DE102007001804A1 - Use of GABAB receptor selective agonists for the treatment of inflammatory skin diseases - Google Patents

Abstract

The invention relates to an application of GABA <SUB> B </ SUB> receptor selective agonists, in particular the GABA <SUB> B </ SUB> receptor agonist baclofen in the treatment of inflammatory skin diseases.

Description

Background of the invention

The present invention relates to the use of GABA _B receptor selective agonists for the treatment of inflammatory skin diseases.

GABA receptors are transmembrane proteins that mainly occur in the central nervous system (CNS) and peripheral nervous system in nerve cells, but also in other cell types, and specifically bind the neurotransmitter γ-aminobutyric acid (GABA). Studies strongly suggest that GABA mediates the inhibitory effects of local interneurons in the brain as well as presynaptic inhibitory effects within the spinal cord. In contrast to the GABA _A and GABA _C receptors, which form ion channels, the metabotropic GABA _B receptor belongs to the family of G protein-coupled receptors. The metabotropic GABA _B receptor was first described in 1996 in a patent application Novartis ( US 020020091250A1 ) and is considered to be one of the most important inhibitory receptors of synaptic transmission. In addition to being the major central nervous system, the GABA _B receptor is also expressed in the lungs, liver, gut, and immune system cells. While the function of the GABA _B receptor in the nervous system or in the digestive system has already been very well researched, its role in the immune system is largely unknown. Accordingly, (Ant) agonists of the GABA _B receptor find application in the treatment and study of diseases of the nervous system, such as epilepsy, Parkinson, multiple sclerosis, anxiety disorders and depression, as well as disorders of the digestive system, such as reflux oesophagitis and dyspepsia ( Overview in Bettler et al. 2004 and Cryan & Kaupmann, 2005 ).

Baclofen, β-p-chlorophenyl-GABA, is considered a prototype of the GABA _B receptor agonists and is currently used mainly for its muscle-relaxing and analgesic properties. Baclofen has been in the 70 years to treat spastic disorders among drug addicts used and is now under the brand name Lioresal ^® (Fa. Novartis) for the treatment of spasticity in spinal cord injury and multiple sclerosis. In addition, baclofen is mainly used as a muscle relaxant. Other previously known applications relate to the treatment of the central nervous system (CNS), the digestive system, lung, kidney and bile.

lupus Erythematosus is a collective name for autoimmune diseases the skin and internal organs. There are 3 different forms: 1. Skin manifestation, 2. systemic involvement, u. a. SLE, and 3. neonatal syndromes. In the case of SLE are different Patterns of organ involvement, with arthritis (85%), skin symptoms (50-60%), blood count changes (60%), lupus nephritis (51%), pleurisy (56%), endocarditis, neurol. and psych. disorders (23-30%) can occur.

Hosseinzadeh H, Ramezani M, Fadishei M, and Mahmoudi M. (Antinociceptive, antiinflammatory and acute toxicity effects of Zhumeria majdae extracts in mice and rats., Phytomedicine 2002 Mar; 9 (2): 135-41) compare the antinociceptive and antiinflammatory effects of an ethanolic plant extract of the Iranian medicinal plant Zhumeria majdae with the effects of baclofen or dexamethasone in acute or chronic inflammation. The chronic inflammatory test is used in mice and rats. As a result, the efficiency of the plant extract in chronic inflammation is comparable to the efficiency of baclofen.

Lu Y and Westlund KN. (Effects of baclofen on colon inflammation-induced Fos, CGRP and SP expression in spinal cord and brainstem., Brain Res. 2001 Jan 19; 889 (1-2): 118-30) investigate the acute inflammation caused by mustard oil in the lumen of the intestine of rats and the associated expression of the proto-oncogene Fos in neurons of the brainstem and the spine. Systemic administration of baclofen not only significantly reduces Fos expression in the spine after mustard oil treatment, but also reduces immunoreactivity in neurons of the spine, relative to the elevated levels that have formed after intestinal inflammation. This shows a significant reducing influence of baclofen on inflammatory processes.

Song DK, Suh HW, Huh SO, Jung JS, Him BM, Choi IG, and Kim YH. (Central GABAA and GABAB receptor modulation of basal and stress-induced plasma interleukin-6 levels in mice J Pharmacol Exp Ther. 1998 Oct; 287 (1): 144-9) investigate the modulatory role of GABA _A and GABA _B receptors in the regulation of basal and stress - induced levels of the cytokine interleukin - 6 (IL - 6). Cytokines such as IL-6 are increasingly found in inflammatory processes. The studies in mice showed that baclofen's increase of stress-induced IL-6 could prevent plasma levels. It is pointed to a suppressive modulation of the GABA receptors via their agonists.

Taylor Gjevre RM, and Gjevre YES. (Anti-glutamic acid decarboxylase antibodies in a patient with systemic lupus erythematosus and fibromyalgia symptoms., Lupus. 2005; 14 (6): 486-8) describe the effect of baclofen in the treatment of a patient with the existing skin disease systemic lupus erythematosus. Mention is made of the beneficial effect of baclofen on the onset of myalgic pain symptoms. A connection between the effect of baclofen and the immunological symptoms of this autoimmune disease is suspected.

US2005226927 claims and discloses pharmaceutical dosage forms for the immediate and controlled release of GABA _B receptor agonists such as baclofen. However, the application does not refer to inflammation of the skin.

Because of incomplete knowledge of immunopathogenesis occurs the treatment of inflammatory skin diseases often empirically. Therapeutic measures that take into account the cause are usually not possible. The established systemic Although therapies can lead to remission of the disease. However, there may be a high potential for side effects be accepted. For chronic inflammatory diseases, Like rheumatoid arthritis, there are first experiences with one cytokine modulating therapy. By progress z. B. in the understanding the immunopathophysiology of psoriasis as a T-cell mediated Dermatosis, the use of innovative drugs as one further, more specific and effective treatment option considered this and other inflammatory skin diseases become. The immunological treatment strategies are aimed in particular on regulation of T-cell activation, inhibition of adhesion inflammatory cells, inhibiting the action of proinflammatory Mediators and on the use of anti-inflammatory cytokines. There are a number of skin conditions caused by UV light therapy (UV = ultraviolet radiation) can be treated. Both long-wave UVA and UVB rays are used. In recent years, the high-dose UVA-1 therapy as a new phototherapeutic principle for the treatment of inflammatory Skin diseases, especially atopic dermatitis, introduced become.

The The main object of the present invention is the improvement and simplification of existing treatment options of inflammatory skin diseases such. Psoriasis, Autoimmune diseases or wound healing. With the previously used Means could be a quick and thorough treatment of the before so far not achieved.

The object of the present invention is achieved by the use of GABA _B receptor selective agonists such. B. the GABA _B receptor agonist baclofen by the preparation of a pharmaceutical composition for the treatment of inflammatory skin diseases.

Summary of the invention

The invention relates to an application of GABA _B receptor selective agonists, in particular the GABA _B receptor agonist baclofen in the treatment of inflammatory skin diseases.

Brief description of the figures

1 : The administration of baclofen (200 μl, 1 mg / kg) causes a significant swelling of the irritated ear

2 : Exemplary comparison of a treated and untreated mouse ear

3 : A triple staining peak (nuclei), CD31 (endothelial cells), CD45 (leukocytes)

The Figure shows that in the treated sample despite existing Blood vessels restricted the infiltration of immune cells is.

4 : Dose-response dependence of treatment with baclofen

Detailed description the invention

The invention relates to an application of a GABA _B receptor selective agonist, z. B. baclofen, for the treatment of inflammatory skin diseases.

The object of the present invention is achieved by using a GABA _B receptor selective agonist for the treatment of inflammatory skin diseases.

The objects of the invention are also achieved by a method of treating inflammatory skin diseases, comprising administering a GABA _B receptor selective agonist to a patient having an inflammatory skin disease.

Any GABA _B receptor selective agonist may be used in the present invention. Examples of suitable GABA _B receptor selective agonists include baclofen, 3- (aminopropyl) methyl phosphinic acid and its salts, 3-aminopropyl phosphinic acid and gabapentin (1- (aminomethyl) cyclohexylacetic acid).

In one embodiment, the GABA _B receptor is a selective agonist selected from the group of full agonists and partial agonists and the allosteric modulator group.

The term "GABA _B receptor selective agonist" refers to a substance capable of producing a similar effect to the physiological ligand at the GABA _B receptor. "GABA _B receptor selective agonists" include the "full agonists" which " partial agonists "and the" allosteric modulators ".

"Full Agonists "are substances that produce maximum effect, while "partial agonists" are substances, which cause a submaximal effect.

"Allosteric modulators" alter (modulate) the spatial structure (quaternary structure) of a receptor such that binding of one ligand has a different effect: Previously known allosteric modulators such as CGP 7930 and CGP 13501 bind to the transmembrane region of GABA _B Receptor and stabilize the receptor in its active form, resulting in a higher affinity for agonists and a higher efficacy Further allosteric modulators currently under development bind to the so-called GB ₂ subunit of the GABA _B receptor.

In In another embodiment, the full agonist is baclofen or gabapentin.

Baclofen is present in compounds as the (±) form of baclofen, (+) - form or as (-) - form. The (-) form of baclofen is generally preferred because studies have shown that this form is even more selective for GABA _B receptors.

• CGP44532 = (S)-(–)

In one embodiment, CGP 7930, GS 39783, CGP 44532 or CGP 13501 is the allosteric modulator.

• CGP44532 = (S) - (-)

The chemical name of the allosteric modulators identified herein by their structural formula is CGP 7930 [2,6-di-tert-butyl-4- (3-hydroxy-2,2-dimethyl-propyl) -phenol], CGP 13501 Aldehyde analogue thereof, for GS 39783 is N, N'-dicyclopentyl-2-methylsulfanyl-5-nitro-pyrimidine-4,6-diamine ( Bettler et al., Physiol. Rev. 2004, p. 857 ) and is for CGP 44532 (3-amino-2 [S] -hydroxypropyl) -methylphosphinic acid ( Bowery et al., Pharm. Rev. 2002, p. 254 ).

In One embodiment is the inflammatory skin disease selected from the group of erythematosquamous Dermatoses, autoimmune diseases, eczema, allergies, Infectious diseases of the skin and wound healing.

In One embodiment is the erythematosquamous Dermatosis psoriasis.

In In another embodiment, the autoimmune diseases a blistering autoimmune dermatosis or collagenosis, especially lupus erythematosus.

In In one embodiment, the eczema diseases are eczema or allergic contact dermatitis.

In In one embodiment, the infectious diseases the skin caused by viruses, bacteria, fungi or parasites.

In one embodiment, the GABA _B receptor selective agonist is administered to a patient having an inflammatory skin disease as defined above.

In one embodiment, the GABA _B receptor is administered selective agonist, preferably baclofen, at a dose of about 3 μg / kg body weight / day to about 5 mg / kg body weight / day.

In another embodiment, the GABA _B receptor is administered selective agonist, preferably baclofen, at a dose of from about 10 μg / kg body weight / day to about 2.5 mg / kg body weight / day.

Determination of the appropriate dose of GABA _B receptor selective agonist for a particular use is at the discretion of the skilled artisan. Usually, treatment is initiated with a smaller dose that is less than the optimal dose of the substance. Subsequently, the dose is increased in small increments until the optimal effect is achieved under the conditions. The amount and frequency of administration of the GABA _B receptor selective agonist will be at the discretion of the treating physician, taking into account the following factors such as age, physical condition and size of the patient as well the severity of the disease to be treated. Children generally start with a dose of about 25 μg per child per day.

In one embodiment, the GABA _B receptor is administered selective agonist in a pharmaceutical composition.

In An embodiment comprises the pharmaceutical composition a pharmaceutically acceptable carrier.

The pharmaceutical compositions of the present invention with conventional pharmaceutically acceptable carriers such as excipients and additives using conventional techniques getting produced. Such pharmaceutically acceptable carriers include binders, flavorings, buffers, thickeners, dyes, Dispersants, suspending agents, fragrances, preservatives, lubricants and the same.

In one embodiment, administration of the GABA _B receptor selective agonist is local or systemic.

In one embodiment, administration of the GABA _B receptor selective agonist is orally, subcutaneously, intracutaneously, parenterally, transdermally, topically, intravenously, intraarterially, intramuscularly, intraperitoneally, or in a combination of the foregoing ways.

The Administration of the pharmaceutical composition may be oral, subcutaneous, intracutaneous, parenteral, transdermal, topical, intravenous, intraarterially, intramuscularly, intraperitoneally or through Any other acceptable method is done.

The Pharmaceutical composition of the present invention can be administered in any type of conventional dosage forms. Solid dosage forms include capsules, tablets, pills, powders, Suspensions, solutions or suppositories.

parenteral Dosage forms include sterile solutions or suspensions. Inhalation administration may take the form of an oral spray or done by blowing.

In In one embodiment, the administration is transdermally or topically.

In In another embodiment, the administration takes place Transdermal In one embodiment, the transdermal Administration in a dosage form selected from the group consisting of spray, plaster, ointment, semi-solid cream, Liquid like a solution, solid like a Powder, gel, emulsion or suspension.

transdermal Dosage forms of the conventional reservoir or matrix plaster type and the like can also be applied.

Another embodiment is a GABA _B receptor selective agonist as defined above for the treatment of inflammatory skin diseases as defined above.

The inventors were able to show for the first time that GABA _B receptor selective agonists such. As baclofen, in addition to their known effects have an anti-inflammatory effect in inflammatory skin diseases and therefore can be successfully used for their treatments. In particular, the inventors were able to show for the first time that GABA _B receptor selective agonists such. As baclofen, in an established inflammatory model in the mouse can exert an anti-inflammatory effect. The results presented in the accompanying figures show that in the so-called "ear swelling response" model, ear swelling induced by DNFB is almost completely suppressed by the administration of baclofen.

Further control experiments showed that up to a dose of 10 ^-5 M, no toxic influence of baclofen on lymphocytes and monocytes (PBMC) or leucocytes is detectable.

The mechanism of the surprisingly significant effect is unknown and is currently under investigation. Without wishing to be bound by any particular mechanism of action, one possible mechanism of action is a change in expression induced by the GABA _B receptor selective agonist of adhesion molecules that regulate the extravasation of immune cells in the skin. In addition, down-regulation of chemokine receptors involved in "signaling" immune cells into the skin or a reduction in the production of inflammatory cytokines could play a role.

in the The following will be referred to the examples which illustrate the invention by way of example, without them in any way limit.

Example 1:

Baclofen reduces the inflammation in the "ear swelling response" model

Around the present mouse model for inflammation, the so-called "ear swelling response" model, on the effect of baclofen, the ears of C57BL / 6 mice were subjected to the following procedure over a period of 6 days:

Day 1: Sensitization phase:

The Mice were sensitized by DNFB as a sensitizing solution (0.025 g of DNFB in a 5 ml acetone-olive oil mixture). 75 μl were distributed on the back of the animals.

Day 5: Re-exposure phase:

The Mice received DNFB solution as a re-exposure solution (0.015 g DNFB in a 5 ml acetone-olive oil mixture) only on an ear.

Immediately afterwards
Group 1 with 200 μl baclofen (1 mg / kg) injected intraperitoneally,
Group 2 with 200 ul PBS intraperitoneally injected.

Day 6

The Mice were sacrificed and ear thickness was measured determined a caliper. The thicker the ear, the better higher is the degree of ignition.

A Total number of 15 C57BL / 6 mice was used per group.

As a result, the DNFB-induced ear swelling reaction in C57BL / 6 mice was significantly reduced by treatment with baclofen versus mock treatment with PBS, see 1 , The reduction in inflammation almost reached the ear thickness of untreated C57BL / 6 mice.

Example 2:

Baclofen reduces inflammation in the "Ear Swelling Response" model

Around Giemsa stained tissue sections of the treated To get inflamed ears were 2-3 glass slides incubated with the tissue sections from the ear in Grunewald's solution. Then the glass slides were washed with water. Subsequently The glass slides were diluted 1:10 with a Giemsa solution incubated for 20 minutes. After that the glass slides were washed again with water. At the end were the glass slides are covered with a fixing medium.

In 2 It can be clearly seen that the tissue sections stained with Giemsa have a clearly recognizable reduced ear swelling after baclofen treatment in comparison with Giemsa-stained untreated tissue sections.

Example 3:

Baclofen treatment leads to reduced infiltration of leukocytes

Immunofluorescence staining of frozen tissue sections.

• 1. Das Gewebe wurde in 4% Paraformaldehyd für 10 Minuten bei Raumtemperatur fixiert.
• 2. Das Gewebe wurde 5 Minuten in PBS gewaschen.
• 3. Das Gewebe wurde in Blockierungslösung (5% Ziegenserum, 0.1% Triton X100 und 1% BSA in PBS-Lösung) inkubiert.
• 4. Dann wurde für 1 Stunde bei Raumtemperatur mit primärem Antikörper in PBS-1% BSA (1:200 verdünnt) inkubiert.
• 5. Danach wurde zweimal mit PBS gewaschen.
• 6. Die Glasträger wurden mit sekundärem Antikörper, gekoppelt an Fluoreszenzfarbstoff (1:1000) in PBS-1% BSA, im Dunklen für 45 Minuten inkubiert (1:1000 verdünnt).
• 7. Danach wurde zweimal mit PBS gewaschen.
• 8. Es wurde in Hoechst-Lösung (1:1000 verdünnt) für 5 Minuten inkubiert.
• 9. Die Glasträger wurden in PBS gewaschen.
• 10. Die Glasträger wurden dann mit Aqua-Polymount bedeckt.
• 11. Die Färbung konnte danach mit Hilfe eines Fluoreszenzmikroskopes sichtbar gemacht werden.

The ears of the examined mice were frozen immediately after dissection of the tissue at -80 ° C in "Tissue-Tech" solution (Sakwa, Cryomold Standard), then the following procedure was performed:

• 1. The tissue was fixed in 4% paraformaldehyde for 10 minutes at room temperature.
• 2. The tissue was washed in PBS for 5 minutes.
• 3. The tissue was incubated in blocking solution (5% goat serum, 0.1% Triton X100 and 1% BSA in PBS solution).
• 4. Then incubate for 1 hour at room temperature with primary antibody in PBS-1% BSA (diluted 1: 200).
• 5. Thereafter, washed twice with PBS.
• 6. Glass slides were incubated with secondary antibody coupled to fluorescent dye (1: 1000) in PBS-1% BSA in the dark for 45 minutes (diluted 1: 1000).
• 7. Thereafter, washed twice with PBS.
• 8. It was incubated in Hoechst's solution (diluted 1: 1000) for 5 minutes.
• 9. The glass slides were washed in PBS.
• 10. The glass slides were then covered with Aqua-Polymount.
• 11. The staining could then be visualized using a fluorescence microscope.

As in 3 clarified, the baclofen treatment leads in the investigated mice to reduced infiltration of leucocytes, which is reflected in the immunofluorescent staining in a reduced ear swelling.

Example 4:

Baclofen reduces the inflammation in the "ear swelling response" model, dose-response curve

Around the influence of different baclofen concentrations on the Ear inflammation was investigated using various baclofen Concentrations between 0.01 and 1 mg / kg used.

The protocol used for this purpose is described in Example 1.

4 illustrates the dose-response relationship of various baclofen concentrations used on the inflammation of the ear.

The in the above description, the claims and Features of the invention disclosed in the drawings both individually and in any combination for the Realization of the invention in its various embodiments be essential.

QUOTES INCLUDE IN THE DESCRIPTION

This list The documents listed by the applicant have been automated generated and is solely for better information recorded by the reader. The list is not part of the German Patent or utility model application. The DPMA takes over no liability for any errors or omissions.

Cited patent literature

• - US 020020091250 A1 [0002]
• US 2005226927 [0009]

Cited non-patent literature

• - Overview in Bettler et al. 2004 and Cryan & Kaupmann, 2005 [0002]
• - Hosseinzadeh H, Ramezani M, Fadishei M, and Mahmoudi M. (Antinociceptive, antiinflammatory and acute toxicity effects of Zhumeria majdae extracts in mice and rats., Phytomedicine 2002 Mar; 9 (2): 135-41) [0005]
• - Lu Y and Westlund KN. (Effects of baclofen on colon inflammation-induced Fos, CGRP and SP expression in spinal cord and brainstem., Brain Res. 2001 Jan 19; 889 (1-2): 118-30) [0006]
• - Song DK, Suh HW, Huh SO, Jung JS, Him BM, Choi IG, and Kim YH. (Central GABAA and GABAB receptor modulation of basal and stress-induced plasma interleukin-6 levels in mice J Pharmacol Exp Ther. 1998 Oct; 287 (1): 144-9) [0007]
• - Taylor-Gjevre RM, and Gjevre YES. (Anti-glutamic acid decarboxylase antibodies in a patient with systemic lupus erythematosus and fibromyalgia symptoms., Lupus, 2005; 14 (6): 486-8) [0008]
• Bettler et al., Physiol. Rev. 2004, p. 857 [0030]
• - Bowery et al., Pharm. Rev. 2002, p. 254 [0030]

Claims (20)

Use of a GABA _B receptor selective agonist for the treatment of inflammatory skin diseases. Use according to claim 1, wherein the GABA _B receptor selective agonist is selected from the group of full agonists and partial agonists and the allosteric modulator group. Use according to claim 2, wherein the full agonist Baclofen or gabapentin is. Use according to claim 2, wherein the allosteric Modulator CGP 7930, GS 39783, CGP 44532 or CGP 13501 is. Use according to one of the preceding claims, wherein the inflammatory skin diseases are selected are from the group of erythematosquamous dermatoses, Autoimmune diseases, eczema, allergies, infectious diseases the skin and wound healing. Use according to claim 5, wherein the erythematosquamous Dermatosis psoriasis is. Use according to claim 5, wherein the autoimmune diseases blistering autoimmune dermatoses or collagenosis, in particular Lupus erythematosus, are. Use according to claim 5, wherein the eczema diseases Eczema or allergic contact dermatitis are. Use according to claim 5, wherein the infectious diseases the skin caused by viruses, bacteria, fungi or parasites become. Use according to any one of the preceding claims, wherein the GABA _B receptor selective agonist is administered to a patient having an inflammatory skin disease as defined in any of claims 5-9. Use according to claim 10, wherein the GABA _B receptor selective agonist, preferably baclofen, is administered at a dose of about 3 μg / kg body weight / day to about 5 mg / kg body weight / day. The use of claim 11, wherein the GABA _B receptor selective agonist, preferably baclofen, is administered at a dose of from about 10 μg / kg body weight / day to about 2.5 mg / kg body weight / day. Use according to any one of the preceding claims, wherein the GABA _B receptor selective agonist is administered in a pharmaceutical composition. Use according to one of the preceding claims, wherein the pharmaceutical composition is a pharmaceutically acceptable Carrier comprises. Use according to any one of claims 10-14, wherein administration of the GABA _B receptor selective agonist is local or systemic. Use according to claim 15, wherein administration of the GABA _B receptor selective agonist is oral, subcutaneous, intracutaneous, parenteral, transdermal, topical, intravenous, intraarterial, intramuscular, intraperitoneal, or in a combination of the foregoing ways. Use according to claim 16, wherein the administration transdermally or topically. Use according to claim 17, wherein the administration Transdermal takes place. Use according to claim 18, wherein the transdermal Administered in one dosage form selected from the group comprising spray, patch, ointment, semi-solid cream, Liquid, like a solution, solid, like a Powder, gel, emulsion or suspension. GABA _B receptor selective agonist as defined in any one of the preceding claims for the treatment of inflammatory skin diseases as defined in any one of the preceding claims.