72 hours. In thatThe solvent was removed under reduced pressure and the residue was dissolved in EtOAc (100ml) and washed with saturated citric acid solution (50ml) and then water (50 ml). The organic phase was dried (magnesium sulfate), filtered and evaporated under reduced pressure. The residue was purified by column chromatography, eluting with ethyl acetate: heptane (3: 10), to give the title compound (1.5g, 98%) as a colorless gum.

¹H-NMR(400MHz，CDCl₃)δ＝1.39-1.49(18H，m)，2.01-2.16(1H，m)，2.33-2.6(4H，m)，3.50-3.64(2H，m)，4.20-4.29(1H，m)，4.96-5.06(1H，m)；7.31-7.40(2H，m)，7.65-7.80(2H，m).

LRMS (electrospray): m/z [ MH⁺]464，[MH^-]440

Preparation 2

(2S, 4S) -4-benzylthio-pyrrolidine-1, 2-dicarboxylic acid di-tert-butyl ester

To a solution of preparation 1(200mg, 4.53mmol) in ethanol (10ml) were added benzyl mercaptan (0.107ml, 8.86mmol) and potassium tert-butoxide (101mg, 8.86mmol) under a nitrogen atmosphere, and the mixture was stirred at room temperature for 18 hours. The solvent was removed under reduced pressure and the residue was dissolved in EtOAc (25mL) and washed with water (10 mL). The organic phase was dried (magnesium sulfate), filtered and evaporated under reduced pressure. The residue was purified by column chromatography, eluting with heptane: ethyl acetate (9: 1) to give the title compound (130mg, 73%) as a colorless oil.

¹H-NMR(400MHz，CDCl₃)δ＝1.38-1.50(18H，m)，1.80-1.90(1H，m)，2.44-2.55(1H，m)，3.00-3.29(2H，m)，3.70-3.78(2H，s)，3.84-3.95(1H，m)，4.04-4.16(1H，m)，7.27-7.34(5H，m).

LRMS (electrospray): m/z [ MNa ]⁺]416

Preparation 3

(2S, 4S) -4- (4-chloro-benzyloxy) -pyrrolidine-1, 2-dicarboxylic acid 1-tert-butyl ester 2-methyl ester

(2S, 4S) -4-hydroxy-pyrrolidine-1, 2-dicarboxylic acid 1-tert-butyl ester 2-methyl ester (CAS Reg. No.227935-38-8) (300mg, 1.0mmol) and 60% sodium hydride mineral oil dispersion (61mg, 1.1mmol) were dissolved in 0 ℃ anhydrous dimethylformamide (9ml) under nitrogen, and after stirring for 10 minutes, CH was added₂Cl₂4-chlorobenzyl bromide (265mg, 1.2mmol) in (1ml) was added dropwise and the reaction mixture was stirred at room temperature for 1 hour. The solvent was removed under reduced pressure and the residue was dissolved in ethyl acetate (25mol), washed with water (2X 25ml), dried (magnesium sulphate), filtered and evaporated under reduced pressure. The residue was purified using flash chromatography eluting with 4: 1 heptane to ethyl acetate solvent to give the title compound as an oil (170mg, 40% yield).

¹H-NMR(400MHz，CDCl₃)δ＝1.4-1.5(m，9H)，2.0-2.45(m，2H)，3.5-3.8(m，5H)，4.05-4.2(s，1H)，4.25-4.4(m，1H)，4.4-4.55(m，2H)，7.3(m，4H).

LCMS (electrospray): m/z [ MNa ]⁺]392.

Preparation 4

(2S, 4S) -1- (tert-butoxycarbonyl) -4- [ (4-chlorobenzyl) oxy]-pyrrolidine-2-carboxylic acid

The title compound from preparation 3 (157mg, 0)42mmol) was dissolved in tetrahydrofuran (10 ml). Reacting LiOH₂O (54mg, 1.3mmol) was dissolved in water (5ml), and the two solutions were mixed, stirred at room temperature for 2 days, and then evaporated to dryness under reduced pressure. The residual residue was dissolved in ethyl acetate (25ml) and washed with saturated citric acid (25 mol). The organic portion was dried (magnesium sulfate), filtered and evaporated to dryness under reduced pressure. Purification using flash chromatography eluting with a solvent fraction of 20: 1 dichloromethane: methanol gave the title compound as an oil (106mg, 71% yield).

¹H-NMR(400MHz，CDCl₃)δ＝1.4(m，9H)，2.9-3.0(m，1H)，3.4-3.6(m，2H)，4.2-4.7(m，5H)，7.2-7.35(m，4H).

LCMS (electrospray): m/z [ M ]^-]354

Preparation 5

(2S, 4S) -4- (4-bromo-phenylthio) -pyrrolidine-1, 2-dicarboxylic acid 1-tert-butyl ester 2-ethyl ester

Sodium ethoxide (112mg, 1.65mmol) was slowly added to a stirred solution of 4-bromophenylthiophenol (302mg, 1.65mmol) in EtOH (6ml) at room temperature under a nitrogen atmosphere. After 30 minutes a solution of (2S, 4R) -4- (toluene-4-sulfonyloxy) -pyrrolidine-1, 2-dicarboxylic acid 1-tert-butyl ester 2-methyl ester (CASReg. No.88043-21-4) (300mg, 0.75mmol) in 1ml EtOH was added and the solution stirred for 48 hours. The reaction mixture was poured into 0.5M NaOH (50ml) and CH was used₂Cl₂(2X 50ml) was extracted. The combined organics were dried (magnesium sulfate) and concentrated in vacuo. Flash column chromatography gave the product as a pink solid (120mg, 40%).

¹H-NMR(400MHz，CDCl₃)δ＝1.25(3H，t)，1.40(9H，s)，2.00(1H，s)，2.60(1H，m)，3.35(1H，m)，3.60(1H，m)，3.90(1H，s)，4.18(2H，q)，4.22(1H，m)，7.35(2H，d)，7.40(2H，d).

LRMS (electrospray): m/z [ MNa ]⁺]454.

Preparation 6

(2S, 4S) -4- (phenylthio) -pyrrolidine-1, 2-dicarboxylic acid 1-tert-butyl ester 2-ethyl ester

The title compound was prepared by the method of preparation 5 in 40% yield as a pink solid.

¹H-NMR(400MHz，CDCl₃)δ＝1.23(3H，t)，1.41(9H，s)，2.00(1H，m)，2.61(1H，m)，3.38(1H，m)，3.62(1H，m)，3.90-4.03(1H，m)，4.15-4.35(3H，m)，7.20-7.50(5H，m).

LRMS (electrospray): m/z [ MNa ]⁺]374.

Preparation 7

(2S, 4S) -4- (4-bromo-phenylthio) -pyrrolidine-1, 2-dicarboxylic acid 1-tert-butyl ester

(2S, 4S) -4- (4-bromo-phenylthio) -pyrrolidine-1, 2-dicarboxylic acid 1-tert-butyl ester 2-ethyl ester (preparation 5, 120mg, 0.30mmol) was dissolved in MeOH (6ml) and 2M sodium hydroxide (0.83ml, 1.66mmol) was added. The solution was stirred for 14 h, concentrated and added to 0.5M HCl (50 ml). By CH₂Cl₂The aqueous layer was extracted (50ml), dried and concentrated. Flash column chromatography (eluting first with CH2Cl2 and then with 95% CH2Cl 2/MeOH) afforded the acid as a clear liquid (130mg, 48%).

¹H-NMR(400MHz，CDCl₃)δ1.43(9H，s)，2.4-2.8(2H，m)，3.35(1H，m)，3.62(1H，m)，3.8-4.0(1H，m)，4.3-4.4(1H，m)，7.28(2H，m)，7.41(2H，m).

LRMS (electrospray): m/z [ M ]^-]400，402.

Preparation 8

(2S, 4S) -4- (phenylthio) -pyrrolidine-1, 2-dicarboxylic acid 1-tert-butyl ester

The title compound was prepared by the method of preparation 7 from the title compound of preparation 6 in 83% yield as a clear oil.

¹H-NMR(400MHz，CDCl₃)δ1.41(9H，s)，2.10(0.5H，m)，2.38(0.5H，m)，2.50-2.75(1H，m)，3.36(1H，m)，3.62(1H，m)，3.82-4.03(1H，m)，4.25-4.41(1H，m)，7.20-7.45(5H，m).

LRMS (electrospray): m/z [ M ]^-]322.

Preparation 9

4- (2-fluoro-phenoxy) -pyrrolidine-1, 2-dicarboxylic acid 1-tert-butyl ester 2-methyl ester

(2S, 4R) -4-hydroxy-pyrrolidine-1, 2-dicarboxylic acid 1-tert-butyl ester 2-methyl ester (CAS Reg.No.74844-91-0) (300mg, 1.22mmol) was dissolved in THF (10ml), triphenylphosphine (385mg, 1.47mmol) and 2-fluorophenol (164.5mg, 1.47mmol) were added. The reaction was cooled in ice, DIAD (0.23ml, 1.2mmol) was added dropwise, and the reaction mixture was cooled at room temperatureThe reaction was stirred overnight. The mixture was concentrated in vacuo and CH was added₂Cl₂(20ml) and washed with 2N NaOH (10 ml). The phases were separated and washed with saturated brine (10ml), dried over MgSO4 and evaporated. The residue was dissolved in a minimum amount of diethyl ether and pentane was added until just the solution remained. After seeding (seeding) with triphenylphosphine oxide, the solution was cooled in ice and the resulting precipitate was filtered. The filtrate was evaporated and the residue was purified by flash chromatography on silica gel (50g), eluting first with pentane: diethyl ether (2: 1 vol) and then pentane: diethyl ether (1: 1 vol) to give the title product as an impure oil containing diisopropyldicarbamate as an impurity (388mg, 58%).

¹H-NMR(400MHz，CDCl₃)：δ＝1.45(d，9H)，2.35-2.57(m，2H)，3.65-3.79(m，5H)，4.43-4.57(m，1H)，4.88-5.02(m，1H)，6.81-6.98(m，2H)，6.98-7.10(m，2H).

LRMS (electrospray): m/z [ MNa ]⁺]362

Preparation 10

(2S, 4S) -4- (2-fluoro-phenoxy) -pyrrolidine-1, 2-dicarboxylic acid 1-tert-butyl ester

The ester from preparation 9 (400mg, 1.18mmol) was dissolved in THF (4ml) and LiOH. H in water (2ml) was added₂O (106mg, 3.53 mmol). The mixture was stirred at room temperature overnight. In use of CH₂Cl₂After washing (10ml), the aqueous solution was adjusted to pH 2 with saturated aqueous citric acid and CH₂Cl₂(2X 10ml) was re-extracted. The combined organic extracts were back-washed with saturated brine, dried over MgSO4, filtered and evaporated to give the title compound as a white solid (383mg, 49%) which was found by NMR to contain a small amount of diisopropyl dicarbamate (2%) as an impurity.

¹H-NMR(400MHz，CDCl₃)：1.16-1.70(m，9H)，2.20-2.92(m，2H)，3.58-3.85(m，2H)，4.38-4.63(m，1H)，4.83-5.02(m，1H)，6.78-7.17(m，4H).

LRMS (electrospray): m/z [ M-1]324

Preparation 11

(2S, 4S) -4- (4-chloro-phenoxy) -pyrrolidine-1, 2-dicarboxylic acid 1-tert-butyl ester 2-methyl ester

(2S, 4R) -4-hydroxy-pyrrolidine-1, 2-dicarboxylic acid 1-tert-butyl ester 2-methyl ester (CAS Reg.No.74844-91-0) (1.10g, 4.08mmol) was dissolved in THF (25ml), and 4-chlorophenol (0.78g, 6.12mmol) and triphenylphosphine (1.6g, 6.12mmol) were added. The solution was cooled in an ice bath and DIAD (0.96ml, 4.88mmol) was added dropwise. The reaction was stirred at room temperature overnight. After evaporation of the solvent, the residue was dissolved in diethyl ether (20ml) and pentane was added until the solution was just maintained. Seeding with triphenylphosphine oxide and cooling the solution in ice. The resulting precipitate was filtered and the filtrate was evaporated. The residue was purified by flash chromatography on silica gel (100g), the column was packed with pentane: diethyl ether (2: 1 vol), and eluted with pentane: diethyl ether (1: 1 vol) to give the title compound as a colorless oil (1.35g, 69%) which was found by NMR to contain a small amount of diisopropyl dicarbamate (CAS Reg. No.19740-72-8) as an impurity.

¹H-NMR(400MHz，CDCl₃)：δ＝1.43(d，9H)，2.36-2.57(m，2H)，3.61-3.81(m，5H)，4.39-4.59(m，1H)，4.80-4.90(m，1H)，6.64-6.78(m，2H)，7.18-7.30(m，2H).

LRMS (electrospray): m/z [ MNa ]⁺]378

Preparation 12

(2S, 4S) -4- (4-chloro-phenoxy) -pyrrolidine-1, 2-dicarboxylic acid 1-tert-butyl ester

The ester from preparation 11 was dissolved in THF (30ml) and LiOH₂A solution of O (440mg, 10.56mmol) in water (15 ml). The reaction was stirred at room temperature overnight and then the solvent was evaporated in vacuo. In CH₂Cl₂The residue was partitioned between (20ml) and saturated aqueous citric acid solution (10ml) and the phases separated. The organic layer was washed with saturated brine (10ml), dried over MgSO4, and evaporated. The crude product was partially purified by flash chromatography on silica gel (100g), first with CH₂Cl₂Eluting, then with CH₂Cl₂MeOH (25: 1 by volume) gave a material which was found by NMR to still contain diisopropyldicarbamate. Recrystallization from EtOAc provided white crystals which were filtered and washed with EtOAc: pentane (1: 1) to provide the title compound (517mg, 55%).

¹H-NMR(400MHz，CDCl₃)：δ＝1.23-1.67(m，9H)，2.20-2.88(m，2H)，3.55-3.81(m，2H)，4,40-4.61(m，1H)，4.78-4.92(m，1H)，6.63-6.84(m，2H)，7.11-7.32(m，2H)

LRMS (electrospray): m/z [ M-1]340

Preparation 13

(2S, 4S) -4- (isoquinoline-7-yloxy) -pyrrolidine-1, 2-dicarboxylic acid di-tert-butyl ester

The title compound was synthesized from (2S, 4R) -4-hydroxy-pyrrolidine-1, 2-dicarboxylic acid di-tert-butyl ester (CAS reg.no.170850-75-6) and isoquinolin-7-ol using the same method as preparation 11 and was obtained as an oil in 15% yield.

¹H-NMR(400MHz，CDCl₃)：δ＝1.41-1.53(m，18H)，2.43-2.63(m，2H)，3.68-3.97(m，2H)，4.30-4.52(m，1H)，4.99-5.06(m，1H)，7.08-7.16(m，1H)，7.41-7.77(m，，3H)，8.42(d，1H)，9.10-9.18(m，1H).

LCMS (electrospray): m/z [ MH⁺]415

Preparation 14

(2S, 4S) -4- (3-chloro-phenoxy) -pyrrolidine-1, 2-dicarboxylic acid 1-tert-butyl ester 2-methyl ester

In N₂Next, diisopropyl azodicarboxylate (0.29ml, 1.47mmol) was added dropwise to a stirred solution of (2S, 4R) -4-hydroxy-pyrrolidine-1, 2-dicarboxylic acid 1-tert-butyl ester 2-methyl ester (CAS Reg.No.74844-91-0) (0.3g, 1.22mmol), 3-chlorophenol (0.189g, 1.47mmol) and triphenylphosphine (0.385g, 1.47mmol) in THF (2ml) cooled at 0 ℃. The mixture was stirred at room temperature for 3 days. The solvent was removed in vacuo and the product was purified by chromatography on silica gel using ether/n-pentane: 40/60 As eluent, 0.27g (62%) of the title compound and reduced diisopropyl azodicarboxylate (1/1) mixture as oil were obtained.

¹H NMR(400MHz，CDCl₃)：δ＝1.46，1.49(2×s，9H)，2.47(2H，m)，3.71(5H，m)，4.42(1H，m)，4.42，4.54(1H，2×m)，4.87(1H，m)，6.68(1H，m)，6.79(1H，s)，6.92(1H，m)，7.18(1H，m).

LRMS (electrospray): m/z 378 (MNa)⁺).

Preparation 15

(2S, 4S) -4- (3-chloro-phenoxy) -pyrrolidine-1, 2-dicarboxylic acid 1-tert-butyl ester

To the product from preparation 14(0.25g, 0.7mmol) in THF (4ml) was added a solution of lithium hydroxide (50mg) in water (4 ml). The mixture was stirred overnight, then water (10ml) and diethyl ether (20ml) were added. The aqueous phase was washed twice with diethyl ether (2X 20ml) and acidified with 2N HCl and extracted with diethyl ether (2X 20 ml). The ether phase was dried (magnesium sulfate), filtered and evaporated to give the title compound (80mg, 33%).

¹H NMR(400MHz，CDCl₃)：δ＝1.42，1.48(2×s，9H)，2.30-2.70(m，2H)，3.60-3.80(m，2H)，4.40-4.60(m，1H)，4.86(m，1H)，6.71(m，1H)，6.82(m，1H)，6.94(m，1H)，7.16(m，1H).

LRMS (electrospray): m/z [ MNa ]⁺]364，340[M-1]340.

Preparation 16

(2S, 4S) -4-benzyloxy-pyrrolidine-1, 2-dicarboxylic acid 1-tert-butyl ester 2-methyl ester

Under nitrogen atmosphere at 0 deg.C, (2S, 4)S) -4-hydroxy-pyrrolidine-1, 2-dicarboxylic acid 1-tert-butyl ester 2-methyl ester (CAS Reg.No.227935-38-8) (300mg, 1.2mmol) and 60% sodium hydride mineral oil dispersion (61mg, 1.5mmol) were dissolved in anhydrous dimethylformamide (9 ml). After stirring for 10 minutes, add dropwise to CH₂Cl₂Benzyl bromide (0.153ml, 1.3mmol) in (1ml) the reaction mixture was stirred at room temperature for 1 hour. The solvent was removed under reduced pressure and the residue was dissolved in ethyl acetate (25ml) and washed with water (2X 25mol), dried (magnesium sulphate), filtered and evaporated under reduced pressure. The residue was purified by flash chromatography eluting with a solvent gradient of 4: 1 heptane: ethyl acetate to give the title compound (167mg, 42% yield) as an oil.

¹H-NMR(400MHz，CDCl₃)δ＝1.2-1.6(m，12H)，2.2-2.45(m，1H)，3.4-3.8(m，4H)，4.05-4.2(m，1H)，4.3-4.5(m，2H)，7.15-7.4(m，5H).

LCMS (electrospray): m/z [ MNa ]⁺]358.

Preparation 17

(2S, 4S) -1- (tert-Butoxycarbonyl) -4- (benzyloxy) -pyrrolidine-2-carboxylic acid

The title compound from preparation 16(167mg, 0.5mmol) was dissolved in tetrahydrofuran (10 ml). LIOH.H₂O (63mg, 1.5mmol) was dissolved in water (5 ml). The two solutions were mixed, stirred at room temperature for two days and then evaporated to dryness under reduced pressure. The residual residue was dissolved in ethyl acetate (25ml) and washed with saturated citric acid (25 ml). The organic portion was dried (magnesium sulfate), filtered and evaporated to dryness under reduced pressure. The crude compound (150mg, 94% yield) was removed as an oil and used in the next stage (example 9).

LCMS (electrospray): m/z [ M ]^-]320，[MNa⁺]344.

Preparation 18

(2S, 4S) -4- (2, 3-dihydro-benzofuran-6-yloxy) -pyrrolidine-1, 2-dicarboxylic acid 1-tert-butyl ester 2-methyl ester

The title compound was prepared as a white solid in 41.6% yield from (2S, 4R) -4-hydroxy-pyrrolidine-1, 2-dicarboxylic acid 1-tert-butyl-ester 2-methyl ester and 2, 3-dihydro-benzofuran-6-ol using the procedure of preparation 14.

¹H-NMR(400MHz，CDCl₃)：δ＝1.43(d，9H)；2.36-2.50(m，2H)；3.03-3.17(m，2H)；3.59-3.80(m，5H)；4.15-4.41(m，3H)；4.78-4.83(m，1H)；6.21-6.32(m，2H)；6.98-7.02M，1H).

LRMS (electrospray): [ MNa)⁺]386

Preparation 19

(2S, 4S) -4- (2, 3-dihydro-benzofuran-6-yloxy) -pyrrolidine-1, 2-dicarboxylic acid 1-tert-butyl ester

From 4- (2, 3-dihydro-benzofuran-6-oxy) -pyrrolidine-1, 2-dicarboxylic acid 1-tert-butyl ester 2-methyl ester the title compound was prepared by the method of preparation 15 as a white solid in 78% yield.

¹H-NMR(400MHz，CDCl₃)：δ＝1.38-1.58(m，9H)；2.21-2.83(m，2H)；3.02-3.18(m，2H)；3.59-3.82(m，2H)；4.38-4.60(m，3H)；4.80-4.90(m，1H)；6.22-6.42(m，2H)；6.97-7.10(m，1H).

LRMS (electrospray): [ M-1]348

Preparation 20

4- (3-fluoro-benzylidene) -pyrrolidine-1, 2-dicarboxylic acid 1-tert-butyl ester 2-methyl ester

At room temperature, m-fluorobenzyltriphenylphosphonium²To a solution of bromide (8.08g, 0.018mmol) in anhydrous dichloromethane (200ml) was added potassium tert-butoxide (1M in THF, 17.2ml, 0.017mmol) dropwise and stirred for 1 hour. The mixture was cooled to 0 ℃ and to this solution was added dropwise (2S) 4-oxo-pyrrolidine-1, 2-dicarboxylic acid 1-tert-butyl ester 2-methyl ester in dichloromethane (20ml)³(3.8g, 0.016 mmol). The mixture was warmed to room temperature and stirred for 18 hours. The reaction was quenched with saturated ammonium chloride (100ml), the aqueous layer was extracted with dichloromethane (2X 100ml), and the combined organics were dried over magnesium sulfate. The solvent was evaporated under reduced pressure. The residue was purified by flash chromatography on silica gel eluting with a solvent gradient of heptane to ethyl acetate (4: 1) to give the title compound (3.48g, 67%) as a colorless oil.

¹H-NMR(400MHz，CD₃OD) (mixture of cis and trans geometric isomers): δ is 1.44(s, 10H), 1.50(s, 8H), 2.79-2.94(m, 2H), 3.20-3.37(m, 2H), 3.66(d, 3H), 3.72(d, 3H), 4.20-4.38(m, 4H), 4.42-4.48(m, 1H), 4.52-4.60(m, 1H), 6.42-6.51(m, 2H), 6.89-7.10(m, 6H), 7.30-7.40(m, 2H).

LRMS(APCI)：m/z[(M+H)-Boc]⁺236.

Microanalysis: the following are found: c, 64.46; h, 6.77; n, 4.07%. C₁₈H₂₂FNO₄C, 64.46; h, 6.61; n, 4.18 percent.

K.rafizadeh and k.yates; chem.1984, 49, 9, 1500-.

3.Org.Lett，2001，3041-3043.

Preparation of 21 to 24

By a method similar to preparation 20, using the appropriate phosphonium bromide salt and (2S) 4-oxo-pyrrolidine-1, 2-dicarboxylic acid 1-tert-butyl ester 2-methyl ester³Examples of the formulae listed below were prepared:

preparation 25

4- (3-fluoro-benzylidene) -pyrrolidine-1, 2-dicarboxylic acid 1-tert-butyl ester

To a stirred solution of 1-tert-butyl 2-methyl 4- (3-fluoro-benzylidene) -pyrrolidine-1, 2-dicarboxylate (3.23g, 9.63mmol) in tetrahydrofuran (150ml) was added 1M lithium hydroxide monohydrate (1.21g, 28.9mmol) in water (50 ml). The mixture was stirred at room temperature for 3 days. The tetrahydrofuran was evaporated under reduced pressure, the residue diluted with water (30ml) and acidified to pH 2.0-3.0 using 1M hydrochloric acid. The water was extracted with diethyl ether (3X 100ml) and the combined organics were dried over magnesium sulphate. The solvent was evaporated under reduced pressure to give the title compound (2.37g, 77%) as a white foam.

¹H-NMR(400MHz，CD₃OD) (cis and trans geometric isomersMixture of bodies): δ is 1.44(s, 5H), 1.50(s, 4H), 2.80-2.96(m, 1H), 3.20-3.38(m, 1H), 4.24-4.34(m, 2H), 4.45-4.45(m, 0.5H), 4.46-4.58(m, 0.5H), 6.43-6.54(m, 1H), 6.90-7.05(m, 3H), 7.30-7.40(m, 1H).

LRMS(APCI)：m/z[M-H]⁺320.

Microanalysis: the following are found: c, 63.10; h, 6.53; n, 4.05%. C₁₇H₂₀NO₄F. C, 63.54; h, 6.27; n, 4.36 percent.

Preparation of 26 to 29

Examples of the general formulae listed below were prepared by a method similar to preparation 25, using the appropriate starting ester:

preparation 30

4- (3-fluoro-benzylidene) -pyrrolidine-1, 2-dicarboxylic acid 1-tert-butyl ester 2- (2-isopropyl-5-methyl-cyclo Hexyl) ester

To a solution of 4- (3-fluoro-benzylidene) -pyrrolidine-1, 2-dicarboxylic acid 1-tert-butyl ester (2.68g, 8.35mmol) was added 1R, 2S, 5R (-) menthol (1.31g, 8.35mmol) followed by dimethylaminopyridine (1.02g, 8.35 mmol). The mixture was cooled to 0 ℃ and dicyclohexylcarbodiimide (1.89g, 9.19mmol) in dichloromethane (10ml) was added in one portion. The mixture was warmed to room temperature and stirred for 18 hours. The mixture was filtered and the filtrate was washed with 1N hydrochloric acid (30ml), sodium bicarbonate salt (30ml) and water (30 ml). The organics were dried over magnesium sulfate and the solvent was evaporated under reduced pressure. flash master column chromatography eluting with heptane: ethyl acetate (12: 1) gave the title compound (1.20g, 31%) as a colorless oil.

¹H-NMR(400MHz，CD₃OD)：δ＝0.55(t，2H)，0.69(t，2H)，0.80-0.93(m，8H)，0.95-1.05(m，1H)，1.20-1.35(m，2H)，1.44(d，9H)，1.60-2.00(m，3H)，2.73-2.90(m，1H)，4.03-4.68(m，4H)，6.43-6.52(m，1H)，6.93-7.11(m，3H)，7.33-7.40(m，1H).

LRMS(APCI)：m/z[MH]⁺460.

Preparation of 31-34

Examples of the general formulae listed below were prepared by a method analogous to preparation 30, using the appropriate starting acid:

preparation 35

4- (3-fluoro-benzyl) -pyrrolidine-1, 2-dicarboxylic acid 1-tert-butyl ester 2- (2-isopropyl-5-methyl-cyclohexane) Yl) ester.

4- (3-fluoro-benzylidene) -pyrrolidine-1, 2-dicarboxylic acid 1-tert-butyl ester 2- (2-isopropyl-5-methyl-cyclohexyl) ester (1.20g, 2.61mmol) was dissolved in ethyl acetate: toluene (1: 1, 12 ml). The solution was hydrogenated over platinum oxide (120mg, 10% by weight) at 25 deg.C and 15psi for 1 hour. The reaction mixture was filtered through arbocel and the filtrate was concentrated under reduced pressure. The residue was purified by flash master chromatography eluting with heptane to ethyl acetate (15: 1) to give the title compound as a colorless oil (1.11g, 91%).

¹H-NMR(400MHz，CD₃OD)：δ＝0.72-1.37(m，13H)，1.44(d，9H)，1.43-1.75(m，4H)，1.87-2.01(m，2H)，2.31-2.58(m，2H)，2.83(d，2H)，3.07(t，1H)，3.50-3.65(m，1H)，4.13-4.30(dt，1H)，4.71(td，1H)，6.90(d，2H)，7.00(d，1H)，7.30(q，1H).

LRMS(APCI )：m/z[MH-BOC]⁺362.

Preparation 36 to 39

Examples of the general formulae listed below were prepared by a method analogous to preparation 35, using the appropriate starting material, the olefinic menthyl ester:

supplementary notes

1. Hydrogenation using rhodium on alumina (5%) (44mg, 10% by weight) at 50 deg.C, 70psi produced the title compound of 33 for 24 hours.

Preparation 40

(2S, 4S) -4- (3-chloro-phenylamino) -pyrrolidine-1, 2-dicarboxylic acid 1-tert-butyl ester 2-methyl ester

4-oxo-pyrrolidine-1, 2-dicarboxylic acid 1-tert-butyl ester 2-methyl ester (364.5mg, 1.5mmol) and 3-chloroaniline (191mg, 1.5mmol) were dissolved in DCM (10 ml). To the solution was added sodium triacetoxyborohydride (413mg, 1.95mmol) and acetic acid (0.085ml, 1.5mmol), and the reaction was stirred at room temperature overnight. The reaction mixture was washed with 2N NaOH (5ml), saturated brine (5ml), and MgSO₄Dried and evaporated. The residue was purified by flash chromatography on silica gel eluting with DCM to give the title compound as a colourless oil (215mg, 40%).

¹H-NMR(400MHz，CDCl₃)：δ＝1.42(d，9H)；2.04-2.17(m，1H)；2.39-2.55(m，1H)；3.48-3.61(m，1H)；3.63-3.79(m，4H)；4.02-4.15(m，1H)；4.25-4.41(m，1H)；6.42-6.51(m，1H)；6.55-6.61(m，1H)；6.65-6.75(m，1H)；7.01-7.11(m，1H).

LRMS (electrospray): [ MNa)⁺]377.

Preparation 41

(2S, 4S) -4- (3-chloro-phenylamino) -pyrrolidine-1, 2-dicarboxylic acid 1-tert-butyl ester

To a solution of (2S, 4S) -4- (3-chloro-phenylamino) -pyrrolidine-1, 2-dicarboxylic acid 1-tert-butyl ester 2-methyl ester (200mg, 0.58mmol) in THF (2ml) was added a solution of lioh.h2o (73mg, 1.74mmol), and the reaction was stirred at room temperature overnight. The solvent was concentrated in vacuo and the remaining aqueous solution was washed with DCM (2 ml). The aqueous layer was adjusted to pH 5 with saturated aqueous citric acid and re-extracted with DCM (2X 10 ml). The combined extracts were dried with MgSO4 to give the title compound as a white foam (168mg, 88%).

¹H-NMR(400MHz，CDCl₃)：δ＝1.18-1.69(m，9H)；2.11-2.45(m，1H)；2.53-2.61(m，1H)；3.44-3.62(m，2H)；4.04-4.11(m，1H)；4.48-4.53(m，1H)；6.38-6.61(m，2H)；6.65-6.74(m，1H)；7.04-7.15(m，1H).

LRMS (electrospray): [ M-1]339

Preparation 42

4-hydroxymethyl-pyrrolidine-1, 2-dicarboxylic acid di-tert-butyl ester

To a solution of 2-methyl-2-butene (2M in tetrahydrofuran, 30ml, 60mmol) in anhydrous tetrahydrofuran (40ml) was added borane-tetrahydrofuran complex (1M in tetrahydrofuran, 30ml, 30mmol) dropwise under nitrogen atmosphere at 0 ℃ over 10 minutes and stirring was maintained for 2 hours. The reaction mixture was cooled to-20 ℃, a solution of di-tert-butyl 4-methylene-pyrrolidine-1, 2-dicarboxylate (2.84g, 10mmol) (CAS reg 163190-46-3) tetrahydrofuran (20ml) was added dropwise and stirred at room temperature for 18 hours. Water (40mol) was added carefully followed by sodium hydroxide (0.5M, 20ml) and then hydrogen peroxide (27.5% w/w in water, 10mol) and stirred at room temperature for 2 hours. The organic solvent was removed under reduced pressure, and the aqueous layer was extracted with ethyl acetate (2X 60 ml). The combined extracts were dried (MgSO4), filtered and evaporated under reduced pressure. The residue was purified by chromatography on silica gel eluting with 40% ethyl acetate/heptane to give the title compound as a colorless oil, a mixture of diastereomers (. about.5: 12S, 4S: 2S, 4R) (1.25g, 41%).

¹H-NMR(400MHz，CD₃OD)：δ＝1.39-1.49(m，18H)；1.63-1.75(m，0.8H)；1.96-2.07(m，0.4H)；2.32-2.47(m，1.8H)；3.11-3.20(m，1H)；3.46-3.53(m，2H)；3.53-3.60(m，0.2H)；3.60-3.68(m，0.8H)；4.09-4.2(m，1H)

LRMS (electrospray): [ M +23] 324; [ M-1]300

Preparation 43

4- (3-fluoro-phenoxymethyl) -pyrroline-1, 2-dicarboxylic acid di-tert-butyl ester

To a solution of di-tert-butyl 4-hydroxymethyl-pyrrolidine-1, 2-dicarboxylate (preparation 42, 500mg, 1.66mmol), triphenylphosphine (653mg, 2.49mmol) and 3-fluorophenol (0.23ml, 2.49mmol) in tetrahydrofuran (30ml) was added dropwise diisopropyl azodicarboxylate (0.49ml, 2.49mmol) at 0 ℃ under a nitrogen atmosphere over 5 minutes, and stirred at room temperature for 72 hours. The solvent was removed under reduced pressure and the residue was purified by chromatography on silica gel eluting with 10-15% ethyl acetate/heptane to give the title compound as a colorless oil, a mixture of diastereomers (. about.5: 12S, 4S: 2S, 4R).

¹H-NMR(400MHz，CD₃OD)：δ＝1.39-1.49(m，18H)；1.81-1.95(m，0.8H)；2.09-2.20(m，0.4H)；2.44-2.59(m，0.8H)；2.65-2.80(m，1H)；3.22-3.33(m，1H)；3.65-3.75(m，1H)；3.91-4.00(m，1.8H)；4.00-4.07(m，0.2H)；4.14-4.26(m，1H)；6.60-6.74(m，3H)；7.20-7.28(m，1H)

LRMS (electrospray): [ M +23]418

Preparation 44

(2S, 4S) -pyrrolidine-1, 2, 4-tricarboxylic acid 1, 2-di-tert-butyl ester

4-phenyl-pyrrolidine-1, 2-dicarboxylic acid di-tert-butyl ester (CAS Reg. No. 344286-69-7) stirred at 0 ℃ under nitrogen⁵(0.78g, 2.24mmol) and sodium periodate (5.77g, 27mmol) to a mixture of ethyl acetate (5.5mol), acetonitrile (5.5mol) and water (8.5ml) was added trichlororhodium (10mg, 0.05mmol) and stirred at room temperature for 18 hours. Diethyl ether (20ml) was added and stirred for a further 1 hour. 1M hydrochloric acid (5ml) was added to the solution, and the mixture was extracted with ethyl acetate (3X 30 ml). The organic extracts were combined, dried (MgSO4), filtered and evaporated under reduced pressure. The residue was purified by chromatography on silica gel eluting with 50: 1 ethyl acetate: heptane: glacial acetic acid to give the title compound as a colourless gum (501mg, 78%).

¹H-NMR(400MHz，CDCl₃)：δ＝1.40-1.49(m，18H)；2.26-2.40(m，1H)；2.42-2.56(m，1H)；3.02-3.12(m，1H)；3.65-3.80(m，1.4H)& 3.80-3.88(m，0.6H)[rotamers]；4.09-4.20(m，0.7H)& 4.20-4.26(m，0.3H)[rotamers]

LRMS (electrospray): [ M-1]314

5J.Org.Chem.，2001，3593-3596

Preparation 45

(2S, 4S) -4-hydroxymethyl-pyrrolidine-1, 2-dicarboxylic acid di-tert-butyl ester

To a solution of pyrrolidine-1, 2, 4-tricarboxylic acid 1, 2-di-tert-butyl ester (preparation 44, 501mg, 1.59mmol) in anhydrous tetrahydrofuran (10ml) was added borane-tetrahydrofuran complex (1M in tetrahydrofuran, 3.16ml, 3.18mmol) dropwise at 0 ℃ under a nitrogen atmosphere, and stirred at room temperature for 18 hours. The solvent was removed under reduced pressure and the residue was dissolved in ethyl acetate (10ml) and washed with 1M hydrochloric acid (10ml), saturated sodium bicarbonate (10ml), then dried (MgSO4), filtered and evaporated under reduced pressure to give the title compound as a colourless gum (132 mg, 27% of the single diastereoisomer).

¹H-NMR(400MHz，CDCl₃)：δ＝1.40-1.47(m，18H)；1.59-1.80(m，1H)；1.80-2.00(m，1H)；2.31-2.46(m，2H)；3.14-3.21(m，1H)；3.54-3.65(m，2H)；3.65-3.74(m，1H)；4.10-4.20(m，1H).

Preparation 46

(2S, 4S) -4- (3-chloro-phenoxymethyl) -pyrrolidine-1, 2-dicarboxylic acid di-tert-butyl ester

To a solution of di-tert-butyl 4-hydroxymethyl-pyrrolidine-1, 2-dicarboxylate (preparation 45, 132mg, 0.44mmol), triphenylphosphine (172mg, 0.66mmol) and 3-chlorophenol (0.069ml, 0.66mmol) in tetrahydrofuran (5ml) was added dropwise diisopropyl azodicarboxylate (0.129 ml; 0.66mmol) at 0 ℃ under a nitrogen atmosphere, and stirred at room temperature for 18 hours. The solvent was removed under reduced pressure and the residue was purified by chromatography on silica gel eluting with 10% ethyl acetate/heptane to give the title compound as a colourless gum (66mg, 37%).

¹H-NMR(400MHz，CDCl₃)：δ＝1.40-1.56(m，18H)；1.80-1.91(m，1H)；2.40-2.54(m，1H)；2.61-2.70(m，1H)；3.24-3.33(m，1H)；3.67-3.74(m，0.3H)& 3.74-3.81(m，0.7H)[rotamers]；3.84-3.96(m，2H)；4.12-4.20(m，0.7H)& 4.20-4.26(m，0.3H)[rotamers]；6.67-6.75(m，1H)；6.82-6.86(m，1H)；6.86-6.93(m，1H)；7.10-7.19(m，1H)

LRMS (electrospray): [ M +23]434

Pharmaceutical composition examples

In the following examples, the term "active compound" or "active ingredient" refers to a compound of formula (I) according to the present invention or a pharmaceutically acceptable salt, solvate, polymorph or prodrug thereof.

(i)Tablet composition

The following compositions a and B can be prepared by wet granulation of the ingredients of (a) - (c) and (a) - (d) with a solution of povidone, followed by addition of magnesium stearate and compression.

Composition A

mg/tablet mg/tablet

(a) Active ingredient 250250

(b) Lactose b.p. 21026

(c) Sodium starch glycolate 2012

(d) Povidone B.P.159

(e) Magnesium stearate5 3

500 300

Composition B

mg/tablet

(a) Active ingredient 250250

(b) Lactose 150150-

(c)Avicel PH 101 60 26

(d) Sodium starch glycolate 2012

(e) Povidone B.P.159

(f) Magnesium stearate5 3

500 300

Composition C

mg/tablet

Active ingredient 100

Lactose 200

Starch 50

Povidone 5

Magnesium stearate4

359

The following compositions D and E can be prepared by direct compression of the mixed ingredients. The lactose used in formulation E was of the direct compression type.

Composition D

mg/tablet

Active ingredient 250

Magnesium stearate 4

Pregelatinized starch NF 15146

400

Composition E

mg/tablet

Active ingredient 250

Magnesium stearate 5

Lactose 145

Avicel 100

500

Composition F (controlled Release composition)

mg/tablet

(a) Active ingredient 500

(b) Hydroxypropyl methylcellulose 112

(Methocel K4M Premium)

(c) Lactose B.P. 53

(d) Povidone B.P.C.28

(e) Magnesium stearate 7

700

The composition may be prepared by wet granulation of ingredients (a) to (c) with a solution of povidone, followed by addition of magnesium stearate and compression.

Composition G (enteric coated tablets)

Enteric-coated tablets of composition C may be prepared by coating the tablets with 25 mg/tablet of an enteric polymer such as cellulose acetate phthalate, polyvinyl acetate phthalate, hydroxypropyl methylcellulose phthalate or an anionic polymer of methacrylic acid or methyl methacrylate (Eudragit L). In addition to Eudragit L, these polymers should also include 10% (relative to the weight of polymer used) of a plasticizer to prevent film cracking during application or storage. Suitable plasticizers include diethyl phthalate, tributyl citrate, and triacetin.

Composition H (enteric coated controlled release tablets)

Enteric-coated tablets of composition F may be prepared by coating the tablets with 50 mg/tablet of an enteric polymer such as cellulose acetate phthalate, polyvinyl acetate phthalate, hydroxypropylmethylcellulose phthalate or an anionic polymer of methacrylic acid or methyl methacrylate (Eudragit L). In addition to Eudragit L, these polymers should also include 10% (relative to the weight of polymer used) of a plasticizer to prevent film cracking during application or storage. Suitable plasticizers include diethyl phthalate, tributyl citrate, and triacetin.

(ii)Capsule composition

Composition A

Capsules can be prepared by mixing the ingredients of composition D above and filling two-part (two-part) hard gelatin capsules with the resulting mixture. Composition B (below) can be prepared in a similar manner.

Composition B

mg/capsule

(a) Active ingredient 250

(b) Lactose B.P. 143

(c) Starch sodium glycolate 25

(d) Magnesium stearate 2

420

Composition C

mg/capsule

(a) Active ingredient 250

(b)Macrogol 4000 BP 350

600

Capsules may be prepared by melting Macrogol 4000 BP, dispersing the active ingredient in the melt and filling two-part hard gelatin capsules with it.

Composition D

mg/capsule

Active ingredient 250

Lecithin 100

Peanut oil 100

450

Capsules may be prepared by dispersing the active ingredient in lecithin and peanut oil and filling soft, elastic gelatin capsules with the dispersion.

Composition E (controlled release capsules)

mg/capsule

(a) Active ingredient 250

(b) Microcrystalline cellulose 125

(c) Lactose BP 125

(d) Ethyl cellulose 13

513

The controlled release capsule formulation can be prepared by extruding the mixed ingredients (a) to (c) using an extruder, followed by spheronization (spheronisation) and drying the extrudate. The dried pellets are coated with a controlled release film (d) and filled into two-part hard gelatin capsules.

Composition F (enteric capsule)

mg/capsule

(a) Active ingredient 250

(b) Microcrystalline cellulose 125

(c) Lactose BP 125

(d) Cellulose acetate phthalate 50

(e) Phthalic acid diethyl ester 5

555

The enteric capsule composition can be prepared by extruding the mixed ingredients (a) to (c) using an extruder, followed by spheronization and drying of the extrudate. The dried pellets are coated with an enteric film (d) containing a plasticizer (e) and filled into two-part hard gelatin capsules.

Composition G (enteric coated controlled release capsules)

Enteric capsules of composition E may be prepared by coating the tablets with 50 mg/capsule of an enteric polymer such as cellulose acetate phthalate, polyvinyl acetate phthalate, hydroxypropylmethylcellulose phthalate or an anionic polymer of methacrylic acid or methyl methacrylate (Eudragit L). In addition to Eudragit L, these polymers should also include 10% (relative to the weight of polymer used) of a plasticizer to prevent film cracking during application or storage. Suitable plasticizers include diethyl phthalate, tributyl citrate, and triacetin.

(iii) Intravenous injection composition

Active ingredient 0.200g

Sterile pyrogen-free phosphate buffer (pH 9.0) to 10ml

The active ingredient was dissolved in most phosphate buffer at 35-40 ℃ to make a volume and filtered through a sterile microporous filter into a sterile 10ml glass vial (type 1) sealed with a sterile lid and cover (overlals).

(iv) Intramuscular injection composition

Active ingredient 0.20g

Benzyl alcohol 0.10g

Glycofurol 75 1.45g

Proper amount of water for injection is 3.00ml

The active ingredient is dissolved in glycofurol. Then benzyl alcohol was added and dissolved, and water was added to 3 ml. The mixture was then filtered through a sterile microporous filter and sealed in a sterile 3ml glass vial (type 1).

(v) Syrup composition

Active ingredient 0.25g

Sorbitol solution 1.50g

Glycerol 1.00g

Sodium benzoate 0.005g

Flavoring agent 0.0125ml

Proper amount of pure water to 5.0ml

Sodium benzoate was dissolved in partially purified water and sorbitol solution was added. The active ingredient is added and dissolved. The resulting solution was mixed with glycerol and then adjusted to the desired volume with pure water.

(vi) Suppository composition

mg/suppository

Active ingredient 250

Hard Fat，BP(Witepsol H15-Dynamit NoBel) 1770

2020

1/5 of Witepsol H15 was melted in a steam jacketed vessel with a maximum temperature of 45 ℃. The active ingredient was sieved through a 2001m sieve and added to the molten matrix under stirring by using a Silverson equipped with a cutting head until a homogeneous dispersion was obtained. The mixture was maintained at 45 ℃ and residual Witepsol H15 was added to the suspension, which was stirred to ensure uniform mixing. The entire suspension was then cooled to 40 ℃ by passing it through a 2501m stainless steel mesh screen with continuous stirring. Portions of 2.02g of the mixture were filled into suitable plastic moulds at a temperature of 38-40 ℃ and the suppositories were allowed to cool to room temperature.

(vii) Vaginal suppository composition

mg/suppository

Active ingredient (631m) 250

Anhydrous glucose 380

Potato starch 363

Magnesium stearate 7

1000

The above ingredients are directly mixed, and suppositories are prepared by compressing the resulting mixture.

(viii) Transdermal composition

Active ingredient 200mg

Ethanol USP 0.1ml

Hydroxy ethyl cellulose

The active ingredient and ethanol USP were gelled with hydroxyethylcellulose. And filled in a container having a surface area of 10cm²In a transdermal device of (1).

In the above described assay, the compounds of the invention showed biological activity as shown in the following table.

Example No.2	IC50(nM)
Example No.2	IC50(nM)	2	119
4	72	2	119
4	72	7	210
8	5	7	210
8	5	10	11
11	15	10	11
11	15	12	7
14	9	12	7

Claims

1. Use of a compound of formula (I) or a pharmaceutically acceptable salt, solvate or prodrug thereof in the manufacture of a medicament for the treatment of neuropathic pain:

in the formula:

x is O, S, NH or CH₂And Y is CH₂Or a direct bond, or Y is O, S or NH and X is CH₂(ii) a And is

R is a 3-12 membered cycloalkyl, 2, 3-dihydrobenzofuranyl, aryl or heteroaryl group, wherein any ring may be optionally substituted with one or more substituents independently selected from: halogen, hydroxy, cyano, nitro, amino, hydroxycarbonyl, hydroxy-, cyano-, nitro-, amino-, or carboxyl group,

C₁-C₆Alkyl radical, C₁-C₆Alkenyl radical, C₁-C₆Alkynyl, alkynyl,

C₁-C₆Alkylthio radical, C₁-C₆Alkylthio carbonyl group, C₁-C₆An alkoxycarbonyl group, a carbonyl group,

3-8 membered cycloalkyl, 4-8 membered heterocycloalkyl, phenyl and monocyclic heteroaryl,

wherein said aryl is phenyl or naphthyl; the heteroaryl group is a mono-or bicyclic aromatic ring system containing at least one ring heteroatom independently selected from O, S and N.

2. Use according to claim 1, wherein R is cyclohexyl, 2, 3-dihydrobenzofuranyl, isoquinolinyl or phenyl, wherein said cyclohexyl, 2, 3-dihydrobenzofuranyl, isoquinolinyl or phenyl is optionally substituted with one or more substituents independently selected from:

halogen, hydroxy, cyano, nitro, amino, hydroxycarbonyl, hydroxy-, cyano-, nitro-, amino-, or carboxyl group,

C₁-C₆Alkyl radical, C₁-C₆Alkenyl radical, C₁-C₆Alkynyl, alkynyl,

3. Use according to claim 1 or 2, wherein R is phenyl optionally substituted with one or more substituents independently selected from the group consisting of:

C₁-C₆Alkyl radical, C₁-C₆Alkenyl radical, C₁-C₆Alkynyl, alkynyl,

4. Use according to any one of claims 1 to 3, wherein R is optionally selected from halogen, hydroxy and (C)₁-C₆) One or two of the alkoxy groups are substituted.

5. Use according to any one of claims 1 to 4 wherein R is substituted by one or two groups selected from methoxy, fluoro, chloro and bromo.

6. Use according to any one of claims 1 to 5, wherein X is O, S, NH or CH₂And Y is CH₂Or a direct bond, or X is CH₂And Y is O.

7. Use according to any one of claims 1 to 6, wherein-Y-X-is an oxy, thio, aminomethylene, methylenethio, methyleneoxy or oxymethylene linkage.

8. Use according to any one of claims 1 to 7, wherein-Y-X-is an oxy, methylene or oxymethylene linkage.

9. A compound of formula (Ia) or a pharmaceutically acceptable salt or solvate thereof,

wherein R is^aSelected from halogen, hydroxy, (C)₁-C₆) Alkoxy, cyano, nitro, amino, hydroxycarbonyl, amino, nitro, cyano, amino,

C₁-C₆alkyl radical, C₁-C₆Alkenyl radical, C₁-C₆Alkynyl, alkynyl,

Hydroxy radical C₁-C₆Alkyl radical, C₁-C₆Alkoxy radical C₁-C₆Alkyl, perfluoro C₁-C₆Alkyl, perfluoro C₁-C₆Alkoxy radical,

C₁-C₆Alkylthio carbonyl group, C₁-C₆An alkoxycarbonyl group, a carbonyl group,

C₁-C₆alkylsulfonyl radical, C₁-C₆An alkylsulfonylamino group,

Aminosulfonyl radical, C₁-C₆Alkylaminosulfonyl, di-C₁-C₆An alkylaminosulfonyl group,

R^bselected from hydrogen, halogen, hydroxy, (C)₁-C₆) Alkoxy, cyano, nitro, amino, hydroxycarbonyl, amino, nitro, cyano, amino,

C₁-C₆alkyl radical, C₁-C₆Alkenyl radical, C₁-C₆Alkynyl, alkynyl,

C₁-C₆alkylsulfonyl radical, C₁-C₆An alkylsulfonylamino group,

10. A compound of formula (Ib) or a pharmaceutically acceptable salt or solvate thereof,

wherein R is^aSelected from halogen, hydroxy, cyano, nitro, amino, hydroxycarbonyl,

C₁-C₆Alkyl radical, C₁-C₆Alkenyl radical, C₁-C₆Alkynyl, alkynyl,

Hydroxy radical C₁-C₆Alkyl radical, C₁-C₆Alkoxy radical C₁-C₆Alkyl, perfluoro C₁-C₆Alkoxy radical,

C₁-C₆alkylsulfonyl radical, C₁-C₆An alkylsulfonylamino group,

An aminosulfonyl group,C₁-C₆Alkylaminosulfonyl, di-C₁-C₆An alkylaminosulfonyl group,

C₁-C₆alkyl radical, C₁-C₆Alkenyl radical, C₁-C₆Alkynyl, alkynyl,

11. A compound of formula (Ic) or a pharmaceutically acceptable salt or solvate thereof:

in the formula R^aAnd R^bIndependently selected from hydrogen, halogen, hydroxyl and (C)₁-C₆) Alkoxy, cyano, nitro, amino, hydroxycarbonyl, amino, nitro, cyano, amino,

C₁-C₆alkyl radical, C₁-C₆Alkenyl radical, C₁-C₆Alkynyl, alkynyl,

C₁-C₆alkylsulfonyl radical, C₁-C₆An alkylsulfonylamino group,

12. A compound of formula (Ia) according to claim 9, which is (2S, 4S) -4- (3-chloro-phenoxy) -pyrrolidine-2-carboxylic acid or a pharmaceutically acceptable salt or solvate thereof.

13. A compound of formula (Ib) according to claim 10, selected from the following:

(2S, 4S) -4- (3-fluoro-benzyl) -pyrrolidine-2-carboxylic acid;

(2S, 4S) -4- (2, 3-difluoro-benzyl) -pyrrolidine-2-carboxylic acid; and

(2S, 4S) -4- (2, 5-difluoro-benzyl) -pyrrolidine-2-carboxylic acid;

or a pharmaceutically acceptable salt or solvate thereof.

14. A compound of formula (Ic) according to claim 11, selected from the following:

(2S, 4S) -4- (3-fluoro-phenoxymethyl) -pyrrolidine-2-carboxylic acid;

(2S, 4S) -4- (3, 6-difluoro-phenoxymethyl) -pyrrolidine-2-carboxylic acid;

(2S, 4S) -4- (2, 3-difluoro-phenoxymethyl) -pyrrolidine-2-carboxylic acid; and

(2S, 4S) -4- (3-methoxy-phenoxymethyl) -pyrrolidine-2-carboxylic acid;

or a pharmaceutically acceptable salt or solvate thereof.