Condensed pyrrolo(2,3-c)carbazole-6-ones that enhance the activity of gamma-interferon

 

(57) Abstract:

The invention relates to novel condensed pyrrolo (2,3-C)carbazole-6-Onam represented by the General formulas (I) and (II).

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where R1- H, C1-C4alkyl;

R2- H, C1-C8alkyl;

R3and R4each independently - H, HE, C1-C8alkyl;

R5- H, C1-C8alkyl, C1-C8alkenyl, optionally substituted CF3;

X is-N-, -O-, -S-, alkylene with 1-3 carbons. The compounds I and II patentiert activity of human IFN in inducing the expression of MHC on the surface of the receptive cells are able to inhibit viral growth and/or tumor. 12 C.p. f-crystals, 7 ill., table 2.

The invention relates to pharmaceutical compounds, referred to in this patent description as "condensed pyrrolo[2,3-C]carbazole-6-ones". In addition, the disclosed methods of obtaining these compounds and methods of their use.

Human gamma-interferon (IFN-) is a natural human immunoregulatory protein. It is established that it is a tool that is effective in the treatment of tumors and viral infections in humans. The exact mechanism by which IFN - mu of two mechanisms: (1) acting directly on the cell, infected by the virus and the cell tumors and/or (2), first activating cells of the immune system, which then destroy infected with a virus a cell or a cell tumors [Interferons and other Regulatory Cytokines, E. De Maeyer and J. De Maeyer-'guignard, John Wiley & Sons, New York (1988)].

One of the manifestations stimulated the immune system is enhanced expression on the surface of immune cells protein major histocompatibility complex (MHC). The MTL consists of genes of class I, II and III, which encode the corresponding proteins of class I, II and III. Proteins of class I and II are located on the surface of cells and are involved in controlling the immune response, while class III proteins appear in the serum and are not involved in controlling the immune response. Proteins of class I and II (are) on antigen presenting cells such as monocytes. Lymphocytes, dendritic cells that present foreign antigens to T lymphocytes with subsequent destruction of cells containing a foreign antigen. Enhanced protein expression of class I and II required for the immune system to save the animal infected with the virus cells and increase the production of specific antibodies. IFN - is one of the major regulators of the immune response Blagodat IFN - is the increase in class I proteins on virus infected cells (virusinfizierovannah cells). Virusinfection cell is synthesized on its cell surface viral antigen receptor T cells to cytotoxic T cells (CD4 cells), followed by destruction of infected virus cells using cytotoxic T cells.

An example of the usefulness of MHC II increase with IFN - is the increase in protein class II on monocytes. Monocytes can digest invading (amazing) microorganisms and class II proteins on the surface of monocytes, presenting peptides derived from the invading microorganism. These peptides, proteins held class II, are present on the receptor of the T cells on the cells T-helper cells (CD8) with subsequent secretion of lymphokines CD8 cell. Secreted lymphokines cause proliferation of antibody synthesis In lymphocytes that synthesize large amounts of antibodies against the invading microorganism.

From the above examples you can see that the connection that enhances IFN - induction of MHC molecules, could be used in combination with IFN - a for the treatment of infections caused by microorganisms. This connection will allow you to reduce the dose of IFN-successfully providing the same therapeutic effect as in the case of the and messages about connections, which potentiate IFN--induced MHC expression [Coutinho, G. C., Dudrieu-Trautmann, O. , Strosberg, A. D. and Couraud, P. O. , Catecholamines Stimulate the IFN--induced Class II MHC Expression on Bovine Brain Capillary Endothelial Cells, J. Immunol. 147, 2525-2529 (1991); Zhu, J., Mix, E., Olsson, T. and Link, H. , "Influence of Ion Channel Modulation of in Vitro Interferon - Induced MHC Class I and II Expression on Macrophages", Immunopharmacology and Immunotoxicology, 17, 109-136 (1995); Mothes, T., Bendix, U., Pfannschmidt. C. and Lehmann. I., "Effect of Gliadin and Other Food Peptides on Expression of MHC Class II Molecules by HT-29 Cells, Gut, 36, 548-552 (1995)].

The invention

Here is disclosed a new class of compounds represented by formulas I and II, which we refer to as condensed pyrrolo[2,3-C]carbazole-6-ones.

THE FORMULA I

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FORMULA II

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Components of the substituents defined below. Preferred methods of obtaining these compounds are disclosed below.

We found that our connection condensed pyrrolo[2,3-C]carbazole-6-ones (numbering designated for K-a and-s proposed by Moody et. al, J. Org. Chem., 1992, 57, 2105) potentiate the activity of the human IFN in inducing the expression of MHC on the surface of the receptive (sensitive) cells. The compounds of this invention demonstrate the ability to enhance the effectiveness of the immune system, and this, in turn, successfully provides amplification inhibe the compounds of this invention are useful for augmentation, preferably, neurotrophin-3 (NT-3) activity.

Figure 1 is a histogram showing the increase in IFN--induced expression of HLA-DR MHC II molecules using pyrrolo[2,3-C]-carbazole-6-ones of this invention.

Figure 2 represents, in General terms, the scheme of chemical synthesis pyrrolo[2,3-C]carbazole-6-ones of formula I, section V (A)-(D).

Figure 3 is a scheme showing the synthesis of intermediates for pyrrolo[2,3-C]carbazole-6-ones.

Figure 4 presents a diagram of an alternative synthesis of intermediate for pyrrolo[2,3-C]carbazole-6-ones.

Figure 5 is a scheme showing the synthesis pyrrolo[2,3-C]carbazole-6-ones of formula II.

Figure 6 is a scheme showing the synthesis pyrrolo[2,3-C]carbazole-6-ones, in which X represents C=O from the corresponding compounds in which X is CH2.

Figure 7 is a scheme showing the synthesis pyrrolo[2,3-C]carbazole-6-she's from the intermediate product 11.

II. Condensed pyrrolo[2,3-C]carbazole-6-ones

The new compounds of this invention, which are referred to as condensed pyrrolo[2,3-C]carbazole-6-about derivatives represented by the following formula is La, arylalkyl, heteroaryl, heteroaromatic; C(= O)R9where R9represents alkyl with 1-4 carbons or aryl; (CH2)nOR9where n represents an integer of 1-4; OR10where R10represents H or alkyl with 1-4 carbons; (CH2)nOR SIG14where R14represents the amino acid residue after removal of hydroxyl carboxyl group; OR14, NR7R8; (CH2)nNR7R8and O(CH2)nNR7R8; and either

(1) R7and R8independently represent H or alkyl with 1-4 carbons; or

(2) R7and R8joined together with formation of a linking group of General formula -(CH2)2-X1(CH2)2- where X1represents O, S or CH2;

b) R2selected from the group consisting of H, SO2R9, CO2R9C(=O)R9, alkyl with 1-8 carbons, alkenyl with 1-8 carbons, quinil with 1-8 carbons and monosaccharide of 5-7 carbons, in which each hydroxyl group of the specified monosaccharide is independently selected from the group consisting of unsubstituted hydroxyl and fragment replaces the specified hydroxyl group, selected from H, alkyl with 1-4 carbon, alkylcarboxylic with 2-5 ug is kinil with 1-8 carbons is unsubstituted; or

2) each alkyl with 1-8 carbons, alkenyl with 1-8 carbons or quinil with 1-8 carbons independently substituted by 1-3 groups selected from the group consisting of aryl with 6-10 carbon, heteroaryl, F, Cl, Br, I, CN, NO2HE, OR9, O(CH2)nNR7R8, OCOR9, OCONHR9, O-tetrahydropyranyl, NH2, NR7R8, NR10COR9, NR10CO2R9, NR10CONR7R8, NHC(= NH)NH2, NR10S02R9; S(O)yR11where R11represents H, alkyl with 1-4 carbons, aryl with 6-10 carbon or heteroaryl, and y is 1 or 2; SR11, CO2R9, CONR7R8CHO, COR9CH2OR7CH2OR9CH= NNR11R12CH=NOR11CH=NR9CH=NNHCH(N=NH)NH2; SO2NRl2R13where or

(la) R12and R13independently represent H, alkyl with 1-4 carbons, aryl with 6-10 carbon or heteroaryl; or

(2A) R12and R13joined together with formation of a -(CH2)2-X1-(CH2)2linking group;

PO(OR11)2, OTHER14, NR10R14, OR14and monosaccharide of 5-7 carbons, in which each hydroxyl group of the specified monosaccharide is independently chosen, is selected from H, alkyl with 1-4 carbon, alkylcarboxylic with 2-5 carbons, and alkoxy with 1-4 carbons;

(C) each R3and R4independently selected from the group consisting of H, aryl with 6-10 carbon, heteroaryl, F, Cl, Br, I, CN, CF3, NO2HE, OR9O(CH2)nNR7R8, OCOR9, OCONHR9, NH2, (CH2)nOR SIG9, (CH2)nOR10, (CH2)nOR14, OR14, OTHER14, NR7R8, NR7(CH2)nNR7R8, NR10COR9, NR10CONR7R8, SR11, S(O)yR11,

CO2R9, COR9, CONR7R8CHO, CH= NOR11CH=NR9CH=NNR11R12, (CH2)nSR9, (CH2)nS(O)yR9; CH2SR15where R15represents alkyl with 1-4 carbons; CH2S(O)yR14, (CH2)nNR7R8, (CH2)nOTHER14, alkyl with 1-8 carbons, alkenyl with 1-8 carbons and quinil with 1-8 carbons; or

1) each alkyl with 1-8 carbons, alkenyl with 1-8 carbons or quinil with 1-8 carbons is unsubstituted or

2) each alkyl with 1-8 carbons, alkenyl with 1-8 carbons or quinil with 1-8 carbons is the independence of Anila with 1-8 carbons and quinil with 1-8 carbons, and either

1) each alkyl, Alchemilla or Alchemilla group is unsubstituted; or

2) each alkyl, Alchemilla or Alchemilla group is substituted by 1-3 groups selected from the group consisting of F, Cl, Br, I, CN, CF3, NO2HE, OR9, (CH2)nNR7R8, OCOR9, OCONHR9, NH2, (CH2)nOR9, (CH2)nOR14,

NR7R8, NR7(CH2)nNR7R8, NR10COR9, NR10CONR7R8, SR11, S(O)yR11, CO2R9, COR9, CONR7R8CHO, CH=NOR11CH=NR9CH=NNR11R12, (CH2)nSR9, (CH2)nS(O)yR9CH2SR15CH2S(O)yR14, (CH2)nNR7R8and (CH2)nOTHER14;

e) X is selected from the group consisting of-N-, -O-, -S-, -S(=O)-, -S(=O)2- alkylene with 1-3 carbons, -C(=O)-, -C(R2)=C(R2)-, -C(R2)2-, -CH=CH-, -CH(OH)-CH(OH)-, -C(=NOR11)-, -S(OR11)(R11)- -C(=O)CH(R15)-, -CH(R15)C(=O)-, -CH2-Z-, -Z-CH2-, -CH2ZCH2- where Z is selected from the group consisting of-C(OR11)(R11)-; O -, S -, C(=O), NR11; and alkylene with 1-3 carbons, Thames 14, R15, phenyl, naphthyl and arylalkyl with 7-14 carbons.

Used herein, the term "aryl" means a monocyclic or polycyclic aromatic group, including, for example, phenyl, raftiline, biphenylene and xylylene group. Aryl groups can be unsubstituted or substituted, for example, alkyl groups and Halogens. The Halogens include fluorine, chlorine, bromine and iodine. Preferred are aryl groups containing 6 to 10 carbons. Phenyl and raftiline groups are especially preferred.

Used herein, the term "heteroaryl" means aryl part, which contains at least one basic nitrogen atom and 0-4 heteroatoms selected from O, S and N. Examples of heteroaryl groups include pyrrolyl, pyranyl, tiopronin, furyl, imidazolyl, pyridyl, thiazolyl, triazinyl, phthalimido, indolyl, perineal and benzothiazolyl.

Used here with reference to the definition of R14the term "amino acid" means a molecule that contains both amino group and carboxyl group. It includes "amino acid", which has its usual meaning as a carboxylic acid, which is amidofunctional on the carbon adjacent to the carboxyl group. -The amino acid is water", defined here as two amino acids connected by peptide bond. Thus, components of the dipeptides are not limited to the amino acids and can be any molecule that contains both amino group and carboxyl group. Preferred are amino acids, dipeptides, such as lysyl-alanine, and aminoalkanoic (alkanoic) acids with 2 to 8 carbons, such as 3-dimethylaminomethylene acid.

Preferred "alkyl", "alkeneamine" and "alkyline" groups contain 1-4 carbon atoms.

Preferred R1groups include H, alkyl with 1-4 carbons, substituted or unsubstituted phenyl, OR10and(CH2)nNR7R8. Preferred phenyl substituents include alkyl with 1-4 carbons and halogen. The most preferred N.

Preferred R2groups include H, C(=O)R9, alkyl with 1-8 carbons and the alkyl with 1-8 carbons, substituted one OR9HE OCOR9, NR7R8, NH2, NR10COR9or NR10R14group. The most preferred N.

Preferred R3and R4groups include H, halogen, CN, IT, OR9, OR14, NH2, NR7R8, (CH2)n8. The most preferred N.

Preferred R5groups include h, alkyl with 1-4 carbon. The most preferred N.

Preferred X groups include-N-, -O-, -S-, alkylene with 1-3 carbons, -C= O-, -CH2-Z - and-Z-CH2-. Most preferred are N-, -O-, -S -, and-CH2group.

III. Use condensed pyrrolo[2,3-C]carbazole-6-ones

We offer condensed pyrrolo[2,3-C]carbazole-6-ones showed a number (a panoply important functional activities, which find application in a number of areas (settings), including both research and therapeutic area. For ease of presentation and in order not to limit the range of applications for which these compounds can be characterized preferred activity condensed pyrrolo[2,3-C] carbazole-6-ones can be generally described as follows:

A. Amplification of IFN - induction of MHC molecules;

C. Potentiation of the function and/or survival of cells that are sensitive to trophic factor.

The increased IFN - induction of MHC molecules may preferably be set using the cell line human monocytes, from which, the od inventory number (accession number ATCC TIB-202. IFN-known to induce the expression of three MHC II heterodimers. HLA-DP, HLA-DQ and HLA-DR; TNR-cells. Potentiation of the function and/or survival of cells that are sensitive to trophic factors such as nerve cells of origin (kind of related to the nerve cells), may preferably be set using the analysis of cultivated choline, acetyltransferase ("ChAT") of the spinal cord (a cultured spinal cord choline acetyltransferase ("ChAT") assay).

Used herein, the term "potentiation" when it is used for modification of the terms "function" and "survival" means a positive change or modification. Potentiation, which is positive, it is also called as "increase" or "increasing".

Used herein, the terms "increase" or "increasing", being used for modification of the terms "function" and "survival" means that the presence of condensed pyrrolo[2,3-C]carbazole-6-it has a relatively higher effect on the function and/or survival of cells that are sensitive to trophic factor, or, in the case of IFN-a on the induction of MHC molecules, than the comparative cell, not presents condensed PIM", "neural cell" includes, but is not limited to a heterogeneous population of neuronal types with singular or multiple transmitters and/or singular or multiple functions; preferably the neurons are cholinergic neurons. Used herein, the term "cholinergic neuron" refers to the neurons of the Central nervous system (CNS CNS) and peripheral nervous system (PNS PNS), whose neurotransmitter is acetylcholine; illustrative examples are the neurons of the basal forebrain and spinal cord.

Used herein, the term "trophic factor" is a molecule that directly or indirectly (indirect) effect on the survival or function of cells that are sensitive to trophic factor. Examples of trophic factors include ciliary neurotrophic factor Ciliary Neurotrophic Factor (CNTF), the main growth factor of the fibroblast (basic Fibroblast Growth Factor (bFGF), insulin and insulin-like growth factors (insulin and insulin-like growth factors (e.g., IGF-I, IGF-II, IGF-III), interferons, interleukins, cytokines, and neurotrophins, including growth factor nerve (Nerve Growth Factor (NGF), Neurotrophin-3 (NT-3), Neurotrophin-4/5 (NT-4/5) and neurotrophin factor obtained from the brain (Brain Derived Neurotrophic Factor (BDNF).

Used herein, the terms "activity trophic factor" and "activity induced by trophic factor" is defined as any response that directly or indirectly (indirect) is a consequence of the binding of trophic factor (for example, NT-3) with a cell comprising a receptor trophic factor.

Used here, the expression "activity trophic factor" and "activity induced by trophic factor", the term "trophic factor" includes both endogenous and exogenous trophic factors, where "endogenous" refers to trophic factor already present, and "exogenous" refers to a trophic factor that is added to the system. By definition, the activity induced by trophic factor" includes activity induced (1) endogenous trophic factors; (2) exogenous trophic factors, and (3) a combination of endogenous and exogenous trophic factors.

A. Amplification of IFN - induction of MHC molecules

Compounds of the invention of Maggi and some tumors. However, due to side effects, limiting its dose, the use of IFN - limited. In immunocompromise situation, for example in a patient with confirmed viral infection, increased IFN - mediated immune response would be beneficial, provided that IFN - induces the expression of MHC molecules. Therefore, the compounds of this invention, which increase the ability of endogenous IFN - or introduced exogenous IFN - induced MHC expression, are useful.

As described in detail in section V below, the ability of condensed pyrrolo[2,3-C] carbazole-6-it is to increase IFN - MHC molecules preferably evaluated using TNR-1 cell line. This cell line confirmed the expression of MHC II heterodimer HLA-DR. Comparison of the expression of HLA-DR in the presence of IFN - and IFN - plus one or more condensed pyrrolo[2,3-C] carbazole-6-ones of this invention provides a fast and effective way of determining gain IFN - induction of MHC molecules and can be evaluated.

The compounds of this invention can be used to develop in vitro models of increasing the expression of MHC molecules, functions, identification or other screening of synthetic compounds that have activity, and therefore research, aimed at research, purification and identification of molecular targets associated with functional responses. For example, by radiolabelling the condensed pyrrolo[2,3-C] carbazole-6-it is associated with a specific cellular function (e.g., HLA-DR induction), target (as such, objectively existing), which binds condensed pyrrolo[2,3-C]carbazole-6-he can be identified, isolated and purified to identify (characterization). In the following example, the condensed pyrrolo[2,3-C]carbazole-6-it can be used as skaniruyushchego tool for the detection means, which have a marginal activity, but when combined with at least one disclosed condensed pyrrolo[2,3-C]carbazole-6-one are able to enhance IFN - induction of MHC molecules. Because the proposed condensed pyrrolo[2,3-C]carbazole-6-ones used to enhance IFN - induction of MHC molecules, offer connections to success may be proposed for use as therapeutic agents. This increase has implications for immunocompromised patient.

C. Potentiation of the function and/or survival of cells that are sensitive to trophic facto is selenia function and/or survival of cells of neural origin. Condensed pyrrolo[2,3-C]carbazole-6-ones can be used individually or with other condensed pyrrolo[2,3-C]carbazole-6-areas, or in combination with other useful molecules, such as indolocarbazole, which also have the ability to potentiate the function and/or survival of the specified cells. In situations where the condensed pyrrolo[2,3-C] carbazole-6-it is believed, increases biological activity, such as neurotrophic activity, exogenous neurotrophins, such as NT-3, can be used in conjunction with the condensed pyrrolo[2,3-C] carbazole-6-one.

A number of neurological disorders are characterized by neural cells, which are lost, damaged with functional disorders, are axonal degeneration at risk of dying, and so on, These disorders include, but are not limited to: Alzheimer's disease; motor neuron disorders (e.g., amyotrophic lateral sclerosis); Parkinson's disease; cerebrovascular disorders (e.g. multiple sclerosis, peripheral neuropathy (e.g., neuropathy, affecting DRG neurons in peripheral neuropathy associated with chemotherapy); disorders caused vozbuditelno of penetrating into the brain or brain injuries or spinal cord.

As described in section V below, the ability of condensed pyrrolo[2,3-C] carbazole-6-it is to enhance the function and/or survival of cells of neural origin can be determined using the analysis of ChAT activity in the basal brain. ChAT catalyzes the synthesis of acetylcholine is a neurotransmitter and it is regarded as an enzymatic marker for functional cholinergic neuron. Functional neuron is also able to survive. The increase of neurotrophin, such as NT-3, can be determined by comparing the functional activity of neurotrophin in the presence or absence of condensed pyrrolo[2,3-C]carbazole-6-it.

Pharmaceutically acceptable salts of condensed pyrrolo[2,3-C]carbazole-6-ones are also covered by the scope of the invention. Used herein, the term "pharmaceutically acceptable salt" means a salt accession inorganic acid such as hydrochloride, sulfate, and phosphate, or a salt of the accession of organic acids, such as acetate, maleate, fumarate, tartrate and citrate. Examples of the pharmaceutically acceptable metal salts are alkali metal salts such as sodium salt and potassium, salts of alkaline earth metals such as magnesium salt and sole and salt Tetramethylammonium. Examples of pharmaceutically acceptable salts of the accession of organic amines are salts with morpholine and piperidine. Examples of pharmaceutically acceptable salts of joining amino acids are salts with lysine, glycine and phenylalanine.

Here, the compounds can be formulated in pharmaceutical compositions by mixing with pharmaceutically acceptable nontoxic excipients and carriers. Such compositions can be obtained for use in parenteral administration, particularly in the form of liquid solutions or suspensions; or oral introduction, in particular, in the form of tablets or capsules; or intranasally, particularly in the form of powders, nasal drops, or aerosols; or dermal application using, for example, transdermal bandages.

The composition is usually applied in unit dosage form and can be prepared by any method known in the pharmaceutical field, for example, described in Remington's Pharmaceutical Sciences (Mack Pub. Co., Easton, PA, 1980). Formulations for parenteral administration may contain as standard excipients sterile water or saline, polyalkylene glycols such as polietilenglikol degradiruem lacheny polymer, lactide/glycolide copolymer, or polyoxyethylene-polyoxypropylene copolymers may be useful excipients for controlled release of active compounds. Other potentially useful systems for parenteral delivery of these active compounds include particles of ethylene-vinyl acetate copolymer, osmotic pumps, implantable infusion system, and liposomes. Compositions for administration by inhalation contain as excipients, for example lactose, or they can be aqueous solutions containing, for example, polyoxyethylene-9-lauric ether, glycocholate and dezoksiholatom, or to provide oil solutions for administration in the form of nasal drops, or can be in the form of a gel to be applied intranasally. Formulations for parenteral administration may also include glycocholate to cheek (buccal) administration, salicylate for rectal injection or citric acid for vaginal administration. Formulations for transdermal bandages preferably represent a lipophilic emulsion.

The compounds of this invention can be used as an active agent in the pharmaceutical composition. Alternative they can AMI growth which facilitates the potentiation of NT-3, factors such as disclosed in US Patent No. 5468872 and International Publication No. WO 95/07911 (publication date: March 23, 1995).

The concentration of the compounds of this invention in therapeutic composition can vary. Concentration generally depends on factors such as the total dose of the medicinal product subject to introduction, the chemical characteristics (e.g., hydrophobicity) of the compounds used and route of administration. The compounds of this invention are usually applied in an aqueous physiological buffer solution containing about 0.1-10 wt.%/about. compounds for parenteral use. Range typical dose is from about 1 μg/kg to 1 g/kg body weight per day; a preferred dose range is from about 0.01 mg/kg to 100 mg/kg of body weight per day. The preferred dosage of drug to be used, apparently, depends on variables such as the type and extent of progression of the disease or disorder, the overall health status of the particular patient, the relative biological efficacy of the selected compound and the composition of the filler and the way of its introduction.

IV. A General description of the methods cinesouth two ways of synthesis. In method A (Fig.2) derivative of 2-(aryl) or 2-(heteroaryl)indole (1-4), which is either unsubstituted or substituted with 4-7 carbon (inclusive) indole rings (R3), or substituted or unsubstituted (hetero)aryl position (R4), is subjected to the interaction with maleimide in the presence of a catalyst, such as triperoxonane acid (TFA, TFA) to obtain the derivatives of condensed pyrrolopyrazole-6-it formula I (examples I-IV). To implement the reaction can also be used for more catalysts are Lewis acid, such as SnCl4, AS3, EtAlCl2or Et2AlCl. The reaction can also be carried out in a solvent such as TFA, toluene, CH2C12or 1,2-dichloroethane.

Biaryl indole intermediate 2,2'-beingal 1 (X=N, R2, R3, R4=H), 2-(2-furyl)indole 2 (X=0, R2, R3, R4=H) and 2-(benzothiazyl) indole 3 (X=S, R2, R3, R4= H) can be obtained using standard published methods (Hudkins, R. L., et al., J. Org. Chem., 1995, 60, 6218) (Fig.2). Derivatives of 2(2-Indenyl) indole 4 (Fig.3) (X=CH2, R2, R3, R4=H) or 2-(2-indenyl)indole, substituted R3or R4group, can be obtained by the interaction of 1-carboxy-2-(tribe, 995, 60, 6218), 2-bromoindene 6 (J. Org. Chem., 1982, 47, 705) or 2-bromoindene, substituted R4group. Alternative derivatives of 2-(2-indenyl) indole 4 (X=CH2, R2, R3, R4= H) or 2-(2-indenyl)indole, substituted R3or R4group, can be obtained by the interaction of the lH-indole or its derivative containing R3group protected in the form of lithium indole-1-carboxylate intermediate (Tetrahedron Lett. 26: 5935 (1985)), then treated with a strong base, such as t-BuLi, then alkylate the corresponding 2-indanone or 2-indanedione, substituted R4the group receiving the corresponding tertiary alcohol 7 (Fig.4). Received tertiary alcohol 7 is treated with dilute acid (for example, 2N model HC1 in acetone) to give the corresponding 2-(2-indenyl)indole 4 (US Patent 5475110). Using other benzocycloheptene-2-ones, such as 2-tetralone, in the sequence of reactions presented on figure 4, we can obtain 2-(2-(3,4-dihydronaphtho)indole 8 (X=CH2CH2, R2, R3, R4=H) (U.S. Pat. USA 5475110). You can use the method catalyzed by palladium cross-combinations (Stille reaction) to obtain other derivatives, for example, where X in the figure 3 has 1-3 carbon (inclusive), by combining 2-(tripto what tributylstannyl, getting 2-(2-benzocyclobutene)indoles.

The previously described source derived IH-indole in turn derived 1-substituted indole (R2not N) using standard methodology, for example by processing the IH-indole base and alkylating agent, receiving a 1-substituted indole. In these examples, a derivative of indole can be directly processed in a strong base (for example, t-BuLi, sec-BuLi, n-BuLi, lithium diisopropylamide) followed by alkylation derivative of 2-indanone, receiving the corresponding tertiary alcohol 7, which includes R2substituents in position one indole ring. A derivative of 2-(aryl) -, or 2-(heteroaryl)indole (1-3), 2-(2-indenyl)indole 4 or 2-(2-(1,2-dihydronaphtho)indole 8 can be converted into intermediates that contain R2substituents in position one indole ring by the method described above for the derivatives of indole.

Compounds of General formulas I and II, which contain R1group, no = N, can be obtained from the corresponding R1substituted maleimide (Fig.2, method A). Compounds of General formulas I and II, in which R1represents hydrogen, can be alkilirovanii in the presence of a base (for example, hydrides, alkoxides, hydroxides sheet tsepliaeva group, such as halogen, mesilate or toilet, derivatization condensed pyrrolopyrazole-6-it, which has R1group associated with the nitrogen of the lactam. Compounds of General formulas I and II, in which R5is hydrogen, can be converted into derivatives, where one or two R5groups can be introduced by treating the derivative condensed pyrrolopyrazole-6-it is one equivalent or an excess of a strong base (e.g., hydrides, alkoxides, hydroxides of alkaline or alkaline earth metals or organolithium compounds) with R5L, where L is tsepliaeva group, such as halogen, or by condensation with R5containing a ketone or a derivative of aldehydebase, getting derived condensed pyrrolopyrazole-6-it, which has one or two R5group. Derived from the reactions of ketone or aldehyde condensation usually give the vinyl derivative at R5.

Derivatives of indole get using standard methodology (US Patent 3976639; US Patent 3732245; The Chemistry of Heterocyclic Compounds. Indoles Parts One and Two; Houlihan Ed., Wiley-Interscience (1972)). Derivatives of 2-indanone can be obtained using previously described methods (see US Patent 4192888; US Patent 4128666; J. Am. Chem. Soc. 89:4525 (1990); Tetrahedron 45:1441 (1989); Synthesis 818 (1981)).

In the method (Fig.2) derivative of 2-(heteroaryl)indole (1-3) or a derivative of 2-(aryl)indole, such as 2-(2-indenyl)indole 4, is subjected to the interaction with ethyl CIS-cyanoacrylate in the presence of a catalyst, such as SiCl4, AS3, EtAlCl2Et2AlCl or TFA in CH2Cl2WITH2H4C12or toluene as solvent, receiving connection 6-exacerbate formula I of the invention.

The compounds of formula II are given as shown in Fig.5. Addition of 2-(aryl) -, or 2-(heteroaryl)indolizidine (Tetrahedron Lett., 1994, 35, 793; Tetrahedron Lett. , 1994, 35, 7123; Tetrahedron Lett., 1993, 35, 5955: Tetrahedron Lett., 1993, 34, 6245) to succinimido or derived R1substituted succinimide through reaction of the reformed (Reformatzsky) (Synthesis, 1975, 685), followed by dehydration usually gives compounds of General structure 9. Palladium-catalyzed ring-opening of (US Patent 5475110; Tetrahedron Lett. , 1993, 34, 8361) usually gives a connection pyrrolo[2,3-C]carbazole formula II.

Compounds in which X=(C=O) (General structure 10), obtained by oxidation of derivatives of General formula I (or II), where X=CH2using standard oxidizing reagents (e.g., SeO2, SGAs3, Na2SGAs7or IGOs2)2), C(R2)2WITH(OR11R11). Derivatives condensed pyrrolopyrazole-6-it, where X is S(=O) or S(=O)2can be obtained by oxidation of X=S derived in a manner analogous to X=(C=O).

EXAMPLES

V. Specific description of the methods of synthesis of

The following examples are provided for illustration and should not be construed as limiting in any way the scope of this invention.

A. Example I: 5H,7H,13H-Benzopyrano[2,3-a]pyrrolo[2,3-C]carbazole-6(6N)he (X=O; R1, R2, Rluxusni acid (2 ml) is heated in a sealed reaction vial at 125oC for 18 hours the Precipitated solid is collected, washed with TFA (TFA) and dried, receiving 150 mg (48%) of product. Recrystallization (THF, THF) gives a yellow-brown solid; T. pl. >300oC, MS (ES+) 312 (M+),1H NMR (D-d6) 4,00 (s, 2H), 4,13 (s, 2H), 7,12 (t, 1H), 7,37 (t, 2H), 7,45 (t, 2H), 7,71 (d, 1H), 7,95 (d, 1H), 8,54 (d, 1H), 10,12 (s, 1H), to 11.79 (s, 1H). Anal. the expect. for C20H12N2020,4 H2ABOUT; WITH, 75,18; N, ANDROID 4.04; N, 8,77. Found: C, 75,29, N, 3,86, N, 8,60.

C. Example II: 5H,7H,13H-Benzothieno[2,3-a]pyrrolo[2,3-C]carbazole-6(6N)he (X=S; R1, R2, R3, R4, R5=H)

A mixture of 2-(2-benzofuran)indole 2 (100 mg; 0.4 mmol) and maleimide (40 mg; 0.4 mmol) in triperoxonane acid (2 ml) is heated in a sealed reaction vial at 125oC for 16 hours the Precipitated solid is collected, washed with cold methanol and dried, receiving 80 mg (58%) of product. Recrystallization (THF) gives a yellow-brown solid; T. pl. >300oC, MS (ES+) 328 (M+)1H NMR (DMSO-d6) 4,10 (s, 2H), 7,20 (m, 1H), 7,40-of 7.60 (m, 4H), 8,10 (d, 1H), 8,80 (m, 1H), 10,86 (s, 1H), RS 11.80 (s, 1H). Anal. the expect. for C20H12N2SO0,5H2ABOUT; WITH, 71,20; N, 3,88; N, 8,30. Found: C, 70,86, N, 3,61, N, 8,39.

C. Example III: 5H,7H,N,13H-Indolo and 2-2'-byandala 1 (250 mg; 110 mmol) and maleimide (110 mg; 1.2 mmol) suspended in toluene (50 ml), add triperoxonane acid (0.5 ml). The solution is heated at the boiling temperature of phlegmy for 18 h, cooled to room temperature and concentrated to about 20 ml, the Solution is cooled in a bath with ice, the precipitated solid is collected, washed with cold ether and dried, receiving 150 mg (55%) of product. Purification using column chromatography (t:hexane; 2:1) gives a brown-yellowish-brown (brown-tan solid, so pl. >320oC, MS (ES+) 311 (M+),1H NMR (DMSO-d6) 4,00 (s, 2H), 7,17-7,22 (m, 2H), 7,37-7,42 (m, 2H), to 7.67 (d, 2H), 8,03 (d, 1H), 8,58 (d, 1H), 10,87 (s, 1H), 10,92 (s, 1H), 11,18 (s, 1H). Anal. the expect. for C20H12N2O20.5 H2O; C, 74,99; N, TO 4.41; N, 13,12. Found: C, 75,24, N, Was 4.02, N, Of 13.05.

(2) the Way IN

A mixture of 2-2'-byandala 1 (100 mg, 0.43 mmol) and ethyl CIS-cyanoacrylate (50 mg; 0.4 mmol) in methylene chloride (10 ml) add 25 ml of SnCl4. The mixture is stirred at room temperature for 30 minutes, the Suspension is cooled in a bath with ice, the solid is collected, washed with cold ether and dried, obtaining 36 mg (27%) of product. Purification using column chromatography (tO: hexane; 2:1) gives brown-Zheltov 2H), 7,17-7,22 (m, 2H), 7,37-7,42 (m, 2H), to 7.67 (d, 2H), 8,03 (d, 1H), 8,58 (d, 1H), 10,87 (s, 1H), 10,92 (s, 1H), 11,18 (s, 1H). This connection shows the spectral and analytical characteristics are identical to the characteristics obtained by method A.

D. Example IV: 5H,7H,N,13H-Indeno[2,3-a]pyrrolo[2,3-C]carbazole-6(6N)he (X=CH2; R1, R2, R3, R4, R5=N)

(1) the Method AND

Mix 2(2-indenyl) indole 4 (300 mg, 1.3 mmol) and maleimide (160 mg; 1.6 mmol) in triperoxonane acid (2 ml) is heated in a sealed reaction vial at 160oC for 18 hours the Solution is evaporated and the solid is dissolved in ethyl acetate, then washed with water and dried (gSO4) to give a brown solid product. The crude product chromatographic (silica gel, tOA:hexane; 1:1) to give the product, which is treated with THF and filtered. The concentrated residue is triturated with Meon, receiving the product: so pl. 275-280oC, MS (ES+) 310 (M+),1H NMR (DMCO-d6) 4,00 (s, 2H), 4,13 (s, 2H), 7 17 (t, 1H), 7,25-7,42 (m, 3H), 7,50 (d, 1H), 7,71 (d, 1H), 8,00 (d, 1H), of 8.37 (d, 1H), 10,75 (s, 1H), 11,33 (s, 1H), IR (KBR) 1650-1700 cm-1. Anal. the expect. for C21H14N2O0,7 H2O; C, 78,10; N, TO 4.81; N, 8,65. Found: C, 78,13, N, To 4.41, N, 8,10.

(2) the Way IN

Mix 2(2-indenyl) indole 4 (vannoy reaction vial at 120oC for 1 h, and then 4 h at 160oC. the Mixture is evaporated under reduced pressure and the residue triturated with ether. The obtained solid substance chromatographic (silica gel; tO:hexane; 1:1) to give 12 mg (12%) yellowish-brown solid product; so pl. 275-280oC, MS (ES+) 310 (M+). This connection shows spectral data similar to the data obtained by method A.

E. Example V: Strengthening condensed pyrrolo[2,3-C]carbazole-6-areas, called IFN - induction of MHC antigen HLA-DR

Cell line human, derived from human monocytes, TPR-1 (ATSS TIB 202), which responds to IFN-, which is used to demonstrate the increase of HLA-DR condensed pyrrolo[2,3-C] carbazole-6-areas.

TNR-1 cells are grown in RPMI 1640 medium containing 20 μm mercaptoethanol and 10% fetal calf serum at 37oC in an atmosphere with 5% CO2:95% air at 100% humidity. To determine the increase of HLA-DR by the compounds of the present invention cells either left untreated as a control treated only IFN - a at a concentration of 100 units/ml, or treated by the compounds of the invention at a final concentration of 1 μm for 30 min before the addition of IFN - if con is installed TNR-1 cells incubated at 37oC for 48 hours and then prepared for analysis of HLA-DR by flow cytometry. The induction of HLA-DR is carried out by standard procedures, as described in Interferons and Other Regulatory Cytokines, Edward De Maeyer and Jacqueline De Maeyer'guignard, Chapter 9, John Wiley & Sons, New York, 1988. Cells prepare for flow cytometry and analyzed for HLA-DR by flow cytometry, as described in Current Protocols in Immunology. Vol. 1, pages 5.0.1-5.8.8, John Wiley & Sons, 1994. One million cells from each treatment are collected by centrifugation and washed twice with phosphate-saline buffer solution (PBS). Cells re-suspended in 100 μl PBS containing 10 μg of purified rabbit IgG to block non-specific sites on the cell surface. After 20 min of incubation on ice, add 20 ál of anti-HLA-DR monoclonal antibodies labeled with fluorescent-labeled FITC, cells and left on ice for an additional 30 min to allow the antibody to contact HLA-DR. The cells are then washed 2 times with PBS and fixed in 0.5 ml of 0.5% paraformaldehyde. Fixed cells stored at 4oWith before analysis by flow cytometry.

Increased HLA-DR representative condensed pyrrolo[2,3-C] carbazole-6-areas shown in Fig.1. Increased HLA is be representative compounds at 1 μm (Fig.1). All of the representative compounds enhance the induction of HLA-DR with IFN - induction over only one IFN-, i.e. above 100%. Percentage increase over only one IFN - through the four compounds shown in table 1. For example, 2 μm, the connection section V(D) (example IV) enhances IFN - induction of HLA-DR by 60% compared IFN - one.

F. Example VI. NT-3 Potentiation of ChAT activity in the basal crop brain pyrrolo[2,3-C]carbazole-6-areas

The ability pyrrolo[2,3-C]carbazole-6-ones to potentiate the NT-3 activity in basal cultures brain determine, using choline acetyltransferase (ChAT) activity as a measure of cholinergic neuronal function or survival. These compounds, by themselves, have no action on ChAT activity. However, in the presence of NT-3 representative compounds give dozozavisimoe potentiation ChAT activity to levels greater than the levels that are achieved by using only NT-3. The results presented in table 2 represent the result of a single application of NT-3 and the connection to be tested, on the day of initiation of culture, showing a prolonged effect on the survival or function of cholinergic neurons of the basal golovnogo) compared to control untreated cultures. Test compounds themselves are basal (100%) level of activity (p<0.05 is statistically significant when compared with NT-3 activity by itself according to Dunnett t-statistic).

Specialists in the art it is obvious that various changes and modifications of the preferred variants of the invention and that such changes and modifications can be made without leaving the scope of the essence of this invention. So have in mind that the following claims cover all equivalent changes or modifications not beyond the nature and scope of the invention. Documents cited throughout the disclosure of this patent, incorporated herein as prior art.

The applicant received two more connections, the members of the circle of connections item 1:

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These compounds were synthesized as described in the application of techniques, methods and tools, and experiences to strengthen induced IFN - induction of MHC antigen HLA-DR showed activity at the level of the compounds of example I and II, respectively, and in the experience of NT-3 potentiation ChAT activity in basal cultures at the connection level I. with regard to properties, causing the same purpose tablenewline compounds. This further confirms the activity of two of the newly obtained compounds, which, as predicted by the applicant, was the corresponding required parameters.

1. Condensed pyrrolo[2,3-c] carbazole-6-it is represented by a formula selected from the group consisting of

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and

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where R1selected from the group consisting of H and alkyl with 1-4 carbons;

R2selected from the group consisting of H and alkyl with 1-8 carbons;

R3and R4each independently selected from the group consisting of H, HE and alkyl with 1-8 carbons;

R5selected from the group consisting of H, alkyl with 1-8 carbons and alkenyl with 1-8 carbons, optionally substituted CF3;

X is selected from the group consisting of-N-, -O-, -S - and alkylene with 1-3 carbon.

2. Connection on p. 1, where R1selected from the group consisting of H and alkyl with 1-4 carbon.

3. Connection on p. 1, where R2- N.

4. Connection on p. 1, where R3and R4each independently selected from the group consisting of H and HE.

5. Connection on p. 1, where R5selected from the group consisting of H and alkyl with 1-4 carbon.

6. Connection on p. 1, where X is selected from the group consisting of-N-, -O-, -S - is expressed by formula I.

9. Connection on p. 8, where R1selected from the group consisting of H and alkyl with 1-4 carbon.

10. Connection on p. 8, where R2selected from the group consisting of H and alkyl with 1-4 carbon.

11. Connection on p. 8, where R3and R4each independently selected from the group consisting of H and HE.

12. Connection on p. 8, where R5selected from the group consisting of H and alkyl with 1-4 carbon.

13. Connection on p. 8, where X is selected from the group consisting of-N-, -O-, -S -, and-CH2-.

 

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