WO1999045926A1

WO1999045926A1 - Condensed heterocyclic compounds as anti-inflammatory and immunomodulatory agents

Info

Publication number: WO1999045926A1
Application number: PCT/GB1999/000580
Authority: WO
Inventors: Patrick Vivian Richard Shannon; Thomas Eichholtz; David Linstead; Philip Masdin; Richard Skinner
Original assignee: University College Cardiff Consultants Limited
Priority date: 1998-02-27
Filing date: 1999-02-25
Publication date: 1999-09-16
Also published as: AU2632899A; WO1999045926B1; GB9804343D0

Abstract

The present invention relates to use of a compound of formula (1) as an immunomodulatory or anti-inflammatory drug or for use in the treatment of a therapeutic indication in which inhibition of dehydro-orate dehydrogenase (DHODH) is beneficial and salts and physiologically functional salts thereof, wherein X, R?5, R6, A, Z1 and Z2¿ have the meanings given in Claim 1.

Description

CONDENSED HETEROCYCLIC COMPOUNDS AS ANTI-INFLAMMATORY AND IMMUNOMODULATORY AGENTS

The present invention relates to the use of various heterocyclic compounds as immunomodulatory or anti-inflammatory drugs, or in the treatment of therapeutic indications relating to inhibition of dehydro-orate dehydrogenase. More specifically, the present invention relates to the use of Pyrrolo [3,2-b] carbazoles, Pyrrolo [2,3-b] carbazoles, 1H- Benzofuro [2,3-/] indoles, lH-Benzofuro [3,2- ] indoles, 1H-[1] Benzothieno [2,3- ] indoles, 1H-[1] Benzothieno [3,2-/] indoles, benzo[l,2-b:4,5- b']dipyrroles, benzo[2-b:5,4-b']dipyrroles, cyclopent[f]indoles, benzo[l,2-b:45- b']difurans, benzo(l,2-b:5,4-b']difurans, 2H-indeno[5,6-b]furans, benzo[l,2-b:4,5- b']dithiophenes, benzo[l,2-b:5,4-b']dithiophenes, cyclopent[f] indenes and 5H-furo[ 2,3-f]indoles for the aforementioned purposes.

The aforementioned heterocyclic compounds have been described in published International Patent Applications Nos WO94/02483, WO95/21170, WO95/21171 and WO96/01827, which also disclose anti tumour activity for the compounds.

Khoshtariya et al disclose the synthesis of certain indolobenzo[b] thiophenes and certain indolobenzo[b] furans, khim. Geterotsikl. Soedin (1980), (2) 203-8, and khim Geterotsiki Soedin (1984), (10)1366-70 respectively.

Kakhabrishvili et al, khim Geterotsikl Soedin (1985), (3) 355-8 disclose the synthesis of certain derivatives of indolo[5,6- ] and indolo [5,4-d] benzo[b] furans.

EP447,703 discloses the synthesis of certain benzo[5,6-b]benzofuran-2-carboxylates.

L.Chunchatprasert et al, J.Chem.Soc, Perkin Trans 1, 1779 (1992) disclose the synthesis of pyrrolo[3,2-b]carbazoles, lH-benzofuro[3,2-/]indoles and 1H-[1] benzothieno[2,3-

/] indoles.

Gruenhaus H., J. Heterocyclic Chem., 13(6) 1161-3 discloses the synthesis of certain . indenothiophenes.

It has now been found that the compounds employed in the present invention are inhibitors of dehydro-orate dehydrogenase (DHODH), and are useful as immunosuppressive and anti-inflammatory drugs and in the treatment of therapeutic indications in which inhibition of DHODH is beneficial.

According to the present invention there is provided use of a compound of formula (1) in the manufacture of a medicament for use as an immunomodulatory or anti- inflammatory drug or for use in the treatment of a therapeutic indication in which inhibition of dehydro-orate dehydrogenase (DHODH) is beneficial:-

and salts and physiologically functional salts thereof,

wherein X is O, S, SO, SO₂, CH₂, CO or NR⁷, wherein R⁷ is H or the following groups which may be optionally substituted: cyloalkyl, cycloalkenyl, alkyl, alkenyl, alkynyl, aryl, aralkyl, acyl, aroyl, sulphonyl, alkylsulphonyl, arylsulphonyl or COOR;

R⁵ and R⁶ are independently selected from H, hydroxy, nitro, amino, halo, cyano, CHO, COR , CO₂R and the following groups which may be optionally substituted: alkyl, aryl, aryloxy, aralkyloxy, alkoxy, aralkyl, wherein R⁸ is optionally substituted alkyl and aryl;

A is:

wherein R¹ is COR⁸, CHO, CH₂OH, CH₂OR⁸, CONH₂, COOR⁸, CONHR⁸, CONR⁸R⁸, CONHNR⁸R⁸, COO(CH₂)„NR⁸R⁸, CSOR⁸, CSSR⁸, COSR⁸, CSNHR⁸, CSNR⁸ R⁸, CNHOR⁸ or an optionally substituted 5 or 6 membered aromatic or nonaromatic heterocyclic ring containing 1 to 4 heteroatoms,

wherein the groups R⁸ are independently selected from hydrogen, optionally substituted alkyl, aryl, aralkyl, acyl, alkoxyalkyl, heterocycloalkyl and heteroaralkyl groups, and C _O optionally substituted hydrocarbyl groups which may contain one or two oxygen atoms in the chain and wherein n is 1 to 4;

or R may independently be sugar groups;

R² is H, hydroxy, haloalkyl, halo, cyano, COOR⁸, alkyl, aryl, alkenyl, alkynyl, alkoxy, (wherein alkyl, aryl, alkenyl, alkynyl and alkoxy can be substituted) CH₂CH₂CO₂R⁹ (wherein R⁹ is alkyl or aryl), CHO, COR⁸, COOR⁸ or a C_H0 optionally substituted hydrocarbyl group which may contain one or two oxygen atoms in the chain, wherein R⁸ is independently selected from the groups defined for R⁸ above;

Y is O, S, SO, SO₂, CH₂, CO or NR⁷ wherein R⁷ is independently selected from groups hereinbefore defined for R ;

Z¹ and Z² are independently selected from H, halogen, cyano, amino, alkyl, COOR⁸, CONHR⁸, COR⁸, CH₂OH, CH₂OR⁸, CONH₂, CON R⁸R⁸, CSOR⁸, CSSR⁸, COSR⁸, CSNHR⁸, CSNR⁸R⁸ and CNHOR⁸ wherein R⁸ is independently selected from the groups defined for R above; or Z and Z together form the group:

wherein R³ and R⁴ are independently selected from H, hydroxy, alkyl, haloalkyl, alkoxy, halo, cyano, azido, nitro, amino, alkyl amino, dialkyl amino, CHO, COR⁸, CONHR⁸, CON R⁸R⁸ (wherein R⁸ is independently selected from the groups defined for R⁸ above ),carboxyl or CO₂R¹⁰, wherein R¹⁰ is independently selected from alkyl, aralkyl and aryl. The term hydrocarbyl includes strajght-chain or branched alkyl, alkenyl and alkynyl groups; cycloalkyl, cycloalkenyl and cycloalkynyl groups; and aralkyl, aralkenyl and aralkynyl groups where the alkyl, alkenyl or alkynyl portion may be straight-chain or branched.

Alkyl groups may be straight or branched chain alkyl groups, and may contain 1-10 carbon atoms and suitably 1-6 carbon atoms. Examples of such alkyl groups include methyl, ethyl, t-butyl and the like.

Alkenyl groups may be straight or branched chain alkenyl groups, and may contain 2-10 carbon atoms and suitably 2-6 carbon atoms. Examples of such alkenyl groups include ethenyl, butenyl and the like.

Alkynyl groups may be straight or branched chain alkynyl groups, and may contain 2- 10 carbon atoms and suitably 2-6 carbon atoms. Examples of such alkynyl groups include ethynyl, propynyl and the like.

Haloalkyl groups may be straight or branched chain haloalkyl groups and may contain 1-10 carbon atoms and suitably 1-6 carbon atoms. Such groups may contain one or more halo atoms. Examples of haloalkyl groups include trifiuoromethyl, and the like.

Acyl groups are derived from carboxylic acids and may be straight or branched and may contain 1-10 carbon atoms and suitably 1-6 carbon atoms. Examples of suitable acyl groups include ethanoyl and propanoyl groups.

Alkoxy may be straight or branched and may contain 1-10 carbon atoms and suitably 1- 6 carbon atoms. Examples of suitable alkoxy groups include methoxy, ethoxy and the like.

Aryl includes both carbocychc aryl groups and heterocyclic aryl groups normally containing a maximum of 10 ring atoms. Carbocychc aryl groups include, eg phenyl and naphthyl and contain at least one aromatic ring. Heterocyclic aryl groups include eg thienyl, Riryl, pyridyl, indolyl and quinolinyl rings.

An aralkyl group may contain from 1 to 4 atoms in the alkyl portion and the aryl portion may be a carbocychc or heterocyclic aryl group.

A Ci-io hydrocarbyl group optionally containing one or two oxygen atoms includes alkyl, hydroxyalkyl, alkenyl, alkynyl, Ci-io carbamoylalkyl, Cι-₁₀ alkoxyalkyl, cycloalkyl, cycloalkenyl, aralkyl, C_Mo aryloxyalkyl, acyl or aryl.

Hydrocarbyl, aryl, alkyl, alkenyl, alkynyl and aralkyl groups may be optionally substituted by hydroxy, azido, alkenyl, halo, nitro, amino (optionally substituted by one or 2 alkyl groups), cyano, carboxylate, alkyl ester, aralkyl ester, aryl ester (wherein the alkyl ester, aralkyl ester and aryl ester can be substituted) alkyl, aryl, aralkyl, aryloxy, arylalkoxy, substituted arylalkoxy, sulphinyl, sulphonyl, thio, Cπo alkylthio, alkoxy, hydroxyalkyl, haloalkyl, phosphate, phosphonate, silyl, silyloxy, (wherein silyl and silyloxy may be substituted by one or more Cι_-6 alkyl or aryl groups) keto or, formyl.

Cycloalkyl includes both cycloalkyl groups and heterocycloalkyl groups normally containing between 3 and 6 ring atoms. Heterocycloalkyl groups include e.g. morpholino, thiomorpholino, piperidino, imidazolino, imidazolidino, pyrrolidino, pyrazohdino, piperazino, tetrahydrofuranyl, tetrahydropyranyl. Cycloalkyl groups include C₃-₆ carbocycles such as cyclopentyl and cyclohexyl.

Cycloalkenyl includes both cycloalkenyl groups and heterocycloalkenyl groups normally containing between 3 and 6 ring atoms.

Substituents which may be present on alkyl esters, aralkyl esters and aryl esters include nitro, amino, hydroxy, alkoxy, halogen, cyano or alkyl.

Examples of suitable aromatic 5- or 6-membered rings containing 1 to 4 heteroatoms, include oxadiazole, oxazole, isoxazole, imidazole, pyrazole, triazole, tetrazole, pyrimidine, pyrazine, pyridazine, triazine, thiadiazole, thiazole, isothiazole.

Examples of suitable non-aromatic 5- or 6- membered rings containing 1 to 4 heteroatoms, include oxazoline, oxazolidine, thiazoline, thiazolidine, oxazolidinone, thiazolidinone, imidazoline, imidazolidine, pyrazolidine and pyrazoline.

Substituents which may be present on R¹ include azido, nitro, cyano, halo, haloalkyl, hydroxy, CHO, COR⁸ , CO₂R⁸, CONHR⁸, CONR⁸R⁸, oxo or the following groups which may be optionally substituted: alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, aryl, aralkyl, heteroaryl, acyl, aroyl, aralkoyl, alkoxy or amino.

Substituents which may be present on cycloalkyl, cycloalkenyl, alkyl, alkenyl, alkynyl, alkoxy, aryl, aralkyl, acyl, aroyl, alkylsulphonyl or arylsulphonyl groups include alkyl, alkoxy, halo, sulphinyl, hydroxy, amino (optionally substituted by one or two alkyl groups or part of a heterocyclic ring), haloalkyl (eg trifluoromethyl), sulphonyl, cyano, nitro or azido.

Substituents which may be present on the sulphonyl and sulphinyl include alkyl, aryl and aralkyl.

Halo represents fluoro, chloro, bromo or iodo.

Where R is a sugar this group may be present in a protected or unprotected form. Preferred sugar-protecting groups include isopropylidene, benzylidene acetate, benzoyl, paranitrobenzyl, paranitrobenzoyl, benzyl, substituted silyl and tetrahydropyranyl.

When R is a sugar such as a tetrose, pentose, hexose (including furanose and pyranose) or heptose, preferred sugars include glucose, fructose, mannose, ribose, arabinose.

X preferably represents S, O or NH; more preferably S or NH; more preferably NH.

R⁵ and R⁶ are preferably independently selected from H, alkyl and aryl; more preferably from H and alkyl.

Y preferably represents NH.

In a first series of preferred compounds there is provided a compound of the general formula (II)

and salts and physiologically functional derivatives thereof,

wherein A is as hereinbefore defined, X is O, S, SO, SO₂, CH₂, CO or NR⁷, wherein R⁷ is H, alkyl, aralkyl, aryl, alkenyl, acyl, alkynyl, sulphonyl or substituted sulphonyl;

Y is O, S, SO, SO₂, CH₂, CO or NR⁷;

R¹ is COR¹¹, COOR¹¹, CHO, CH₂OH, CH₂OR¹², CONH₂, CONHR¹³R¹⁴, CONHR¹³, CONR¹³R¹⁴, COO(CH₂)_nNR¹³R¹⁴, wherein R¹¹ is H, alkyl, aryl, substituted aryl or aralkyl, R¹² is acyl or substituted acyl, R¹³ and R¹⁴ are independently hydrogen, alkyl or aryl, and n is 1 to 4 carbon atoms;

R² is H, COOR¹¹, alkyl, aryl, substituted aryl or CH₂CH₂CO₂R⁹ wherein R⁹ is alkyl or aryl;

R and R⁴ are independently H,hydroxy, alkyl, haloalkyl, alkoxy, halo, cyano, nitro, amino, alkyl amino, dialkyl amino, substituted alkyl, carboxyl or CO R⁹;

R⁵ is H, alkyl, substituted alkyl, aralkyl, nitro, amino, halo, cyano, CHO, COOR¹¹;

R⁶ is H, aryl, alkyl, aralkyl, nitro, halogen, CHO or COR¹⁵ wherein R¹⁵ is alkyl or aryl.

In the first series of preferred compounds, it is further preferred tha -

X is O, S, SO, SO₂, CH₂₎ CO or NR⁷ , wherein R⁷ is H, alkyl, aralkyl, aryl, alkenyl, acyl, alkynyl or sulphonyl;

Y is O, S, SO, SO₂,CH₂, CO orNR⁷ ;

R¹ is COORⁿ,CHO,CH₂OH, CH₂OR¹², CONH₂ , CONHR¹³ or CONR¹³R¹⁴, wherein R is H, alkyl, aryl, substituted aryl or aralkyl, R is acyl or substituted acyl, and R and R are independently alkyl or aryl;

R³ and R⁴ are independently H, hydroxy, alkyl, haloalkyl, alkoxy, halo, cyano, nitro, amino, alkyl amino, dialkyl amino, substituted alkyl, carboxyl or CO₂R⁹;

R⁵ is H, alkyl, substituted alkyl, aralkyl, nitro, halo, cyano, CHO;

R⁶ is H, alkyl, aralkyl, nitro, halo, CHO or COR¹⁵ wherein R¹⁵ is alkyl or aryl;

X is preferably O, S or NR⁷, wherein R⁷ is H, alkyl, sulphonyl or toluene sulphonyl;

Y is preferably NR⁷;

R¹ is preferably COR¹¹ , COOR¹¹ , CH₂OR¹², CONH₂, CNHNR¹³R¹⁴, CONHR¹³, CONR¹³ R¹⁴, COO(CH₂)_n NR¹³R¹⁴, wherein R¹¹ is H, alkyl, aryl, substituted aryl or aralkyl, R¹² is acyl or substituted acyl, and R¹³and R¹⁴ are independently hydrogen, alkyl or aryl and n is 1 to 4 carbon atoms;

R² is preferably COOR¹¹, alkyl or CH₂CH₂CO₂R⁹ wherein R⁹ is alkyl or aryl;

R³ and R⁴ represent independently H,hydroxy, alkyl, alkoxy, halogen, cyano, substituted alkyl or carboxyl;

R⁵ is preferably H or alkyl;

R⁶ is preferably H, alkyl or aryl and salts.

X preferably represents S or NH, A is preferably

and Y preferably represents NH.

R¹ is preferably COOR¹¹ , with R¹¹ preferably being alkyl or aralkyl.

R² is preferably H or alkyl. R³ is preferably H, alkoxy or halo.

R⁴ is preferably H, alkoxy or halo.

R⁵ is preferably alkyl and

R⁶ is preferably H

Particularly preferred compounds include:

3-Pyridyl 3, 4-dimethylpyrrolo( 3, 2-b] carbazole-2-carboxylate

[(3-Dimethylamino)phenyl]3, 4-dimethylpyrrolo[3,2-b] carbazole-2-carboxylate

Benzyl 1, 3, 4- trimethylpyrrolo( 3, 2-b] carbazole-2-carboxylate

Phenyl 3, 4-dimethylpyrrolo [ 3, 2-b] carbazole-2-carboxylate 3,4-Dimethyl-2-( 1-imidazolylcarbonyl ) pyrrolo [3, 2-b] carbazole

Ethyl 3, 4-dimethylpyrrolo [3,2,-b]carbazole-2-carboxylate;

Ethyl 3, 4-dimethylbenzothieno [4, 5-f] indole-2-carboxylate;

Benzyl 3, 4-dimethylpyrrolo [3, 2-b] carbazole-2-carboxylate;

Benzyl 8-f luoro-3, 4-dimethylpyrrolo [3,2-b] carbazole-2-carboxylate; Ethyl 8-f luoro-3, 4-dimethylpyrrolo [3,2-b] carbazole-2-carboxylate

Benzyl 3,4, 6-trimethylpyrrolo (3, 2-b] carbazole-2-carboxylate;

Ethyl 3,4, 6-trimethylpyrrolo [3, 2-b] carbazole-2-carboxylate;

8-Fluoro-3, 4-dimethylpyrrolo [ 3, 2-b] carbazole-2-carboxylic acid

3,4-Dimethylpyrrolo[ 3, 2-_b] carbazole-2-carboxylic acid; Ethyl 8-methoxy-3,4-dimethylpyrrolo [3, 2-b] carbazole-2-carboxylate;

3,4,6-Trimethylpyrrolo [3, 2-b] carbazole-2-carboxylic acid and

Benzyl 8-methoxy-3, 4-dimethylpyrrolo [3, 2-Jb] carbazole-2-carboxylate;

and physiologically functional derivatives thereof.

Especially preferred is Ethyl 3, 4-dimethylpyrrolo [3,2,-b]carbazole-2-carboxylate and physiologically functional derivatives thereof.

Compounds in the first series may be prepared according to the reaction schemes and procedures described in published International Patent Application No. WO94/02483, incorporated herein by reference.

A second series of preferred compounds have the formula (III)

and salts and physiologically functional derivatives thereof, wherein A is as hereinbefore defined,

X is O, S, SO, SO₂, CH₂, CO or NR⁷, wherein R⁷ is H, alkyl, aralkyl, aryl, alkenyl, acyl, alkynyl, sulphonyl or substituted sulphonyl;

Y is O, S, SO, SO₂, CH₂, CO or NR⁷;

R¹ is COOR¹⁶, CONHR¹⁶, CONR¹⁶R¹⁷, CSOR¹⁶, CSSR¹⁶, COSR¹⁶, CSNHR¹⁶, CSNR¹⁶R^π CNHOR¹⁶ wherein R¹⁶ and R¹⁷ are independently C_1-10 optionally substituted hydrocarbyl groups which may optionally contain one or two oxygen atoms in the chain; or R¹⁶ and R¹⁷ are independently alkoxyalkyl, heterocycloalkyl, heteroaralkyl,

or R¹⁶ and R¹⁷ may independently be sugar groups;

1 (

R is H, halo, cyano, COOR , alkyl, aryl, alkenyl, alkynyl, alkoxy, (wherein alkyl, aryl, alkenyl, alkynyl and alkoxy can be substituted) or CH₂CH CO₂R⁹ wherein R⁹ is alkyl or aryl;

R³ and R⁴ are independently H,hydroxy, alkyl, haloalkyl, alkoxy, halo, cyano, nitro, amino, alkyl amino, dialkyl amino, substituted alkyl, carboxyl or CO₂R ;

R⁵ is H, hydroxy, aryloxy, aralkyloxy, alkyl, substituted alkyl, aralkyl, nitro, amino, halo, cyano, CHO; and

Particularly preferred compounds in the second series have the formula (IV)

wherein,

X preferably represents S,O or NH;

R¹ is preferably COOR¹⁶, with R¹⁶ preferably being a group of formula

-(CH₂COO)„ Z

where n is 0 or 1 and

Z is a phenyl or benzyl group optionally substituted by one or more groups selected from hydroxy, carboxyl, nitro, amino, phthalimido, p_-nitrobenzyl and p_- nitrobenzyloxy;

or Z is a C_1- straight or branched alkyl or cycloalkyl group optionally substituted by one or more groups selected from hydroxy, carboxyl, halo, amino, dialkylamino, alkylsulphinyl, alkylsulphonyl and benzyloxy;

or Z is a substituted glucofuranosyl moiety;

R² is preferably H or alkyl; R³ is preferably H, alkoxy, hydroxy or halo;

R⁴ is preferably H, alkoxy, hydroxy or halo;

R⁵ is preferably alkyl; and

R⁶ is preferably H.

and salts and physiologically functional derivatives thereof.

Particularly preferred compounds include:

[(2-Dimethylamino)ethyl] 3,4-dimethylpyrrolo[3,2-b]carbazole-2-carboxylate (2-Methylsulphonylethyl) 3,4-dimethylpyrrolo[3,2-b]carbazole-2-carboxylate (2-Methylsulphinylethyl) 3,4-dimethylpyrrolo[3,2-b]carbazole-2-carboxylate (l,3-Dibenzyloxypropyl-2) 3,4-dimethylpyrrolo[3,2-b]carbazole-2-carboxylate (l-Benzyloxy-3-hydroxypropyl-2) 3,4-dimethylpyrrolo[3,2-b]carbazole-2-carboxylate (l,3-Dihydroxypropyl-2) 3,4-dimethyl-pyrrolo[3,2-b]carbazole-2-carboxylate (2-Amino-2-methylpropyl-l) 3,4-dimethylpyrrolo[3,2-b]carbazole-2-carboxylate (4-Nitrophenylmethyl) 2-(3,4-dimethylpyπOlo[3,2-b]carbazole-2-carboxy)acetate 2-(3,4-Dimethylpyrrolo[3,2-b]carbazole-2-carboxy)acetic acid Cyclohexyl 3,4-dimethylpyrrolo[3,2-b]carbazole-2-carboxylate Cyclohexylmethyl 3,4-dimethylpyrrolo[3,2-b]carbazole-2-carboxylate Cyclopentyl 3,4-dimethylpyrrolo[3,2-b]carbazole-2-carboxylate Cyclooctyl 3,4-dimethylpyrrolo[3,2-b]carbazole-2-carboxylate

3,5-Di(tert-butyldiphenylsilyloxy)cyclohexyl 3,4-dimethylpyrrolo[3,2-b]carbazole-2- carboxylate

3,5-Dihydroxycyclohexyl 3,4-dimethylpyrrolo[3,2-b]carbazole-2-carboxylate cis-4-tert-Butyldiphenylsilyloxycyclohexyl 3,4-dimethylpyrrolo[3,2-b]carbazole-2- carboxylate cis-4-Hydroxycyclohexyl 3,4-dimethylpyπOlo[3,2-b]carbazole-2-carboxylate trans-4-tert-Butyldiphenylsilyloxycyclohexyl 3,4-dimethylpyrrolo[3,2-b]carbazole-2- carboxylate trans4-Hydroxycyclohexyl 3,4-dimethylpyrrolo[3,2-b]carbazole-2-carboxylate Tetrahydro-2H-pyran4-yl 3,4-dimethylpyrrolo[3,2-b]carbazole-2-carboxylate l-Benzylpiperidin-4-yl 3,4-dimethylpyrrolo[3,2-b]carbazole-2-carboxylate Piperidin-4-yl 3,4-dimethylpyrrolo[3,2-b]carbazole-2-carboxylate l-Methylpiperidin-4-yl 3,4-dimethylpyπOlo[3,2-b]carbazole-2-carboxylate (3,4-Dimethylpyrrolo[3,2-b]2-carbazolyl)3-O-( l,2:5,6-di-O-isopropylidene glucofuranoside)

(3,4-Dimethylpyrrolo[3,2-b]2-carbazolyl) 3-O-(l,2-O-isopropyl-ideneglucofuranoside)

[3-(4-Nitrophenylmethoxy)phenyl] 3,4-dimethylpyrrolo[3,2-b]carbazole-2-carboxylate

(3-Hydroxyphenyl) 3-4,-dimethylpyrrolo[3,2-b]carbazole-2-carboxylate (4-Phthalamidophenyl) 3,4-dimethylpyrrolo[3,2-b]carbazole-2-carboxylate

4-(Aminophenyl) 3,4-dimethylpyrrolo[3,2-b]carbazole-2-carboxylate

(4-Nitrophenylmethyl) 3-(3,4-dimethylpyrrolo[3,2-b]carbazole-2-carboxy)benzoate

3-(3,4-Dimethylpyrrolo[3,2-b]carbazole-2-carboxy)benzoic acid 3-(tert-Butyldiphenylsilyloxymethyl)phenyl 3,4-dimethylpyrrolo[3,2-b]carbazole-2- carboxylate

(3-Hydroxymethyl)phenyl 3 ,4-dimethylpyrrolo[3,2-b]carbazole-2-carboxylate

[3-(4-Nitrophenylmethoxy)phenyl] 4-methyl-lH-[l ]benzothieno[2,3 -f]indole-2- carboxylate

(3-Hydroxyphenyl)4-methyl lH-[l]benzothieno[2,3-f]indole-2-carboxylate 3-(4-Nitrophenylmethoxy)phenylmethyl 3,4-dimethylpyrrolo[3,2-b]carbazole-2- carboxylate

3-(tert-Butyldiphenylsilyloxyphenyl)methyl 3,4-dimethylpyrrolo[3,2-b]carbazole-2- carboxylate (3-Hydroxyphenyl)methyl 3 ,4-dimethylpyrrolo[3,2-b]carbazole-2-carboxylate (1 -Hydroxy-3-methylpropyl-2) 3,4-dimethylpyrrolo[3,2-b]carbazole-2-carboxamide 2-Hydroxyethyl 3,4-dimethylpyrrolo[3,2-b]carbazole-2-carboxamide 2-Chloroethyl 3,4-dimethylpyrrolo[3,2-b]carbazole-2-carboxamide N-(2-Aminoethyl) 3,4-dimethylpyrrolo[3,2-b]carbazole-2-carboxamide N-(2-Acetamidoethyl) 3,4-dimethylpyrrolo[3,2-b]carbazole-2-carboxamide (3-Aminopropyl- 1) 3,4-dimethylpyrrolo[3,2-b]carbazole-2-carboxamide 2-Hydroxyethyl 4-methyl- lH-[l]benzothieno[2,3-f]indole-2-carboxamide 2-Chloroethyl 4-methyl-lH-[l]benzothieno[2,3-f]indole-2-carboxamide

and physiologially functional derivatives thereof.

Compounds in the second series may be prepared according to the reaction schemes and procedures described in published International Patent Application No. WO95/21170, incorporated herein by reference.

In a third series of preferred compounds there is provided a compound of the general formula (V)

or a salt or physiologically functional derivative thereof, wherein A is as hereinbefore defined

X is O, S, SO, SO₂, CH₂, CO or NR⁷, wherein R⁷ is H, alkyl, aralkyl, aryl, alkenyl, acyl, alkynyl, sulphonyl, substituted sulphonyl, or COOMe;

Y is O, S, SO, SO₂, CH₂, CO or NR⁷;

R¹ is COR¹⁸, CHO, CH₂OH, CH₂OR¹⁸, CONH₂, COOR¹⁸, CONHR¹⁸, CONR¹⁸R¹⁹, CSOR¹⁸, CSSR¹⁸, COSR¹⁸, CSNHR¹⁸, CSNR¹⁸R¹⁹, CNHOR¹⁸ wherein R¹⁸ and R¹⁹ are independently hydrogen, alkoxyalkyl, heterocycloalkyl, heteroaralkyl, or C_1-10 optionally substituted hydrocarbyl group which may optionally contain one or two oxygen atoms in the chain;

or R¹⁸ and R¹⁹ may independently be sugar groups;

R² is H, halo, cyano, COOR¹⁸, alkyl, aryl, alkenyl, alkynyl, alkoxy, (wherein alkyl, aryl, alkenyl, alkynyl and alkoxy can be substituted) or CH₂CH₂CO₂R⁹ wherein R⁹ is alkyl or aryl;

Z¹ is H, alkyl, halogen, cyano, amino, COOR , 1¹8⁶, CONHR¹⁸, COR¹⁸, CH₂OH, CH2OR¹⁸, CONH₂, CONR¹⁸R¹⁹, CSOR¹⁸, CSSR¹⁸, COSR¹⁸, CSNHR¹⁸, CSNR¹⁸R¹⁹ or CNHOR¹⁸; Z² is H, halogen, cyano, amino, alkyl, COOR¹⁸, CONHR¹⁸, COR¹⁸, CH₂OH, CH₂OR¹⁸, CONH₂, CONR¹⁸R¹⁹, CSOR¹⁸, CSSR¹⁸, COSR¹⁸, CSNHR¹⁸, CSNR¹⁸R¹⁹ or CNHOR¹⁸;

R⁵ is H, hydroxy, aryloxy, aralkyloxy, alkyl, substituted alkyl, aralkyl, nitro, amino, halo, cyano, COOR¹⁸ or CHO;

R⁶ is H, aryl, alkyl, aralkyl, nitro, halogen, CHO or COR²⁰ wherein R²⁰ is alkyl or aryl.

X preferably represents NH, A is preferably

and Y preferably represents NH.

R¹ is preferably COOR¹⁸, with R¹⁸ preferably being alkyl or aralkyl.

R is preferably H, alkyl, or COOR wherein R is preferably alkyl,

Z¹ is preferably alkyl

Z² is preferably alkyl or COOR¹⁸.

R⁵ is preferably hydrogen and

R⁶ is preferably hydrogen or methyl.

Preferred groups of compounds include:

Ethyl 1 ,7-dihydro-3,4,6-trimethylpyrrolo[3,2-fjindole-2-carboxylate; Diethyl l,7-dihydro-3,4,6-trimethylpyrrolo[3,2-f] indole-2,5-dicarboxylate; and Ethyl 6-methoxycarbonyl-3 , 4-dimethylpyrrolo [3 ,2-f]indole-2-carboxylate

and physiologically functional derivatives thereof; and

Ethyl 6-Benzyloxycarbonyl-3, 4-dimethylpyrrolo [3, 2-f ]indole-2-carboxylate; Dibenzyl 3,4-dimethylpyrrolo[3,2-f]indole-2,6-dicarboxylate; Ethyl 7-methoxycarbonyl-3,4-dimethylpyrrolo [3,2-f]indole-2-carboxylate; and Ethyl 3,4-dimethylpyrrolo[3,2-f]indole-2-carboxylate and physiologically functional derivatives thereof.

Compounds in this third series may be prepared according to the reaction schemes and procedures described in published International Patent Application No. WO95/21171.

In a fourth series of preferred compounds there is provided a compound of the general formula (VI)

and salts and physiologically functional derivative thereof, wherein B is

X is O, S, SO, SO₂, CH₂, CO or NR⁷, wherein R⁷ is H or the following groups which may be optionally substituted: cyloalkyl, cycloalkenyl, alkyl, alkenyl, alkynyl, aryl, aralkyl, acyl, aroyl, alkylsulphonyl or arylsulphonyl;

Y is O, S, SO, SO₂, CH₂, CO or NR⁷; R¹ is an optionally substituted 5- or 6-membered heterocyclic ring containing 1 to 4 heteroatoms wherein the 5- or 6- membered ring may be aromatic or non-aromatic;

R² is H, hydroxy, halo, haloalkyl, cyano, alkyl, aryl, alkenyl, alkynyl, alkoxy, (wherein alkyl, aryl, alkenyl, alkynyl, and alkoxy can be substituted), CHO, COR²³, COOR²³ wherein R²³ is hydrogen or is a C_MO optionally substituted hydrocarbyl group which may contain one or two oxygen atoms;

R³ and R⁴ are independently H, hydroxy, alkyl, haloalkyl, azido, CHO, COR²³, CO₂R²³, CONHR²³, CONR²³R²⁴, alkoxy, halo, cyano, nitro, amino, alkyl amino, dialkyl amino, carboxyl wherein R²⁴ is alkyl, aryl or aralkyl;

R²¹ is H, hydroxy, nitro, amino, halo, cyano, CHO, COR²³, or the following groups which may be optionally substituted: alkyl, aryl, aryloxy, aralkyloxy, alkoxy, aralkyl;

R²² is H, hydroxy, amino, nitro, halo, CHO, COR²⁵, CO₂R²⁵ wherein R²⁵ is optionally substituted alkyl or aryl, or R⁶ is alkyl, aralkyl, or aryl wherein alkyl, aralkyl or aryl may be optionally substituted.

Suitably X is O,S, SO, SO₂,CH₂,CO or NR⁷ wherein R⁷ is suitably H, alkyl, aralkyl, aryl, alkenyl, acyl, alkynyl or optionally substituted sulphonyl;

Suitably R¹ is an optionally substituted five or six-membered heterocyclic ring containing one or two nitrogen atoms and optionally one other heteroatom.

Suitably R² is H, alkyl or COOR²³ wherein R²³ is as defined above;

Suitably R³ and R⁴ are independently H, hydroxy, alkyl, haloalkyl, alkoxy, halo, cyano, nitro, amino, alkylamino, dialkylamino or substituted alkyl;

Suitably R²¹ is H, alkyl, substituted alkyl, aryl, aralkyl, nitro, halo, cyano or CHO;

Suitably R²² is H, alkyl, aralkyl, nitro, halo, CHO or COR²⁵wherein R²⁵ is suitably alkyl or aryl.

X preferably represents S or NH, B is preferably

and Y preferably represents NH.

R¹ is preferably an optionally subsituted five-membered ring containing two nitrogen atoms and one oxygen atom wherein the 5-membered ring may be aromatic or nonaromatic. Preferred substituents are alkyl, aryl or aralkyl;

R is preferably H or C alkyl;

R is preferably H, alkoxy, halo or hydroxy;

R is preferably H or alkyl; and

R is preferably H or alkyl.

Particularly preferred compounds include:

3,4-Dimethyl-2-(3-ethyl-l,2,4-oxadiazol-5-yl) pyrrolo[3,2-b]carbazole;

2-(3-Benzyl- l,2,4-oxadiazol-5-yl)-3,4-dimethylpyrrolo[3,2-b]carbazole;

3,4-Dimethyl-2-(3-ethyl- 1 ,2,4-oxadiazol-5-yl) pyrrolo[2,3-b]carbazole; 2-(3-ethyl-l,2,4-oxadiazol-5-yl)-4-methyl-lH-[l]benzothieno[2,3-/]indole;

3,4-Dimethyl-2-(2-methyl-l,3,4-oxadiazol-5-yl)-pyrrolo[3,2-b]carbazole;

3,4-Dimethyl-2-(2-ethyl-l,3,4-oxadiazol-5-yl)-pyrrolo[3,2-/3]carbazole;

3,4-Dimethyl-2-(2-phenyl-l,3,4-oxadiazol-5-yl)-pyrrolo[3,2-b]carbazole;

2-(2-Ethyl-l,3,4-oxadiazol-5-yl)4-methyl-lH-[l]benzothieno[2,3-/]indole; 3,4-Dimethyl-2-[3-(3,4-methylenedioxyphenyl)-l,2,4-oxadiazol-5-yl]pyrrolo[3,2- b]carbazole;

4-Methyl-2-(2-oxazolin-2-yl)- lH-[ 1 ]benzothieno[2,3-/]indole

3,4-Dimethyl-2-(3-methyl-l,2,4-oxadiazol-5-yl)pyrrolo[3,2-/>]carbazole 3,4-Dimethyl-2- 3 -methyl- 1 ,2 ,4-oxadiazol-5 -yl)pyrrolo [2,3-/3] carbazole 3,4-Dimethyl-2- (3-phenyl)-l,2,4-oxadiazol-5-yl]pyrrolo[3,2-b]carbazole 3,4-Dimethyl-2- 2-oxazolin-2-yl)pyrrolo[3,2-b]carbazole 3,4-Dimethyl-2- 3-(l-piρeridinylmethyl)-l,2,4-oxadiazol-5-yl)pyrrolo[3,2- b]carbazole 3,4-Dimethyl-2- 3-(4-pyridyl)-l,2,4-oxadiazol-5-yl]pyrrolo[3,2-δ]carbazole 3,4-Dimethyl-2- 3 -methoxymethyl- 1 ,2,4-oxadi azol- 5 -yl)pyrrolo [3 ,2-b] carbazole 3,4-Dimethyl-2- 3-(2-hydroxyethyl)-l,2,4-oxadiazol-5-yl]pyrrolo[3,2-b]carbazole 2-(3-Ethyl- 1,2,4 -oxadiazol-5-yl)4— methyl- 1H-[1 ]benzofuro[2,3-/]indole 3,4-Dimethyl-2- 4,4-dimethyl-2-oxazolin-2-yl)pyrrolo[3,2-b]carbazole 3,4-Dimethyl-2- 3-(3-hydroxyphenyl)-l,2,4-oxadiazol-5-yl]pyrrolo[3,2- b]carbazole 3,4-Dimethyl-2- 3 -(N,N-dimethylaminomethy 1)- 1 ,2 ,4-oxadiazol-5 -yl]pyrrolo [3,2- b]carbazole 3,4-Dimethyl-2- tetrazol-5-yl)pyrrolo[3 ,2-b]carbazole) 3,4-Dimethyl-2- 3-(4-morpholinomethyl)-l,2,4-oxadiazol-5-yl)pyrrolo[3,2- b]carbazole 3,4-Dimethyl-2 3-methoxyethyl- 1 ,2,4-oxadiazol-5-yl)pyrrolo[3,2-b]carbazole 3,4-Dimethyl-2 l,2,4-oxadiazol-5-yl)pyrrolo[3,2-b]carbazole

and salts and physiologically functional derivatives thereof.

Compounds in this fourth series may be prepared according to the reaction schemes and procedures described in published International Patent Application No. WO96/01827, incorporated herein by reference.

According to a further aspect of the present invention there is provided use of a compound of formula (1) as hereinbefore defined in the manufacture of a medicament for use as a DHODH inhibitor.

According to a further aspect of the present invention there is provided a method of treating a patient requiring immunomodulation comprising administering to said patient an effective dose of a compound of formula (1) as defined above.

As used herein immunomodulation may comprise immunopotentiation or immunosuppression, preferably immunosuppression

According to a further aspect of the present invention there is provided a method of treating a patient requiring anti-inflammatory treatment comprising administering to said patient an effective dose of a compound of formula (1) as defined above.

According to a further aspect of the present invention there is provided a method of treating a patient having a condition in which inhibition of DHODH would be beneficial comprising administering to said patient an effective dose of a compound of formula (1) as defined above.

Conditions in which inhibition of DHODH is beneficial include: allergy, atopic dermatitis, urticaria, asthma, psoriasis, fibrosis, uveitis, rhinitis, colitis, SLE, autoimmune disease, cystic fibrosis, transplant rejection, graft-v-host disease, non- insulin dependent diabetes, multiple sclerosis, rheumatoid arthritis, sepsis, and parasitic infections such as protozoal infections including malaria, leishmaniasis and trypanosomiasis.

In the treatment of malaria, the compounds of the present invention may be combined with other anti-malarial agents such as proguanil.

According to a further aspect of the present invention there is provided a method of inhibiting DHODH in a patient comprising administering to said patient an effective dose of a compound of formula (1) as defined above.

Preferably the patient is a mammal; more preferably a human.

The amount of a compound of formula (1) required to be effective will, of course, vary and is ultimately at the discretion of the medical or veterinary practitioner. The factors to be considered include the condition being treated, the route of administration, the nature of the formulation, the mammal's body weight, surface area, age and general condition, and the particular compound to be administered. A suitable effective dose is in the range of about 0.01 to about 100 mg/kg body weight, eg 0.1 to about 100 mg/kg body weight, preferably 1-30 mg/kg body weight. The total daily dose may be given as a single dose, multiple doses, e.g., two to six times per day or by intravenous infusion for selected duration. For example, for a 75 kg mammal, the dose range would be about

8 to 900 mg per day, and a typical dose could be about 50 mg per day. If discrete multiple doses are indicated treatment might typically be 15 mg of a compound of formula (I) given up to 4 times per day.

Whilst it is possible for the active compound to be administered alone, it is preferable to present the active compound in a pharmaceutical formulation. Formulations of the present invention, for medical use, comprise a compound of formula (1) or a salt or physiologically functional derivative thereof together with one or more pharmaceutically acceptable carriers and optionally other therapeutic ingredients. The carrier(s) should be pharmaceutically acceptable in the sense of being compatible with the other ingredients of the formulation and not deleterious to the recipient thereof.

Formulations according to the present invention include those suitable for oral, topical, rectal or parenteral (including subcutaneous, intramuscular and intravenous) administration. Preferred formulations are those suitable for oral or parenteral administration.

The formulations may conveniently be presented in unit dosage form and may be prepared by any of the methods well known in the art of pharmacy. All methods include the step of bringing the active compound into association with a carrier which constitutes one or more accessory ingredients. In general, the formulations are prepared by uniformly and intimately bringing the active compound into association with a liquid carrier or a finely divided solid carrier or both and then, if necessary, shaping the product into desired formulations.

Formulations of the present invention suitable for oral administration may be presented as discrete units such as capsules, cachets, tablets or lozenges, each containing a predetermined amount of the active compound; as a powder or granules; or a solution or suspension in an aqueous or non-aqueous liquid such as a syrup, an elixir, an emulsion or a draught.

A tablet may be made by compression or moulding, optionally with one or more accessory ingredients. Compressed tablets may be prepared by compressing in a suitable machine the active compound in a free-flowing form such as a powder or granules, optionally mixed with a binder, lubricant, inert diluent, surface active or dispersing agent. Moulded tablets may be made by moulding in a suitable machine a mixture of the powdered active compound with any suitable carrier.

A syrup may be made by adding the active compound to a concentrated, aqueous solution of a sugar, for example sucrose, to which may also be added any accessory ingredients. Such accessory ingredients(s) may include flavourings, an agent to retard crystallisation of the sugar or an agent to increase the solubility of any other ingredients, such as a polyhydric alcohol for example glycerol or sorbitol. Formulations for rectal administration may be presented as a suppository with a conventional carrier such as cocoa butter.

Formulations suitable for parenteral administration conveniently comprise a sterile aqueous preparation of the active compound which is preferably isotonic with the blood of the recipient. Such formulations suitably comprise a solution of a pharmaceutically and pharmacologically acceptable acid addition salt of a compound of the formula (1) that is isotonic with the blood of the recipient.

Useful formulations also comprise concentrated solutions or solids containing the compound of formula (1) which upon dilution with an appropriate solvent give a solution for parenteral administration as above.

In addition to the aforementioned ingredients, the formulations of this invention may further include one or more accessory ingredient(s) selected from diluents, buffers, flavouring agents, binders, surface active agents, thickeners, lubricants, preservatives (including antioxidants) and the like.

The invention will now be described with reference to the following non-limitative Examples and Figures in which:-

Figure 1 illustrates inhibition of DHODH activity by ethyl 3, 4-dimethylpyrrolo [3,2,- b]carbazole-2-carboxylate (Example 1) in crude lysate mitochondrial preparations using a dichlorophenol-indophenol coupled assay.

Figure 2 illustrates the effect of the compound of Example 1 on anti-CD3 induced proliferation of PBMC at cell concentrations a) 1.5 x 10⁵ cells/well; b) 7.5 x 10⁴ cells/well; c) 3.75 x 10⁴ cells/well; and d) 1.87 x 10⁴ cells/well. Open circles represent day one; solid circles represent day 2; open squares represent day 3; and solid squares represent day 4.

Figure 3 illustrates the effect on cell cycle distribution (10⁶ cells) over seven days of a) no stimulation; b) anti-CD3 stimulation; c) compound stimulation; and d) anti-CD3 and compound stimulation. Open squares represent Gl phase; open circles represent S phase; solid circles represent G2.M phase. Figure 4 illustrates the effect of the compound of Example 1 on PPd induced proliferation of PBMC. Open circles relate to no PPd; solid circles relate to PPd at 10 μg/ml;

Figure 5a illustrates the effect of the compound of Example 1 on anti-CD3 induced proliferation of PBMC. Solid squares represent medium control experiment; solid circles represent anti-CD3 at 100 ng/ml; triangles represent anti-CD3 at 100 ng/ml along with the compound at lμM; and inverted triangles represent anti-CD3 at 100 ng/ml along with the compound at lOμM.

Figure 5b illustrates the effect of the compound of Example 1 on PPd induced proliferation of PBMC. Solid square represent medium control experiment; solid circles represent PPd at lOμg/ml; triangles represent PPd at lOμg/ml along with the compound at lμM; and inverted triangles represent PPd at lOμg/ml along with the compound at lOμM.

The following examples are provided by way of example only. It will be appreciated that modification of detail may be made without departing from the scope of the invention.

Example 1 - Ethyl 3,4-Dimethyl[3,2,b]carbazole-2-carboxylate

Synthesis of this compound is described in published International Patent Application WO94/02483.

One-pot Synthesis of the Pyrrolocarbazoles - General procedure

A solution of indole (1.0 mmol) and the 5-acetoxymethyl-4-acetylpyrrole (1.0 mmol) in 1 ,2-dichloroethane (10 cm³) was heated under gentle reflux and stirred with Montomorillonite K10 clay (1 g) for 3-4 h. The colour of clay turned light brown and the reaction was followed to completion by TLC. After filtration from clay and washing well with 1 ,2-dichloroethane, evaporation of the combined filtrates gave the pyrrolo[3,2-b]carbazoles which were obtained as yellow crystals after crystallisation from dichloromethane or ethyl acetate.

Ethyl 3,4-dimethylpyrrolo[3,2,-b]carbazole-2-carboxylate

(0.199 g, 65%) was obtained from the reaction of indole and the 5-acetoxymethyl-4- acetylpyrrole.

Assay for DHODH Inhibitory Activity

DHODH catalyses the oxidation of dihydroorotate to orotic acid and is the fourth enzyme of the de novo pyrimidine biosynthesis pathway, residing on the outer surface of the inner mitochondrial membrane (Jones, M.E. Pyrimidine nucleotide biosynthesis in animals: genes, enzymes, and regulation of ump biosynthesis. Annual Review of Biochemistry, 49:253-79, 1980. ,35:253-279, 1980.)

DHODH activity was measured in mitochondrial preparations. Crude mitochondrial preparations were obtained as follows: subconfluent DLD-1 cells were harvested and snap-frozen at -70C in PBS. After thawing, cells were washed and resuspended in 0.25M sucrose, lmM EGTA and lOmM Hepes/NaOH pH7.0. Cells were then homogenised using 25 strokes in a Dounce homogeniser with a tight pestle. Nuclei were pelleted at 1500 x g and a crude mitochondrial pellet was obtained by subsequent centrifugation of the supernatant at 10000 x g. Dihydroorotate dehydrogenase activity was measured in the crude lysates using a dichlorophenol-indophenol coupled assay (Lakaschus, G. and Loffler M. Differential susceptibility of dihydroorotate dehydrogenase/oxidase to brequinar sodium (nsc 368 390) in vitro. Biochemical Pharmacology, 43: 1025-1030, 1992.)

Results

It was found that the compound of Example 1 inhibited DHODH (see Figure 1) with an IC₅₀ of0.4μM. Immune Cell Function

The effect of the compound of Example 1 on polyclonal (anti-CD3) stimulation of PBMC has been studied. The immunopotentiation effect of 1 μM of the compound is greatest at low cell concentrations (1.8 x 10⁴ cells/well) and results in a 5-fold increase in proliferation above background levels (Figure 2).

The effect of the compound on anti-CD3 activation of high and low cell concentrations of PBMC was studied. The effect on the cell cycle distribution of cells for up to 7 days following activation was examined and the expression of CD25 on CD4 and CD8 +ve T-cells over the same time period monitored (Figure 3). At high and low density <15% cells accumulated in S phase by day 3-4 in the absence of stimulation, this was elevated to approx 20% in the presence of the compound or anti-CD3 alone. At high cell densities in the presence of anti-CD3 and the compound, approximately 50% of the cells were in S phase by day 4. At low cell densities this figure was reduced to approx 35%. There appeared to be a greater number of blast cells at the lower cell densities. The compound had little effect on the expression of CD25 on CD4 and CD8 +ve T-cells.

The effect of the compound of Example 1 on antigen (PPd) specific stimulation of PBMC has been studied (Figure 4). The compound was found to exhibit a dose dependent effect on PPd induced proliferation. At lOμM the compound of Example 1 was strongly inhibitory of PPd induced proliferation.

To establish whether a dose of lOμM was toxic to PBMC, the kinetics of the response to PPd and anti-CD3 in the presence of the compound at 10 and lμM was studied. lOμM was not toxic to PBMC in that they were able to mount an effective response to anti-CD3 although that response was less than that seen in the presence of lμM or its absence. The presence of the compound may prolong the proliferation of anti-CD3 induced cells possibly by preventing the 'burn-out' of proliferating cells. lμM enhanced proliferation to anti-CD3. In contrast, lOμM almost completely inhibited the proliferation of PBMC to PPd whereas lμM had little effect. Therefore lOμM of the compound of Example 1 is not toxic to immune cells but this dose has a profound inhibitory effect on the immunocompetence of cells in response to an antigen specific stimulation.

Claims

CLAIMS:

1. Use of a compound of formula (1) in the manufacture of a medicament for use as an immunomodulatory or anti-inflammatory drug or for use in the treatment of a therapeutic indication in which inhibition of dehydro-orate dehydrogenase (DHODH) is beneficial:-

and salts and physiologically functional salts thereof,

R⁵ and R⁶ are independently selected from H, hydroxy, nitro, amino, halo, cyano, CHO, COR⁸, CO₂R⁸ and the following groups which may be optionally substituted: alkyl, aryl, aryloxy, aralkyloxy, alkoxy, aralkyl, wherein R is optionally substituted alkyl and aryl;

A is:

wherein R¹ is COR⁸, CHO, CH₂OH, CH₂OR⁸, CONH₂, COOR⁸, CONHR⁸, CONR⁸R⁸, 8 , 8╧ä> 8

CONHNR > 8⁶╧äR> ^╬┤, COO(CH₂)_nNR^╬┤R^╬┤, CSOR⁸, CSSR⁶, COSR⁸, CSNHR⁸, CSNR 8^╬┤ ╧ä R8⁸, CNHOR⁸ or an optionally substituted 5 or 6 membered aromatic or nonaromatic heterocyclic ring containing 1 to 4 heteroatoms,

wherein the groups R⁸ are independently selected from hydrogen, optionally substituted alkyl, aryl, aralkyl, acyl, alkoxyalkyl, heterocycloalkyl and heteroaralkyl groups, and Ci-io optionally substituted hydrocarbyl groups which may contain one or two oxygen atoms in the chain and wherein n is 1 to 4;

or R may independently be sugar groups;

R² is H, hydroxy, haloalkyl, halo, cyano, COOR⁸, alkyl, aryl, alkenyl, alkynyl, alkoxy, (wherein alkyl, aryl, alkenyl, alkynyl and alkoxy can be substituted) CH₂CH₂CO₂R⁹ (wherein R⁹ is alkyl or aryl), CHO, COR⁸, COOR⁸ or a C_MO optionally substituted hydrocarbyl group which may contain one or two oxygen atoms in the chain, wherein R⁸ is independently selected from the groups defined for R⁸ above;

Y is O, S, SO, SO₂, CH₂, CO or NR⁷ wherein R⁷ is independently selected from groups hereinbefore defined for R⁷;

Z and Z are independently selected from H, halogen, cyano, amino, alkyl, COOR , CONHR⁸, COR⁸, CH₂OH, CH₂OR⁸, CONH₂, CON R⁸R⁸, CSOR⁸, CSSR⁸, COSR⁸, CSNHR⁸, CSNR⁸R⁸ and CNHOR⁸ wherein R⁸ is independently selected from the groups defined for R⁸ above; or Z¹ and Z² together form the group:

wherein R³ and R⁴ are independently selected from H, hydroxy, alkyl, haloalkyl, alkoxy, halo, cyano, azido, nitro, amino, alkyl amino, dialkyl amino, CHO, COR⁸, CONHR⁸, CON R⁸R⁸ (wherein R⁸ is independently selected from the groups defined for R⁸ above ),carboxyl or CO₂R¹⁰, wherein R¹⁰ is independently selected from alkyl, aralkyl and aryl.

2. Use of a compound according to claim 1, wherein Z¹ and Z² together form the group:

wherein R³ and R⁴ are independently selected from H, hydroxy, alkyl, haloalkyl, alkoxy, halo, cyano, azido, nitro, ammo, alkyl ammo, dialkyl amino, CHO, COR , CONHR⁸, CON R⁸R⁸ (wherein R⁸ is independently selected from the groups defined for R⁸ above ),carboxyl or CO₂R¹⁰, wherein R¹⁰ is independently selected from alkyl, aralkyl and aryl.

3. Use of a compound according to claim 2, wherein

X is O, S, SO, SO₂, CH₂, CO or NR⁷, wherein R⁷ is H, alkyl, aralkyl, aryl, alkenyl, acyl, alkynyl, sulphonyl or substituted sulphonyl; and

Y is O, S, SO, SO₂, CH₂, CO or NR⁷.

4. Use of a compound according to claim 3 wherein the compound is of formula (II)

Y is O, S, SO, SO₂, CH₂, CO or NR⁷;

R³ and R⁴ are independently H,hydroxy, alkyl, haloalkyl, alkoxy, halo, cyano, nitro, amino, alkyl amino, dialkyl amino, substituted alkyl, carboxyl or CO₂R⁹;

Use of a compound according to claim 4 wherein the compound is selected from

3-Pyridyl 3, 4-dimethylpyrrolo( 3, 2-b] carbazole-2-carboxylate [(3-Dimethylamino)phenyl]3, 4-dimethylpyrrolo[3,2-b] carbazole-2-carboxylate Benzyl 1, 3, 4- trimethylpyrrolo( 3, 2-b] carbazole-2-carboxylate Phenyl 3, 4-dimethylpyrrolo [ 3, 2-b] carbazole-2-carboxylate 3,4-Dimethyl-2-( 1-imidazolylcarbonyl ) pyrrolo [3, 2-b] carbazole Ethyl 3, 4-dimethylpyrrolo [3,2,-b]carbazole-2-carboxylate; Ethyl 3, 4-dimethylbenzothieno [4,

5-f] indole-2-carboxylate; Benzyl 3, 4-dimethylpyrrolo [3, 24)] carbazole-2-carboxylate; Benzyl 8-f luoro-3, 4-dimethylpyrrolo [3,2-b] carbazole-2-carboxylate; Ethyl 8-f luoro-3, 4-dimethylpyrrolo [3,2-b] carbazole-2-carboxylate Benzyl 3,4, 6-trimethylpyrrolo (3, 2-b] carbazole-2-carboxylate;

Ethyl 3,4, 6-trimethylpyrrolo [3, 2-b] carbazole-2-carboxylate;

8-Fluoro-3, 4-dimethylpyrrolo[ 3, 2-b] carbazole-2-carboxylic acid

3,4-Dimethylpyrrolo[ 3, 2-Jb] carbazole-2-carboxylic acid;

Ethyl 8-methoxy-3, 4-dimethylpyrrolo [3, 2-b] carbazole-2-carboxylate;

3,4,6-Trimethylpyrrolo [3, 2-b] carbazole-2-carboxylic acid and

Benzyl 8-methoxy-3, 4-dimethylpyrrolo [3, 2-_b] carbazole-2-carboxylate;

and physiologically functional derivatives thereof.

6. Use of a compound according to claim 1 wherein the compound is ethyl 3,4- dimethylpyrrolo [3,2-b]carbazole-2-carboxylate or a physiologically functional derivative thereof.

7. Use of a compound according to claim 1 wherein the compound is of the formula (III)

X is O, S, SO, SO₂, CH₂, CO or NR⁷, wherein R⁷ is H, alkyl, aralkyl, aryl, alkenyl, acyl, alkynyl, sulphonyl or substituted sulphonyl; Y is O, S, SO, SO₂, CH₂, CO or NR⁷;

R¹ is COOR¹⁶, CONHR¹⁶, CONR¹⁶R¹⁷, CSOR¹⁶, CSSR¹⁶, COSR¹⁶, CSNHR¹⁶, CSNR¹⁶R¹⁷, CNHOR¹⁶ wherein R¹⁶ and R¹⁷ are independently C_1-10 optionally substituted hydrocarbyl groups which may optionally contain one or two oxygen atoms in the chain; or R¹⁶ and R¹⁷ are independently alkoxyalkyl, heterocycloalkyl, heteroaralkyl,

or R¹⁶ and R¹⁷ may independently be sugar groups;

R² is H, halo, cyano, COOR¹⁶, alkyl, aryl, alkenyl, alkynyl, alkoxy, (wherein alkyl, aryl, alkenyl, alkynyl and alkoxy can be substituted) or CH₂CH₂CO₂R⁹ wherein R⁹ is alkyl or aryl;

R³ and R⁴ are independently H,hydroxy, alkyl, haloalkyl, alkoxy, halo, cyano, nitro, amino, alkyl amino, dialkyl amino, substituted alkyl, carboxyl or CO R⁹;

8. Use of a compound according to claim 1 wherein the compound is selected

[(2-Dimethylamino)ethyl] 3,4-dimethylpyrrolo[3,2-b]carbazole-2-carboxylate (2-Methylsulphonylethyl) 3,4-dimethylpyrrolo[3,2-b]carbazole-2-carboxylate (2-Methylsulphinylethyl) 3,4-dimethylpyrrolo[3,2-b]carbazole-2-carboxylate (l,3-Dibenzyloxypropyl-2) 3,4-dimethylpyrrolo[3,2-b]carbazole-2-carboxylate (l-Benzyloxy-3-hydroxypropyl-2) 3,4-dimethylpyrrolo[3,2-b]carbazole-2-carboxylate ( 1 ,3-Dihydroxypropyl-2) 3 ,4-dimethyl-pyrrolo [3 ,2-b] carbazole-2-carboxylate (2- Amino-2-methylpropyl- 1) 3 ,4-dimethylpyrrolo[3 ,2-b] carbazole-2-carboxylate (4-Nitrophenylmethyl) 2-(3 ,4-dimethylpyrrolo [3 ,2-b]carbazole-2-carboxy)acetate 2-(3,4-Dimethylpyrrolo[3,2-b]carbazole-2-carboxy)acetic acid Cyclohexyl 3,4-dimethylpyrrolo[3,2-b]carbazole-2-carboxylate Cyclohexylmethyl 3,4-dimethylpyrrolo[3,2-b]carbazole-2-carboxylate Cyclopentyl 3,4-dimethylpyrrolo[3,2-b]carbazole-2-carboxylate Cyclooctyl 3,4-dimethylpyrrolo[3,2-b]carbazole-2-carboxylate

3,5-Dihydroxycyclohexyl 3,4-dimethylpyrrolo[3,2-b]carbazole-2-carboxylate cis-4-tert-Butyldiphenylsilyloxycyclohexyl 3,4-dimethylpyrrolo[3,2-b]carbazole-2- carboxylate cis-4-Hydroxycyclohexyl 3,4-dimethylpyrrolo[3,2-b]carbazole-2-carboxylate trans-4-tert-Butyldiphenylsilyloxycyclohexyl 3,4-dimethylpyrrolo[3,2-b]carbazole-2- carboxylate trans4-Hydroxycyclohexyl 3,4-dimethylpyrrolo[3,2-b]carbazole-2-carboxylate

Tetrahydro-2H-pyran4-yl 3,4-dimethylpyrrolo[3,2-b]carbazole-2-carboxylate l-Benzylpiperidin-4-yl 3,4-dimethylpyrrolo[3,2-b]carbazole-2-carboxylate Piperidin-4-yl 3 ,4-dimethylpyrrolo [3,2-b] carbazole-2-carboxylate l-Methylpiperidin-4-yl 3,4-dimethylpyrrolo[3,2-b]carbazole-2-carboxylate (3,4-Dimethylpyrrolo[3,2-b]2-carbazolyl)3-O-( l,2:5,6-di-O-isopropylidene glucofuranoside) (3,4-Dimethylpyrrolo[3,2-b]2-carbazolyl) 3-O-(l ,2-O-isopropyl-ideneglucofuranoside) [3-(4-Nitrophenylmethoxy)phenyl] 3,4-dimethylpyrrolo[3,2-b]carbazole-2-carboxylate (3-Hydroxyphenyl) 3-4,-dimethylpyrrolo[3,2-b]carbazole-2-carboxylate (4-Phthalamidophenyl) 3,4-dimethylpyrrolo[3,2-b]carbazole-2-carboxylate 4-(Aminophenyl) 3, 4-dimethylpyrrolo [3 ,2-b]carbazole-2-carboxylate (4-Nitrophenylmethyl) 3-(3,4-dimethylpyrrolo[3,2-b]carbazole-2-carboxy)benzoate

(3-Hydroxymethyl)phenyl 3 ,4-dimethylpyrrolo[3,2-b]carbazole-2-carboxylate [3-(4-Nitrophenylmethoxy)phenyl] 4-methyl-lH-[l]benzothieno[2,3 -f]indole-2- carboxylate

(3-Hydroxyphenyl)4-methyl lH-[ 1 ]benzothieno[2,3-f]indole-2-carboxylate 3-(4-Nitrophenylmethoxy)phenylmethyl 3,4-dimethylpyrrolo[3,2-b]carbazole-2- carboxylate 3-(tert-Butyldiphenylsilyloxyphenyl)methyl 3,4-dimethylpyrrolo[3,2-b]carbazole-2- carboxylate

(3-Hydroxyphenyl)methyl 3 ,4-dimethylpyrrolo[3,2-b]carbazole-2-carboxylate ( 1 -Hydroxy-3 -methylpropyl-2) 3 ,4-dimethylpyrrolo[3 ,2-b] carbazole-2-carboxamide 2-Hydroxyethyl 3,4-dimethylpyrrolo[3,2-b]carbazole-2-carboxamide 2-Chloroethyl 3,4-dimethylpyrrolo[3,2-b]carbazole-2-carboxamide N-(2-Aminoethyl) 3, 4-dimethylpyrrolo [3, 2-b] carbazole-2-carboxamide N-(2-Acetamidoethyl) 3,4-dimethylpy╧ÇOlo[3,2-b]carbazole-2-carboxamide (3-Aminopropyl- 1) 3,4-dimethylpyrrolo[3,2-b]carbazole-2-carboxamide 2-Hydroxyethyl 4-methyl- 1H-[1 ]benzothieno[2,3-f]indole-2-carboxamide 2-Chloroethyl 4-methyl-lH-[l]benzothieno[2,3-f]indole-2-carboxamide and physiologially functional derivatives thereof.

9. Use of a compound according to claim 1 wherein the compound is of the formula (V)

Y is O, S, SO, SO₂, CH₂, CO or NR⁷;

R¹ is COR¹⁸, CHO, CH₂OH, CH₂OR¹⁸, CONH₂, COOR¹⁸, CONHR¹⁸, CONR¹⁸R¹⁹, CSOR¹⁸, CSSR¹⁸, COSR¹⁸, CSNHR¹⁸, CSNR¹⁸R¹⁹, CNHOR¹⁸ wherein R¹⁸ and R¹⁹ are independently hydrogen, alkoxyalkyl, heterocycloalkyl, heteroaralkyl, or C_MO optionally substituted hydrocarbyl group which may optionally contain one or two oxygen atoms in the chain;

or R¹⁸ and R¹⁹ may independently be sugar groups;

Z¹ is H, alkyl, halogen, cyano, amino, COOR¹⁸, CONHR¹⁸, COR¹⁸, CH₂OH, CH2OR¹⁸, CONH₂, CONR¹⁸R¹⁹, CSOR¹⁸, CSSR¹⁸, COSR¹⁸, CSNHR¹⁸, CSNR¹⁸R¹⁹ or CNHOR¹⁸; Z² is H, halogen, cyano, amino, alkyl, COOR¹⁸, CONHR¹⁸, COR¹⁸, CH₂OH, CH₂OR¹⁸, CONH₂, CONR¹⁸R¹⁹, CSOR¹⁸, CSSR¹⁸, COSR¹⁸, CSNHR¹⁸, CSNR¹⁸R¹⁹ or CNHOR¹⁸;

10. Use of a compound according to claim 9 wherein the compound is selected from:-

Ethyl l,7-dihydro-3,4,6-trimethylpyrrolo[3,2-f]indole-2-carboxylate; Diethyl l,7-dihydro-3,4,6-trimethylpyrrolo[3,2-f] indole-2,5-dicarboxylate; and Ethyl 6-methoxycarbonyl-3 , 4-dimethylpyrrolo [3 ,2-f]indole-2-carboxylate

and physiologically functional derivatives thereof; and

Ethyl 6-Benzyloxycarbonyl-3,4-dimethylpyrrolo[3,2-f ]indole-2-carboxylate; Dibenzyl 3,4-dimethylpyrrolo[3,2-f]indole-2,6-dicarboxylate; Ethyl 7-methoxycarbonyl-3,4-dimethylpyrrolo [3,2-f]indole-2-carboxylate; and Ethyl 3,4-dimethylpyrrolo[3,2-f]indole-2-carboxylate and physiologically functional derivatives thereof.

11. Use of a compound according to claim 1 wherein the compound is of the formula (VI)

and salts and physiologically functional derivative thereof, wherein B is

Y is O, S, SO, SO₂, CH₂, CO or NR⁷;

R¹ is an optionally substituted 5- or 6-membered heterocyclic ring containing 1 to 4 heteroatoms wherein the 5- or 6- membered ring may be aromatic or non-aromatic;

R² is H, hydroxy, halo, haloalkyl, cyano, alkyl, aryl, alkenyl, alkynyl, alkoxy, (wherein alkyl, aryl, alkenyl, alkynyl, and alkoxy can be substituted), CHO, COR²³, COOR²³ wherein R²³ is hydrogen or is a C _O optionally substituted hydrocarbyl group which may contain one or two oxygen atoms;

R ,21 i ΓÇós H, hydroxy, nitro, amino, halo, cyano, CHO, COR 23 , or the following groups which may be optionally substituted: alkyl, aryl, aryloxy, aralkyloxy, alkoxy, aralkyl;

12. Use of a compound according to claim 11 wherein the compound is selected from:- 3,4-Dimethyl-2-(3-ethyl-l,2,4-oxadiazol-5-yl) pyrrolo[3,2-b]carbazole;

2-(3-Benzyl- l,2,4-oxadiazol-5-yl)-3,4-dimethylpyrrolo[3,2-b]carbazole;

3,4-Dimethyl-2-(3-ethyl-l,2,4-oxadiazol-5-yl) pyrrolo[2,3-b]carbazole;

2-(3-ethyl-l,2,4-oxadiazol-5-yl)-4-methyl-lH-[l]benzothieno[2,3-/]indole; 3,4-Dimethyl-2-(2-methyl-l,3,4-oxadiazol-5-yl)-pyrrolo[3,2-/3]carbazole;

3,4-Dimethyl-2-(2-ethyl-l,3,4-oxadiazol-5-yl)-pyrrolo[3,2-/3]carbazole;

3,4-Dimethyl-2-(2-phenyl-l,3,4-oxadiazol-5-yl)-pyrrolo[3,2-b]carbazole;

2-(2-Ethyl-l,3,4-oxadiazol-5-yl)4-methyl-lH-[l]benzothieno[2,3-/]indole;

3,4-Dimethyl-2-[3-(3,4-methylenedioxyphenyl)-l,2,4-oxadiazol-5-yl]pyrrolo[3,2- /yjcarbazole;

4-Methyl-2-(2-oxazolin-2-yl)- 1 H-[ 1 ]benzothieno[2,3 -/Jindole

3,4-Dimethyl-2-(3-methyl-l,2,4-oxadiazol-5-yl)pyrrolo[3,2-/ ]carbazole

3 ,4-Dimethy l-2-(3 -methyl- 1 ,2 ,4-oxadiazol-5 -yl)pyrrolo [2,3 -b] carbazole

3,4-Dimethyl-2-[(3-phenyl)-l,2,4-oxadiazol-5-yl]pyrrolo[3,2-/3]carbazole 3,4-Dimethyl-2-(2-oxazolin-2-yl)pyrrolo[3,2-b]carbazole

3,4-Dimethyl-2-[3-(l-piperidinylmethyl)-l,2,4-oxadiazol-5-yl)pyrrolo[3,2- b]carbazole

3,4-Dimethyl-2-[3-(4-pyridyl)-l,2,4-oxadiazol-5-yl]pyrrolo[3,2-b]carbazole

3,4-Dimethyl-2-(3-methoxymethyl-l,2,4-oxadiazol-5-yl)pyrrolo[3,2-b]carbazole 3,4-Dimethyl-2-[3-(2-hydroxyethyl)-l,2,4-oxadiazol-5-yl]╧üyrrolo[3,2-b]carbazole

2-(3-Ethyl- l,2,4-oxadiazol-5-yl)4~methyl- lH-[l]benzofuro[2,3-/]indole

3,4-Dimethyl-2-(4,4-dimethyl-2-oxazolin-2-yl)pyrrolo[3,2-/3]carbazole

3,4-Dimethyl-2-[3-(3-hydroxyphenyl)-l,2,4-oxadiazol-5-yl]pyrrolo[3,2- b] carbazole 3,4-Dimethyl-2-[3-(N,N-dimethylaminomethyl)-l,2,4-oxadiazol-5-yl]pyrrolo[3,2- b]carbazole

3,4-Dimethyl-2-(tetrazol-5-yl)pyrrolo[3 ,2-b]carbazole)

3 ,4-Dimethy 1-2- [3 -(4-morpholinomethyl)- 1 ,2 ,4-oxadiazol-5 -yl)pyrrolo [3 ,2- b]carbazole 3,4-Dimethyl-2-(3-methoxyethyl- l,2,4-oxadiazol-5-yl)pyrrolo[3,2-b]carbazole

3,4-Dimethyl-2-(l,2,4-oxadiazol-5-yl)pyrrolo[3,2-b]carbazole

and salts and physiologically functional derivatives thereof.

13. Use of a compound as defined in any one of claims 1 to 12 in the manufacture of a medicament for use as an immunomodulating drug.

14. Use of a compound as defined in any one of claims 1 to 12 in the manufacture of a medicament for use as an immunosuppressive drug.

15. Use of a compound as defined in any one of claims 1 to 12 in the manufacture of a medicament for use as an anti-inflammatory drug

16. Use of a compound as defined in any one of claims 1 to 12 in the manufacture of a medicament for use in the treatment of a therapeutic indication in which inhibition of DHODH is beneficial.

17. Use of a compound as defined in any one of claims 1 to 12 in the manufacture of a medicament for use as a DHODH inhibitor.

18. A method of treating a patient requiring immunomodulation comprising administering to said patent an effective dose of a compound as defined in any one of claims 1 to 12.

19. A method of treating a patient requiring immunosuppression comprising administering to said patent an effective dose of a compound as defined in any one of claims 1 to 12.

20. A method of treating a patient requiring anti-inflammatory treatment comprising administering to said patent an effective dose of a compound as defined in any one of claims 1 to 12.

21. A method of treating a patient having a condition in which inhibition of DHODH would be beneficial comprising administering to said patent an effective dose of a compound as defined in any one of claims 1 to 12.

22. A method of inhibiting DHODH in a patient comprising administering comprising administering to said patent an effective dose of a compound as defined in any one of claims 1 to 12.