Small molecule inhibitors of rotamase enzyme activity

Related application

This application is in part part of U.S. Patent Application Ser. No. No. 08/479436, filed May 1995. June Dec. 7, cont.

Origin of the Invention

1, Field of the Invention

The present invention relates to neurotrophic compounds having affinity for FKBP-type immunophilins, their preparation and their use for inhibiting the activity of this enzyme in relation to immunophilin proteins, in particular the activity of peptidyl-prolyl isomerase or rotamase enzymes.

2. Description of prior art

The term "immunophilin" includes several proteins that serve as receptors for the major immunosuppressive drugs, cyclosporine (CsA), FK506, and rapamycin. Known classes of immunophilins include cyclophilins and FK506 binding proteins such as FKBP. Giclosporin A binds to cyclophilin and FK506 and rapamycin bind to FKBP. These immunophilin-drug complexes are involved in many intracellular signaling systems, particularly in the immune system and nervous system.

Immunophilins are known to have peptidyl-prolyl-isomerase (PPIase) or rotamase enzymatic activity. Rotamase activity has been found to play a role in the catalysis of the transition and trans isomerization of immunophilin proteins.

Immunophilins were first discovered and studied in immune tissue. Those skilled in the art initially hypothesized that inhibition of the rotamase activity of immunophilins would result in inhibition of T-cell proliferation, thereby inducing immunosuppression with immunosuppressants such as cyclosporine, FK506 and rapamycin. Further studies have shown that inhibition of rotamase activity (alone) is not sufficient for the substance to exhibit immunosuppressive activity (Schreiber et al., Science, 1990, vol. 250, pp. 556-559). The mode of action of the drug has been shown to be the interaction of the immunophilin-drug complex with ternary protein targets (Schreiber et al., Cell, 1991, vol. 66, pp. 807-815). In the case of FKBP-FK506 and FKBP-CsA, the drug-immunophilin complexes bind to the enzyme calcineurin, an inhibitory T-cell receptor that transmits a signal that triggers T-cell proliferation. In a similar fashion, the rapamycin-FKBP complex interacts with the RAFT / FRAP protein and inhibits IL-2 receptor signaling.

High concentrations of immunophilins have been found to be present in the central nervous system. The central nervous system contains 10 to 50 times more immunophilins than the immune system. In neural tissues, immunophilins appear to play a role in nerve extraction, nitric oxide (NO) synthesis, and disconnection of the nerve impulse transducer.

Picomolar concentrations of an immunosuppressant such as FK506 and rapamictone have been found to stimulate axonal growth in PC12 cells and sensory nerves, namely spinal cord nerve cells (DRG) (Lyons et al., Proc. Natl. Acad. Sci. 1994). , vol. 91, pp. 3191-3195). In animal studies, FK506 has been shown to stimulate nerve regeneration following lesion of the vagus nerve, while animals with sciatic nerve damage restore function.

Unexpectedly, drugs with high affinity for FKBP have been found to be potent inhibitors of rotamase with neurotrophic effects (Lyons et al.). Based on these data, it is expected that immunosuppressants may be used to treat various peripheral neuropathies and to enhance neuronal growth in the central nervous system (CNS). Studies have shown that neurodegenerative disorders such as Alzheimer's disease, Parkinson's disease, and amyotrophic lateral sclerosis (ALS) can result from the loss or reduced availability of neurotrophic material specific to a particular population of neurons affected by the disease.

Several neurotrophic factors affecting particular populations of neurons in the central nervous system have been identified. For example, it has been suggested that Alzheimer's disease results from a reduction or loss of nerve growth factor (NGF) levels. Therefore, it has been proposed to treat patients with Alzheimer's disease with exogenous nerve growth factor or other neurotrophic proteins such as cerebral nerve factor derivative (BDNF), neuroglial factor derivative, ciliary neurotropic factor, and neurotropin-3 to enhance the fidelity of degenerate neuronal populations.

The clinical use of these proteins in a variety of neurological disease states is hampered by the difficulty of bioavailability of large proteins and delivery to the right site of the nervous system. In contrast, immunosuppressants with neurotrophic activity are relatively small and exhibit excellent bioavailability and specificity. However, long-term use has many potentially serious side effects, including nephrotoxicity such as impaired glomerular filtration and irreversible interstitial fibrosis (Kopp et al., 1991, J. Am. Soc. Nephrol. 1: 162); neurological deficits such as involuntary tremors or non-specific cerebral angina such as non-localized headaches (De Groen et al. 1987, N. Engi J. Med. 317: 861); and vascular hypertension and its complications (Kahan et al., 1989, N. Eng. J. Med. 321: 1725).

To avoid side effects associated with the use of immunosuppressive compounds, the present invention provides non-immunosuppressive compounds containing low molecular weight FKBP rotamase inhibitors for promoting neuronal growth and regeneration in a variety of neuropathophysiological situations that may facilitate neuronal repair, including peripheral nerve injury, or diseases such as diabetes, for the treatment of physical injury to the central nervous system (spinal cord and brain) associated with apoplexy and neurological disorders associated with 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 affinity for FKBP-type immunophilins. Once bound to this protein, the neurotrophic compounds are potent inhibitors of the enzymatic activity associated with immunophilin proteins, and in particular rotamase, and therefore stimulate neuronal regeneration and regrowth. A key feature of the compounds of the present invention is that they exhibit at least slightly higher immunosuppressive activity. .

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

<img file="LT4484B_D0001.tif" />

in which

R 1 is selected from the group consisting of straight or branched C 1 -C 4₉-alkyl or -alkenyl groups which may be substituted with C3-C₈-cycloalkyl, C₃ or Cs cycloalkyl, C₅-C₇«Cycloalkenyl, An, wherein said alkyl, alkenyl, cycloalkyl or cycloalkenyl groups may be substituted with C₄-alkyl, C1-C₄-alkenyl or hydroxyl, and wherein Ar_t 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-, 4-pyridyl and phenyl substituted with 1 to 3 substituents independently selected from the group consisting of hydrogen, halogen, hydroxyl, nitro, trifluoromethyl, straight or branched C 1 -C 5 alkyl or -alkenyl, C 1 -C 5₄alkoxy or Ci-C₄-alkenyloxy, phenoxy, benzyloxy and amino;

X is selected from the group consisting of oxygen, sulfur, methylene (CH 2) or H₂;

Y is selected from the group consisting of oxygen or NR₂, where R₂ is hydrogen or C1 -C₆-alkyl; and

Z is selected from the group consisting of hydrogen or straight or branched chain C₂-C 6 -alkyl or -alkenyl substituted in the alkyl chain by one or more substituents An, C as defined above₃C 6 -cycloalkyl, cycloalkyl attached to a C 1 -C 6 straight or straight-chain C 1 -C 6 alkyl or alkenyl chain, and Ar₂where 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 containing 1-3 substituents independently selected from the group consisting of hydrogen, halogen, hydroxyl, nitro, trifluoromethyl, straight or branched C1-C_e-alkyl or alkenyl, C1-C₄-alkoxy or C 1 -C 4 -alkenyloxy, phenoxy, benzyloxy and amino;

Z can also be a fragment:

O

- CH-U-X, - R₄

I r₃ »

in which

R₃ is selected from the group consisting of straight or branched C1-C₈ alkyl which may be substituted with C₃-C 1 -C 6 -cycloalkyl or An and Ari as described above, unsubstituted:

X₂ is O or NR 5 where R₅ is selected from the group consisting of hydrogen, straight or branched C 1 -C 5 alkyl or -alkenyl;

R₄ is selected from the group consisting of phenyl, benzyl, C 1 -C 5 alkyl or -alkenyl, straight or branched, and C 1 -C 5 alkyl or -alkenyl, linear or branched, substituted by phenyl; or pharmaceutically acceptable salts or hydrates thereof.

Another suitable embodiment of the present invention is a neurotrophic compound having the formula:

<img file="LT4484B_D0002.tif" />

in which

Ri is a straight or branched Ci-C chain₉-alkyl or -alkenyl groups which may be substituted with C 3 -C 6 -cycloalkyl, C 3 or C₅cycloalkyl, C₅-C₇-cycloalkenyl or Ar, wherein said alkyl, alkenyl, cycloalkyl or cycloalkenyl groups may be substituted with C1-C4 alkyl, C1-C₄-alkenyl or hydroxyl, and wherein Ari is selected from the group consisting of 1-naphthyl, 2-naphthyl, 2-indolyl, 3-indoyl, 2-furyl, 3-furyl, 2-thiazolyl, 2-thienyl, 3-thienyl, 2 -, 3- or 4-pyridyl or phenyl substituted with 1-3 substituents independently selected from the group consisting of hydrogen, halogen, hydroxyl, nitro, trifluoromethyl, straight or branched C_rC₆-alkyl or -alkenyl, Ci-C₄-alkoxy or Cr C₄-alkenyloxy, phenoxy, benzyloxy and amino;

Z is hydrogen or straight or branched C₂-C 6 -alkyl or -alkenyl substituted at one or more positions on the alkyl chain by Art, C 8 -C 6 -cycloalkyl, cycloalkyl, bonded by a straight or unbranched C 1 -C 6 -alkyl or -alkenyl chain, or Ar₂where Ar₂ is selected from the group consisting of 2-indolyl, 3-indolyl, 2furyl, 3-furyl, 2-thiazolyl, 2-thienyl, 3-thienyl, 2-, 3- or 4-pyridyl or phenyl containing 1-3 substituents independently selected from the group consisting of hydrogen, halogen, hydroxyl, nitro, trifluoromethyl, straight or branched C 1 -C 6 alkyl or -alkenyl, C 1 -C 6₄-alkoxy or C 1 -C 4 -alkenyloxy, phenoxy, benzyloxy and amino; or pharmaceutically acceptable salts or hydrates thereof.

Another suitable embodiment of the present invention is a neurotrophic compound having affinity for FKBP-type immunophilins, which inhibits the rotamase activity of the immunophilin.

Another suitable embodiment of the present invention is the use of neurotrophic compounds having affinity for FKBP-type immunophilins which inhibit the rotamase activity of immunophilin in the treatment of neurological disorders in animals.

Another suitable embodiment of the present invention is the use of neurotrophic compounds having affinity for FKBP-type immunophilins that inhibit the rotamase activity of immunophilin to promote regeneration and growth of mammalian neurons.

Another suitable embodiment of the present invention is the use of neurotrophic compounds having affinity for FKBP-type immunophilins which inhibit the rotamase activity of immunophilin in the prophylaxis of animal neurodegeneration.

Another suitable embodiment is a neurotrophic Ngroxyprolesterol having the formula:

<img file="LT4484B_D0003.tif" />

Rice in which

Rh is straight or branched CrC₅-ajkyl or -alkenyl, which may be substituted with C₃-C_e-cycloalkyl or Ari, wherein An is selected from the group consisting of 2-furyl, 2-thienyl or phenyl;

X is selected from the group consisting of oxygen and sulfur;

Y is oxygen; and *

Z'is hydrogen or straight or branched chain acyl or alkenyl substituted with Ari, Ca-C 6 -cycloalkyl, Ar in one or more positions in the alkyl chain₂where Ar₂ is selected from the group consisting of 2-, 3-, or 4-pyridyl and phenyl substituted with 1-3 substituents independently selected from the group consisting of hydrogen and C 1 -C 6;₄-alkoxy groups.

Particularly suitable neurotrophic N -glyoxylprolyl esters of the above formula are esters selected from the group consisting of:

3- (2,5-dimethoxyphenyl) -1-propyl (2S) -1- (3,3-dimethyl-1,2-dioxopentyl) -2-pyrrolidinecarboxylate,

3- (2,5-dimethoxyphenyl) -1-prop-2- (E) -enyl- (2S) -1- (3,3-dimethyl-1,2-dioxopentyl) -2-pyrrolidinecarboxylate,

2- (3,4,5-ΐπηηθίοΚ8ϊΐ © ηί |) ~ 1- © ΐϋ- (23) -1- (3,3-ι3ΐηηβίϋ-1,2-dioxopentyl) -2-pyrrolidinecarboxylate,

3- (3-pyridyl) -1-propyl (2S) -1- (3,3-dimethyl-1,2-dioxopentyl) -2-pyrrolidinecarboxylate,

3- (2-pyridyl) -1-propyl (2S) -1- (3,3-dimethyl-1,2-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-tert-butyl-1,2-dioxoethyl) -2-pyrrolidinecarboxylate,

3-phenyl-1-propyl- (2 S) -1- (2-cyclohexyl ethyl-1,2-dioxoethyl) -2-pyrrole dinecarboxylate,

3- (3-pyridyl) -1-propyl (2S) -1- (2-cyclohexylethyl-1,2-dioxoethyl) -2-pyrrole dinecarboxylate,

3- (3-pyridyl) -1-propyl (2S) -1- (2-tert-butyl-1,2-dioxoethyl) -2-pyrrolidinecarboxylate,

3.3-diphenyl-1-propyl (2S) -1- (3,3-dimethyl-1,2-dioxopentyl) -2-pyridinecarboxylate,

3- (3-pyridyl) -1-propyl- (2S) -1- (2-cyclohexyl-1,2-dioxoethyl) -2-pyrrolidine carboxylate,

3- (3-Pyridyl) -1-propyl (2S) -N - ([2-thienylglyoxyl) pyrrolidinecarboxylate, 3_l3-diphenyl-1-propyl (2S) -1- (3,3-dimethyl-1,2-dioxobutyl) -2-pyrrolidinecarboxylate,

3.3-Diphenyl-1-propyl- (2S) -1-cyclohexylglyoxyl-2-pyrrolidinecarboxylate; and

3.3-Diphenyl-1-propyl- (2S) -1- (2-thienyl) glyoxyl-2-pyrrolidinecarboxylate.

BRIEF DESCRIPTION OF THE FIGURES

Figure 1 is a photomicrograph of the chick dorsal root ganglia exposed to various concentrations of the compound described in Example 17. Figure 1 shows that the compound of Example 17 of the present invention greatly promotes axonal growth in cultures of sensory neurons. Cultures of explants (tissues taken from the body) isolated from the dorsal root ganglia of chick embryos for 9 to 10 days were exposed to various concentrations of the compound described in Example 17 as shown in the figure. After 48 hours, the number of axons with more than one DRG explant length was determined. The number of axons expressed in untreated DRGs was subtracted from the number of axons of the sample treated with Example 17 compound and the specific axon growth dependence of Example 17 compound was obtained. Micrographs of DRGs exposed to the compound of Example 17 as well as quantitative dose-dependency of axon growth induced by the compound of Example 17 are shown,

Figure 2 is a graph showing axonal growth in the dorsal root ganglia of chickens exposed to various concentrations of the compound described in Example 17 as shown in Figure. Figure 2 shows that the compound of Example 17 of the present invention strongly promotes axonal growth in cultures of sensory neurons. Expert cultures isolated from the posterior spinal cord ganglia of chick embryos for 9 to 10 days were exposed to various concentrations of the compound described in Example 17 as shown in the figure. After 48 hours, the amount of axons with more than one DRG explant length was determined. The number of axons expressed in untreated DRGs was subtracted from the number of axons in the sample treated with Example 17 compound and the specific axon outgrowth from Example 17 compound was obtained. A quantitative dose-dependence of axonal growth induced by the compound of Example 17 is shown.

Figure 3 is a photomicrograph of sections of the rat sciatic nerve. Figure 3 shows that the compound of Example 1 of the present invention promotes neuronal regeneration following sciatic nerve injury. Male Sprague-Dawley rats (150 g) had sciatic nerve at hip level. Thereafter, Example 1 (30 mg / kg, sc), inactive compound (30 mg / kg, sc) or intralipidic solvent was administered once daily for 21 days. The animals were sacrificed, the sciatic nerve removed, incised at 2 mm from the nerve injury site and stained with Holmes silver dye (for axon counts) and Luxol rapid blue (for remyelination). Micrographs show sections of the sciatic nerve in a false-exposed rat, solvent-exposed lesioned animals, Example 1 compound, and inactive compound-exposed animals at 630x magnification (4 animals per group).

Fig.4 is [³The graph of H 1 -CFT binding (micrograms) to the striped membrane protein of Fig. 4 shows that the neuroimmunophilinic ligands of the present invention promote the recovery of dopamine neurons after exposure to MPTP in mice. CD1 mice (25 g) were exposed to 30 mg / kg MPTP (ip) daily for five days. Animals were also daily treated with intralipidic solvent, Example 1 compound (100 mg / kg, sc) or Example 17 compound (40, 20, 10 mg / kg, sc as indicated) in combination with MPTP and continued for another 5 days. After 18 days, the mice were sacrificed, stripped, and streaked in the group of 5 animals and transformed into a washed membrane preparation. Has been quantified [³H] -CFT binding to various classes of these striped-body membrane preparations to determine amounts of dopamine transporter to viable nerve endings. Binding in the presence of 10 μΜ of unlabeled CFT allowed for non-specific binding to be subtracted from total binding by assessing specific [³H] -CFT binding. Binding was normalized to protein content of striped body membranes in each experimental group. Coronary and sagittal sections of MPTP and drug-treated animals were stained with anti-tyrosine hydroxylase (TH) Ig to determine TH levels in the striated body, in the axons of the medial forebrain, and in the black substance, which show functional dopaminergic neurons.

Fig.5 is In³H] -CFT binding diagram for 200 μg membrane protein column ©. Figure 5 shows that the neuroimmunophilic ligands of the present invention promote the recovery of dopamine neurons following mouse MPTP treatment according to the procedure described in Figure 4,

Figure 6 is a photomicrograph of coronary and sagittal sections of the brain. at 630x magnification. Fig. 6 shows brain sections of MPTP and drug-treated animals stained with anti-tyrosine hydroxylase (TH) Ig to determine TH levels in the striped body, which show functional dopaminergic neurons.

Figure 7 is a photomicrograph of coronary and sagittal sections of the brain at 50X magnification. Figure 7 shows brain sections of MPTP and drug-treated animals stained with anti-tyrosine hydroxytase (TH) Ig to determine TH levels in the black substance, which show functional dopaminergic neurons.

Figure 8 is a photomicrograph of coronary and sagittal sections of the brain at 400x magnification. Figure 7 shows brain sections of MPTP and drug-treated animals stained with anti-tyrosine hydroxylase (TH) Ig for the determination of TH levels in medial forebrain fiber axons, which show functional dopaminergic neurons.

DETAILED DESCRIPTION OF THE INVENTION

The novel neurotrophic compounds of the present invention are relatively small molecules that bind to FKBT-type immunophilins such as rapamycin, FK506, and cyclosporine, compared to other known compounds.

The neurotrophic compounds of the present invention have affinity for FK506 binding proteins such as FKBP-12. Unexpectedly, when the neurotrophic compounds of the present invention are bound to FKBP, they inhibit the prolyl-peptidyl-cis-trans -isomerase activity or rotamase activity of the binding protein and stimulate axonal growth, but do not possess immunosuppressive activity.

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

W^Y~^Z ° γλχ ° ^R'in which

Ri is a straight or branched chain C »-C₉-alkyl or -alkenyl group which may be substituted with C₃-C 8 -cycloalkyl, C₃ or C₅ cycloalkyl, C₅C 2 -cycloalkenyl or Ari wherein said alkyl, alkenyl, cycloalkyl or cycloalkenyl groups may be substituted with C₄-alkyl, CrC₄-alkenyl or hydroxyl and wherein Ari is selected from the group consisting of 1-naphthyl, 2-naphthyl, 2-indolyl, 3-indolyl, 2-furyl, 3-furio, 2-thiazolite, 2-thienyl, 3-thienyl, 2-, 3- or 4-pyridyl or phenyl substituted with 1-3 substituents independently selected from the group consisting of hydrogen, halogen, hydroxyl, nitro, trifluoromethyl, straight or branched C 1 -C 6 alkyl or -alkenyl, C 1 -C₄-alkoxy groups or Ci-C₄-alkenyloxy, phenoxy, benzyloxy and amino;

X is oxygen, sulfur, methylene (CH₂) or H₂;

Y is oxygen or NR₂, where R₂ is hydrogen or C 1 -C 6 alkyl; and

Z is hydrogen or straight or branched C2-C₃-alkyl or -alkenyl substituted at one or more positions on the alkyl chain with An, C as defined above₃-C₈-cycloalkyl, cycloalkyl, bonded in a straight or unbranched C1-C₆-alkyl or alkenyl chain, or Ar₂where Ar₂ is selected from the group consisting of 2-indolyl, 3-indolyl, 2furyl, 3-furyl, 2-thiazolyl, 2-thienyl, 3-thienyl, 2-, 3- or 4-pyridyl and phenyl containing 1-3 substituents independently selected from the group consisting of hydrogen, halogen, hydroxy, nitro, trifluoromethyl, straight or branched C 1 -C 6 alkyl or -alkenyl, C 1 -C₄-exoxy groups or Cr C₄-afenyloxy, phenoxy, benzyloxy and amino:

Z can also be a fragment:

o

-CH —- X₂- R₄ r₃

I create

R 3 is selected from the group consisting of straight or branched C 1 -C 5 -alkyl which may be substituted with C₃-C 8 -cycloalkyl or Ari and Ari unsubstituted as defined above;

Χ2 is O or NR₅₎ wherein R 5 is selected from the group consisting of hydrogen, straight-chain or branched C 1 -C 6 alkyl or -alkenyl;

R₄ is selected from the group consisting of phenyl, benzyl, C 1 -C 5 alkyl or -alkenyl straight or branched, and C 1 -C 5 straight or branched₅-alkyl or -alkenyl substituted with phenyl; or pharmaceutically acceptable salts thereof, or hydrates thereof.

More preferred compounds are those of the formula:

<img file="LT4484B_D0004.tif" />

R1 wherein

R 1 is a straight or branched C 1 -C 8 -alkyl or -alkenyl group which may be substituted with C₃-C 8 -cycloalkyl, C₃ or C₅ cycloalkyl, C₅C₇-cycloalkenyl or An, wherein said alkyl, alkenyl, cycloalkyl or cycloalkenyl groups may be substituted with CrC₄-aIkil, CrC₄-alkenyl or hydroxyl, and wherein Ari 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 or phenyl substituted with 1-3, which are independently selected from the group consisting of hydrogen, halogen, hydroxy, nitro, trifluoromethyl, straight or branched C1-C₆alkyl or -alkenyl, CrC₄-alkoxy or Ci-C₄-alkenyloxy, phenoxy, benzyloxy and amino;

Z is hydrogen or straight or branched C₂-C₆-alkyl or -alkenyl, in which the alkyl chain has one or more substituents aukščiauη, C as defined above at one or more positions₃-C₈-cycloalkyl, cycloalkyl, bonded straight or unbranched C1-C₆-alkyl or -alkenyl chain, or Ar₂where Ar₂ is selected from the group consisting of 2-indolyl, 3-indolyl, 2-furyl, 3furyl, 2-thiazolyl, 2-thienyl, 3-thienyl, 2-, 3- or 4-pyridyl or phenyl containing 1-3 substituents independently selected from the group consisting of hydrogen, halogen, hydroxyl, nitro, trifluoromethyl, straight or branched C1-C₆-alkyl or -alkenyl, C1-C₄-alkoxy or CiC₄-alkenyloxy, phenoxy, benzyloxy and amino; or pharmaceutically acceptable salts thereof, or hydrates thereof.

Suitable neurotropic N-glyoxylprolyl esters have the formula:

in which

Ri is a straight or branched Ci-C chain₅-alkyl or -alkenyl group which may be substituted with C₃-C₆-cycloalkyl, or An where Ar! is selected from the group consisting of 2-furyl, 2-thienyl or phenyl;

X is selected from the group consisting of oxygen and sulfur;

Y is oxygen; and

Z is hydrogen or straight or branched alkyl or alkenyl substituted in one or more positions on the alkyl chain with t

is as described above Ar, C₃-C₆-cycloalkyl, Ar_2l where Ar₂ is selected from the group consisting of 2-, 3- or 4-pyridyl and phenyl substituted with 1-3 independently selected from the group consisting of hydrogen and C 1 -C 4 alkoxy.

The compounds of the present invention exist in the stereoisomeric forms, which are either enantiomers or diastereoisomers. The stereochemistry at position 1 (Formula 1) is R or S; The present invention includes enantiomers, racemic forms and mixtures of diastereoisomers. Enantiomers and diastereoisomers may be separated from one another by methods known to those skilled in the art.

Immunophilins such as FKSP are known to best recognize peptide substrates containing Xaa-Pro-Yaa fragments in which Xaa and Yaa are lipophilic amino acid residues (Schreiber et al., 1990, J. Org. Chem., 55, 4984- 4986; Harrison and Stein, 1990, Biochemistry 29, 3813-3816). Thus, modified prolyl-peptide-imitator compounds having lipophilic substituents should bind with high affinity to the hydrophobic portion of the FKBP active site and inhibit its rotamase activity.

The preferred compounds of the present invention are those wherein the Ri group is not stereochemically very large in volume relative to the known size and shape of the hydrophobic moiety of the FKBP active site.

Preferred compounds of the present invention are:

(23) -1- (1,2-dioxo-3,3-dimethylpentyl) -2-pyrrolidinecarboxylic acid,

3-phenyl-1-propyl- (2S) -1- (3,3-dimethyl-1,2-dioxopentyl) -2-pyrrolidinecarboxylate,

3-phenyl-1-prop-2- (E) -enyl- (2S) -1- (3,3-dimethyl-1,2-dioxopentyl) -2-pyrrolidinecarboxylate,

3- (3,4,5-trimethoxyphenyl) -1-propyl- (2S) -1- (3,3-dimethyl-1,2,2-dioxopentyl) -2-pyrrolidinecarboxylate,

3- (3,4,5-trimethoxyphenyl) -1-prop-2- (E) -enif- (2S) -1- (3,3-dimethyl-1,2-dioxopentyl) -2-pyrrolidinecarboxylate,

3-(4_l5-methylenedioxyphenyl) -1-propyl - (2S) -1- (3,3-dimethyl-1,2-dioxopentyl) 2-pyrrolidinecarboxylate,

3- (4,5-methylenedioxyphenyl) -1-prop-2- (E) -enyl- (2S) -1- (3,3-dimethyl-1,2-dioxopentyl) -2-pyrrolidinecarboxylate,

3-cyclohexyl-1-propyl- (2S) -1- (3,3-dimethyl-1,2-dioxopentyl) -2-pyrrolidinecarboxylate,

3-οίΚΙοΚθΚ8ίΜ ^ Γθρ ^ - (Ε) -βπίΙ- (28) -1- (3,3-ίίωθΙίΜ₍2- (ΐΐΚΚΚοοοοοίΙΙ) -2-pyrrolidinecarboxylate, (1R) -1,3-diphanyl-1-propyl- (2S) -1- (3,3-dimethyl-1,2-dioxopentyl) -2-pyrrolidinecarboxylate,

3-phenyl-1-propyl (2S) -1- (1,2-dioxo-2- [2-furanyl]) ethyl 1-2-pyrrolidinecarboxylate,

3-phenyl-1-propyl- (2S) -1- (1,2-dioxo-2- [2-thienyl]) ethyl 1,2-pyrrolidinecarboxylate,

3'-phenyl-1-propyl - (2S) -1- (1,2-dioxo-2- [2-thiazolyl]) ethyl -2-pyrrolidinecarboxylate,

3-phenyl-1-propyl- (2S) -1- (1,2-dioxo-2-fanyl) atyl-2-pyrrolidinecarboxylate,

3- (2,5-dimethoxyphenyl) -1-propyl- (2S) -1- (3,3-dimethyl-1,2-dioxopentyl) -2-pyrrolidinecarboxylate,

3- (2,5-Dimethoxyphenyl) -1-prop-2- (E) -enyl- (2S) -1- (3,3-dimethyl-1,2-dioxopentyl) -2-pyrrolidinecarboxylate,

2- (3,4,5-trimethoxyphenyl) -1-ethyl- (2S) -1- (3,3-dimethyl-1,2-dioxopentyl) -2-pyrrolidinecarboxylate,

3- (3-pyridyl) -1-propyl · (2S) -1- (3,3-dimethyl-1,2-dioxopentyl) -2-pyrrolidinecarboxylate,

3- (2-pyridyl) -1-propyl- (2S) -1- (3,3-dimethyl-1,2-dioxopentyl) -2-pyrrole dincarboxylate,

3- (4-pyridyl) -1-propyl- (2S) -1- (3-_l3-dimethyl-1,2-dioxopentyl) -2-pyrrolidinecarboxylate,

3-phenyl-1-propyl- (2S) -1- (2-cyclohexyl-2-dioxoethyl) -2-pyrrolidinecarboxylate,

3-Phenyl-1-propyl (2S) -1- (2-tert-butyl-1,2-dioxoethyl) -2-pyrrole dicarboxylate,

3-phenyl-1-propyl - (2S) -1- (2-cyclohexylethyl-1,2-dioxoethyl) -2-pyrrolidinecarboxylate,

3- (3-Pyridyl) '1-prapyl- (2S)' 1- (2-cyclohexylethyl-1,2-dioxoethyl) -2-pyrrolidinecarboxylate.

3- (3-pyridyl) -1-propyl- (2S) -1- (2-tert-butyl-1,2-dioxoethyl) -2-pyrrolidinecarboxylate,

3.3-diphenyl-1-propyl- (2S) -1- (3,3-dimethyl-1,2-dioxopentyl) -2-pyrrolidinecarboxylate,

3- (3-pyridyl) -1-propyl (2S) -1- (2-cyclohexyl-1,2-dioxoethyl) -2-pyrrolidinecarboxylate,

3- (3-pyridyl) -1-propyl- (2S) -N - ([2-thienyl] glyoxyl) pyrrolidinecarboxylate,

3.3-Diphenyl-1-propyl- (2S) -1- (3,3-dimethyl-1,2-dioxobutyl) -2-pyrrolidinecarboxylate,

3.3-Diphenyl-1-propyl- (2S) -1-cyclohexylglyoxyl-2-pyrrolidinecarboxylate and 3,3'-diphenyl-1-propyl- (2S) -1- (2-thienyl) glyoxyl-2-pyrrolidinecarboxylate, Highly Suitable Neurotrophic The N-glyoxytopropyl esters are selected from the group consisting of:

3- (2,5-dimethoxyphenyl) -1-propyl (2S) -1- (3,3-dimethyl-1,2-dioxopentyl) -2-pyrrolidinecarboxylate,

3-(2_t5-dimethoxyphenyl) -1-prop-2- (E) -enyl- (2S) -1- (3,3-dimethyl-1,2-dioxopentyl) -2-pyrrolidinecarboxylate,

3-(3,4_I5-trimethoxyphenyl) -1-ethyl- (2S) -1- (3,3-dimethyl-1,2-dioxopentyl) -2-pyrrolidinecarboxylate,

3- (3-pyridyl) -1-propyl (2S) -1- (3,3-dimethyl-1,2-dioxopentyl) -2-pyrrolidinecarboxylate,

3- (2'-pyridyl) -1-propyl- (2S) -1- (3,3-dimethyl-1,2-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-tert-butyl-1,2-dioxoethyl) -2-pyrrolidinecarboxylate,

3-phenyl-1-propyl (2S) -1- (2-cyclohexylethyl-1,2-dioxoethyl) -2-pyrrolidinecarboxylate,

3 '(3-pyridyl) -1-propyl (2S) -1- (2-cyclohexyllethyl-1,2-dioxoethyl) -2-pyrrolidinecarboxylate,

3- (3-pyridyl) -1-propyl (2S) -1- (2-tert-butyl-1,2-dioxoethyl) -2-pyrrolidinecarboxylate,

3,3-diphenyl-1-propyl (2S) -1- (3,3-dimethyl-1,2-dioxopentyl) -2-pyrrole dinecarboxylate,

3- (3-pyridyl) -1-propH- (2S) -1- (2-cyclohexyl-1,2-dioxoethyl) -2-pyrrolidinecarboxylate,

3- (3-pyridyl) -1-propyl (2S) -N - ([2-thienyl] glyoxyl) pyrrolidinecarboxylate,

3,3-diphenyl-1-propyl (2S) -1- (3,3-dimethyl-1,2-dioxobutyl) -2-pyrrolidinecarboxylate,

3,3'-diphenyl'1-propyl '(2S) -1-cyclohexylglyoxyl-2-pyrrolidinecarboxylate and 3_l3-Diphenyl-1-propyl- (2S) -1- (2-thienyl) glyoxyl-2-pyrrolidinecarboxylate.

The compounds of the present invention may be used in the form of salts with inorganic or organic acids and bases. The salts with acids include the following salts: acetate, adipate, alginate, aspartate, benzoate, benzenesulfonate, bisulfate, butyrate, citrate, camparate, camphor sulfonate, cyclopentanpropionate, digluconate, dodecyl sulfate, ethanesulfonate, fumarate, glucoheptanoate, glycerophosphate, hemisulfate, hemisulfate, hemisulfate, hemisulfate, , 2-hydroxy ethanesulfonate, lactate, mayea, methanesulfonate, 2-naphthalenesulfonate, nicotinate, oxalate, pamoate, pectinate, propionate, succinate, tartrate, thiocyanate, tosylate and undecanoate. Salts with bases include ammonium salts, alkali metal salts such as sodium and potassium salts, alkaline earth metal salts such as caffeine and magnesium salts, organic salts such as dicyclohexylamine salts, N-methyl-D-glucamine, and salts with amino acids such as arginine, lysine and so on. In addition, basic nitrogen-containing groups may be quaternized with agents such as lower alkyl halides such as methyl, ethyl, propyl and butyl chlorides, bromides and iodides; dialkyl sulfates such as dimethyl, diethyl, dibutyl and diamyl sulfates; long-chain halides such as decyl, lauryl, myristyl and stearyl chlorides, bromides and iodides; aralkyl halides such as benzyl and phenethyl bromides and others. This results in water or oil soluble or dispersible products.

The neurotrophic compounds of the present invention may be periodically administered to a patient undergoing treatment for a neurological disorder, or for other reasons where it is desirable to stimulate neuronal regeneration and growth, such as various peripheral neuropathic and neurological disorders associated with neurodegeneration. The compounds of the present invention may also be administered to other non-human mammals for the treatment of a variety of neurological disorders in a mammal.

The novel compounds of the present invention are potent inhibitors of rotamase activity and exhibit an excellent degree of neurotrophic activity. This activity is useful for the stimulation of damaged neurons, the promotion of neuronal regeneration, the prevention of neurodegeneration, and the treatment of several neurological disorders that are associated with neuronal degeneration and peripheral neuropathies. Neurological disorders that may be treated include, but are not limited to: trigeminal nerve pain, glossopharyngeal neuralgia, Belle's palsy, myasthenia gravis, muscular dystrophy, lateral amyotrophic sclerosis, progressive bulbar hereditary muscle atrophy, intervertebral disc herniation, rupture or fall out syndrome, cervical spondylitis, cervical spondylosis, caused by lead, diaminodiphenylsulfone (dapsone), mites, porphyria or Gulen-Barre syndrome, Alzheimer's disease and Parkinson's disease.

For these purposes, the compounds of the present invention may be administered orally and parenterally, by inhalation injection, topically, rectally, intranasally, intramuscularly, or vaginally, or via implanted drug sources in dosage compositions containing conventional non-toxic pharmaceutically acceptable _t suitable carriers, stimulants, and solvents. As used herein, the term parenteral includes subcutaneous, intravenous, intramuscular, intraperitoneal, intraventricular, intraventricular, intrasternal, and intracranial injection or infusion techniques.

In order to be therapeutically effective as a target for the central nervous system, the immunophilin-drug complex must readily cross the blood-brain barrier if the drug is administered peripherally. Compounds of the present invention that cannot penetrate the blood-brain barrier can be effectively administered intraventricularly.

The pharmaceutical compositions may be in the form of a sterile injectable preparation, for example, as a sterile aqueous or greasy suspension for injection. This suspension may be formulated according to techniques known 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 parenteral diluent or solvent, such as a solution in 1,3-butanediol. Suitable diluents and solvents that may be used include water, Ringer's solution and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. Any mild, non-volatile oil can be used for this purpose, including synthetic mono- and diglycerides. Fatty acids such as oleic acid and its glycerides find use in olive oil or castor oil preparations for injection, in particular in their polyoxyethylated versions. These oily solutions or suspensions may also contain long chain alcohol as a diluent or dispersant.

These compounds may be administered orally, for example in the form of tablets or capsules, or as suspensions or solutions in water. In the case of tablets for oral use, the most commonly used carriers are lactose and maize starch. Typically, lubricating agents such as magnesium stearate are also added in the form of oral diluents in the form of capsules containing lactose and dried corn starch. If the oral suspension requires aqueous suspensions, the active ingredient is mixed with emulsifiers and suspending agents. Certain sweeteners and / or flavoring and / or coloring agents may be added.

The compounds of the present invention may also be used in the form of suppositories for rectal administration. These suppositories can be made by mixing the drug with a suitable non-irritating excipient which is solid at room temperature but liquid at rectal temperature and therefore melts in the rectum as the drug is released. Such materials include cocoa butter, beeswax and polyethylene glycols,

The substances of the present invention may be administered topically, in particular for the treatment of areas or organs readily accessible for topical administration, including neurological disorders of the eye, the skin or the lower gastrointestinal tract. Each of these areas can easily be formulated into topical formulations.

For administration to the eye, the compounds may be formulated as suspensions of micron particles in isotonic, pH-adjusted saline, or more preferably, as solutions in isotonic, pH-adjusted saline, with or without preservatives such as benzylalkonium chloride. Alternatively, for administration to the eye, the compounds may be formulated in an ointment such as petroleum jelly.

For topical application to the skin, the compounds may be formulated in a suitable ointment in which the compound is suspended or dissolved, for example, in a mixture of one or more of the following: mineral oil, liquid petroleum jelly, light petroleum jelly, propylene glycol, polyoxyethylene-polyoxypropylene emulsifier and water. . Alternatively, the compounds may be formulated in a suitable lotion or cream in which the active compound is suspended or dissolved, for example in a mixture of one or more of the following: mineral oil, sorbitan monostearate, polysorbate 60, cetyl ester wax, cetearyl alcohol, 2-octyldodecanol. , benzyl alcohol and water.

Topical administration to the lower gastrointestinal tract may be effected by the use of renal suppository compositions (see above) or by enema formulations.

Suitable doses for treating the above conditions are from about 0.1 mg to about 10,000 mg of active ingredient, preferably from about 0.1 to about 1000 mg per turn. The amount of active ingredient that can be mixed with the carriers in individual dosage forms will vary depending on the nature being treated and the particular mode of administration.

However, it should be understood that the individual patient's dosage will depend on a variety of factors including the activity of the particular compound being administered, the patient's age, body weight, general health, sex, food, time of administration, excretion rate, drug combination, acutely specific disease and drug use. way.

These compounds can be used with other neurotrophic agents such as neurotrophic growth factor (NGF), growth factor as a neuroglial derivative, growth factor as a brain derivative, ciliary neurotrophic factor, and neurotropin-3. The dosage of other neurotrophic drugs will depend on the above factors and on the neurotrophic efficacy of the combination.

Methodology for the determination of K

Inhibition of peptidyl-prolylisomerase (rotamase) activity by the compounds of the invention can be evaluated by methods known in the art (Harding, MW et al., Nature 341: 758-760 (1989): Hoft et al., J. Am. Chem. Soc. 115: 9923). -9938). These values ​​are derived as the implicit K and are given in Table I. The cis-trans isomerization of the alanine-proline bond in a model substrate, N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide, was controlled spectrophotometrically in a chymotrypsin assay, in which para-nitroaniline is separated from the substrate transform. The inhibition of this reaction by the addition of various concentrations of the inhibitor is determined and the data analyzed as a function of the first-order rate constant change from the inhibitor concentration from which the apparent K i values ​​are obtained. j Plastic cuvette {add 950 μΙ. ice-cooled assay buffer (25 mM HEPES, pH 7.8,100 mM NaCl), 10 μΙ FKBP (2.5 mM solution in 10 mM Tris-Cl, pH 7.5,100 mM NaCl, 1 mM dithiothrietol), 25 μΙ chymotrypsin (50 mg / mL in 1 mM HCl solution) and 10 μ! various concentrations of the test compound in dimethylsulfoxide. The reaction was initiated by addition of 5 μΙ of substrate (succinylAla-Phe-Pro-Phe-para-nitroanilide, 5 mg / ml in 2.35 mM UGI in trifluoroethanol).

The spectrophotometer measures the time dependence of the optical density at 390 nm for 90 seconds and determines the rate constants from the time dependence of the optical density.

The data for these experiments are shown in Table I.

Table I

No.

R

1,1-dimethylpropyl

<img file="LT4484B_D0005.tif" />

K

11 12 .

21 22

2-furanyl

2-thienyl

2-Thiazolyl phenyl

1,1-dimethylpropyl cyclohexyl tert-butyl cyclohexylethyl cyclohexylethyl tert-butyl

1,1-dimethylpropyl cyclohexyl

3-phenylprop-2- (E) -enyl

3- (3,4,5-trimethoxyphenyl) propyl

3- {3A5-trimethoxyphenyl) prop-2 (E) -enyl

3- (4,5-methylenedioxy) phenylpropyl

3- (4,5-methylenedioxy) phenyl-prop-2 (E) -enyl

3-Cyclohexylpropyl 3-Cyclohexylprop-2- (E) -enyl (1R) -1,3-diphenyl-1-propyl 3-phenylpropyl

3- (2,5-Dimethoxy) phenylpropyl

3- {2,5-dimethoxy) phenylprop-2- (E) -enyl

2- (3,4,5-trimethoxyphenyl) ethyl

3- (3-pyridylpropyl) 3- (2-pyridyl) propyl 3- (4-pyridyl) propyl 3-phenylpropyl

3- (3-pyridyl) propyl

3- (3-pyridyl) propyl

3,3-diphenylpropyl

3- (3-pyridyl) propyl

125

200

160

100

450

1025

1400

170

200

600

4000

1970

250

120

195

<td> 27</td><td>2-thienyl</td><td>3 '(3-pyridyl) propyl</td><td></td>

<td> 28</td><td>tert-butyl</td><td>3,3-diphenylpropyl</td><td> 5</td>

<td> 29</td><td>cyclohexyl</td><td>(i</td><td> 20</td>

<td> 30</td><td>2-thienyl</td><td>U</td><td></td>

In mammalian cells, FKBP-12 complexes with the inositol triphosphate receptor (IP3R) and the ryanodine receptor (RyR). The compounds of the present invention are believed to cleave FKBP-12 from these complexes, causing the calcium channel to become "leaky" (Cameron et al., 1995). Calcium efflux causes axonal extension, and therefore the IP3R receptor and the ryanodine receptor may be involved in the neurotrophic effects of the drug. Because the drug binds to the same site as FKBP-12 as the IP3R receptor, it can be assumed that the drug removes FKBP-12 in the channels.

In the chicken, the dorsal root ganglia of the chicken

Cultures and axonal growths

The posterior dorsal root ganglia (DRG) were separated from the embryos of ten-day-old chicks. Full ganglion explants were cultured on thin-layer matrigel-coated 12-well plates with Liebovitz L15 plus glucose-rich medium containing 2 mM glutamine and 10% calf fetal serum plus 10 μΜ of cytosine β-D arabinofuranoside (Ar) 37 ° C and an environment containing 5% CO2. After 24 hours, DRGs were exposed to various concentrations of nerve growth factor, immunophilin ligands, or NFG plus drug combinations. 48 hours after drug treatment, the ganglia were visualized by phase contrast or Hofmann modulation contrast using a Zeiss Axiovert inverse microscope. Exploratory photomicrographs were performed and axonal regimens were quantified. Axons longer than the DGR diameter were counted as positive by determining the total number of axons. evaluated for each experimental condition. 3-4 wells were cultured in each well and each treatment was repeated twice.

The data for these experiments are shown in Table II. Typical photomicrographs of 17 examples are shown in FIG. the dose-response curve for this example is given in FIG.

Table II

Axon graft in chicken DRG Fig, Nr, EDsg, axon graft in nM

<td> 1</td><td> 53</td>

<td> 2</td><td> 105</td>

<td> 3</td><td> 149</td>

<td> 4</td><td> 190</td>

<td> 5</td><td> 10</td>

<td> 6</td><td> 75</td>

<td> 10</td><td> 0,46</td>

<td> 11</td><td> 0,015</td>

<td> 14</td><td> 2</td>

<td> 15</td><td> 0,8</td>

<td> 16</td><td> 0,015</td>

<td> 17</td><td> 0,05</td>

<td> 18</td><td> 30</td>

<td> 19</td><td> 6</td>

<td> 20</td><td> 0,13</td>

<td> 21</td><td> 0,025</td>

<td> 22</td><td> 0,66</td>

<td> 23</td><td> 1100</td>

<td> 24</td><td> 0,014</td>

<td> 25</td><td> 0,50</td>

<td> 26</td><td> 2</td>

<td> 27</td><td> 500</td>

<td> 28</td><td> 0,50</td>

<td> 29</td><td> 10</td>

<td> 30</td><td> 100</td>

Axotomy of the sciatic nerve

Six-week-old Sprague-Davvley rats were:. t anesthetized, and nude and tweezed their sciatic nerve at hip level. Test compounds or vehicle were injected subcutaneously immediately before and daily after the lesion. Sciatic nerve sections were stained with Holmes silver for axon quantification and Luxol fast blue myelin quantification. Eighteen days after lesion, a significant reduction (50% decrease in axon count) and a decrease in myelination rate were observed in solvent-treated animals ( 90% reduction compared to intact control rats), sample compound administration (30 mg / kg, (sc) gave a significant increase in both axon counts (5% reduction compared to intact control rats) and myelination rate (50% reduction compared to controls) immediately before and once daily for 18 days after lesion, animals, regeneration. This significant efficacy of the compound of Example 1 is consistent with its clearly expressed activity in inhibiting rotamase activity and stimulating axon sprouts in chicken DRGs. These results are shown in Fig. 3. "Deception" refers to control animals that received the solvent but were not injured; "Solvent" refers to animals that have been damaged and received only solvent (ie no drug is present). Exposure to the compound of Example 1 showed remarkable similarity to the trick animals, indicating that these compounds exhibit potent neuroregenerative activity in vivo. Inactive is the compound which is inactive as an inhibitor of FKBP-12. Animals exposed to such a compound were similar to solvent-exposed lesioned animals; this confirms that the neuroregeneration results observed for the compound of Example 1 are directly induced by the inhibition of FKBP-12 by this compound. The quantitative evaluation of these data is shown in Table lll.

Table III

<td>Impact</td><td>Number of axons (% of control)</td><td>Loved it</td>

<td>The trick</td><td> 100</td><td> 100</td>

<td>Violation:</td><td></td><td></td>

<td>+ solvent (sc)</td><td> 50</td><td> 10</td>

<td>+ Compound of Example 1 (30 mg / kg,</td><td> 100</td><td> 50</td>

<td>sc)</td><td></td><td> *</td>

<td>+ inactive compound (30 mg / kg, sc)</td><td> 25</td><td> 25</td>

MPTP model of Parkinson's disease in a mouse mouse MPTP lesion in mouse dopaminergic neurons was used as an animal model of Parkinson's disease. Male CD1 white mice at 4 weeks of age were ip injected with 30 mg / kg MPTP for 5 days. The compound of Example 17 (10-40 mg / kg) was administered sc for 5 days with MPTP and for 5 days after MPTP was stopped. 18 days after exposure to MPTP, the animals were sacrificed and separated and their striped bodies homogenized. To determine dopamine transporter (DAT) levels after injury and after drug exposure, [³Coupling of H] CFT (Radioligand for Dopamine Transporter) to Striped Body Membranes. Immunostaining of sagittal and coronary brain sections using anti-tyrosine hydroxylase Ig was performed to determine the amount of survival and recovery of dopaminergic neurons. In animals exposed to MPTP and solvent, significant loss of functional dopaminergic terminal neurons was observed compared to intact animals. Nearly quantitative recovery of TH-stained dopaminergic neurons was observed in injured animals receiving Compound 17.

Figures 4 and 5 show a quantification of DAT levels, and Figures 6-8 are photomicrographs showing the regenerative effect of Example 17 in this model. Fig. 4 shows a marked recovery of functional dopaminergic terminal neurons as determined from [³H] -CFT binding assay compared to animals receiving MPTP but not Guilford compounds. Fig. 5 shows this data in the form of a bar chart. The animals that received 40 mg / kg of the compound of Example 17 together with MPTP were shown to clearly show greater than 90% [³H] -CFT binding recovery. As shown in Figures 6-8, immunostaining with tyrosine hydroxylase (a marker for viable dopaminergic neurons) in the striped body, black substance, and medial forebrain shows clear and marked recovery of functional neurons. agent but did not receive the drug (MPTP / solvent),

The following examples are illustrative of preferred embodiments of the present invention and should not be construed as a limitation of the invention. All preferred embodiments of the present invention are not to be construed as limitations. All polymers have average molecular weights. All percentages are based on percentages based on the final weight of the system or composition being prepared, unless otherwise stated, and the total is 100% by weight.

EXAMPLES The compounds of the present invention can be prepared according to various synthetic sequences using known chemical transformations. A general procedure for preparing these compounds is depicted in Scheme I. The N-glyoxylproline derivatives can be prepared by reacting the methyl ester of L-proline with methyloxalyl chloride as shown in Scheme I. The resulting oxamates can be exposed to various carbon nucleophiles to form intermediates. These intermediates are then treated with various alcohols, amides, or blocked amino acid residues to give the propyl esters and amides of the present invention.

scheme

U

H

<img file="LT4484B_D0006.tif" />

OCH,

RU or RMgX

<img file="LT4484B_D0007.tif" />

OCH,

<img file="LT4484B_D0008.tif" />

OH

YZ

Copulation method

<img file="LT4484B_D0009.tif" />

Y-Z

Example R

Synthesis of 3-Phenyl-1-propyl- (2S) -1- (3,3-dimethyl-1,2-dioxopentyl) -2-pyrrolidinecarboxylate (Compound of 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 resulting suspension under nitrogen for 15 min, a solution of methyl oxalyl chloride (3.20 g, 26.12 mmol) in methylene chloride (45 mL) was added dropwise, and the resulting mixture was stirred at 0 ° C for 1.5 h. water, dried over MgSO4₄ And concentrating. The crude residue was purified by passing through a silica gel column eluting with 50% ethyl acetate in hexane to give 3.52 g (88%) of product as a pink oil. Mixture of amide cis-trans rotamers; trans rotamer data are given.¹1 H NMR (CDCl 3)₃); δ, 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-d 2-dioxo-3,3-dimethylpentyl) -2-pyrrolidinecarboxylate

A solution of Metik (2S) -1- (1,2'-dioxo-2-methoxyethyl) -2-pyrrolidinecarboxylate (2.35 g, 10.90 mmol) in 30 mL of tetrahydrofuran (THF) was cooled to -78 ° C and exposed to 14, 2 ml of a 1.0 M solution of 1,1-dimethylpropylmagnesium chloride in THF, after stirring the resulting homogeneous mixture at -78 ° C for three hours, add the mixture to saturated ammonium chloride solution (100 ml) and extract with ethyl acetate. The organic phase is washed with water, dried, concentrated and the crude material obtained after solvent removal is purified by passing through a silica gel column eluted with 25% ethyl acetate in hexane. 2.10 g (75%) of oxamate are obtained as a colorless oil,¹1 H NMR (CDCl 3)₃): δ, 0.88 (t, 3H); 1.22, 1.26 (s, 3H each); 1.75 (dm, 2H), 1.87-2.10 (m, 3H); 2.23 (m, 1H); 3.54 (m, 2H); 3.76 (s, 3H); 4.52 (dm, 1H, J = 8.4, 3.4).

Synthesis of (2S) -1- (1,2-dioxo-3,3-dimethylpentyl) -2-pyrrolidinecarboxylic acid

A mixture of methyl (2S) -1- (1,2-dioxo-3,3-dimethylpentyl) -2-pyrrolidinecarboxylate (2.10 g, 8.23 ​​mmol), 1N LiOH (15 mL) and methanol (50 mL) stirred at 0 ° C for 30 min. and at room temperature overnight. The mixture was then acidified to pH 1 with 1N HCl, diluted with water and extracted with 100 mL of methylene chloride. The organic extract is washed with a saturated sodium chloride solution and concentrated to give 1.73 g (87%) of a snow-white solid which does not need to be further purified.^{1 2}H NMR (GDCi₃): δ, 0.87 (t, 3H): 1.22.1.25 (s, after 3H); 1.77 (dm, 2H); 2.02 (m, 2H); 2.17 (m, 1H); 2.25 (m, 1H); 3.53 (dd, 2H, J = 10.4, 7.3): 4.55 (dd, 1H, J = 8.6,4.1).

Synthesis of 3-Phenyl-1-propyl- (2S) -1- (3,3-dimethyl-1,2-dioxopentyl) -2-pyrrolidinecarboxylate (Compound of Example 1) (2S) -1- (1,2-Dioxo-3) , 3-Dimethylpentyl) -2-pyrrole dinecarboxylic acid (600 mg, 2.49 mmol), 3-phenyl-1-propanol (508 mg, 3.73 mmol), dicyclohexylcarbodiimide (822 mg, 3.98 mmol) , a mixture of camphor sulfonic acid (190 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 is filtered through celite to remove solids, concentrated in vacuo and the crude is purified by flash column chromatography (25% ethyl acetate in hexane). 720 mg (80%) of Example 1 is obtained, which is a colorless oil.¹1 H NMR (CDCl 3): δ, 0.84 (t, 3H); 1.19 (s, 3H); 1.23 (s, 3H); 1.70 (dm, 2H); 1.98 (m, 5H); 2.22 (m, 1H); 2.64 (m, 2H); 3.47 (m, 2H); 4.14 (m, 2H); 4.51 (d, 1H); 7.16 (m, 3H); 7.26 (m, 2H).

The method described in Example 1 was used to make the following illustrative examples:

example: 3-phenyl-1-prop-2- (E) -enyl- (2S) -1- (3,3-dimethyl-1,2-dioxopentyl) -2-pyrrolidinecarboxylate, 80%, 1 H NMR (360 MHz, CDCI₃): δ 0.86 (t, 3H); 1.21 (s, 3H); 1.25 (s, 3H); 1.54-2.10 (m, 5H); 2.10-2.37 (m, 1H);

3.52-3.55 (m, 2H); 4.56 (dd, 1H, J = 3.8, 8.9); 4.78-4.83 (m, 2H); 6.27 (m, 1H); 6.67 (dd, 1H, J = 15.9); 7.13-7.50 (m, 5H).

Example 3- (3,4,5) -trimethoxyphenyl) -1-propyl- (2S) -1- (3,3-dimethyl-1,2-dioxopentyl) -2-pyrrolidinecarboxylate, 61%, ¹1 H NMR (CDCl 3): δ 0.84 (t, 3H); 1.15 (s, 3H); 1.24 (s, 3H); 1.71 (dm, 2H); 1.98 (m, 5H); 2.24 (m, 1H); 2.63 (m, 2H); 3.51 (t, 2H); 3.79 (s, 3H); 3.83 (s, 3H); 4.14 (m, 2H); 4.52 (m, 1H); 6.36 (s, 2H).

Example 3- (3,4,5-Trimethoxyphenyl) -1-prop-2- (E) -enyl- (2S) -1- (3,3-dimethyl-1,2-dioxopentyl) -2-pyrrolidinecarboxylate, 66% , ¹1 H NMR (CDCl 3): δ 0.85 (t, 3H); 1.22 (s, 3H); 1.25 (s, 3H); 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, 1H); 4.81 (m, 2H); 6.22 (m, 1H); 6.58 (d, 1H, J = 16); 6.63 (s, 2H).

Example 3 3- (4,5) -methylenedioxyphenyl) -1-propyl (2S) -1- (3,3-dimethyl-1,2-dioxopentyl) -2-pyrrolidinecarboxylate, 82%, ¹1 H NMR (360 MHz, CDCl 3): δ 0.86 (t, 3H); 1.22 (s, 3H); 1.25 (s, 3H); 1.60-2.10 (m, 5H); 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 (m, 1H); 6.57 (dd, 1H, J = 6.2.15,8); 6.75 (d, 1H, J = 8.0); 6.83 (dd, 1H, J = 1.3, 8.0); 6.93 (s. 1H).

Example 3- (4,5) -methylenedioxyphenyl) -1-prop-2- (E) -enyl- (2S) -1 (3,3-dimethyl-1,2-dioxopentyl) -2-pyrrolidinecarboxylate, 82% , ¹1 H NMR (360 MHz, CDCl 3): δ 0.86 (t, 3H); 1.22 (s, 3H); 1.25 (s, 3H); 1.60-2.10 (m, 5H); 2.10-2.39 (m, 1H); 3.36-3.79 (m, 2H); 53 (dd, 1H, J = 3.8, 8.6); 4.61-4.89 (m, 2H); 5.96 (s, 2H); 6.10 (m, 1H); 57 (dd, 1H, J = 6.2,15.8), 6.75 (d, 1H, J = 8.0), 6.83 (dd, 1H, J = 1.3, 8.0) 6.93 (s, 1H).

Example: 3-Cyclohexyl-1-prop-2- (E) -enyl- (2S) -1 '(3,3-dimethyl-1,2-dioxopentyl) -2-pyrrolidinecarboxylate, 92%, ¹1 H NMR (360 MHz, CDCl 3)₃): δ 0.86 (t, 3H); 1.13-1.40 (m + 2 singlets, 9H total); 1.50-1.87 (m, 8H); 1.8733

2.44 (m, 6H); 3.34-3.82 (m, 2H); 4.40-4.76 (m, 3H); 5.35-5.60 (m, 1H); 5.605.82 (dd, 1H, J = 6.5.16).

Example: (1R) -1,3-Diphenyl) -1-propyl- (2S) -1- (3,3-dimethyl-1,2-dioxopentyl) -2-pyrrolidinecarboxylate, 90%, 1 H NMR (360 MHz, CDCl₃): δ 0.85 (t, 3H); 1.20 (s, 3H); 1.23 (s, 3H); 1.49-2.39 (m, 7H); 2.46-2.86 (m, 2H); 3.25-3.80 (m, 2H); 4.42-4.82 (m, 1H); 5.82 (td, 1H, J = 1,8,6,7); 7.05-7.21 (m, 3H); 7.21-7.46 (m, 7H).

Example: 3-Phenyl-1-propyl- (2S) -1- (1,2-dioxo-2- [2-furanyl]) ethyl) 2-pyrrolidinecarboxylate, 99%, ¹1 H NMR (360 MHz, CDCl 3)₃): δ 1.66-2.41 (m, 6H); 2.72 (t, 2H, J = 7.5); 3.75 (m, 2H); 4.21 (m, 2H); 4.61 (m, 1H); 6.58 (m, 1H); 7.16-7.29 (m, 5H); 7.73 (m, 2H).

Example: 3-Phenyl-1-propyl- (2S) -1- (1,2-dioxo-2- [2-thienyl]) ethyl) -2-pyrrolidinecarboxylate, 81%, 1 H NMR (300 MHz, CDCl₃): δ 1.88-2.41 (m, 6H); 2.72 (dm, 2H); 3.72 (m, 2H); 4.05 (m, 1H); 4.22 (m, 1H); 4.64 (m, 1H); 7.13-7.29 (m, 6H); 7.75 (dm, 1H); 8.05 (m, 1H).

Example: 3-Phenyl-1-propH- (2S) -1- (1,2-dioxo-2-phenyl) ethyl-2-pyrrolidinecarboxylate, 99%, 1 H NMR (300 MHz, CDCl₃): δ 1.97-2.32 (m, 6H); 2.74 (t, 2H, J = 7.5); 3.57 (m, 2H); 4.24 (m, 2H); 4.67 (m, 1H); 6.95-7.28 (m, 5H); 7.51-7.64 (m, 3H); 8.03-8.09 (m, 2H).

Example: 3- (2,5-01 Methoxyphenyl) -1-propyl- (2S) -1- (3,3-dimethyl-1,2-dioxopentyl) -2-pyrrolidinecarboxylate, 99%, 1 H NMR (300 MHz, CDCl 3) ): δ 0.87 (t, 3H); 1.22 (s, 3H); 1.26 (s, 3H); 1.69 (m, 2H); 1.96 (m, 5H); 2.24 (m, 1H); 2.68 (m, 2H); 3.55 (m, 2H); 3.75 (s, 3H); 3.77 (s, 3H); 4.17 (m, 2H); 4.53 (d, 1H); 6.72 (m, 3H).

Example: 3- (2,5-Dimethoxyphenyl) -1-prop-2- (E) -enyl- (2S) -1- (3,3-di-methyl-1,2-dioxopentyl) -2-pyrrolidinecarboxylate, 99%, 1 H NMR (300

MHz, CDCl₃): δ 0.87 (t, 3H); 1.22 (s, 3H); 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 (m, 1H); 4.81 (m, 2H); 6.29 (dt, 1H, J = 15.9); 6.98 (s, 1H).

Example 2- (3,4,5-Trimethoxyphenyl) -1-ethyl- (2S) -1- (3,3-dimethyl-1,2-dioxopentyl) -2-pyrrolidinecarboxylate, 97%, ¹1 H NMR (300 MHz, CDCl 3)₃): δ 0.84 (t, 3H); 1.15 (s, 3H); 1.24 (s, 3H); 1.71 (dm, 2H); 1.98 (m, 5H); 2.24 (m, 1H); 2.63 (m, 2H); 3.51 (t, 2H); 3.79 (s, 3H); 3.83 (S, 3H); 4.14 (m, 2H); 4.52 (m, 1H); 6.36 (s, 2H).

Example: 3- (3-Pyridyl) -1-propyl- (2S) -1- (3,3-dimethyl-1,2-dioxopentyl) -2-pyrrolidinecarboxylate, 80% ¹1 H NMR (300 MHz, CDCl 3): δ 0.85 (t, 3H); 1.23, 1.26 (S, 3H); 1.63-1.89 (m, 2H); 1.90-2.30 (m, 4H); 2.302.50 (m, 1H); 2.72 (t, 2H); 3.53 (m, 2H); 4.19 (m, 2H); 4.53 (m, 1H); 7.22 (m, 1H); 7.53 (dd, 1H); 8.45.

Example: 3 '(2'Pyridyl) -1-propyl- (2S) -1- (3,3-dimethyl-1,2-dioxopentyl) -2-pyrrolidinecarboxylate, 88%, ¹1 H NMR (300 MHz, CDCl 3)₃): δ 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, 1H); 7.09-7.19 (m, 2H); 7.59 (m, 1H);

8.53 (d, 1H, J = 4.9), Example 3- (4-Pyridyl) -1-propyl- (2S) -1- (3,3-dimethyl-1,2-dioxopentyl) -2-pyrrolidinecarboxylate , 91%, 1 H NMR (300 MHz, CDGI₃): δ 6.92-6.80 (m, 4H); 6.28 (m, 1H); 5.25 (d,. 1H, J = 5.7); 4.12 (m, 1H); 4.08 (s, 3H); 3.79 (s, 3H); 3.30 (m, 2H); 2.33 (m, 1H); 1.85-1.22 (m, 7H); 1.25 (s, 3H); 0.89 (t, 3H), J = 7.5).

Example 3 3-Phenyl-1-propyl- (2S) -1- (2-cyclohexyl-1,2-dioxoethyl) 2-pyrrolidinecarboxylate, 91%, ¹1 H NMR (300 MHz, CDCf₃): δ 1.09-1.33 (m, 5H); 1.62-2.33 (m, 12H); 2.69 (t, 2H, J = 7.5); 3.15 (dm, 1H); 3.68 (m, 2H); 4.16 (m, 2H); 4.53, 4.84 (d, 1H, total); 7.19 (m, 3H); 7.29 (m, 2H).

Example: 3-Phenyl-1-propyl-1- (2S) -1- (2-tert-butyl-1,2-dioxoethyl) -2-pyrrolidinecarboxylate, 92%, ¹1 H NMR (300 MHz, CDCl 3)₃): δ 1.29 (s, 9H); 1.94-2.03 (m, 5H); 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: 3-Phenyl'-1-propyl '(2S) -1- (2-cyclohexflethyl-1,2-dioxoethyl) -2-pyrrolidinecarboxylate, 97%,'¹1 H NMR (300 MHz, CDCl 3)₃): δ 0.88 (m, 2H); 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 (d, 1H); 7.177.32 (m, 5H).

Example: 3- (3-Pyridyl) -1-propyl- (2S) -1- (2-cyclohexylethyl-1,2-dioxoethyl) -2-pyrrolidinecarboxylate, 70% ¹1 H NMR (300 MHz, CDCl 3)₃): δ 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: 3- (3-Pyridyl) -1-propyl- (2S) -1- (2-tert-butylethyl-1,2-dioxoethyl) -2-pyrrolidinecarboxylate, 83%, ¹1 H NMR (300 MHz, CDCl 3)₃): δ 1.29 (3, 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, 2H); 4.52 (m, 1H); 7.19-7.25 (m, 1H); 7.53 (m, 1H); 8.46 (m, 2H).

Example: 3,3-Diphenyl-1-propyl- (2S) -1- (3,3-dimethyl-1,2-dioxopentyl) -2-pyrrolidinecarboxylate, 99%, ¹1 H NMR (300 MHz, CDCl 3)₃): δ ♦ 0.85 (t, 3H); 1.21, 1.26 (s, 3H each); 1.68-2.04 (m, 5H); 2.31 (m, 1H); 2.40 (m, 2H); 3.51 (m, 2H); 4.08 (m, 3H); 4.52 (m, 1H); 7.18-7.31 (m, 10H).

Example: 3- (3-Pyridyl) - 't -propyl- (2S) -1- (2-cyclohexyl-1)_!2-dioxoethyl) -2-pyrrolidinecarboxylate, 88%, ¹1 H NMR (300 MHz, CDCl 3)₃): δ 1.24-1.28 (m, 5H); 1.88-2.35 (m, 11H); 2.72 (t, 2H, J = 7.5); 3.00-3.33 (dm,

1H); 3.69 (m, 2H); 4.19 (m, 2H): 4.55 (m, 1H); 7.20 * 7.24 (m, 1H); 7.53 (m, 1H); 8.47 (m, 2H).

Example 3- (3-Pyridyl) -1-propyl- (2S) -N- (1,2-thienyl] glyoxyl) pyrrolidinecarboxylate, 49%, 1 H NMR (300 MHz, CDCl 3): δ 1.81-2.39 (m, 6H); 2.72 (dm, 2H); 3.73 (m, 2H); 4.21 (m, 2H); 4.95 (m, 1H); 7.19 (m, 2H); 7.61 (m, 1H); 7.80 (d, 1H); 8.04 (d, 1H); 8.46 (m, 2H).

Example: 3,3'-Diphenyl-1-propyl- (2S) -1- (3,3-dimethyl-1,2-dioxobutyl) -2-pyrrolidinecarboxylate, 99%, 1 H NMR (300 MHz, CDCl 3): δ

1.27 (s, 9H); 1.96 (m, 2H); 2.44 (m, 4H); 3.49 (m, 1H); 3.64 (m, 1H); 4.08 (m, 4H): 4.53 (dd, 1H); 7.24 (m, 10H).

example: 3,3-DWenyl-1-propyl- (2S) '1-cyclohexylglyoxyl-2-pyrrolidinecarboxylate, 91%, 1 H NMR (300 MHz, CDCl 3): δ 1.32 (m, 6H); 1.54-2.41 (m, 10H); 3.20 (dm, 1H); 3.69 (m, 2H); 4.12 (m, 4H); 4.52 (d, 1H);

7.28 (m, 10H), example:. 3,3-Diphenyl-1-propyl- (2S) -1- (2-thienyl) glyoxyl-2-pyrrolidinecarboxylate, 75%, 1 H NMR (300 MHz, CDCl₃): δ 2.04 (m, 3H); 2.26 (m, 2H); 2.48 (m, 1H); 3.70 (m, 2H); 3.82-4.18 (m, total 3H); 4.64 (m, 1H); 7.25 (m, 11H); 7.76 (dd, 1H); 8.03 (m, 1H).

The requisite synthesized alcohols can be prepared by a variety of methods known to those skilled in the art of organic synthesis. As shown in Scheme II, the alkyl or aryl aldehydes can be homologated to the phenitopropanols by the action of methyl (triphenylphosphoranitidene) acetate to form various trans-cinamates; the latter may be reduced to the saturated alcohols by treatment with excess lithium aluminum hydride, or by gradual reduction of the double bond by catalytic hydrogenation and reduction of the resulting saturated ester with appropriate reducing agents. Alternatively, trans-cinnamates may be reduced to (E) -alyl alcohols using diisobutylaluminum hydride.

R-CHO

Ph₃P = CHCOOCH₃THF

Diisobutylaluminium / hydride

Scheme H

Lithium aluminum hydride

<img file="LT4484B_D0010.tif" />

COOCH,

<img file="LT4484B_D0011.tif" />

Lithium aluminum hydride or

Diisobutylaluminium hydride

R

<img file="LT4484B_D0012.tif" />

/ COOCH₃

Longer chain alcohols can be prepared by homologating benzyl or higher aldehydes. Alternatively, these aldehydes may be prepared from the corresponding phenylacetic and higher acids, phenethyl and higher alcohols.

General procedure for the synthesis of acrylic esters (example of methyl (3.4.5-trimethoxy) -transcinamate)

A solution of 3,4,5-trimethoxybenzaldehyde (5.0 g, 25.48 mmol) and methyl (triphenylphosphoranylidene) acetate (10.0 g, 29.91 mmol) in tetrahydrofuran (250 mL) was refluxed overnight. 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 give 5.63 g (88%) of cinnamate as a white crystalline solid; ¹1 H NMR (300 MHz, CDCl 3)₃): δ 3.78 (s, 3H); 3.85 (s, 6H); 6.32 (d, 1H, J = 16); 6.72 (s, 2H); 7.59 (d, 1H, J = 16).

General procedure for the synthesis of saturated alcohol from acrylic esters (example of (3,4,5-trimethoxy) phenylpropanol)

A solution of methy- (3,4,5-trimethoxy) -trans-cinnamate (1.81 g, 7.17 mmol) in tetrahydrofuran (30 mL) was added dropwise to a solution of lithium aluminum hydride (14 mmol) in THF (35 mL) under a nitrogen atmosphere. . After the addition is complete, the mixture is heated to 75 ° C for 4 hours. After cooling, the reaction was quenched by 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 solid collected on the filter was washed with ethyl acetate. The combined organic fractions were washed with water, dried, concentrated in vacuo and the residue purified by passing through a silica gel column eluted with ethyl acetate. 0.86 g (53%) of alcohol is obtained which is a clear oil;¹1 H NMR (300 MHz, CDCl 3): δ 1.23 (pi, 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 allyl alcohol from acrylic esters (Example of (3,4,5-trimethoxy) phenylprop-2- (E) -enol)

A solution of methyl- (3,4,5-trimethoxy) -transcinnamate (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 1). , 0 M solution; 11.25 mmol). The reaction mixture is stirred for 3 hours. At 0 ° C, the reaction was quenched with 3 mL of methanol and then 1Ν HCl to pH 1. The reaction mixture was extracted with ethyl acetate, the organic phase was washed with water, dried and concentrated. Purification by passing through a silica gel column eluting with 25% ethyl acetate in hexane afforded 0.96 g (80%) of a viscous oil; ¹1 H NMR (360 MHz, CDCl 3)₃): δ 3.78 (s, 3H); 3.87 (s, 6H); 4.32 (d, 2H, J = 5.6); 6.29 (dt, 1H, J = 15.8, 5.7); 6.54 (d, 1H, J = 15.8); 6.61 (s, 2H).

Having described the invention in this way, it is understood that the same subject matter may have many variations. Such variations are not to be construed as departures from the spirit and scope of the invention, and it is to be understood that all such modifications are intended to be covered by the following claims.