RU2269514C9 - Small molecules as rotamase enzyme activity inhibitors - Google Patents

Abstract

FIELD: biochemistry.

SUBSTANCE: invention relates to new compounds of formula

wherein R₁ represents linear or branched C₁-C₉-alkyl optionally substituted with C₃-C₈-cycloalkyl, C₆-cycloalkyl, 2-furil; 3-furil, 2-thiazolyl, 2-thenyl, 3-thienyl, phenyl; X represents oxygen, NR₄, wherein R₄ is H, C₁-C₄-alkyl; Z represents H with the proviso, that when X and Y are oxygen, R₁ is not methyl, ethyl, isopropyl, isobutyl or phenyl; and when X is oxygen, and Y is NR₂, wherein R₂ is hydrogen, methyl, isopropyl or tert-butyl R₁ is not methyl. Compounds of present invention are useful as synthetic intermediates for bioactive substances.

EFFECT: new synthetic intermediates for bioactive substances.

8 cl, 28 dwg, 3 tbl, 38 ex

Description

BACKGROUND

1. Field of invention

The present invention relates to neurotrophic compounds having an affinity for FKBP-type immunophilins, their preparation, as well as their use as inhibitors of the enzymatic activity associated with immunophilin proteins, in particular as inhibitors of the enzymatic activity of peptidyl prolyl isomerase or rotamase.

2. Description of previous achievements in this field

The term "immunophilin" refers to a number of proteins that serve as receptors for essential immunosuppressive drugs, such as cyclosporin A (CsA), FK506 and rapamycin. Known classes of immunophilins are cyclophilins and FK506 binding proteins, such as FKBP. While cyclosporin A binds to cyclophilin, FK506 and rapamycin bind to FKBP. These complexes formed by immunophilin and a drug are involved in a number of intracellular signal transduction systems, especially in the immune and nervous systems.

Immunophilins are known to have peptidyl-prolyl-isomerase (PPI) or rotamase enzymatic activity. It was shown that rotamase activity is involved in the catalysis of the mutual conversion of cis and trans isomers of immunophilin proteins.

Immunophilins were first discovered and studied in immune tissue. Initially, experts assumed that the inhibition of the rotamase activity of immunophilins leads to suppression of T-cell proliferation, which in turn causes the immunosuppressive effect inherent in immunosuppressive drugs such as cyclosporin A, FKBP and rapamycin. As further studies have shown, inhibition of rotamase activity alone is not sufficient for immunosuppressive activity (Schreiber et al., Science, 1990, vol. 250, pp. 566-559). It has been demonstrated that the mode of action of the aforementioned immunophilin and drug complexes is due to their interaction with triple target proteins (Schreiber et al., Cell, 1991, vol. 66, pp. 807-815). The complexes formed by the drug and immunophilin (such as FKBP-FK506 and FKBP-CsA) bind to the calcineurin enzyme, inhibiting T cell receptor signaling, which leads to T cell proliferation. The complex formed by rapamycin and FKBP interacts similarly with the RAFT1 / FRAP protein and inhibits signal transmission from the IL-2 receptor.

It was found that immunophilins are present in high concentrations in the central nervous system, where their number is 10-50 times greater than in the immune system. As it turned out, in nerve tissues, immunophilins have an effect on the growth of a neuron, the synthesis of nitric oxide, and the secretion of neurotransmitters.

Picomolar concentrations of immunosuppressants, such as FK506 and rapamycin, stimulate external axon growth in PC12 and sensory nerve cells, in particular in spinal cell nodes (DRGs) (Lyons et al., Proc. Of Natl. Acad. Sci., 1994, vol. 91, pp. 3191-3195). In experiments on whole animals, it was found that FK506 stimulates nerve regeneration after facial nerve surgery and causes functional recovery in animals with sciatic nerve damage.

It was surprising that drugs with a high affinity for FKBP are potent rotamase inhibitors with neurotrophic effects (Lyons et al.). Based on these findings, it was proposed to use immunosuppressants in the treatment of various peripheral neuropathies, as well as to enhance the recovery of nerves in the central nervous system (CNS). Studies have shown that the cause of neurodegenerative disorders such as Alzheimer's disease, Parkinson's disease and amyotrophic lateral sclerosis (ALS) can be the loss or decrease in the availability of neurotrophic substances specific to a specific population of neurons affected by this disease.

Several neurotrophic factors affecting certain populations of neurons in the central nervous system have been identified. In particular, it has been suggested that Alzheimer's disease results from a decrease in the content or disappearance of nerve growth factor (NGF). In this regard, it has been suggested that Alzheimer's disease can be treated with exogenous nerve growth factor or other neurotrophic proteins such as cerebral nerve factor (BDNF), glial nerve factor, ciliary neurotrophic factor, and neurotropin-3, as a result of which it will be observed increased survival of a degenerating population of neurons.

The clinical use of these proteins for the treatment of various neurological diseases is complicated by the complexity of the introduction and bioavailability of large proteins in relation to the targets of the nervous system. In contrast, immunosuppressive drugs with neurotrophic activity are characterized by relatively small sizes, as well as excellent bioavailability and specificity. However, in the chronic use of immunosuppressants, a number of potentially serious side effects are observed, including neurotoxicity (for example, impaired glomerular filtration and irreversible interstitial fibrosis (Corr et al., 1991, J. Am. Soc. Nephrol. 1: 162); neurological deficits (e.g. involuntary tremors) or nonspecific cerebral angina (e.g. non-localizable headaches (De Groen et al., 1987, Engl. J. Med. 317: 861)); as well as vascular hypertension with its complications (Kahan et al., 1989, N. Engl. J. Med. 321: 1725).

In order to avoid these side effects associated with the use of immunosuppressive compounds, the present invention provides the use of compounds that are not immunosuppressive and contain FKBP rotamase inhibitors in the form of small molecules, which serve to stimulate the growth and regeneration of neurons in various neuropathological conditions, which can be alleviated restoration of nerves, in particular with damage to peripheral nerves as a result of physical trauma or painful condition I, such as diabetes, physical damage to the central nervous system (spinal or brain), brain damage during shaking, as well as for the treatment of neurological diseases accompanied by neurodegeneration, including Parkinson's disease, Alzheimer's disease and amyotrophic lateral sclerosis.

SUMMARY OF THE INVENTION

The present invention relates to a new class of neurotrophic compounds having an affinity for FKBP-type immunophilins. Associated with these proteins, the considered neurotrophic compounds are potent inhibitors of the enzymatic activity associated with immunophilin proteins, specifically, the enzymatic activity of rotamase, as a result of which stimulation of regeneration and lengthening of neurons occurs. A key distinguishing feature of the compounds of the present invention is their lack of any significant immunosuppressive activity in addition to their neurotrophic activity.

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

in which R ₁ represents a group selected from linear or branched C ₁ -C ₉ alkyl or alkenyl groups optionally substituted with C ₃ -C ₈ cycloalkyl, C ₃ or C ₅ cycloalkyl, C ₅ -C ₇ cycloalkenyl or an Ar ₁ radical wherein said alkyl, alkenyl, cycloalkyl or cycloalkenyl groups may optionally be substituted with C ₁ -C ₄ alkyl, C ₁ -C ₄ alkenyl or hydroxyl, and Ar ₁ is a radical selected from 1-naphthyl, 2-naphthyl, 2 -indolyl, 3-indolyl, 2-furyl, 3-furyl, 2-thiazolyl, 2-thienyl, 3-thienyl, 2-, 3- or 4-pyridine a, and phenyl, having one to three substituents which are independently selected from the following: hydrogen, halogen, hydroxyl, nitro, trifluoromethyl, linear or branched C ₁ -C ₆ alkyl or alkenyl, C ₁ -C ₄ alkoxy or C ₁ -C ₄ alkenyloxy, phenoxy group, benzyloxy group, as well as an amino group;

X corresponds to a group selected from the following: oxygen, sulfur, methylene (CH ₂ ) or H ₂ ;

Y denotes a group representing oxygen or NR ₂ where R ₂ is hydrogen or a linear or branched C ₁ -C ₆ alkyl; but

Z is a residue corresponding to linear or branched C ₁ -C ₆ alkyl or alkenyl;

wherein said alkyl chain is substituted at one or more of the positions Ar ₁ as defined above; C ₃ -C ₈ cycloalkyl; cycloalkyl attached via a straight or branched C ₁ -C ₆ alkyl or alkenyl chain; or Ar ₂ , which is a group selected from the following: 2-indolyl, 3-indolyl, 2-furyl, 3-furyl, 2-thiazolyl, 2-thienyl, 3-thienyl, 2-, 3- or 4- pyridyl, as well as phenyl, having one to three substituents, which are independently selected from the following: hydrogen, halogen, hydroxyl, nitro group, trifluoromethyl, linear or branched C ₁ -C ₆ alkyl or alkenyl, C ₁ -C ₄ alkoxyl or C ₁ -C ₄ alkenyloxy, phenoxy group, benzyloxy group, as well as an amino group; but

Z may also be represented by a fragment

where R ₃ selected from the group consisting of linear or branched alkyl C _1-8 , possibly substituted by C ₃ -C ₈ cycloalkyl or the above Ar ₁ , or unsubstituted Ar ₁ ;

X ₂ is oxygen or NR ₅ , where R _{5 is} selected from the group consisting of hydrogen, C ₁ -C ₆ straight or branched alkyl or alkenyl; but

R _{4 is} selected from the group consisting of phenyl, benzyl, linear or branched C ₁ -C ₅ alkyl or alkenyl, and also linear or branched C ₁ -C ₅ alkyl or alkenyl substituted with phenyl; and its pharmaceutically acceptable salts or hydrates.

Another preferred embodiment of the present invention is a neurotrophic compound according to the formula

wherein

R ₁ represents a C _1-9 linear or branched alkyl or alkenyl group optionally substituted with a C ₃ -C ₈ cycloalkyl, C ₃ - or C ₅ cycloalkyl, C ₅ -C ₇ cycloalkenyl or Ar ₁ , wherein said alkyl, alkenyl, cycloalkyl or cycloalkenyl groups may be optionally substituted by C ₁ - ₄ alkyl, C ₁ -C ₄ alkenyl, or hydroxy, and where Ar ₁ is selected from the group consisting of 1-napthyl, 2-napthyl, 2-indolyl, 3-indolyl, 2-furyl , 3-furyl, 2-thiazolyl, 2-thienyl, 3-thienyl, 2-, 3- or 4-pyridyl or phenyl bearing from one to three substituents which are independently selected from the following: hydrogen, halogen, hydroxyl, nitro group, trifluoromethyl, linear or branched C ₁ -C ₆ alkyl or alkenyl, C ₁ -C ₄ alkoxyl or C ₁ -C ₄ alkenyloxy, phenoxy group, benzyloxy group, as well as an amino group; but

Z is C _2-6 linear or branched alkyl or alkenyl; wherein said alkyl chain is substituted at one or more positions as defined above Ar ₁ ; C ₃ -C ₈ cycloalkyl; cycloalkyl attached via a straight or branched C ₁ -C ₆ alkyl or alkenyl chain; or Ar ₂ , and Ar ₂ 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, or phenyl having from one to three substituents which are independently selected from the following: hydrogen, halogen, hydroxyl, nitro group, trifluoromethyl, linear or branched C ₁ -C ₆ alkyl or alkenyl, C ₁ -C ₄ alkoxyl or C ₁ -C ₄ alkenyloxy, phenoxy group, benzyloxy group, as well as an amino group; or its pharmaceutically acceptable salts or hydrates.

Another preferred embodiment of the present invention is a neurotrophic compound having an affinity for FKBP type immunophilins that inhibit the rotamase activity of said immunophilin.

Another preferred embodiment of the present invention is a method of treating a neurological disease in an animal, comprising using a therapeutically effective amount of a compound having an affinity for immunophilins such as FKBP, which inhibits the rotamase activity of said immunophilin.

Another preferred embodiment of the present invention is a method of activating the regeneration and growth of neurons in mammals, comprising exposing the mammal to an effective amount of a neurotrophic compound having an affinity for immunophilins such as FKBP, which inhibits the rotamase activity of said immunophilin.

Another preferred embodiment of the present invention is a method of preventing nerve degradation in animals, comprising exposing the animal to an effective amount of a neurotrophic compound having an affinity for immunophilins such as FKBP, which inhibits the rotamase activity of said immunophilin.

Another preferred embodiment of the present invention is a neurotrophic compound, which is an N-glyoxyl-prolyl ether of the formula:

wherein

R ₁ represents a linear or branched C ₁ -C ₅ alkyl or alkenyl group optionally substituted with a C ₃ -C ₆ cycloalkyl, or Ar ₁ , where Ar _{1 is} selected from the group consisting of 2-furyl, 2-thienyl or phenyl;

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

Y is oxygen; but

Z is a linear or branched chain alkyl or alkenyl, wherein said alkyl chain is substituted at one or more of the positions of Ar ₁ described above, C ₃ -C ₆ cycloalkyl or Ar ₂ which is selected from the group consisting of 2-, 3- or 4-pyridyl or phenyl having from one to three substituents independently selected from hydrogen or C ₁ -C ₄ alkoxyl.

The most preferred neurotrophic compounds representing N-glyoxyl-prolyl ethers corresponding to the above formula are the following:

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-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-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) -N - ([2-thienyl] glyoxyl) pyrrolidinecarboxylate.

BRIEF DESCRIPTION OF ILLUSTRATIONS

Figure 1, is a micrograph of the chicken spinal node treated with various concentrations of the compound described in example 17. As follows from Figure 1, the compound corresponding to example 17 of the present invention has a powerful activating effect on axon extension in sensory neuron cultures. Explant cultures isolated on day 9-10 of chicken embryonic development from the spinal node were treated with various concentrations of the compound of Example 17. After 48 hours, the number of axons was counted, the length of which exceeded the corresponding value for the initial DRG explant. From the indicated value, the number of similar axons in explants not treated with the compound from Example 17 was subtracted, resulting in an indicator characterizing the stimulation of axon elongation under the action of the compound from Example 17. Micrographs of DRG's treated with the compound from Example 17 are presented, as well as a quantitative dose-dependent elongation of the axons eliminated by the compound of example 17.

Figure 2 graphically shows the quantitative indicators of axon elongation in the chicken spinal nodes treated with various concentrations of the compound of Example 17. As follows from figure 2, the compound corresponding to example 17 of the present invention has a powerful activating effect on axon extension in cultures of sensitive neurons. Explant cultures isolated on day 9-10 of chicken embryonic development from the spinal nodes were treated with various concentrations of the compound of Example 17. After 48 hours, the number of axons whose length exceeded the corresponding value for the initial DRG explant was counted. From the indicated value, the number of similar axons in the explants not treated with the compound from Example 17 was subtracted, as a result of which an indicator was obtained characterizing the stimulation of axon extension under the action of the compound from Example 17. The dose-effect quantitative dependences of axon extension under the action of the compound from Example 17 are presented. .

Figure 3 is a micrograph of sections of the sciatic nerve of a rat. As follows from figure 3, the compound described in example 1 of the present invention activates the regeneration of neurons after damage to the sciatic nerve. In male Sprague-Dawley rats weighing 150 g, sciatic nerves were squeezed at the level of the thighs. Over the next 21st day, the indicated rats were administered once daily subcutaneously with the compound of Example 1 (30 mg / kg), an inactive compound (30 mg / kg), or a lipid carrier. Then the animals were sacrificed, sciatic nerves were isolated, nerve sections were obtained at a site 2 mm distal to the lesion and stained with Holmes silver dye (to count the number of axons), as well as Luxol fast blue (to detect remyelination). This micrograph (magnification 630x) shows sections of the sciatic nerve in newly operated rats, as well as animals treated with a vehicle, a compound from Example 1, or an inactive compound; 4 animals per group.

Figure 4 graphically shows the [ ³ H] -CFT binding per μg of striatal membrane protein. As can be seen from figure 4, the neuroimmunophilin ligands provided by the present invention activate the recovery of mouse dopamine neurons after treatment with MPTP. CD1 mice weighing 25 g were injected intraperitoneally with 30 mg / kg MPTP daily for 5 days. In addition, subcutaneous administration of a lipid carrier, a compound from Example 1 (100 mg / kg), or a compound from Example 17 (40, 20 or 10 mg / kg) was carried out subcutaneously daily for the animals in question for the entire duration of the MPTP application and for the next 5 days . After another 18 days, the mice were sacrificed, striped bodies isolated from 5 animals of each group were combined, and then they were used to obtain preparations of washed membranes. In order to determine the number of dopamine transporters on living nerve endings, the binding rate of [ ³ H] -CFT with striatal membrane preparations obtained from various animal groups was evaluated. Binding intensity in the presence of 10 μM unlabeled CFT was an indicator of nonspecific binding; this value was subtracted from the indicator of the total binding intensity, resulting in a characteristic of specific [ ³ H] -CFT binding. The binding intensity was an indicator of the amount of the corresponding protein in the striatal membranes of each of the experimental groups. In order to obtain data on the amount of tyrosine hydrolase (TN) in the striatum, in axon bundles of the middle part of the forebrain, as well as on the nigral level of the enzyme (which is a characteristic of functional dopaminergic neurons), coronal and sagittal sections of the brain of animals treated with either MPTP, or a drug, was stained with anti-tyrosine hydrolase lg.

Figure 5 shows a histogram for [ ³ H] -CFT constructed for 200 μg of membrane protein. As can be seen from figure 5, the neuroimmunophilin ligands provided by the present invention, activate the recovery of dopamine neurons after processing mice with MRTP in accordance with the procedure described for figure 4.

6 is a micrograph (magnification 630x) of coronary and sagittal sections of the brain. Figure 6 shows sections of the brain stained with anti-tyrosine hydroxylase lg; while the experimental animals were treated with either MPTP or a drug; this treatment allows us to estimate the amount of TN in the striatum, which is a characteristic of functional dopaminergic neurons.

Fig.7 is a micrograph (magnification 50x) of the coronary and sagittal sections of the brain. Figure 7 shows sections of the brain stained with anti-tyrosine hydroxylase lg; while the experimental animals were treated with either MPTP or a drug; this treatment allows you to evaluate the nigral levels of TN, which is a characteristic of functional dopaminergic neurons.

Fig. 8 is a photomicrograph (400x magnification) of coronary and sagittal sections of the brain. Fig. 8 shows sections of the brain stained with anti-tyrosine hydroxylase lg; while the experimental animals were treated with either MPTP or a drug; this treatment allows us to estimate the amount of TN in the axon bundles of the middle part of the forebrain, which is a characteristic of functional dopaminergic neurons.

DETAILED DESCRIPTION OF THE INVENTION

The new neurotrophic compounds of the present invention are characterized by a relatively small molecular size compared to other known compounds, such as rapamycin, FK506 and cyclosporin, which are capable of binding to immunophilins of the FKBP type.

The neurotrophic compounds of the present invention have an affinity for FK-506 binding proteins, such as FKBP-12. Surprisingly, when binding to FKBP, the neurotrophic compounds of the present invention inhibit the prolyl-peptidyl cis-trans isomerase or rotamase activity of the binding protein and stimulate axon growth in the absence of an immunosuppressive effect.

More specifically, the present invention relates to a new class of neurotrophic compounds corresponding to the formula

in which R ₁ represents a linear or branched C ₁ -C ₉ alkyl or alkenyl group optionally substituted with a C ₃ -C ₈ cycloalkyl, C ₃ or C ₅ cycloalkyl, C ₅ -C ₇ cycloalkenyl or an Ar ₁ radical, wherein said alkyl, alkenyl, cycloalkyl or cycloalkenyl groups may optionally be substituted with C ₁ -C ₄ alkyl, C ₁ -C ₄ alkenyl or hydroxyl, and Ar _{1 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, as well as phenyl, bearing from one three substituents that are independently selected from the following: hydrogen, halogen, hydroxyl, nitro group, trifluoromethyl, linear or branched C ₁ -C ₆ alkyl or alkenyl, C ₁ -C ₄ alkoxyl or C ₁ -C ₄ alkenyloxy, phenoxy group, benzyloxy group as well as an amino group;

X is oxygen, sulfur, methylene (CH ₂ ) or H ₂ ;

Y is oxygen or NR ₂ where R ₂ is hydrogen or C ₁ -C ₆ alkyl; but

Z is a linear or branched C ₂ -C ₆ alkyl or alkenyl; moreover, the alkyl chain is substituted at one or more positions by the radical Ar ₁ as defined above; C ₃ -C ₈ cycloalkyl; cycloalkyl connected by a linear or unbranched C ₁ -C ₆ alkyl or alkenyl chain; or an Ar ₁ radical 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, as well as phenyl bearing from one to three substituents, which are independently selected from the group consisting of hydrogen, halogen, hydroxyl, nitro group, trifluoromethyl, linear or branched C ₁ -C ₆ alkyl or alkenyl, C ₁ -C ₄ alkoxyl or C ₁ —C ₄ alkenyloxy, phenoxy groups, benzyloxy groups, and also amino groups; but

Z may also be represented by a fragment

where R ₃ selected from the group consisting of linear or branched C ₁ -C ₈ alkyl, possibly substituted with C ₃ -C ₈ cycloalkyl or the above radical Ar ₁ or unsubstituted Ar ₁ ;

X ₂ is oxygen or NR ₅ , where R _{5 is} selected from the group consisting of hydrogen, linear or branched C ₁ -C ₆ alkyl or alkenyl; but

R _{4 is} selected from the group consisting of phenyl, benzyl, linear or branched C ₁ -C ₅ alkyl or alkenyl, and also linear or branched C ₁ -C ₅ alkyl or alkenyl substituted with phenyl;

as well as pharmaceutically acceptable salts of these compounds and their hydrates.

Preferred are compounds corresponding to the formula:

in which R ₁ represents a C ₁ -C ₉ straight or branched chain alkyl or alkenyl group optionally substituted with C ₃ -C ₈ cycloalkyl, C ₃ or C ₅ cycloalkyl, C ₅ -C ₇ cycloalkenyl or an Ar ₁ radical, wherein the alkyl, alkenyl, cycloalkyl or cycloalkenyl groups may optionally be substituted with C ₁ -C ₄ alkyl, C ₁ -C ₄ alkenyl or hydroxyl, and Ar _{1 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, as well as phenyl bearing from about Foot to three substituents which are independently selected from hydrogen, halogen, hydroxyl, nitro, trifluoromethyl, linear or branched C ₁ -C ₆ alkyl or alkenyl, C ₁ -C ₄ alkoxy or C ₁ -C ₄ alkenyloxy, phenoxy, benzyloxy, and also amino groups; but

Z is a linear or branched C ₂ -C ₆ alkyl or alkenyl; wherein said alkyl chain is substituted at one or more positions with the radical Ar ₁ described above; C ₃ -C ₈ cycloalkyl; cycloalkyl connected by a linear or unbranched C ₁ -C ₆ alkyl or alkenyl chain; or an Ar ₂ radical 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, as well as

phenyl bearing from one to three substituents that are independently selected from the group consisting of hydrogen, halogen, hydroxyl, nitro group, trifluoromethyl, linear or branched C ₁ -C ₆ alkyl or alkenyl, C ₁ -C ₄ alkoxyl or C ₁ -C _4- alkenyloxy, phenoxy groups, benzyloxy groups, as well as amino groups;

or pharmaceutically acceptable salts of these compounds and their hydrates.

Preferred neurotrophic compounds, which are N-glyoxyl prolyl ether, correspond to the formula:

in which R ₁ represents a linear or branched C ₁ -C ₅ alkyl or alkenyl group optionally substituted with a C ₃ -C ₆ cycloalkyl or Ar ₁ radical, where Ar _{1 is} selected from the group consisting of 2-furyl, 2-thienyl or phenyl;

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

Y is oxygen; but

Z is linear or branched alkyl or alkenyl; moreover, the indicated alkyl chain is substituted at one or more positions by the above radical Ar ₁ , C ₃ -C ₆ cycloalkyl or radical Ar ₂ which is selected from the group consisting of 2-, 3- or 4-pyridyl, or phenyl bearing from one to three substituents independently selected from the group consisting of hydrogen or C ₁ -C ₄ alkoxyl.

Compounds envisioned by the present invention exist in the form of stereoisomers or enantiomers or diastereoisomers. The stereochemistry at position 1 (see formula 1) is R or S, however, the S configuration is preferred. The scope of the present invention covers enantiomers, racemic form, as well as mixtures of diastereoisomers. The separation of enantiomers and diastereoisomers can be carried out using methods well known in the art.

Immunophilins, such as FKBP, are known to preferably recognize peptide substrates containing Haa-Progo-Yaa motifs, where Haa 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). Compounds modified in this way, which are prolyl-containing pseudo-symmetric peptides bearing lipophilic substituents, must bind to the hydrophobic backbone of the active site of FKBP with high efficiency and inhibit its rotamase activity.

The structure of the preferred compounds provided by the present invention includes R ₁ groups that are not stereochemically bulky compared to the known shape and size of the hydrophobic backbone in the active site of the FKBP molecule. Accordingly, very large and / or highly substituted R ₁ groups must bind to the FKBP active site with less efficiency.

Among the preferred compounds provided by the present invention include:

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-dioxopentyl) -2-pyrrolidinecarboxylate,

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

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

(1R) -1,3-diphenyl-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 2-pyrrolidinecarboxylate,

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

3- (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-pyrrolidinecarboxylate,

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

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

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

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

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

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

3,3-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,

3,3-diphenyl-1-propyl (2S) -1- (2-thienyl) glyoxyl-2-pyrrolidinecarboxylate.

The most preferred neurotrophic compounds, representing N-glyoxyl-prolyl esters, are among the following:

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-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-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-cyclohexylhydroxyl-2-pyrrolidinecarboxylate, and

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

Compounds provided by the present invention can be used in the form of salts formed by inorganic or organic acids and bases. Among the salts formed by acids are the following: acetate, adipate, alginate, aspartate, benzoate, benzenesulfonate, bisulfate, butyrate, citrate, camphor, camphorsulfonate, cyclopentane propionate, digluconate, dodecyl sulfate, heptane sulfate, hepta sulfate, fumarate, , hexanoate, hydrochloride, hydrobromide, hydroiodide, 2-hydroxyethanesulfonate, lactate, maleate, methanesulfonate, 2-naphthyl sulfonate, nicotinate, oxalate, pamoate, pectinate, propionate, succinate, tartrate, thiocyanate, and tosylate, and al. Salts formed by bases include: ammonium salts, alkali metal salts (such as sodium and potassium salts), alkaline earth metal salts (such as calcium and magnesium salts), salts formed by organic bases (such as dicyclohexylamine salts and N -methyl-D-glucamine), as well as salts formed by amino acids (such as arginine, lysine, etc.). In addition, basic nitrogen-containing groups can be quaternized with agents such as lower alkyl halides (e.g. methyl, ethyl, propyl and pentyl chlorides, bromides or iodides), dialkyl sulfates (e.g. dimethyl, diethyl , dibutyl and diamyl sulfates), long chain hydrocarbon halides (e.g. decyl, lauryl, myristyl and stearyl chlorides, bromides or iodides), aralkyl halides (e.g. benzyl and phenylethyl bromides), as well as other compounds. This produces products that are soluble or dispersible in water or oil.

The neurotrophic compounds provided by the present invention can be periodically applied to a patient undergoing treatment for neurological disorders, or for other purposes when it is desirable to stimulate the regeneration and growth of nerves, for example, with various peripheral neuropathic and neurological disorders accompanied by neurodegeneration. In addition, the compounds provided by the present invention can be used for animals other than humans in the treatment of a variety of neurological disorders of mammals.

The novel compounds provided by the present invention are potent inhibitors of rotamase activity and have an excellent neurological effect. The indicated effect can be used to stimulate damaged neurons, activate neuron regeneration, prevent neurodegradation, and also in the treatment of certain neurological disorders, which are known to be accompanied by neuronal degradation and peripheral neuropathies. These neurological disorders that can be treated as described above include, but are not limited to, trigeminal neuralgia, laryngopharyngeal neuralgia, severe pseudoparalytic myasthenia gravis, Bell's paralysis, muscular dystrophy, amyotrophic lateral sclerosis, progressive muscular atrophy, progressive bulbar hereditary muscle atrophy, syndromes caused by protrusion, rupture or prolapse of the intervertebral disc, cervical spondylosis, plexus disease, syndrome end-to-end chest injuries, peripheral neuropathy (caused, in particular, by lead poisoning, dapsone, tick bites), porphyria, or Guillain-Barre-Strol syndrome (Gullain-Barre), Alzheimer's disease, Parkinson's disease, and also other diseases.

For these purposes, the compounds provided by the present invention can be administered orally, parenterally, by inhalation, topically, rectally, nasally, buccally, vaginally, or using an implantable container using dosage formulations containing conventional non-toxic pharmaceutically acceptable carriers, adjuvants and excipients. In the present application, the term “parenteral administration” includes subcutaneous, intravenous, intramuscular, intraperitoneal, intracapsular, intraventricular, intrathoracic or intracranial administration by injection or infusion.

In order for the complex formed by immunophilin and the drug compound to be therapeutically effective against targets of the central nervous system, it must successfully overcome the blood-brain barrier in the case of peripheral administration. Compounds provided by the present invention that are not able to cross the blood-brain barrier can be successfully applied by intraventricular administration.

The pharmaceutical compositions contemplated may be in the form of a sterile preparation for injection, for example, in the form of a sterile aqueous or oily suspension for injection. Said suspension may be prepared according to methods well known in the art using suitable dispersing or wetting agents as well as suspending agents. In addition, the injectable sterile preparation may be an injectable sterile solution or suspension in a non-toxic diluent or solvent suitable for parenteral administration (for example, a solution in 1,3-butanediol). Suitable carriers and solvents suitable for this use include water, Ringer's solution, and isotonic sodium chloride solution. In addition, sterile, stable oils are conventionally used as a solvent or suspension medium. For this purpose any stable soft oil, including artificial mono- or diglycerides, can be used. Fatty acids, such as butyric acid and its glyceride derivatives, as well as olive and castor oils (especially in their polyoxyethylated forms), are used in the manufacture of injectable preparations. In addition, these oil solutions or suspensions may contain long hydrocarbon chain alcohols as diluents or dispersants.

These compounds can be administered orally, for example, in the form of capsules or tablets, or in the form of an aqueous suspension or solution. In the preparation of tablets intended for oral administration, the carriers commonly used are lactose and corn starch. In addition, slip lubricants, such as magnesium stearate, are usually added. Convenient diluents used in the preparation of capsules for oral administration include lactose and dry corn starch. In case aqueous suspensions are used for oral administration, the active ingredient is combined with emulsifying and suspending agents. Certain sweetening, flavoring and / or coloring agents can be added if desired.

Compounds provided by the present invention can also be used in the form of suppositories for rectal administration of an appropriate drug. Similar formulations can be prepared by mixing said drug with an acceptable non-irritating excipient, which is characterized by a solid state at room temperature but a liquid state at the temperature of the rectum, as a result of which the excipient in question should melt in the rectum and release the drug. These substances include cocoa butter, beeswax, and polyethylene glycols.

Compounds of the present invention may also be administered topically, especially if the symptoms requiring treatment affect areas or organs that are completely accessible for topical treatment; such diseases include neurological diseases of the eyes, skin, or lower intestinal tract. Appropriate formulations intended for topical application can be prepared for each of these parts of the body.

When administered through the mucous membrane of the eye, these compounds can be used in the form of microscopically thin suspensions in isotonic sterile saline adjusted to pH or, in the preferred case, as solutions in isotonic sterile saline adjusted to pH, possibly containing preservatives, such as benzylalkonium chloride. In an alternative case of administration through the mucous membrane of the eye, the considered compounds can be used as part of an ointment, for example, based on petroleum jelly.

When applied topically to the skin, these compounds can be used in the form of an acceptable ointment, including the corresponding compound, suspended or dissolved, for example, in a mixture containing one or more of the following substances: mineral oil, liquid petrolatum, white petrolatum, propylene glycol, polyoxyethylene or polyoxypropylene a compound emulsifying wax, as well as water. Alternatively, the subject compound can be used in the form of an acceptable lotion or cream, including the corresponding active compound, suspended or dissolved in a mixture containing, for example, one or more of the following substances: mineral oil, sorbitan monostearate, polysorbate 60, cetyl ether wax , cetearyl alcohol, 2-octyldodecanol, benzyl alcohol, as well as water.

Local administration through the lower intestinal tract can be achieved using a preparation that is a rectal suppository (see above), or using an acceptable composition administered through an enema.

For the treatment of the aforementioned symptoms, these compounds, which are the active ingredient, can be used in doses ranging from 0.1 mg to 10,000 mg; preferred doses range from 0.1 mg to 1,000 mg. The specific amount of active ingredient mixed with the appropriate carrier materials upon receipt of a single dosage form depends on the characteristics of the particular patient, as well as the route of administration.

However, it is well known that the specific dose level for each individual patient depends on a number of factors, including the activity of the specific compound used, age, body weight, general health, gender, type of diet, duration of use, excretion rate, combination of active ingredients, the severity of the specific disease, the treatment of which is carried out by the specified compound, as well as the route of administration.

The subject compounds may be administered in conjunction with other neurotrophic agents, such as neurotrophic growth factor (NGF), glial growth factor, cerebral growth factor, ciliary neurotrophic factor, as well as neurotropin-3. The dose amount of such active ingredients having neurotrophin activity should depend on the above parameters, as well as on the effectiveness of the combination of active ingredients used.

The method for determining K _i

The ability of the compounds of the present invention to inhibit said peptidyl-prolyl-isomerase (rotamase) activity can be evaluated using well-known methods described in the literature (Harding, MW et al. Nature 341: 758-760 (1989); Holt et al. J. Am. Chem. Soc. 115: 9923-9938). The corresponding values, expressed as inhibition constants (K _i ), are presented in table 1. The intensity of cis-trans isomerization of the bond formed between the alanine and proline residues is spectroscopically evaluated on a model substrate, N-succinyl-Ala-Ala-Pro-Fen- p-nitroanilide using a test with bound chymotrypsin to release para-nitroanilide from the trans form of the substrate. The inhibition rate of this reaction due to the addition of various concentrations of the inhibitor is measured, and then the data are analyzed, considering the change in the first order of the rate constant as a function of the concentration of the inhibitor, with obtaining rounded values of K _i .

950 ml of ice test buffer (25 mM HEPES, pH 7.8; 100 mM NaCl), 10 ml FKBP solution (2.5 mM in 10 mM Tris-HCl, pH 7.5; 100 mM NaCl; 1 mM dithiothreitol), 25 ml of a chymotrypsin preparation (50 mg / ml in 1 mM HCl), as well as the test compound dissolved in various concentrations in 10 ml of dimethyl sulfoxide. Initiate the reaction by adding 5 ml of substrate (succinyl-Ala-Phen-Pro-para-nitroanilide, 5 mg / ml in 2.35 mm LiCl in trifluoroethanol).

Using a spectrophotometer, the absorption intensity at 390 nm is recorded for 90 s, and the reaction rate constants are determined based on the data characterizing the temporal dynamics of the absorption intensity.

The results obtained in these experiments are presented in table 1.

Table 1

No. R R ' K _i one 1,1-dimethylpropyl 3-phenylpropyl 42 2 - "- 3-phenyl-prop-2- (E) -enyl 125 3 - "- 3- (3,4,5-trimethoxyphenyl) propyl 200 four - "- 3- (3,4,5-trimethoxyphenyl) prop - 2- (E) -enyl 65 5 - "- 3- (4,5-methylenedioxy) phenylpropyl 170 6 - "- 3- (4,5-methylenedioxy) phenylprop-2- (E) -enyl 160 7 - "- 3-cyclohexylpropyl 200 8 - "- 3-cyclohexylprop-2- (E) -enyl 600 9 - "- (1R) -1,3-diphenyl-1-propyl 52 10 2-furanyl 3-phenylpropyl 4000 eleven 2-thienyl - "- 92 12 2-thiazolyl - "- one hundred 13 phenyl - "- 1970 fourteen 1,1-dimethylpropyl 3- (2,5-dimethoxy) phenylpropyl 250 fifteen - "- 3- (2,5-dimethoxy) phenylprop-2 (E) -enyl 450 16 - "- 3- (3,4,5-trimethoxyphenyl) ethyl 120 17 - "- 3- (3-pyridyl) propyl 5 eighteen - "- 3- (2-pyridyl) propyl 195 19 - "- 3- (4-pyridyl) propyl 23 twenty cyclohexyl 3-phenylpropyl 82 21 tert-butyl - "- 95 22 cyclohexylethyl - "- 1025 23 cyclohexylethyl 3- (3-pyridyl) propyl 1400 24 tert-butyl 3- (3-pyridyl) propyl 3 25 1,1-dimethylpropyl 3,3-diphenylpropyl 5 26 cyclohexyl 3- (3-pyridyl) propyl 9 27 2-thienyl 3- (3-pyridyl) propyl 1000 28 tert-butyl 3,3-diphenylpropyl 5 29th cyclohexyl - "- twenty thirty 2-thienyl - "- 150

In mammalian cells, FKBP-12 forms complexes with an inositol triphosphate receptor (IP ₃ R), as well as a ryanodine receptor (RyR). It is believed that the neurotrophic compounds provided by the present invention contribute to the dissociation of FKBP-12 from these complexes, as a result of which the corresponding calcium channel begins to “leak” (Cameron et al., 1995). Calcium fluxes are involved in the regulation of axon elongation, thus, inositol triphosphate and ryanodine receptors can participate in the implementation of the neurotrophic effects characteristic of the drugs in question. Since these drugs bind to the same site of the FKBP-12 molecule as the IP ₃ R receptor, it can be assumed that the drugs in question displace channels from FKBP-12.

Chicken spinal node cultures and axon extension

Isolation of the spinal nodes from the embryos of the chicken was carried out on the tenth day after fertilization. Entire ganglia explants were cultured at 37 ° C in an atmosphere with a 5% CO ₂ content; cultivation was carried out in 12-nested tablets coated with a thin Matrigel layer on Liebovitz L15 medium plus glucose supplemented with 2 mM glutamine and 10% fetal calf serum, as well as 10 μM cytosine-β-D arabinofuranoside (Ara C ) After 24 hours, said DRG's were treated with various concentrations of nerve growth factor, immunophilin ligands, or NGF + investigational drug combinations. 48 hours after the above drug treatment, a visual examination of the test ganglia was performed under an inverted Zeiss Axiovert microscope using phase contrast or Hoffman Modulation contrast. Microphotographs of the explants studied were obtained and axon elongation was measured. Axons whose length exceeded the DRG diameter were considered as positive; in addition, the total number of axons in each of the experimental variants was taken into account. From 3 to 4 DRG's were cultured in one well, each treatment option was performed in duplicate.

The data obtained in the above experiments are presented in table 2. Representative micrographs obtained using the compounds of example 17 are presented in figure 1; the dose-response curve for this compound is shown in FIG.

table 2
Axon extension in chicken DRG Example No. ED ₅₀ , axon elongation, nm one 53 2 105 3 149 four 190 5 10 6 75 10 0.46 eleven 0.015 fourteen 2 fifteen 0.8 16 0.015 17 0.05 eighteen thirty 19 6 twenty 0.13 21 0,025 22 0.66 23 1100 24 0.014 25 0.50 26 2 27 500 28 0.50 29th 10 thirty one hundred

Sciatic nerve axotomy

In anesthetized six-week-old male Sprague-Dawley rats, the sciatic nerve was exposed at the hip level and squeezed using tweezers. Immediately prior to the application of this damage to the rats, the test compounds or vehicle were subcutaneously administered, after which this administration was repeated daily for an additional 18 days. Slices of the damaged sciatic nerve were obtained, stained with silver dye Holmes (to determine the number of axons), and Luxol fast blue (to quantify the intensity of myelination). In animals treated with the vehicle, a significant decrease in the number of axons (a 50% decrease in relation to the intact control), as well as the intensity of myelination (90% decrease in relation to the intact control) was observed 18 days after the initial damage.

Compared with carrier treated animals, in experimental rats, which immediately before the damage was applied, and also daily for 18 days after the damage was done, subcutaneous administration of the compound from Example 1 (30 mg / kg) was carried out, reliable regeneration as the number of axons was observed ( 5% decrease in relation to the intact control), as well as myelination intensity (50% decrease in relation to the control). The significant efficacy of the compound of Example 1 is in good agreement with its potent ability to inhibit rotamase activity and stimulate axon extension in chicken DRG. These results are presented in figure 3. The designation "imitation" corresponds to control animals that received the carrier in the absence of damage. The designation "carrier" corresponds to animals that were injured and only the carrier substance was administered without the drug. Animals treated with the compound of Example 1 turned out to be very similar in the studied parameters to control animals that underwent imitation of damage, which indicates the powerful neuroregenerative effects exerted by this compound in vivo. As an inactive compound, a substance that did not exhibit inhibitory activity against FKBP-12 was used. Animals treated with an inactive compound resembled damaged animals that were administered with a carrier substance; the data obtained are in good agreement with the notion that the neuroregenerative properties of the compound of Example 1 are directly related to its inhibitory effect on FKBP-12. The obtained numerical values are presented in table 3.

Table 3 Type of processing The number of axons (% of control) Myelination (% of control) Imitation one hundred one hundred Damage + carrier (subcutaneous) fifty 10 + connection from Example 1
(30 mg / kg s.c.) one hundred fifty + inactive. conn. (30 mg / kg s.c.) 25 25

Modeling Parkinson's disease in mice using MRTR

To model Parkinson's disease in mice, damage to dopaminergic neurons using MRTP was used. Four-week-old white mice of the CD1 line were injected intraperitoneally with MPTR at a dose of 30 mg / kg for 5 days. During these 5 days, the experimental mice were further subcutaneously injected with either the compound of Example 17 (at a concentration of 10-40 mg / kg) or vehicle, after which the administration of the corresponding compound was continued for another 5 days. Eighteen days after the MPTP treatment, the experimental animals were sacrificed, the striatum was isolated, and it was homogenized. In order to evaluate the effect of the initial damage and drug treatment on the number of dopamine transporters (DAT), the binding of [ ³ H] CFT (radioactive ligand for the dopamine transporter) to striatal membranes was investigated. In order to quantify the survival and recovery of dopaminergic neurons, immunostaining using anti-tyrosine hydroxylase (TH) lg was performed on sagittal and coronal sections of the brain. In animals co-treated with MPTP and vehicle, a significant decrease in the number of functional dopaminergic endings was observed compared to intact animals. In damaged animals treated with the compound of Example 17, an almost complete restoration of the number of dopaminergic neurons stained with TH was observed.

Figures 4 and 5 show quantitative data on the content of DAT; 6-8 are micrographs illustrating the regenerative effects exerted in this model by the compound of Example 17. As follows from FIG. 4, the compounds provided by the present invention cause a substantial restoration of functional dopaminergic endings detected by [ ³ H] CFT binding, compared to MPTP treatment in the absence of Guilford compounds. In Fig. 5, the same data is presented in the form of a histogram. It was shown that for animals treated with MPTP and the compound of Example 17 (40 mg / kg), more than 90% recovery of the control level of [ ³ H] CFT binding was characteristic. As follows from Fig.6-8, immunostaining for tyrosine hydroxylase (a marker of viable dopaminergic neurons) in the striatum, black body, and also in the bundle of the middle part of the forebrain indicates a clear and significant restoration of functional neurons in animals treated with the compound of example 17, compared with animals treated with a damaging agent in the absence of an active compound (MPTP + carrier).

The following examples are intended to illustrate preferred embodiments of the present invention and should not be construed as limiting the present invention. All preferred embodiments of the present invention should not be construed as limiting the present invention. By the molecular weight of all polymers is meant the average molecular weight. In all cases, except where otherwise indicated, percentages are calculated as weight percentages relative to the final weight of the introduced system or the prepared composition; the total weight in all cases is 100%.

EXAMPLES

Compounds envisioned by the present invention can be prepared using various synthetic options using well-known chemical transformations. The main method for preparing the compounds provided by the present invention is shown in Scheme 1. Derivatives of N-glyoxyl-proline can be prepared by reacting methyl ester of L-proline with methyloxalyl chloride, as shown in Scheme 1. The resulting oxamates can be converted into interaction with various carbon nucleophiles to obtain intermediate compounds. Then, these intermediates are reacted with various alcohols, amides, or protected amino acid residues to give propyl esters or amides of the present invention.

DIAGRAM 1

EXAMPLE 1

Synthesis of 3-phenyl-1-propyl (2S) -1- (3,3-dimethyl-1,2-dioxo-pentyl) -2-pyrrolidinecarboxylate (Example 1).

but. Synthesis of Methyl (2S) -1- (1,2-dioxo-2-methoxyethyl) -2-pyrrolidine carboxylate

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 equivalents). The resulting suspension was stirred under a nitrogen atmosphere for 15 minutes, after which a solution of methyloxalyl chloride (3.20 g; 26.12 mmol) in methylene chloride (45 ml) was added dropwise. The resulting mixture was stirred at 0 ° C. for 1.5 hours. By removing the solid by filtration, the organic phase was washed with water, dried over MgSO ₄ and concentrated. The resulting crude residue was purified on a silica gel column, eluting with 50% ethyl acetate in hexane to yield 3.52 g (88%) of the product as a reddish oil. A mixture of cis-trans amide rotamers; data for trans rotamer are given

¹ H NMR (CDCl ₃ ): 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).

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

A solution of methyl (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 treated 14.2 ml of a solution of 1,1-dimethylpropyl magnesium chloride (1.0 M) in THF. The resulting homogeneous mixture was stirred at -78 ° C for 3 hours, after which the mixture was poured into a saturated solution of ammonium chloride (100 ml) and extracted with ethyl acetate. The resulting organic phase was washed with water, dried and concentrated, and the crude residue resulting from the removal of the aforementioned solvent was purified on a silica gel column, eluting with 25% ethyl acetate in hexane to yield 2.10 g (75%) of oxamate as a colorless oil.

¹ H NMR (CDCl ₃ ): d 0.88 (t, 2H); 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).

at. Synthesis of (2S) -1- (1,2-dioxo-3,3-dimethylphenylpentyl) -2-pyrrolidinecarboxylic acid

A mixture of methyl (2S) -1- (1,2-dioxo-3,3-dimethylphenylpentyl) -2-pyrrolidinecarboxylate (2.10 g; 8.23 mmol), 1 N LiOH (15 ml) and methanol (50 ml) stirred at a temperature of 0 ° C for 30 minutes, and then overnight at room temperature. The specified mixture using 1 N. HCl was acidified to pH 1, diluted with water and extracted with 100 ml of methylene chloride. The resulting organic extract was washed with brine and concentrated to yield 1.73 g (87%) of a snow-white solid that did not require further purification.

¹ H NMR (CDCl ₃ ): d 0.87 (t, 3H); 1.22; 1.25 (s, 3H each); 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).

The synthesis of 3-phenyl-1-propyl (2S) -1- (3,3-dimethyl-1,2-dioxo-pentyl) -2-pyrrolidinecarboxylate (example 1)

A mixture of (2S) -1- (1,2-dioxo-3,3-dimethylphenylpentyl) -2-pyrrolidine-carboxylic acid (600 mg; 2.49 mmol), 3-phenyl-1-propanol (508 mg; 3, 73 mmol) of dicyclohexylcarbodiimide (822 mg; 3.98 mmol), camphorsulfonic acid (190 mg; 0.8 mmol) and 4-dimethylaminopyridine (100 mg; 0.8 mmol) in methylene chloride (20 ml) were stirred overnight in the atmosphere nitrogen. A solid precipitate was removed from the resulting reaction mixture by filtration through brownmillerite, the obtained filtrate was concentrated in vacuo, and the isolated crude material was purified by flash column chromatography (elution with 25% ethyl acetate in hexane) in 720 mg (80%) of the above. compounds in the form of a colorless oil.

¹ H NMR (CDCl ₃ ): d 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 above in example 1 was used to obtain the following compounds, illustrating the present invention:

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

80% ¹ H NMR (360 MHz, CDCl ₃ ): d 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 (d, 1H); 6.67 (dd, 1H, J = 15.9) 7.13-7.50 (m, 5H).

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

61%; ¹ H NMR (CDCl ₃ ): d 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 4: 3- (3,4,5-trimethoxyphenyl) -1-prop-2- (E) -enyl (2S) -1- (3,3-dimethyl-1,2-dioxopentyl) -2-pyrrolidin- carboxylate

66% ¹ H NMR (CDCl ₃ ): d 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 (d, 1H); 6.58 (d, 1H, J = 16); 6.63 (s, 2H).

Example 5: 3- (4,5-methylenedioxyphenyl) -1-propyl (2S) -1- (3,3-dimethyl-1,2-dioxopentyl) -2-pyrrolidinecarboxylate

82%; ¹ H NMR (360 MHz, CDCl ₃ ): d 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 (d, 1H); 6.57 (dd, 1H, J = 1.3; 8.0); 6.93 (s, 1H).

Example 6: 3- (4,5-methylenedioxyphenyl) -1-prop-2- (E) -enyl (2S) -1-3,3-dimethyl-1,2-dioxopentyl) -2-pyrrolidinecarboxylate

82%; ¹ H NMR (360 MHz, CDCl ₃ ): d 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); 4.53 (dd, 1H, J = 3.8; 8.6) 4.61-4.89 (m, 2H); 5.96 (s, 2H); 6.10 (d, 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 8: 3-cyclohexyl-1-prop-2- (E) -enyl (2S) -1- (3,3-dimethyl-1,2-dioxopentyl) -2-pyrrolidinecarboxylate

92%; ¹ H NMR (360 MHz, CDCl ₃ ): 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, J = 6.5; 16).

Example 9: (1R) -1,3-diphenyl-1-propyl (2S) -1- (3,3-dimethyl-1,2-dioxopentyl) -2-pyrrolidinecarboxylate

90% ¹ H NMR (360 MHz, CDCl ₃ ): d 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 10: 3-phenyl-1-propyl (2S) -1- (1,2-dioxo-2- [2-furanyl]) - ethyl-2-pyrrolidinecarboxylate

99%; ¹ H NMR (300 MHz, CDCl ₃ ): d 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 11: 3-phenyl-1-propyl (2S) -1- (1,2-dioxo-2- [2-thienyl]) - ethyl-2-pyrrolidinecarboxylate

81% ¹ H NMR (300 MHz, CDCl ₃ ): d 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 13: 3-phenyl-1-propyl (2S) -1- (1,2-dioxo-2-phenyl) ethyl-2-pyrrolidinecarboxylate

99%; ¹ H NMR (300 MHz, CDCl ₃ ): d 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 14: 3- (2,5-dimethoxyphenyl) -1-propyl (2S) -1- (3,3-dimethyl-1,2-dioxopentyl) -2-pyrrolidinecarboxylate

99%; ¹ H NMR (300 MHz, CDCl ₃ ): d 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 15: 3- (2,5-dimethoxyphenyl) -1-prop-2- (E) -enyl (2S) -1-3,3-dimethyl-1,2-dioxopentyl) -2-pyrrolidine-carboxylate

99%; ¹ H NMR (300 MHz, CDCl ₃ ): d 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 16: 3- (3,4,5-trimethoxyphenyl) -1-ethyl (2S) -1- (3,3-dimethyl-1,2-dioxopentyl) -2-pyrrolidinecarboxylate

97%; ¹ H NMR (300 MHz, CDCl ₃ ): d 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 (m, 2H); 3.79 (s, 3H); 3.83 (s, 3H); 4.14 (m, 2H); 4.52 (d, 1H); 6.36 (m, 1H).

Example 17: 3- (3-pyridyl) -1-propyl (2S) -1- (3,3-dimethyl-1,2-dioxopentyl) -2-pyrrolidinecarboxylate

80% ¹ H NMR (300 MHz, CDCl ₃ ): d 0.85 (t, 3H); 1.23; 1.26 (s, 3H each); 1.63-1.89 (m, 2H); 1.90-2.30 (m, 4H); 2.30-2.50 (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 18: 3- (2-pyridyl) -1-propyl (2S) -1- (3,3-dimethyl-1,2-dioxopentyl) -2-pyrrolidinecarboxylate

88% ¹ H NMR (300 MHz, CDCl ₃ ): d 0.84 (t, 3H); 1.22; 1.27 (s, 3H each); 1.68-2.32 (m, 2H); 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 19: 3- (4-pyridyl) -1-propyl (2S) -1- (3,3-dimethyl-1,2-dioxopentyl) -2-pyrrolidinecarboxylate

91% ¹ H NMR (300 MHz, CDCl ₃ ): d 0.89 (t, 3H; J = 7.5); 1.23 (s, 3H); 1.25 (s, 3H); 1.22-1.85 (m, 7H); 2.33 (m, 1H); 3.30 (m, 2H); 3.79 (s, 3H); 4.08 (s, 3H); 4.12 (m, 1H); 5.25 (d, 1H; J = 5.7); 6.28 (m, 1H); 6.92-6.80 (m, 4H).

Example 20: 3-phenyl-1-propyl (2S) -1- (2-cyclohexyl-1,2-dioxoethyl) -2-pyrrolidinecarboxylate

91% ¹ H NMR (300 MHz, CDCl ₃ ): d 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 21: 3-phenyl-1-propyl (2S) -1- (2-tert-butyl-1,2-dioxoethyl) -2-pyrrolidinecarboxylate

92%; ¹ H NMR (300 MHz, CDCl ₃ ): d 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 22: 3-phenyl-1-propyl (2S) -1- (2-cyclohexylethyl-1,2-dioxoethyl) -2-pyrrolidinecarboxylate

97%; ¹ H NMR (300 MHz, CDCl ₃ ): d 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 (m, 1H); 7.17-7.32 (m, 5H).

Example 23: 3- (3-pyridyl) -1-propyl (2S) -1- (2-cyclohexylethyl-1,2-dioxopentyl) -2-pyrrolidinecarboxylate

70% ¹ H NMR (300 MHz, CDCl ₃ ): d 0.87 (m, 2H); 1.16 (m, 4H); 1.49 (m, 2H); 1.68 (m, 4H); 1.95-2.32 (m, 7H); 2.71 (m, 2H); 2.85 (m, 2H); 3.63-3.78 (m, 2H); 4.19 (m, 2H); 5.30 (m, 1H); 7.23 (m, 1H); 7.53 (m, 1H); 8.46 (m, 2H).

Example 24: 3- (3-pyridyl) -1-propyl (2S) -1- (2-tert-butyl-1,2-dioxoethyl) -2-pyrrolidinecarboxylate

83%; ¹ H NMR (300 MHz, CDCl ₃ ): d 1.29 (s, 9H); 1.95-2.04 (m, 5H); 2.31 (m, 1H); 2.72 (t, 2H; J = 7.5); 3.52 (m, 2H); 4.18 (m, 2H); 4.52 (m, 1H); 7.19-7.25 (m, 1H); 7.53 (m, 1H); 8.46 (m, 2H).

Example 25: 3,3-diphenyl-1-propyl (2S) -1- (3,3-dimethyl-1,2-dioxopentyl) -2-pyrrolidinecarboxylate

99%; ¹ H NMR (300 MHz, CDCl ₃ ): d 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 26: 3- (3-pyridyl) -1-propyl (2S) -1- (2-cyclohexyl-1,2-dioxopentyl) -2-pyrrolidinecarboxylate

88% ¹ H NMR (300 MHz, CDCl ₃ ): d 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 27: 3- (3-pyridyl) -1-propyl (2S) -N - ([2-thienyl] glyoxyl) pyrrolidinecarboxylate

49% ¹ H NMR (300 MHz, CDCl ₃ ): d 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, 1H); 7.61 (m, 1H); 7.80 (d, 1H); 8.04 (d, 1H); 8.46 (m, 2H).

Example 28: 3,3-diphenyl-1-propyl (2S) -1- (3,3-dimethyl-1,2-dioxobutyl) -2-pyrrolidinecarboxylate

99%; ¹ H NMR (300 MHz, CDCl ₃ ): d 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 29: 3,3-diphenyl-1-propyl (2S) -1-cyclohexylglyoxyl-2-pyrrolidinecarboxylate

91% ¹ H NMR (300 MHz, CDCl ₃ ): d 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 30: 3,3-diphenyl-1-propyl (2S) -1- (2-thienyl) glyoxyl-2-pyrrolidinecarboxylate

75% ¹ H NMR (300 MHz, CDCl ₃ ): 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, 1H); 7.25 (m, 11H); 7.76 (dd, 1H); 8.03 (m, 1H).

By the method described above in example 1, the following compounds were also obtained.

Example 31: (2S) -1- (3,3-dimethyl-1,2-dioxopentyl) -2-pyrrolidinecarboxylic acid

¹ H NMR (CDCl ₃ , 400 MHz): δ 0.86 (t, 3H, J = 8.0); 1.21 (s, 3H); 1.23 (s, 3H); 1.74 (m, 2H); 2.23 (m, 2H); 2.31 (m, 2H); 3.49 (m, 2H); 4.59 (dd, 1H, J = 8.4, 3.8). Elemental analysis: C: 59.73 (59.60), H: 7.94 (7.95), N: 5.80 (5.66).

Example 32: (2S) -1- (2-cyclohexyl-1,2-dioxoethyl) -2-pyrrolidinecarboxylic acid

¹ H NMR (CDCl ₃ , 400 MHz): δ 1.40-1.45 (m, 6H); 1.57-1.60 (m, 2H); 1.82 (m, 2H); 1.70-1.74 (m, 4H); 2.30 (dm, 1H); 3.35 (m, 2H); 4.22 (d, 1H); 11.00 (br, 1H).

Example 33: (2S) -1- (2-cyclohexylethyl-1,2-dioxoethyl) -2-pyrrolidinecarboxylic acid

¹ H NMR (CDCl ₃ , 400 MHz): δ 1.38-1.40 (m, 4H); 1.44-1.48 (m, 6H); 1.55-1.57 (m, 2H); 1.59 (m, 2H); 1.80-1.83 (m, 4H); 2.42 (dm, 1H); 3.31 (m, 2H); 4.25 (d, 1H); 11.20 (br, 1H).

Example 34: (2S) -1- (1,2-dioxo-2- [2-furanyl]) ethyl-2-pyrrolidinecarboxylic acid

¹ H NMR (CDCl ₃ , 400 MHz): δ 1.60 (m, 2H); 1.80-1.81 (m, 2H); 3.35 (m, 2H); 4.23 (d, 1H); 6.61 (d, 1H); 7.23 (d, 1H); 7.72 (d, 1H); 10.90 (br, 1H).

Example 35: (2S) -1- (1,2-dioxo-2- [2-thienyl]) ethyl-2-pyrrolidinecarboxylic acid

¹ H NMR (CDCl ₃ , 400 MHz): δ 1.60 (m, 2H); 1.83 (m, 2H); 3.32 (m, 2H); 4.21 (d, 1H); 7.06 (d, 1H); 7.61 (d, 1H); 7.65 (d, 1H); 11.20 (br, 1H).

Example 36: (2S) -1- (1,2-dioxo-2- [2-thiazolyl]) ethyl-2-pyrrolidinecarboxylic acid

¹ H NMR (CDCl ₃ , 400 MHz): δ 1.59 (m, 2H); 1.82 (m, 2H); 3.35 (m, 2H); 4.22 (d, 1H); 7.86 (d, 1H); 8.08 (d, 1H); 11.00 (br, 1H).

Example 37: (2S) -1- (1,2-dioxo-2-phenyl) ethyl-2-pyrrolidinecarboxylic acid

¹ H NMR (CDCl ₃ , 400 MHz): δ 1.61 (m, 2H); 1.85 (m, 2H); 3.36 (m, 2H); 4.24 (d, 1H); 7.81 (m, 2H); 7.39-7.45 (m, 3H); 11.08 (br, 1H).

Example 38: (2S) -1- (3,3-dimethyl-1,2-dioxopentyl) -2-pyrrolidinecarboxamide

¹ H NMR (CDCl ₃ , 400 MHz): δ 0.90 (t, 3H, J = 7.5); 1.22 (s, 6H); 1.74 (m, 2H); 1.85-2.15 (m, 3H); 2.38 (m, 1H); 3.47 (m, 2H); 4.60 (m, 1H); 5.60 (bs, 1H); 6.75 (bs, 1H).

Analysis: Calculated for C ₁₂ H ₂₀ N ₂ O ₃ : C, 59.98; H, 8.39; N, 11.66.

Found: C, 59.75; H, 8.37; N, 11.40.

TLC: R _f = 0.38 (100% EtOAc).

Example 39: (2S) -1- (2-tert-butyl-1,2-dioxoethyl) -2-pyrrolidinecarboxylic acid

¹ H NMR (CDCl ₃ , 400 MHz): δ 1.28 (s, 9H); 1.95-2.06 (m, 2H); 2.17-2.26 (m, 2H); 3.48-3.52 (m, 2H); 4.52 (d, 1H); 7.95 (br, 1H).

Analysis: Calculated for C ₁₁ H ₁₇ NO ₄ : C, 58.14; H, 7.54; N, 6.16. Found: C, 58.40; H, 7.56; N, 6.14.

The necessary substituted alcohols can be obtained using a number of methods well known to specialists in the field of organic synthesis. As shown in Scheme 2, alkyl or aryl aldehydes can be homogenized to phenylpropanols by reaction with methyl (triphenylphosphoranilidene) acetate to give various trans-cinnamates; then, the indicated cinnamates can be reduced to saturated alcohols, either by reacting them with an excess of lithium hydride and aluminum, or by performing their phased reduction: first, reduction of the double bond (by catalytic hydrogenation), and then reduction of the obtained saturated ether (using appropriate reducing agents). Alternatively, the trans-cinnamates in question can be reduced to (E) -allyl alcohols using diisobutylaluminum hydride.

Alcohols with a long hydrocarbon chain can be obtained by homogenization of benzyl and higher aldehydes. Alternatively, these aldehydes can be obtained from the corresponding phenylacetic and higher acids, as well as from phenylethyl and higher alcohols.

The main method for the synthesis of acrylic esters, in particular methyl (3,3,5-trimethoxy) -trans-cinnamate, is as follows:

A solution of 3,4,5-trimethoxybenzaldehyde (5.0 g; 25.48 mmol) and methyl (triphenylphosphoranilidene) acetate (10.0 g; 29.91 mmol) in tetrahydrofuran (250 ml) was refluxed overnight. Then the indicated reaction mixture was cooled, diluted with 200 ml of ethyl acetate, washed with water (2 × 200 ml), dried and concentrated in vacuo. The resulting crude residue was purified by silica gel column chromatography, eluting with 25% ethyl acetate in hexane, to give 5.63 g (88%) of the title cinnamate as a crystalline solid.

¹ H NMR (300 MHz, CDCl ₃ ): d 3.78 (s, 3H); 3.85 (s, 6H); 6.32 (d, 1H; J = 16); 6.72 (s, 2H); 7.59 (d, 1H; J = 16).

The main method for the synthesis of saturated alcohols using the example of (3,4,5-trimethoxy) phenylpropanol is as follows:

To a solution of lithium and aluminum hydride (14 mM) in THF (35 ml), stirred under argon, was added dropwise a solution of methyl (3,3,5-trimethoxy) trans cinnamate (1.81 g; 7.17 mmol) ) in tetrahydrofuran (30 ml). Upon completion of the indicated addition, the resulting mixture was kept at a temperature of 75 ° С for 4 h, cooled, and then the reaction was stopped by carefully adding 15 ml of 2 N NaOH and then 50 ml of water. The resulting mixture was filtered through brownmillerite to remove a solid precipitate, and the used filter was washed with ethyl acetate. The obtained organic fractions were combined, washed with water, dried, concentrated in vacuo and purified by chromatography on a silica gel column, eluting with ethyl acetate in 0.86 g (53%) of the above-mentioned alcohol as a clear oil.

¹ H NMR (300 MHz, CDCl ₃ ): d 1.23 (br, 1H); 1.87 (m, 2H); 2.61 (t, 2H; J = 7.1); 3.66 (t, 2H); 3.80 (s, 3H); 3.83 (s, 6H); 6.40 (s, 2H).

The main method for the synthesis of trans-allyl alcohols using the example of (3,4,5-trimethoxy) phenylprop-2- (E) -enol from acrylic esters is as follows:

A solution of (3,3,5-trimethoxy) -trans-cinnamate (1.35 g; 5.35 mmol) in toluene (25 ml) was cooled to a temperature of -10 ° C, after which it was treated with a solution of diisobutylaluminum hydride (11.25 mmol) ) in toluene (11.25 ml of a 1.0 M solution, 11.25 mmol). The resulting reaction mixture was stirred at 0 ° C for 3 hours, after which the reaction was stopped by adding 3 ml of methanol, and then 1 N. HCl to pH 1. The reaction mixture was extracted with ethyl acetate, the resulting organic fraction was washed with water, dried and concentrated. Purification by chromatography on a silica gel column (elution with 25% ethyl acetate in hexane) gave 0.96 g (80%) of the title compound as a thick oil.

¹ H NMR (360 MHz, CDCl ₃ ): d 3.85 (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).

It is obvious that the synthesis described in the present invention can be carried out with a wide variety of variations. All such variations should not be perceived in isolation from the present invention, but, on the contrary, should be considered as part of the following claims.

Claims (8)

1. The compound of the formula:

Where R ₁ is C ₁ -C ₉ straight or branched chain alkyl optionally substituted with C ₃ -C ₈ cycloalkyl, C ₆ cycloalkyl, 2-furyl, 3-furyl, 2-thiazolyl, 2-thienyl, 3-thienyl or phenyl; X is oxygen; Y is oxygen or NR ₂ , where R ₂ is hydrogen or C ₁ -C ₆ alkyl; and Z is hydrogen; provided that, when X is oxygen and Y is oxygen, then R _{1 is} not methyl, ethyl, isopropyl, isobutyl or phenyl; when X is oxygen and Y is NR ₂ , where R ₂ is hydrogen, methyl, isopropyl or tert-butyl, then R _{1 is} not methyl. 2. The compound according to claim 1, having the formula:

where Z is hydrogen. 3. The compound according to claim 2, where R _{1 is} selected from the group consisting of straight chain or branched C ₁ -C ₉ alkyl, 2-cyclohexyl, 4-cyclohexyl, 2-furanyl, 2-thienyl and 2-thiazolyl. 4. The compound according to claim 1, having the formula:

where Z is hydrogen. 5. The compound of the formula:

Where R ₁ is C ₁ -C ₅ straight or branched chain alkyl optionally substituted with C ₃ -C ₆ cycloalkyl; or a group selected from the group consisting of 2-furyl, 2-thienyl and phenyl; X is oxygen; Y is oxygen; Z is hydrogen; provided that R _{1 is} not methyl, ethyl, isopropyl, isobutyl or phenyl. 6. The compound according to claim 1, which is selected from the group consisting of (2S) -1- (3,3-dimethyl-1,2-dioxopentyl) -2-pyrrolidinecarboxylic acid, (2S) -1- (2-cyclohexyl-1,2-dioxoethyl) -2-pyrrolidinecarboxylic acid, (2S) -1- (2-cyclohexylethyl-1,2-dioxoethyl) -2-pyrrolidinecarboxylic acid, (2S) -1- (1,2-dioxo-2- [2-furanyl]) ethyl-2-pyrrolidinecarboxylic acid, (2S) -1- (1,2-dioxo-2- [2-thienyl]) ethyl-2-pyrrolidinecarboxylic acid, (2S) -1- (1,2-dioxo-2- [2-thiazolyl]) ethyl-2-pyrrolidinecarboxylic acid; and (2S) -1- (3,3-dimethyl-1,2-dioxopentyl) -2-pyrrolidinecarboxamide. 7. The compound according to claim 5, selected from the group consisting of (2S) -1- (3,3-dimethyl-1,2-dioxopentyl) -2-pyrrolidinecarboxylic acid, (2S) -1- (2-cyclohexylethyl-1,2-dioxoethyl) -2-pyrrolidinecarboxylic acid, (2S) -1- (2-tert-butyl-1,2-dioxoethyl) -2-pyrrolidinecarboxylic acid, (2S) -1- (2-cyclohexyl-1,2-dioxoethyl) -2-pyrrolidinecarboxylic acid; and (2S) -1- (1,2-dioxo-2- [2-thienyl]) ethyl-2-pyrrolidinecarboxylic acid. 8. A chemical compound consisting of (2S) -1- (1,2-dioxo-3,3-dimethylpentyl) -2-pyrrolidinecarboxylic acid.

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