WO2003097030A1

WO2003097030A1 - Use of 4-(4'-aminophenylsulphonyl)-benzoic acid and esters thereof as anti-inflammatory agents

Info

Publication number: WO2003097030A1
Application number: PCT/IB2003/002711
Authority: WO
Inventors: Michael D. Coleman; Sarah Hadley; Alan D. Perris; Karin Jorga; Joachim K. Seydel; Alexander Zolotoy
Original assignee: Immune Network Ltd
Current assignee: Immune Network Ltd
Priority date: 2002-05-21
Filing date: 2003-05-21
Publication date: 2003-11-27
Anticipated expiration: 2004-11-21
Also published as: AU2003238630A1

Abstract

Use of 4-(4'-aminophenylsulphonyl)- benzoic acid and esters thereof as antünflammatory agents is disclosed. The invention claims use of 4-(4'-aminophenylsulphonyl)- benzoic acid and esters thereof against diseases in which inflammation contributes to the pathology: Alzheimer's disease and other forms of dementia, asthma, dermatitis herpetiformis, AIDS pneumonia, Chronic Obstructive Pulmonary Disease, Amyotrophic Lateral Sclerosis, Parkinson's Disease, Multiple Sclerosis and others.

Description

• Use of 4-(4'-aminophenylsulphonyl)- benzoic acid and esters thereof, as anti- inflammatory agents

This application claims priority of Provisional Application Serial No. 60/382,394 filed May 21 , 2002, the disclosure of which is hereby incorporated by reference.

BACKGROUND OF THE INVENTION

1. Field of the invention

The present invention is directed to the use of derivatives of benzoic and anilinomethylcarboxylic acids and esters as anti-inflammatory agents.

2. Description of Related Art.

4,4'-diaminodiphenylsulphone (DDS) is used extensively against a variety of diseases such as leprosy, malaria, AIDS pneumonia, dermatitis herpetiformis, cutaneous manifestation of systemic lupus erythematosus (SLE) and many others (J. Uetrecht Clinics in Dermatology, 1989, 7, N3, 111-120) that are caused by plasmodia parasites, viruses, microbes and specific inflammatory processes. The use of DDS is significantly limited by two major adverse effects: hemolytic anemia (Jollow D. L. et al Drug Metab Rev. 1995;27(1 -2): 107-24) and ethemoglobinemia (Coleman M. D. Gen Pharmacol. 1995 26(7): 1461-7). Reduction of DDS's hemotoxicity while preserving its anti-microbial and anti-inflammatory properties is needed; with this in mind, a number of derivatives of DDS have been synthesized and evaluated for anti- inflammatory and anti-bacterial activity (Coleman M. D. et al J Pharm Pharmacol 1997, 49(1):53-7; Wiese M. et al. Arch Pharm (Weinheim). 1996, 329(3): 16 -8; J. K. Seydel et al. US Patent 5,084,449; Saxena M. et al Arzneimittelforschung, 1989 Sep;39(9):1081-4; Pieper H. et al. Arzneimittelforschung, 1989 Sep;39(9):1073-80.). However, most were anilino derivatives and showed only limited reductions in hemotoxicity while retaining strong anti-inflammatory properties. State of the art also indicated that derivatives of carboxylic acid such as indomethacin, diclofenac and aspirin as a rule do not show efficacy in those anti-inflammatory assays in which DDS is potent and effective.

These examples included: a) inhibition of eosinophil accumulation by DDS but not by aspirin and indomethacin

(Cook R. M Clin Exp Allergy. 1990, 20(5):511-7) • b) inhibition of myeloperoxidase activity by DDS, but not by indomethacin (Kettle A. J. Biochem Pharmacol. 1991 ,15; 41 (10): 1485-92)

c) inhibition of PAF-induced mortality by DDS but not by indomethacine or aspirin (Carlson R. P. et al Agents Action 1987, 21(3,4); 379-81)

d) inhibition of neuron death due to beta-Amyloid and microglia by DDS but not by indomethacin and ibuprofen (Giulian D. Patent WO 98/11923).

The inventors unexpectedly discovered that: the potency of derivatives of 4-(4'- aminophenyl-sulphonyl)- benzoic acid and esters thereof as anti-inflammatory agents is superior to that of DDS, while exhibiting significantly lower toxicity. For many such compounds, levels of methemoglobin are approximately twenty times less than observed with DDS.

Therefore, the present invention indicates a potential use of derivatives of 4-(4'- aminophenyl-sulphonyl)- benzoic acid and esters thereof in diseases in which inflammation contributes to the pathology such as Alzheimer's disease and other forms of dementia, AIDS pneumonia, dermatitis herpetiformis, asthma, Chronic Obstructive Pulmonary Disease, Amyotrophic Lateral Sclerosis, Parkinson's Disease, Multiple Sclerosis and others.

DETAILED DESCRIPTION OF THE INVENTION

Synthesis of 4-(4'-aminophenyl-sulphonyl)- benzoic acid and esters thereof (I) is described earlier (Jorga K 1990, Zum Wirkingsmodus und zur Antimycobacteriellen Wirkung von 4, (4'aminophenylsulphonyl)-benzoesaeureestern Ableiting: Quantitativer Structur-Wirkungs-Besiehungen. Dissertation, Universitaet Kiel). The invention is based on the discovery that compounds of formula (I) and pharmaceutically acceptable salts, complexes, chelates, hydrates, stereoisiomers, crystalline or amorphous forms, metabolites, metabolic precursors or prodrugs thereof demonstrate:

,a) potent anti-inflammatory activity - b) low levels of methemoglobin production

Thus, the present invention concerns the use of compounds of formula (I) and pharmaceutically acceptable salts, complexes, chelates, hydrates, stereoisomers, crystalline or amorphous forms, metabolites, metabolic precursors or prodrugs for the treatment of diseases with inflammation in which inflammation contributes to the pathology. The term "diseases in which inflammation contributes to the pathology" refers to: Alzheimer's disease and other forms of dementia, AIDS pneumonia, dermatitis herpetiformis, asthma, Chronic Obstructive Pulmonary Disease, Amyotrophic Lateral Sclerosis, Linear IgA bullous dermatosis, subcorneal pustular dermatosis, benign chronic bullous disease of childhood, SLE, cutaneous manifestation of SLE, pemphigus, pemphigoid, erythema elevatum diutinum, Sweet's syndrome, granuloma faciale, Henoch-Schonlein purpura, pioderma gangrenosum, hypocomplementemic urticarial vasculitus, discoid lupus, panniculitis, relapsing polychondritis, acne, alopecia mucinosa, pustular psoriasis, rheumatoid arthritis, brown ecluse spider bites, head injury, stroke, rheumatoid arthritis, chronic cholecystitis, bronciectasis, Hashimoto's thyroiditis, Crohn's disease, ulcerative colitis, silicosis, Parkinson's Disease, Multiple Sclerosis and others.

The compound of formula (I) is:

wherein, R is selected from:

R = H;

R = (CH₂)_nCH₃ (n=0, 1 , 2, 3,4, 5, 6, 7);

R = isopropyl, tert-butyl;

R = CH₂CH(CH₃)₂, CH₂C(CH₃)₃, CH₂CH₂CH(CH₃)_2> CH₂CH₂C(CH₃)₃; R = CH₂CH(CH₃)CH₂CH₃, CH₂C(CH₃)2CH₂CH₃;

R = cyclopentyl (C₅H₉), cyclohexyl (C_βHn), CH₂-C_εH₉, CHrCβHn;

-G^* wherein R1 is selected from

R = Cl, Br, F, CH₃, OH, OCH₃, OEt, CF₃, NH₂, NHCH₃, N(CH3)₂, CN, SO₂CH₃,

NHSO₂CH₃, NHS0₂Et, NCH₃SO₂CH₃, NCH₃SO₂Et, COCH₃, COEt, CONHCH3,

CONMeCHs, CONHEt, CONMe₂, CONMeEt.

The preferred formula includes R = H, CH_3l Bu and R = aryl with R₁ = 2-OH,

3-OH, 4-OH.

R2 = H, CH₃, Cl, F, OH, NH₂, OCH₃, CN, S0₂CH₃, N0₂.

Suitable routes of administration include parenteral injection, nasal administration, transdermal administration, rectal administration, inhalational administration, topical administration and oral administration.

Compounds of the present invention should be administered in association with one or more inert carriers, excipients and diluents. Assayable amounts of a compound of the invention will generally vary from about 0.001% to about 75% wt% of the entire weight of the composition. Preferred oral composition contains between 4% and about 50% of the active compound of formula (I). Preferred parenteral dosage includes between 0.01 to 10% by weight of active compound. Preferred topical • formulation contains a concentration of the compound of the invention from 0.1 to about 25% w/v (weight per unit volume).

Inert carriers include any material that does not degrade or otherwise covalently react with a compound of the invention.

Solid composition for oral administration may include: binders such as syrups, acacia, sorbitol, polyvinylpyrrolidone, carboxymethylcellulose, ethyl cellulose, microcrystalline cellulose or gelatin and mixtures thereof; excipients like starch, lactose or dextrins, disintegrating agents as alginic acid, sodium alginate, Primogel and the like; lubricants such as magnesium stearate, heavy molecular weight acids such as stearic acid, high molecular weight polymers such as polyethylene glycol; sweetening agents such as sucrose or saccharine; a flavoring agent such as peppermint, methyl salicylate or orange flavoring, and a coloring agent.

The liquid pharmaceutical compositions of the invention, whether they be solutions, suspensions or other like form may include: sterile diluents such as water for injection, saline solution preferably physiological saline, Ringer's solution, isotonic sodium chloride, fixed oils such as synthetic mono or diglycerides which may serve as the solvent or suspending medium, polyethylene glycols, glycerin, propylene glycol or other solvents. The compounds may be in the form of the free base or in the form of a pharmaceutically acceptable salt such as hydrochloride, sulfate, citrate, fumarate, methanesulfonate, acetate, tartrate, maleate, lactate, mandelate, salicylate and other salts known in the art.

EXAMPLE 1

Biological Materials and Methods-Methaemoglobin Studies

Sulphone-mediated methaemoglobin was generated in vitro using microsomal incubations in the presence of NADPH in single and two compartment systems. Rat (male Sprague-Dawley) microsomes were prepared as described previously and suspended in 50mM phosphate buffer (Purba et al., Br. J. Clin. Pharmacol. 1987, 23, 447), cytochrome P-450 levels were determined by the method of Omura and Sato (J. Biol. Chem. 1964, 239, 2370), to be 0.60 nmol/mg protein. All protein determination was carried out using 'Bio-Rad' (Bio-Rad Laboratories Ltd, Herts. UK; Bradford Anal. Biochem.1976, 72, 248) Microsomal suspensions (50mM phosphate buffer) of four human liver samples were used in these studies and were kindly provided by Drs. Barry Jones and Ruth Hyland, (Pfizer Central Research, Sandwich, UK) Livers 1-4 have been characterized using bufurolol (BUF), Testosterone (TEST) and diclofenac (DICL) assays to ascribe CYP 2D6, CYP 3A4 and 2C9 activities respectively. Human liver 1 (5.9 nmol P-450/ιmg protein) BUF metabolism was measured as 20.7 pmol/mg prot./min, while TEST metabolism was calculated to be 806 pmol/mg prot./min. DICL metabolism was 2620 pmol/mg prot./min. Liver 2 (2.29 nmol P-450/mg protein) BUF metabolism was measured as 23.7 pmol/mg prot./min, while TEST metabolism was calculated to be 540 pmol/mg prot./min. DICL metabolism was 1950 pmol/mg prot/min. Liver 3 (6.55 nmol P-450/mg protein) BUF metabolism was measured as 10.7 pmol/mg prot/min, while TEST metabolism was calculated to be 1739 pmol/mg prot/min. DICL metabolism was 1100 pmol/mg prot./min. Liver 4 (7.4 nmol P-450/mg protein) BUF metabolism was measured as 38.7 pmol/mg prot./min, while TEST metabolism was calculated to be 1129 pmol/mg prot./min: DICL metabolism was 2720 pmol/mg prot./min. The microsomes also contained cytochromes 2E1, 2C19, 2A6 and 1A2. The microsomal suspensions were stored at -70°C until utilised. Washed human erythrocytes were prepared in HEPES/Krebs-Henseleit buffer containing 10mM glucose (HEPES buffer) as previously described (Coleman and Jacobus Biochem. Pharmacol. 1993, 46,1363).

EXAMPLE 2 Biological Materials and Methods-Neutrophil Studies

Zymosan was opsonized by suspending 10 g zymosan in 1mL of guinea pig complement (Fife Technologies, Renfrew, Scotland) and incubating at 37°C for 20 min. The particles were then washed three times in phosphate buffered saline (PBS) and resuspended in RPMI (Sigma) and stored at 4°C until required. Whole blood was obtained by venepuncture from four healthy volunteers. Neutrophils in whole blood were challenged with yeast cell wall fragments (zymosan) which were coated with guinea pig complement to elicit a respiratory burst. Respiratory burst of the neutrophils was measured in whole blood by lucigenin-enhanced chemiluminescence based on the method of Tono-Oko et al. Clin. Immunol. Immunopathol.1983, 26, 66).

EXAMPLE 3 Experimental Design-Methaemoglobin studies

Two incubation systems were employed: the first (one compartment studies) was a mixture of 2 mg of rat liver microsomal protein suspension, NADPH (1mM final concentration), 500 L of washed erythrocytes plus the test compounds at a final concentration of 0.1 mM (added in 5 L DMSO; final incubation volume, 650 L). The incubations were carried out in quadruplicate and were placed in a gently shaking waterbath at 37°C for 45 mins. Samples were removed from the bath and stored on ice for not more than 5 min until methaemoglobin levels were measured on an lL-482 CO- oximeter (Instrumentation Laboratories, Warrington, UK). The amount of rat microsomal protein (2 mg) was employed in a previous study with the methaemoglobin former, DDS (Coleman et al., J. Pharm. Pharmacol.1991, 43, 779). The second incubation system involved the use of a two-compartment teflon system (Tingle et al. Br. J. Clin. Pharmacol. 1990, 30, 829) where compartment A contained the metabolising mixture (2mg rat microsomal protein, 1mM NADPH). The second compartment (B) containing the erythrocytes (500 L) was separated from A by a semi-permeable membrane (molecular weight cut off, 5000). In the single compartment studies, the compounds were added to a final concentration of 0.1 mM in 1mL volume. In the two compartment system, the compounds were added to a final concentration of 0.1 mM in compartment A, hence the compounds were diluted overall by 50% in the two compartment system and expected methaemoglobin levels would be half those of the single compartment system.

A second set of one-compartment stuJias was initiated, which involved the use of human liver microsomes from livers 1-4. Each incubation contained 3 mg of microsomal protein, 1mM final concentration of NADPH and each compound including DDS was tested at 100 M final concentration added in dimethyl sulphoxide, (DMSO; 1% final solvent concentration; final incubation volume, 650 L). All the microsomal incubations were incubated for 45 min at 37°C. Vehicle controls contained 1% DMSO. HEPES buffer and microsomal controls contained all the above except for NADPH. Methaemoglobin formation was determined as above and no more than background methaemoglobin was generated in NADPH-free incubations.

EXAMPLE 4

Experimental Design-Neutrophil studies.

Whole blood was incubated (15 min) with each of the 13 ester sulphone analogues or DDS prior to measurement of the respiratory burst. Briefly, 100 L of whole blood was added to 200 L of PBS and 100 L of lucigenin (1 X 10 /I) in a cuvette and equilibrated at 37°C with the compounds (final volume, 500 L). The concentrations for DDS and the ester analogues that were incubated with the mixture were 10, 20, 30, 40, 50 and 100 M. The pre-incubation allowed low background Relative Light Units (RLU) to be established following the trauma of isolation and mixing. Opsonized zymosan (100 L) was added to provide a stimulus to provoke a respiratory burst. Chemiluminescence readings in RLU at 425 nM were taken at 5 min intervals using a Bio-Orbit 1253 Luminometer (Labtech International, Sussex, UK). The samples were protected from light at all times. Generation of an oxidative burst in response to opsonized zymosan was used both as an index of cell viability and a positive control. IC-50 values for DDS and the 13 sulphone ester analogues were determined in comparison with the respiratory burst generated in the absence of the compounds. All statistical analysis was by one factor analysis of variance followed by Dunnett's test accepting P < 0.05 as significant. Log P estimates, which provide a guide to the lipophilicity of the compounds, were calculated using the ChemDraw Ultra package, version 6.0.1 (CambridgeSoft.com). All data is tabulated as mean + SD.

EXAMPLE 5 Methemoglobin and anti-inflammatory activity of some derivatives (I) versus DDS

% Methaemoglobin generated in 1 and 2 compartment systems by 13 sulphone esters (100 M per compound) in the presence of rat microsomes, NADPH and human erythrocytes, plus IC₅₀ values for the analogues for the suppression of opsonized zymosan-mediated neutrophil respiratory burst: both groups of values expressed as a percentage of the respective values generated by DDS under the same conditions. The lipophilicity of the compounds (Log P) was calculated using ChemDraw Ultra software. All values were significantly different from those of DDS (P < 0.001), except compounds 5 and 13 (see footnote) and Mean+/- SD, n = 4 per incubation.

. RB-Respiratory Burst % of DDS-mediated methaemoglobin

ICso % of DDS

Compound LOG P RB 1 Compartment 2 Compartment

1 2.5 45 3 300..22 ±+ 22..88 23.1+3.3

2 3.0 42 2 255..55 ±±11..00 14.0 + 3.3

3 2.6 61 2 2..22 ±+ 00..88 3.0 + 2.1

4 3.2 71 1 1..99 ++ 00..11 2.7 + 1.1

5 3.7 61 9 9..33 ±+ 33..33 90.5 + 15.6^'

6 4.1 87 4 4::11 ±+00..33 9.0 + 2.2

7 3.6 58 3 3..55 ±+ 00..33 4.8 + 0.1

8 3.6 38 1 100..44 ±+ 00..33 3.4 + 1.6

2.7 61 1 188..88 ++ 11..11 14.8±1.8

10 3.3 65 7 7..88 ±+ 00..88 6.4 + 2.0

11 4.1 65 4 4..88 ±+ 00..66 6.7+1.4

12 3.6 68 1 100..11 ±±11..77 16.6 ±3.1

13 1.9 116^* 5 5..22 ±+00..88 6.0 + 1.8

lues not significantly different from those of DDS

Compound R R2

1 CH2CH3 N

2 CH2CH2CH3 N

3 CH2CH3 H

4 CH2CH2CH₃ H

5 (CH2)3CH3 H

7 Cyclopentyl

8 Phenyl

9 Phenyl (2-OH) H

10 Phenyl (2-OCH₃) H

11 Phenyl (2-CH₃) H

12 CH2CH(CH3)2 H

13 H

Claims

We claim:

1. A method of treating diseases in which inflammation contributes to the pathology by administration to a patient in need thereof, of a pharmaceutically active amount of a compound having the following formula, or a pharmaceutically acceptable salt, complex, chelate, hydrate, stereoisomer, crystalline or amorphous form, metabolite, metabolic precursor, or prodrug thereof:

wherein, R is selected from the group consisting of H, (CH₂)_nCH₃ (n=0, 1, 2, 3,4, 5, 6, 7), isopropyl, tert-butyl, CH₂CH(CH₃)₂, CH₂C(CH₃)₃, CH₂CH₂CH(CH₃)₂, CH₂CH₂C(CH₃)₃ , CH₂CH(CH₃)CH₂CH₃, CH₂C(CH₃)₂CH₂CH3, cyclopentyl (C₅H₉), cyclohexyl (C₆Hn), CH₂-C₅H₉, CH₂-C₆Hn, and

where R1 is selected from the group consisting of Cl, Br, F, CH₃, OH, OCH₃, OEt, CF₃, NH₂, NHCH₃, N(CH3)₂, CN, SO₂CH₃, NHSO₂CH₃, NHSO₂Et, NCH₃S0₂CH₃, NCH₃S0₂Et, COCH₃, COEt, CONHCH3, CONMeCH₃, CONHEt, CONMe₂ and CONMeEt; and R2 is selected from the group consisting of H, CH₃, Cl, F, OH, NH₂, OCH3, CN, S0₂CH₃ and N0₂.

2. The method of claim 1 wherein R is selected from the group consisting of H, CH₃, and Bu, R is aryl and R-* is selected from the group conssisting of 2-OH, 3-OH or 4-OH.

3. The method of claim 1 wherein said disease is selected from the group consisting of Alzheimer's disease, dementia, asthma, AIDS pneumonia, dermatitis herpetiformis, Chronic Obstructive Pulmonary Disease, Amyotrophic Lateral Sclerosis, rheumatoid arthritis, linear IgA bullous dermatosis, subcorneal pustular dermatosis, benign chronic bullous disease of childhood, bullous eruptions of systemic lupus erythematosus, pemphigus, pemphigoid, erythema elevatum diutinum, Sweet's syndrome, granuloma faciale, Henoch-Schonlein purpura, pyoderma gangrenosum, hypocomplementic urticarial vasculitus, rheumatoid vasculitus, discoid lupus, systemic lupus erythematosus, cutaneous manifestation of systemic lupus erythematosus, panniculitis, relapsing polychondritis, acne, alopecia mucinosa, pustular psoriasis, brown recluse spider bites, Parkinson's Disease, Multiple Sclerosis, adverse effects caused by head trauma, adverse effects of hemorrhage caused by head trauma, AIDS Dementia Complex, encephalitis, and meningitis.

4. The method of claim 1 , wherein said disease is Alzheimer's disease.

5. The method of claim 1 , wherein said disease is dementia.

6. The method according to claim 1 , wherein said administration is a mode of administration selected from the group consisting of parenteral injection, nasal adminstration, transdermal administration, rectal administration, inhalational administration, topical administration and oral administration.

7. The method of claim 1, wherein said disease is a disease where inflammation and/or microbes contribute to the pathology.