GB2325230A - Intermediate for Neurotrophic N-glyoxylprolyl esters - Google Patents

Abstract

(25)-1-(1,2-dioxo-3,3-dimethylpentyl)-2-pyrrolidinecarboxylic acid and its preparation are claimed.

Description

2325230 SMALL MOLECULE INHIBITORS OF ROTAMASE ENZYME ACTIVITy Related

Application This application is a continuation-in-part application of U.S. Patent Application Serial No.

08/479,436 filed June 7, 1995.

BACKGROUND OF THE INVENTION

I. Field of the Invention

This invention relates to neurotrophic compounds having an affinity for FKBP-type immunophilins, their preparation and use as inhibitors of the enzyme activity associated with immunophilin proteins, and particularly inhibitors of peptidyl-prolyl isomerase or rotamase enzyme activity.

2. Description of the Prior Art

The term immunophilin refers to a number of proteins that serve as receptors for the principal immunosuppressant drugs, cyclosporin A (CsA), FK506, and rapamycin. Known classes of immunophilins are cyclophilins, and FK506 binding proteins, such as FKBP.

Cyclosporin A binds to cyclophilin while FK506 and rapamycin bind to FKBP. These immunophilin-drug complexes interface with a variety of intracellular signal transduction systems, especially in the immune system and the nervous system.

Tmmunophilins are known to have peptidyl-prolyl 2 isomerase (PPIase) or rotamase enzyme activity. It has been determined that rotamase activity has a role in the catalyzation 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-cell proliferation, thereby causing the immunosuppressive action exhibited by immunosuppressive drugs such as cyclosporin A, FK506, 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. 556-559. It has been shown that the immunoPhilin-drug complexes interact with ternary protein targets as their mode of action. Schreiber et al., Cell, 1991, vol. G6, pp. 807-815. In the case of FKBP-FK506 and FKBP-CsA, the drug-immunophilin complexes bind to the enzyme calcineurin, inhibitory T- cell receptor signalling leading to T-cell prol iferation. Similarly, the complex of rapamycin and FKBP interacts with the P-AFT1/FRA.P 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.

3 Within neural tissues, immunophilins appear to influence neuronal process extension, nitric oxide synthesis, and neurotransmitter release.

It has been found that picomolar concentrations of an immunosuppressant such as FKSOG and rapamycin stimulate neurite out growth in PC12 cells and sensory nervous, namely dorsal root ganglion cells (DRGs) Lyons et al. , Proc. of Natl. Acad. Sci., 1994 vol. 91, pp. 3191-3195. In whole animal experiments, FK506 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 neurolegenerative disorders such as Alzheimer's disease, Parkinson's disease, and amyotrophic lateral sclerosis (ALS) may occur due to the loss, or 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 a loss of nerve growth factor (NGF) 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 is chronically, immunosuppressants exhibit a number of potentially serious side effects including nephroto:icity, such as impairment of glomerular filtration and irreversible interstitial fibrosis (Kopp et al., 1991, J. Am. Soc. Nephrol.

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 non - immuno suppressive 1:162).. neurolocrical compounds containing small molecule FKBP 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 THE INVENTION

The present invention relates to a novel class of neurotrophic compounds having an affinity for FKBP-type is immunophilins. Once bound to this protein the neurotrophic compounds are patent 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 s ignificant immuncsuppressive activity in addition to their neurotrophic activity.

A preferred embodiment of this invention is a neurotrophic compound of the formula:

6 _Z N. 1 0 0)--- X R' 1 where is R, is selected from the group consisting of a C,-C9 straight or branched chain alkyl or alkenyl group optionally substituted with C3-C, cycloalkyl, C, or C. cycloalkyl, C5C, cycloalkenyl, Arl, where said alkyl, alkenyl, cycloalkyl or cycloalkenyl groups may be optionally substituted with C,-C, alkyl, Cl C4 alkenyl, or hydroxy, where Ar, is selected from the group consisting of l-napthyl, 2-napthyl, 2 indolyl, 3-indolyl, 2-furyl, 3-furyl, 2-thiazolyl, 2-thienyl, 3-thienyl, 2-,3-, 4-pyridyl, and phenyl, having one to three substituents which are independently selected from the group consis"ting of hydrogen, halo, hydroxyl, nitro, trifluoromethyl, Cl-C6 straight or branched alkyl or alkenyl, Cl-C, alkoxy or Cl-C, alkenyloxy, phenoxy, benzyloxy, and amino; X is selected from the croup consisting of 7 oxygen, sulfur, methylene (CH 2), or H2; Y is selected from the group consisting of oxygen or NR2, where R2 is hydrogen or Cl alkyl; and is selected from the group consisting of C2 C6 straight or branched chain alkyl or alkenyl, wherein the alkyl chain is substituted in one or more positions with Ar, as defined above, C3-C, cycloalkyl, cycloalkyl connected by a Cl-C, straight or unbranched alkyl or alkenyl chain, and Ar2 where Ar2 is selected from the group consisting of 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, halo, hydroxyl, nitro, trifluoromethyl, Cl-C,, straight or branched alkyl or alkenyl, Cl-Q, alkoxy or Cl-C, alkenyloxy, phenoxy, benzyloxy, and amino; Z may also be the fragment:

is _C6 z 0 -CH 11 X -R 2 4 R 3 1 f 8 where R3 is selected from the group consisting of straight or branched alkyl C,-Ca optionally substituted with C3-C, cycloalkyl, or Ar, as defined above, and unsubstituted Ar,; X2 is 0 or NR,, where R. is selected from the group consisting of hydrogen, Cl-C6 straight or branched alkyl and alkenyl; R4 is selected from the group consisting of phenyl, benzyl,, C,-C, straight or branched alkyl or alkenyl, and Cl-C, straight or branched alkyl or alkenyl substituted with phenyl; or pharmaceutically acceptable salts or hydrates thereof.

is Another preferred embodiment of this invention is a neurotrophic compound of the formula:

CN o-z 0 1.

R 0 1 9 where 1 z I- R, is a Cl-C, straight or branched chain alkyl or alkenyl group optionally substituted with C3-C, cycloalkyl, C3 or C5 cycloalkyl, C S-C7 cycloalkenyl, or Arl, where said alkyl, alkenyl, cycloalkyl or cycloalkenyl groups may be optionally substituted with c 1-C4 alkyl, Cl-C, alkenyl, or hydroxy, and where Ar, is selected from tkie group consisting of l-napthyl, 2-napthyl, 2- indolyl, 3-indolyl, 2-furyl, 3-furyl, 2-thiazolyl, 2-thienyl, 3-thienyl, 2-,3-, or 4-pyridyl, or phenyl, having one to t.hree substituents which are independently selected from the group is consisting of hydrogen, halo, hydroxyl, nitro, trifluoromethyl, Cl-C, straight or branched alkyl or alkenyl, Cl-C, alkoxy or ClC, alkenyloxy, phenoxy, benzyloxy, and amino; is a C2-C, straight or branched chain alkyl or alkenyl, wherein the alkyl chain is substituted in one or more positions with Ar, as defined above, C3-C, cycloalkyl, cycloalkyl connected by a Cl-C, straight. or unbranched alkyl or alkenyl chain, or Ar2 where Ar. is selected from the group consisting of 2-indolyl, 3-indolyl, 2-furyl, 3-furyl, 2- thiazolyl, 2-thienyl, 3thienyl, 2-, 3-, or 4pyridyl, or phenyl, having one 1 i " to three substituents which are independently selected from the group consisting of hydrogen, halo, hydroxyl, nitro, trifluoromethyl, CI-C6 straight or branched alkyl or alkenyl, C,-C, alkoxy or Cl-C, alkenyloxy, phenoxy, benzyloxy, and amino; or pharmaceutically acceptable salts or hydrates thereof.

Another preferred embodiment of the invention is a neurotrophic compound having an affinity for FKBP-ty-oe immunophilins which inhibit the rotamase activity of the immunophilin.

Another preferred embodiment of the present Jnvention is a method for treating a neurological is disorder in an animal comprising administering a therapeutically effective amount of a compound having an affinity for FKBP-type immunophilins which inhibits the rotamase activity of the immunophilin.

Another preferred embodiment of the invention is a method of promoting neuronal regeneration and growth in mammals, comprising administering to a mammal an effective amount of a neurotrophic compound having an affinity for FKBP-type immunophilins which inhibits the rotamase activity of the immunophilin.

Yet another preferred embodiment of the invention is a method of preventing neurodegeneration in an animal comprising administering to an animal an effective amount of a neurotrophic compound having an 11 affinity for FKBP-type immunophilins which inhibits rotamase activity of the immunophilin.

Another preferred embodiment is a neurotrophic N glyoxyl prolyl ester compound of the formula:

N 0- 0, 1 ll ' R is where X Y z z R, is a C,-C, straight or branched chain alkyl or alkenyl group optionally substituted with C3 to C6 cycloalkyl, or Arl, where Ar, is selected from the group consisting of 2furyl, 2-thienyl, or phenyl; is selected from the group consisting of oxygen and sulfur; is oxygen; and is a straight or branched chain alkyl or alkenyl, wherein the alkyl chain is substituted in one or more positions with Ax, as defined above, C3-C, cycloalkyl, Ar2 where 12 Ar2 'S selected from the group consisting of 2-, 3-, or 4-pyridyl, or phenyl, having one to three substituents which are independently selected from the group consisting of hydrogen and Cl-C4 alkocy.

Particularly preferred neuratrophic N-glyoxyl prolyl ester compounds according to the above formula are selected from the group consisting of:

3-(2,5-dimethoxyphenyl)-1-propy1 (2S) -1- (3,310 dimezhyl-1,2dioxopentyl)2-pyrrolidinecarboxylate, 3-(2,5-dimethoxyphenyl)-1-prop-2-(E)-enyl (2S)-1(3,3-dimethyl-1,2dioxopentyl)-2-pyrrolidinecarboxylate, 2 (3,4, 5 -trimethoxyphenyl) - 1 -ethyl (2S)-1-(3,3is dime--hyl-1,2- dioxopentyl) -2-pyrrolidinecarboxylate, 3-(3-Pyridyl)-1-propyl (2S)-1-(3,3-dimethyl-1,2dioxopentyl)-2- pyrrolidinecarboxylate, 3-(2-Pyridyl)-1-propyl (2S)-1-(3,3-dimethyl-1,2dioxopentyl)-2pyrrolidinecarboxylate, 3-(4-Pyridyl)-1-propyl (2S)-1-(3,3-dimethyl-1,2dioxopentyl)-2-pyrrolidinecarboxylate, 3 -phenyl - 1-propyl (2S)-1-(2-tert-bu'tyl-1,2dioxoethyl)-2- pyrrolidinecarboxylate, 3-phenyl-l-propyl (2S)-1-(2-cyclohexylethyl1,225 dioxcethyl)-2pyrrolidinecarboxylate, 3-(3-pyridyl)-1-propyl (2S)-1-(2-eyelchexylethyl1,2-di-oxoethyl)-2- pyrralidinecarboxylate, 3-(3-pyridyl)-1-propyl (2S)-1-(2-tert-but:yl-1,2- 0 13 dioxoethyl)-2-pyrrolidine carboxylate, 3,3-diphenyl-l-propyl (2S)-1-(3,3-dimethyl-1,2- dioxopentyl)-2-pyrrolidinecarboxylate, 3-(3-pyridyl)-1-propyl (2S)-1-(2-cyclohexyl-1,2dioxoethyl)-2-pyrrolidinecarboxylate, 3(3-Pyridyl)-1-proDyl (2S)-N-([2-thienyll glyoxyl)pyrrolidinecarboxylate, 3,3-Diphenyl-l-propyl (2S)-1-(3,3-dimethyl-1,2dioxobutyl)-2-pyrrolidinecarboxylate,,3-Diphenyl-l-propyl (2S)-1cyclohexylglyoxyl2-pyrrolidinecarboxylate, and 3,3-Diphenyl-l-propyl (2S)-1-(2-thienyl)glyoxyl- 2-pyrrolidinecarboxylate.

BRIEF DESCRIPTION OF THE DRAWINGS

Figure I is a photomicrograph of chick dorsal root ganglia treated with various concentrations of Exaiuple 17 as indicated. Figure I 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.

14 Micrographs of Example 17 treated DRG's, as well as quantitative dosedependent 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. Fig-ure 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 17treated samples to yield Example 17-dependent specific neurite outgrowth. Quantitative dose-dependent neurite outgrowth elicited by Example 17 is presented.

Fig-ure 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 is crush site were sectioned and stained with Holmes silver stain (to assess axon number) and Luxol fast blue (to assess remyelination). The micrographs show sciatic nerve sections of sham operated rats, vehicle- treated lesioned animals, Example 1 and Inactive treated at 630x magnification, four animals per group.

Figure 4 is a graph of [ 3 H] -CFT binding per ug of Striatal Membrane Protein. Figure 4 shows that neuraimmunophilin ligands of the present invention promote recovery of dopamine neurons following MPTP 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 is (100 mg/kg s.c.) or Example 17 (40, 20, 10 mg/kg s.c., as indicated) c9ncurrently with the MPTP 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 E3HI-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 10 gM unlabelled CFT provided on estimate of nonspecific binding, which was subtracted from the total binding to quantitative specific [3HI-CFT bound. Binding was normalized to the protein content of the striatal membranes from each experimental group. Coronal and 16 saggital brain sections from MPTP and drug treated animals were stained with anti-tyrosine hydroxylase (TH) Ig to quantitate striatal, medial forebrain bundle axonal and nigral levels of TH, which is 5 indicative of functional dopaminergic neurons.

Figure 5 is a bar graph of [3H] -CFT plotted f or 200 gg of membrane protein. Figure 5 shows that neuroimmunophilin ligandg of the present invention promote recovery of dopamine neurons following MPTP treatment of mice in accordance with the procedure described in Figure 4.

Figure 6 is a photomic'rograph, at 630x magnification, of coronal and saggital brain sections. Figure 6 shows brain sections from MPTP and drug is treated animals stained with anti-tyrosine hydroxylase (TH) Ig to quantitate striatal levels of TH, which is indicative of functional dopaminergic neurons.

Figure 7 is a photomicrograph, at SOx magnification, of coronal and saggital brain sections.

Figure 7 shows brain sections from MPTP and drug treated animals stained with anti-tyrosine hydroxylase (TH) Ig to quantitate nigral levels of TH, which is indicative of functional dopaminergic neurons.

Figure 8 is a photomicrograph, at 400x magnification, of coronal and saggital brain sections. Figure 8 shows brain sections from MPTP and drug treated animals stained with anti- tyrosine hydroxylase (TH) Ig to quantitate medial forebrain bundle axonal 17 levels of TH, which is indicative of functional dopaminergic neurons.

DETAILED DESCRIPTION OF THE INVENTION

The novel neurotrophic compounds of this invention are relatively small molecules in relation to other known compounds which bind to FKBP-type immunophilins, such as rapamycin, FK506, and cyclosporin.

The neurotrophic compounds of this invention have an affinity for the FK506 binding proteins such as FKBP-12. When the neurotrophic compounds of the invention are 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 neurite growth, while not exhibiting an immunosuppressant effect.

More particularly, this invention relates to a novel class of neurotrophic compounds represented by the formula:

-Z N 1 0 0)--- X R' 18 I where j is X Y is a Cl-C9 straight or branched chain alkyl or alkenyl group optionally substituted with C3-C, cycloalkyl, C3 or C5 cycloalkyl, C,-c. cycloalkenyl, or Arl, where said alkyl, alkenyl, cycloalkyl or cycldalkenyl groups may be optionally substituted with C,-C. alkyl, C,-C, alkenyl, or hydroxy, and where Ar, is selected from the group consisting of l-napthyl, 2-napthyl, 2-indolyl, 3-indolyl, 2-furyl, 3-furyl, 2- thiazolyl, 2-thienyl, 3thienyl, 2-, 3-, or 4-pyridyl, or phenyl, having one to three substituents which are independently selected from the group consisting of hydrogen, halo, hydroxyl, nitro, trifluoromethyl, Cl-C, straight or branched alkyl or alkenyl, Cl-C, alkoxy or Cl C, alkenyloxy, phenoxy, benzyloxy, and amino is oxygen, sulfur, methylene (CH7), or H2; is oxygen or NR2. where R. is hydrogen or ClC. alkyl; and is a C2-C, straight or branched chain alkyl or alkenyl, wherein the alkyl chain is substituted in one or more positions with Ar as defined above, C3-CS cycloalkyl, cycloalkyl connected by a C,-C, straight or where 19 unbranched alkyl or alkenyl chain, or 'IX2 where Ar2 is selected from the group consisting of 2-indolyl, 3-indolyl, 2furyl, 3-furyl, 2thiazolyl, 2-thienyl, 3thienyl, 2-, 3-., or 4-pyridyl, or phenyl, having one to three substituents which are independently selected from the group consisting of hydrogen, halo, hydroxyl, nitro, trifluoromethyl, Cl-C, straight or branched alkyl or alkenyl, Cl-C, alkoxy or Cl-C4 alkenyloxy, phenoxy, benzyloxy, and amino; may also be the fragment:

0 CH 11 X2 -R 1 - 4 R 3 is selected from the group consisting of straight or branched alkyl Cl-C, optionally substituted with C3-C8 cycloalkyl, or Ar, as defined above, and unsubstituted Arl; X2 is 0 or NR., where R. is selected from the i is group consisting of hydrogen, C-I-C6 straight or branched alkyl and alkenyl; R4 is selected from the group consisting of phenyl, benzyl, C,-C. straight or branched alkyl or alkenyl, and C,-Cs straight or branched alkyl or alkenyl substituted with phenyl; or pharmaceutically acceptable salts or hydrates thereof.

Preferred compounds -have the following formula:

c 0-Z N 1 0 0 R where II R, is a C,-C, straight or branched chain alkyl or alkenyl group optionally substituted with C,-C, cycloalkyl, C, or C. cycloalkyl, C5-C, cycloalkenyl, or Arl, where said alkyl, alkenyl, cycloalkyl or cycloalkenyl groups may be optionally substituted with C,-C, is 21 alkyl, C.-C, alkenyl, or hydroxy, and where Arl is selected from the group consisting of 1-napthyl, 2-napthyl, 2-indolyl, 3-indolyl, 2-furyl, 3-furyl, 2- thiazolyl, 2-thienyl, 3thienyl, 2-, 3-, or 4-pyridyl, or phenyl, having one to three substituents which are independently selected from the group consisting of hydrogen, halo, hydroxyl, nitro, trifluoromethyl, Cj-C6 straight or branched alkyl or alkenyl, Cl-C, alkoxy or C,C, alkenyloxy, phenoxy, benzyloxy, and amino; is a C.-C. straight or branched chain alkyl or alkenyl, wherein the alkyl chain is substituted in one or more positions with Ar, as defined above, C3-C8 cycloalkyl, cycloalkyl connected by a C,-C. straight or unbranched alkyl or alkenyl chain, or Ar. where Ar2 is selected from the group consisting of 2-indolyl, 3-indolyl, 2furyl, 3-furyl, 2- thiazolyl, 2-thienyl, 3thienyl, 2-, 3-, or 4-pyridyl, or phenyl, having one to three substituents which are independently selected from the group consisting of hydrogen, halo, hydroxyl, nitro, trifluoromethyl, CI-C, straight or branched alkyl or alkenyl, C,-C, alkoxy or CjC. alkenyloxy, phenoxy, benzyloxy, and amino; or pharmaceutically acceptable salts or -z c 22 hydrates thereof.

Preferred neurotrophic N-91YOxyl prolyl ester compounds have the formula:

CN- 0-Z 1 OY 111 0 1 R where is R, X Y z is a C,-C, straight or branched chain alkyl or alkenyl group optionally substituted with C3 to C6 cycloalkyl, or Ar,, where Ar, is selected from the group consisting of 2fur-yl, 2-thienyl, or phenyl; is selected from the group consisting of oxygen and sulfur; is oxygen; and 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, C,_C6 cycloalkyl, Ar2 where Ar2 'S selected from the group consisting of i 23 2-, 3-, or 4-pyridyl, or phenyl, having one to three substituents which are independently selected from the group consisting of hydrogen and Cl-C, alkoxy.

1 1 The compounds of this invention exist as stereoisomeric forms, either enantiomers or diastereoisomers. The st ereochemistry at position 1 (Formula 1) is R or S, with S 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 FKBP 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 Biochemistzy, 29, 3813-38'16. 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 R, groups which are not stereochemically bulky 2S in relation to the known shape and size of the hydrophobic core of the FKBP active site. Thus, very large and/or highly substituted R, groups would bind Y 24 with less affinity to the FKBP active site.

Preferred compounds of the invention include:

3 -phenyl - 1 -propyl (2S) - 1- (3, 3 -dimethyl - 1, 2 dioxopentyl) -2pyrrolidinecarboxylate, 3 -phenyl -1-prop-2- (E) -enyl (2S)-1-(3,3-dimethyl1, 2 -dioxopentyl) -2 - pyrrolidinecarboxylate, 3-(3,4,5-trimethox-yphenyl)-1-propy1 (2S)-1(3,3dimethyl - 1, 2 dioxopentyl) - 2 -pyrrolidinecarboxylate, 3-(3,4,5-trimethoxyphenyl)-1-prop-2-(E)-enyl (2S)- 1- (3, 3 -dimethyl - 1, 2 -dioxopentyl) - 2 -pyrrolidine carboxylate, 3-(4,5-methylenedioxyphenyl)-1-propyl (2S)-1-(3,3dimethyl-1, 2-dioxo, pentyl) -2-pyrralidinecarboxylate, 3 -'(4, 5-methylenedioxyphenyl) -1-prop-2- (E) -enyl (2 S) - 1 - (3, 3 - dime thyl - 1, 2 - dioxopentyl) 2 pyrrolidinecarboxylate, 3-cyclohexyl-1-propyl (2S)-1-(3,3-dimethyl1,2dioxopentyl) -2- pyrrolidinecarboxylat--e, 3-cyclohexyl-l-prop-2-(E)-enyl (2S)-1-(3,3dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate, (1R)-1,3-diphenyl-lpropyl (2S)-1-(3,3-dimethyl1, 2 -dioxopentyl) - 2 -pyrrolidinecarboxylate, 3 -phenyl- 1-propyl (2S)-1-(1,2-dioxo-2E2furanyll)ethyl -2 pyrrolidinecarboxylate, 3-phenyl-l-propyl (2S)-1-(1,2-dioxo-2-E2th-Jenyl ethyl - 2 - pyrrolidinecarboxylate, 3-phenyl-l-propyl (2S)-1-(1,2-dioxo-2-[2thiazolyll)ethyl-2- pyrrolidinecarboxylate, 1 1 3 -phenyl -1 -propyl (2S)-1-(1,2-dioxc)-2phenyl)ethyl-2pyrrolidinecarboxylate, 3 - (2, 5 -dime t hoxyphenyl) - 1 -propyl (2S) l- (3, 3 dimethyl - 1, 2 -dioxopentyl) - 2 -pyrrolidinecarboxylate 3 - (2, 5 -dime t hoxyphenyl) - 1 -prop - 2 - (E) -enyl (2S)- 1 (3, 3 -dimethyl 1, 2 -dioxopentyl) - 2 -pyrrolidine carboxylate, 2 - (3,4, 5-trimethoxyphenyl) - 1 -ethyl (2S)-1-(3,3dimethyl - 1, 2 - dioxopentyl) 2 -pyrrol idinecarboxylate, 3- (3-Pyridyl) 1-propyl (2 S) -l- (3, 3-dimethyl- l, 2dioxopentyl) -2- pyrrolidinecarboxylate, 3-(2-Pyridyl)-1-propyl (2S)-1-(3,3-dimethyl-1,2diaxopentyl) -2pyrrolidinecarboxylate, 3-(4-Pyridyl)-1-propyl (2S)-1-(3,3-dimethyl-1,2is dioxopentyl) -2- pyrrolidinecarboxylate, 3 -phenyl - 1-propyl (2S)-1-(2-cyclohexyl-1,2diaxoethyl) -2- pyrrolidinecarboxylate, 3 -phenyl -1-propyl (2S)-1-(2-t:ert:-butyl-1,2dioxoethyl) -2pyrrolidinecarboxylate, 3-phenyl-1-propyl (2S)-1-(2-eyclohexylethyl1,2dioxoethyl) -2- pyrrolidinecarboxylate, 3(3-pyridyl) -1-propyl '(2S) -l- (2-cyclohexylethyl1,2-dioxoethyl)-2pyrrolidinecarboxylate, 3-(3-pyridyl)-1-propyl (2S)-1-(2-tert:-butyl-1,2dioxoethyl) -2- pyrrolidinecarboxylate, 3,3-diphenyll-propyl (2S)-1-(3,3-dimethyl-1,2dioxopentyl) -2pyrrolidinecarboxylate, 3-(3-pyridyl)-1-propyl (2S)-1-(2-cyclohexyl-1,2- 1 26 diaxoethyl)-2-pyrrolidinecarboxylate, 3-(3-Pyridyl)-1-propyl (2S)N-(C2-thienyl] glyoxyl)pyrrolidinecarboxylate, 3,3-Diphenyl-1-propyl (2S)-1-(3,3-dimethyl-1,25 dioxobutyl)-2- pyrrolidinecarboxylate, 3,3-Diphenyl-1-propyl (2S)-1-cyclohexylglycxyl2-pyrrolidinecarboxylate, 3,3-Diphenyl-1-propyl (2S)-1-(2-thienyl)glyoxyl2-pyrrolidinecarboxylate.

Particularly preferred neurotrophic N-glyoxyl prolyl ester compounds are selected from the group consisting of:

3-(2,5-dimethoxyphenyl)-1-propyl (2S)-1-(3,3dimethyl-1, 2-dioxopentyl) -2-pyrrolidinecarboxylate, is 3-(2,5-dimethoxyphenyl)-1-prop2(E)-enyl (2S)-1(3,3-dimethyl-1,2-dioxopentyl)-2-pyrrolidinecarboxylate, 2-(3,4,5-trimethoxyphenyl)-1-ethyl (2S)-1-(3,3dimethyl-1,2-dioxopentyl)-2pyrrolidinecarboxylate, 3-(3-Pyridyl)-1-propyl (2S)-1-(3,3-dimethyl-1,2diaxopentyl)-2- pyrrolidinecarboxylate, 3-(2-Pyridyl)-1-propyl (2S)-1-(3,3-dimethyl-1,2dioxopentyl)-2pyrrolidinecarboxylate, 3-(4-Pyridyl)-1-propyl (2S)-1-(3,3-dimethyl-1,2dioxopentyl)-2-pyrrolidinecarboxylate, 3-phenyl-l-propyl (2S)-1-(2-t:ert-butyl-1,2dioxoethyl)-2- pyrrolidinecarboxylate, 3 -phenyl1-propyl (2S)-1-(2-eyclohexylethyl-1,2- 4 1 1 27 dioxoethyl)-2-pyrrolidinecarboxylate, 3-(3-pyridyl)-1-propyl (2S)-1-(2cyclohexylethyl1,2-dioxoethyl)-2- pyrrolidinecarboxylate, 3-(3-pyridyl)-1-propyl (2S)-1-(2-t:ez-t-butyl-1,25 dioxoethyl)-2pyrrolidinecarboxylate, 3,3-diphenyl-l-propyl (2S)-1-(3,3-dimethyl-1,2dioxopentyl)-2- pyrrolidinecarboxylate, 3-(3-pyridyl)-1-propyl (2S)-1-(2-cyclohexyl-1,2.dioxoethyl)-2pyrrolidinecarboxylate, 3-(3-PYridyl)-1-propiyl (2S)-N-(E2-thienyl] glyoxyl)pyrrolidinecarboxylate, 3,3-Diphenyl-1-propyl (2S)-1-(3,3-dimethyl-1,2dioxobutyl)-2- pyrrolidinecarboxylate, 3,3-Diphenyl-1-propyl (2S) -1-cyclohexylglyoxyl- 2-pyrrolidinecarboxylate, and 3,3-Diphenyl-l-propyl (2S)-1-(2-thienyl)glyoxyl2-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- is 1 28 hydroxyethanesulfonate, lactate, maleate, methane sul f onate, 2-naphthalensulfonate, 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, Nmethyl-Dglucamine, and salts with amino acids such'as arginine, lysine, and so forth. Also, the basic nitrogen-containing groups can be quarternized with such agents as lower alkyl halides, such as methyl, ethyl, propyl, and butyl chloride, 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 neironal regeneration and growth, such as in various peripheral neuropathic and neurological disorders relating to neurodegeneration. The compounds of this invention can also be administered to mammals other than humans for treatment of various mammalian neurological disorders.

29 The novel compounds cf: the present invention are poient inhibitors of' rotamase acc-.v--V and i:-)cssess an excellent degree of: neurotrophic ac---,v-Jtv. This activity is useful in the of damaged neurons, the promotion of neurona- -regeneration, -ne neurodegenera -1 p--eljeri--ion c JI the of se.veral neurological disorders known to be associated w-h neuronal degenerailo- and per-ic)"------=1 rieuropar-hies. The neurological disorders that may be treated include but are not ic: ', r -i 5 e 7 r,, -i---a- 17 neuralgia, glossopharyngeal neL,-a"ca, Bell's Pals,", r,,.-,ras--henia gravis, musculal- am, o lat:e---al sclerosis, progressive,muscular atrc-ohy, pr,:,gress-ive bulbar inherited musc---=r atrophy, J-ted, ri-l)Lured cr n e rn vertabrae disk sy---ndromes, ce--v-ica-! spondy"-jos-,s, disorders, thoracic outlet destruction sy-ndrommes, periphera-, neuroQathic such as those caused by lead, dapsone, prophyria, or Gullain-Barrd syndrome, Alzheimer's disease, and Parkinson's disease.

For these Purposes the compounds off the oresen-invention may be administered orally, parenterally, '1--y inhalation spray, Ciopically, rectally, nasally, buccally, vacjina-i-l-,,, or via an implanted reservoir dosage formulations containing conventional non-coxic pharmaceuticallyacceptable carriers, adjuvants and vehicles. The term parenteral as used herein includes subcutaneous, intravenous, intramuscular, intraperitoneally, intrathecally, intraventricularly, intrasternal and incracranial injection or infusion techniques.

To be effective therapeutically, as central nervous system targets the immunophilin-drug complex should readily penetrate the bloodbrain 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 S-Lerile injectable aqueous or cleag-inous 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 parenterallyacceptable diluent or solvent, for example as a solution in 1, 3-butanediol. Among the acceptable vehicles and 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 r-he-ir 31 polyoxyeLhylated 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 acrueous suspension or solution. In the case of tablets for oral use, carriers which are commonly used include 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 corn 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 recral 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 bur-ter, beeswax and polyethylene glycolsThe 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 is lower intestinal tract.. are readily prepared for For ophthalmic use, formulated as micronized adjust:ed sterile saline, is isotonic, pH adjusted 2 Suitable topical formulat-ions each of these areas. the compounds can be suspensions in isotonic, pp, or, preferably, as solutions stierile 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, a mixture with one or more of the following: mineral oil, liquid petrolatum, white petrolatum, is 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 monostearat-e, polysorbate 60, cetyl esters wax, cetearyl alcohol, 2octyldodecanol, benzyl alcohol and water.

Topical application for the lower intestinal-tract an be effected in a rectal suQoosit-ory formulation (see above) or in a suitable enema formulation. f about Img to Dosage levels on the orde-r o. lo,ooo mg. of the active ingredient compound are in the treatment of the above conditions, with 33 preferred levels of about O.1mg to about- 1,ooo mg. amount of 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. 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, rate of excretion, drug combination, and the severity of the particular disease being treated and form of administration.

The compounds can be administered with other 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.

KiTest Procedure inh-bition of the peptidyl-prolyl isomerase (rotamase) activity of the inventive compounds can be evaluated by known methods described in the literacure (Harding, M.W. ec a!. Nature 341: 758-760 (1989); H01: et al. j. Am. Chem. Soc. 115: 9923- 9938). These values are obcained as apparenr- Ki's and are presented in f 1 34 Table I. The cis-trans isomerization of an alanineproline bond in a model substrate, N-succinYl-Ala-AlaPro-Phe-p-nitroanilide, is monitored 1 1 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 Ki values.

In a plastic cuvette are added 950 mL of ice cold assay buf f er (2 5 mM HEPES, pH 7. 8, 10 0 mM NaCl), 10 mL of FKBP (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 1 mM HCI) and io 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- nitroanil ide, 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 ti-ne data files.

The data for these experiments is presented in Table I.

is 4 6 7 8 9 10 11 12 13 14 ' is 16 17 18 19 21 22 23 24 26 27 28 29 30 Table 1

Nr 0 '1 R 1 0 o\-\ 0 W No. R 1,1-dimethylpropyl I@ If 01 11 11 11 11 R 2-furanyl 2-thienyl 2-thiazolyl phenyl 1,1-dimethylpropyl 11 1 r 19 11 Il cyclohexyl tert-butyl cyclohexylethyl 01 cyclohexylethyl 3-(3-pyridyl)propyl tert-butyl 3-(3-pyridyl)propyl 1,1-dimethylpropyl 3,3-diphenylpropyl cyclohexyl 3-(3-pyridyl)propyl 2-thienyl 3-(3-pyridyl)propyl tert-butyl 3,3-diphenylpropyl cyclolmxyl 2-thienyl q@ R' 3-phenylpropyl 3-phenyl-prop-2- 125 (E)-enyl 3-(3,4,5-tri methoxyphenyl)propyl 200 3-(3,4,5-trimethoxy- phenyl)-prop-2-(E)-enyl 65 3-(4,5-methylenedioxy) phenylpropyl 3-(4,5- methylenedioxy) 'phenylprop2-(E)-enyl 160 3-cyclohexylpropyl 3- cyclohexylprop-2-(E)enyl (IR)-1,3-diphenyl-lpropyl 3-phenylpropyl It 3-(2,5-dimethoxy) phenylpropyl 3 - (2, 5 -dime, thoxy) phenylprop-2-(E)-enyl 450 2-(3,4,5-t.rimethoxy phenyl)ethyl 3-(3-pyridyl)propyl 3-(2-py,ridyl)propyl 3-(4-pyridyl)propyl 3-phenylpropyl 11 8 82 95 1025 1400 3 9 20 Y.I.

42 600 52 4000 92 1970 250 5 195 23 1000 36 In mammalian cells, FKBP-12 complexes with the inositol triphosphate receptor (IP3R) and the ryanodine receptor (Ry,). It is believed that the neurotrophic compounds of this invention disassociates FKBP-12 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 FKBP-12 as the IP,R receptor, one could assume that the drugs displace the channels from FKBP-12.

Chick Dorsal Root Ganglion Cultures and Neurite Outgrowth Dorsal root ganglia were dissected from chick embryos of ten day gestation. Whole ganglion explancs were cultured on thin layer Matrigel-coated 12 well plates with Liebovitz LIS plus high glucose media supplemented with 2mM glutamine and ioli fetal calf serum, and also containing 10 uM cytosine 3-D arabino-Furanoside (Ara C) at 370C in an environment containing 5g CO., twenty-four hours later, the DRGs were created with various concentrations of nerve, growth factor, immunophilin ligands or combinations c':

NFG plus drugs. Forty-eight hours after drug treatment, the ganglia were visualized under phase contrast or Hoffman Modulation contrast with a Zeiss Axiovert inverted microscope. Photomicrographs of the 37 explants were made, and neurite outgrowth was quantitated. Neurites longer than the DRG diameter were counted as positive, with total number of neurites quantitated per each experimental condition. Three to four DRGs are cultured per well, and each treatment was performed in duplicate.

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

TABLE II

Example No.

1 2 3 4 5 6 10 11 14 is 16 17 18 19 20 21 30 22 23 24 25 26 35 27 28 29 30 response curve for Neurite Outgrowth in Chick DRG ED.., neurite outgrowth, nM 53 105 149 190 10 75 0.46 0.015 2 0.8 0.015.

0.05 30 6 0.13 0.025 0.66 1100 0.014 0.50 2 500 0.50 10 100 Sciatic Nerve Axotomy Six-week old male Sprague-Dawley rats were 38 anesthetized, and the sciatic nerve exposed and crushed, at the level of the hip, by forceps. Test compounds or vehicle were administered subcutaneously 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 significant decrease in the number of axons (5011 decrease as compared to non-lesioned control) and degree of myelination (909.- decrease as compared to nonlesioned 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 is lesion, resulted in significant regeneration of both axon number (516 decrease as compared to non-lesioned control) and the degree of myelination (509c 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 butwere not lesioned; "Vehicle" denotes animals that were les ioned and received only vehicle (i.e., no drug) Example 1 showed a striking similarity to the sham treated animals, demonstrating the powerful neuroregenerative effects of these compounds in vivo.

r 39 Inactive is a compound that is inactive as an FKIBP12 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 FKBP12. Quantitation for these data are shown in Table III.

Table III

Treatment Axon Number Myelin Level (-0. Control) Sham 100 100 Lesion:

+ vehicle (s.c.) so 10 + Example 1 100 so (3 0 mg/ is kg s.c.) + Inactive 25 25 (30 mg/kg s.c.) MPTP Model of Parkinson's Disease in Mice MPTP lesioning of dopaminergic neurons in mice was used as an animal model of Parkinson'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 days, as well as for an additional S 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 [3H]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 hydoxylase 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 TH-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 C3H]-CFT binding, relative to animals receiving MPTP but not the Guilford compounds. Fig-ure 5 gives this data in bar graph form. It is shownthat 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 Exampie 17, as compared to animals that received lesioning agent but no drug (MPTP/Vehicle).

The following examples are illustrative of preferred embodiments of the invention and are not to be construed as limiting t-he invent--ion thereto. All 41 preferred embodiments of the invention and are not to be construed as limiting the invention thereto. All polymer molecular weights are mean average molecular weigh!--s. All percentages are based on the percent by weigh: of the final delivery system or formulation prepared unless otherwise indicated and all totals equal 100-0o by weight.

EXAMPLES -he inventive compounds may be prepared by a variez:y of synthetic sequences that utilize established chemical transformations. The general pathway to the present compounds is described in Scheme.1. Nglyoxylproline derivatives may be prepared by reacting L-proline methyl ester with merhyl 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 propyi, esters and amides of the invention.

42 Scheme I M. =CC:'3 a c AY CC-42 G 0.

CC-12 N - C] C3 c CC-43 AU or F;NtcX Y-Z =H3 N1e0H/H a CH N N 2 Coupling meL'icd L, Cl G (3 cl C Y-Z N 11 (2 c Z cl R EXAMPLE 1 Synthesis of 3 -phenyl 1-propyl (2S)-1-(3,3-dimethyl1,2-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 OOC and treated with triethylamine (3.92 91 - 2.1 eq). After stirring the formed slurry 38.74 mmol.

under a nitrogen atmosphere for 15 min, a solution of -methyl oxalyl chloride (3,_20 g; 26.12 mmol) in methylene chloride (45 mL) was added dropwise. The result:ing mixture was stirred at OIC for 1.5 hr. After filtering to remove solids, the organic phase was washed with water, dried over MgSO, and concentrated.

43 The crude residue was purified on a silica gel column, eluting with 501k ethyl acetate in hexane, to obtain 3.52 g (881) of the product as a reddish oil. Mixture of cis-trans amide rotamers; data for trans rotamer given. 'H NMR (CDCl.): d.1. 93 (dm, 2H); 2.17 (M, 2H) 3. 62 (m, 2H); 3. 71 (s, 3H); 3.79, 3.84 ( s, 3H total) 4.86 (dd, 1H, J = 8.4, 3.3). synthesis of methyl (2S)-1-(1,2-dioxo-3, 3dimethylpentyl) - 2 -pyrrolidinecarboxylate. 10 A solution of methyl (2S)-1-(1,2-dioxo-2methoxyethyl)-2-pyrroliclinecarboxylate (2.35 g; 10.90 mmol) in 30 mL of tetrahydrofuran (THF) was cooled to -78C and treated with 14.2 mL of a 1.0 M solution of 1,1-d,--nethylpropylma,inesium chloride in THF. After is stirring the resulting homogeneous mixture at -780C 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 (750-.) of the oxamate as a colorless oil - 'H NMR (CDC13): d 0. 88 (t, 3W; 1. 22, 1. 26 (s, 3H each); -1. 75 (dm, 2H); 1. 87-2. 10 (m, 3H); 2.23 IH); 3.54 (m, 2H); 3.76 (s, 3H); 4.52(dm, 1 J = 8. 1-, 3.4) Synthesis of (2S)-1-(1,2-diaxo-3,3dimethylpentyl) -2- pyrroli dinecarboxylic acid.

44 A mixture of methyl (2S)-1-(1,2-dioxo-3,3dime thyl pent yl) -2pyrrolidinecarboxylate (2.10 g; 8.23 mmol), 1 N LiOH (15 mL), and methanol (50 mL) was -stirred at O'C for 30 min and at room temperature overnight. The mixture was acidified to PH 1 with i i\T HCl, 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 snowwhite solid which did not require further purification.

1H MCR, (CDC13): d 0. 87 (t, 3H); 1. 22, 1. 25 (s, 3H each); 1. 77 (dm, 2W 7 2. 02 (m, 2H); 2. 17 (m, 1H); 2.25 (m, i--q); 3.53 (dd, 2H, J = 10. 4 f. 7. 3); 4. 55 (dd, 1H, J = 8. 6, 4. 1).

Synthesis of 3 -phenyl- 1-propyl (25)-1-(313- dimetnyl - 1, 2 -dioxopentyl) -2 -pyrrolidinecarboxylate (Exam-p 1 e 1). A mixture o f (2 S) - 1 - (1, 2 - dioxo - 3, 3 - dime t.,iylpentyl) -2-pyrrolidine-carboxylic acid (600 mS; 2.49 mmol), 3 -phenyl- 1-propanol (508 mg; 3.73 mmol), dicyc--iohexylcarbodiimide (822 mg; 3.98 mmol), campl-orsulphonic acid (190 mg; 0.8 mmol) and 4 dime t'-iyl aminopyridine (100 mg; 0.8 mmol) in methylene chloride (20 mL) was stirred 'overnight under a nitrogen.

atmos::)here. The reaction mixture was filtered through Celite to remove solids and c-oncentrated in vacuo, and the c-rude material was purified on a flash column (25% ethyl acetate in hexane) to obtain 720 mg (80%) of Example 1 as a colorless oil. IFI NMR (CDC'3): d 0.84 (t, 3--:-:); 1. 19 (s, 3H); 1. 23 (S, 3H); 1.70 (dm, 2H); 1. 98 (m, SH); 2. 22 (m, 1F1); 2 -64 (M, 2H); 3.47 (M, 2H) 4. 14 (m, 2H); 4. 51 (d, 1H); 7. 16 (M, 3H); 7. 2 6 (M, 214) The method of Example 1 was utilized to prepare the following illustrative examples: Example 2: 3 -phenyl - 1-prop-2 - (E) enyl (2S) -1(3,3dimethyl-1, 2-dioxopentyl) -2-pyrroliclinecarboxylate, 801k., 'H NMR (360 Mhz, WC13): d 0.86 (t, 3H); 1.21 (s, 3H) 1. 25 (s, 3H) 1. 54-2. 10 (m, SH).; 2. 10-2. 37 (m, 1H) 3. 5 2 - 3. 5 5 (m, 2H); 4. 5 6 (dd, 1H, J = 3. 8, 8. 9) 10 4.78-4.83 (m, 2H) 6.27 (M, 1H); 6.67 (dd, 1H, J 15. 9); 7. 13 - 7. 5 0 (m, SH).. Example 3: 3-(3,4,5-trimethoxyphenyl)--propyl (2S)-1(3,3-dimethyl-1, 2- dioxopentyl) -2-pyrrolidine- carboxylate, 612k, 'H NMR (CDC'3): d 0. 84 (t, 3H); 1. 15 (s, 3HT); 1. 24 (s, 3H); 1 - 71 (dm, 2H); 1. 98 (m,SH); 2. 2 4 (m, 1H) 2. 6 3 (m, 2H); 3 - 5 1 (t, 2H) 3. 7 9 (s, 3 H) 3. 8 3 (s, 3H) 4. 14 (m, 2H); 4.5 2 (m, 1H) 6. 3 6 (s, 2H) Example 4: 3-(3,4,5-trimethoxyphenyl)-1-prop-2-(E)enyl (2S)-1-(3,3dimethyl-1,2-clioxopentyl)-2-pyrrolidine carboxylate, 66-0k, 'H NMR (CDC13): d 0.85 (t, 3H); 1.22 (s, 3 H); 1. 2 5 (s, 3 H); 1. 5 0 - 2. 11 (m, SH); 2. 11 - 2. 4 0 (M, 1H); 3. 5 5 (m, 2H); 3-._8 5 (s, 3 H); 3. 8 8 (s, 6H); 4. 5 6 (dd, 11-71); 4. 8 1 (m, 2 H); 6. 2 2 (m, 1 H); 6. 5 8 (d, 1 H, J 16); 6.63 (s, 2H).

Example 5: 3-(4,5-methylenedioxyphenyl)-1-propyl (2S) 1-(3,3-dimethyl-1,2dioxopentyl)-2-pyrrolidine- 46 carboxylate,8215, '.H NMR (360 MHz, WC1,): d 0.86 (t, 3 H); 1. 2 2 (s, 3 1-1); 1. 2 5 (s, 3 H) 1 - 6 0 - 2. 10 (m, 3. 36-3. 79 (m, 2H); 4. 53 (dd, 1H, J 3.8, 8.6); 4. 614.89 (m, 21-1); 5 - 96 (s, 2H); 6710 (m, 1H); 6. 57 (dd, 1H, J = 6. 2, 15. 8); 6. 75 (d, 1H, J = 8. 0) 83 (dd, 1H, j = 1. 3, 8. 0); 6. 93 (s, 1H).

Example 6: 3 - (4, 5 -methylenedioxypheny!) - 1 -prop -2 -(E) - enyl (2S) -1- (3,3-dimethyl-1,2-dioxopentyl) -2 pyrrolidinecarboxylate,820l5, 'H NMR (360 MHz, CDC13 d 0. 86 (t, 3H); 1.22 (s, 3H); 1.25 (s, 3H); 1.602.10 (M, SH); 2.10-2.39 (m, 1H); 3.36-3.79 (m, 2H); 4.53 (dd, 1H, J 3. 8, 8. 6); 4. 61-4.89 (m, 2H); 5. 96 (s, 2H); 6. 10 1H); 6. 5 7 (dd, 1H, J = 6. 2, 15. 8); 6. 7 5 (d, 1H, J 8. 0); 6. 8 3 (dd, 1H, J = 1. 3, 8. 0); 6. 9 3 (s, is 11-1).

Example 8: 3 -cyclohexyl-l-prop-2- (E) -enyl (2S) -1- (3, 3 - dimethyl-1, 2-dioxopentyl) -2-pyrrolidinecarbecylate, 920%-, 'H NMR (360 MHz, CDC13): d 0.86 (t, 3H); 1.13-1.40 (m + 2 singlets, 9H total); 1. 50 -1. 87 (m, 8H); 1. 87 2.44 (m, 6H); 3.34-3.82 (m, 2H); 4.40-4.76 (m, 3H) 5. 35-5. 60 (M, 1H); 5. 60-5.82 (dd, 1H, i = 6. 5, 16) Example 9: (IR)-1,3-Diphenyl-l-propyl (2S)-1-(3,3 dimethyl - 1, 2 - dioxopencyl) - 2 -pyrrol idinecarboxylate, 9 O'E, '.H NMR (360 MHz, WC13-): d 0.85 (t, 3H); 1.20 (S, 3H) 1. 2 3 (S, 3H); 1. 4 9 - 2. 3 9 (m, 7F1) 2. 4 6 - 2.8 6 (m, 2H) 3.25-3.80 (m, 2R); 4.42-4.82 (m,!); 5.82 (td, 1H, J = 1. 8, 6. 7); 7. 05 -7. 21 (m, 3H) 7.21-7. 46 (m, 7H).

47 Example 10: 3 -phenyl -1 -propyl (2S)-1-(1,2-diaxo-2-E2 furanyll)ethyl -2 -pyrroliclinecarboxylate, 990-, 'H NMR (300 MHz, WC1,): d 1.66-2.41 (m, 6H); 2.72 (t, 2H, i 7. 5); 3. 75 (m, 2H); 4.21 (m, 2H); 4. 61 (m, 1H); 6. 58 (m, 1 H); 7. 16 - 7. 2 9 (m, 5 H); 7. 7 3 (M, 2 H).

Example II: 3-phenyl-i-propyl (2S)-1-(1,2-dioxo-2-[2 thienyll)ethyl-2-pyrrolidinecarboxylate, 8look, 'H NMR (300 MHz, WC1,): d 1.88-2.41 (m, 6H); 2.72 (dm, 2H); 3. 7 2 (m, 2H); 4. 0 5 (M,!H); 4. 2 2 (M, 1H); 4. 64 (m, 1H) 7. 13 - 7. 2 9 (m, 6H); 7. 7 5 (dm, 1W; 8. 0 5 (m, IH).

Example 13: 3 -phenyl - 1 -propyl C2S) - 1 - (1, 2 -clioxo- 2 phenyl)ethyl-2-pyrralidinecarboxylate, 9915, 'H NMR (300 MHz, CDC13): d 1. 9 7 - 2. 3 2 (m, 6H) 2. 74 (t, 2H, J = 7. 5); 3. 5 7 (m, 2H); 4. 2 4 (M, 2 H) 4. 6 7 (m, 1H); 6. 9 5 - is 7. 2 8 (m, SH); 7. 5 1 - 7. 6 4 (m, 3H); 8. 0 3 - 8. 0 9 (m, 2H) - Example 14: 3-(2,5-dimethoxyphenyl)-1-propyl (2S) -1 (3, 3 -dimethyl - 1, 2 -dioxopentyl) - 2 -pyrrolidine - carboxylate, 990-., 'H NMR (300 MHz, CDC13): d 0.87 (t, 3H) 1 - 22 (s, 3H); 1. 26 (s, 3H) 1. 69 (m, 2F1) 1 96 SH) 2.24 (m, 1H); 2. 68 (m, 2H) 3. 55 (m, 2H) j 75 3H) 3.77 (s, 3H); 4. 17 (M, 2H) 4.53 (d, 1H) 6. 72 3H).

Example 15: 3-(2,5-dimethoxyphenyl)-1-prop-2-(E)-enyl (2S) -1(3,3-dimethyl-l, 2-di.,oxopentyl) -2pyrrolidine carboxylate, 99%, 'H NivIR (300 MPz, CDC13): d 0.87 (t, 3H) 1. 22 (s, 3W 1 - 26 (s, 3H) 1. 67 (m, 2H) 1. 78 (m, 1H) 2.07 (m, 2H) 2.26 (m, 1H) 3.52 (m, 2H) 3. 78 (s, 3H) 3. 80 (s, 3H) 4 -54 (rn, 1H) 4. 81 (m, 2H) 6.29 (m, (S, (M, 48 (dt, 1H, J = 15.9); 6.98 (5, 1H). Example 16: 2 - (3, 4, 5 - trimethoxyphenyl) -1 -ethyl (25)-1(3,3-dimethyl-1,2-dioxopentyl)-2- pyrrolidinecarboxylate, 97%, 'H NMR (300 MHz, CDC13): Cl 0. 84 (t, 3H); 1. 15 (s, 3H); 1 - 24 (s, 3H); 1. 71 (dm, 2H); 1. 9 8 (m, SH); 2.24 (m, 1H); 2.63 (m, 2H); 3.51 (C, 2H) 3.-79 (s, 3H); 3.83 (s, 3H); 4. 14 (m, 2H); 4. 52 (m, 1H) 6. 3 6 (s,2H). Example 17: 3-(3-Pyridyl)-1-propyl (2S)-1-(3, 310 dimethyl-l, 2-dioxopentyl) -2-pyrrolidinecarboxylate, 80-., 'H INMR (CDC13, 300 MHz): d 0. 85 (t, 3H); 1. 23, 1. 26 (s, 3H each); 1. 63 -1.89 (m, 2H) 1. 90-2.30 (m, 4H); 2.30-2.5.0 (m, 1FI); 2.72 (t, 2H) 3.53 (M, 2H); 4.19 (m, 2H); 4.53 (m, 1H); 7.22 (m, 1H); 7.53 (dd, is 1I-1); 8.45.

Example 18: 3-(2-Pyridyl)-1-propyl (2S)-1-(3,3dimethyl-l, 2-dioxopentyl) 2-pyrrolidinecarboxylate, 882k, 'H NMR (CDC13, 300 MHz): d 0.84 (t, 3H); 1.22, 1. 27 (s, 39 each); 1. 68-2.32 (m, 8H); 2.88 (t, 2H, J 7. 5); 3. 5 2 (m, 2 H); 4. 2 0 (m, 2H); 4. 5 1 (m, 1H); 7. 0 9 7. 19 (m, 2 9); 7. 5 9 (m, 1H); 8. 5 3 (d, 1H, J = 4. 9) -Example 19: 3-(4-Pyridyl)--1-propyl (2S)-1-(3,3dimethyl-l, 2-dioxopentyl) -2-pyrrolidirlecarboxylatz-, 910k, 'H NMR (CDC13, 300 MHz): d 6-92-6.80 (M, 4H) l 6.28 (m, IF1); 5. 2 5 (d, 1H, J = 5. 7); 4. 12 (m,!H). 0 8 (S, 3H); 3.79 (s, 3H) 1- 3.30 (m, 2H); 2. 33 (m, IH) 1.851. 22 (m, 7H); 1. 25 (s, 3H); 1. 23 (s, 3H); 0. 89 (r-, 3H, J = 7. 5) - 49 Example 20: 3 -phenyl -1-propyl (25)-1-(2-cyclohexyl-1,2dioxoethyl)-2- pyrralidinecarboxyla--e, 910-.,!H NMR (CDC131 300 MHz): d 1.09-1.33 (m, SH); 1.62-2.33 (m, 12H); 2.69) t, 2H, J = 7. 5); 3. 15 (dm, IH); 3 - 68 (m, 2H) 4.16 (m, 211) 4.53, 4.84 (d, 1H total); 7.19 (m, 3H) 7.29 (m, 2H) Example 21: 3 -phenyl - 1 -propyl (2S) - 1 - (2 - tert- butyl - 1, 2 dioxoethyl)-2-pyrrolidinecarboxylate, 920k, 'H NMR (CDC13o 300 MHz): d 1.29 (s, 9H); 1.94-2.03 (m, SH) 2.21 (m, 1H); 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-cyclohexylethyl-!, 2-dioxoethyl) -2- pyrrolidinecarboxylate, 971k, 'H NMR (CDC1,, 300 MHz): d 0.88 (m, 2H); 1. 16 (M, 4H); 1.43-1.51 (m, 2H); 1.67 (m, SH); 1.94-2.01 (m, 6H); 2.66-2.87 (m, 4.86 (m, 1H); 7.17-7.32 (m, SH 4H) 3 62-3 77 (m 2H); 4.15 (m, 2H); Example 23: 3- (3-pyridyl) -1-propyl (2S) -1- (2-eyclo hexylethyl - 1, 2 dioxoethyl) -2 -pyrrolidinecarboxylate, 7 0 C5, 'H NMR (CDC'3, 300 MHz): d 0.87 (m, 2H) 1.16 (m, 4H); 1. 4 9 (m, 2H); 1. 6 8 (M, 4H); 1. 9 5 - 2. 3 2 (m, 7H); 2. 71 (m, 2H); 2. 85 (m, 2H); 3. 63 - 3.78 (m, 2H) 4. 19 (m, 2H); 5.30 (m, IH); 7.23 (m, 1H); 7.53 (m, IH) 8.46 (m, 2H) Example 24: 3- (3-pyridyl) -1-propyl (2S) -1- (2- terC butyl-1,2-dioxoethyl)-2-pyrrolidinecarboxylate, 8355, 'H NMR (CDC'3, 300 MHz): d 1.29 (s, 9H); 1.95-2.04 (m, SH); 2.31 (m, 1F0; 2.72 (t, 2H, J = 7. 5); 3.52 (m, 2H) so 4. 18 (m, 2H); 4. 5 2 (m, 1H); 7. 19 - 7. 2 5 (M, 1I-1); 7. 5 3 (m, 1H); 8.46 (m., 2H).

Example 25: 3,3-diphenyl-l-propyl (2S)-1-(3,3-dimethyl1,2-dioxopentyl)-2pyrrolidinecarboxylate, 99%, 'H NMR (CDC'3, 300 MHz): d 0.85 (t, 3H); 1. 21, 1.26 (S, 3H ea.ch); 1.68-2.04 (m, SH); 2.31 (m, 1H); 2.40 (m, 2H) 3. 5 1 (m, 2H); 4. 0 8 (m, 3 H); 4. 5 2 (m, 1FI); 7. 18 - 7. 3 1 lOH) Example 26: 3- (3-pyridyl) -1-propyl (2S) -I- (2-cyclohexyl-1,2-dioxoethyl)-2-pyrroli-dirlecarboxylate, 885k, 'H NMR (CDC'3, 300 MHz): d 1.24-1.28 (m, SH); 1.88-2.35 1 1H); 2. 7 2 (t, 2H, J = 7. 5); 3 0 0 - 3. 3 3 (dm, IH) (m, 2H); 4.19 (.m, 2H);.55 (m, 1H); 7.20-7.24 1H); 7.53 (m, 1H); 8.47 (m, 2P1). Example 27: 3- (3-Pyridyl) -1- propyl (2S) -N- ( [2-thienyll glyoxyl)pyrrolidinecarboxylate, 49-0.-, 'H NMR (CDC'3, 300 MHz): d 1.81-2.39 (m, 6H); 2.72 (dm, 2H); 3.73 (m, 2H); 4. 2 1 (m, 2 H) 4. 9 5 (m, 1 H) 7. 19 (m, 2 H) 7. 6 1 (M, 1H) 7 - 8 0 (d, 1H) 8. 0 4 (d, 1H) 8. 4 6 (m, 2 H) Example 2 8: 3, 3 -Diphenyl - 1 - propyl (2 S) - 1 - (3, 3 - dimethyl 1, 2-dioxobutyl) -2- pyrrolidinecarboxylate, 99%, 'H NMR (CDC13. 3oo MHz). d 1. 27 (s, 9H); 1. 96 (rn, 2W; 2.44 (m, 4W; 3.49 (m, IH); 3.64 (rn, IH); 4.08 (m, 4W'; 4.53 (dd, 1H); 7. 24 (m, lOH) Example 29: 3,3-Diphenyl-l-proov! (2S) -1- cyclohexyl glyoxyl-2-pyrrolidinecarboxylae, 91%, 'H NIMR (CDC131 300 mHz): d 1. 32 (m, 6H); 1. 54 -2. 41 (m, lOH); 3. 20 (dm, IH); 3. 6 9 (m, 2H); 4.12 (m, 4H); 4. 5 2 (d, 1H) 3.69 51 7.28 (m, lOH). Example 30: 3,3-Diphenyl-1-propyl (2S) -1- (2-thienyl) glyoxyl-2-pyrrolidinecarboxylate, 75051;, 'H NMR (WC13 300 MHz): d 2.04 (m, 3H); 2.26 (M, 2H); 2.48 (m, 1H 3. 70 (m, 2H); 3.82 -4. 18 (m, 3H total); 4. 64 (m, JH) 7. 2 5 (m, 1 1H); 7. 7 6 (dd, 1H); 8. 0 3 (m, IH).

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 crans-cinnamates; these latter may be reduced to the saturated alcohols by reaction with is 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 Crans-cinnamates may be reduced to (E)-allylic alcohols by the use of dilsobutylaluminum hydride.

Lichium aluminum R-CHO Ph 3P-CHCOOCH3 R CCOCH,, hydride THF H 2.

Drisabuoaiuminum Pd/C hydride v Scheme II 2 COOCH:3 Lithium aluminum hydride or Diisabucylaluminum hydride ::: 2 Longer chain alcohols may be prepared by homologat-ion of benzyl-ic and nigher aldehydes. Alternatively, these aidenvaes may be prepared by conversion of the corres-ocn-'Ing 5 acids, and ch-enethyl and Genera- zrccedure fcr F o r 7, e -v' esters, exemp-if ed trans - c i nna-,-,ia t e A sc--',-zi--n of ',4,5-z:r--,-,ez:hcxvbenzaidehvde (5.0 q- 2 5. 4 8 mmc-',, and m, e t hy 1,'zi r e Jdene)ace7_a--e g; 29.9i mmol) in phosphorany tetrahydrofuran (250 mL) was refluxed overnight. Af:zer cooling, tne- reaction mixtu_-_ was diluted wi7-h 200 m_7 c-f et-v-- and wased dried, and --c-cen:=ed in va-cuo. The crude residue was c.iromatogra-zned on a silica ge-I cclumn, elut-ing with 2506 ethyl acezate in hexane, --o ciDtain 5.63 cr (881) of the cinnamaze as a white crystalline solid, Mhz; CDCI,) d 3.78 (s, 3H) 13.85 (s, 6-H) 6.32 (d, 1H, J = 16) 6.72 (s, 2H); 7_559 (d, 1-H, i 16) General orocedure for --he sy-nthesis of saturated alcohols from acrylic esters. Exemplified for (3,4,5trimethoxy) phenylpropancl.

A soluzion of methyl (3,3,5-trimethoxy) -Cranscinnamate (1.81 g; 7.17 mmcl.) in tetra-hydrofuran (30 mL) was added -in a dropwise manner to a solution of lithium aluminum hydride (14 mmol) in THF (35 m.L), with stirring and under an argon atmosphere- After the p-henylacetic and a"cciols.

e s i s c -f a c ry -i Ii c 5 - t: r i me 7_ hoxy) - is 2 x 200 -,j- c-: water, 1 H NMR (3 C Cj is 53 addition was complete, the mixture was heated to 75'C or 4 hours. Af ter cooling, J t- was quenched by the careful addition of 15 mL of' 2-,\T NaOH followed by 50 m7, of water. The resulting mixture was filtered through Celite to remove solids, and chee -filter cake was washed fractions were th ethyl ace, -=:e. The combi-ed organic washed with water, dried, concennrated in vacue, and purified on a silica gel column, eluting with ethyl acetate to obtain 0.86 g (53%) c.-: the alcohol as a clear cl, '-K _W,.R El 8 7 (m, 2 g.) 1 2. 6 (s, 3H); 3.83 (s, 6H1); 6.40 (s, 2F1).

General procedure for the synthesis of Crans- ally-lic alcoholls from acryl-,c es-,-ers. Exemplified for oxy) phenylprop - 2 -, E) - enol - - - A solut ion of methyl (3, 3, 5 - tr-ime-thoxy) - cranscinnamate (1.35; 5. 35 mmc-") in toluene (25 rnL) was cooled to -101C and treated with a solution off diisobutylaluminum hydride in ioli-.,ene (11.25 m-7 of a 1.0 M solution; 11.25 mmol). The reaction mixture was stirred for 3 hrs at O'C and then quenched with 3 mL otc methanol followed by 1 N 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 -n hexane furnished 0.96 g (80%) of a thick oil, -H NMR (360 Mhz; CDCl.): d 3.85 (s, 3H); 3.87 (s, 61-1); 4.32 (d, 2H, i = 5 - 6); 6.29 (300 Mhz; CDC13): d 1. 23 (br, 11H); (t:, 21.1, J = -7. 1); 3. 6 6 (L, 2H); 3. 8 0 54 (dt, 1H, J = 15. 8, 5. 7), 6. 54 (ci, 1H, J = 15.8); 6 -61 (5, 2W.

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

Such f rom

Claims (4)

CLAIMS: 1. A chemical compound comprising (2S)-1-(1,2-dioxo-3,3dimethylpentyl)-2pyrrolidinecarboxylic acid. 2. A method of svnthesizina a chemical compound according to claim 1, comprising: forming a mixture of methyl (2S)-1-(1,2dioxo-3,3-dimethylpentyl)-2-pyrrolidine- carboxylate, 1N lithium hydroxide, and methanol; stirring the mixture at OOC for 30 minutes and at room temperature for about 12 hours; acidifying with 1N hydrochloric acid to pH 1; diluting with water; and 10 extracting the organic phase into methylene chloride. is 56 3. A neurctrophic compound of the formula: X is _Z 0 R, is a C.-C, scraight: or branched chain or alkenv! cTrou:) ontionally substizuted with C3-CS cycloalkyl, C3 or C. cycloalkyl, C,-C, cycloal'.v,er-yl, or Arl, where said alkyll, alkenyl, cycloalkyl or cycloalkeny! groups may be optionally substituted with C,-C, alkyl, C,-C, alkenyl, or hydroxy, and where Ar, is selected from the group consisting of I-napthyl, 2-napthyl, 2-indolyl, 3-indolyl, 2-furyl, 3-fury 1, 2-thienyl, 3-thienyl, 2-, 3-, or 4-pyridyl, or phenyl, having one to three substituents which are independently selected from the group consisting of hydrogen, halo, hydroxyl, nitro, trifluoromethyl, C,-C, straight or branched Y is - 20 z z 5alkyl or alkenyl, C,-C, alkoxy or C,-C, alkenyloxy, phenoxy, benzyloxy, and amino; is oxygen, sulfur, methylene (CH2), or 112; is oxygen or NR2, where R2 is hydrogen or C,C. alkyl; and is a C2-C6 straight or branched chain alkyl or alkenyl, wherein the alkyl chain is substiuted in one or more positions with.Ar, as defined above, C3-C, cycloalkyl', cycloalkyl connected by a C,-C6 straight or unbranched alkyl or alkenyl chain, or Ar. where Ar2 is selected from the group consisting of 2--indolyl,3-indolyl, 2furyl, 3-furyl, 2--Lh-iazolyl, 2-thienyl, 3thienyl, 2-, 3-., or 4- pyridyl, or phenyl, having one to three substituents which are independently selected from the group consisting of hydrogen, halo, hydroxyl, nitro, trifluoromethyl, C,-C, straight or branched alkyl or alkenyl, C,-C, alkoxy or C,C4 alkenyloxy, phenoxy, benzyloxy, and amino; may also be the fragment: 0 - CH 11 X-R R 3 4 where R3 is selected from the group consisiting of straight or branched alkyl Cl-Cs optionally substituted with C3-Cl cycloalkyl, or Ar, as defined above; X2 is 0 or -NTR,, where R. is selected from the group consisting of hydrogen, Cl-C6 straight or branched alkyl and alkenyl; R4 is selected from the group consisting of phenyl, benzyi, Cl-C, s- raight or branched alkyl or alkenyl, and C,-C, straight or branched alkyl or alkenyl substituted with phenyl; or pharmaceutically acceptable salts or hydrates Chereof_. +. The neurocrophic compound of claim 5, which has an affinity for FK1P3Ptype immunophilins. 5, The neurotrophic compound of claim 41, where the FKBP-type immunophilin is FKBP-12. 16. The neurotrophic compound of claim 3L, capable of inhibiting rotamase activity. The neurotrophic compound of claim3, where Z and R, are lipophilic groups. sq 19. The neurotrophic compound according to claim 3. that is selected from the group consisting o= j- 3 -phenyl -1 -propyl (2S)-1-(3,3-dimethyl-1,2dioxopentyl) -2-pyrro- lidinecarboxylar-e-, 3 -phenyl -1-prop-2- (E) -enil, (2S'-'-'-(3,3d-iL-nethyl1, 2 - dioxopentyl) - 2 -.oyrrol idinecarboxylate, 3 - (3, 4, 5 - t rime c hoxypheny!) - 1 -propyl (25) 1 - (3, 3 dimethyl - 1, 2 -dioxopentyl) - 2 -pyrrol idinecarboxylate, 3-(3,4,5-trimethoxyphenyl)-1-r)rop-2-(E)-eny1 (2S)1-(3,3-dimethyl-1,2-dioxoDenty'-.)-2- pyrrolidinecarboxylate, 3-(4,5-dichlorophenyl)-1-r)ropyl (2S)-1-(3,3dimethyl-1, 2-dioxoperityll) 2-py----olidinecarboxylate, 3-(4,5-dichloropherivl-)-!-,jror,-2-(-:')-enyl (2S) -1- 1 - (3, 3 dime thyl - 1, 2 -dioxoiDentyl) -2pyrrolidinecarboxylate, 3-(4,5-methylenedioxyphenyl)-1-propyl (2S)-1-(3,3dimethyl-l, 2- dioxopentyl) -2-pyrrolidinecarboxylate, 3 - (4, 5 -methylenedioxyphenyl) - 1 -prop - 2 - (E) - enyl (2 S) - 1 (3, 3 - dime thyl - 1, 2 - dioxopentyl) - 2 pyrrolidinecarboxylate, 3-cyclohexyl-l-propiyl (2S)-1-(3,3-dimethyl-1,2dioxopentyl) -2pyrrolidinecarboxylate, 3-cyclohexyl-l-prop-2-(E)-enyl (2S)-1-(3,3dimethyl-1, 2-dioxopentyl) -2-pyrrolidinecarboxylate, (1R)-1,3-diphenyl-l-propyl (2S)-1-(3,3-dimethyl1, 2-dioxopentyl) -2pyrrolidinecarboxylate, (1R)-1,3-diphenyl-l-prop-2-(E)-eny1 (2S)-1-(3,3dime rhyl-1,2-dioxopentyl) -2-pyrrolidinecarboxylate, (IR) -1-cyclohexyl-3- phenyl -l -propyl (2S) -l- (3,3dimethyl- l, 2dioxopentyl) -2-pyrrolidinecarboxylate, (1R) -l -cyclohexyl-3 -phenyl- l -prop -2- (E) -enyl (25) 1 (3, 3- dime thyl- l, 2-dioxopentyl) -2pyrrolidinecarboxylate, (1R) -l- (4, 5-dichlorophenyl) - 3 -phenyl -1 -propyl (2S) -l- (3, 3- dimethyl-l, 2-dioxopentyl) -2pyrrolidinecarboxylate, 3-phenyl-l-propyl (25)-1-(1,2-dioxo-2cyclohexyl) ethyl -2 -pyrrolidinecarboxylate, 3 -phenyl- 1-propyl (25)-1-(1,2-dioxo-4cyc 1 ohexyl) butyl - 2 - pyrro 1 idinecarboxyl ate, 3 -phenyl - 1-propyl (25)-!(!,2-dioxo-2-[2f uranyll) ethyl -2pyrrolidinecarboxylate, 3 -phenyl -1-propyl (2S)-1-(1,2-dioxo-2-[2thienyll)ethyl -2- pyrrolidinecarboxylate, 3-phenyl-l-propyl (2S)-1-(1,2-dioxo-2-E2thiazolyll)ethyl -2 pyrrolidinecarboxylate, 3 -phenyl- 1 -propyl (2S)-1-(1,2-dioxo-2 phenyl) ethyl -2 - pyrrolidinecarboxylate, 1,7-diphenyl-4-heptyl (2S)-1-(3,3-dimethyl-1,2dioxopentyl) -2- pyrrolidinecarboxylate, 3 -Phenyl -1-propyl (2S)-1-(3,3-dimethyl-1,2-dic)xo- 4-hydroxybutyl) -2-pyrroli.dinecarboxylate, 3 -phenyl -1-propyl (2S)-1-(3,3-dimethyl-1,2dioxopentyl) -2- pyrrolidinecarboxamide, l- [l(3,3dimethyl-1,2-dioxopentyl) -L-prolinel-7L 6# phenylalanine ethyl ester, 1-[1-(3,3 -dime t:hyl- l, 2-dioxopentyl) -L -p roline l -L - leucine ethyl ester, l- E l - (3,3-di me r-hyl- l, 2-d-ioxoS)e,-1-t-yl) -L -pro line l -L - pheny1glycine ethyl ester, l- [l- (3,3-dimethyll, 2-dioxopentyl) -Lprolinel -L- phenylalanine phenyl ester, l- [l- (3, 3 -dimethyl - 1, 2 -dioxopenr-yl) -L-praline] -L- phenylalanine benzyl ester, and l- [l- (3, 3 -dimethyl - 1, 2 -d-ioxope-nr-yl) -L-prolinel -L- isoleucine ethyl ester. v. A pharmaceutical composicion comprising a neurotrophically effeccive amcun,-t of the compound of claim3L and a pharmaceutically acceptable carrier. is IG. A method of stimulating growth of damaged peripheral nerves, which comprises: administering to damaged peripheral nerves the neurotrophic compound of claim 1 in sufficient. amounts to stimulate the growth of said nerves. it - A neurotrophic compound of the formula: 0-Z o Y R 1 0 where is z 62 is a C,-C,, su--a-igh- or bra-nched alkyl or alkenyl group optionally subs t_ ii t--ut:ed with C,-C, cvclc)a".-,vl, C, cr C,, cvcloa"tk-yr, C _C7 c y cl o a lk an y I or re s a id al k v 1, a-^fKe.-iy-',, cycloalkyl or c-cloalken-v-l croups may be oQ-C-1onally subs'--itu--ed witi. C.-C, alkyl, C,-C, alkenyl, or hydroxy, and where Ar, is selected from the croup consisting of l-napt:h., /-', 2-naothy--;, 2-indclvl, 2-furyl, 3furyl, 2-thiazolyl, 2-th-Jenyl, 3- thienyl, 2-, 3-, or i-pyridyl, or phenyl, having one to three subst:ituents which are i-,dependen--1,-Y- selecied "rem the g- -ouu consist-ing of hydrogen, '-,-alo, hydroxyl, nitro, riflucromethyl, C, -C,, straighc or branched alkyl or alkenvi, ClC, alkoxy or C,C4 alkenyloxy, phenoxy, benzyloxy, and amino; is a C2 _C6 straight or branched chain alkyl or alkenyl, wherein the alkyl chain is substituted in one or more positions with Ar, as defined above, C3C, cycloalkyl, cycloalkyl connected by a C,-C, straight' or unbranched alkyl or alkenyl chain, or Ar2 where Ar2 is selected from the group consisc-ing o-,, 2- indolvi, 3-indolyl, 2furyl, 3-furyl, 2-thiazolyl, 2-thienyl, 3thienyl, 2-, 3-, or 4-pyridyl, or phenyl, 6.3 having one to three substituents which are independently selected from the group consisting of hydrogen, halo, hydroxyl, nitro, trifluoromethyl, C,-C7. straight or branched alkyl or alkenyl, Cl-C, alkoxy or C,C4 alkeny.loxy, phenoxy, benzyloxy, and amino;_ or pharmaceutically acceptable salts or hydrates thereof. 11a. The neurotrophic compound of claim j#, wherein R, is selected from the group consisting of cl-Cg straight or branched chain alkyl, 2-cyclohexyl, 4-cyclohexyl, 2furanyl, 2- th-Jenyl., 2-thiazolyl, and 4-hydroxybutyl. 11. The neurocroph-ic compound of claim JP having an affinity for FKBPtype immunophilins. 1J. The neurotrophic compounds of claim 13, where the FKBPtype immunophilin is FKBP-12. 157. The neurotrophic compound of claim V, capable of inhibiting rotamase activity. li. The neurotrophic compound of claim, where Z and 20 R3. are liphophilic groups. 1?:. A pharmaceutical composition comprising a neurotrophically effective amount of.the compound of 64 claim 0 and a phar-maceutically accepcable carrier. is. A neurotrophic compound of the fonnula: H C N-Z N 1 0 0 where z is the fragment: 0 -CH 1 -R X2 4 3 6!; where R3 is selected from the group consisting of straight or branched alkyl C,- Cs optionally substituted with C3C, cycloalkyl, or Ar, as defined above, and unsubstituted Arl; is 0 or iYR., where R, is selected from the group consisting of hydrogen, C,-C6 straight or branched alkyl and alkenyl; is selected from the group consisting of phenyl, benzyl, C.-C, straight or branched alkyl or alkenyl, and C,-C, straight or branched alkyl or alkenyl substituted with phenyl,- or pharmaceutically acceptable salts or hydrates thereof. 117. The neurotrophic compound of claim 19, having an affinity for FKBPtype immunophilins. 110. The neurotraphic compound of claim ict, where the FKBP-type immunophilin is FKBP-12. LL The neurotrophic compound of claim le, capable of inhibiting rotamase activity. 211. The neurotrophic compound of claim 18, where Z' i a lipophilic group.

1 'i 66 2,1. A pharmaceutical composition comprising a neuror-rophically effective amount of the compound of claim 19 and a pharmaceutically acceptable carrier.

2#-. A neurotrophic compound having an affinity for FYMP-type immunophilins wherein the immunaphilin exhibits rotamase activity and the neurotrophic compound inhibits the rotamase activity of the immunophilin.

2 11. The neurotrophic compound of claim 2Y, wherein the 10 FKBP-type immunophilin is FKBP-12.

24. A method of -reaing a neurological disorder in an animal comprising administering a therapeutically effective amount oic a compound having an affinity for FKBP-type immunophilins wherein the immunophilin is exhibits rotamase activity and the neurotrophic compound inhibits the rotamase activity of the immunophilin.

2f. The method of claim 26, wherein the FKBP-type immunophilin is FKBP-12.

246 - The method of claim 20, wherein the neurological disorder is selected from the group consisting of peripheral neuropathies, and neurological pathologies related to neurodegenerati6n.

I-f 21T. The method of claim 24, wherein the neurological disorder is Alzheimer's disease.

340. The method of claim 26, wherein 'the neurological disorder is Parkinson's disease.

31. The method of claim 246, wherein the neurological disorder is amyatrophic lateral sclerosis.

3R. A method of promoting neuronal regeneration and growth in mammals, comprising administering to a subject an effective amount of a neurotrophic compound having an affinity for FKBP-type immunophilins wherein the immunophilin exhibits rotamase activity and the neurotrophic compound inhibits the rotamase activity of the immunophilin.

33. The method of claim 32, wherein the FKBP-type is immunophilin is FKBP-12.

3V. A method of preventing neurodegeneration in an animal comprising administering an effective amount of a compound having an affinity for FKBP-type immunophilins wherein the immunophilin exhibits rotamase activity and the neurotrophic compound inhibits the rotamase activity of the immunaphilin.

3.!r. The.method of claim 3.1, wherein the FKBP-type immunoz)-iiin is F1KEP-12.

36. A neurctrochic N,-cr"lyc)xyl proly! ester ccm-coun the -formula:

0-Z N ll 0 R 1 is where z cl of is a C-C-- straight or bra:,c'.-ied chain alkyl or alkenyl group CC-L- Jonally substituted with C3 to C, cycloalkyl, or A--,, where Ar, is selected from the group consisting of 2furyl, 2thienyl, or phenyl, is selected from the group consisting of oxygen and sulfur; Y is oxygen; and. 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-C6 cycloalkyl, Ar2 where A-r2 'S 2-, 3-, is 69 selected from the group consisting of or 4-nyr-idyl, or phenyl, having one to three substituents which are independently selected from the group consisting of hydrogen and C,-C, alkoxy.

3;F. The neurotrophic N-glyoxyl prolyl ester Compound of claim 34, where Z and A, are lipophilic groups.

39. The neurotrophic N-glyoxyl prolyl ester compound according to claim 34 that is selected from the group 10 consisting of:

3-.(2,5-dimethoxyphenyl)-1-propyl. (2S)-1-(3,3dimethyl-1,2-d-ioxoDentyi.) -2-pyrrolidinecarboxylate, 3- (2, 5-d-ime-(-hoxypheny!) -1-prop-2- (g) -enyl (2S) -I(3,3-dimethyl---, , 2-dioxope.nt:yl) -2-pyrrolidinecarboxylate, 2-(3,4,5-trimechoMhenyl)-1-ethyl (2S)1-(3,3dime thyl - 1, 2 - dioxopentyl) - 2 -pyrrol idinecarboxyl ate, 3-(3-Pyridyl)-1-propyl (2S)-1-(3,3-dimethyl-1,2 dioxopentyl)-2pyrrolidinecarboxylate, 3(2Pyridyl)-1-propyl (2S)-1-(3,3-dimethyl-1,2 dioxopentyl)-2- pyrrolidinecarboxylate, 3-(4-Pyridyl)-1-propyl (2S)-1-(3,3-dimethyl-1,2 dioxopentzyl)- 2pyrrolidinecarboxylate, 3 -phenyl -1-propyl (2S)-1-(2-t:ert-butyl-1,225 dioxoethyl)-2pyrrolidinecarboxylate, 3 -phenyl- 1-propyl (2S)-1-(2-cyclohexylethyl-1,2 is -qo dioxoethyl)-2-pyr--olidinecarboxylace, 3- (3pyridyl) -1-propyl (2S) -1- (2-cyclohexyler-hyl1,2-dioxoethyl)-2- pyrroldinecarboxylat--e, 3(3-pyridy!) -1-proov! (2S) 1- (2-cer:-buyl- l, 2dioxoetil-yl) -2-7--,, rrrolid-inecarboxylate, (2S)-!-(3,3-ditm.e-hvl-1,2dioxopentyl)-2-Qyrrol-idi----ecarboxylate, 3-(3.-pyridyl)-1-propyl (2S)-1-(2-cyclohexyl-1,2dioxoethyl)-2pyrrolidinecarboxylate, 3 (3-Pyridyl) -!-Dropl/1 (2S) -N- ( [2thier.ylj glyoxyl)pyrrolidinecarboxylate, 3,3-Diphenyl-l-propy! (2S)-1-(3,3-dimethyl-1,2dioxobutyl)-2-pyrrolid4- necarboxylate, 2-pyrrolid-Jnecarboxylat-.e, and 3,3-Diphenyl-1-propyl (25) -1(2-t:hienvl)glyoxy-l 2- pyrrolidinecarboxylate.

317. A pharmaceutical composition comprising a neurotrophically effective amount of the N-glyoxyl prolyl ester compound of claim 36 and a pharmaceutically acceptable carrier.

damaged jo. A method of srimulating growth oL Peripheral nerves, WhIch Com-prises:

administering to damaged peripheral nerves the neurotraphic N-glyoxyl prolyl ester compound of claim 34 in suffIcient amounts to stimula.te.-the..;growth of 71 said nerves.

44. A method of treating a neurological disorder selected from the group consisti-ng of peripheral neuropathies, and neurologi-cal pal-hologies related to neurodegeneration in an animal which comprises administering a therapeutically effective amount of a neurotrophic N-glyaxyl prolyl ester compound having an affinity for FKBP-type immunophilins wherein the immunophilin exhibits rotamase activity and the neurotrophic N-glyoxyl proly! ester compound inhibits the rotamase activity of'the immunophilin.

40- The method of claim 41, wherein the FKBP-type immunophilin is FKBP-12.

43. The method of claim 4j, wherein the neurological disorder is Alzheimer's disease.

0. The method of claim J, wherein the neurological disorder is Parkinson's disease.

4. The method of claim 4#, wherein the neurological disorder is amyotrophic lateral sclerosis.