IL134562A - Pyrrolidine carboxylic acid derivatives and methods for their preparation - Google Patents

Abstract

Compounds of the formula where R1 is a C1-C9 straight or branched chain alkyl or C2-C9 straight or branched alkenyl group optinally substituted with C3-C8 cycloalkyl, C3 or C5 cycloalkyl, C5-C7 cycloalkenyl, or Ar1, where aforementioned alkyl, alkenyl, cycloalkyl, or cycloalkenyl groups may optionally have one or more halogen atoms replaced with C1-C4 alkyl, C2-C4 alkenyl or hydroxy, and where Ar1 is selected from the group consisting of 1-naphthyl, 2-naphthyl, 2-indolyl, 3-indolyl, 2-furyl, 3-furyl, 2-thiazolyl, 2-thienyl, 3-thienyl, 2-, 3-, or 4-pyridyl, and phenyl, having one to three substituents which are independently selected from the group consisting of hydrogen, halogen, hydroxyl, nitro, trifluoromethyl, C1-C6 straight or branched alkyl or C2-C6 straight or branched alkenyl, C1-C4 alkoxy or C2-C4 alkenyloxy, phenoxy, benzyloxy, and amino; or pharmaceutically acceptable salts or hydrates thereof. בקשה הורה מס' 121621 לבקשה זו טרם פורסמה. The parent application No. 121621 from which this application has been divided has not yet been published.

Description

134562/3 Pyrrolidine carboxylic acid derivatives and methods for their preparation Guilford Pharmaceuticals Inc.

C. 119913 1 134562/2 BACKGROUND OF TEE ISVEHTION 1. Field of the invention This application is a divisional of Israel Patent Application No. 121621 (hereinafter referred to as the "parent application").

The present application relates, in general, to l-(l,2-dioxo)-2-pyrolidine carboxylic acid derivatives and more specifically to neurotrophic compounds having the above general definition and to their preparation. Such compounds are also used in the preparation of the compounds described in the parent application 2. Description of the grior Art The term irantunophilin refers to a number, of proteins that serve as receptors for the principal " immunosuppressant drugs, cyclosporin A. (CsA) , . FK5QS, and rapamycin. ·' Known classes of iramunophilins- are cyclophilins, and FK5QS binding proteins, such as FK3P. ■ Cyclosporin A binds to cyclophilin while FK5QS and rapamycin bind to FK3?. These immunophilin-drug complexes interface with a variety. of intracellular sicnal transduction systems, especially in the immune ." . ' system and the nervous system.

Immunophilins are known to have peptidyl-prolyl isoraerase (FPIase) cr rotamase enzyme activity. It as been determined that rotamase activity has a role in the cataiyzation of the interconversion of the cis and •trans isomer of immunophilin proteins.

Immunophilins were originally discovered and studied in immune tissue. It was initially postulated by those skilled in the art that inhibition of the immunophilins rotamase activity leads to the inhibition of T-ceil proliferation, thereby causing the immunosuppressive action exhibited by immunosuppressive drugs such as cyclosporin Α F 50S, and rapamycin.

Further study has shown that the inhibition of rotamase activity, in and of itself., is not sufficient for immunosuppressant activity. Schreiber et al., Science, 1990 vol. -250 pp. 55S-559; It has been shown that the immunophilin-drug complexes interac .with ternary protein targets as their mode of action. Schreiber et al., Cell, 1991, vol,-. 66, pp. 807-815. In the case of FXBP-FK506 and FKBP-CsA, ■ the. -drug-immunophilin complexes bind to the enzyme calcineurin, inhibitory T- cell receptor signalling leading to T-cell proliferation. Similarly, the complex of rapamycin and FKSP interacts with. he- RAFT1/FRAP protein and inhibits signalling, from the IL-2 receptor-.

Immunophilins have been found to be present at- high concentrations in the central nervous system.

Immunophilins are enriched 10-50 times more in the central nervous system than in the immune system.

Within neural tissues, immunophiiins a pe to influence neuronal process extension, nitric oxide synthesis, and neurotransmitter release.

It has been found that picomolar concentrations of an immunosuppressant such as FXSQc and rapamycin stimulate neurite out growth in PC12 cells and sensory nervous, namely dorsal root ganglion cells (D Gs) .

Lyons et al., Proc. of Natl. Acad. Sci.f 1994 vol. 91, pp. 3191-3195. In whole animal experiments, F 505 has been shown to stimulate nerve regeneration following facial nerve injury and results in functional recovery in animals with sciatic nerve lesions.

Surprisingly, it has been found that drugs with a high affinity for FKBP are potent rotamase inhibitors causing a neurotrophic effect. Lyons et- al. These findings suggest the Use of immunosuppressants in treating various peripheral neuropathies and enhancing neuronal regrowth in the central nervous system (CNS) . Studies have demonstrated that neurodegenerative disorders such as Alzheimer's disease, Parkinson's disease, and amyotrophic lateral sclerosis (ALS) may occur due to- the loss, br decreased availability, of a neurotrophic substance specific for a particular* . population of neurons affected in the disorder.

Several, neurotrophic factors effecting specific neuronal populations in the central nervous system have been identified. For example, it has been hypothesized that Alzheimer' s disease results from a decrease or loss of nerve growth factor (NG?) . It has thus been proposed to treat Alzheimer's patients with exogenous nerve growth factor or other neurotrophic proteins such as brain derived nerve factor (BDNF) , glial derived nerve factor, ciliary neurotrophic factor, and neurotropin-3 to increase the 'survival of degenerating neuronal populations .

Clinical application of these proteins in various neurological disease states is hampered by difficulties in the delivery and bioavailability of large proteins to nervous system targets. By contrast, immunosuppressant drugs with neurotrophic activity are relatively small and display. excellent bioavailability and specificity. However, when administered chronically, immunosuppressants exhibit a number of potentially serious side effects including nephrotoxicity, such as impairment of glomerular filtration and irreversible interstitial fibrosis ( opp et al., 1991, J. Am. Soc. Nephrol. 1:152); neurological deficits, such as involuntary tremors, or non-specific cerebral angina such as non-localized headaches (De Groen et al., 1987, N. Engl. J. Med. 317:861); and vascular hypertension with complications resulting \ therefrom (Kahan et al . , 1989 N. Engl. J. Med. 321: 1725) .

In order to prevent the side effects associated with use of the immunosuppressant compounds, the present invention provides neurotrophic compounds containing small molecule FK3? rotamase inhibitors for promoting neuronal growth and regeneration in various neuropathological situations where neuronal repair can be facilitated including peripheral nerve damage by physical injury or disease state such as diabetes, physical damage to the central nervous system (spinal cord and brain) brain damage associated with stroke, and for the treatment of neurological disorders relating to neurodegeneration, including Parkinson's disease, Alzheimer's disease, and amyotrophic lateral sclerosis .

SUMMARY OF ΊΈΞ INVENTION The present invention relates to a novel neurotrophic compounds having an affinity for F BP-type immunophilins . Once bound to this protein the neurotrophic compounds are potent inhibitors of the enzyme activity associated with immunophilin proteins and particularly rotamase enzyme activity, thereby stimulating neuronal regeneration and outgrowth. A key feature of the compounds of the present invention is that they do not exert any significant immunosuppressive activity in addition to their neurotrophic activity.

Thus, the invention concerns a compound of the formula: 13 45 6 2 / 3 wher e Ri is selected from the group consisting of a C C9 straight or branched chain alkyl or a C2-C9 straight or branched chain alkenyl group optionally substituted with C3-C8 cycloalkyl; C3 or C5 cycloalkyl; C5-C7 cycloalkenyl or Ari where aforementioned said alkyl, alkenyl, cycloalkyl or cycloalkenyl groups may optionally have one or more hydrogen atoms replaced with Cj-C alkyl, C2-C4 alkenyl, or hydroxy, and where Ai is selected from the group consisting of 1-naphthyl, 2-naphthyl, 2-indolyl, 3-indolyl, 2-furyl, 3-furyI, 2-thiazolyl, 2-thienyl, 3-thienyl, 2-, 3-, or 4-pyridyl, and phenyl, having one to three substitutents which are independently selected from the group consisting of hydrogen, halogen, hydroxyl, nitro, trifluoromethyl, C2-C6 straight or branched alkyl or C2-C6 straight or branched alkenyl, CrC4 alkoxy or C2-C4 alkenyloxy, phenoxy, benzyloxy, and amino; or pharmaceutically acceptable salts or hydrates thereof.

Further, the invention concerns with pharmaceutical compositions comprising a neurotrophically effective amount of the compound of the invention and a pharmaceutically acceptable carrier.

Yet further, the invention relates to a method of synthesizing a chemical compound of the invention, and particularly, (2S)-l-(l,2-dioxo-3,3-dimethylpentyl)-2 -pyrrolidine carboxylic acid, the method comprising the steps of (a) forming a mixture of methyl (2S)-l-(l,2-dioxo-3,3-dimethylpentyl)-2-pyrrolidinecarboxylate, IN lithium hydroxide and methanol; (b) stirring the mixture at 0°C for 30 minutes and at room temperature for about 12 hours; (c) acidifying with IN hydrochloric acid to pH 1; (d) diluting with water; and (e) extracting the organic phase into methylene chloride. -127 ' DESCRIPTION - OF ΞΞ DSAWIHSS Figure 1 is a photomicrograph. of chick dorsal root ganglia treated with various .concentrations of Example 17 as indicated. Figure, 1 shows that Example 17.- of the present invention potently promotes neurite outgrowth in sensory neuronal cultures. Explant cultures isolated form embryonic . day 9 - 10. chick dorsal root ganglia were treated with-various concentrations of 'Example 17 as indicated. . Forty-eight hours later-, the number of neurite with a length greater than one DRG explant was quantitated. The number of neurites . expressed in untreated DRG' s was subtracted form the neurite number of Example 17-treated samples to yield Example 17-dependent specific neurite outgrowth.' δ Micrographs of Example 17 treated ORG' s , as well as quantitative dose-dependent neurite outgrowth elicited by Example 17 are presented.

Figure 2 is a graph showing quantitation of neurite outgrowth in chick dorsal root ganglia treated with various . concentrations of Example 17 as indicated. Figure 2 shows that Example 17 of the present invention potently promotes neurite outgrowth in sensory neuronal cultures. Explant cultures isolated form embryonic day 9 - 10 chick dorsal root ganglia were treated with various concentrations of Example 17 as indicated.

Forty-eight hours later, the number of neurite with a length greater. than one DRG explant was quantitated. The number of neurites expressed in untreated DRG's was subtracted form the neurite number of Example 17-treated samples to yield Example 17-dependent specific neurite outgrowth. Quantitative dose-dependent neurite outgrowth elicited by Example 17. is presented.

Figure 3 is a photomicrograph of rat sciatic nerve sections . Figure 3 shows that Example 1 of the present invention promotes neuronal regeneration following sciatic nerve lesions. Sciatic nerves of 150' g male Sprague-Dawley rats were crushed at the level of the hips. Example 1 (30 mg/kg s.c), Inactive (30 mg/kg s.c.) or intralipid vehicle was administered once daily for the next 21 days. Animals were sacrificed, sciatic nerves removed and nerve 'segments 2 mm distal to the crush, site were sectioned and stained with Holmes silver stain (to assess axon number) and Luxol fast blue (to assess remyeiination) . The micrographs show sciatic nerve, sections of sham operated rats, vehicle-treated lesioned animals, Example 1 and Inactive treated at 63Ox magnification, four animals per group.

Figure 4 is a graph of [3K] -CFT binding per ^g of Striatal Membrane Protein. Figure 4 shows that neuroimmunoph.ilin ligands of the present invention promote recovery of dopamine neurons following MPT? treatment of mice. CD1 mice (25 g) were treated daily with 30 mg/kg MPTP (i.p.) for 5" days. The animals were also treated daily with intralipid vehicle, Example 1 (100 mg/kg s.c.) or Example 17 (40, 20, 10 mg/kg s.c, as indicated) concurrently with the MPT? and continued for an additional 5 days. After eighteen days, the. mice were sacrificed, striata from 5 animals per group were, pooled and processed into a washed membrane preparation. Binding of [3HI -CFT to these striated membrane preparations of various groups was quantitated to determine dopamine transporter levels on viable nerve terminals . Binding in the presence of TO' μΜ unlabelled CFT' provided on estimate of nonspecific binding, which was subtracted from the total binding to quantitative specific [3H] -CFT bound. Binding was normalized to the protein content of the striatal membranes f om ' each experimental group . Coronal and saggital bra..i sections from MPT? a d drug treated animals were stained wit ant tyrosine hydroxylase (TE) Ig to quan i ate striatal, medial forebrain bundle axonal and nigral levels of TK, which is indicative of functional dopaminergic neurons Figure 5 is a bar graph of [3E] -CFT plotted μg of membrane protein.. Figure 5 shows that neuroimiaunoph.ilin ligands of the present, invention promote recovery of dopamine- eurons following MPTP treatment of mice in accordance with the procedure described, in Figure 4.

Figure 6 is a photomicrograph, at.630x magnification, of coronal and saggital brain sections. Figure € shows brain sections from MPTP and drug treated animals stained with an i-tyrosine hydroxylase (TK) Ig to quantitate striatal levels of ΤΉ, which is indicative of functional dopaminergic neurons.

Figure 7 is a photomicrograph, at 5Ox magnification, of coronal and saggital brain sections. Figure 7 shows brain sections from MPTP and drug ' treated animals stained with anti-tyrosine hydroxylase (ΊΉ) Ig to quantitate nigral, levels of TH, which is. ■ indicative of functional dopaminergic neurons .

Figure 8 is a photomicrograph, at 40Ox magnification, of coronal and saggital brain sections Figure 8 shows brain sections from MPTP and drug treated animals stained with anti-tyrosine hydroxylas (TH) Ig to quantitate medial .forebrain bundle axonal levels of TE, which is indicative of functional dopaminergic neurons . ' ' · DS AILSD' DggCSI T O-ff OF THH Ζ2Τ7 ¾ΤΤΟΤΓ The' novel neurotrophic compounds of this inventio are relatively small molecules in relation to other known compounds, which bind to FKBP-type immunophilins, • such as rapamycin, FX5Q6, and cyclosporin.

The neurotrophic compounds of this invention have ' an affinity for the F 506- binding proteins such as F 3P-12. - When the neurotrophic compounds of' the invention are bound to the FX3P, they have been found . to unexpectedly inhibit the prolyl- peptidyl cis-trans isomerase activity,, or rotamase activity of the binding protein and stimulate neurits growth, while not exhibiting an immunosuppressant effect.

More particularly,, this invention relates to ·. neurotrophic compounds represented by . the formula: (I) ' where Rx is a Ci-C9' straight or branched chain alkyl or a C2-C9 straight or branched chain alkenyl group optionally substituted with C3-C8 cycloalkyl ; C3 or C5 cycloalkyl ; C5-C7 cycloalkenyl . or ATI where said alkyl , alkenyl , cycloalkyl or cycloalkenyl groups may optionally have one or more hydrogen atoms substituted with C2-C4 alkenyl, or hydroxy; and where .ATJ. is selected" from- he croup consisting of 1- naphthyl, 2- naphthyl, 2-indolyl, 3-indolyl, 2-furyI, 3-furyI, 2- thiazolyl, 2-thienyl, 3- thienyl, '2-, 3-, 'or 4-pyricyl, or phenyl, avi g one to three substitaents which are independently selected from t e group consisting of hydrogen, halogen, hydroxyl , nitro , trifluoromethyl, Ci-C3 straight or branched alkyl or C2-C3 straight or branched alkenyl , C1-C4 alkoxy or C2-C4 alkenyloxy, phenoxy, benzyloxy, and amino ; 134562/3 13 The parent application relates to the use of neurotrophic compounds which are also relatively small molecules in relation of other known compounds which bind to FKBP-type mimunphilins, such as rapamycin, FK506, and cyclosporin.

The neurotrophic compounds of the parent invention have an affinity for the FK50S- binding proteins such, as FKBP-12. -When the neurotrophic compounds of the invention ar bound to the FKBP, they have been found to unexpectedly, inhibit the prolyl- peptidyl cis-trans isomerase activity, or rotamase activity of the binding protein and stimulate neurits growth, while not exhibiting an imraunosuppressant effect.

More particularly, the parent invention relates to a use of neurotrophic compounds represented by the formula: R1 where ■ ■ Rx is a Ci-Ca straight or branched chain alkyl or a C2-Cg straight or branched chain alkenyl group optionally substituted with C3-C8 cycloalkyl; C3 or C5 cycloalkyl; C5-C7. cycloalkenyl; where said alkyl, alkenyl, cycloalkyl or cycloalkenyl groups may be optionally substituted with C1-C4 alkyl, C2-C4 alkenyl, or hydroxy; or A i, and where .ATJ, is selected' from' ti grots? consisting of 1-napt yl, 2-napt yl, 2-indolyl, 3-indolyl, 2-furyl, 3-furyl, 2- thiazolyl, 2-thienyl, 3- thienyl, 2-, 3-, 'or 4-pyridyl, or phenyl, having one to three s-^bstituents which are independently selected from the group consisting of hydrogen, halogen, hydroxyl, nitro, trifluorometiiyl, Ci-Cg straight or branched alkyl or C2-C6 straight or branched alkenyl, C1-C4 alkoxy or C2-C4 alkenyloxy, phenoxy, benzyloxy, and amino; X is oxygen sulfur, CH2, or two hydrogen atoms , Y is oxygen or NR2, where is hydrogen or Ci~ .

Cs alkyl; and Z is a Cz-Ce straight or branched chain alkyl or alkenyl, wherein the alkyl chain is substituted in one or store positions wish Ar^ as defined ahcvs, C.-C, cycloalkyl, cycicaikyl connected by a -C3 straight or uzibranched alkyl or alkenyl chain, cr Ar where Ar2 is selected from the crous consisting of 2-indoiyi, 3-indciyi, 2= furyl, 3 -furyl, 2- thiazolyl, 2-thienyl, thienyi, 2-, 3-, or 4-pyridyl, or phenyl having one to three substituents which a independently selected front, the group consisting of hydrogen, halo, hydroxyl, nitro, trifiuororaethyl, - straight or branched alkyl or aikenyl, C^C, alkoxy o C2-C4 alkenyloxy, phenoxy, benzyloxy, and mino; may also be the fragment: where R3 -.is selected from the group consisting of ■ straight or branched alkyl C^-C9 optionally substituted with C3-Ca cycloalkyl, cr Ar: as defined above, and unsubstitutsd .¾r,; , 16 134562/2 group consisting of hydrogen, Ci-C6 straight or branched alkyl and C2-C6 straight or branched alkenyl; is selected from the group consisting of phenyl, benzyl, CrC5 straight or branched alkyl or C2-C5 straight or branched alkenyl, and CiC5 straight or branched alkyl or C2-C5 straight or branched alkenyl substituted with phenyl; or pharmaceutically acceptable salts or hydrates thereof, for the preparation of a pharmaceutical composition substantially as described in the specification, for use in a method of promoting neuronal growth and regeneration, treating a neurological disorder, or preventing neurodegeneration, said method comprising administering said pharmaceutical composition in sufficient amounts to promote said neuronal growth and regeneration, treat said neurological disorder, and prevent said neurodegeneration.

II where Rt is a Ci-Cg straight or branched chain alkyl or a C2-C9 straight or branched chain alkenyl group optionally substituted with C3-C8 cycloalkyl, C3 or C5 cycloalkyl, C5-C7 cycloalkenyl, where said alkyl, alkenyl, cycloalkyl or cycloalkenyl groups may be optionally substituted with C1-C4 alkyl, C2-C4 alkenyl, or hydroxy, and where is selec ed from the' g o consisting of' 1-capthyl, 2-sapchyl, 2-indoIyl, 3-iscclyl, 2-furyi, 3-furyl, 2- thiazolyi, 2-thienyl, 3-thienyi, 2-, 3-, or 4-pyridyl, or phenyl, having one to three subsritueats which are independently selected f om the , grous consisting of hydrogen, halo, hydroxy! , nitro, trifluoromethyl , -C3 straight or branched alkyl or C2- e straight or branched. alkenyl, C1-C4 alkoxy or C2-C4 alkenyloxy, phenoxy, benzyloxy, and amino; is a C2-Ce straight or branched chain alkyl or alkenyl, wherein the alkyl chain is substituted in one or more positions with A i as defined above, C3-C8 cycloalkyl, cycloalkyl connected by a Ci-C6 straight or unbranched alkyl or C2-C6 straight or unbranched alkenyl chain, or Ar2 where Ari is selected from the group consisting of 2-indolyl, 3-indolyl, 2- furyl, 3 -fury!, 2- thiazclyl, .· 2-thienyl, 3- . thienyl, 2-, 3-, or 4-pyridyl, or phenyl, having. one tg three su stit ents which are independently selected from the group consisting of hydrogen, halo, hydroxy!, nitro, trifluorcmethyl, -C3 straight or . branched alkyl or C2-C6 straight or branched alkenyl hydrates thereof.

Preferred neurotrophic -glyc-xyl prolyl ester compounds of the parent application have the formula: where Ri is a C1-C5 straight or branched chain alkyl or C2-C5 straight or branched alkenyl group optionally substituted with C3 to Z cycloalkyl, or Arj., where Ari is selected from the group consisting of 2- furyl, 2-thienyl, or phenyl; is selected from the group consisting of oxygen and sulfur; Y is oxygen; and z is a straight or branched chain alkyl or alkenyl, wherein the alkyl chain is substituted in one or more positions with Ar, as defined above, C3-C3 cycloalkyl, Ar, where 2-, 3-, or -pyridyl, or phenyl, having ons to three substituents which are indeDendently selected from the group consisting of hydrogen and C,-C4 alkoxy.

The compounds of this invention exist as stereoisomeric forms, either enantiomers or diastereoisomers . The stereochemistry at position 1 (Formula 1) is R or S, with 3 preferred. Included within the scope of the invention are the enantiomers, the racemic form, and diastereoisomeric mixtures.

Enantiomers as well as diastereoisomers can be separated by methods known to those skilled in the art.

It is known that immunophilins such as F BP preferentially recognize peptide substrates containing Xaa-Pro-Yaa motifs, where Xaa and Yaa are lipophilic amino acid residues. Schreiber et al. 1990 J". Org*.

Chem. 55, 4984-4986; Harrison and Stein, 1990 Biochemis ry, 29, 3813-38Ί6. Thus modified prolyl peptidomimetic compounds bearing lipophilic substituents should bind with high affinity to the hydrophobic core of the FKBP active site and inhibit its rotamase activity.

Preferred compounds of the present invention include RL groups which are not stereochemical^ bulky in relation to the known shape and size of the hydrophobic core of the FK3? active site. Thus, very-large and/or highly substituted : groups would bind with less affinity to the FIG? active sits.

Preferred compounds of the invention include: 3 -phenyl-1-propyl (2S) -I- (3, 3 -dimethyl-1,2-dioxopentyl) -.2-pyrrolldinecarboxylate, 3 -phenyl-1-prop-2- (Ξ) -enyl (2S) -1- (3 , 3 -dimethyl-1, 2-dioxopentyl) -2-pyrrolidinecarboxylate, 3- (3,4, 5-trimethoxyphenyl) -1-propyl (2S)-l-(3,3-dimethyl-1, 2-dioxopentyl) -2-pyrrolidinecarboxylate, 3- (3 ,4, 5-trimethoxyphenyl) -l-prop-2- (E) -enyl (2S) 1- ( , 3 -dimethyl-1, 2-dioxopentyl) -2-pyrrolidine carboxylate, 3- (4, 5-methylenedioxyphenyl) -l-propyl (2S)-l-(3,3 dimethyl-1, 2-dioxopentyl) -2-pyrrolidinecarboxylate, 3- (4, 5-methylenedioxyphenyl) -l-prop-2- (E) -enyl (2S) -1- (3, 3-dime hyl-1, 2-dioxopentyl) -2-pyrrolidinecarboxylate, 3 -cyclohexyl-l-propyl (2S) -1- ( , 3 -dimethyl-1, 2-dioxopentyl) -2-pyrrolldinecarboxylate, 3-cyclohexyl-l-prop-2- (E) -enyl (2S) -1- (3,3-dimethyl-1, 2-dioxopentyl) -2-pyrrolidinecarboxylate, ■ (1R) -l, 3-diphenyl-l-propyl (2S) -1- (3 , 3 -dimethyl-1, 2-dioxopentyl) -2-pyrrolidinecarboxylate, 3 -phenyl-l-propyl (2S) -1- (1, 2-dioxo-2- [2-furanyl] )■ethyl-2-pyrrolidinecarboxylate, 3 -phenyl-l-propyl (2S) -1- (1, 2-dioxo-2- [2-thienyl] ) ethyl-2-pyrrolidinecarboxylate, 3 -phenyl-l-propyl (2S) -1- (1 , 2-dioxo-2- [2-thiazolyl] ) ethyl-2-pyrrolldinecarboxylate , 3 -phenyl-1-propyl (2S) -I- (1, 2-dioxo-2- ph.er.yl ) ethyl -2 -pyrrolidinecarboxylate , 3- (2, 5-dimethoxyphenyl) -1-propyl (2S)-l-(3,3- dimethyl-1, 2 -dioxopentyl) -2-pyrrolidinecarboxylate, 3- (2 , 5-dimethoxyphenyl) -1-prop-2- (E) -enyl (2S) - (3 , 3 -dimethyl-l# 2 -dioxopentyl) -2-pyrrolidinecarboxylate, 2- (3,4, S-trimethoxyphenyl) -1-ethyl (2S)-l-(3,3- dimethyl-1, 2.-dioxopentyl) -2-pyrrolidinecarboxylate, 3- (3-Pyridyl) -1-propyl (2S) -1- (3 , 3-dimethyl-l, 2- dioxope t'yl) -2 -pyrro1idinecarboxy1ate , 3- (2-Pyridyl) -1-propyl (2S) -1- (3 , 3-dimethyl-l, 2- dioxopentyl) -2-pyrrolidinecarboxylate, 3- (4-Pyridyl) -1-propyl (2S) -1- (3 , 3 -dimethyl-1, 2- dioxopentyl) -2-pyrrolidinecarboxylate, 3 -phenyl-1-propyl (2S) -1- (2-cyclohexyl-l, 2- . dioxoethyl) -2 -pyrrolidinecarboxylate , 3-phenyl-1-propyl (2S) -1- (2- tert-butyl-1, 2- dioxoethyl) -2-pyrrolidinecarboxylate,. 3 -phenyl-1-propyl (2S) -1- (2-cyclohexylethyl-l, 2- ' dioxoethyl) -2-pyrrolidinecarboxylate, 3- (3-pyridyl) -1-propyl (2S) -1- (2-cyclohexylethyl 1, 2-dioxoethyl) -2-pyrrolidinecarboxylate, 3- (3-pyridyl) -1-propyl (2S) -1- (2- tert-butyl-1, 2- dioxoethyl) -2-pyrrolidinecarboxylate, 3 , -diphenyl-1-propyl (2S) -1- (3 , 3-dimethyl-l, 2 - dioxopentyl) -2 -pyrrolidinecarboxylate, 3- (3-pyridyl) -1-propyl (2S) -1- (2-cyclohexyl-l, 2- cioxoethyl) -2-pyrrolidinecarboxylate , 3- (3-Pyridyl) -1-propyl (2S) -N- ( [2-t ienyl] glyoxyl) pyrrolidinecarboxyla e, 3, 3-Diphenyl-l-propyl (2S) -1- (3 , 3 -dimethyl-1, 2- dioxobutyl) -2-pyrrolidinecarboxylate, 3 , 3 -Diphenyl-l-propyl (2S) -1-cyclohexylglyoxyi- 2 -pyrrolidinecarboxylate , 3 , 3-Diphenyl-l-propyl (2S) -1- (2-thisnyl) glyoxyl- 2 -pyrrolidinecarboxylate .

Particularly preferred neurotrophic N-glyoxyl prolyl ester compounds are selected from the group consisting of: 3- (2, 5-dimethoxyphenyl)-I-propyl (2S)-l-(3,3- dimethyl-1, 2-dioxopentyl) -2 -pyrrolidinecarboxylate, 3- (2, 5-dimethoxyphenyl) -l-prop-2- (Ξ) -enyl (2S)-1- (3 , 3 -dimethyl-1, 2-dioxopentyl) -2 -pyrrolidinecarboxylate, 2- (3,4,5 -trimethoxyphenyl) -1-ethyl (2S)-l-(3,3-' dimethyl-1, 2-dioxopentyl) -2-pyrrolidinecarboxylate, 3- (3-Pyridyl) -1-propyl (2S) -1- (3 , 3 -dimethyl-1, 2- dioxopentyl ) -2 -pyrrolidinecarboxyla e , 3- (2-Pyridyl) -1-propyl (2S) -1- (3 , -dimethyl-1, 2- dioxopentyl) -2-pyrrolidinecarboxylate , 3- (4-Pyridyl) -l-propyl (2S) -1- (3 , 3 -dimethyl-1, 2- dioxopentyl) -2-pyrrolidinecarboxylate, 3 -phenyl -1 -propyl _(2S) -1- (2- Cert-butyl-1, 2- dioxoe hyl ) -2 -pyrrolidinecarboxylate , 3 -phenyl -1 -propyl (2S) -1- (2-cyclohexylethyl-l, 2- dioxoethyl) -2-pyrrolidinecarboxylate, 3- (3-pyridyl) -l-propyl (2S) -1- (2-cyclohexylethyl-1, 2-dioxoethyl) -2 -pyrrolidinecarboxylate, 3-,(3-pyridyl) -l-propyl (2S) -1- (2- Cert-butyl-l, 2-dioxoethyl) -2-pyrrolidinecarboxylate, 3, 3 -diphenyl-l-propyl (2S) -1- (3 , 3 -dimethyl-1, 2 -dioxopentyl ) - 2 -pyrrolidinecarboxylate , 3- (3-pyridyl) -l-propyl (2S) -1- (2-cyclohexyl-l, 2-dioxoethyl) -2-pyrrolidinecarboxylate, 3- (3-Pyridyl) -i-propyl (2S) -N- ( [2-thienyl] glyoxyl) pyrrolidinecarboxylate, 3, 3 -Diphenyl-l-propyl (2S) -1- (3 , 3-dimethyl-l,2-dioxobutyl) -2-pyrrolidinecarboxylate, 3 , 3 -Diphenyl -l-propyl (2S) -1-eyelohexylglyoxyl -2-pyrrolidinecarboxylate, and 3 , 3 -Diphenyl-l-propyl (2S) -1- (2-thienyl) glyoxyl-2 -pyrrolidinecarboxylate .

The compounds of the present invention can be used in the form of salts derived from inorganic or organic acids and bases . Included among such acid salts are the following: acetate, adipate, alginate, aspartate, benzoate, benzenesulfonate, bisulfate butyrate, citrate, camphorate, camphorsulfonate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, fumarate, glucoheptanoate, glycerophosphate, hemissulfate heptanoate, hexanoate, hydrochloride, hydrobromide , hydroiodide, 2- hycLrcxyethanesulfonate, lactate, raaleate, methanesulfonate, 2-napht-jaalensulfonate, nicotinate, oxalate, pamoate, pectinate, propionate, succinate, tartrate, . thiocyanate, tosylate and undecanoate. Base salts include ammonium salts, alkali metal salts such as sodium and potassium salts, alkaline earth metal salts such as calcium and magnesium salts, salt with organic bases such as dicyclohexylamine salts, N-methyl-D-glucamine, and salts with amino acids such as arginine, lysine, and so forth. Also, the basic nitrogen-containing groups can be quartemized with such agents as lower alkyl halides, such as methyl, ethyl , propyl, and butyl chlorid , bromides and ' iodides; dialkyl sulfates like dimethyl, diethyl, dibutyl and diamyl sulfates, long chain halides such as decyl, lauryl, myristyl and stearyl chlorides, bromides and iodides, aralkyl halides like benzyl and phenethyl bromides and others. Water or oil-soluble or dispersible products are thereby obtained.

The neurotrophic compounds"of this invention can be periodically administered to a patient undergoing treatment for neurological disorders or for other reasons in which it is desirable to stimulate neuronal regeneration and growth, such as in various peripheral neuropathic and neurological disorders relating to neurodegenera ion. The compounds of this invention ear-also be administered to mammals other than humans for treatment of various mammalian neurological disorders.

The novel compounds of the present invention are potent inhibitors of rotamase activity and possess an excellent degree of neurotrophic activity. This activity is useful in the stimulation of damaged neurons, the promotion of neuronal regeneration, .the prevention of neurodegeneration, and in the treatment of several neurological disorders known to be associated with neuronal degeneration and peripheral neuropathies. The neurological disorders that may be treated include but are not limited to: trigeminal neuralgia-, glossopharyngeal neuralgia, Bell's Palsy, myasthenia gravis, muscular dystrophy, amyotrophic lateral sclerosis, progressive muscular atrophy, progressive bulbar inherited muscular atrophy, herniated, ruptured or prolapsed invertabrae disk syndromes, cervical spondylosis, plexus disorders, thoracic outlet destruction syndromes, peripheral neuropathic such as those caused by lead, 'dapsone, ticks, prophyria, or Gullain-Barre syndrome, Alzheimer's disease, and Parkinson's disease.

For these purposes the compounds of the present invention may be administered orally, parenterally, by inhalation spray, topically, rectally, nasally, -buccally, vaginally or via an implanted reservoir m dosage formulations containing conventional non-toxic pharmaceutically-acceptable carriers, adjuvants and vehicles . The term parenteral as used herein includes subcutaneous, intravenous, intramuscular, intrapsritonsally, intrat ecally, intraventricularly, intrasteraal and intracranial injection or infusion techniques.

To be effective therapeutically as central nervous system targets the immuno hilin-drug complex should readily penetrate the blood-brain barrier when peripherally administered. Compounds of this invention which cannot penetrate the blood-brain barrier can be effectively administered by an intraventricular route . The pharmaceutical compositions may be in the form of a sterile injectable preparation, for example as a sterile injectable aqueous or oleaginous suspension. This suspension may be formulated according to techniques know in the art using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally- . acceptable diluent or solvent, for example as a solution in 1, 3-butanediol . Among the acceptable vehicles nd^ solvents that may be employed are water, Ringer' s solution and isotonic sodium chloride solution. In addition', sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose any bland fixed oil may be employed including synthetic mono- or diglycerides . Fatty acids such as oleic acid and its glyceride derivatives find use in the preparation of injectables, olive oil or castor oil, especially in their polyoxyethylated versions. These oil solutions or suspensions may also contain a long-chain alcohol diluent or dispersant.

The compounds may be administered orally in the form of capsules or tablets, for example, or as an aqueous suspension or solution. In the case of tablets for oral use, carriers which are commonly used includ lactose and corn starch. Lubricating agents, such as magnesium stearate, are also typically added. For oral administration in a capsule form, useful diluents include lactose and dried com starch. When aqueous suspensions are required for oral use, the active ingredient is combined with emulsifying- and suspending agents. If desired, certain sweetening and/or flavoring and/or coloring agents may be added.

The compounds of this invention may also be administered in the form of suppositories for rectal administration of the drug. These compositions can be . prepared by mixing the drug with a. suitable non-irritating excipient which is solid at room temperature but liquid at rectal temperature and therefore will melt in the rectum to release the drug. Such materials include cocoa butter, beeswax and polyethylene glycols. The compounds of this invention may also be administered optically, especially when the conditions addressed for treatment involve areas or organs readily accessible by topical application, including neurological disorders of the eye, the skin, or the lower intestinal tract. Suitable topical formulations are readily prepared for each of these areas .

For ophthalmic use, the compounds can be formulated as micronized .suspensions in isotonic, pK adjusted sterile saline, or, preferably, as solutions is isotonic, pH adjusted sterile saline, either with or without a preservative such as benzylalkonium chloride . Alternatively for the ophthalmic uses the compounds may be formulated in an ointment such as petrolatum.

For application topically to the skin, the compounds- can be formulated- in a suitable ointment containing the compound suspended or dissolved in, for example, mixture with one or more of the following: mineral oil, liquid petrolatum, white petrolatum, propylene glycol, polyoxyethylene polyoxypropylene compound, emulsifying wax and water. Alternatively, the compounds can be formulated in a suitable lotion or cream containing the active compound suspended or dissolved in, for example, a mixture of one or more of the following: mineral oil, sorbitan monostearate, polysorbate SO, cetyl esters wax, cetearyl alcohol, 2-octyldodecanol , benzyl alcohol and water.

Topical application for the lower intestinal" tract an be effected in a rectal suppository formulation (see above) or in a suitable enema formulation.

Dosage levels on the order of about .Img to about 10,000 mg. of the active ingredient compound are useful in the treatment of the above conditions, with preferred levels cf about O.Xmg to about 1,000 tng. Th. amount cf active ingredient that may be combined with the carrier materials to produce a single dosage form will vary depending upon the host treated and the particular mode of administration.

It is understood, however, that a specific dose level for any particular patient will depend upon a variety of factors including the activity of the specific compound employed, the age, body weight, general health, sex, diet, time of administration, rat of excretion, drug combination, and the severity of th particular disease being treated and form of administration.

The compounds can be administered with othe neurotrophic agents such as neurotrophic growth factor (NGF) , glial derived growth factor, brain derived growth factor, ciliary neurotrophic factor, and neurotropin-3. The dosage level of other neurotrophic drugs will depend upon the factors previously stated . and the neurotrophic effectiveness of the drug combination.

A preferred compound according to the invention is (2S)-l-(l,2-dioxo-3,3-dimethyl pentyl)-2-pyrrolidine carboxylic acid. Thus, the invention also concerns with a method of synthesizing this compound, which method comprises the steps of (a) forming a mixture of methyl (2S) -(l,2-dioxo-3,3-dime lpentyl)-2-pyrrolidinecarboxylate, IN lithium hydroxide and methanol; (b) stirring the mixture at 0°C for 30 minutes and at room temperature for about 12 hours; (c) acidifying with IN hydrochloric acid to pH 1; (d) diluting with water; and (e) extracting the organic phase into methylene chloride. i Test Procedure Inhibition of the pe tidyl-prolyl isomerase" · (rotamase) activity of the inventive compounds can .be evaluated by known methods described in the literature (Harding, M.W. et al. Nature 341: 758-760 (1989); Holt et al. J. Am. Che . Sac. 115: 9923-9938). These values are obtained as aonarent t' s and are presented in Table I. The cis- trans isomerization of an alanine- proline bond in a model substrate, H-succinyl-Ala-Ala-■ Pro-Phe-p-nitroanilide,. is monitored . spectrophotometrically in a chymotrypsin-coupled assay, which releases para-nitroanilide from the trans form of the substrate. The inhibition of this reaction caused by the addition of different concentrations of inhibitor is determined, and the data is analyzed as a change in first-order rate constant as a function of inhibitor concentration to yield; the apparent values · ■ In a plastic cuvette are added 950 mL of ice cold assay buffer (25 mM HEPES, pE 7.8, 100 mM NaCl) , 10 mL of F B? (2.5 mM in 10 mM Tris-Cl pH 7.5, 100 mM NaCl, 1 mM dithiothreitol) , 25 mL of chymotrypsin (50 mg/ml in l mM HCl) and 10 mL of test compound at various concentrations in dimethyl, sulfoxide. The reaction is _ initiated by the addition of 5 mL of substrate (succinyl-Ala-Phe-Pro-Phe-para-nitroanilide, 5 mg/mL in 2.35 mM LiCl in trifluoroethanol) .

The absorbance at .390 nm versus time is monitored for 90 sec using a spectrophotometer and the rate constants are determined from the absorbance versus time data files.

The data for these experiments is presented' in Table I.

No. R X -dimethylpropyl 3-phenylpropyl 42 3 -phenyl-prop-2- 12S (£) -enyl 3- (3,4* 5-tri- . methoxypheayl) propyl 200 4 3- (3 , , 5-trimethoxy- phenyl) -prop-2- (E) -enyl S3 3 - (4, Ξ-me hylenedioxy) phenylpropyl 170 3- (4, 5-mechylenedicxy) phenylprop-2- (Ξ) -enyl ISO 7 3-cyelchexylpropyl 200 a 3-cyciohexylprop-2- (3} - enyl 600 (1R) -1,3-diphenyl-l- propyl 52 2-furanyl 3-phenylpropyl 4000 11 2-chiftTryl S2 12 2-chiazclyl 100 13 ph.ea.yl .1370 14 1, 1-dimethylpropyl 3- (2,5-dimethcxy) phenylpropyl 250 IS 3- (2, 5-dimethcxy) phenylprop-2- (Ξ) -enyl 450 IS 2- (3 , 4*; 5-erimechoxy phenyl) ethyl 120 17 3- (3-pyridyl) propyl 5 ia 3- (2-pyridyl) propyl 135 13 3- (4-pyridyl) propyl 23 20 cycichexyl 3-;phenylpropyl 82 21 cert-butyl " 95 22 cyclohexylethyl " 1025 23 cyclohexylethyl · 3 - {3-pyridyl) propyl 1400 24 cert-butyl 3 -{3-pyridyl) propyl 3 1, l-dimetiiylpropyl . 3 , -diphenylpropyl 5 25 cyclohexyl 3 - {3-pyridyl) propyl 27 2-thianvi 3 - (3-pyridyl) propyl 1000 28 tert-buttyl 3 , 3-diphenylpropyl 5 23 cyclohexyl. " 20 2-thianyl ■150 In mammalian cells, F 3F-12 complexes with, the inositol triphosphate receptor (I?3R) and the ryanodine receptor (RyR) . It is believed that the neurotrophic compounds of this invention disassociates F 3P-I2 from these complexes causing the calcium channel to become "leaky" (Cameron et al . , 1995) . Calcium fluxes are involved in neurite extensions so that the IP3R receptor and the ryanodine receptor might be involved in the neurotrophic effects of drugs. Since the drugs bind to the same site as F 3P-12 as the IP3R receptor, one could assume that the drugs displace the channels from F BP-12.

Chick Dorsal Root Ganglion Cultures and Neurite Outgrowth r Dorsal'- root ganglia were dissected from chick embryos of ten day gestation. Whole ganglion explants were cultured on thin layer Matrigel-coated 12 well plates with Liebovitz LIS plus high glucose media supplemented with 2mM glutamine and 10% fetal calf serum, and also containing 10 μΜ cytosine /3-D arabinofuranoside (Ara C) at 37°C in ah environment containing 5% C02. Twenty-four hours later, the DRGs were treated with various concentrations of nerve* growth factor, immunophilin ligands or combinations of NFG plus drugs. Forty-eight hours after drug treatment, the ganglia were visualised under phase contrast or Hoffman Modulation contrast with a Zeiss Axiovert inverted microscope. Photomicrographs of the explan s were made, and neurite outgrowth was quantitated. Neurites longer than the ORG diameter were counted as positive, with total number of neurites quantitated per each experimental condition. Three to four ORGs are cultured per well, . nd each treatment was performed in duplicate..

The data for these experiments are presented in Table II. Representative photomicrographs for Exanrole 17 are shown in Figure 1; a dose response curve for this Example is given in Figure 2.

TABLE II Neurite Outgrowth, in Chick .DUG Example No . E EDD5500,, neurite outgrowth, nM 1 53 2 105 3 149 4 190 10 S 75 0.46 11 0.015 14 2 0.8 IS 0.015 17 0.05 18 3 19 6 - 20 0.13 21 0.025 22 0.66 23 1100 24 0.014 0.50 26 2 27 500 23 0.50 29 10 100 Sciatic Nerve Axotomy Six-week old male S^raoue-Dawlev rats were anesthetized, and the sciatic nerve exposed and crushed, at the level of the hip, by forceps. Test compounds or vehicle were administered subcutaneousiy just prior to the lesion and daily for the following 18 days . Sections of the sciatic nerve were stained with Holmes silver stain .to quantify the number of axons, and Luxol fast blue to quantify the level of myelination. Eighteen days after lesion, there was a significant decrease in the number of axons (50% decrease as compared to non-lesioned control) and degree of myelination (90% decrease as compared to non- lesioned 'control) in animal treated with vehicle.

Administration of Example 1 (30 mg/kg s.c), just prior to the lesion and daily for 18 days following the lesion, resulted in significant regeneration of both axon number (5% decrease as compared to non-lesioned control) and the degree of myelination (50% decrease as compared to control) as compared to vehicle treated animals. The significant efficacy of Example 1 is . consistent with its potent activity in inhibiting rotamase activity and stimulating neurite outgrowth in chick DRGs. These results are shown in Figure 3. "Sham" .denotes control animals that received, vehicle but" ere ■ not lesioned "Vehicle" denotes animals that were lesioned and received only vehicle (i.e., no drug).

Example l showed a striking similarity to the sham ■ treated animals, demonstrating the powerful neurorecenerative effects of these compounds i -vivo.

Inactive is a compound that is inactive as an FK3P12 inhibitor. Animals treated with this compound resembled the vehicle-treated lesioned animals, consistent with the neuroregenerative results observed with Example 1 being directly caused by its inhibition of FK3P12.

Quantitation for these data are shown in Table III.

Table III Treatment Axon Number Myelin Level ( Control) Sham 100 100 Lesion: + . Vehicle (s.c.) 50 10 + ExamDle 1 ■ 100 ' 50 (30 mg? kg s . c . ) + Inactive 25 25 (30 mg/kg s.c.) MPTP Model of Parkinson's Disease in Mice MPT? lesioning of dopaminergic neurons in mice was used as an animal model of Parkinso '.s Disease. Four week old male CD1 white mice were dosed i.p. with 30 mg/kg of MPTP for 5 days. Example 17(10-40 mg/kg), or vehicle, were administered s.c. along with the MPTP for 5 days, as well as for 'an additional 5 days following cessation of MPTP treatment. At 18 days following MPTP treatment, the animals were sacrificed and the striata were dissected and homogenized.. Binding of [3E]CFT, a. radioligand for the dopamine transporter, to the stiatal membranes was done to quantitate the level of the dopamine transporter (DAT) following lesion and drug treatment . Immunostaining was performed on saggital and coronal brain sections using anti-tyrosine hydoxyiase Ig to quantitate survival and recovery of dopaminergic neurons. In- animals treated with MPTP and vehicle/ a substantial loss of functional dopaminergic terminals was observed as compared to non-lesioned animals. Lesioned animals receiving Example 17 showed a nearly quantitative recovery of TK-stained dopaminergic neurons .

Figures 4 and 5 show the quantitation in DAT levels, whereas figures 6-8 are photomicrographs showing the regenerative effects of Example 17 in this-model.' Figure 4 demonstrates the significant recovery in functional dopaminergic terminals, as assayed by [3.H] -CFT binding, relative to animals receiving MPTP but . not the Guilford compounds . Figure 5 gives this data in bar graph form. It is shown tha animals ■ receiving 40 mg/kg of Example 17 in addition to MPTP manifested a greater than 90% recovery of [3H] -CFT binding. As shown in Figures 6-8, immunostaining 'for tyrosine hydroxylase (a marker of viable dopaminergic neurons) in the striatum, the nigra, and the medial forebrain bundle, shows a clear and marked recovery of functional neurons in animals that received Example 17, as compared to. animals that received lesioning agent but no drug (MPT?/Vehicle) .

The following examples are illustrative of preferred embodiments of the invention and are not to be construed as limiting the invention thereto. All preferred embodiments of the invention and are not to be construed as limiting the invention thereto. All polymer molecular weights are mean average molecular weights,. All percentages are based on the percent by weight of the final delivery system or formulation prepared unless otherwise indicated and all totals equal 100% by weight.

EXAMPLES The inventive compounds may be prepared by a variety of synthetic sequences that utilize established chemical .transformations. The general pathway to the present compounds is described in Scheme .1. N-glyoxylproline derivatives may be prepared by reacting L-proline methyl ester' with methyl oxalyl chloride as shown in Scheme I. The resulting oxamates may be reacted with a variety of carbon nucleophiles to obtain intermediates compounds. These intermediates are then reacted with a variety of alcohols , . amides, or protected amino acid residues to obtain the propyl esters and amides of the invention.

Scheme I SSAMPLS 1 Synthesis of 3 -phenyl-1-propyl (2S) -l- (3 , 3-dimeth l-l , 2-dioxopentyl) -2-pyrrolidinecarboxylate (Example 1).

Synthesis of methyl (2S) -1- (1, 2-dioxo-2-methoxyethyl) -2-pyrrolidinecarboxylate..

A solution of L-proline methyl ester hydrochloride (3.08 g; 18.60 mmol) in dry methylene chloride was cooled to 0°C and treated with triethylamine (3.92 g; 38.74 mmol; 2.1 eq) . After stirring .the formed slurry under a. itrogen, atmosphere for 15 min, a solution of methyl oxalyl chloride (3.20 g; 25.12 mmol) in methylene chloride (.45 mL) was added dropwise. The resulting mixture was stirred at 0°C for 1.5 hr. After filtering to remove solids, the organic phase was washed with water, dried over MgS04 and concentrated.

The crude residue was purified on a silica gel column, eluting with 50% ethyl acetate in hexane, to obtain 3.52 g (83%) of the product as a reddish oil. Mixture of cis-trans amide rotamers; data for trans rotamer given. ¾ NMR (CDC13).: d .1.93 (dm, 2E) ; 2.17 (m, 2E) ; 3.62 (m, 2E) ; 3.71 (s, 3K) ; 3.79, 3.84 ( S, 3K total); 4.85 (dd, IK, J = 8.4, .3.3) .

Synthesis of methyl (2S) -1- (1, 2-dioxo-3 , 3- dimethylpentyl) -2-pyrrolidinecarboxylate.

A solution of methyl (2Sj -1- (l, 2-dioxo-2- methoxyethyl) -2-pyrrolidinecarboxylate (2.35 g; 10.90 mmol) in 30 mL of tetrahydrofuran (THF) was cooled to -78°C and treated with 14.2 mL of a 1·.0 M solution of 1, 1-dimethylpropylmagnesium chloride in TKF. After stirring the resulting . homogeneous mixture at -78°C for three hours, the mixture was poured into saturated ammonium chloride (100 -mL) and extracted into ethyl acetate. The organic phase was washed with water, dried, and concentrated, and the crude material obtained upon removal of the solvent was purified on a silica gel column, eluting with 25% ethyl acetate in hexane, to obtain 2.10 g (75%) of the oxamate as a colorless oil. ¾ MR (CDC13) : d Ό..88 (t, 3HO ; 1".22, 1.26 (s, 3K each); 1.75 (dm, 2H) ; 1.87-2.10 (m, 3H) ; 2.23 (m, IK);- 3.54 (m, 2H) ; 3.7S (s, 3H) ; 4.52 (dm, 1H, ' J = 8.4, 3.4).

Synthesis of (2S) -i- (1, 2-dioxo-3 , 3 - dimethyipentyl) -2-pyrrolidinecarboxylic acid.

A mixture of methyl (2S) -1- (1, 2-dioxo-3 ,3-dimethyipentyl) -2-pyrrolidir.ecarboxylate (2.10 g; 8.23 mmol) , 1 N LiOK (15 mL) , and methanol (50 tnL) was ■ stirred' at 0°C for 30 min and at room temperature overnight. The mixture was acidified to pH l with 1 N HC1, diluted with water, and extracted into 100 mL of methylene chloride . ' The organic extract was washed with brine, and concentrated to deliver 1.73 g (87%) of snow-white solid which did not require further purification. ¾ MR (CDC13) : d 0.87 (t, 3K) ; 1.22, 1.25 (s, 3H each); 1:71 (dm, 2H) ; 2.02 (m, 2H) ; 2.17 (m, 1H) ; 2.25 (m, 1H) ; 3.53 (dd, 2K, J = 10.4,· 7.3); 4.55 (dd, 1H, J = 8.6, 4.1).

Synthesis of 3 -phenyl-1-propyl (2S)-l-(3,3-dimethyl-1,2-dioxopentyl) -2-pyrrolidinecarboxylate (Example 1). A mixture of (2S) -1- (1, 2-dioxo-3 , 3-dimethylpentyl) -2-pyrrolidine-carboxylic acid (600 mg; 2.49 mrnol), 3 -phenyl-1-propanol (508 mg; 3.73 mrnol), dicyclohexylcarbodiimide- (822 mg; 3.98 mrnol), camphorsulphonic acid (19-0 mg; 0.8 mmol) and 4-dimethylaminopyridine , (100 mg; 0.8 mmol) in methylene chloride (20 mL) was stirred overnight under a nitrogen atmosphere. The reaction mixture was filtered through Celite to remove solids and concentrated in vacuo, and the crude material was purified on a flash column (25% ethyl acetate in hexane) to obtain 720 mg (80%) of Example 1 as a colorless oil. ¾ NMR (CDC13) : d 0.84 (t, 3K) ; 1-19 (3, 3H) ; 1.23 (s, 3H) ; 1-70 (dm, 2H) ; 1.98 (m, 5H) ; 2.22 (ra, IE); 2.54 (m, 2Ξ);. 3.47 (m, 2H) 4.14 Cm, 2H) ; 4.51 (d, 1H) ; 7. IS (m, 3Ξ) ; 7.25 (m, 2H) The method of Example 1 was utilized to preaare the following illustrative examples : Example 2: 3 -phenyl-1-prop-2- .(E) -enyl (2S) -1- (3 , 3-dimethyl-1, 2-dioxopentyl) -2-pyrrolidinecarboxylate, 80%, ¾ S (3S0 Mhz, CDC13) : d 0.86 (t, 3E) ; 1.21 (s, 3H) ; 1.25 (s, 3H) ; 1.54-2.10 (m, 5H) ; 2.10-2.37 (m, IK); 3.52-3.55 (m, 2H) ; 4.55 (dd, IE, J = 3.8, 8.9); 4.78-4.83 (m, 2E) ; 6.27 (m, IE); 6.67 (dd, IE, J = 15.9); 7.13-7:50 (m, 5H) .- Example 3: 3 - (3 , , 5-trimethoxyphenyl) -1-propyl (2S) -1- ( , 3-dimethyl-l, 2-dioxopentyl) -2-pyrrolidinecarboxylate, 51%, ¾ ffi (C CLj) : d 0.84 (t, 3E) ; 1. IS (s, 3E) ; 1.24 (s, 3E) ; 1,71 (dm, 2E) ; 1.98 (m, 5E) ; 2.24 (m, IE); 2.53 (m, 2E) .; 3.51 (t, 2E) ; 3.79 (s, 3H) ; 3.83 (s, 3E) ; 4.14 (m, 2E) ; 4.52 (m, IS); 6.36 (s, 2H).

Example 4 : 3- (3,4, 5-trimethoxyphenyl) -l-prop-2- (E) -enyl (2S) -1- (3 , 3-dimethyl-l, 2-dioxopentyl) -2 -pyrrolidine carboxylate, 56%, ¾ TO (CDC13) : d 0.85 (t, 3E) ; 1.22 (S, 3E) ; 1.25 (s, 3E) ; 1.50-2.11 (m, 5H) ; 2..11-2.40 (m, 1H) ; 3.55 (m, 2H) ; 3.85 (s, 3H) ; 3.88 (s, 6H) ; 4.56 .(dd, IE); 4.81 (m, 2E) ; 6.22 (m, IE); 6.58 (d, IE, J = 16) ; 6.53 (s, 2E) .

Example 5: 3 - ( , 5-metnyisnedioxyphenyl) -1-propyl (2S)-1- (3 , 3 -dimethyl -1, 2-dioxopentyl) -2 -pyrrolidine- ■ Example 10: 3 -phenyl-1-propyl (2S) -1- (l, 2-dioxo-2- [2-furanyl] ) ethyl-2-pyrrolidi2iecarboxylate/ 99%, ¾ NMR (300 MHz , CDC13) : d 1.65-2.41 (m, 6E) ; 2.72 (t, 2E, J = 7.5); 3.75 (m, 2E).; 4.21 (m, 2E) ; 4. SI (m, 1H) 7 6.58 (m, IE); 7.16-7.29 (m, 5H) ; 7.73 (m, 2E) .

Example 11: 3 -phenyl-1-propyl (2S) -1- (l, 2-dioxo-2- [2-thienyl] ) ethyl-2-pyrrolidinecarboxylate, 8.1%, 1H NMR (300 MHz , CDCI3) : d 1.88-2.41 (m, SE) ; 2.72 (dm, 2E) ; 3.72 (m, 2E) ; 4.05 (m, IE); 4.22 (m, IE); 4.64 (m, IS); 7.13-7.29 (m, 6H) ; 7.75 (dm, IE); 8.05 (m, IS) .

Example 13 : -phenyl-1-propyl (2S) -1- (1, 2-dioxo-2-phenyl) ethyl-2 -pyrrolidinecarboxylate, 99%, ¾ NMR (300 MHz , CDC13) : d 1.97-2.32 (m, SB); 2.74 (t, 2E, J = 7.5); '3.57 (m, 2H) ; 4.24 (m, 2H) ; 4.67 (m, IE); 6.95-7.28' (m, 5E) ; 7.51-7.64 (m, 3E) ; 8.03-8.09 (m, 2H) . Example 14: 3- (2, 5-dime hoxyphenyl) -1-propyl (2S) -1-(3 , 3 -dimethyl-1,2-dioxopentyl) -2-pyrrolidinecarboxylate, 99%, ¾ NM (300 MEz, CDC13) : d 0.87 (t, 3B) ; 1.22 (s, 3E) ; Ί.26 is, 3B); 1.69 (m, 2E) ; 1.96 (m, 5E) ; 2.24 (m, IE); 2.68 (m, 2E) ; 3.55 (m, 2E) ; 3.75 (s, 3E) ; 3.77 (s, 3E) ; 4.17 (m, 2E) ; 4.53 (d, IE); 6.72 (m, 3H) .

Example 15 : 3- (2, 5-dimethoxyphenyl) -l-prop-2-(E) -enyl (2S) -1- (3, 3 -dimethyl-1,2-dioxopentyl) -2-pyrrolidine-carboxylate, 99%, X NMR (300 MEz, CDC13) : d 0.87 (t, 3E) ; 1.22 (5, 3H) ; 1.26 (s, 3E) ; 1.67 (m, 2H) ; 1.73 (m, IE); 2.07 (m, 2H) ; 2.26 (m, IE); 3.52 (m, 2E) ; 3.73 (s, 3E) ; 3.80 (s, 3H) ; 4.54 (m, IE); 4.81 (m, 2E) ; 6.29 (dt, IH, u = 15. S); 6.98 (s, IH) .

Example IS : 2- (3 , 4, 5-trimethox phenyl) -1-ethyl (2S) -l- (3 , 3 -dimethyl-1, 2-dioxopentyl) -2-pyrrolidinecarboxylate, 97%, ¾ NMR (300 MHz, CDC13) : d 0.84 (t, 3H) ; 1.15 (s, 3H) ; 1.24 (s, 3H) ; 1.71 (dm, 2H);' 1.98 (m, 5H) ; 2.24 (m, IE); 2.63 (m, 2E) ; 3.51 (t, 2H) ; 3.79 (S, 3H) ; 3.83 (s, 3H) ; 4.14 (m, 2Ξ) ; 4.52 (m, IK); 6.36 (s, 2H) .

Example 17: 3 - (3 -Pyridyl) -1-propyl (2S) -1- (3,3-dimethyl-1, 2-dioxopentyl) -2-pyrrolidinecarboxylate, . 80%, ¾ M (CDClj, 300 MHz): d 0.85 (t, 3H) ; 1.23, 1.26 (s, 3H each) ; 1.63-1.89 (m, 2E) ; 1.90-2.30 (m, '■ 4H) ; 2.30-2.50 (m, IH) ; 2.72 (t, 23); 3.53 (m, 2H) ; 4,19 (m, "2E) ; 4.53' (m, IH) ; 7.22 (m, IH) ; 7.53 (dd, IH); 8.45.

Example 18: 3- (2 -Pyridyl) -1-propyl (2S) -1- (3 , 3-dimethyl-1, 2-dioxopentyl) -2-pyrrolidinecarboxylate, 88%, ¾ NMR (CDClj, 300 MHz) : d 0.84 (t, 3H) ; 1.22, 1.27 (s, 3H each); 1.68-2.32 (m, 8H) ; 2.88 (t, 2H, J = 7.5); 3.52 (m, 2H) ; 4.20 (m, 2H); 4.51 (m, IH) ; 7.09- 7.19 (m, 2H) ; 7.59 (m, IH) ; 8.53 (d, IH, J = 4.9).

■Example 19: 3- (4-Pyridyl)-l-propyl (2S) -1- (3 , 3-dimethyl-1, 2-dioxopentyl) -2-pyrrolidinecarboxylate, 91%, lH NMR (CDC13, 300. MHz): d 6.92-6.80 (m, 4H) ; 6.28 (m, IH) ; 5.25 (d, IH, J = 5.7); 4.12 (m, IE); 4.08 (s, 3H) ; 3.79 (s, 3H) ; 3.30 (m, 2H) ; 2.33 (m, IE); 1.85-1.22 (m( 7H) ; 1.25 (s, 3H) ; 1.23 (s, 3H) ; 0.89 (t, 3E, J = 7.5) .

Example 20: 3 -phenyl-I-propyl (2S) - I - (2-cyclo exyl-l, 2-dioxoethyi) -2-pyrroiidinecarboxylata, 91%, ¾ NMR (CDC13, 300 MHz): d 1.09-1.33 (m, SK) ; 1.62-2.33 (m, 12H) ; 2 '.€9 )t, 2H, J = 7.5); 3.15 (cm, 1H) ; 3.68 (m, 2K) ; 4.15 (m, 2E) ; 4.53, 4.84 {d, 1H total) ; 7.19 (m, 3H) ; 7.29 (m, 2K) .

Example 21: 3 -phenyl-1-propyl (2S) -1- (2- tert-butyl-1, 2-dioxoethyl) -2-pyrrolidinecarboxylate, 92%, lH NMR (CDClj, 300 MHz.) : d 1.29 (s, 9H) ; 1.94-2.03 (m, 5H) ; 2.21 (m, IE); 2.69 (m, 2H) ; 3.50-3.52 (m, 2H) ; 4.16 (m, 2H) ; 4.53- (m, 1H) ; 7.19 (m, 3H) ; 7.30 (m, 2H) .

Example 22: 3 -phenyl-1-propyl (2S) -1- (2-cyclohexyl-ethyl-1, 2-dioxoethyl) -2-pyrrolidinecarboxylate, 97%, ¾ NMR (CDCI3, 300 MHz): d 0.88 (m, 2K) ; 1.16 (m, 4H) ; 1.43-1.51 (m, 2H); 1.67 (m, 5H) ; 1.94-2.01 (m, 6H) ; 2.66-2.87 (m, 4H) / 3.62-3.77 (m, 2H) ; 4.15 (m, 2H) ; 4.86 (m, 1H) ; 7.17-7.32 (m, 5H) .

Example 23 : 3- (3-pyridyl) -1-propyl (2S) -1- (2-cyclo-hexylethyl-1, 2-dioxoethyl) -2-pyrrolidinecarboxylate, 70%, ¾ NMR (CDCI3, 300 MHz): d 0.87 (m, 2H) ; 1.16 (m, 4H) ; 1.49 (m, 2H) / 1.68 (m, 4H) ; 1.95-2.32 (m, 7H) ; 2.71 (m, 2H) ; 2.85 (m, 2H) ; 3.63-3.78 (m, 2H) / 4.19 (m, 2H) ; 5.30 (m, 1H) ; 7.23 (m, 1H) ; 7.53 (m, 1H)' ; 8 ]46 (m, 2H) .

Example 24: 3 - (3 -pyridyl) -1-propyl (2S) -1- (2- tert-butyl-1, 2-dioxoethyl) -2-pyrrolidinecarboxylate, S3%, ¾ NMR (CDCI3 , 300 MHz): d 1.29 (s, 9H) ; 1.95-2.04 (m, 5H) ; 2.31 (m, 1H) ; 2.72 (t, 2H, J = 7.5); 3.52 (m, 2H) ; 4.18 (m, 2Ξ) ; 4.52 (m, 1H) ; 7.19-7.25 (m, IE); 7.53 (m, 1H) ; 8.45 (m, 2H) .

Example 25: 3,3-dip enyl-l-propyl (2S) -1- (3 , 3 -dimethyl-1, 2-dioxoper.tyl) -2-pyrrolidinecarboxylate, 99%, ¾ NMR (CDC13, 300 MHz) : d 0.85 (t, 3H) ; 1.21,. 1.25 (s, 3K each); 1.68-2.04 (m, 5H) ; 2.31 (m, 1H) ; 2.40 (m, 2H) ; 3.51 (m, 2E) ; 4.08 (m, 3E); 4.52 (m, IE); 7.18-7.31 (m, lOE) .

Example 2S: 3 - (3 -pyridyl) -1-propyl (2S) -l- (2-cyclo-hexyl-1, 2-dioxoe.thyl) -2-pyrrolidinecarboxylate, 88%, ¾ NMR (CDCij, 300 MHz): d 1.24-1.28 (m, 5H) ; 1.88-2.35 (m, HE); 2.72 (t, 2E, J = 7.5); 3.00-3.33 (dm, IE); 3.63 (m, 2H) ; 4.13 (m, 2S) ; 4.55 (m, IE); 7.20-7.24 (m, IK); 7.53 (m, IE); 8.47 (m, 2H) .

Example 27: 3- (3-Pyridyl) -1-propyi (2S) -N- { [2-thienyl] glyoxyl) pyrrolidinecarboxylate, 49%, ¾ NMR (CDC13, 300 MHz): d 1.81-2.39 (m, SE) ; 2.72 (dm, 2H) ; 3.73 (m, 2E) ; 4.21 (m, 2E) ; 4.95 (m, IE); 7.19 (m, 2E) ; 7.61 (m, IE); 7.80 (d, IE); 8.04 (d, IE) ; 8.46 (m, 2E) .

Example 28: 3,3-Diphenyl-l-propyl 2S) -1- (3 , 3 -dimethyl- 1,2-dioxobutyl) -2-pyrrolidinecarboxylate, 99%, ¾ NMR (CDCI3, 300 MHz) : d 1.27 (s, 9E) ; 1.96 (m, 2Ξ) ; 2.44 (m, 4H) ; 3.49 (m, IE); 3.64 (m, IE); 4.08 (m, 4ΕΓ; 4.53 (dd, IE)" ; 7.24 (m, 10E) .

Example 29: 3,3-Diphenyl-l-propyl (2S) -1-cyclohexyl glyoxyl -2-pyrrolidinecarboxylate, 91%, ¾ NMR (CDC13, 300 MHz): d 1.32 (m, 6E) ; 1.54-2.41 (m, 10H) ; 3.20 . (dm, IE); 3.69 (m, 2H) ; 4.12 (m, 4E) ; 4.52 (d, IE); 7.28 (m, lOH) .

Example 30: 3 , 3 -Diphenyi-l-propyl (25) -l- (2-thienyl) glyoxyl-2-pyrrolidinecarboxylate, 75%, ¾ N R (CDC13, 300. MHz) : d 2.04 (m, 3K) ; 2.25 (m, 2H) ; 2.48 (m, IE) ; 3.70 (m, 2H) ; 3.82-4.18 (m, 3H total) ; 4.64 (m, IE) ; 7.25 (m, 11H).; 7.76 (dd, IE) ;· 8.03 (m, IE) .

The requisite substituted alcohols may be prepared by a number of methods known to those skilled in the art of organic synthesis.. As described in Scheme II, alkyl or -aryl aldehydes may be homologated to phenyl propanols by reaction with methyl (triphenylphosphoranylidene) acetate to provide a variety of trans-cinnamates ; these latter may be reduced to the saturated alcohols by reaction with excess lithium aluminum hydride, or sequentially by reduction of the double bond by catalytic hydrogenation and reduction of the saturated ester by appropriate., reducing agents. Alternatively, the trans-cinnamates may be reduced to (E) -allylic alcohols by the use of diisobutylaluminum hydride.

Longer chain alcohols may be prepared bv homologation of benzylic and higher aldehyd s.

Alternatively, these aldehydes may be prepared by conversion of the corresponding phenylacetic and higher acids, and phenethyl and higher alcohols.

General' procedure, for the synthesis of acrylic esters, exemplified for methyl (3 , 3 , 5-trimethoxy) -trans-cinnamate : A solution of 3 , 4, 5-trimethoxybenzaldehyde (5.0 g; 25.48 mmol) and methyl (triphenyl-phosphoranylidene) acetate (10.0 g; 29.91 mmol) in tetrahydrofuran (250 mL) was refluxed overnight. After cooling, the reaction mixture was diluted with.200 mL of ethyl acetate and washed 'with 2 x 200 mL of water, dried, and concentrated in vacuo. The crude residue was chromatographed on a silica gel column, eluting with 25% ethyl acetate in hexane, to obtain 5. S3 g (88%) of the cinnamate as a white crystalline solid, XH NM (300 Mhz; CDCLj) : d 3.78 (s, 3H) ; 3.85 (s, SH); 6.32 (d, 1H, J IS) ; S.72 (s, 2H) ; 7.59. (d, 1H, J = IS) .

General procedure for the synthesis of saturated alcohols from acrylic esters. Exemplified for (3,4,5-trimethoxy) phenylpropanol .

A solution of methyl (3 , 3 , 5-trimethoxy) - trans-cinnamate (1.81 g; 7.17 mmol) . in tetrahydrofuran (30 mL) was added in a dropwise manner to a solution of . lithium aluminum hydride (14 mmol) in THF (35 mL) , with stirring and under an argon atmosphere. After the addition, was complete, the mixture was heated to 75°C for 4 hours. After cooling, it was quenched by the careful addition of 15 mL of 2N NaOH followed by 50 mL of water. The resulting mixture was filtered through Celite to remove solids, and the filter cake was washed with ethyl acetate. The combined organic fractions were washed with water, dried, concentrated in vacuo, and purified on a silica gel column, eluting with ethyl acetate to obtain 0.86 g (53%) of the alcohol as a clear oil, lH NMR (300 MhZ; CDC13) : d 1.23 (br, 1H) ; 1.87 (m,'2H); 2.61 (t, 2H, J = 7.1); 3.66 (t, 2H) ; 3.80 (S, 3H) ; 3.83 (s, 6H) ; 6.40 (s, 2H) .

General procedure for the synthesis of trans-allylic alcohols from acrylic esters. Exemplified for (3,4, 5-trimethoxy)phenylprop-2- (Ξ) -enol.

A solution of methyl (3 , 3 , 5-trimethoxy) - trans--cinnamate (1.35 g; 5.35 mmol) in toluene (25 mL) was cooled to -10°C and treated with a solution of diisobutylaluminum hydride in toluene (11.25 mL of a 1.0 M solution; 11.25 mmol). The reaction mixture was stirred for 3 hrs at 0°C and then quenched with 3 mL of methanol followed by 1 HCl until the pH was 1. The reaction mixture was extracted into ethyl acetate" and the organic phase was washed with water, dried and concentrated-. Purification on a silica gel column eluting with 25% ethyl acetate in hexane furnished 0.95 g (80%) of a thick oil, ¾ MR (350 Mhz; CDC13) : d 3.85 (s, 3E) ;. 3.87 (s, 6H) ; 4.32 (d, 2H, J = 5.6); 6.29 {άΰ, IE, - - 15.8, 5.7), 6.54 (d, IE, J = 15.3); 6.61 (s, 2H) .

The invention being thus described, it will be obvious that the same may be varied in many ways. Such variations are not to be regarded as a departure from '■ the spirit and scope of the invention and all such modification are intended to be included within the scope of the following claims .

* * * * * Passages of the description which are not within the scope of the claims do not form part of the invention.

Claims (8)

134562/2 52 C L A I M S:

1. A compound of the formula: where Ri is a C1-C9 straight or branched chain alkyl or C2-C9 straight or branched alkenyl group optionally substituted with C3-Cs cycloalkyl; C3 or C5 cycloalkyl, C3-C7 cycloalkenyl, or Ar:, where aforementioned alkyl, alkenyl , cycloalkyl, or cycloalkenyl groups may optionally have one or more hydrogen atoms replaced with Ca-C4 alkyl, C2-C4 alkenyl, or hydroxy, and where hrx is selected from the group consisting of 1-naphthyl, 2-naphthyl, 2-indolyl, 3-indolyl, 2-furyl, 3-furyl, 2-thiazolyl, 2-thienyl, 3-thienyl, 2-, 3-, or 4-pyridyl, and phenyl, having' one to three substituents which are independently selected from the group consisting of hydrogen, halogen, hydroxyl, nitro, trif luoromethyl, Ci-Cs straight or branched alkyl or C2-C6 straight or branched alkenyl, C C4 alkoxy or C2-C4 alkenyloxy, phenoxy, benxyloxy, and amino; or pharmaceutically acceptable salts or hydrates thereof .

2. . The compound of claim 1, which has an affinity for FKBP-type immunophilins . 53 134562/4

3. The compound of claim 2, where the FKBP-type immunophilin is F BP-12.

4. The compound of claim 1, capable of inhibiting rotamase activity.

5. The compound of claim 1, where R\ is a lipophilic group.

6. A compound that is (2S)-l-(l,2-dioxo-3,3-dimethylpentyl)-2-pyrrollidine carboxylic acid.

7. A method of synthesizing a chemical compound according to Claim 6, comprising: forming a mixture of methyl (2S)-l-(l,2-dioxo-3,3-dimethylpentyl)-2- pyrrollidinecarboxylate, IN lithium hydroxide, and methanol; stirring the mixture at 0°C for 30 minutes and at room temperature for about 12 hours; acidifying with IN hydrochloric acid to pH 1 ; diluting with water; and extracting the organic phase into methylene chloride.

8. A compound according to claims 1 or 6 of general formula as herein described and pharmaceutically acceptable salts thereof, substantially as hereinbefore described with reference the accompanying examples. For the Applicants REINHOLD COHN AND PARTNERS By: