U.S. patent application number 10/177666 was filed with the patent office on 2003-02-13 for heterocyclic esters and amides.
Invention is credited to Hamilton, Gregory S., Li, Jia-He.
Application Number | 20030032635 10/177666 |
Document ID | / |
Family ID | 24892040 |
Filed Date | 2003-02-13 |
United States Patent
Application |
20030032635 |
Kind Code |
A1 |
Li, Jia-He ; et al. |
February 13, 2003 |
Heterocyclic esters and amides
Abstract
This invention relates to neurotrophic low molecular weight,
small molecule heterocyclic ester and amides having an affinity for
FKBP-type immunophilins, and their use as inhibitors of the enzyme
activity associated with immunophilin proteins, particularly
peptidyl-prolyl isomerase, or rotamase, enzyme activity.
Inventors: |
Li, Jia-He; (Cockeysville,
MD) ; Hamilton, Gregory S.; (Catonsville,
MD) |
Correspondence
Address: |
GUILFORD PHARMACEUTICALS C/O
FOLEY & LARDNER
3000 K STREET, NW
WASHINGTON
DC
20007-5143
US
|
Family ID: |
24892040 |
Appl. No.: |
10/177666 |
Filed: |
June 24, 2002 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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10177666 |
Jun 24, 2002 |
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09733043 |
Dec 11, 2000 |
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09733043 |
Dec 11, 2000 |
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09027622 |
Feb 23, 1998 |
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6200972 |
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09027622 |
Feb 23, 1998 |
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08719947 |
Sep 25, 1996 |
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5801187 |
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Current U.S.
Class: |
514/211.01 ;
514/217.11; 514/218; 514/227.5; 514/237.5; 514/255.01; 514/365;
514/374; 514/399; 540/544; 540/575; 544/171; 544/386; 544/59;
548/200; 548/215; 548/334.1 |
Current CPC
Class: |
C07D 233/02 20130101;
A61P 25/00 20180101; C07D 279/12 20130101; A61P 25/28 20180101;
C07D 265/28 20130101; C07D 417/12 20130101; C07D 277/06 20130101;
A61P 25/16 20180101; C07D 241/04 20130101 |
Class at
Publication: |
514/211.01 ;
514/217.11; 514/218; 514/227.5; 514/237.5; 514/255.01; 514/365;
514/374; 514/399; 540/544; 540/575; 544/59; 544/171; 544/386;
548/200; 548/215; 548/334.1 |
International
Class: |
A61K 031/554; A61K
031/553; A61K 031/551; A61K 031/54; A61K 031/538; A61K 031/537;
A61K 031/496; A61K 031/426; A61K 031/42; A61K 031/4172 |
Claims
What is claimed is:
1. A compound of formula I: 7or a pharmaceutically acceptable salt
thereof, wherein: A and B, together with the nitrogen and carbon
atoms to which they are respectively attached, form a 5-7 membered
saturated or unsaturated heterocyclic ring containing, in addition
to the nitrogen atom, at least one additional O, S, SO, SO.sub.2,
NH or NR.sub.1 heteroatom in any chemically stable oxidation state;
X is O or S; Z is O, NH or NR.sub.1; W and Y are independently O,
S, CH.sub.2 or H.sub.2; R.sub.1 is C.sub.1-C.sub.6 straight or
branched chain alkyl or alkenyl, which is substituted in one or
more position(s) with (Ar.sub.1).sub.n, (Ar.sub.1).sub.n connected
by a C.sub.1-C.sub.6 straight or branched chain alkyl or alkenyl,
C.sub.3-C.sub.8 cycloalkyl, C.sub.3-C.sub.8 cycloalkyl connected by
a C.sub.1-C.sub.6 straight or branches chain alkyl or alkenyl,
Ar.sub.2, or a combination thereof; n is 1 or 2; R.sub.2 is either
C.sub.1-C.sub.9 straight or branched chain alkyl or alkenyl,
C.sub.3-C.sub.8 cycloalkenyl, C.sub.5-C.sub.7 cycloalkenyl, or
Ar.sub.1, wherein said alkyl, alkenyl, cycloalkyl or cycloalkenyl
is either unsubstituted or substituted in one or more position(s)
with C.sub.1-C.sub.4 straight or branched chain alkyl or alkenyl,
hydroxyl, or a combination thereof; and Ar.sub.1 and Ar.sub.2 are
independently a mono-, bi- or tricyclic, carbo- or heterocyclic
ring, wherein the ring is either unsubstituted or substituted in
one to three position(s) with halo, hydroxyl, nitro,
trifluoromethyl, C.sub.1-C.sub.6 straight or branched chain alkyl
or alkenyl, C.sub.1-C.sub.4 alkoxy, C.sub.1-C.sub.4 alkenyloxy,
phenoxy, benzyloxy, amino, or a combination thereof; wherein the
individual ring sizes are 5-6 members; and wherein the heterocyclic
ring contains 1-6 heteroatom(s) selected from the group consisting
of O, N, S, and a combination thereof.
2. The compound of claim 1, wherein the mono- or bicyclic, carbo-
or heterocyclic ring is selected from the group consisting of
naphthyl, indolyl, furyl, thiazolyl, thienyl, pyridyl, quinolinyl,
isoquinolinyl, fluorenyl and phenyl.
3. The compound of claim 1, wherein the at least one additional
heteroatom in the 5-7 membered saturated or unsaturated
heterocyclic ring is NH or NR.sub.1.
4. The compound of claim 1, wherein the compound has an affinity
for FKBP-type immunophilins.
5. The compound of claim 4, wherein the FKBP-type immunophilin is
FKBP12.
6. The compound of claim 1, wherein the compound inhibits rotamase
enzyme activity.
7. A pharmaceutical composition comprising a neurotrophically
effective amount of the compound of claim 1 and a pharmaceutically
acceptable carrier.
8. A method of effecting a neuronal activity in an animal,
comprising: administering to the animal a neurotrophically
effective amount of the compound of claim 1.
9. The method of claim 8, wherein the neuronal activity is selected
from the group consisting of stimulation or damaged neurons,
promotion of neuronal regeneration, prevention of neurodegeneration
and treatment of neurological disorder.
10. The method or claim 9, wherein the neurological disorder is
selected from the group consisting of peripheral neuropathy caused
by physical injury or disease state, physical damage to the brain,
physical damage to the spinal cord, stroke associated with brain
damage, and neurological disorder relating to
neurodegeneration.
11. The method of claim 10, wherein the neurological disorder
relating to neurodegeneration is selected from the group consisting
of Alzheimer's Disease, Parkinson's Disease, and amyotrophic
lateral sclerosis.
12. A compound of formula II: 8or a pharmaceutically acceptable
salt thereof, wherein: A, B and C are independently CH.sub.2,O, S,
SO, SO.sub.2, NH or NH.sub.1; R.sub.1 is C.sub.1-C.sub.5 straight
or branched chain a alkenyl, which is substituted in one or more
position(s) with (Ar.sub.1).sub.n, (Ar.sub.1).sub.n connected by a
C.sub.1-C.sub.6 straight or branched chain alkyl or alkenyl, or a
combination thereof; n is 1 or 2; R.sub.2 is either C.sub.1-C.sub.9
straight or branched chain alkyl or alkenyl, C.sub.3-C.sub.9
cycloalkyl, C.sub.5-C.sub.7 cycloalkenyl, or Ar.sub.1; and Ar.sub.1
is a mono-, bi- or tricyclic, carbo- or heterocyclic ring, wherein
the ring is either unsubstituted or substituted in one to three
position(s) with halo, hydroxyl, nitro, trifluoromethyl,
C.sub.1-C.sub.6 straight or branched chain alkyl or alkenyl,
C.sub.1-C.sub.4 alkoxy, C.sub.1-C.sub.4 alkenyloxy, phenoxy,
benzyloxy, amino, or a combination thereof; wherein the individual
ring sizes are 5-6 members; and wherein the heterocyclic ring
contains 1-6 heteroatom(s) selected from the group consisting of O,
N, S, and a combination thereof.
13. The compound of claim 12, wherein: A is CH.sub.2; B is CH.sub.2
or S; C is CH.sub.2 or NH; R.sub.1 is selected from the group
consisting of 3-phenylpropyl and 3-(3-pyridyl)propyl; and R.sub.1
is selected from the group consisting of 3,3-dimethylpentyl,
cyclohexyl, and tert-butyl.
14. The compound of claim 13, wherein: B is CH.sub.2; C is NH; and
R.sub.1is 3-phenylpropyl.
15. The compound of claim 13, wherein: B is S; and C is
CH.sub.2.
16. The compound of claim 12, wherein the compound is selected from
the group consisting of:
3-phenyl-1-propyl(2S)-1-(3,3-dimethyl-1,2-dioxopenty-
l)-2-(4-thiazolidine)carboxylate; and
3-(3-pyridyl)-1-propyl(2S)-1-(3,3-di- methyl-1,
2-dioxopentyl)-2-(4-thiazolidine) carboxylate.
17. The compound of claim 12, wherein the compound has an affinity
for FKBP-type immunophilins.
18. The compound of claim 17, wherein the FKBP-type immunophilin is
FKBP12.
19. The compound of claim 12, wherein the compound inhibits
rotamase enzyme activity.
20. A pharmaceutical composition comprising a neurotrophically
effective amount of the compound of claim 12 and a pharmaceutically
acceptable carrier.
21. A method of effecting a neuronal activity in an animal,
comprising: administering to the animal a neurotrophically
effective amount of the compound of claim 12.
22. The method of claim 21, wherein the neuronal activity is
selected from the group consisting of stimulation of damaged
neurons, promotion of neuronal regeneration, prevention of
neurodegeneration and treatment of neurological disorder.
23. The method of claim 22, wherein the neurological disorder is
selected from the group consisting of peripheral neuropathy caused
by physical injury or disease state, physical damage to the brain,
physical damage to the spinal cord, stroke associated with brain
damage, and neurological disorder relating to
neurodegeneration.
24. The method of claim 23, wherein the neurological disorder
relating to neurodegeneration is selected from the group consisting
of Alzheimer's Disease, Parkinson's Disease, and amyotrophic
lateral sclerosis.
25. A compound of formula III: 9or a pharmaceutically acceptable
salt thereof, wherein: A, B, C and D are independently CH.sub.2, O,
S, SO, SO.sub.2, NH or NR.sub.1; R.sub.1 is C.sub.1-C.sub.5
straight or branched chain alkyl or alkenyl, which is substituted
in one or more position(s) With (Ar.sub.1).sub.n, (Ar.sub.1).sub.n
connected by a C.sub.1-C.sub.6 straight or branched chain alkyl or
alkenyl, or a combination thereof; n is 1 or 2; R.sub.2 is either
C.sub.1-C.sub.9 straight or branched chain alkyl or alkenyl,
C.sub.3-C.sub.8 cycloalkyl, C.sub.5-C.sub.7 cycloalkenyl, or
Ar.sub.1; and Ar.sub.1 is a mono-, bi- or tricyclic, carbo- or
heterocyclic ring, wherein the ring is either unsubstituted or
substituted in one to three position(s) with halo, hydroxyl, nitro,
trifluoromethyl, C.sub.1-C.sub.6 straight or branched chain alkyl
or alkenyl, C.sub.1-C.sub.4 alkoxy, C.sub.1-C.sub.4 alkenyloxy,
phenoxy, benzyloxy, amino, or a combination thereof; wherein the
individual ring sizes are 5-6 members; and wherein the heterocyclic
ring contains 1-6 heteroatom(s) selected from the group consisting
of O, N, S, and a combination thereof.
26. The compound of claim 25, wherein: A is CH.sub.2; B is
CH.sub.2; C is S, O or NH; D is CH.sub.2; R.sub.1 is selected from
the group consisting of 3-phenylpropyl and (3,4,5-trimethoxy)
phenylpropyl; and R.sub.2 is selected from the group consisting of
3,3-dimethylpentyl, cyclohexyl, 3-3-dimethylpropyl, phenyl, and
3,4,5-trimethoxyphenyl.
27. The compound of claim 26, wherein: C is NH; and R.sub.2 is
3,3-dimethylpropyl or phenyl.
28. The compound of claim 25, wherein the compound has an affinity
for FKBP-type immunophilins.
29. The compound of claim 28, wherein the FKBP-type immunophilin is
FKBP12.
30. The compound of claim 25, wherein the compound inhibits
rotamase enzyme activity.
31. A pharmaceutical composition comprising a neurotrophically
effective amount of the compound of claim 25 and a pharmaceutically
acceptable carrier.
32. A method of effecting a neuronal activity in an animal,
comprising: administering to the animal a neurotrophically
effective amount of the compound of claim 25.
33. The method of claim 32, wherein the neuronal activity is
selected from the group consisting of stimulation of damaged
neurons, promotion of neuronal regeneration, prevention of
neurodegeneration and treatment of neurological disorder.
34. The method of claim 33, wherein the neurological disorder is
selected from the group consisting of peripheral neuropathy caused
by physical injury or disease state, physical damage to the brain,
physical damage to the spinal cord, stroke associated with brain
damage, and neurological disorder relating to
neurodegeneration.
35. The method of claim 34, wherein the neurological disorder
relating to neurodegeneration is selected from the group consisting
of Alzheimer's Disease, Parkinson's Disease, and amyotrophic
lateral sclerosis.
Description
BACKGROUND OF THE INVENTION
[0001] 1. Field of Invention
[0002] This invention relates to neurotrophic low molecular weight,
small molecule heterocyclic esters and amides having an affinity
for FKBP-type immunophilins, and their use as inhibitors of the
enzyme activity associated with immunophilin proteins, particularly
peptidyl-prolyl isomerase, or rotamase, enzyme activity.
[0003] 2. Description of Related Art
[0004] The term immunophilin refers to a number of proteins that
serve as receptors for the principal immunosuppressant drugs,
cyclosporin A (CsA), FK506 and rapamycin. Known classes of
immunophilins are cyclophilins and FK506 binding proteins, or
FKBPs. Cyclosporin A binds to cyclophilin A while FK506 and
rapamycin bind to FKBP12. These immunophilin-drug complexes
interface with various intracellular signal transduction systems,
especially the immune and nervous systems.
[0005] Immunophilins are known to have peptidyl-prolyl isomerase
(PPIase), or rotamase, enzyme activity. It has been determined that
rotamase enzyme activity plays a role in the catalyzation of the
interconversion of the cis and trans isomers of peptide and protein
substrates for the immunophilin proteins.
[0006] Immunophilins were originally discovered and studied in the
immune tissue. It was initially postulated by those skilled in the
art that inhibition of the immunophilins' rotamase activity leads
to inhibition of T-cell proliferation, thereby causing the
immunosuppressive activity exhibited by immunosuppressant drugs,
such as cyclosporin A, FK506 and rapamycin. Further study has shown
that the inhibition of rotamase activity, in and of itself, does
not result in immunosuppressive activity. Schreiber et al.,
Science, 1990, vol. 250, pp. 556-559. Instead, immunosuppression
appears to stem from the formulation of a complex of
immunosuppressant drugs and immunophilins. It has been shown that
the immunophilin-drug complexes interact with ternary protein
targets as their mode of action. Schreiber et al., Cell, 1991, vol.
66, pp. 807-815. In the case of FKBP-FK506 and cyclophilin-CsA, the
immunophilin-drug complexes bind to the enzyme calcineurin and
inhibit the T-cell receptor signalling which leads to T-cell
proliferation. Similarly, the immunophilin-drug complex of
FKBP-rapamycin interacts with the RAFT1/FRAP protein and inhibits
the IL-2 receptor signalling.
[0007] Immunophilins have been found to be present at high
concentrations in the central nervous system. Immunophilins are
enriched 10-50 times more in the central nervous system than in the
immune system. Within neural tissues, immunophilins appear to
influence nitric oxide synthesis, neurotransmitter release and
neuronal process extension.
[0008] It has been found that picomolar concentrations of an
immunosuppressant such as FK506 and rapamycin stimulate neurite
outgrowth in PC12 cells and sensory neurons, namely dorsal root
ganglion cells (DRGs). Lyons et al., Proc. of Natl. Acad. Sci.,
1994, vol. 91, pp. 3191-3195. In whole animal experiments, FK506
has been shown to stimulate nerve regeneration following facial
nerve injury.
[0009] Surprisingly, it has been found that certain compounds with
a high affinity for FKBPs are potent rotamase inhibitors and
exhibit excellent neurotrophic effects. Furthermore, these rotamase
inhibitors are devoid of immunosuppressive activity. These findings
suggest the use of rotamase inhibitors in creating various
peripheral neuropathies and enhancing neuronal regrowth in the
central nervous system (CNS). Studies have demonstrated that
neurodegenerative disorders such as Alzheimer's disease,
Parkinson's disease, and amyotrophic lateral sclerosis (ALS) may
occur due to the loss, or decreased availability, of a neurotrophic
substance specific for a particular population of neurons affected
in the disorder.
[0010] Several neurotrophic factors affecting specific neuronal
populations in the central nervous system have been identified. For
example, it has been hypothesized that Alzheimer's disease results
from a decrease or loss of nerve growth factor (NGF). It has thus
been proposed to treat SDAT patients with exogenous nerve growth
factor or other neurotrophic proteins, such as brain derived growth
factor, glial derived growth factor, ciliary neurotrophic factor
and neurotropin-3, to increase the survival of degenerating
neuronal populations.
[0011] Clinical application of these proteins in various
neurological disease states is hampered by difficulties in the
delivery and bioavailability of large proteins to nervous system
targets. By contrast, immunosuppressant drugs with neurotrophic
activity are relatively small and display excellent bioavailability
and specificity. However, when administered chronically,
immunosuppressant drugs exhibit a number of potentially serious
side effects including nephrotoxicity, such as impairment of
glomerular filtration and irreversible interstitial fibrosis (Kopp
et al., J. Am. Soc. Nephrol., 1991, 1:162); neurological deficits,
such as involuntary tremors, or non-specific cerebral angina, such
as non-localized headaches (De Groen et al., N. Engl. J. Med.,
1987, 317:861); and vascular hypertension with complications
resulting therefrom (Kahan et al., N. Engl. J. Med., 1989,
321:1725).
[0012] In order to prevent the side access associated with use of
the immunosuppressant compounds, the present invention provides
non-immunosuppressive compounds containing small molecule FKBP
rotamase inhibitors for enhancing neurite outgrowth, and promoting
neuronal growth and regeneration in various neuropathological
situations where neuronal repair can be facilitated, including:
peripheral nerve damage caused by physical injury or disease state
such as diabetes; physical damage to the central nervous system
(spinal cord and brain); brain damage associated with stroke; and
neurological disorders relating to neurodegeneration, such as
Parkinson's disease, SDAT (Alzheimer's disease), and amyotrophic
lateral sclerosis.
SUMMARY OF THE INVENTION
[0013] The present invention relates to neurotrophic low molecular
weight, small molecule compounds having an affinity for FKBP-type
immunophilins. Once bound to these proteins, the neurotrophic
compounds are potent inhibitors of the enzyme activity associated
with immunophilin proteins, particularly peptidyl-prolyl isomerase,
or rotamase, enzyme activity. A key feature of the compounds of the
present invention is that they do not exert any significant
immunosuppressive activity in addition to their neurotrophic
activity.
[0014] Specifically, the present invention relates to a compound of
formula I: 1
[0015] or a pharmaceutically acceptable salt thereof, wherein:
[0016] A and B, together with the nitrogen and carbon atoms to
which they are respectively attached, form a 5-7 membered saturated
or unsaturated heterocyclic ring containing, in-addition to the
nitrogen atom, at least one additional O, S, SO, SO.sub.2, NH or
NR.sub.1 heteroatom in any chemically stable oxidation state;
[0017] X is O or S;
[0018] Z is O, NH or NR.sub.1;
[0019] W and Y are independently O, S, CH.sub.2 or H.sub.2;
[0020] R.sub.1 is C.sub.1-C.sub.6 straight or branched chain alkyl
or alkenyl, which is substituted in one or more position(s) with
(Ar.sub.1).sub.n, (Ar.sub.1).sub.n connected by a C.sub.1-C.sub.6
straight or branched chain alkyl or alkenyl, C.sub.3-C.sub.9
cycloalkyl, C.sub.3-C.sub.9 cycloalkyl connected by a
C.sub.1-C.sub.6 straight or branched chain alkyl or a alkenyl,
Ar.sub.2, or a combination thereof;
[0021] n is 1 or 2;
[0022] R.sub.2 is either C.sub.1-C.sub.9 straight or branched chain
alkyl or alkenyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.5-C.sub.7
cycloalkenyl, or Ar.sub.1,. wherein said alkyl, alkenyl, cycloalkyl
or cycloalkenyl is either unsubstituted or substituted in one or
more position(s) with C.sub.1-C.sub.4 straight or branched chain
alkyl or alkenyl, hydroxyl, or a combination thereof; and
[0023] Ar.sub.1 and Ar.sub.2 are independently a mono-, bi- or
tricyclic, carbo- or heterocyclic ring wherein the ring is either
unsubstituted or substituted in one to three position(s) with halo,
hydroxyl, nitro, trifluoromethyl, C.sub.1-C.sub.6 straight or
branched chain alkyl or alkenyl, C.sub.1-C.sub.4 alkoxy,
C.sub.1-C.sub.4 alkenyloxy, phenoxy, benzyloxy, amino, or a
combination thereof; wherein the individual ring sizes are 5-6
members; and wherein the heterocyclic ring contains 1-6
heteroatom(s) selected from the group consisting of O, N, S, and a
combination thereof.
[0024] A preferred embodiment of this invention is a compound of
formula II: 2
[0025] or a pharmaceutically acceptable salt thereof, wherein:
[0026] A, B and C are independently CH.sub.2, O, S, SO, SO.sub.2,
NH or NR.sub.1;
[0027] R.sub.1 is C.sub.1-C.sub.5 straight or branched chain alkyl
or alkenyl, which is substituted in one or more position(s) with
(Ar.sub.1).sub.n, (Ar.sub.1).sub.n connected by a C.sub.1-C.sub.5
straight or branched chain alkyl or alkenyl, or a combination
thereof;
[0028] n is 1 or 2;
[0029] R.sub.2 is either C.sub.1-C.sub.9 straight or branched chain
alkyl or alkenyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.5-C.sub.7
cycloalkenyl, or Ar.sub.1; and
[0030] Ar.sub.1 is a mono-, bi- or tricyclic, carbo- or
heterocyclic ring, wherein the ring is either unsubstituted or
substituted in one to three position(s) with halo, hydroxyl, nitro,
trifluoromethyl, C.sub.1-C.sub.6 straight or branched chain alkyl
or alkenyl, C.sub.1-C.sub.4 alkoxy, C.sub.1-C.sub.4 alkenyloxy,
phenoxy, benzyloxy, amino, or a combination thereof; wherein the
individual ring sizes are 5-6 members; and wherein the heterocyclic
ring contains 1-6 heteroatom(s) selected from the group consisting
of O, N, S, and a combination thereof.
[0031] Another preferred embodiment is a compound of formula III:
3
[0032] or a pharmaceutically acceptable salt thereof, wherein:
[0033] A, B, C and D are independently CH.sub.2, O, S, SO,
SO.sub.2, NH or NR.sub.1;
[0034] R.sub.1 is C.sub.1-C.sub.5 straight or branched chain alkyl
or alkenyl, which is substituted in one or more position(s) with
(Ar.sub.1).sub.n, (Ar.sub.1).sub.n connected by a C.sub.1-C.sub.6
straight or branched chain alkyl or alkenyl, or a combination
thereof;
[0035] n is 1 or 2;
[0036] R.sub.1 is either C.sub.1-C.sub.9 straight or branched chain
alkyl or alkenyl, C.sub.3-C.sub.9 cycloalkyl, C.sub.5-C.sub.7
cycloalkenyl, or Ar.sub.1; and
[0037] Ar.sub.1 is a mono-, bi- or tricyclic, carbo- or
heterocyclic ring, wherein the ring is either unsubstituted or
substituted in one to three position(s) with halo, hydroxyl, nitro,
trifluoromethyl, C.sub.1-C.sub.6 straight or branched chain alkyl
or alkenyl, C.sub.1-C.sub.4 alkoxy, C.sub.1-C.sub.4 alkenyloxy,
phenoxy, benzyloxy, amino, or a combination thereof; wherein the
individual ring sizes are 5-6 members; and wherein the heterocyclic
ring contains 1-6 heteroatom(s) selected from the group consisting
of O, N, S, and a combination thereof.
[0038] The present invention also relates to a pharmaceutical
composition comprising a neurotrophically effective amount of the
compound of formula I, II or III, and a pharmaceutically acceptable
carrier.
[0039] The present invention further relates to a method of
effecting a neuronal activity in an animal, comprising:
[0040] administering to the animal a neurotrophically effective
amount of the compound of formula I, II or III.
BRIEF DESCRIPTION OF THE DRAWINGS
[0041] FIG. 1(A) is a representative photomicrograph of compound 1
(1 pM) promoting neurite outgrowth in sensory neurons.
[0042] FIG. 1(B) is a representative photomicrograph of compound 1
(10 pM) promoting neurite outgrowth in sensory neurons.
[0043] FIG. 1(C) is a representative photomicrograph of compound 1
(100 pM) promoting neurite outgrowth in sensory neurons.
[0044] FIG. 2(A) is a representative photomicrograph of compound 2
(10 pM) promoting neurite outgrowth in sensory neurons.
[0045] FIG. 2(B) is a representative photomicrograph of compound 2
(100 pM) promoting neurite outgrowth in sensory neurons.
[0046] FIG. 2(C) is representative photomicrograph of compound 2
(10 nM) promoting neurite outgrowth in sensory neurons.
DETAILED DESCRIPTION OF THE INVENTION
Definitions
[0047] "Alkyl" means a branched or unbranched saturated hydrocarbon
chain containing 1 to 6 carbon atoms, such as methyl, ethyl,
propyl, iso-propyl, butyl, iso-butyl, tert-butyl, n-pentyl,
n-hexyl, and the like, unless otherwise indicated.
[0048] "Alkoxy" means the group --OR wherein R is alkyl as herein
defined. Preferably, R is a branched or unbranched saturated
hydrocarbon chain containing 1 to 3 carbon atoms.
[0049] "Halo" means fluoro, chloro, bromo, or iodo, unless
otherwise indicated.
[0050] "Phenyl" includes all possible isomeric phenyl radicals,
optionally monosubstituted or multi-substituted with substituents
selected from the group consisting of alkyl, alkoxy, hydroxy, halo,
and haloalkyl.
[0051] The term "pharmaceutically acceptable salt" refers to salts
of the subject compounds which posses the desired pharmacological
activity and which are neither biologically nor otherwise
undesirable. The salts can be formed with inorganic acids such as
acetate, adipate, alginate, aspartate, benzoate, benzenesulfonate,
bisulfate butyrate, citrate, camphorate, camphorsulfonate,
cyclopentanepropionate, digluconate, dodecylsulfate,
ethanesulfonate, fumarate, glucoheptanoate, glycerophosphate,
hemisulfate heptanoate, hexanoate, hydrochloride hydrobromide,
hydroiodide, 2-hydroxyethanesulfonate, lactate, maleate,
methanesulfonate, 2-naphthalenesulfonate, nicotinate, oxalate,
thiocyanate, tosylate and undecanoate. Base salts include ammonium
salts, alkali metal salts such as sodium and potassium salts,
alkaline earth metal salts such as calcium and magnesium salts,
salt with organic bases such as dicyclohexylamine salts,
N-methyl-D-glucamine, and salts with amino acids such as arginine,
lysine, and so forth. Also, the basic nitrogen-containing groups
can be quarternized within such agents as lower alkyl halides, such
as methyl, ethyl, propyl, and butyl chloride, bromides and iodides;
dialkyl sulfates like dimethyl, diethyl, dibutyl and diamyl
sulfates, long chain halides such as decyl, lauryl, myristyl and
stearyl chlorides, bromides and iodides, aralkyl halides like
benzyl and phenethyl bromides and others. Water or oil-soluble or
dispersible products are thereby obtained.
[0052] The compounds of this invention possess asymmetric centers
and thus can be produced as mixtures of stereoisomers or as
individual stereoisomers. The individual stereoisomers may be
obtained by using an optically active starting material, by
resolving a racemic or non-racemic mixture of an intermediate at
some appropriate stage of the synthesis, or by resolution of the
compound of formula (I). It is understood that the individual
stereoisomers as well as mixtures (racemic and non-racemic) of
stereoisomers are encompassed by the scope of the present
invention. The compounds of this invention possess at least one
asymmetric centers and thus can be produced as mixtures of
stereoisomers or as individual R- and S-stereoisomers. The
individual enantiomers may be obtained by resolving a racemic or
non-racemic mixture of an intermediate at some appropriate stage of
the synthesis. It is understood that the individual R- and S-
stereoisomers as well as mixtures of stereoisomers are encompassed
by this invention. The S-stereoisomer is most preferred due to its
greater activity.
[0053] "Isomers" are different compounds that have the same
molecular formula.
[0054] "Stereoisomers" are isomers that differ only in the way the
atoms are arranged in space.
[0055] "Enantiomers" are a pair of stereoisomers that are
non-superimposable mirror images of each other.
[0056] "Diastereoisomers" are stereoisomers which are not mirror
images of each other.
[0057] "Racemic mixture" means a mixture containing equal parts of
individual enantiomers. "Non-racemic mixture" is a mixture
containing unequal parts of individual enantiomers or
stereoisomers.
[0058] The term "treatment" as used herein covers any treatment of
a disease and/or condition in an animal, particularly a human, and
includes:
[0059] (i) preventing a disease and/or condition from occurring in
a subject which may be predisposed to the disease and/or condition
but has not yet been diagnosed as having it;
[0060] (ii) inhibiting the disease and/or condition, i.e.,
arresting its development; or
[0061] (iii) relieving the disease and/or condition, i.e., causing
regression of the disease and/or condition.
[0062] The system used in naming the compounds of the present
invention is shown below, using a compound of formula II as an
example.
[0063] A compound of formula II wherein A is CH.sub.2, B is S, C is
CH.sub.2, R.sub.1 is 3-phenylpropyl and R.sub.2 is
3,3-dimethylpentyl, is named
3-phenyl-1-propyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-(4-thiazol-
idine) carboxylate.
Compounds of the Invention
[0064] The neurotrophic low molecular weight, small molecule FKBP
inhibitor compounds of this invention have an affinity for
FKBP-type immunophilins, such as FKBP12. When the neurotrophic
compounds of this invention are bound to an FKBP-type immunophilin,
they have been found to inhibit the prolyl-peptidyl cis-trans
isomerase activity, or rotamase, activity of the binding protein
and unexpectedly stimulate neurite growth.
FORMULA I
[0065] In particular, this invention relates to a compound of
formula I: 4
[0066] or a pharmaceutically acceptable salt thereof, wherein:
[0067] A and B, together with the nitrogen and carbon atoms to
which they are respectively attached, form a 5-7 membered saturated
or unsaturated heterocyclic ring containing, in addition to the
nitrogen atom, at least one additional O, S, SO, SO.sub.2, NH or
NR.sub.1 heteroatom in any chemically stable oxidation state;
[0068] X is O or S;
[0069] Z is O, NH or NR.sub.1;
[0070] W and Y are independently O, S, CH.sub.2 or H.sub.2;
[0071] R.sub.1 is C.sub.1-C.sub.6 straight or branched chain alkyl
or alkenyl, which is substituted in one or more position(s) with
(Ar.sub.1).sub.n, (Ar.sub.1).sub.n connected by a C.sub.1-C.sub.6
straight or branched chain alkyl or alkenyl, C.sub.1-C.sub.3
cycloalkyl, C.sub.3-C.sub.9 cycloalkyl connected by a
C.sub.1-C.sub.6 straight or branched chain alkyl or alkenyl,
Ar.sub.2, or a combination thereof;
[0072] n is 1 or 2;
[0073] R.sub.2 is either C.sub.1-C.sub.9 straight or branched chain
alkyl or alkenyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.5-C.sub.7
cycloalkenyl, or Ar.sub.1, wherein said alkyl, alkenyl, cycloalkyl
or cycloalkenyl is either unsubstituted or substituted in one or
more position(s) with C.sub.1-C.sub.4 straight or branched chain
alkyl or alkenyl, hydroxyl, or a combination thereof; and
[0074] Ar.sub.1 and Ar.sub.2 are independently a mono-, bi- or
tricyclic, carbo- or heterocyclic ring, wherein the ring is either
unsubstituted or substituted in one to three position(s) with halo,
hydroxyl, nitro, trifluoromethyl, C.sub.1-C.sub.6 straight or
branched chain alkyl or alkenyl, C.sub.1-C.sub.4 alkoxy,
C.sub.1-C.sub.4 alkenyloxy, phenoxy, benzyloxy, amino, or a
combination thereof; wherein the individual ring sizes are 5-6
members; and wherein the heterocyclic ring contains
1-6heteroatom(s) selected from the group consisting of O, N, S, and
a combination thereof.
[0075] The mono- and bicyclic, carbo- and heterocyclic rings
include without limitation naphthyl, indolyl, furyl, thiazolyl,
thienyl, pyridyl, quinolinyl, isoquinolinyl, fluorenyl and
phenyl.
FORMULA II
[0076] A preferred embodiment of this invention is a compound of
formula II: 5
[0077] or a pharmaceutically acceptable salt thereof, wherein:
[0078] A, B and C are independently CH.sub.2, O, S, SO, SO.sub.2,
NH or NR.sub.1;
[0079] R.sub.1 is C.sub.1-C.sub.6 straight or branched chain alkyl
or alkenyl, which is substituted in one or more position(s) with
(Ar.sub.1).sub.n, (Ar.sub.1).sub.n connected by a C.sub.1-C.sub.6
straight or branched chain alkyl or alkenyl, or a combination
thereof;
[0080] n is 1 or 2;
[0081] R.sub.2 is either C.sub.1-C.sub.9 straight or branched chain
alkyl or alkenyl, C.sub.3-C.sub.9 cycloalkyl, C.sub.5-C.sub.7
cycloalkenyl, or Ar.sub.1; and
[0082] Ar.sub.1 is a mono-, bi- or tricyclic, carbo- or
heterocyclic ring, wherein the ring is either unsubstituted or
substituted in one to three position(s) with halo, hydroxyl, nitro,
trifluoromethyl, C.sub.1-C.sub.6 straight or branched chain alkyl
or alkenyl, C.sub.1-C.sub.4 alkoxy, C.sub.1-C.sub.4 alkenyloxy,
phenoxy, benzyloxy, amino, or a combination thereof; wherein the
individual ring sizes are 5-6 members; and wherein the heterocyclic
ring contains 1-6 heteroatom(s) selected from the group consisting
of O, N, S, and a combination thereof.
[0083] In a particularly preferred embodiment of formula II
compounds:
[0084] A is CH.sub.2;
[0085] B is CH.sub.2 or S;
[0086] C is CH.sub.2 or NH;
[0087] R.sub.1 is selected from the group consisting of
3-phenylpropyl and 3-(3-pyridyl)propyl; and
[0088] R.sub.2 is selected from the group consisting of
3,3-dimethylpentyl, cyclohexyl, and tert-butyl.
[0089] Specific exemplifications of this embodiment are presented
in TABLE I.
1TABLE I No. A B C R.sub.1 R.sub.2 1 CH.sub.2 S CH.sub.2
3-phenylpropyl 3,3-dimethylpentyl 2 CH.sub.2 S CH.sub.2
3-(3-pyridyl)propyl 3,3-dimethylpentyl 3 CH.sub.2 S CH.sub.2
3-phenylpropyl cyclohexyl 4 CH.sub.2 S CH.sub.2 3-phenylpropyl
tert-butyl 5 CH.sub.2 CH.sub.2 NH 3-phenylpropyl 3,3-dimethylpentyl
6 CH.sub.2 CH.sub.2 NH 3-phenylpropyl cyclohexyl 7 CH.sub.2
CH.sub.2 NH 3-phenylpropyl tert-butyl
FORMULA III
[0090] Another preferred embodiment of this invention is a compound
of formula III: 6
[0091] or a pharmaceutically acceptable salt thereof, wherein:
[0092] A, B, C and D are independently CH.sub.2, O, S, SO,
SO.sub.2, NH or NR.sub.1;
[0093] R.sub.1 is C.sub.1-C.sub.5 straight or branched chain alkyl
or alkenyl, which is substituted in one or more position(s) with
(Ar.sub.1).sub.n, (Ar.sub.1).sub.n connected by a C.sub.1-C.sub.6
straight or branched chain alkyl or alkenyl, or a combination
[0094] n is 1 or 2;
[0095] R.sub.1 is either C.sub.1-C.sub.9 straight or branched chain
alkyl or alkenyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.6-C.sub.7
cycloalkenyl, or Ar.sub.1; and
[0096] Ar.sub.1 is a mono-, bi- or tricyclic, carbo- or
heterocyclic ring, wherein the ring is either unsubstituted or
substituted in one to three position(s) with halo, hydroxyl, nitro,
trifluoromethyl, C.sub.1-C.sub.6 straight or branched chain alkyl
or alkenyl, C.sub.1-C.sub.4 alkoxy, C.sub.1-C.sub.4 alkenyloxy,
phenoxy, benzyloxy, amino, or a combination thereof; wherein the
individual ring sizes are 5-6 members; and wherein the heterocyclic
ring contains 1-6 heteroatom(s) selected from the group consisting
of O, N, S, and a combination thereof.
[0097] In a particularly preferred embodiment of formula III
compounds:
[0098] A is CH.sub.2;
[0099] B is CH.sub.2;
[0100] C is S, O or NH;
[0101] D is CH.sub.2;
[0102] R.sub.1 is selected from the group consisting of
3-phenylpropyl and (3,4,5-trimethoxy)phenylpropyl; and
[0103] R.sub.1 is selected from the group consisting of
3,3-dimethylpentyl, cyclohexyl, 3-3-dimethlpropyl, phenyl, and 3,
4, 5-trimethoxyphenyl.
[0104] Specific exemplifications of this embodiment are presented
in TABLE II.
2TABLE II No. A B C D R.sub.1 R.sub.2 8 CH.sub.2 CH.sub.2 S
CH.sub.2 3-phenylpropyl 3,3-dimethylpentyl 9 CH.sub.2 CH.sub.2 O
CH.sub.2 3-phenylpropyl 3,3-dimethylpentyl 10 CH.sub.2 CH.sub.2 S
CH.sub.2 3-phenylpropyl cyclohexyl 11 CH.sub.2 CH.sub.2 O CH.sub.2
3-phenylpropyl cyclohexyl 12 CH.sub.2 CH.sub.2 S CH.sub.2
3-phenylpropyl phenyl 13 CH.sub.2 CH.sub.2 O CH.sub.2
3-phenylpropyl phenyl 14 CH.sub.2 CH.sub.2 NH CH.sub.2
3-phenylpropyl 3,3-dimethylpentyl 15 CH.sub.2 CH.sub.2 NH CH.sub.2
3-phenylpropyl phenyl
[0105] The compounds of the present invention exist as
stereoisomeric forms, either enantiomers or diastereoisomers.
Included within the scope of the invention are the enantiomers, the
racemic form, and diastereoisomeric mixtures. Enantiomers and
diastereoisomers can be separated by methods known to those skilled
in the art.
Methods of Using the Compounds of the Invention
[0106] The compounds of the present invention have an affinity for
the FK506 binding protein, particularly FKBP12, which is present in
the brain. When the inventive compounds bind to FKBP in the brain,
they exhibit excellent neurotrophic activity. This activity is
useful in the stimulation of damaged neurons, the promotion of
neuronal regeneration, the prevention of neurodegeneration, and the
treatment of several neurological disorders known to be associated
with neuronal degeneration and peripheral neuropathies.
[0107] For the foregoing reasons, the present invention further
relates to a method of effecting a neuronal activity in an animal,
comprising:
[0108] administering to the animal a neurotrophically effective
amount of a compound of formula I, II or III.
[0109] In a preferred embodiment, the neuronal activity is selected
from the group consisting of stimulation of damaged neurons,
promotion of neuronal regeneration, prevention of neurodegeneration
and treatment of neurological disorder.
[0110] The neurological disorders that may be treated include but
are not limited to: trigeminal neuralgia; glossopharyngeal
neuralgia; Bell's Palsy; myasthenia gravis; muscular dystrophy;
amyotrophic lateral sclerosis; progressive muscular atrophy;
progressive bulbar inherited muscular atrohpy; herniated; ruptured
or prolapsed invertabrae disk syndromes; cervical spondylosis,
plexus disorders; thoracic outlet destruction syndromes; peripheral
neuropathic such as those caused by lead, dapsone, ticks,
porphyria, or Gullairn-Barr syndrome; Alzheimer's disease; and
Parkinson's disease.
[0111] The compounds of the present invention are particularly
useful for treating a neurological disorder selected from the group
consisting of: peripheral neuropathy caused by physical injury or
disease state, physical damage to the brain, physical damage to the
spinal cord, stroke associated with brain damage, and neurological
disorder relating to neurodegeneration. Examples of neurological
disorders relating to neurodegeneration are Alzheimer's Disease,
Parkinson's Disease, and amyotrophic lateral sclerosis.
[0112] For these purposes the compounds of the present invention
may be administered orally, parenterally, by inhalation spray,
topically, rectally, nasally, buccally, vaginally or via an
implanted reservoir in dosage formulations containing conventional
non-toxic pharmaceutically-acceptable carriers, adjuvants and
vehicles. The term parenteral as used herein includes subcutaneous,
intravenous, intramuscular, intraperitoneally, intrathecally,
intraventricularly, intrasternal and intracranial injection or
infusion techniques.
[0113] To be effective therapeutically as central nervous system
targets, the compounds of the present invention should readily
penetrate the blood-brain barrier when peripherally administered.
Compounds which cannot penetrate the blood-brain barrier can be
effectively administered by an intraventricular route.
[0114] The compounds of the present invention may be administered
in the form of sterile injectable preparations, for example, as
sterile injectable aqueous or oleaginous suspensions. These
suspensions may be formulated according to techniques known in the
art using suitable dispersing or wetting agents and suspending
agents. The sterile injectable preparations may also be sterile
injectable solutions or suspensions in non-toxic
parenterally-acceptable diluents or solvents, for example, as
solutions in 1,3-butanediol. Among she acceptable vehicles and
solvents that may be employed are water, Ringer's solution and
isotonic sodium chloride solution. In addition, sterile, fixed oils
are conventionally employed as solvents or suspending mediums. For
this purpose, any bland fixed oil may be employed including
synthetic mono- or di-glycerides. Fatty acids such as oleic acid
and its glyceride derivatives, including olive oil and castor oil,
especially in their polyoxyethylated versions, are useful in the
preparation of injectables. These oil solutions or suspensions may
also contain long-chain alcohol diluents or dispersants.
[0115] The compounds may be administered orally in the form of
capsules, tablets, aqueous suspensions or solutions. Tablets may
contain carriers such as lactose and corn starch and/or lubricating
agents such as magnesium stearate. Capsules may contain diluents
including lactose and dried corn starch. Aqueous suspensions may
contain emulsifying and suspending agents combined with the active
ingredient. The oral dosage forms may further contain sweetening
and/or flavoring and/or coloring agents.
[0116] The compounds of this invention may also be administered
rectally in the form of suppositories. These compositions can be
prepared by mixing the drug with a suitable non-irritating
excipient which is solid at room temperature, but liquid at rectal
temperature and, therefore, will melt in the rectum to release the
drug. Such materials include cocoa butter, beeswax and polyethylene
glycols.
[0117] The compounds of this invention may also be administered
topically, especially when the conditions addressed for treatment
involve areas or organs readily accessible by topical application,
including neurological disorders of the eye, the skin, or the lower
intestinal tract. Suitable topical formulations are readily
prepared for each of these areas.
[0118] For topical application to the eye, or ophthalmic use, the
compounds can be formulated as micronized suspensions in isotonic,
pH adjusted sterile saline, or, preferably, as solutions in
isotonic, pH adjusted sterile saline, either with or without a
preservative such as benzylalkonium chloride. Alternatively for the
ophthalmic uses the compounds may be formulated in an ointment such
as petrolatum.
[0119] For topical application to the skin, the compounds can be
formulated in a suitable ointment containing the compound suspended
or dissolved in, for example, a mixture with one or more of the
following: mineral oil, liquid petrolatum, white petrolatum,
propylene glycol, polyoxyethylene polyoxypropylene compound,
emulsifying wax and water. Alternatively, the compounds can be
formulated in a suitable lotion or cream containing the active
compound suspended or dissolved in, for example, a mixture of one
or more of the following: mineral oil, sorbitan monostearate,
polysorbate 60, cetyl esters wax, cetearyl alcohol,
2-octyldodecanol, benzyl alcohol and water.
[0120] Topical application for the lower intestinal trace an be
effected in a rectal suppository formulation (see above) or in a
suitable enema formulation.
[0121] Dosage levels on the order of about 0.1 mg to about 10,000
mg of the active ingredient compound are useful in the treatment of
the above conditions, with preferred levels of about 0.1 mg to
about 1,000 mg. The amount of active ingredient that may be
combined with the carrier materials to produce a single dosage form
will vary depending upon the host treated and the particular mode
of administration.
[0122] It is understood, however, that a specific dose level for
any particular patient will depend upon a variety of factors
including the activity of the specific compound employed, the age,
body weight, general health, sex, diet, time of administration,
rate of excretion, drug combination, and the severity of the
particular disease being treated and form of administration.
[0123] The compounds can be administered with other neurotrophic
agents such as neurotrophic growth factor (NGF), glial derived
growth factor, brain derived growth factor, ciliary neurotrophic
factor, and neurotropin-3. The dosage level of other neurotrophic
drugs will depend upon the factors previously stated and the
neurotrophic effectiveness of the drug combination.
Pharmaceutical Compositions of the Invention
[0124] The present invention also relates to a pharmaceutical
composition comprising:
[0125] (i) a neurotrophically effective amount of the compound of
formula I, II or III, and
[0126] (ii) a pharmaceutically acceptable carrier.
[0127] The above discussion relating to the utility and
administration of the compounds of the present invention also
applies to the pharmaceutical compositions of the present
invention.
EXAMPLES
[0128] The following examples are illustrative of the present
invention and are not intended to be limitations thereon. Unless
otherwise specified, all percentages are based on 100% by weight of
the final compound.
Example 1
Synthesis of
3-phenyl-1-propyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-(4-t-
hiazolidine)carboxylate (1)
[0129] 1-(1,2-dioxo-2-methoxyethyl)2-(4-thiazolidine)- carboxylate.
A solution of L-thioproline (1.51 g; 11.34 mmol) in 40 mL of dry
methylene chloride was cooled to 0.degree. C. and treated with 3.3
mL (2.41 g; 23,81 mmol) of triethylamine. After stirring this
mixture for 30 minutes, a solution of methyl oxalyl chloride (1.81
g; 14.74 mmol) was added dropwise. The resulting mixture was
stirred at 0.degree. C. for 1.5 hours, filtered through Celite to
remove solids, dried and concentrated. The crude material was
purified on a silic gel column, eluting with 10% MeOH in methylene
chloride, to obtain 2.0 g of the oxamate as an orange-yellow
solid.
[0130]
3-ohenyl-1-propyl(2S)-1-(1,2-dioxo-2-methoxyethyl)2-(4-thiazolidine-
) carboxylate. 1-
(1,2-dioxo-2-methoxyethyl)2-(4-thiazolidine)carboxylate (500 mg;
2.25 mmol), 3-phenyl-1-propanol (465 mg; 3.42 mmol),
dicyclohexylcarbodiimide (750 mg; 3.65 mmol),
4-dimethylaminopyridine (95 mg; 0.75 mmol) and camphorsulfonic acid
(175 mg; 0.75 mmol) in 30 mL of methylene chloride were stirred
together overnight. The mixture was filtered through Celite to
remove solids and chromatographed (25% ethyl acetate/hexane) to
obtain 690 mg of material, .sup.1H NMR (CDCl.sub.3, 300 MHz):
.delta.1.92-2.01 (m, 2H); 2.61-2.69 (m, 2H); 3.34 (m, 1H);
4.11-4.25 (m, 2H); 4.73 (m, 1H) ; 5.34 (m, 1H); 7.12 (m, 3H) ; 7.23
(m, 2H).
[0131] 3 -phenyl-1-propyl (2 S) -1- (3, 3 -dimethyl- 1,
2-dioxopentyl)-2-(4-thiazolidine) carboxylate (1). A solution of
3-phenyl-1-propyl(2S) -1-(1,2-dioxo-2-methoxyethyl)
2-(4-thiazolidine) carboxylate (670 mg; 1.98 mmol) in
tetrahydrofuran (10 mL) was cooled to -78.degree. C. and treated
with 2.3 mL of a 1.0 M solution of 1, 1-dimethylpropylmagnesium
chloride in ether. After stirring the mixture for 3 hours, it was
poured into saturated ammonium chloride, extracted into ethyl
acetate, and the organic phase was washed with water, dried and
concentrated. The crude material was purified on a silica gel
column, eluting with 25% ethyl acetate in hexane, to obtain 380 mg
of the compound of Example 1 as a yellow oil, .sup.1H NMR
(CDCl.sub.3, 300 MHz): d 0.86 (t, 3H) 1.21 (s, 3H) 1.26 (s, 3H);
1.62-1.91 (m, 3H),; 2.01 (m, 2H) ; 2.71 (m, 2H) ; 3.26-3.33 (m, 2H)
; 4.19 (m, 2H); 4.58 (m, 1H); 7.19 (m, 3H); 7.30 (m, 2H). Anal.
Clcd. for C.sub.20H.sub.27NO.sub.4S: C, 63.63; H, 7.23; N, 3.71.
Found: C, 64.29; H, 7.39; N, 3.46.
Example 2
Synthesis of
3-(3-pyridyl)-1-propyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)
-2-(4-thiazolidine) carboxylate (2)
[0132] The compound of Example 2 was prepared according to the
procedure of Example 1, using 3-(3-pyridyl)-1-propanol in the final
step, to yield 3-
(3-pyridyl)-1-propyl(2S)-1-(3,3-dimethyl-1,2-dioxopentyl)-2-(4-thiazol-
idine) carboxylate, .sup.1H NMR (CDCl.sub.3, 300 MHz): .epsilon.
0.89 (t, 3H, J=7.3); 1.25 (s, 3H); 1.28 (s, 3H) ; 1.77 (q, 2H,
J=7.3); 2.03 (t, 2, J=6.4, 7.5); 2.72 (t, 2H, J=7.5); 3.20 (dd, 1H,
J=4.0, 11.8); 3.23 (dd, 1 H, J=7.0, 11.8); 4.23 (t, 2H, J=6.4);
4.55 (d, 2H, J=8.9); 5.08 (dd, 1H, J=4.0, 7.0); 7.24 (m, 1H); 8.48
(m, 2H). Anal. Caldc. for C.sub.13H.sub.26N.sub.2O.sub.4S-0.5
H.sub.2O: C, 58.89; H 7.02; N, 7.23. Found: C, 58.83; H:, 7.05; N
7.19.
[0133] As discussed above, the compounds of the present invention
have an affinity, for the FK506 binding protein, particularly
FKBP12. The inhibition of the prolyl peptidyl cis-trans isomerase
activity of FKBP may be measured as an indicator of this
affinity.
Ki Test Procedure
[0134] Inhibition of the peptidyl-prolyl isomerase (rotamase)
activity of the inventive compounds can be evaluated by known
methods described in the literature (Harding, et al., Nature, 1989,
341:758-760; Holt et al. J. Am. Chem. Soc., 115:9923-9938). These
values are obtained as apparent Ki's and are presented in Table
III. The cis-trans isomerization of an alanine-proline bond in a
model substrate, N-succinyl-Ala-Ala-Pro-Phe-p-n- itroanilide, is
monitored spectrophotometrically in a chymotrypsin-coupled assay,
which releases paranitroanilide from the trans form of the
substrate. The inhibition of this reaction caused by the addition
of different concentrations of inhibitor is determined, and the
data is analyzed as a change in first-order rate constant as a
function of inhibitor concentration to yield the apparent Ki
values.
[0135] In a plastic cuvette are added 950 mL of ice cold assay
buffer (25 mM HEPES, ph 7.8, 100 mM NaCl), 10 mL of FKBP (2.5 mM in
10 mM Tris-Cl ph 7.5, 100 mM NaCl, 1 mM dithiothreitol), 25 mL of
chymotrypsin (50 mg/ml in 1 mM HCl) and 10 mL of test compound at
various concentrations in dimethyl sulfoxide. The reaction is
initiated by the addition of 5 mL of substrate
(succinyl-Ala-Phe-Pro-Phe-para-nitroanilide, 5 mg/mL in 2.35 mM
LiCl in trifluoroethanol).
[0136] The absorbance at 390 nm versus time is monitored for 90
seconds using a spectrophotometer and the rate constants are
determined from the absorbance versus time data files.
[0137] The data for these experiments for representatives compounds
are presented in Table III under the column "Ki".
[0138] The neurotrophic effects of the compounds of the present
invention can be demonstrated in cellular biological experiments in
vitro, as described below.
Chick Dorsal Root Ganglion
Cultures and Neurite Outgrowth
[0139] Dorsal root ganglia were dissected from chick embryos of ten
day gestation. Whole ganglion explants were cultured on thin layer
Matrigel-coated 12 well plates with Liebovitz L15 plus high glucose
media supplemented with 2 mM glutamine and 10% fetal calf serum,
and also containing 10 .mu.M cytosine .beta.-D arabinofuranoside
(Ara C) at 37.degree. C. in an environment containing 5% CO.sub.2.
Twenty four hours later, the DRGs were treated with various
immunophilin ligands. Forty-eight hours after drug treatment, the
ganglia were visualized under phase contrast or Hoffman Modulation
contrast with a Zeiss Axiovert inverted microscope.
Photomicrographs of the explants were made, and neurite outgrowth
was quantitated. Neurites longer than the DRG diameter were counted
as positive, with total number of neurites quanitated per each
experimental condition. Three to four DRGs are cultured per well,
and each treatment was performed in duplicate.
[0140] The data for these experiments for representative compounds
are presented in the "ED50" column of Table III. Representative
photomicrographs of compounds 1 (1 pM, 10 pM, 100 pM) and 2 (10 pM,
100 pM, 10 nM) promoting neurite outgrowth in sensory neurons are
shown in FIGS. 1(A-C) and 2(A-C), respectively.
3TABLE III In Vitro Test Results Compound Ki.nM ED60 nM 1 215 0.031
2 638 2.0
[0141] All publications and patents identified above are hereby
incorporated by reference.
[0142] The invention being thus described, it will be obvious that
the same may be varied in many ways. Such variations are not to be
regarded as a departure from the spirit and scope of the invention
and all such modifications are intended to be included within the
scope of the following claims.
* * * * *