Derivatives of indole and their use as antagonists of mcp-1

 

(57) Abstract:

The invention relates to indole derivative of General formula I:

where R1represents hydrogen, halogen, methoxy; R2represents hydrogen, halogen, methyl, ethyl, methoxy; R3represents carboxy, tetrazolyl or CONHSO2R4in which R4represents methyl, ethyl, phenyl, 2,5-dimethylisoxazole, trifluoromethyl; T represents-CH2- or-SO2-; and ring a is 3-chlorophenyl, 4-chlorophenyl, 3-triptoreline, 3,4-dichlorophenyl, 3,4-differenl, 3-fluoro-4-chlorophenyl, 3-chloro-4-forfinal, 2,3-dichloride-5-yl; or their pharmaceutically acceptable salts or esters, as well as pharmaceutical compositions containing them. These compounds and compositions can be used in the treatment of diseases mediated macrophage chemoattractant protein-1 or RANTES (regulated upon activation, expressed and secretory normal T-cells), such as an inflammatory disease. 3 N. and 7 C.p. f-crystals, 1 table.

The present invention relates to anti-inflammatory compounds that act via antagonism of the receptor CCR2 (also known as the receptor for MCP-1), leading, narrat indole fragment. The invention also relates to the containing pharmaceutical compositions, methods for their preparation, intermediate compounds useful for their production and to their use as therapeutic agents.

MCP-1 is a member of the family of chemokines proinflammatory proteins, which mediate the chemotaxis and activation of leukocytes. MCP-1 is a C-C chemokine, which is one of the most potent and selective of the famous chemoattractant and activating means of the T-cells and monocytes. MCP-1 is involved in the pathophysiology of a huge number of inflammatory diseases, including rheumatoid arthritis, glomerulonephritis, pulmonary fibrosis, restenosis (international patent application WO 94/09128), alveolitis (Jones et al., 1992, J. Immunol., 149, 2147) and asthma. Other pathological processes in which, say, MCP-1 plays a role in their pathology is atherosclerosis (e.g., Koch et al., 1992, J. Clin. Invest., 90, 772-779), psoriasis (Deleuran et al., 1996, J. Dermatological Science, 13, 228-236), skin reactions of delayed-type hypersensitivity, inflammatory bowel disease (Grimm et al., 1996, J. Leukocyte Biol., 59, 804-812), multiple sclerosis and traumatic brain injury (Berman et al., 1996, J. Immunol., 156, 3017-3023). Inhibitor of MCP-1 may be useful for the treatment and acts through the receptor CCR2. MCP-2 and MCP-3 can also, at least in part, act through this receptor. Therefore, when this description refers to “the inhibition or antagonism of MCP-1 or the effects mediated MCP-1”, this includes the inhibition or antagonism in relation to effects mediated MCP-2 and/or MCP-3, when MCP-2 and/or MCP-3 function via receptor CCR2.

Applicants have discovered a class of compounds containing the indole fragment, which have a useful inhibitory activity against MCP-1. Are simultaneously pending application UK 9716657.3 discloses a class of indoles with inhibitory activity against MCP-1. This application is based on the surprising discovery that specific substituted 5-hydroxyindole are inhibitors MPC-1, which have unexpected and favorable properties in terms of power and/or blood levels and/or bioavailability and/or solubility.

Accordingly, the present invention provides compounds of formula (I):

in which

R1represents hydrogen, halogen or methoxy;

R2represents hydrogen, halogen, methyl, ethyl or methoxy;

R3present oxazolyl or trifluoromethyl;

T represents-CH2- or-SO2; and

ring a is 3-chlorophenyl, 4-chlorophenyl, 3-triptoreline, 3,4-dichlorophenyl, 3,4-differenl, 3-fluoro-4-chlorophenyl, 3-chloro-4-forfinal or 2,3-dichloride-5-yl; or their pharmaceutically acceptable salts or prodrugs.

In this description, the term “alkyl” includes alkyl groups with both straight and branched chain, but references to individual alkyl groups such as “propyl” are specific only for remotemachine options. The term “halogen” refers to fluorine, chlorine, bromine and iodine.

The particular new compounds of the invention include, for example, compounds of formula (I) or their pharmaceutically acceptable salts or prodrugs, in which in the absence of other indications:

a) R1has any of the values defined here below in paragraphs (i)-(iii), or a combination of two of these values;

b) R2has any of the values defined here below in paragraphs (iv)-(viii), or a combination of two of these values;

c) R3has any of the values defined here below in paragraphs (ix)-(xi), or a combination of two of these values;

e) T has any of the values defined here below in paragraphs xii)-his these values;

i) R1represents hydrogen;

ii) R1represents halogen;

iii) R1represents methoxy;

iv) R2represents hydrogen;

v) R2represents halogen;

vi) R2is methyl;

vii) R2represents ethyl;

viii) R2represents methoxy;

ix) R3represents carboxy;

x) R3is tetrazolyl;

xi) R3is-CONHSO2R4where R4represents methyl, ethyl, phenyl, 2,5-dimethylisoxazole or trifluoromethyl;

xii) T represents-CH2-;

xiii) T is-SO2-;

xiv) the ring a is 3-chlorophenyl;

xv) the ring a represents 4-chlorophenyl;

xvi) the ring a represents a 3-triptoreline;

xvii) the ring a is 3,4-dichlorophenyl;

xviii) the ring a is 3,4-differenl;

xix) the ring a is 3-fluoro-4-chlorophenyl;

XX) the ring a represents a 3-chloro-4-forfinal; and

xxi) the ring a represents a 2,3-dichloride-5-yl.

Preferably R1represents hydrogen.

Preferably R2is W is-CH2-.

Preferably ring a is 3-chlorophenyl, 4-chlorophenyl, 3-triptoreline, 3,4-dichlorophenyl, 3,4-differenl, 3-fluoro-4-chlorophenyl or 3-chloro-4-forfinal.

More preferably, ring a is 3,4-dichlorophenyl, 3-fluoro-4-chlorophenyl or 3-chloro-4-forfinal.

For example, ring a is 3,4-dichlorophenyl or 3-chloro-4-forfinal.

In another aspect of the invention preferably ring a is 3,4-dichlorophenyl, 2,3-dichloride-5-yl or 3-chloro-4-forfinal.

Therefore, in a preferred aspect of the invention provides compounds of formula (I) shown above in which:

R1represents hydrogen;

R2represents hydrogen;

R3represents carboxy;

T represents-CH2-; and

ring a is 3,4-dichlorophenyl, 3-fluoro-4-chlorophenyl or 3-chloro-4-forfinal; in particular, 3,4-dichlorophenyl or 3-chloro-4-forfinal; or its pharmaceutically acceptable salt or prodrug.

Preferred compounds of the invention include any compound examples. More preferred compounds of the invention are compounds of examples 1, 3 and 4, for example, example 1 and 3.

The compounds of formula (I) are inhibitors chemoattractant protein-1 monocytes. In addition, they apparently inhibit chemotaxis induced RANTES. RANTES (regulated upon activation, expressed and secretory normal T-cells) is another chemokine from the same family, and that MCP-1, with a similar biological profile, but acting through the receptor CCR-1. As a result, these compounds can be used for treating diseases mediated by these agents, in particular inflammatory diseases.

Suitable pharmaceutically acceptable salts of compounds of formula (I) include salts of bases, such as alkali metal salts, e.g. sodium, salts of alkaline earth metals, e.g. calcium or magnesium salt of an organic amine, such as triethylamine, research, N-methylpiperidine, N-ethylpiperidine, procaine, dibenzylamine, N,N-dibenzylamine or amino acids for example lysine. In another aspect, when coedine, fumarate, hydrochloride, hydrobromide, citrate, maleate and salts formed with phosphoric and sulphuric acid. There may be several cations or anions, depending on the number of charged features and valence of the cations or anions. Preferred pharmaceutically acceptable salt is a sodium salt.

In this area there are various forms of prodrugs. Examples of such derivatives, prodrugs can be found in the publications of:

a) Design of Prodrugs, edited by H. Bundgaard (Elsevier, 1985) and Methods in Enzymology, Vol.42, p.309-396, edited by K. Widder, et al. (Academic Press, 1985);

b) A Textbook of Drug Design and Development, edited by Krogsgaard-Larsen and H. Bundgaard, Chapter 5 "Design and Application of Prodrugs", by H. Bungaard, p.113-191 (1991);

c) H. Bungaard, Advanced Drug Delivery Reviews, 8, 1-38 (1992);

d) H. Bungaard et al., Journal of Pharmaceutical Sciences, 77, 285 (1988); and

e) N. Kakeya, et al., Chem Pharm Bull, 32, 692 (1984).

Examples of such prodrugs are cleaved in vivo esters of the compounds of the invention. Cleaved in vivo by a complex ester compounds of the invention containing carboxypropyl is, for example, pharmaceutically acceptable ester which is cleaved in the human or animal with the formation of the original acid. Suitable pharmaceutically acceptable esters for carboxypropyl involve complex is oximately; C1-6alkanoyloxy esters, for example, pivaloyloxymethyl; palidrome esters; C3-8cycloalkylcarbonyl-C1-6alkalemia esters, for example, 1-cyclohexyloxycarbonyloxy; 1,3-dioxolane-2-ylmethylene esters, for example 5-methyl-1,3-dioxolane-2-ymetray; C1-6alkoxycarbonylmethyl esters, for example, 1-methoxycarbonylmethylene; aminocarbonylmethyl esters and mono - or di-N-(C1-6alkalemia) options, for example, N,N-dimethylaminocarbonylmethyl esters and N-arylaminovinylketones esters; and they may be formed by any carboxypropyl in the compounds of the present invention. Cleaved in vivo by a complex ester compounds of the invention containing a hydroxy group is, for example, pharmaceutically acceptable ester which is cleaved in the human or animal with the receipt of the original hydroxyl group. Suitable pharmaceutically acceptable esters for the hydroxy-group include C1-6alkanolamine esters, for example, acetylene esters; and antilove esters, in which the phenyl group may be substituted by aminomethyl or N-substituted mono - or di-C1-6alkylamidoamines, for example, 4-aminomethylbenzoic esters and 4-N,N-dime is presented in vivo amides of the compounds of the invention. Examples of such biodegradable in vivo amides include N-C1-6alkylated and N, N-di-(C1-6alkyl)amide, such as N-methyl, N-ethyl, N-propyl, N,N-dimethyl, N-ethyl-N-methyl or N,N-diethylamide.

Another aspect of the present invention provides a method of obtaining compounds of formula (I) or their pharmaceutically acceptable salts or prodrugs of (R1, R2, R3T and ring a have the meanings given for formula (I), unless otherwise stated), involves the following stages:

a) reaction of compounds of formula (II)

in which R3is R3or protected R3, a Rbrepresents hydrogen or a suitable hydroxyamino group, with a compound of formula (III)

in which L represents a shift of the group;

and thereafter if necessary:

i) the conversion of compounds of formula (I) into another compound of formula (I);

ii) removing any protective groups; or

iii) education of its pharmaceutically acceptable salts or prodrugs.

Suitable values for L are, for example, halogen or sulfonyloxy group, for example, chlorine, bromine, methansulfonate or toluene-4-sulfonyloxy.

Conegut be subjected to the reaction together in an inert solvent and base, such as N,N-dimethyl formamide/sodium hydride or dichloromethane/sodium hydroxide or acetonitrile/potassium carbonate or in the presence of a catalyst transfer phase, such as Tetra-n-butylmethacrylate. The reaction is appropriately carried out within 1-6 hours, preferably 1-3 hours, at a temperature of 15-30, preferably 20-25 With obtaining the compounds of formula (I).

The compounds of formula (II) may be commercially available or they can be obtained by modification using well-known methods of obtaining industrially available compounds of formula (II), or they can be obtained in the following ways:

Method (i)

The reaction of the compound of formula (IV):

in which Rbis as defined above, with a compound of formula (V)

in which Rcrepresents C1-4alkyl.

The compounds of formula (IV) and (V) are introduced into the reaction in the Reissert reaction conditions, such as in an inert solvent (such as tetrahydrofuran) in the presence of a base (such as ataxic potassium), in the temperature range of 15-30, preferably 20-25, 10-20 hours, preferably 15-17 hours the compound Obtained is separated and dissolved in alcohol, this is metal (such as 10% Pd/C) and cyclohexene. The mixture is heated at a temperature of 60 to 120 S, preferably 70-90 C, for 15 to 25 hours, preferably 16 to 20 hours, to obtain the compounds of formula (II) in which Rarepresents-CO2WITH1-4alkyl.

Method (ii)

The reaction of the compound of formula (VI):

in which Rbis as defined above, with a compound of formula (VII)

in which Rdrepresents C1-4alkyl.

The compounds of formula (VI) and (VII) are introduced into the reaction in the Fischer as, for example, with an organic acid (such as acetic acid), alcohol (such as ethanol) at a temperature of 60 to 90, preferably 75 to 85 C for 1-5 hours, preferably 1-3 hours the compound Obtained is mixed with a strong acid such as polyphosphoric acid) and heated at 90 to 150 C, preferably 100-120°C., for 0.5 to 4 hours, preferably 0.5 to 2 hours to obtain the compounds of formula (II), in which R2represents hydrogen. Then, if desired, R2may not necessarily be converted into another value of R2defined in formula (I), using methods known in this field, such as the techniques described below.

Method (iii)

Cyclization of the compounds of formula (VIII)

The cyclization can be accomplished using boiling compounds under reflux in an organic solvent such as xylene. The compounds of formula (VIII) in a suitable manner to receive the reaction of the compounds of formula (IX)

in which R1, R2and Rbhave the meanings defined above, with a compound of formula (X)

in which Rahas the values defined above. The reaction is suitably carried out in an organic solvent such as an alcohol, particularly methanol, in the presence of a base such as an alkoxide of an alkali metal, in particular sodium methoxide. Appropriate is moderate temperatures from -30 to 20°C.

Method (iv)

According to another modification of the compounds of formula (II) is obtained by cyclization of the compounds of formula (XI)

in which R1and Rbhave the meanings given above, R7represents alkyl, such as methyl, and R8is carboxyamide group, such as alkyl, in particular methyl.

The cyclization can be carried out in conditions Klingemann by heating a solution of the compound in organic solvent such as toluene, and a suitable acid, such as Pia formula (XII)

in which R1, RbR5and R6have the meanings defined above, with a compound of formula (XIII)

in which R7and R8have the meanings defined above in respect to formula (XI). The compound of formula (XII) can be dissolved in dilute acid, such as 1,5 N. model HC1 in the presence of nitrite such as sodium nitrite, at moderately low temperatures from -30 to 0°C., preferably -5°C.

Then the solution is mixed with a solution of the compounds of formula (XIII) in an organic solvent, such as ethanol, in the presence of a solution of a base such as a hydroxide of an alkali metal, for example, an aqueous solution of sodium hydroxide.

The compounds of formula (III), (IV), (V), (VI), (VII), (VIII), (X), (XII) and (XIII) are known or industrially available, or obtained by methods known in this field, using the standard manipulation of industrially available or known materials.

Rcand Rdsubmit C1-4alkyl. Preferably Rcand Rdrepresent methyl or ethyl.

You should also understand that in some of these reactions may be necessary/desirable to protect any sensitive g is Asti. So, if the reagents include groups such as carboxyl or hydroxyl, it may be desirable to protect the group in some of the above-mentioned reactions.

A suitable protecting group for a hydroxyl group is, for example, acyl group, for example, alcoolica group, such as acetyl, arolina group, for example benzoyl, or arylmethylidene group, for example, benzyl. Conditions unprotect for the above protective groups will need to vary depending on the choice of protective groups. So, for example, acyl group, such as alcoolica or arolina group may be removed, for example, by hydrolysis with a suitable base, such as alkali metal hydroxide, e.g. lithium hydroxide or sodium. Alternatively, allmerica group such as benzyl group may be removed, for example, a hydrogenation catalyst such as palladium on coal.

A suitable protecting group for a carboxyl group is, for example, tarifitsiruyutsya group, for example methyl or ethyl group, which may be removed, for example, by hydrolysis with base, such as sodium hydroxide, or, for example, tert-bucilina group that can be removed, e.g. silna group, that can be removed, for example, a hydrogenation catalyst such as palladium on coal.

Protective groups can be removed at any convenient stage of the synthesis using conventional techniques well known in the field of chemistry.

Some of these intermediate compounds may are new, for example, intermediate compounds of formula (II), and as such they are provided as an additional characteristic of the invention.

When you want pharmaceutically acceptable salt of the compounds of formula (I), it can be obtained, for example, by reaction of the compounds with the appropriate acid (which provides a physiologically acceptable anion) or with the corresponding base (which provides a physiologically acceptable cation, or by any other conventional procedure of education of salt.

In accordance with another aspect of the invention provides a pharmaceutical composition that includes a compound of formula (I) as defined here above, or its pharmaceutically acceptable salt or prodrug, in combination with a pharmaceutically acceptable excipient or carrier.

Compositions of the invention could aqueous or oil suspensions, emulsions, dispersible powders or granules, syrups or elixirs), for topical use (for example, in the form of creams, ointments, gels, aqueous or oily solutions or suspensions), for administration by inhalation (for example, in the form of fine powder or a liquid aerosol), for administration by insufflation (for example, in the form of a fine powder). Or for parenteral administration (for example, in the form of a sterile aqueous or oily solution for intravenous, subcutaneous, intramuscular or intradermal injection or as a suppository for rectal administration).

Compositions of the invention can be obtained by conventional procedures using conventional pharmaceutical excipients, well known in this field. Thus, compositions intended for oral use may contain, for example, one or more coloring, sweetening, flavouring and/or preserving drugs.

Suitable pharmaceutically acceptable excipients to form tablets include, for example, inert solvents, such as lactose, sodium carbonate, calcium phosphate or calcium carbonate, granulating and loosening means, such as corn starch or Allenova acid or talc; preservatives, such as ethyl - or propyl-para-hydroxybenzoate, and antioxidants, such as ascorbic acid. The composition of the tablets can be uncoated or coated or to change their disintegration and the subsequent absorption of the active ingredient within the gastrointestinal tract, or to improve their stability and/or appearance, using conventional tools and procedures that are well known in this field.

Compositions for oral administration can be in the form of hard gelatin capsules in which the active ingredient is mixed with an inert solid diluent, for example calcium carbonate, calcium phosphate or kaolin, or as soft gelatin capsules in which the active ingredient is mixed with water or oil, such as peanut oil, liquid paraffin or olive oil.

Aqueous suspensions generally contain the active ingredient in finely powdered form together with one or more suspendresume agents such as sodium carboxymethylcellulose, methylcellulose, hypromellose, sodium alginate, polyvinylpyrrolidone resin tragakant and resin acacia; dispersing or wetting agents, such as the FL condensation of ethylene oxide with long chain aliphatic alcohols, for example, heptadecafluorooctane, or condensation products of ethylene oxide with partial esters derived from fatty acids and exit, such as polyoxyethylenesorbitan, or condensation products of ethylene oxide with long chain aliphatic alcohols, for example, heptadecafluorooctane, or condensation products of ethylene oxide with partial esters derived from fatty acids and exit, such as polyoxyethylenesorbitan, or condensation products of ethylene oxide with partial esters derived from fatty acids and anhydrides of exit, for example, polyethylenterephthalat. Aqueous suspensions may also contain one or more preservatives (such as ethyl or propyl para-hydroxybenzoate), antioxidants (such as ascorbic acid), coloring agents, flavouring agents and/or sweetening agents (such as sucrose, saccharin or aspartame).

Oil suspensions can be prepared by suspendirovanie active ingredient in a vegetable oil (such as peanut butter, olive oil, sesame oil or coconut oil) or mineral oil such as liquid paraffin). Oil suspensions can also contain Genty, above, and flavoring agents may be added to provide a palatable preparation. These compositions may be preserved by the addition of an antioxidant such as ascorbic acid.

Dispersible powders and granules suitable for obtaining aqueous suspension by the addition of water, usually contain the active ingredient together with a dispersing or wetting agent, suspenders agent and one or more preservatives. Examples of suitable dispersing or wetting agents mentioned above. Can also be additional fillers such as sweetening, flavouring and colouring agents.

The pharmaceutical compositions of the invention may also be in the form of emulsions of oil-in-water. The oil phase may represent vegetable oil, such as olive oil or peanut oil, or mineral oil, such as, for example, liquid paraffin, or a mixture of any of them. Suitable emulsifying agents may be, for example, naturally occurring resins such as the resin of acacia or resin tragakant, naturally occurring phosphatides, such as soybean lecithin, esters or partial esters derived from the x esters with ethylene oxide, such as polyoxyethylene sorbitan monooleate. The emulsions may also contain sweetening, flavouring and preservative agents.

Syrups and elixirs may be formulated with sweetening agents such as glycerin, propylene glycol, sorbitol, aspartame or sucrose and can also contain an emollient, a preservative, flavouring and/or colouring agents.

Pharmaceutical compositions may also be in the form of a sterile aqueous or oily suspension for injection, which can be prepared according to known procedures using one or more appropriate dispersing or wetting agents and suspendida agents which have been mentioned above. A sterile preparation for injection may also be a sterile solution or suspension for injection in a non-toxic acceptable for parenteral use diluent or solvent, for example, a solution in 1,3-butanediol.

The suppository composition can be obtained by mixing the active ingredient with a suitable non-irritating excipient which is solid at ordinary temperatures but liquid at rectal temperature and therefore the kao and glycols.

Compositions for topical application such as creams, ointments, gels, and aqueous or oil solutions or suspensions can be usually obtained by combining the active ingredient with ordinary, acceptable for local use carrier or diluent using conventional procedures well known in this field.

Compositions for administration by insufflation may be in the form of a fine powder containing particles with an average diameter of, for example, 30 μm or much less, and the powder or contains one active ingredient, or diluted with one or more physiologically acceptable carriers such as lactose. Then powder for injection for facilities placed in a capsule containing, for example, from 1 to 50 mg of active ingredient for use with the device of turbinella, such as the device used for the injection of a known drug sodium cromoglycate.

Compositions for administration by inhalation may be in the form of a conventional pressurized aerosol adapted to the filing of the active ingredient or in the form of an aerosol containing finely powdered solid, or droplets of liquid substance. Can be used in a conventional device can be adjusted to supply a metered amount of the active ingredient.

For more information on prepared forms of the compositions reader can get in Chapter 25.2 volume 5 of Comprehensive Medicinal Chemistry (Corwin Hansch; Chairman of Editorial Board), Pergamon Press, 1990.

The amount of active ingredient that is combined with one or more fillers to obtain a single dosage form, it is necessary to vary depending on the exposed treatment of the subject and the specific method of administration. For example, a composition intended for oral administration to humans, usually contains, for example, from 0.5 mg to 2 g of active agent, combined with an appropriate and convenient amount of excipients or fillers, which can vary from about 5 to 98 wt.% the whole composition. Dosage forms typically contain from 1 mg to about 500 mg of the active ingredient. For additional information about the ways and schemes dosage reader can refer to Chapter 25.3 volume 5 of Comprehensive Medicinal Chemistry (Corwin Hansch; Chairman of Editorial Board), Pergamon Press, 1990.

The size of the dose of the compounds of formula I for therapeutic or prophylactic purposes will, of course, vary in accordance with the nature and severity of disease, age and sex of the animal or patient and injected into the soo is possible diseases or medical conditions, which are wholly or partly a consequence of the effect of farnesiana rats.

When using the compounds of formula I for therapeutic or prophylactic purposes it usually should be introduced so that the daily dose was in the range of, for example, from 0.5 mg to 75 mg/kg of body weight, if necessary, introduce fractional doses. When using parenteral route usually given lower doses. For example, for intravenous administration is usually applied dose in the range, for example, from 0.5 mg to 30 mg/kg of body weight. Similarly, for administration by inhalation is applied dose in the range, for example, from 0.5 mg to 25 mg/kg of body weight. However, preferably oral administration.

In accordance with another aspect of the present invention provides a compound of formula (I) or its pharmaceutically acceptable salt or prodrug, as defined here above, for use in a method of therapeutic treatment of the human or animal. As described above, the invention provides a suitable method for the treatment of inflammatory diseases of the introduction of the compounds of formula (I) or its pharmaceutically acceptable salt, prodrug or pharmaceutical compositions.

I salt or a prodrug for use as a medicine.

This compound of formula (I) or its pharmaceutically acceptable salt or prodrug intended for use as a drug for antagonism of the effect mediated MCP-1 in warm-blooded animal such as man.

Thus, in accordance with another aspect of the invention provides the use of compounds of formula (I) or its pharmaceutically acceptable salt or prodrug in the manufacture of a medicinal product for the manifestation of the antagonism of the effect mediated MCP-1 in warm-blooded animal such as man.

In accordance with another aspect of the invention provides a method of providing antagonism of the effect mediated MCP-1 in warm-blooded animal, such as man, in need of such treatment, which includes the introduction of specified animal an effective amount of the compounds of formula (I) or its pharmaceutically acceptable salt or prodrug, as defined here above.

Biological tests

The following methods of biological testing, data and examples serve to illustrate the present invention.

Abbreviations:

ATS American Type Culture Collection, Rockville, USA (Ammeyah protein analysis)

BSA Bovine serum albumin

DMEM Medium Needle, modified by Dulbecco

EGTA Ethylenebis(oxyethylenenitrilo)tetraoxane acid

FCS Fetal calf serum

HEPES (N-[2-Hydroxyethyl]piperazine-N'-[2-econsultancy acid])

HBSS Balanced salt solution Hanks

hMCP-1 Human monocytic chemoattractant protein-1

PBS Saline solution with phosphate buffer

PCR Polymerase chain reaction

As a source of heat-resistant DNA polymerase using AMPLITAQ supplied by the company Perkin-Elmer Cetus.

Binding buffer is 50 mm HEPES, 1 mm CaCl2, 5 mm MgCl2and 0.5% fetal calf serum, brought to a pH of 7.2 using 1 M NaOH.

Nonessential amino acids (100X concentrate) are L-alanine, 890 mg/l; L-asparagine, 1320 mg/l; L-aspartic acid, 1330 mg/l; L-glutamic acid, 1470 mg/l; glycine, 750 mg/l; L-Proline, 1150 mg/l, and L-serine, 1050 mg/l

Additive gipoksantina and thymidine (50x concentrate) is a gipoksantin, 680 mg/l, and thymidine, 194 mg/l

Penicillin-streptomycin is: Penicillin G (sodium salt); 5000 IU/ml; streptomycinresistant 5000 µg/ml.

Synthetic environment for cell culture RPMI 1640 was obtained from Gibco; it contains inorganic salts [CA(NO3)24H2O 100 mg/l; KCl 400 mg/l; MgSO47H2O 100 mg/l; NaCl 6000 mg/l; NaHCO32000 mg/l and Na2HPO4(anhydride) 800 mg/l], D-glucose 2000 mg/l, restored glutathione 1 mg/l, amino acids and vitamins.

FURA-2/AM is pentastomiasis ester 1-[2-(5-carboxykinase-2-yl)-6-aminobenzene-5-oxy]-2-(2'-amino-5'-methylphenoxy)-ethane-N,N,N',N'-tetraoxane acid and was obtained from Molecular Probes, Eugene, Oregon, USA.

The buffer for the deposition of blood contains 8.5 g/l NaCl and 10 g/l hydroxyethyl cellulose.

Buffer for lysis is 0.15 M NH4Cl-, 10 mm knso3, 1 mm EDTA (ethylenediaminetetraacetic acid).

Binding buffer whole cell is 50 mm HEPES, 1 mm CaCl2, 5 mm MgCl2and 0.5% BSA, 0.01% NaN3, brought to pH of 7.2 1M NaOH.

Buffer for washing is 50 mm HEPES, 1 mm CaCl2, 5 mm MgCl2and 0.5% FCS, heat inactivated, 0.5 M NaCl, pH increased to 7.2 1M NaOH.

You can follow the General procedures of molecular biology by any of the methods described in "Molecular Cloning - A Laboratory Manual" Seco>The DNA receptor In MCP-1 (CCR2B) clone using PCR from RNA of cells TNR-1 using suitable oligonucleotide primers based on the published sequence data of the receptor for MCP-1 (Charo et al., 1994, Proc. Natl. Acad. Sci. USA, 91, 2752). The resulting PCR products clone into the vector PCR II (InVitrogen, San Diego, CA). Error-free CCR2B cDNA subcloning as fragment Hind III-Not I in the eukaryotic expression vector pCDNA3 (InVitrogen) to obtain, respectively, pCDNA3/CC-CKR2A and pCDNA3/CCR2B.

Linearized DNA pCDNA3/CCR2B transfection cells Cho-K1 by precipitation of calcium phosphate (Wigler et al., 1979, Cell 16, 777). Transfection cells are harvested by adding genericinterface (G418, Gibco BRL) at a concentration of 1 mg/ml 24 h after transfection cells. Obtaining RNA and Northern blotting carried out as described previously (Needham et al., 1995, Prot.Express.Purific., 6, 134). Clone 7 CHO-K1 (CHO-CCR2B) identify as the most high expressor receptor In MCP-1.

ii) Obtaining fragments of membranes

Cells CHO-CCR2B grown in DMEM with addition of 10% fetal calf serum, 2 mm glutamine, 1x nineteenth amino acids, 1x gipoksantina and timeinfo Supplement, and penicillin-streptomycin (at a concentration of 50 μg streptomycin/ml, Gibco BRL). Fragments of membranes polucha, J. Biol.Chem., 265, 19658). The concentration of the protein is assessed through analysis of protein ICA (Pierce, Rockford, Illinois) according to the manufacturer's instructions.

iii) Analysis

125I MCP-1 is obtained using conjugation of Bolton and Hunter (Bolton et al., 1973, Biochem. J., 133, 529; Amersham International plc.]. The analysis of the equilibrium binding is conducted using the method of Ernst et al., 1994, J. Immunol., 152, 3541. In short, add a variable number of MCP-1, labeled125I to 7 µg of purified membranes of cells CHO-CCR2B in 100 ál of binding buffer. After 1 h incubation at room temperature the reaction mixture bind filtered and washed 5 times with a wash tablet (Brandel MLR-96T Cell Harvester) using cold binding buffer. Gaskets filters (Brandel GF/B) before using pre-soaked for 60 min 0.3% polyethylenimine. After filtration of the individual filters in separate test tubes with a capacity of 3.5 ml (Sarstedt No. 55484) and determine the associated MCP-1, labeled125I (LKB 1277 Gammamaster). Research cold competition conducted as described above using 100 mcmol MCP-1, labeled125I, in the presence of various concentrations of unlabeled MCP-1. Nonspecific binding is determined by the inclusion of a 200-fold molar excess of it is etoc CHO-CCR2B, showed that the CCR2B receptor is present in a concentration of 0.2 pcmall/mg protein membrane and selectively and with high affinity binds MCP-1 (IC50=110 pcmall, Kd=120 mcmol). Binding to these membranes are fully reversible and reaches equilibrium after 45 min at room temperature, and a linear relationship between the binding of MCP-1 and the concentration of the membranes of CHO-CCR2B when using MCP-1 in a concentration of from 100 mcmol 500 pcmall.

Compound dissolved in DMSO (5 ml), was tested by competition with 100 mcmol labeled MCP-1 in the range of concentrations (0.01-50 µmol) using vosmerochnyj curves dose-response and calculated concentration IC50.

Tested compounds of the present invention have values IC5050 µm or less described here the analysis of the binding of the receptor hMCP-1.

b) Mediated MCP-1 current calcium in cells TNR-1

Cell line human monocytes TNR-1 grown in synthetic medium for cell culture RPMI 1640 with the addition of 10% fetal calf serum, 6 mm glutamine and penicillin-streptomycin (at a concentration of streptomycin 50 μg/ml, Gibco BRL). Cells TNR-1 washed in HBSS (without CA2+6cells/ml cell Suspension TNR-1 (0.9 ml) is added to a disposable cuvette capacity of 5 ml, containing a magnetic stirrer in the form of sticks and 2.1 ml of pre-warmed (37 C) HBSS containing 1 mg/ml BSA, 1 mm MgCl2and 2 mm CaCl2. The cuvette was placed in a fluorescence spectrophotometer (Perkin Elmer, Norwalk, CT) and pre-incubated for 4 min at 37 C under stirring. The fluorescence recorded during the 70 s and cells are stimulated by addition to the cell 10 with hMCP-1. [CA2+]i measured excitation alternately at 340 nm and 380 nm and then measuring the intensity of fluorescent emission at 510 nm. I hope and serves to display the ratio of the intensity of emitted fluorescence emission after excitation at 340 nm and 380 nm, (R), and assess cytoplasmic [Ca2+] in accordance with the equation:

where Kdfor FURA-2 CA2+at 37°C is taken for 224 nm. Rmaxrepresents the maximum value of the fluorescence determined after addition of 10 mm ionomycin, Rminrepresents the minimum value defined by the subsequent addition of free CA2+solution, STERI excitation 380 nm, defined, respectively, at Rmaxand Rmin.

Stimulation of cells TNR-1 hMCP causes a rapid, transient rise of [CA2+] i in a specific way and depending on the dose. Curves dose-response show the approximate size EU502 nm. Test compounds dissolved in DMSO (10 μl), quantitatively examined for the inhibition of calcium release by adding them to the cell suspension for 10 s before the addition of the ligand and the dimension reduction of the transient rise of [Ca2+li. Compound is also tested for the absence of agonistic activity by adding instead of hMCP-1.

(C) Chemotaxis mediated hMCP-1 and RANTES

Quantitative analysis of chemotaxis in vitro, make use of a cell line of human monocytes TNR-1. The migration of cells through the polycarbonate membrane is measured by counting the passing cells or directly by using the Coulter counter, or indirectly using calorimetric analysis of viability by measuring the cleavage of salt tetrazole mitochondrial respiratory chain (Scudiero, D. A., et al., 1988, Cancer Res., 48, 4827-4833).

Chemoattractant contribute in 96-well microtitration Board the oval membrane without polyvinylpyrrolidone (PVP) with a pore size of 5 μm (NeuroProbe MB series Cabin John, MD 20818, USA) according to manufacturer's instructions. The chemoattractant appropriately diluted in synthetic medium for cell culture RPMI 1640 (Gibco) or with the addition of 2 mm glutamine and 0.5% BSA, or alternative HBSS with CA2+and mg2+without Phenol Red (Gibco) plus 0.1% of BSA. Each dilution Tegaserod in vacuum for 30 min and placed (400 μl) at the bottom of the cell, the cells TNR-1 (5 105100 μl of RPMI 1640+0,5% BSA) and incubated in each cell the top of the camera. For inhibition of chemotaxis the chemoattractant is kept at a constant submaximal concentrations determined previously (1 nm MCP-1) and add at the bottom of the cell together with the test compounds dissolved in DMSO (the final concentration of DMSO <0,05% vol./about.) at various concentrations. The camera incubated for 2 h at 37 C in 5% CO2. The medium is removed from the upper hole, then wash with 200 μl of physiological saline before opening the camera, wiping dry the surface of the membrane and centrifugation 96-well tablets at 600 g for 5 min to collect the cells. The supernatant (150 μl) aspirinum and back into the cells, add 10 μl of cell proliferation reagent, WST-1, {4-[3-(4-itfeel)-2-(4-nitrophenyl)-2H-5-tetrazolyl]-1,3-is an increase of 3 h and the spectral absorption ability of soluble formisano product read on the reader microtitration the plate at 450 nm. Data is introduced into the electronic dynamic table adjusted for any random migration in the absence of chemoattractant and calculate the average absorbance, standard error of the mean and criteria of statistical significance of differences. hMCP-1 causes dependent on the concentration of the migration of cells with a characteristic biphasic response, maximum at 0.5-1.0 nm.

In an alternative form of the above quantitative analysis can be used labeled with fluorescent-labeled cells in order to facilitate the final detection. In this case the used cells TNR-1 have been labelled with a fluorescent label incubation in the presence of 5 mm Calcein AM (glycine complex N,N'-[[3',6'-bis(atomic charges)-3-oxaspiro[isobenzofuran-1(3H),9'-[N]Cantal]-2',1'-diyl]bis(methylene)]bis[N-[2-[(atomic charges)methoxy] -2-oxoethyl]] -bis [(atomic charges) methyl] ester; Molecular Probes) for 45 min in the dark. Cells are harvested by centrifugation and resuspended in HBSS (without phenol red) and CA2+, Mg2+and 0.1% BSA. 50 μl (2 105 cells) of cell suspension was placed on a filter over each cell and, as mentioned above, the block incubated at 37 C for 2 h in an atmosphere of 5% CO2. At the end of incubation, the cells wash away what about the cells, involved or to the bottom side of the filter, or in the bottom hole, appreciate reading values of fluorescence upon excitation at 485 nm, when the wavelength of the emitted radiation 538 nm (fmax, Molecular Devices). Data is introduced into the electronic dynamic table adjusted for any random migration in the absence of chemoattractant and calculate the mean values of the fluorescence, the standard error of the mean percentage of inhibition and IC50the tested compounds, and it is possible to calculate the criteria of statistical significance of differences. In addition to chemotaxis induced MCP-1, this alternative form of analysis used for the measurement of inhibition of chemotaxis induced RANTES (2 nm).

d) Binding to mononuclear cells of peripheral blood (PBMCs)

i) Obtaining human PBMCs

Fresh human blood (200 ml) obtained from donor volunteers, collect natriciteres anticoagulant to obtain the final concentration of 0.38%. Blood mixed with buffer deposition and incubated at 37 C for 20 minutes the Supernatant is collected and centrifuged at 1700 rpm for 5 min (Sorvall RT6000D). The precipitate after centrifugation resuspended 20 m Tubes rotating at 1700 rpm for 30 min (Sorvall RT6000D) and the resulting layer of cells is removed and transferred into 50 ml Falcon tubes. Cells are washed twice in lysis buffer to remove any remaining red blood cells, followed by two washes in RPMI/BSA. Cells resuspended in 5 ml of binding buffer. The number of cells measured on the Coulter counter and add additional binding buffer to obtain a final concentration of 1.25 107s/ml.

ii) Analysis

[125I]MCP-l is obtained using conjugation of Bolton and Hunter (Bolton et al., 1973, Biochem. J., 133, 529; Amersham International plc]. The analysis of the equilibrium binding is conducted using the method of Ernst et al., 1994, J. Inmnunol., 152, 3541. Briefly, 50 µl of labeled125I MCP-1 (final concentration of 100 μmol) is added to 40 ál (5 of 105cells) of cell suspension in a 96-well plate. Compounds diluted in buffer binding of whole cells from the source solution 10 mm in DMSO, added to final volume of 5 μl to maintain in the analysis of the continuous concentration of DMSO 5%. Total binding is determined in the absence of connection. Nonspecific binding is determined by adding 5 ál of cold MCP-1 to obtain a final concentration in the analysis of 100 nm. The final volume in the cells for analysis is brought to 100 μl of buffer binding of whole cells and tablets hermetically sealed. After incuding wash buffer using a washer tablet (Brandel MLR-96T Cell Harvester). Gaskets filters (Brandel GF/B) before using pre-soaked for 60 min in 0.3% polyethylenimine plus 0.2% BSA. After filtration of the individual filters in separate test tubes with a capacity of 3.5 ml (Sarstedt No. 55484) and determine the associated MCP-1, labeled125I (LKB 1277 Gammamaster).

The activity of the studied compounds was determined by repeated twice analysis using shestiotsechnik curves dose-response and determine the concentration of IC50.

For the tested compounds of the present invention in effective dose was not observed any physiologically unacceptable toxicity.

The invention is further illustrated by, but is not limited to the following examples, in which in the absence of other indications were used following the General procedure.

i) N,N-dimethylformamide (DMF) dried over molecular sieves 4A. Anhydrous tetrahydrofuran (THF) is obtained from vials Aldrich SURESEAL . In the absence of other indications other commercially available reagents are used without additional purification. The organic solvent extracts dried over anhydrous gSO4.

ii) Unless otherwise specified, NMR1H,13C and19F register on pribate DMSO-d6with Me4Si or CCL3F. Chemical shifts are expressed in d (M. D.), and the multiplicity of peaks denoted as follows: s, singlet; d, doublet; DD, doublet of doublets; t, triplet; dt, doublet of triplet; q, Quartet; m, multiplet; Shir. s, broad singlet.

iii) Mass spectra were recorded on a Kratos MS spectrometers 9 VG 12-12 quadrupole, VG 70-250 SE, VG ZAB 2-SE or VG-modified AEI.

iv) For TLC analysis using pre-coated TLC plates Merck (silica gel 60 F254, d=0.25 mm).

v) Flash chromatography carried out on silica (Merck Kieselgel Art.9385).

Example 1

N-(3,4-dichlorobenzyl)-5-hydroxyindole-2-carboxylic acid

Sodium hydroxide (2M, 3 ml) is added to a mixed solution of ethyl-N-(3,4-dichlorobenzyl)-5-hydroxyindole-2-carboxylate (0.1 g) in THF (3 ml) and methanol (1.5 ml). The reaction mixture was stirred at ambient temperature for 4 h, the Reaction mixture was concentrated in vacuo and the residue is dissolved in water (5 ml). The solution is acidified by addition of aqueous hydrochloric acid (2M, 4 ml), precipitating the product in a solid white color. The product is filtered, washed with water and dried in vacuum to obtain specified in the header of the connection is to 2.85 (s, 1H); m/z 334 (M-N+).

The procedure described in the above example, again using the appropriate ethylindole-2-carboxylates. Thus obtained the following compounds.

Example 2

N-[(2,3-dichloride-5-yl)methyl]-5-hydroxyindole-2-carboxylic acid

Yield 36%. NMR (CD3SD3) 5,80 (s, 2H), at 6.84 (DD, 1H), of 6.96 (d, 1H), 7,14 (s, 1H), 7.23 percent (d, 1H), 7,73 (d, 1H), of 8.06 (d, 1H); m/z 339 (M-H+) 337, 335.

Example 3

N-(3-chloro-4-terbisil)-5-hydroxyindole-2-carboxylic acid

Yield 68%. NMR (CD3SD3) of 5.75 (s, 2H), PC 6.82 (DD, 1H), 6,95 (m, 2H), 7,12 (s, 1H), 7,2-7,4 (m, 3H); m/z 320 (M-N+), 318.

Example 4

N-(4-chloro-3-terbisil)-5-hydroxyindole-2-carboxylic acid

Yield 94%. NMR (CD3SD3) 5,78 (s, 2H), 6,78 (DD, 1H), 6,80 (DD, 1H), of 6.96 (d, 1H), 7,03 (DD, 1H), 7,12 (s, 1H), 7,31 (d, 1H), 7,43 (t, 1H); m/z 318 (M-N+).

Example 5

N-(3-Chlorobenzyl)-5-hydroxyindole-2-carboxylic acid

A yield of 75%. m/z 300 (M-N+).

Example 6

N-(3-trifloromethyl)-5-hydroxyindole-2-carboxylic acid

Yield 81%. m/z 334 (M-N+).

Example 7

N-(4-Chlorobenzyl)-5-hydroxyindole-2-carboxylic acid

Wichata

Yield 95%. m/z 414 (M-H+).

Example 9

4-Bromo-N-(3,4-dichlorobenzyl)-5-hydroxyindole-2-carboxylic acid

Yield 96%. NMR (CD3SD3) 5,78 (s, 2H), 6,86 (DD, 1H), 7,01 (d, 1H),? 7.04 baby mortality (s, 1H), 7,33 (s, 1H), 7,40 (d, 1H), 7,52 (d, 1H), 9,78 (s, 1H), 13,10 (Shir. s, 1H); m/z 414 (M-H+).

Example 10

N-(3,4-dichlorobenzyl)-5-hydroxy-3-methylindol-2-carboxylic acid

The yield was 73%. NMR (CD3SD3) 2,44 (s, MN), 5,69 (s, 2H), 6,83 (m, 2H), 6,92 (d, 1H), 7,25 (d, 1H), 7,30 (d, 1H), 7,50 (d, 1H), 9,00 (s, 1H), 12,90 (Shir. s, 1H); m/z 350 (M-N+).

Example 11

N-(3,4-dichlorobenzyl)-4-fluoro-5-hydroxyindole-2-carboxylic acid

Yield 68%. NMR (CD3SD3) 5,80 (s, 2H), to 6.88 (m, 1H), 7,00 (t, 1H), 7,20 (m, 2H), 7,32 (m, 1H), 9,25 (s, 1H), 13,10 (s, 1H); m/z (M-N+) 351,9.

Example 12

N-(3,4-dichlorobenzyl)-3-methoxy-5-hydroxyindole-2-carboxylic acid

The yield was 73%. NMR (CD3SD3) 4,3 (s, 3H), 5,7 (s, 2H), 6,9 (m, 2H), and 7.1 to 7.4 (m, 4H); m/z 364, 366 (M-H+).

Example 13

N-(3,4-dichlorobenzyl)-3-chloro-5-hydroxyindole-2-carboxylic acid

Yield 97%. NMR (CD3SD3) of 5.75 (s, 2H), 6,9 (m, 3H), and 7.3 (s, 1H), 7,45 (d, 1H), 7.5 (d, 1H), 9,35 (s, 1H); m/z 368 (M-N+).

Example 14

N-(3,4-dichlorobenzyl)-4-chloro-5-hydroxyindole-2-CA (d, 1H), 7,51 (d, 1H), 9,67 (Shir. s, 1H); m/z 368,2 (M-N+).

Obtaining raw materials

The source materials for the above examples represent or commercially available materials, or easily obtained by standard methods from known materials. For example, the following reactions (Methods A-J) are illustrations and not limitations of obtaining the source materials used in the above reactions.

Method AND

3-Chloro-4-florantyrone

A solution of 3-Chloro-4-forventelige (3 g) in THF (40 ml) is added over 2 min to a stirred suspension of sodium borohydride (1.07 g) in methanol (40 ml) at 0 C. the Mixture is allowed the opportunity to be warmed to room temperature and then the reaction quenched with water. The resulting suspension is divided between water and simple diethyl ether and the combined organic extracts dried and concentrated in vacuo. The residue is dissolved in dichloromethane (90 ml) and added triphenylphosphine (4,62 g) and tetrabromomethane (6,64 g) at 0 C. the Mixture is allowed the opportunity to be warmed to room temperature overnight, then concentrated in vacuo and the residue purified column chromatography using as eluent ISO-hexane to obtain wish is zaldehyde, get:

3-Fluoro-4-Chlorobenzilate

Yield 74%. NMR: 4,5 (s, 2H), and 7.1 (t, 1H), 7,25 (m, 1H), 7,45 (DD, 1H).

Method IN

2,3-Dichloro-5-(hydroxymethyl)pyridine

The complex of borane-tetrahydrofuran (1M solution in tetrahydrofuran, 52 ml) is added to a stirred solution of 5,6-dichloronicotinic acid (2 g) in tetrahydrofuran (60 ml) for 20 min at 0 C. the Reaction mixture is allowed the opportunity to be warmed to room temperature over 90 min and then cooled to C and the reaction quenched with water (100 ml). The solution is saturated with sodium chloride in the form of solids and extracted with ethyl acetate and the combined organic extracts dried and concentrated in vacuo. The residue is ground to powder with dichloromethane-50% ethyl acetate and the solid by-product is removed by filtration. The filtrate was concentrated in vacuo and purified column chromatography using as eluent isohexane/ethyl acetate (1:1 vol./about.) obtaining the product as a white solid (820 mg, 45%). NMR: 4,55 (d, 2H), and 5.5 (t, 1H), 8.0 a (m, 1H), 8,3 (m, 1H); m/z 178,1 (M+N+).

The way TO

2,3-Dichloro-5-(methyl bromide)pyridine

2,3-Dichloro-5-(hydroxymethyl)pyridine (275 mg) was dissolved in dichloromethane (10 ml) and stirred the e and the residue is purified column chromatography using as an eluent of isohexane: 2.5% ethyl acetate to obtain the product as white solid (270 mg, 73%). NMR: the 4.75 (s, 2H), 8,25 (m, 1H), 8,5 (m, 1H); m/z 242 (M+N+).

Method D

Ethyl-5-acetoxy-M-(3,4-dichlorobenzyl)indole-2-carboxylate

(i) Ethyl-5-hydroxyindole-2-carboxylate

Tribromide boron (64,58 g) is added dropwise to a stirred solution of ethyl-5-methoxyindol-2-carboxylate (20 g) in dichloromethane (1000 ml) at -78 C in argon atmosphere. The reaction mixture is allowed the opportunity to be warmed to room temperature and stirred for another 2 h, the Reaction mixture was poured into a solution of a mixture of ice/saturated aqueous sodium hydrogen carbonate solution under stirring and extracted with ethyl acetate. The combined organic extracts washed with saturated aqueous sodium hydrogen carbonate solution, water, aqueous saturated solution of sodium chloride and dried. The solution was concentrated in vacuo and the residue purified column chromatography using as eluent a mixture of 0-60% diethyl ether: ISO-hexane to obtain the product as a white solid (9,02 g, 48%). NMR: is 1.31 (t, 3H), 4,29 (kV, 3H), 6,79 (DD, 1H), 6.90 to (DD, 1H), 7,22 (d, 1H), 8,84 (s, 1H), to 11.52 (Shir.s, 1H); m/z 206 (M+N+).

ii) Ethyl-5-acetoxymethyl-2-carboxylate

Mix a solution of ethyl-5-hydroxyindole-2-carboxylate the offer was concentrated in vacuo and the residue is dissolved in acetylacetone. The combined organic extracts washed with hydrochloric acid (2M), saturated aqueous sodium hydrogen carbonate solution, water, aqueous saturated solution of sodium chloride and dried. The solution was concentrated in vacuo to obtain the product in a solid yellow color (9,39 g, 100%). NMR: of 1.20 (t, 3H), 2,10 (s, 3H), 4,19 (kV, 2H), 6,86 (DD, 1H), 6,97 (d, 1H), 7,20 (s, 1H), 7,29 (d, 1H); m/z 248 (M+N+).

iii) Ethyl-5-acetoxy-N-(3,4-dichlorobenzyl)indole-2-carboxylate

3,4-dichlorobenzamide (5,96 g) added to a mixed solution of ethyl-5-acetoxymethyl-2-carboxylate (5.4 g) and potassium carbonate (6,94 g) in acetonitrile (500 ml) in an argon atmosphere. The reaction mixture is heated at 80 C for 16 h, then concentrated in vacuo and the residue distributed between ethyl acetate and water. The combined organic extracts washed with water, saturated aqueous sodium chloride and dried. The solvent is removed in vacuum and the residue is ground to powder with isohexane with getting the product in the form of a cream solid (5,55 g, 63%). NMR: of 1.27 (t, 3H), and 2.27 (s, 3H), 4,28 (kV, 2H), of 5.82 (s, 2H), 6.90 to (d, 1H), to 7.09 (DD, 1H), 7,33-7,40 (m, 2H), 7,46 (d, 1H), 7,52 (d, 1H), 7,60 (d, 1H).

The procedure described in method D (i)-(iii), again using sootvetstvujushej the corresponding benzylalkonium. Thus obtained the following compounds.

Method D1

Ethyl-5-acetoxy-N-[(2,3-dichloride-5-yl)methyl]indole-2-carboxylate

Output 90%. NMR: of 1.27 (t, 3H), and 2.26 (s, 3H), 4,28 (kV, 2H), to 5.85 (s, 2H), 7,12 (DD, 1H), 7,38 (s, 1H), 7,47 (d, 1H), 7,68 (d, 1H), 7,78 (d, 1H), 8,10 (d, 1H); m/z 409 (M+N+), 407.

Method D2

Ethyl-5-acetoxy-N-(3-chloro-4-terbisil)indole-2-carboxylate

Yield 57%. NMR (Dl3): of 1.37 (t, 3H), of 2.33 (s, 3H), 4,35 (kV, 2H), 5,74 (s, 2H), 6.90 to (m, 1H), 7,00 (d, 1H), 7,05 (DD, 1H), 7,13 (DD, 1H), 7,26 (s, 1H), was 7.36 (s, 1H), 7,22 (d, 1H).

Ethyl-5-acetoxy-N-(4-chloro-3-terbisil)indole-2-carboxylate

The yield was 73%. m/z 390 (MH+).

Ethyl-5-acetoxy-N-(3-Chlorobenzyl)indole-2-carboxylate

Yield 93%. m/z 372 (MN+).

Ethyl-5-acetoxy-N-(3-trifloromethyl)indole-2-carboxylate

Yield 91%. m/z 406 (MN+).

Ethyl-5-acetoxy-N-(4-Chlorobenzyl)indole-2-carboxylate

Yield 70%. m/z 372 (MN+).

Ethyl-3-acetoxy-3-bromo-N-(3,4-dichlorobenzyl)indole-2-carboxylate

Yield 86%. m/z 486 (MH+).

Ethyl-5-acetoxy-4-bromo-N-(3,4-dichlorobenzyl)indole-2-carboxylate

Yield 62%. NMR: of 1.40 (t, 3H), 2,39 (s, 3H), of 4.38 (q, 2H), 5,77 (s, 2H), PC 6.82 (DD, 1H), was 7.08 (d, 1H), 7,18 (s, 1H), 7,22 (d, 1H), 7,32 (d, 1H), 7,42 (s, 1H); m/z 486 (MT, 3H), a 2.36 (s, 3H), 2.40 a (s, 3H), 4,35 (kV, 2H), USD 5.76 (s, 2H), 6,83 (DD, 1H), 7,00 (d, 1H), 7,10 (d, 1H), 7,19 (s, 1H), 7,30 (d, 1H), 7,40 (s, 1H); m/z 421 (MH+).

Ethyl-5-acetoxy-N-(3,4-dichlorobenzyl)-3-Clorinda-2-carboxylate

Yield 83%. NMR: a 1.25 (t, 3H), of 2.25 (s, 3H), 4,3 (kV, 2H), and 5.8 (s, 2H), 6,9 (d, 1H), 7,2 (m, 1H), and 7.4 (m, 2H), 7.5 (d, 1H), 7.7 (d, 1H); m/z 441,8 (M+N+).

Method E

Ethyl-N-(3,4-dichlorobenzyl)-5-hydroxyindole-2-carboxylate

Ethoxide sodium (1.86 g) is added to a mixed solution of ethyl-5-acetoxy-N-(3,4-dichlorobenzyl)indole-2-carboxylate (of 5.55 g) in ethanol (50 ml) in an argon atmosphere. The reaction mixture was stirred at room temperature for 2 h, then concentrated in vacuo and the residue acidified with aqueous hydrochloric acid (2M) and extracted with dichloromethane. The combined organic extracts washed with water, saturated aqueous sodium chloride and dried. The solvent is removed in vacuum and the residue is ground to powder with hexane/diethyl ether to obtain the product as a white solid (3,17 g, 92%). NMR: 1.26 in (t, 3H), 4,25 (kV, 2H), of 5.75 (s, 2H), for 6.81-6,91 (m, 2H), 6,98 (d, 1H), 7,19 (s, 1H), 7,29 (d, 1H), 7,38 (d, 1H), 7,50 (d, 1H), 9,06 (s, 1H); m/z 364 (M+N+).

Method F

Ethyl-5-acetoxy-3-bromoindole-2-carboxylate

N-br the Reaction mixture is stirred for 4 h, then pour in the water. The precipitate is filtered and dried in vacuum to obtain specified in the title compound as a white powder (0,23 g, 87%). NMR: to 1.38 (t, 3H), of 2.23 (s, 3H), of 4.38 (kV 2H), 7,10 (DD, 1H), 7.23 percent (d, 1H), 7,50 (d, 1H), to 12.28 (Shir.s, 1H); m/z 326 (M+).

The way F1

Ethyl-5-acetoxy-3-Clorinda-2-carboxylate

A solution of ethyl-5-acetoxymethyl-2-carboxylate (500 mg) in dichloromethane (10 ml) was stirred at room temperature in the presence of N-chlorosuccinimide (297 mg) and potassium carbonate (279 mg) during the night. The precipitate is collected by filtration, washed with cold dichloromethane followed by washing with water and dried in vacuum overnight to give the desired product as a white powder (425 mg, 75%). NMR: of 1.35 (t, 3H), of 2.25 (s, 3H), 4,4 (kV, 2H), and 7.1 (d, 1H), and 7.3 (s, 1H), 7.5 (d, 1H), and 12.2 (s, 1H); m/z 281,9 (M+N+).

Method G

Ethyl-5-acetoxy-3-methylindol-2-carboxylate

(i) Ethyl-5-methoxy-3-methylindol-2-carboxylate

Concentrated sulfuric acid (1 ml) are added to a solution of the hydrochloride of 4-methoxyphenylhydrazine (11.2 g) and 2-hydroxybutyric acid (8,72 g) in ethanol (250 ml) and the solution heated to boiling under reflux for 16 hours, the Reaction mixture is cooled, concentrated th substances (8.8 g, 59%). NMR: of 1.36 (t, 3H), 3,76 (s, 3H), 4,30 (kV, 2H), to 6.88 (DD, 1H), 7,03 (d, 1H), 7,28 (d, 1H), 11,28 (Shir.s, 1H); m/z 232 (M-N+).

ii) Ethyl-5-acetoxy-3-methylindol-2-carboxylate

Tribromide boron (25 g) is added dropwise to a stirred solution of ethyl-5-methoxy-3-methylindol-2-carboxylate (2.0 g) in dry dichloromethane (250 ml) at -78 C in argon atmosphere. The reaction mixture is allowed to warmed to room temperature and stirred for another 2 h, the Reaction mixture was poured into a solution of ice/saturated aqueous sodium hydrogen carbonate solution under stirring and extracted with ethyl acetate. The combined organic extracts washed with saturated aqueous sodium bicarbonate, water, saturated aqueous sodium chloride and dried (MgSO4). The solution was concentrated in vacuo and the residue is dissolved in ethyl acetate. Add DMAP (dimethylaminopyridine, 20 mg) and acetic anhydride (0.5 ml) and the solution heated to boiling under reflux for 5 minutes, the Reaction mixture was cooled, concentrated in vacuo and the residue triturated with simple ether to obtain specified in the title compound as a white powder (0.4 g, 18%). NMR: of 1.37 (t, 3H), of 2.25 (s, 3H), of 2.50 (s, 3H), 4,34 (kV, 2H), 7,00 (DD, 1H), 7,37 (d, 1H), 7,40 (d, 1H), to 11.52 (Shir.s, 1H); m/z 260 CLASS="ptx2">

Ethyl-5-acetoxy-4-bromoindole-2-carboxylate NMR: of 1.42 (t, 3H), 2,39 (s, 3H), 4,42 (kV, 2H), 7,02 (d, 1H), 7.23 percent (s, 1H), 7,35 (d, 1H), which 9.22 (Shir.s, 1H); m/z 324, 326 (M-N+).

Way N

Methyl-N-(3,4-dichlorobenzyl)-4-fluoro-5-hydroxyindole-2-carboxylate

(i) 2-fluoro-3-benzyloxybenzaldehyde

2-fluoro-3-hydroxybenzaldehyde (16,49 g) dissolved in dimethylformamide (200 ml) and stirred in an argon atmosphere. Add sodium hydride (60% in mineral oil, 5,18 g) and the mixture is stirred for 30 minutes Add benzylbromide (16,8 ml) and the mixture is stirred over night. The reaction mixture was concentrated in vacuo and the resulting residue partitioned between diethyl simple ether (200 ml) and water (200 ml). The combined organic extracts washed with water (400 ml), dried (MgSO4) and concentrated in vacuo. The residue is purified column flash chromatography using a gradient of 0-10% ethyl acetate/isohexane as eluent to obtain the product as yellow solid (18,41 g).1H NMR (DMSO-d6) 5,20 (s, 2H), 7,2-7,6 (m, 8H), of 10.21 (s, 1H).

(ii) Methyl-2-azido-3-(2-fluoro-3-benzyloxyphenyl)propanoic

A mixture of methylacetoacetate (36,64 g) and 2-fluoro-3-benzyloxybenzaldehyde (18,32 g) in methanol (250 ml) was added capl is left stirring for 20 min, give be heated up to 5 and stirred over night.

The precipitate is filtered off, then washed successively with cold methanol, dilute solution of acetic acid in water and water. The obtained solid is dried in vacuum to obtain the product as a pale brown solid (16,70 g), used without purification.

(iii) Methyl-4-fluoro-5-benzyloxyindole-2-carboxylate

A solution of methyl-2-azido-3-(2-fluoro-3-benzyloxyphenyl)propanoate (16.7 g) in xylene (600 ml) is added dropwise with stirring to boiling under reflux xylene (2.4 l) for 1 h, and then stirred for another 20 minutes, the Reaction mixture was concentrated in vacuo and purified column flash chromatography using a gradient of 0-100% ethyl acetate/isohexane as eluent to obtain the product as yellow solid (12,93 g). 1H NMR (DMSO-d6) 3,85 (s, 3H), of 5.15 (s, 2H), 7,05 was 7.45 (m, 8H), 12,06 (s, 1H); m/z (+) 300,4 (MN+).

(iv) Methyl-N-(3, 4-dichlorobenzyl) -4-fluoro-5-benzyloxyindole-2-carboxylate

Sodium hydride (60% in mineral oil, 589 mg) are added to a solution of methyl 4-fluoro-5-benzyloxyindole-2-carboxylate (4 g) in dimethylformamide (100 ml) and the mixture is displaced is their night. The reaction mixture was concentrated in vacuo and the residue partitioned between diethyl ether (100 ml) and water (100 ml). The organic extracts washed with water (100 ml), dried (MgSO4), concentrated in vacuo and purified flash column-chromatography using isohexane followed by washing with 5% ethyl acetate/isohexane as eluent to obtain the product as a yellow crystalline solid (4.61 in).1H NMR (DMSO-d6) of 3.80 (s, 3H), of 5.15 (s, 2H), 5,80 (s, 2H), 6,85 (m, 1H), 7,25-7,52 (m, 10H); m/z (+) 458,2 (MN+).

(v) Methyl-N-(3,4-dichlorobenzyl)-4-fluoro-5-hydroxyindole-2-carboxylate

A mixture of methyl-N-(3,4-dichlorobenzyl)-4-fluoro-5-benzyloxyindole-2-carboxylate (by 8.22 g) and 5% Pd/C (200 mg) in ethyl acetate (250 ml) is stirred under an atmosphere of hydrogen overnight, filtered through celite. Concentrated in vacuo and purified column flash chromatography using a gradient of 0-50% ethyl acetate/isohexane as eluent to obtain the product as a brown solid (6,18 g).1H NMR (DMSO-d6) of 3.80 (s, 3H), of 5.75 (s, 2H), 6,85 (m, 1H), 7,00 (t, 1H), 7,22 (m, 2H), 7,30 (m, 1H), 7,50 (m, 1H); m/z (-) 366,2 (MN-).

The way I

Ethyl-N-(3,4-dichlorobenzyl)-3-methoxy-5-hydroxyindole-2-carboxylate

(i) Heterochiral-2-carboxylate in ethyl acetate (40 ml) and acetic acid (20 ml). The mixture is stirred for 18 h and then distributed between ethyl acetate and water. The organic extracts washed with water, saturated aqueous sodium bicarbonate and dried. The solvent is removed in vacuum and the obtained resin was triturated with diethyl simple ether to obtain the product as an orange powder (1.8 g). NMR: a 1.45 (t, 3H), and 4.5 (q, 2H), 5,1 (s, 2H), 7,05 (m, 2H), and 7.3 (m, 5H), 7,9 (d, 1H); m/z 322 (M+N+).

(ii) Ethyl-3-methoxy-5-benzyloxyindole-2-carboxylate

Bodyoctane (300 mg) is added to a mixed solution of ethyl-5-benzyloxyindole-2-carboxylate (2.0 g) in toluene (100 ml) and methanol (10 ml). The mixture is refluxed in an inert atmosphere for 2.5 hours the Solution was concentrated in vacuo and the residue purified column chromatography using 30-50% diethyl simple ether/isohexane to obtain an orange solid (1,41 g). NMR: 1,4 (t, 3H), of 4.05 (s, 3H), 4,4 (kV, 2H), 5,1 (s, 2H), and 7.1 (DD, 1H), 7,2-7,5 (m, 8H)); m/z 326 (MN+).

(iii) Ethyl-N-(3, 4-dichlorobenzyl) -3-methoxy-5-benzyloxyindole-2-carboxylate

3,4-DICHLOROSILANE (0,72 ml) is added to a mixed solution of ethyl-3-methoxy-5-benzyloxyindole-2-carboxylate (1.40 g), potassium carbonate (0,90 g) and potassium iodide (0.1 g)racedata and water. The combined organic extracts washed with water, then 3 times with saturated aqueous sodium chloride and dried. The solvent is removed in vacuo and the residue purified column chromatography using 10-30% ethyl acetate/isohexane with getting a yellow oil (0.9 g). NMR: 1,4 (t, 3H), 4,0 (s, 3H), 4,4 (kV, 2H), 5,1 (s, 2H), 5,6 (s, 2H), 6,9 (DD, 1H), 7,0-7,5 (m, N); m/z 484 (MH+).

(iv) Ethyl-N-(3,4-dichlorobenzyl)-3-methoxy-5-hydroxyindole-2-carboxylate

5% Pd/C (100 mg) is added to a mixed solution of ethyl-N-(3,4-dichlorobenzyl-3-methoxy-5-benzyloxyindole-2-carboxylate (0.9 g) in ethyl acetate (50 ml) and the mixture hydronaut within 12 hours the Catalyst is filtered off and the filtrate is evaporated to obtain a brown oil (0,61 g), which is used without further purification. NMR: 1,4 (t, 3H), 4,0 (s, 3H), 4,4 (kV, 2H), 5,6 (2N), 6,8 (DD, 1H), 6,9 (DD, 1H), and 7.1 (m, 3H), and 7.3 (d, 1H)); m/z 394 (MH+).

Method J

Ethyl-N-(3,4-dichlorobenzyl)-4-chloro-5-methoxyindol-2-carboxyla

(i) Ethyl-2-azido-3-(2-chloro-3-methoxyphenyl)propenoate

The solution ethylacetoacetate (9,9 g) and 2-chloro-3-methoxybenzaldehyde (3 g) in ethanol (20 ml) is added dropwise into a solution of ethoxide sodium (4.7 g) in ethanol (10 ml) at 0 C. the Reaction mixture is allowed to warmed to the temperature of acusat (gSO4), concentrated in vacuo and the residue purified column chromatography using isohexane-12% ethyl acetate/ isohexane as eluent to obtain the product as a pale yellow crystalline solid (2.2 g, 44%). It is used without further purification.

ii) Ethyl-4-chloro-5-methoxyindol-2-carboxylate

A solution of ethyl-2-azido-3-(2-chloro-3-methoxyphenyl)propenoate (2,22 g) in xylene (100 ml) is heated at the boil under reflux for 30 min, concentrated in vacuo and the residue purified column chromatography using isohexane-50% ethyl acetate as eluent to obtain the product as pale yellow solid (1,34 g, 67%). NMR: (Dl3) is 1.31 (t, 3H), of 3.84 (s, 3H), 4,32 (kV, 2H), 7,0 (d, 1H), 7,22 (d, 1H), 7,39 (d, 1H), 12,2 (user.s, 1H).

(iii) Ethyl-N-(3,4-dichlorobenzyl)-4-chloro-5-methoxyindol-2-carboxyla

Sodium hydride (60 mg) are added to a solution of ethyl-4-chloro-5-methoxyindol-2-carboxylate (250 mg), 3,4-dichlorobenzaldehyde (of 0.21 ml) and tetrabutylammonium (3 mg) in DMF at ambient temperature in an inert atmosphere. The reaction mixture was stirred at ambient temperature for 18 h, and then distributed between ethyl acetate (30 ml) and water (a using isohexane-15% ethyl acetate as eluent to obtain the product as a white solid (196 mg, 48%). NMR: (Dl3) of 1.39 (t, 3H), 3,93 (s, 3H), 4,32 (kV, 2H), 5,73 (s, 2H), at 6.84 (DD, 1H), 7,06-7,16 (m, 3H), 7,31 (d, 1H), 7,42 (s, 1H).

iv) Ethyl-N-(3,4-dichlorobenzyl)-4-chloro-5-hydroxyindole-2-carboxylate

Trimethylsilylmethyl (0.6 ml) are added to a solution of ethyl-N-(3,4-dichlorobenzyl)-4-chloro-5-methoxyindol-2-carboxylate (190 mg) in chloroform (20 ml). The mixture is heated to 50 C for 18 h, then poured into methanol (50 ml) and concentrated in vacuo. The residue is purified column chromatography using a mixture of isohexane-20% ethyl acetate/isohexane as eluent to obtain the product as yellow solid (100 mg, 71%). NMR: (Dl3) of 1.39 (t, 3H), 4,35 (kV, 2H), 5,72 (s, 2H), at 6.84 (DD, 1H), 7,05-7,13 (m, 3H), and 7.3 (d, 1H), 7,35 (s, 1H); m/z 396, 2/398,2 (M-N+).

Example 15

N-(3,4-Dichlorobenzyl)-2-triftormetilfullerenov-5-hydroxyindole

The sodium methoxide (21 mg) is added to a stirred solution of N-(3,4-dichlorobenzyl)-2-triftormetilfullerenov-5-acetoxyethyl (90 mg) in methanol (10 ml). The reaction mixture was stirred at ambient temperature for 1.5 h, then concentrated and acidified by addition of aqueous hydrochloric acid (2M, 5 ml), extracted with dichloromethane and concentrated in vacuo to obtain korichnevom>The source material was obtained as follows:

(i) N-(3, 4-Dichlorobenzyl)-5-acetoxymethyl-2-carboxylic acid

Dimethylaminopyridine (100 mg) and acetic anhydride (1,12 ml) are added to a solution of N-(3,4-dichlorobenzyl)-5-hydroxyindole-2-carboxylic acid in ethyl acetate (50 ml) and stirred at ambient temperature for 1 h, Add ethanol (10 ml) and the reaction mixture is stirred for 30 minutes the Solvent was partially evaporated and add isohexane obtaining a precipitate, which is filtered off and dried to obtain a white solid (1.12 g). NMR: of 2.25 (s, 3H), of 5.85 (s, 2H), 6,9 (DD, 1H), and 7.3 and 7.6 (m, 5H); m/z 376, 378 (M-H+).

(ii) N-(3,4-Dichlorobenzyl)-2-triftormetilfullerenov-5-acetoxyethyl

To a stirred solution of N-(3,4-dichlorobenzyl)-5-acetoxymethyl-2-carboxylic acid in DMF (5 ml) under inert atmosphere added HATU (0.27 g), DIPEA (0,12 ml) and triftormetilfullerenov (97 mg). The reaction mixture was stirred at ambient temperature for 18 hours the Mixture was poured into a saturated solution of sodium bicarbonate and the resulting precipitate filtered and dried to obtain the product (90 mg). NMR: of 2.25 (s, 3H) and 5.9 (s, 2H), 6,9 (DD, 1H), 7,0 (DD, 1H), and 7.1 (s, 1H), 7,35 (m, 1H); m/z 506, 508 (M-N+).

Example 16

+).

The source material was obtained as follows:

Methanesulfonanilide (0.5 ml) is added to a cooled (0 C) solution of N-(3,4-dichlorobenzyl)-5-acetoxymethyl-2-carboxylic acid (1.12 g) in pyridine (30 ml) and stirred at 0 for 1.5 hours Gaseous ammonia bubbled through the reaction mixture for 15 min, then the excess ammonia is removed in vacuum. The reaction mixture is cooled to 0 C and stirred solution add methylsulfonylamino (2.5 ml) and provide an opportunity for 18 h to reach the ambient temperature. Add methanesulfonanilide (2 ml) and reaction is washed 3 times with a mixture of aqueous hydrochloric acid (1M) and saturated solution of ammonium chloride (1:1). The organic extracts are dried, concentrated in vacuo and the residue purified column chromatography using 10-25% ethyl acetate/ISO-hexane to give the desired product (300 mg). NMR: of 2.25 (s, 3H), 5,6 (s, 2H), 7,0 (DD, 1H), 7,45-the 7.65 (m, 4H), 7.7 (d, 1H).

Example 17

N-(3,4-dichlorobenzenesulfonyl)-5-hydroxyindole-2-carboxylic acid

A solution of anhydrous lithium iodide (870 mg) and methyl-N-(3,4-dichlorobenzenesulfonyl)-5-hydroxyindole-2-carboxylate (260 mg) in pyridine (15 ml) is stirred while boiling under reflux for 4 hours the Reaction mixture is cooled and concentrated in vacuo. The residue is dissolved in water (20 ml) and acidified with acetic acid. The product is extracted with ethyl acetate and the combined extracts are dried, concentrated in vacuo and the residue purified column chromatography using as eluent 1% acetic acid containing dichloromethane-50% ethyl acetate to give the desired product as a glass (72 mg, 29%). NMR: 6,9 (m, 2H), 7,25 (s, 1H), a 7.85 (d, 1H), 7,9 (m, 2H), 8,15 (s, 1H), and 9.5 (s, 1H); m/z 385,8 (M-N+).

The source material was obtained as follows:

(i) Methyl N-(3,4-dichlorobenzenesulfonyl)-5-benzyloxyindole-2-carboxylate

Sodium hydride (60% dispersion, 444 mg) to relax the ur. After 1 h, add 3,4-dichlorobenzenesulfonate (2,72 g). Stirring is continued for 2 h, after which the reaction mixture is distributed between water and ethyl acetate. The combined organic extracts dried and concentrated in vacuo, and the residue is purified column chromatography using as eluent a mixture of isohexane-20% ethyl acetate to give the desired product as a white solid (2,02 g, 56%). NMR: 3,85 (s, 3H), of 5.1 (s, 2H), 7,2 (m, 1H), and 7.4 (m, 7H), and 7.9 (s, 2H), and 8.0 (d, 1H), and 8.2 (s, 1H); m/z 489,8 (MN+).

(ii) Methyl N-(3, 4-dichlorobenzenesulfonyl)-5-hydroxyindole-2-carboxylate

A suspension of 5% palladium on coal in ethyl acetate (450 ml) and methyl-N-(3,4-dichlorobenzenesulfonyl)-5-benzyloxyindole-2-carboxylate (2,01 g) was stirred at 60 C in an atmosphere of hydrogen at atmospheric pressure for 48 hours, the Catalyst was removed by filtration and the filtrate concentrated in vacuo. The residue is purified column chromatography using as eluent 20% ethyl acetate/isohexane to give the desired product in the form of resin (270 mg, 16%). NMR: 3,85 (s, 3H), 7,0 (m, 2H), 7,35 (s, 1H), 7,9 (m, 3H), and 8.1 (s, 1H), and 9.6 (s, 1H); m/z 401,9 (MN+).

Example 18

N-(3,4-dichlorobenzyl)-5-acetoxymethyl-2-carboxylic acid

To estamineria (100 mg) and acetic anhydride (5.0 ml) and the resulting mixture is stirred for 2 hours The organic extracts are washed with 1 N. HCl and dried. Add hexane to induce crystallization of the product. The solid is filtered off and washed with hexane to give the desired product (5 g, 44%).1H NMR (DMSO-d6) to 2.25 (s, 3H), of 5.85 (s, 2H), 6,9 (DD, 1H), 7,05 (DD, 1H), and 7.3 and 7.6 (m, 5H); m/z 378, 380 (MN+).

Example 19

Pharmaceutical compositions

This example illustrates, but without limiting their corresponding defined in this application to pharmaceutical dosage forms of the invention (active ingredient referred to as “Compound X”), for therapeutic or prophylactic use in humans:

Example

Compound X in the above compositions may include the connection specified in examples 1-3.

The above composition can be obtained by conventional procedures well known in the pharmaceutical field. The tablets (a)-(C) can be coated with enteric shell using conventional means, for example, ensuring coverage acetamitaphen cellulose. Aerosol composition (h)-(k) can be used in combination with standard aerosol disatorous agents, such as servicemanual, servicesecurity, Polysorbate 80, polyglycerol or oleic acid.

The results are given in the table.

1. Derivatives of indole of the formula (I)

in which R1represents hydrogen, halogen or methoxy;

R2represents hydrogen, halogen, methyl, ethyl or methoxy;

R3represents carboxy, tetrazolyl or CONHSO2R4where R4represents methyl, ethyl, phenyl, 2,5-dimethylisoxazole or trifluoromethyl;

T represents-CH2- or-SO2;

ring a is 3-chlorophenyl, 4-chlorophenyl, 3-triptoreline, 3,4-dichlorophenyl, 3,4-differenl, 3-fluoro-4-chlorophenyl, 3-chloro-4-forfinal or 2,3-dichloride-5-yl,

or its pharmaceutically acceptable salt or ester.

2. Connection on p. 1, in which ring a is 3-chlorophenyl, 4-chlorophenyl, 3-triptoreline, 3,4-dichlorophenyl, 3,4-differenl, 3-fluoro-4-chlorophenyl or 3-chloro-4-forfinal.

3. Connection on p. 2, in which ring a is 3,4-dichlorophenyl, 3-fluoro-4-chlorophenyl or 3-chloro-4-forfinal.

4. Connection on p. 1, in which ring a is 3,4-dichlorophenyl, 2,3-galactose-CH2-.

6. The compound according to any one of the preceding paragraphs, in which R3is carboxypropyl.

7. Connection on p. 1, in which in the compound of formula (I) R1is hydrogen, R2is hydrogen, R3is carboxypropyl, T represents-CH2-, and ring a is 3,4-dichlorophenyl or 3-chloro-4-forfinal, or its pharmaceutically acceptable salt or ester.

8. The method of obtaining the compounds of formula (I) under item 1, where R3means carboxy, which involves the reaction of compounds of formula (II)

in which Rand represents thecarboxy, or protected form;

Rbrepresents hydrogen or a suitable hydroxyamino group;

R1and R2represent groups defined in paragraph 1, with a compound of formula (III)

in which T and ring a have the meanings given in paragraph 1;

L represents a group to delete,

and then, if necessary, carry out the removal of any protective group, or transferred in pharmaceutically acceptable salt or ester.

9. Pharmaceutical composition comprising a compound according to any isrim chemoattractant protein-1 or RANTES (regulated on activation, expressed and secretory normal T-cells).

10. The compound according to any one of paragraphs.1-7 for use in preparation of medicines for the treatment of diseases mediated macrophage chemoattractant protein-1 or RANTES (regulated upon activation, expressed and secretory normal T-cells), such as an inflammatory disease.

 

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< / BR>
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< / BR>
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< / BR>
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