Use of K-252A derivative for treatment of peripheral or central nerve disorders, and cytokine overproduction

Abstract

Disclosed herein are therapeutic methodologies utilizing a ring substituted derivative of the indolocarbazole K252a, the derivative represented by the formula (A). The compound is useful for treating peripheral neuropathies, central neuronal degeneration and cytokine overproduction. Typical diseases related to the above are peripheral neuropathy, Alzheimer's disease, Parkinson's disease and autoimmune and allergic conditions.

Description

[0001] Field

[0002] The present invention relates to ring substituted derivatives of K-252a for use in methods of alleviating the deleterious effects of various diseases, disorders and disorders. Background of the invention

[0003] 1. Indolocarbazole K-252a

[0004] K-252a is a compound containing an indolocarbazole skeleton [Japanese Published Unexamined Patent Application 41489 / 85 (US 4555402)], which has a stereochemistry as shown in Formula I:

[0005] It has been reported that K-252a very potently inhibits protein kinase C (PKC), which plays an important role in the regulation of cellular functions, and has various activities such as inhibition of smooth muscle contraction (Jpn. J Pharmacol. 43 (Suppl.): 284, 1987), the effect of inhibiting the secretion of serotonin (Biochem. Biophys. Res. Commun.) 144: 35, 1987), the effect of inhibiting the elongation of Science Journal (J.Neuroscience), 8: 715, 1988), the effect of inhibiting the release of histamine (Allergy (...

Examples

Embodiment 1

[0064] Pharmacological activity on dopaminergic neurons

[0065] Experiments with Compound A were performed on MPTP-injured dopaminergic neurons in mice (MPTP mouse model). Subcutaneous administration of a single dose of MPTP (20 mg / kg) to C57 black mice resulted in a loss of approximately 60% of striatal tyrosine hydroxylase activity. Daily administration of 0.01-10 mg / kg (sc) of Compound A to the MPTP-treated mice reduced the loss of striatal tyrosine hydroxylase activity. These data are listed in Table 2.

[0066] Table 2

[0067] deal with

striatal TH activity

(±SEM)

% of uninjured control

undamaged

MPTP-impaired

MPTP-lesioned (vector)

MPTP-impaired + Compound A

19.06±1.06

7.40±1.07

6.99±0.75

100±5.6

39±5.6

37±4.0

(0.01mg / kg)

(0.03mg / kg)

(0.10mg / kg)

(0.30mg / kg)

(1.0mg / kg)

(3.0mg / kg)

(10.0mg / kg)

7.76±1.23

10.48±0.92 *

11.51±0.80 *

...

Embodiment 2

[0078] Pharmacological activity on GABAergic neurons

[0079] Compound A was tested for its ability to inhibit the loss of GABAergic neurons (eg, enhance survival) in the macrocellular inferior olivary nucleus using the well-known ibotenic acid injury model. Ibotenic acid, an excitotoxin, is known to reduce the number of GABA-expressing neurons in the nbm region (Lindefors et al., Neurosci. Lett., 135:262-264, 1992; Shaugnessy et al. Brain Res., 637 : 15-26, 1994).

[0080] Ibotenic acid (5.0 g) was injected into the side of the nbm of adult male Sprague-Dawley rats. Eighteen hours after injury, administration of compound A (0.03 mg / kg) to rats was started by subcutaneous injection, every other day, and continued until 18 days after injury. Tissue sections of the whole nbm rostral-caudalextent were then collected and processed for detection of glutamate decarboxylase, an enzyme required for the biosynthesis of GABA. The number of glutamate decarboxylase-expressing neurons w...

Embodiment 3

[0084] Pharmacological activity on NBM neurons

[0085]To directly test the ability of Compound A to prevent excitatory neuronal death, nbm neurons in adult Sprague-Dawley rats were first labeled with a long-lived marker. The labeling is done by injecting Fluoro-Gold (FG), a protein that is absorbed by nerve endings and transported back to cells (Book et al., J. Neuropath. Exp. Neurology, 53:95-102, 1994), before entering and Neuronal tracers of NBM neuronal targets within the parietal cortex. After 7-10 days, animals received unilateral NBM injury with 5 μg ibotenic acid. Beginning 18 hours after the injury, the animals were subcutaneously injected with Compound A (0.03 mg / kg) every other day until 18 days after the injury. Tissue sections of the entire anterior-caudal region of the nbm on both sides of the brain were collected, and the number of nbm neurons labeled with FG was counted. Count values ​​were corrected for magnitude differences using standard methods and re...

[0001] Field

[0002] The present invention relates to ring substituted derivatives of K-252a for use in methods of alleviating the deleterious effects of various diseases, disorders and disorders. Background of the invention

[0003] 1. Indolocarbazole K-252a

[0004] K-252a is a compound containing an indolocarbazole skeleton [Japanese Published Unexamined Patent Application 41489 / 85 (US 4555402)], which has a stereochemistry as shown in Formula I:

[0005] It has been reported that K-252a very potently inhibits protein kinase C (PKC), which plays an important role in the regulation of cellular functions, and has various activities such as inhibition of smooth muscle contraction (Jpn. J Pharmacol. 43 (Suppl.): 284, 1987), the effect of inhibiting the secretion of serotonin (Biochem. Biophys. Res. Commun.) 144: 35, 1987), the effect of inhibiting the elongation of Science Journal (J.Neuroscience), 8: 715, 1988), the effect of inhibiting the release of histamine (Allergy (...

Examples

Embodiment 1

[0064] Pharmacological activity on dopaminergic neurons

[0065] Experiments with Compound A were performed on MPTP-injured dopaminergic neurons in mice (MPTP mouse model). Subcutaneous administration of a single dose of MPTP (20 mg / kg) to C57 black mice resulted in a loss of approximately 60% of striatal tyrosine hydroxylase activity. Daily administration of 0.01-10 mg / kg (sc) of Compound A to the MPTP-treated mice reduced the loss of striatal tyrosine hydroxylase activity. These data are listed in Table 2.

[0066] Table 2

[0067] deal with

striatal TH activity

(±SEM)

% of uninjured control

undamaged

MPTP-impaired

MPTP-lesioned (vector)

MPTP-impaired + Compound A

19.06±1.06

7.40±1.07

6.99±0.75

100±5.6

39±5.6

37±4.0

(0.01mg / kg)

(0.03mg / kg)

(0.10mg / kg)

(0.30mg / kg)

(1.0mg / kg)

(3.0mg / kg)

(10.0mg / kg)

7.76±1.23

10.48±0.92 *

11.51±0.80 *

...

Embodiment 2

[0078] Pharmacological activity on GABAergic neurons

[0079] Compound A was tested for its ability to inhibit the loss of GABAergic neurons (eg, enhance survival) in the macrocellular inferior olivary nucleus using the well-known ibotenic acid injury model. Ibotenic acid, an excitotoxin, is known to reduce the number of GABA-expressing neurons in the nbm region (Lindefors et al., Neurosci. Lett., 135:262-264, 1992; Shaugnessy et al. Brain Res., 637 : 15-26, 1994).

[0080] Ibotenic acid (5.0 g) was injected into the side of the nbm of adult male Sprague-Dawley rats. Eighteen hours after injury, administration of compound A (0.03 mg / kg) to rats was started by subcutaneous injection, every other day, and continued until 18 days after injury. Tissue sections of the whole nbm rostral-caudalextent were then collected and processed for detection of glutamate decarboxylase, an enzyme required for the biosynthesis of GABA. The number of glutamate decarboxylase-expressing neurons w...

Embodiment 3

[0084] Pharmacological activity on NBM neurons

[0085]To directly test the ability of Compound A to prevent excitatory neuronal death, nbm neurons in adult Sprague-Dawley rats were first labeled with a long-lived marker. The labeling is done by injecting Fluoro-Gold (FG), a protein that is absorbed by nerve endings and transported back to cells (Book et al., J. Neuropath. Exp. Neurology, 53:95-102, 1994), before entering and Neuronal tracers of NBM neuronal targets within the parietal cortex. After 7-10 days, animals received unilateral NBM injury with 5 μg ibotenic acid. Beginning 18 hours after the injury, the animals were subcutaneously injected with Compound A (0.03 mg / kg) every other day until 18 days after the injury. Tissue sections of the entire anterior-caudal region of the nbm on both sides of the brain were collected, and the number of nbm neurons labeled with FG was counted. Count values ​​were corrected for magnitude differences using standard methods and re...

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