KR20040025903A - Benzimidazole compounds for modulating ige and inhibiting cellular proliferation - Google Patents

Abstract

The present invention relates to small molecule inhibitors of IgE responses to allergens useful for the treatment of allergies and / or asthma or any disease in which IgE is pathogenic. The present invention also relates to a benzimidazole molecule that is a cell proliferation inhibitor and is useful as an anticancer agent.

Description

Benzimidazole compounds for regulating IgE and inhibiting cell proliferation {BENZIMIDAZOLE COMPOUNDS FOR MODULATING IGE AND INHIBITING CELLULAR PROLIFERATION}

Technical Field

The present invention relates to small molecule inhibitors of the IgE response to allergens useful for the treatment of allergies and / or asthma, or any disease in which IgE is pathogenic. The present invention also relates to small molecules that are proliferation inhibitors, which are useful as anticancer agents.

Description of the prior art

Allergies and asthma

An estimated 10 million people in the United States, about 5% of the total population, have asthma. The cost of asthma in the United States is estimated to exceed 6 billion. About 25% of asthma patients who need emergency care require hospitalization, and the largest one-way medical expenditure for asthma is inpatient hospital services (emergency care) costing more than $ 1.6 billion. The cost of prescription drugs increased 54% between 1985 and 1990 to $ 1.1 billion (see Kelly, Pharmacotheraphy 12: 13S-21S, 1997).

According to the National Ambulatory Medical Care Survey, 1% of all ambulatory care visitors are asthma, and the disease continues to be a major cause of children's absences. Despite improved understanding of this disease process and better drugs, asthma mortality and mortality rates continue to increase in the United States and around the world (US Department of Health and Human Services; 1991, publication no.91-3042). . As such, asthma is a major public health problem.

The pathophysiological process of developing asthma is inherently divided into two stages characterized by panting, chest tightness, and bronchial contraction causing breathing difficulties. First, the initial asthma response is triggered by allergens, stimulants, or exercise. Allergen-crosslinked immunoglobulin E (IgE) molecules bound to receptors on mast cells allow mast cells to release many pre-formed inflammatory mediators including histamine. An additional trigger is osmotic changes in airway tissue after exercise or inhalation of cold dry air. Secondly, the subsequent late response is characterized by the infiltration of activated eosinophils and other inflammatory cells into airway tissues and the presence of highly viscous mucus in the airways. Damage caused by this inflammatory response can cause the airways to “sensitize” or become sensitive, resulting in subsequent asthma symptoms even with smaller triggers.

Many drugs can be used to treat asthma relief. However, their efficacy varies greatly. The long-term major drugs in the treatment of asthma, the fast-acting β ₂ -adrenergic agonists, terbutaline and albuterol, act primarily primarily as bronchodilators. Newer long-acting β ₂ agonists, salmeterol and formoterol can reduce the bronchoconstriction component of the late response. However, since β ₂ -agonists do not have much anti-inflammatory activity, they are ineffective in bronchial hypersensitivity.

Many other drugs target specific aspects of early or late asthma reactions. For example, antihistamines, such as loratadine, inhibit early histamine mediated inflammatory responses. Some new antihistamines, such as azelastine and ketotifen, may have both anti-inflammatory and mild bronchodilator effects, but they currently lack established efficacy for treating asthma. Phosphodiesterase inhibitors such as theophylline / xanthine may attenuate late inflammatory responses, but there is no evidence that these compounds reduce bronchial hypersensitivity. Anticholinergic agents, such as ipratopinium bromide, used in the case of acute asthma to inhibit severe bronchial contraction, do not have an effect on early or late inflammation, do not have an effect on bronchial hypersensitivity, and therefore may essentially contribute to chronic treatment Can't.

Corticosteroid drugs such as budesonide are the most potent anti-inflammatory drugs. Anti-inflammatory mediators act by releasing inhibitors such as chromoline and nedocromyl and stabilizing mast cells, thereby inhibiting late inflammatory responses to allergens. Thus, chromosteroids as well as chromoline and nedocromil both reduce bronchial hyperresponsiveness by minimizing the sensitizing effects of inflammatory damage to the airways. Unfortunately, these anti-inflammatory agents do not cause bronchiectasis.

Several new agents have been developed that suppress certain aspects of asthma inflammation. For example, leukotriene receptor antagonists (ICI-204, 219, acolate) specifically inhibit leukotriene mediated action. Rukotriene is involved in the induction of both airway inflammation and bronchial contraction.

Thus, while many drugs are currently available for the treatment of asthma, these compounds mainly have temporary relief or have serious side effects. As a result, new treatment methods aimed at the underlying cause, rather than a series of staged symptoms, would be highly desired. Asthma and allergies are dependent on common IgE mediated events. Indeed, excess IgE production is generally known to be a fundamental cause of allergies, especially allergic asthma (Duplantier and Cheng, Ann. Rep. Med. Chem. 29: 73-81, 1994). As such, compounds that lower IgE levels may be effective in treating the underlying cause of asthma and allergies.

Current treatments do not eliminate excess blood IgE. The assumption that lowering plasma IgE can reduce allergic reactions has been confirmed by recent clinical results using chimeric anti-IgE antibodies, CGP-51901, and recombinant humanized monoclonal antibodies, rhuMAB-E25. Indeed, three companies, Tanox Biosystems, Inc., Genentech Inc., and Novartis AG allergens by neutralizing excess IgE Humanized anti-IgE antibodies to treat asthma (BioWorld® Today, February 26, 1997, p. 2) are being developed in collaboration. Tanox has already successfully tested the anti-IgE antibody, CGP-51901, which reduces the severity and duration of nasal symptoms of allergic rhinitis in a Phase II trial of 155 patients. Genentech recently published positive results from Phase II / III testing of 536 patients with the recombinant humanized monoclonal antibody, rhuMAB-E25 (BioWorld® Today, November 10, 1998, page 1). The antibody, ruhMAB-E25, administered by injection (up to 300 mg every two to four weeks, if necessary), requires that the patient require additional "rescue" drugs (antihistamines and anti-inflammatory agents) as compared to placebo. The number of days was reduced by 50%. More recently, Henry Milgrom et al. Of the National Jewish Medical and Research Center in Denver, Colorado has reported clinical results of rhuMAB-25 in moderate to severe asthma patients (317 patients in 12 weeks). , Intravenous injections every two weeks), and concluded that this drug would be a "solution" (New England Journal of Medicine, December 23, 1999). A Biologics License Application (BLA) for these products was submitted to the FDA in June 2000 by Novartis Pharmachemicals Corporation, Tanox Incorporated, and Genentech Incorporated. Positive results of the anti-IgE antibody test suggest that therapeutic strategies aimed at controlling IgE drop can be effective.

Cancer and Hyperproliferative Diseases

Cell proliferation is a normal process that is important for the normal functioning of most biological processes. Cell proliferation occurs in all living organisms and includes two major processes, nuclear fission (mitosis) and cytokinesis. Because organisms continue to proliferate and exchange cells, cell proliferation is essential to the health of healthy cells. Disruption of normal cell proliferation can lead to a variety of diseases. For example, hyperplasia of cells can lead to psoriasis, thrombosis, atherosclerosis, coronary heart disease, myocardial infarction, seizures, myofascial neoplasia, fibroid embolization or fibroids, and obstructive diseases of vascular grafts and transplant organs. Abnormal cell proliferation is most commonly associated with tumor formation and cancer.

Cancer is one of the highest causes of death in the United States and internationally. In fact, cancer is the second leading cause of death in the United States. According to the National Institute of Health, the annual cost of cancer is about $ 107 billion, which is $ 37 billion in direct health care costs, $ 11 billion indirect costs of lost productivity due to illness, and productivity losses due to premature death. Indirect costs include $ 59 billion. It is not surprising that considerable efforts are underway to develop new therapies and preventive measures to treat these destructive diseases.

Currently, cancer is being treated using a combination of surgery, radiation therapy and chemotherapy. Chemotherapy involves the use of chemical agents to disrupt the replication and metabolism of cancer cells. Chemotherapeutic agents currently used to treat cancer can be classified into five major groups, natural products and their derivatives, anthracyclines, alkylating agents, antiproliferative agents, and hormonal agents.

One embodiment of the invention discloses benzimidazole compounds that modulate IgE and inhibit cell proliferation. Benzimidazole compounds are conventionally known in the art, for example in European Patent 719,765 and US Pat. No. 5,821,258. Both documents disclose compounds containing active ingredients that act on DNA, but are structurally different from the benzimidazole derivatives of the present invention. Conventional compounds alkylate DNA and the literature does not disclose that the disclosed benzimidazole compounds modulate IgE or inhibit cell proliferation. In addition, the compounds described in both documents are described as anticancer agents, antiviral agents, or antibacterial agents. The antiallergic or anti-asthmatic properties of the benzimidazole compounds of the present invention have not been previously recognized. In addition, in describing the anticancer properties of benzimidazole compounds, these documents disclose chemotherapeutic agents which are DNA alkylating agents. Inhibition of cell proliferation using the compounds of the present invention has not been disclosed.

Summary of the Invention

The present invention discloses several compounds that are active in modulating the IgE response to allergens and other stimulatory stimulants. One compound disclosed for the treatment of diseases associated with excess IgE levels and / or abnormal cell proliferation has the formula:

In the above,

R is selected from the group consisting of H, C ₁ -C ₅ alkyl, benzyl, p-fluorobenzyl, and dialkylamino alkyl, and C ₁ -C ₅ alkyl is from the group consisting of linear, branched, or cyclic alkyl Selected;

R ₁ and R ₂ are independently H, alkyl, substituted alkyl, C ₃ -C ₉ cycloalkyl, substituted C ₃ -C ₉ cycloalkyl, polycyclic aliphatic groups, phenyl, substituted phenyl, naphthyl, substituted Selected from the group consisting of naphthyl, heteroaryl, and substituted heteroaryl, wherein the heteroaryl and substituted heteroaryl contain 1-3 heteroatoms, wherein the heteroatoms are independently composed of nitrogen, oxygen, and sulfur Selected from the group;

Substituted phenyl, substituted naphthyl, and substituted heteroaryl contain 1-3 substituents, wherein the substituents are H, halogen, polyhalogen, alkoxy group, substituted alkoxy, alkyl, substituted alkyl, dialkylamino Alkyl, hydroxyalkyl, OH, OCH ₃ , COOH, COOR ', COR', CN, CF ₃ , OCF ₃ , NO ₂ , NR'R ', NHCOR', and CONR'R ';

R ₃ and R ₄ are independently selected from the group consisting of H, alkyl, aryl, heteroaryl, and COR ′;

R 'is H, alkyl, substituted alkyl, C ₃ -C ₉ cycloalkyl, substituted C ₃ -C ₉ cycloalkyl, polycyclic aliphatic groups, phenyl, substituted phenyl, naphthyl, substituted naphthyl, heteroaryl , And substituted heteroaryl, wherein the heteroaryl and substituted heteroaryl contain 1-3 heteroatoms, and the heteroatoms are independently selected from the group consisting of nitrogen, oxygen, and sulfur;

X and Y are independently H, halogen, alkoxy, substituted alkoxy, alkyl, substituted alkyl, dialkylaminoalkyl, hydroxyalkyl, OH, OCH ₃ , COOH, CN, CF ₃ , OCF ₃ , NO ₂ , COOR '', CHO, and COR '';

R '' is C ₁ -C ₈ alkyl, wherein C ₁ -C ₈ alkyl is selected from the group consisting of linear, branched, or cyclic alkyl.

The aforementioned class of compounds may contain polycyclic aliphatic groups selected from the group consisting of adamantyl, bicycloheptyl, camphoryl, bicyclo [2,2,2] octanyl, and norbornyl.

The aforementioned class of compounds include pyridine, thiazole, isothiazole, oxazole, pyrimidine, pyrazine, furan, thiophene, isoxazole, pyrrole, pyridazine, 1,2,3-triazine, 1,2,4 -Triazine, 1,3,5-triazine, pyrazole, imidazole, indole, quinoline, isoquinoline, benzothiopine, benzofuran, parathiazine, pyran, chromen, pyrrolidine, pyrazolidine, It may contain heteroaryl and substituted heteroaryl selected from the group consisting of imidazolidine, morpholine, thiomorpholine, and the corresponding heterocyclic.

Specific compounds of class I according to the invention are also disclosed. These compounds are referred to as compounds I.1 to I.192, and their representative structures are as described below. In accordance with the present invention, other compounds have been disclosed that are used for the treatment of conditions associated with excess IgE levels and / or abnormal cell proliferation. The compound has the formula:

In the above,

R is selected from the group consisting of H, C ₁ -C ₅ alkyl, benzyl, p-fluorobenzyl, and dialkylamino alkyl, and C ₁ -C ₅ alkyl is from the group consisting of linear, branched, or cyclic alkyl Selected;

R ₁ and R ₂ are independently H, alkyl, substituted alkyl, C ₃ -C ₉ cycloalkyl, substituted C ₃ -C ₉ cycloalkyl, polycyclic aliphatic groups, phenyl, substituted phenyl, naphthyl, substituted Selected from the group consisting of naphthyl, heteroaryl, and substituted heteroaryl, wherein the heteroaryl and substituted heteroaryl contain 1-3 heteroatoms, wherein the heteroatoms are independently composed of nitrogen, oxygen, and sulfur Selected from the group;

Substituted phenyl, substituted naphthyl, and substituted heteroaryl contain 1-3 substituents, wherein the substituents are H, halogen, polyhalogen, alkoxy group, substituted alkoxy, alkyl, substituted alkyl, dialkylamino Alkyl, hydroxyalkyl, OH, OCH ₃ , COOH, COOR ', COR', CN, CF ₃ , OCF ₃ , NO ₂ , NR'R ', NHCOR', and CONR'R ';

R ₃ and R ₄ are independently selected from the group consisting of H, alkyl, aryl, heteroaryl, and COR ′;

R 'is H, alkyl, substituted alkyl, C ₃ -C ₉ cycloalkyl, substituted C ₃ -C ₉ cycloalkyl, polycyclic aliphatic groups, phenyl, substituted phenyl, naphthyl, substituted naphthyl, heteroaryl , And substituted heteroaryl, wherein the heteroaryl and substituted heteroaryl contain 1-3 heteroatoms, and the heteroatoms are independently selected from the group consisting of nitrogen, oxygen, and sulfur;

X and Y are independently H, halogen, alkoxy, substituted alkoxy, alkyl, substituted alkyl, dialkylaminoalkyl, hydroxyalkyl, OH, OCH ₃ , COOH, CN, CF ₃ , OCF ₃ , NO ₂ , COOR '', CHO, and COR '';

R '' is C ₁ -C ₈ alkyl, wherein C ₁ -C ₈ alkyl is selected from the group consisting of linear, branched, or cyclic alkyl.

The aforementioned class of compounds may contain polycyclic aliphatic groups selected from the group consisting of adamantyl, bicycloheptyl, camphoryl, bicyclo [2,2,2] octanyl, and norbornyl.

The aforementioned class of compounds include pyridine, thiazole, isothiazole, oxazole, pyrimidine, pyrazine, furan, thiophene, isoxazole, pyrrole, pyridazine, 1,2,3-triazine, 1,2,4 -Triazine, 1,3,5-triazine, pyrazole, imidazole, indole, quinoline, isoquinoline, benzothiopine, benzofuran, parathiazine, pyran, chromen, pyrrolidine, pyrazolidine, And heteroaryl selected from the group consisting of imidazolidine, morpholine, thiomorpholine, and the corresponding heterocyclic and substituted heteroaryl.

Specific compounds of class II according to the invention are also disclosed. These compounds were referred to as compounds II.1 to II.90 and their representative structures are shown below.

In accordance with the present invention, other compounds are disclosed that are used for the treatment of diseases associated with excess IgE levels and / or abnormal cell proliferation. The compound has the formula:

In the above,

R is selected from the group consisting of H, C ₁ -C ₅ alkyl, benzyl, p-fluorobenzyl, and dialkylamino alkyl, and C ₁ -C ₅ alkyl is from the group consisting of linear, branched, or cyclic alkyl Selected;

R ₁ and R ₂ are independently H, alkyl, substituted alkyl, C ₃ -C ₉ cycloalkyl, substituted C ₃ -C ₉ cycloalkyl, polycyclic aliphatic groups, phenyl, substituted phenyl, naphthyl, substituted Selected from the group consisting of naphthyl, heteroaryl, and substituted heteroaryl, wherein the heteroaryl and substituted heteroaryl contain 1-3 heteroatoms, wherein the heteroatoms are independently composed of nitrogen, oxygen, and sulfur Selected from the group;

Substituted phenyl, substituted naphthyl, and substituted heteroaryl contain 1-3 substituents, wherein the substituents are H, halogen, polyhalogen, alkoxy group, substituted alkoxy, alkyl, substituted alkyl, dialkylamino Alkyl, hydroxyalkyl, OH, OCH ₃ , COOH, COOR ', COR', CN, CF ₃ , OCF ₃ , NO ₂ , NR'R ', NHCOR', and CONR'R ';

R ₃ and R ₄ are independently selected from the group consisting of H, alkyl, aryl, heteroaryl, and COR ′;

R 'is H, alkyl, substituted alkyl, C ₃ -C ₉ cycloalkyl, substituted C ₃ -C ₉ cycloalkyl, polycyclic aliphatic groups, phenyl, substituted phenyl, naphthyl, substituted naphthyl, heteroaryl , And substituted heteroaryl, wherein the heteroaryl and substituted heteroaryl contain 1-3 heteroatoms, and the heteroatoms are independently selected from the group consisting of nitrogen, oxygen, and sulfur;

X and Y are independently H, halogen, alkoxy, substituted alkoxy, alkyl, substituted alkyl, dialkylaminoalkyl, hydroxyalkyl, OH, OCH ₃ , COOH, CN, CF ₃ , OCF ₃ , NO ₂ , COOR '', CHO, and COR '';

R '' is C ₁ -C ₈ alkyl, wherein C ₁ -C ₈ alkyl is selected from the group consisting of linear, branched, or cyclic alkyl.

The aforementioned class of compounds may contain polycyclic aliphatic groups selected from the group consisting of adamantyl, bicycloheptyl, camphoryl, bicyclo [2,2,2] octanyl and norbornyl.

The aforementioned class of compounds include pyridine, thiazole, isothiazole, oxazole, pyrimidine, pyrazine, furan, thiophene, isoxazole, pyrrole, pyridazine, 1,2,3-triazine, 1,2,4 -Triazine, 1,3,5-triazine, pyrazole, imidazole, indole, quinoline, isoquinoline, benzothiopine, benzofuran, parathiazine, pyran, chromen, pyrrolidine, pyrazolidine, It may contain heteroaryl and substituted heteroaryl selected from the group consisting of imidazolidine, morpholine, thiomorpholine, and the corresponding heterocyclic.

Specific compounds of class III according to the invention are also disclosed. These compounds are referred to as compounds III.1 to III.154, and their representative structures are described below.

For each chemical structure disclosed herein, the hydrogen atom on the heteroatom is omitted for clarity. Where open valences on heteroatoms are indicated, these valences are assumed to be filled with hydrogen atoms.

Also disclosed are methods for treating a disease state associated with excess IgE and / or abnormal cell proliferation (eg cancer) in a mammal. In one aspect, the method comprises administering to a mammal an pharmaceutical formulation of an IgE inhibitory or cytostatic amount comprising at least one benzimidazole compound of the small molecule group disclosed above.

Depending on the method of treatment, the small molecule IgE inhibitory compound may be administered with one or more additional agents that are active in reducing the symptoms associated with allergic reactions. In one embodiment, the small molecule inhibitor can be mixed with one or more additional active ingredients to form a pharmaceutical composition. Alternatively, small molecule inhibitors may be administered simultaneously with one or more additional active agents or together in accordance with different therapeutic dosage regimens.

One or more additional active ingredients include an immediate β ₂ -adrenergic agent selected from the group consisting of terbutalin and albuterol; Persistent β ₂ -adrenergic agents selected from the group consisting of salmeterol and formoterol; Antihistamines selected from the group consisting of loratadine, azelastine, and ketotifen; Phosphodiesterase inhibitors or leukotriene receptor antagonists.

In other embodiments, the benzimidazole compound may be administered with one or more additional active agents. Such active agents include antifungal agents, antiviral agents, antibiotics, anti-inflammatory agents, and anticancer agents. The anticancer agent is an alkylating agent (Romustine, Carmustine, Streptozosin, Mechloretamine, Mephalan, Urasyl Nitrogen Mustard, Chlorambucil, Cyclophosphamide, Iphosphamide, Cisplatin, Carboplatin Mitomycin Thiotepa Dakar Vagin procarbazine, hexamethyl melamine, triethylene melamine, busulfan, fibrobroman, and mitotan); Metabolic agents (methotrexate, trimetrexate pentostatin, cytarabine, ara-CMP, fludarabine phosphate, thioguanine, and 6-mercaptopurine); DNA cutters (bleomycin); Topoisomerase I activity inhibitors (topotecan irinotecan and camptothecin); Topoisomerase II activity inhibitors (daunorubicin, doxorubicin, idarubicin, mitosantron, teniposide, and etoposide); DNA binding agents (dactinomycin and mitramycin); And spindle inhibitors (vinblastine, vincristine, nabelvin, paclitaxel, and docetaxel).

In other embodiments, the benzimidazole compounds of the invention may be administered in combination with one or more other therapies. Such therapies include, but are not limited to, radiation therapy, immunotherapy, gene therapy, and surgery. For example, radiation therapy may be administered in conjunction with administration of the benzimidazole compound, or may be administered at any time before or after administration of the benzimidazole compound.

Small molecule IgE inhibitory compounds at a dose of about 0.01 mg to about 100 mg / kg body weight / day may be administered in divided doses daily.

Treating a disease state associated with excess IgE or abnormal cell proliferation in a mammal, comprising administering to the mammal a therapeutic formulation comprising a pharmaceutical formulation comprising one or more compounds selected from class I, class II, and / or class III A method is also disclosed.

The methods provided herein for treating diseases and processes mediated by unnecessary, uncontrolled, or abnormal cell proliferation include administering to a mammal a composition of the benzimidazole compound disclosed herein to inhibit cell proliferation. The method is particularly useful for preventing or treating tumor formation and progression. In one embodiment of the present invention, the disclosed compounds and methods are particularly useful for treating estrogen receptor positive and estrogen receptor negative type breast cancer.

Other variations within the scope of the invention may be better understood with reference to the following detailed description.

Brief description of the drawings

1 shows the inhibition of splenic cell proliferative response by compound I.82. Spleen cell cultures were established from naïve BALB / c mice and incubated for 4 days in the presence of stimuli and drugs. Cultures were pulsed with 3H-thymidine and collected.

Detailed Description of the Preferred Embodiments

The present invention relates to small molecule inhibitors of IgE useful for the treatment of allergies and / or asthma or any disease in which IgE is pathogenic. Inhibitors can affect the synthesis, activity, release, metabolism, degradation, clearance, and / or pharmacokinetics of IgE. Certain compounds disclosed herein can be identified by their ability to inhibit IgE in both ex vivo and in vivo assays. The compounds disclosed herein are also useful for the treatment of diseases associated with abnormal cell proliferation, including but not limited to tumor formation, and other proliferative diseases such as cancer, inflammatory diseases, and circulatory diseases. Development and optimization of clinical therapies can be monitored by those skilled in the art with reference to the in vitro and in vivo assays described below.

In vitro assay

The system starts with antigen sensitization in vivo and measures the secondary antibody response in vitro. The basic protocol is documented and optimized for a range of parameters, including: antigen dose for sensitization and sensitization over time, in vitro cultured cells, inducing a second in vitro IgE (and other immunoglobulin) response Fetal calf serum (FBS) placement to enable specificity of antigen concentration for, in vitro optimal IgE response, importance of sensitized CD4 + T cells and hapten-specific B cells, and ELISA assay for IgE (see Marcelletti) and Katz, Cellular Immunology 135: 471-489, 1991; incorporated herein by reference).

The actual protocol used for this project is adapted for more efficient analysis. BALB / cByj mice were immunized by intravenous injection with 10 μg DNP-KLH adsorbed on 4 mg alum and killed 15 days later. The spleen is homogenized in a tissue mill and washed twice and maintained in DMEM supplemented with 10% FBS, 100 U / ml penicillin, 100 μg / ml streptomycin and 0.0005% 2-mercaptoethanol. Spleen cell cultures are established with or without DNP-KLH (10 ng / ml) (2 to 3 million cells / ml in 4 96 well plates, 0.2 ml / well). Test compounds (2 μg / ml and 50 ng / ml) are added to spleen cell cultures containing antigen and incubated at 37 ° C. for 8 days under 10% CO ₂ atmosphere.

Culture supernatants are collected for 8 days and Ig is measured by modification of specific isotype-selective ELISA assays as described by Marcelletti and Katz (as above). Modifications to the assay facilitate high efficiency. ELISA plates are prepared by coating overnight with DNP-KLH or DNP-OVA. After blocking with fetal bovine serum albumin (BSA), aliquots of each culture supernatant were diluted 1: 4 in phosphate buffered saline (PBS) containing BSA, sodium azide, and Tween 20 and added to the ELISA plate Incubate overnight in a box moistened at 4 ° C. IgE levels are quantified by continued incubation with biotinylated goat antimouse IgE (b-GAME), AP-streptavidin, and substrate.

Antigen-specific IgG1 is measured in a similar manner, except that the culture supernatant is diluted 200-fold and b-GAME is replaced with biotinylated goat anti-mouse IgG1 (b-GAMG1). Quantification of each isotype is determined by comparison with a standard curve. The detection level of all antibodies was about 200-400 pg / ml with less than 0.001% cross reactivity with other Ig isotypes in ELISA for IgE.

In vivo assay

Compounds found to be active in an ex vivo assay (above) are further tested for their activity in inhibiting IgE responses in their bodies. Mice irradiated with low doses of radiation prior to immunization with a carrier show an elevated IgE response to antigen challenge after 7 days. Test compounds are administered immediately before and after antigenic stimulation to determine the drug's ability to inhibit IgE responses. Antigen specific IgE, IgG1, and IgG2a levels in the serum are compared.

Female BALB / cByj mice were irradiated at 250 rad for 7 hours after the start of the daylight cycle. After 2 hours, mice are immunized by intravenous injection with 2 μg KLH in 4 mg of alum. After 6 days, drug injections are initiated once or twice daily for two to seven consecutive days. Intravenous injection and oral gavage are typically administered as a suspension (150 μl / injection) in saline containing 10% ethanol and 0.25% methylcellulose. Each treatment group consists of 5 to 6 mice. On day 2 of drug administration, 2 μg of DNP-KLH is administered by intravenous injection in 4 mg alum immediately after the drug injection in the morning. Mice are bled 7 days to 21 days after DNP-KLH challenge.

Antigen specific IgE, IgG1, and IgG2a antibodies are measured by ELISA. The ocular bleeding fluid is centrifuged at 14,000 rpm for 10 minutes to dilute the supernatant 5 times in saline and again centrifuged. Antibody concentrations in each bleeding blood are determined by ELISA (three pairs) of 4 dilutions and compared to standard curves: anti-DNP IgE (1: 100 to 1: 800), anti-DNP IgG2a (1: 100 to 1: 800), and anti-DNP IgG1 (1: 1600 to 1: 12800).

Active Compounds of the Invention

The following series of compounds belonging to class I, class II, and class III are found to be potent inhibitors of IgE in both in vivo and ex vivo models. These compounds also exhibit antiproliferative effects and can be used as such agents to treat hyperproliferative diseases, including cancer by themselves.

Compound of class I

One group of small molecule IgE inhibitors according to the invention comprises benzimidazole carboxamides defined by the following class (Class I):

In the above,

R is selected from the group consisting of H, C ₁ -C ₅ alkyl, benzyl, p-fluorobenzyl, and dialkylamino alkyl, and C ₁ -C ₅ alkyl is from the group consisting of linear, branched, or cyclic alkyl Selected;

R ₁ and R ₂ are independently H, alkyl, substituted alkyl, C ₃ -C ₉ cycloalkyl, substituted C ₃ -C ₉ cycloalkyl, polycyclic aliphatic groups, phenyl, substituted phenyl, naphthyl, substituted Selected from the group consisting of naphthyl, heteroaryl, and substituted heteroaryl, wherein the heteroaryl and substituted heteroaryl contain 1-3 heteroatoms, wherein the heteroatoms are independently composed of nitrogen, oxygen, and sulfur Selected from the group;

Substituted phenyl, substituted naphthyl, and substituted heteroaryl contain 1-3 substituents, wherein the substituents are H, halogen, polyhalogen, alkoxy group, substituted alkoxy, alkyl, substituted alkyl, dialkylamino Alkyl, hydroxyalkyl, OH, OCH ₃ , COOH, COOR ', COR', CN, CF ₃ , OCF ₃ , NO ₂ , NR'R ', NHCOR', and CONR'R ';

R ₃ and R ₄ are independently selected from the group consisting of H, alkyl, aryl, heteroaryl, and COR ′;

R 'is H, alkyl, substituted alkyl, C ₃ -C ₉ cycloalkyl, substituted C ₃ -C ₉ cycloalkyl, polycyclic aliphatic groups, phenyl, substituted phenyl, naphthyl, substituted naphthyl, heteroaryl , And substituted heteroaryl, wherein the heteroaryl and substituted heteroaryl contain 1-3 heteroatoms, and the heteroatoms are independently selected from the group consisting of nitrogen, oxygen, and sulfur;

X and Y are independently H, halogen, alkoxy, substituted alkoxy, alkyl, substituted alkyl, dialkylaminoalkyl, hydroxyalkyl, OH, OCH ₃ , COOH, CN, CF ₃ , OCF ₃ , NO ₂ , COOR '', CHO, and COR '';

R '' is C ₁ -C ₈ alkyl, wherein C ₁ -C ₈ alkyl is selected from the group consisting of linear, branched, or cyclic alkyl.

The following specific compounds are included within the definition of benzimidazole carboxamide class (Class I):

The compounds of class I can be synthesized by any conventional reaction known in the art. Examples of synthesis include the following reactions, designated as Synthesis Reaction I:

Synthesis of Compounds of Class I

Synthetic reaction I depicts one method that can be used to prepare compounds of class I. Those skilled in the art will appreciate that many different synthetic reactions can be used to synthesize the compounds of class I. In addition, those skilled in the art will understand that many other solvents, coupling agents, and reaction conditions may be used in the synthetic reactions to produce similar results.

In step 1, compound A or salts thereof are prepared from cyclocondensation of 3,4-diaminobenzoic acid or salts thereof and 4-nitrobenzaldehyde. Cyclocondensation can be prepared in a solvent by applying heat. One example of a solvent is nitrobenzene. The temperature of the cyclocondensation reaction is about 100 ° C to about 200 ° C, preferably about 155 ° C to about 160 ° C. The same compound reacts the diamine with p-nitrobenzoyl chloride in the presence of a base or other base such as triethylamine, DIEP, DMAP, or pyridine, and the resulting amide is PPA, H ₂ SO ₄ , or other It can be prepared by a two-step process, which cyclizes (by removing 1 mole of water) with a dehydrating agent to produce the benzimidazole ring.

In step 2, compound A or salts thereof was treated with ammonia or amine to give compound B or salts thereof. The amide formation reaction takes place in the presence of a coupling agent, or after conversion of compound A to acid chloride and in the presence of another base which absorbs the resulting acid, amines (eg aromatic amines, aliphatic amines, heterocyclics) in solvents. By amines). This can be done with or without heat. Examples of coupling agents are 1,1'-carbonyldiimidazole, EDC, and other similar coupling agents. Examples of solvents are N, N-dimethylformamide (DMF), TFT, pyridine, triethylamine, or mixed solvent systems such as DMF and THF.

In step 3, compound B or salts thereof may undergo a reduction reaction to produce compound C or salts thereof. The reduction reaction can be carried out by catalytic hydrogenation in the presence of a catalyst in a solvent system. Catalysts are Pd, Ni, Pt and the like. An example of a formulation used for catalytic hydrogenation is hydrogen in the presence of 5% Pd-C. The reduction reaction may occur in the presence of some acid in a solvent with hydroxyl groups, for example methanol or ethanol, or a mixed solvent system such as DMF-MeOH, or acetic acid, or in a solvent with hydroxyl groups.

In step 4, compound C or salts thereof are alkylated or acylated in amines by treatment with appropriate reagents. In synthetic reaction I, compound C was found to react with R ₃ -X and R ₄ -X to alkylate or acylate amines. It will be appreciated that R ₃ and R ₄ are groups that alkylate amines and X is a leaving group. Amino groups may be acylated with reagents such as acyl halides, anhydrides, carboxylic acids, carboxyl esters, or amides. The amino groups may be alkylated with alkyl alides in the presence of a base, preferably for the production of tertiary or hindered amines and the like. An alternative method of alkylating amino groups is reductive amination. In reductive amination reactions, amines condense with aldehydes or ketones to produce imines. Subsequently, the imine is reduced to produce an alkylated amine. In reductive amination reactions, the R ₃ and R ₄ groups may not have leaving groups upon reaction with amines. Another alternative method is the reaction of the diazo compound with the amine. The acidity of the amine is not large enough for the reaction to proceed without a catalyst, but BF ₃ , which converts the amine into a complex, can cause the reaction to occur. First copper cyanide can also be used as catalyst.

Compound D is a representative example of a compound of class I.

One skilled in the art will be able to make modifications to the sequence and to modify the appropriate reaction conditions from the shown analogous reaction or otherwise known analogous reactions, which can be used appropriately in the above procedure to prepare compounds of compounds A to D. There will be.

In the procedures described herein for the preparation of compounds A to D of the present invention, the need for protecting groups can be well recognized by those skilled in the art of organic chemistry, and therefore the use of appropriate protecting groups is not apparent for each group. It is necessarily included in the chemical reaction herein. Introduction and removal of such suitable protecting groups are well known in the art of organic chemistry. See, eg, T.W. Greene, "Protective Groups in Organic Synthesis", Wiley (New York), 1981.

The products of the reaction described herein are separated by conventional methods such as extraction, distillation, chromatography and the like.

Starting materials not described herein are commercially available, known, or can be prepared by methods known in the art.

Salts of the compounds A to D described above can be prepared by reacting the appropriate base or acid with the stoichiometric equivalents of the compounds of compounds A to D.

Compound of class II

Another group of small molecule IgE inhibitors according to the invention comprise benzimidazole-2-benzamides defined by the following class (Class II):

In the above,

R is selected from the group consisting of H, C ₁ -C ₅ alkyl, benzyl, p-fluorobenzyl, and dialkylamino alkyl, and C ₁ -C ₅ alkyl is from the group consisting of linear, branched, or cyclic alkyl Selected;

R ₁ and R ₂ are independently H, alkyl, substituted alkyl, C ₃ -C ₉ cycloalkyl, substituted C ₃ -C ₉ cycloalkyl, polycyclic aliphatic groups, phenyl, substituted phenyl, naphthyl, substituted Selected from the group consisting of naphthyl, heteroaryl, and substituted heteroaryl, wherein the heteroaryl and substituted heteroaryl contain 1-3 heteroatoms, wherein the heteroatoms are independently composed of nitrogen, oxygen, and sulfur Selected from the group;

Substituted phenyl, substituted naphthyl, and substituted heteroaryl contain 1-3 substituents, wherein the substituents are H, halogen, polyhalogen, alkoxy group, substituted alkoxy, alkyl, substituted alkyl, dialkylamino Alkyl, hydroxyalkyl, OH, OCH ₃ , COOH, COOR ', COR', CN, CF ₃ , OCF ₃ , NO ₂ , NR'R ', NHCOR', and CONR'R ';

R ₃ and R ₄ are independently selected from the group consisting of H, alkyl, aryl, heteroaryl, and COR ′;

R 'is H, alkyl, substituted alkyl, C ₃ -C ₉ cycloalkyl, substituted C ₃ -C ₉ cycloalkyl, polycyclic aliphatic groups, phenyl, substituted phenyl, naphthyl, substituted naphthyl, heteroaryl , And substituted heteroaryl, wherein the heteroaryl and substituted heteroaryl contain 1-3 heteroatoms, and the heteroatoms are independently selected from the group consisting of nitrogen, oxygen, and sulfur;

X and Y are independently H, halogen, alkoxy, substituted alkoxy, alkyl, substituted alkyl, dialkylaminoalkyl, hydroxyalkyl, OH, OCH ₃ , COOH, CN, CF ₃ , OCF ₃ , NO ₂ , COOR '', CHO, and COR '';

R '' is C ₁ -C ₈ alkyl, wherein C ₁ -C ₈ alkyl is selected from the group consisting of linear, branched, or cyclic alkyl.

The following specific compounds are included in the definition of class II:

Compounds of class II can be synthesized by conventional reactions in the art. Examples of synthesis include the following reactions designated Synthetic Reaction II:

Synthesis of Compounds of Class II

Synthetic reaction II illustrates one method that can be used to prepare compounds of class II. Those skilled in the art will appreciate that many different synthetic reactions can be used to synthesize the compounds of class I. In addition, those skilled in the art will understand that many other solvents, coupling agents, and reaction conditions may be used in the synthetic reactions to produce similar results.

In step 1, compound E or salts thereof are prepared from cyclocondensation of 4-nitro-1,2-phenylenediamine or salts thereof and alkyl (eg methyl 4-formylbenzoate). The cyclocondensation reaction can be carried out in a solvent by applying heat. Examples of the solvent include nitro benzene, or other solvent containing an oxidizing agent that converts imidazoline to imidazole. The same compound reacts the diamine with p-carboalkoxy benzoyl chloride in the presence of a base or other base such as triethylamine, DIEP, DMAP, or pyridine, and the resulting amide at PPA, H ₂ SO ₄ , Or by dehydrating (by removing 1 mole of water) with another dehydrating agent to produce a benzimidazole ring.

In step 2, the compound E or salts thereof can be treated with a base (for example lithium hydroxide solution or aqueous sodium hydroxide solution, etc.) which hydrolyzes the ester to an acid to give the chemical F or salts thereof. The deprotection reaction can take place in the presence of water or a solvent such as methanol or ethanol, THF or the like.

In step 3, compound F or salts thereof can be treated with ammonia or amines to give compound G or salts thereof. The amide formation reaction takes place in the presence of a coupling agent, or after conversion of compound A to acid chloride and in the presence of another base which absorbs the resulting acid, amines (eg aromatic amines, aliphatic amines, heterocyclics) in solvents. By amines). This can be done with or without heat. Examples of coupling agents include 1,1'-carbonyldiimidazole (CDI), EDC, and other similar coupling agents. Examples of the solvent include N, N-dimethylformamide (DMF), THF, pyridine, triethylamine, or mixed solvent systems such as DMF and THF.

In step 4, compound G or salts thereof may be subjected to reduction to yield compound H or salts thereof. The reduction reaction can be carried out by catalytic hydrogenation in the presence of a catalyst in a solvent system. Catalysts are Pd, Ni, Pt and the like. An example of a formulation used for catalytic hydrogenation is hydrogen in the presence of 5% Pd-C. The reduction reaction may occur in the presence of some acid in a solvent with hydroxyl groups, for example methanol or ethanol, or a mixed solvent system such as DMF-MeOH, or acetic acid, or in a solvent with hydroxyl groups.

In step 5, the compound is H or salts thereof are treated with acyl halides to afford formula I or salts thereof. The acylation reaction can occur in a solvent such as THF, DMF or Et3N, pyridine and the like in the presence of a base such as triethylamine, DIEF, DMAP or pyridine. The reaction can occur with or without heating. One specific example of a base is pyridine. One specific example of a solvent is tetrahydrofuran (THF).

If necessary, in step 6, compound I or a salt thereof is treated with an alkyl halide in the presence of a base to effect N-alkylation of the amide. Secondary amines can be alkylated with a base such as sodium hydride for proton removal and then reacted with an alkyl halide. This reaction can be carried out under conventional solvent systems or phase transfer conditions. In another method, N-alkyl amides can also be prepared by starting the alcohol from the alcohol at room temperature in equal molar amounts of amide, Ph ₃ P, and diethyl azodicarboxylate (EtOOCN = NCOOEt).

Compound I (b) is a representative example of a compound of class II.

One skilled in the art will be able to make modifications to the sequence and to modify the appropriate reaction conditions from the shown analogous reaction or otherwise known analogous reactions, which can be used appropriately in the above procedure to prepare compounds of compound EI (b). Could be.

In the process described herein for the preparation of compound EI (b) of the present invention, the need for protecting groups can be well recognized by one skilled in the art of organic chemistry, so the use of appropriate protecting groups is not explicitly described for each group. However, it is necessarily included in the chemical reaction of the present application. Introduction and removal of such suitable protecting groups are well known in the art of organic chemistry. See, eg, T.W. Greene, "Protective Groups in Organic Synthesis", Wiley (New York), 1981.

The products of the reaction described herein are separated by conventional methods such as extraction, distillation, chromatography and the like.

Starting materials not described herein are commercially available, known, or can be prepared by methods known in the art.

Salts of the compounds E-I (b) described above can be prepared by reaction of the appropriate base or acid with the stoichiometric equivalents of the compounds of compounds E-I (b).

Compound of class III

Another group of small molecule IgE inhibitors according to the invention comprise benzimidazole-bis-carboxamides defined by the following class (Class III):

In the above,

R is selected from the group consisting of H, C ₁ -C ₅ alkyl, benzyl, p-fluorobenzyl, and dialkylamino alkyl, and C ₁ -C ₅ alkyl is from the group consisting of linear, branched, or cyclic alkyl Selected;

R ₁ and R ₂ are independently H, alkyl, substituted alkyl, C ₃ -C ₉ cycloalkyl, substituted C ₃ -C ₉ cycloalkyl, polycyclic aliphatic groups, phenyl, substituted phenyl, naphthyl, substituted Selected from the group consisting of naphthyl, heteroaryl, and substituted heteroaryl, wherein the heteroaryl and substituted heteroaryl contain 1-3 heteroatoms, wherein the heteroatoms are independently composed of nitrogen, oxygen, and sulfur Selected from the group;

Substituted phenyl, substituted naphthyl, and substituted heteroaryl contain 1-3 substituents, wherein the substituents are H, halogen, polyhalogen, alkoxy group, substituted alkoxy, alkyl, substituted alkyl, dialkylamino Alkyl, hydroxyalkyl, OH, OCH ₃ , COOH, COOR ', COR', CN, CF ₃ , OCF ₃ , NO ₂ , NR'R ', NHCOR', and CONR'R ';

R ₃ and R ₄ are independently selected from the group consisting of H, alkyl, aryl, heteroaryl, and COR ′;

R 'is H, alkyl, substituted alkyl, C ₃ -C ₉ cycloalkyl, substituted C ₃ -C ₉ cycloalkyl, polycyclic aliphatic groups, phenyl, substituted phenyl, naphthyl, substituted naphthyl, heteroaryl , And substituted heteroaryl, wherein the heteroaryl and substituted heteroaryl contain 1-3 heteroatoms, and the heteroatoms are independently selected from the group consisting of nitrogen, oxygen, and sulfur;

X and Y are independently H, halogen, alkoxy, substituted alkoxy, alkyl, substituted alkyl, dialkylaminoalkyl, hydroxyalkyl, OH, OCH ₃ , COOH, CN, CF ₃ , OCF ₃ , NO ₂ , COOR '', CHO, and COR '';

R '' is C ₁ -C ₈ alkyl, wherein C ₁ -C ₈ alkyl is selected from the group consisting of linear, branched, or cyclic alkyl.

The following specific compounds are included in the definition of class III:

Compounds of class III may be synthesized by conventional reactions known in the art. Examples of the synthesis include the following reactions designated as synthesis reaction III:

Synthesis of Compounds of Class III

Synthesis reaction 3 illustrates one method that can be used to prepare compounds of class III. Those skilled in the art will appreciate that many different synthetic reactions can be used to synthesize Class III compounds. In addition, those skilled in the art will understand that many different solvents, coupling agents, and reaction conditions can be used in the synthetic reactions to achieve similar results.

In step I, compound J can be prepared from cyclocondensation of 3,4-diaminobenzoic acid or salts thereof and 4-alkoxycarbonyl benzaldehyde. The cyclocondensation reaction can be carried out in a solvent by applying heat. Examples of solvents are other solvents that can convert imidazolines to imidazoles together with nitrobenzene and oxidants. The same compound reacts the diamine with p-carboalkoxy benzoyl chloride in the presence of a base such as triethylamine, DIEP, DMAP, or pyridine or another base, and the resulting amide at PPA, H ₂ SO ₄ , Or by dehydrating one mole of water with another dehydrating agent to produce a benzimidazole ring.

In step 2, compound J or a salt thereof is treated with an inorganic acid halide such as thionyl chloride, POCl ₃ , PCl ₅ , or the like, or an organic acid chloride such as oxalyl chloride, or a mixed anhydride such as t-butyl chloroformate to give compound K or Similar reaction intermediates and salts thereof are obtained. The reaction can take place in a solvent in the presence of an inorganic acid halide preparation or a chlorinated organic acid or mixed anhydride. One specific example of an inorganic acid halide formulation is thionyl chloride. One example of a solvent is DMF.

In step 3, compound K or salts thereof are treated with ammonia or amines to yield compound L or salts thereof. The amide formation reaction is carried out in the presence of a coupling agent or in the solvent in the presence of another base which converts the compound K or salts thereof into chloride acid or mixed anhydride and then absorbs the resulting acid. By reacting with amines such as cyclic amines and the like. This can be done with or without heat. Examples of coupling agents are 1,1'-carbonyldiimidazole (CDI), EDC, and other similar coupling agents. Amide forming reactions can occur in the presence of a base in a solvent. Examples of solvents include solvent systems such as N, N-dimethylformamide (DMF), THF, pyridine, triethylamine, or DNF and THF and the like.

In step 4, compound L or a salt thereof is treated with a base to hydrolyze the ester to an acid using a base such as a lithium hydroxide solution or a sodium hydroxide aqueous solution or the like to give compound M or a salt thereof. The deprotection reaction can occur in the presence of a solvent such as water or alcohol, including methanol, ethanol, THF and the like.

In step 5, compound M or salts thereof is treated with ammonia or amine to yield compound O or salts thereof. The amine forming reaction is carried out in the presence of a coupling agent, or in the solvent in the presence of another base which converts the compound K or its salts into chloride acids or mixed anhydrides, and then absorbs the resulting acids. By reacting with amines such as cyclic amines and the like. This can be done with or without heat. Examples of coupling agents are 1,1'-carbonyldiimidazole (CDI), EDC, and other similar coupling agents. Amide forming reactions can occur in the presence of a base in a solvent. Examples of solvents include solvent systems such as N, N-dimethylformamide (DMF), THF, pyridine, triethylamine, or DNF and THF and the like.

Alternatively, compound M or salts thereof are treated with inorganic acid halides to yield compound N or salts thereof. The reaction is obtained in a solvent in the presence of an inorganic acid halide formulation. One example of an inorganic acid halide formulation is thionyl chloride. An example of a solvent is DMF.

Compound N or a salt thereof is then treated with ammonia or an amine to yield compound O or a salt thereof. The amide formation reaction can take place in a solvent in the presence of a base.

Compound O is a representative example of class III.

One skilled in the art can recognize modifications in the sequence and can also add modifications to the appropriate reaction conditions from similar or otherwise known methods which may be suitably used in the above methods for preparing compounds of compounds J to O. There will be.

In the methods described herein for the preparation of compounds J to O of the present invention, the need for protecting groups will generally be well recognized by one skilled in the art of organic chemistry and therefore the use of suitable protecting groups is not clearly exemplified by such groups. It will necessarily be included in the reaction process herein. Introduction and removal of such appropriate protecting groups are well known in the field of organic chemistry and are described, for example, in T.W. Green, "Protective Groups in Organic Synthesis", Wiley (New York), 1981.

The products of the reaction described herein can be separated by conventional means such as extraction, distillation, chromatography and the like.

Starting materials not described herein may be commercially available, known, or prepared by methods known in the art.

Salts of the compounds J to O described above are prepared by reacting the appropriate base or acid with the stoichiometric equivalents of the compounds of compounds J to O.

Example 1

Preparation of Compounds of Class 1

A. Preparation of Benzimidazole Carboxamides

Preparation of 2- [4-nitrophenyl] benzimidazole-5-carboxylic acid:

A mixture of 3,4-diamino benzoic acid (300 g; 1.97 mol) and p-nitro benzaldehyde (298 g; 1.97 mol) in nitrobenzene (15 L) was heated to about 155 ° C. to 160 ° C. overnight. The reaction mixture was cooled to room temperature and the precipitated solid was filtered off and washed several times with ether to remove all nitrobenzene. The product was treated with charcoal in hot DMF (2 L), filtered and then stirred at room temperature and diluted with ether (6 L) to give 393 g of solid. The crude solid was again treated with charcoal in hot DMF (1 L), filtered and then diluted with methanol (5 L) and cooled to about 0 ° C. The product was then crystallized from DMF and ether to give 225 g of pure product. This was used in the next step.

Preparation of 2- [4-nitrophenyl] benzimidazole-5- (N-cyclohexyl) carboxylic amide:

The carboxylic acid was then converted to cyclohexyl amide as follows. A mixture of acid (1.0 g, 3.53 mmol) and CDI (0.6 g, 4.24 mmol) in DMF (20 mL) was stirred at room temperature for 3 hours, then cooled to 0 ° C. and cyclohexyl amine (0.36 g, 0.42 mL) , 3.65 mmol), and the mixture was stirred for 1 hour and then filtered. The crude product was recrystallized from DMF and ether to give 0.68 g of the desired product. It was used in the next step without any further purification. The product appeared as a single point on TLC, different from the starting material.

2- [4-amino-phenyl] benzimidazole-5- (N-cyclohexyl) carboxylic amide:

The mixture of carboxy amide (0.5 g, 1.37 mmol), 5% Pd-C (0.35 g), and methanol was stirred under hydrogen gas atmosphere until the required amount of hydrogen was absorbed. The catalyst was filtered off and the filtrate was concentrated to give a solid (430 g). The product showed a different TLC single point from the starting material.

2- [4-((5-methyl isoxazolyl) -3-carbamido) phenyl] benzimidazole-5- (N-cyclohexyl) carboxylic amide:

A mixture of 5-methyl isoxazole-3-carboxylic acid (0.23 g, 1.80 mmol), oxalyl chloride (0.46 g, 3.58 mmol), and a drop of DMF in CH ₂ Cl ₂ (10 mL) was heated for 1 hour. Reflux for a while. The reaction mixture was concentrated to dryness and crude acid chloride was used as in the next reaction. The mixture of amine (0.5 g, 1.50 mmol), crude acid chloride, THF (50 mL), and pyridine (0.54 g) was refluxed overnight. The reaction mixture was poured into water (600 mL). The crude product was filtered off, washed with water and hexanes and dried. The yield was 380 mg and the melting point was> 310 ° C. The product showed a single point on TLC.

B. 2- [4- (N- (1-adamantyl-carboxamido) phenyl] benzimidazole-5- (N-2-pyridyl) carboxy amide

Preparation of 2- [4-nitrophenyl] benzimidazole-5- (N-2-pyridyl) carboxylic amide:

A mixture of 2- [4-nitrophenyl] benzimidazole-5-carboxylic acid (20.0 g, 0.071 mol) and CDI (17.2 g, 0.11 mol) in DMF was stirred at room temperature for 3 hours and then cooled to 0 ° C. After 2-amino pyridine (7.3 g, 0.078 mol) was added, stirring was continued at room temperature overnight. HPLC results showed that the reaction was incomplete. Additional CDI (17.2 g) and 2-aminopyridine (7.3 g) were added and then heated to form a clear solution and stirred for an additional 24 hours. TLC analysis showed the reaction was incomplete, so an additional amount of 2-aminopyridine (7.3 g) was added with DMAP (13.4 g, 0.11 mol) and stirred overnight. The PLC results showed the reaction was complete and the reaction mixture was poured into water (3.0 L), stirred for 1 hour, filtered, washed with water and ether (3 x 100 mL) and dried. The yield was 17 g (68%). It was used in the next step without further purification. TLC showed a single point (CH ₂ Cl ₂ -CH ₃ OH: 9: 1).

Preparation of 2- [4-amino-phenyl] benzimidazole-5- (N-2-pyridyl) carboxylic amide:

2- [4-nitrophenyl] benzimidazole-5- (N-2-pyridyl) carboxy amide (17 g, 0.47 mol), 5% Pd-C (93.0 g) in methanol (1.0 L), and DMF (200 mL) was stirred under hydrogen until the reaction was complete. The catalyst was filtered, concentrated and poured into water (5.0 L), filtered, washed with water and ether (3 x 100 mL) and oven dried in vacuo at 80 ° C. The yield was 10 g and the melting point was 260 ° C to 265 ° C. TLC results showed a single point using CH ₂ C ₁₂ -CH ₃ OH (9: 1) as eluent.

2- [4- (N- (1-adamantyl-carboxamido) phenyl] benzimidazole-5- (N-2-pyridyl) carboxylic amide:

A mixture of 2- [4-amido-phenyl] benzimidazole-5- (N-2-pyridyl) carboxylic amide (2.6 g, 0.7.89 mmol) and pyridine (2.9 mL) in THF (250 mL) Heat to produce a clear solution. The reaction mixture was then treated with 1-adamantylcarbonyl chloride (1.88 g, 9.47 mmol) in THF (10 mL) and the mixture was heated to reflux for 24 h. The mixture was poured into water (1.5 L) and stirred for 1 hour, filtered, washed with water (3 x 50 mL) and ether (3 x 50 mL) and dried. It was treated with charcoal and recrystallized from THF and methanol. The filtrate was diluted with ether (150 mL) and cooled to -70 ° C for 4 h at which time the product crystallized. It was filtered, washed with ether and dried. The yield was 2.9 g and the melting point was 333 ° C to 336 ° C. TLC results showed a single point using CH ₂ Cl ₂ -CH ₃ OH (9: 1) as eluent.

Example 2

Preparation of Compounds of Class II

A. Preparation of 2- (4-N- (2-methyl-cyclohexyl) benzamido) -5- (benzamido) benzimidazole:

Preparation of 2- (4-carbomethoxy-phenyl) -5-nitro-benzimidazole:

A mixture of 4-nitrophenylene-1,2-diamine (634 g) and methyl-4-formyl benzoate (680 g) is heated in nitrobenzene (17 L) to 150-155 ° C for 24 hours, cooled to room temperature The product was filtered off, washed with ether (3 × 1.0 L) and dried to afford the desired product. The yield is 800 g and TLC showed a single point: using CH ₂ Cl—CH ₃ OH of (9: 1).

Preparation of 2- (4-carboxy-phenyl) -5-nitro-benzimidazole:

The mixture of ester (800 g), THF (2.7 L), and water (2.6 L) was treated with LiOH (339 g) and stirred at room temperature. The progress of the reaction was followed by TLC until the hydrolysis was complete. The reaction mixture was diluted with hot water (2.0 L), charcoal and then filtered. The filtrate was diluted with 2.0 kg ice and water (2.0 L) and acidified with concentrated HCl. The product was filtered, washed with water and then diluted with hot DMF (7.0 L) and refrigerated to 4.0 ° C. The product was filtered off, washed with ether and dried. The yield was 537 g, the melting point was above 355 ° C., and the TLC results showed a single point: using CH ₂ Cl—CH ₃ OH of (9: 1).

Preparation of 2- (4-N- (2-methyl-cyclohexyl) benzamido) -5-nitro-benzimidazole:

The mixture of said acid (20.0 g) in DMF (400 mL) was treated with CDI (13.7 g) and the mixture was stirred at room temperature for 2.0 h and then with 2-methyl-cyclohexyl amine (11.2 g). The reaction mixture was heated to reflux for 16 hours. It was poured into ice-water (3.0 L) and stirred for 16 hours at room temperature. The crude product was filtered off, washed with water (3 × 100 mL) and ether and dried. Yield was 19 g and TLC showed a single point: (9: 1) -CH ₂ Cl ₂ -CH ₃ OH.

Preparation of 2- (4-N- (2-methyl-cyclohexyl) benzamido) -5-amino-benzimidazole:

The nitro amide (19.0 g) was hydrogenated in the presence of 5% Pd-C (4.0 g) in MeOH (600 mL). The catalyst was filtered off and the filtrate was concentrated in vacuo to treat the residue with ether (200 mL) and filtered. The residue was then recrystallized from THF, MeOH, and hexanes. The product was filtered off, washed with hexanes and oven dried. Yield was 10.5 g and TLC showed a single point: (9: 1) -CH ₂ Cl ₂ -CH ₃ OH.

Preparation of 2- (4-N- (2-methyl-cyclohexyl) benzamido) -5- (benzamido) -benzimidazole:

A mixture of this amine (0.5 g) in THF (50 mL) and pyridine (0.53 mL) was heated to a clear solution and the solution was treated in small portions with benzoyl chloride (0.24 g) in 10 mL THF. The reaction mixture was heated to reflux for 24 hours, cooled and then poured into water (600 mL). The product was filtered off, washed with water and ether and dried. The crude product was treated with charcoal in hot THF-methanol and filtered. The filtrate was diluted with ether and refrigerated. The product was filtered off, washed with ether and dried. Yield 338 mg, melting point 285-289 ° C., TLC showed single point: (9: 1) -CH ₂ Cl ₂ -CH ₃ OH.

Example 3

Preparation of Compounds of Class 3

A. Preparation of 2- (4-cyclohexylcarbamoyl-phenyl) -3H-benzoimidazole-5-carboxylic acid cyclohexamide :

Preparation of 2- (4-methoxycarbonyl-phenyl) -3H-benzoimidazole-5-carboxylic acid:

A mixture of 3,4-diaminobenzoic acid (300 g) and methyl-4-formyl benzoate (324 g) in nitrobenzene (8.0 L) was heated at 150 to 155 ° C. for 24 hours and then cooled to <10 ° C. At this time, the product crystallized. After filtration it was washed with ether (3 x 200 mL) and dried in vacuo. The yield was 320g and the melting point was 308-309 degreeC. It was used in the next step without further purification.

Preparation of 2- (4-carboxy-phenyl) -3H-benzoimidazole-5-carboxylic acid:

A mixture of 2- (4-methoxycarbonyl-phenyl) -3H-benzoimidazole-5-carboxylic acid (4.1 g) and LiOH (1.74 g) in TMF (15 mL) and water (18 mL) was cooled to room temperature. After stirring for 16 h in hot, it was mixed with hot water (100 mL) and charcoal. The mixture was filtered, the filtrate was diluted with ice and water (100 mL) and acidified with concentrated HCl. The crude dibasic acid was filtered off, washed with water and ether and then recrystallized from THF and MeOH. The yield was 3.3 g and TLC showed a sweet point [CH ₂ Cl ₂ -CH ₃ OH (9: 1)].

Preparation of 2- (4-cyclohexylcarbamoyl-phenyl) -3H-benzoimidazole-5-carboxylic acid cyclohexamide:

A mixture of 2- (4-carboxy-phenyl) -3H-benzoimidazole-5-carboxylic acid (0.50 g) and N-methylmorpholine (0.72 g) in DMF (50 mL) was -10 ° C to 20 ° C. After refrigeration with isobutyl chloroformate (0.60 g) was added in small portions while maintaining the temperature. After 10 minutes, cyclohexylamine (0.53 g) in DMF (10 mL) was added in small portions. The mixture was then slowly brought to room temperature and stirred for 48 hours. TLC was performed after completion of the reaction. The mixture was then poured into water (600 mL), filtered, washed with water and ether and recrystallized from THF and MeOH. Second recrystallization was carried out using charcoal to give the pure product. The yield was 270 mg and the melting point was 334 to 337 ° C.

B. Preparation of 2- (4-carbomethoxy-phenyl) -5- (N-cyclohexyl-carboxamido) -benzimidazole :

A mixture of 2- (4-methoxycarbonyl-phenyl) -3H-benzoimidazole-5-carboxylic acid (5.0 g) and CDI (7.1 g) in DMF (250 mL) was stirred for 3 hours at room temperature , Cyclohexyl amine (2.0 g) and refluxed for 96 hours. After cooling the reaction mixture was poured into water (2.0 L) and stirred for 16 h at room temperature. The product was filtered off, washed with water and ether and then recrystallized from THF, methanol, and ether. The yield was 0.4 g and TLC showed a single point [(9: 2) -CH ₂ Cl ₂ -CH ₃ OH).

This ester was hydrolyzed to yield an acid that was coupled with various amines to produce asymmetric bis amides.

C. Preparation of 2- (4-N-cyclohexyl) -benzamido) -benzimidazole-5-carboxylic acid and asymmetric bisamide :

A mixture of 3,4-diaminobenzosa (1.71 g) and 4- (N-cyclohexyl) -benzamido) -benzaldehyde (2.6 g) in 200 ml nitrobenzene was heated to 150-155 ° C. for 16 hours. I was. It was cooled, filtered, washed with ether and dried. Yield was 1.9 g and TLC showed a single point [(9: 1) -CH ₂ Cl ₂ -CH ₃ OH].

The acid was then coupled with various amines using CDI in DMF and / or THF to give an unsymmetrical bis amide.

D. Preparation of 2- (4-carboxy-phenyl) -5- (N-cyclohexyl-carboxamido) -benzimidazole

LiOH (1.73 g) to a mixture of 5.2 g of 2- (4-carbomethoxy-phenyl) -5- (N-cyclohexyl-carboxamido) -benzimidazole in THF (50 mL) and water (40 mL) ) Was added. The mixture was stirred at room temperature for 2 hours, then mixed with charcoal, stirred with some heating and filtered. The filtrate was refrigerated in ice and acidified to pH 1.0 with concentrated HCl. The acid was filtered off, washed with water and ether and dried. The yield was 4.7 g, 95% by HPLC, and the melting point was 311 to 314 ° C. This was coupled with various amines using CDI or isobutyl chloroformate as the coupling agent to give asymmetric bis amides. One such example is as follows: a mixture of acid (0.5 g) and N-methyl morpholine (0.64 g) in THF (50 mL) is cooled to -10 to 20 ° C., followed by isobutyl chloroformate (0.3 g). Small amount). After the reaction mixture was stirred for 10 minutes, 1-adamantane amine was added to the mixture and the reaction mixture was stirred for 15 hours. The reaction mixture was poured into water and ice, filtered and washed with water, ether and then recrystallized from THF / MeOH to give the desired product. The yield was 320 mg and the melting point was> 355 ° C.

Example 4

Inhibition of IgE Response

Inhibitory activity of small molecules of the invention was assayed using both in vitro and in vivo assays as described above. All of the compounds presented above have the activity of inhibiting IgE. In in vitro assays, compounds of class I-III showed 50% inhibition at concentrations ranging from 1 pM to 100 μM. In an in vivo assay, the compound was effective at a concentration of less than about 0.01 mg / kg / day to about 100 mg / kg / day, at which time it was divided into consecutive divided doses (eg, per day) for at least 2 to 7 days. 2-4 times). As such, the small molecule inhibitors of the present invention are disclosed to be useful in lowering antigen-induced increases in IgE concentrations and consequently to be useful in the treatment of IgE dependent processes such as general allergies, and especially allergic asthma.

Example 5

Effect on Cell Proliferation

Various experiments were conducted to determine the effect of benzimidazole compounds on cell proliferation. This experiment measured ³ H-thymidine incorporation in ultimately proliferating cellular DNA. The specific process differed depending on the cells and stimulation. Cells derived from the mouse spleen were incubated at 3 million per ml and the cell lines seeded at 10 million to 1 million per ml. Spleen B cells are isolated by T cell bleeding and treated with phorbol myristate acetate (PMA) (10 ng / ml) and inomycin (100 nM), or IL-4 (10 ng / ml) and anti CD40 Ab (100 ng / ml). Stimulated. T cells were first incubated with a cocktail of anti-Thy1 activity (10%), anti-CD4 Ab (0.5 μg / ml), and anti-CD8 Ab (0.5 μg / ml) followed by guinea pig complement (adsorption). Bleeding before incubation by incubation. Cell lines were stimulated or not stimulated with human epidermal growth factor (EGF) (100 ng / ml). All cells were incubated in 96-well plates for 2-3 days and pulsed with 50 μl of 3H-thymidine (50 μCi / ml) for 6-14 hours.

In splenocytes, compound I.82 is a B cell proliferative response to PMA / inomycin and IL-4 / anti-CD40Ab with approximately the same potency as inhibiting the IgE response to IL-4 / anti-CD40 Ab in vitro. (FIG. 1) was inhibited. Similar inhibitory effects on Compound I.82 were obtained in ConA-stimulated T cell proliferation and LPS-stimulated B cell proliferation (performed by MDS Pharma), suggesting a lack of specificity in the action of these drugs. do. On the other hand, the battery of immunological tests performed with compound I.82 hardly demonstrated an effect other than inhibition of ConA-stimulated cytokine release.

In tumor cells, the use of splenic lymphocytes led to further analysis of cell proliferation by measuring tumor cell proliferation in the presence of such drugs. Initial analysis was performed using M12.2.1 lymphoma cells stimulated or not stimulated with IL-4 / anti-CD40 Ab. Compound I.82 inhibited the proliferation of M12.4.1 cells except for the low efficacy observed in stimulated spleen cells. However, the efficacy of compound I.82 was increased when cells were cultured with IL-4 / antiCD40Ab. Such stimulation is known to induce the activity of NF- _k B in M12.3.1 cells.

Similar methods were established to establish the selectivity of antiproliferative activity by testing batteries of tumor cell lines derived from various tissues, primarily various tissues of human origin. Attempts were made to generate proliferation data from two or more cell lines of each tissue selected. Only a few cell lines were inhibited by less than 100 nM of each compound and most of the remaining cells needed higher concentrations. Because of previous Western blot results with known properties and compounds of some of the cell lines tested, there is evidence to suggest a link between NF- _k B inhibition and the action of the drug. Breast cancer cells provide an excellent model for testing this phenomenon because they are mainly two types of estrogen receptor (ER) -positive and ER-negative. ER-negative cells have the property to differentiate less, have higher density of EGF receptor expression, and are easier to handle. Proliferation of ER-negative / EGFR-positive cells also tends to be driven by NF- _k B, so the selection of these cells was tested for proliferative response to the drug in vitro. Proliferation of all EFG-reactive cell lines was effectively inhibited by Compound I.82 in vitro. In contrast, only two ER-positive cell lines were potently inhibited by the drug.

Compound I.82 has antiproliferative activity on T and B lymphocytes exposed to various immune stimuli in vitro. This action is very effective and comparable to their IgE inhibitory activity. Although the mechanism of this action is not known, much is known about the mechanism of IL-4 / anti-CD40 Ab-induced IgE production. The main factor in this reaction is the transcriptional activator, NF- _k B. These factors are associated with the proliferation of many tumor cells and therefore these drugs were tested for activity against the proliferation of various tumor cell lines in vitro. Our experiments revealed that many tumor cell lines are sensitive to the effects of compound I.82, and that the proliferation of many of these sensitive cell lines can be driven by the NF- _k B factor. However, other cell lines are known to be operated by factors other than NF- _k B (eg, ER-positive HCC 1500 and ZR-75-1). Thus, compound I.82 appears to selectively act on certain tumor cells. Other compounds disclosed according to the invention, in particular those compounds which are structurally similar to compound I.82, are expected to exhibit similar properties.

Treatment plan

The amount of benzimidazole compound that may be effective in treating a particular allergy or used as an antiproliferative agent will vary depending on the nature of the disease and can be determined by standard clinical techniques. The exact amount to be applied in a given situation will depend on the severity of the compound and condition and should be determined at the discretion of the situation of the user and each patient.

As an antiallergic treatment, the appropriate dose can be determined and adjusted by the user based on the dose response relationship between the IgE of the patient as well as the standard signs of pulmonary changes and hemodynamic changes. Moreover, those skilled in the art will appreciate that dosage ranges may be in accordance with the protocols disclosed herein for in vitro and in vivo screening (see Hasegawa et al., J. Med. Chem. 40: 395-407, 1997; and Ohmori et al., Int. J. Immunopharmacol. 15: 573-579, 1993); employing similar ex vivo and in vivo assays for determining dise response relationships for IgE suppression by naphthalene derivatives; It will be appreciated that it may be determined without further experimentation, incorporated herein by reference.

Initially, for an antiallergic or anti-asthmatic effect, a suitable dose of the compound will generally range from about 0.001 mg to about 300 mg / kg body weight / day in divided doses, preferably from about 0.01 mg to divided doses 100 mg / kg body weight / day. The compound is preferably administered systemically as a pharmaceutical formulation suitable for routes such as oral, nebulization, intravenous, subcutaneous, or by other routes that may be effective in providing a conventional administration of the active compound. Compositions of pharmaceutical formulations are known in the art. The method of treatment preferably includes periodic administration. Moreover, sustained treatment may be indicated when the allergic response appears to be caused by continued exposure to allergens. Daily or two daily administrations were effective in inhibiting the IgE response to single antigen challenge in animals when performed continuously for two to seven days in a row. Thus, in a preferred embodiment, the compound is administered two or more consecutive days at regular intervals. However, treatment regimens, including dosing frequency and duration of treatment, may be determined by one skilled in the art and may be modified as necessary to provide optimal control of IgE lowering depending on the nature of the allergen, dose, frequency, duration of allergen exposure, and standard clinical signs. Can be.

In one embodiment of the invention, the IgE-inhibiting compound may be administered with one or more other small molecule inhibitors, disclosed for optimally controlling the drop in the IgE response of the patient. In addition, one or more compounds of the present invention may be administered in combination with other drugs that are later developed or already known for the treatment of the underlying cause as well as the acute symptoms of allergy or asthma. Such combination treatments within the scope of the present invention include mixing one or more small molecule IgE inhibitors with one or more additional ingredients known to be effective in reducing one or more symptoms of a disease state. In one variation, the small molecule IgE-inhibitors disclosed herein may be administered separately from the additional drug during the same course of the disease state, wherein both IgE-inhibitor and emollient compounds will be administered according to their respective effective methods of treatment. Can be.

As antiproliferative treatment, appropriate doses of the benzimidazole compounds disclosed herein can be determined by one skilled in the art. Pharmacologists and oncologists can easily determine the appropriate dose required for each individual patient without further experimentation based on standard treatment techniques used for other antiproliferative and chemotherapeutic agents.

Initially, suitable doses of the antiproliferative benzimidazole compounds range from about 0.001 mg to about 300 mg / kg body weight / day in divided doses, preferably from about 0.01 mg to 100 mg / kg body weight / day in divided doses. It will be in the range of. Most preferably, for an anticancer effect, the dose will range from about 1 mg to 100 mg / kg body weight / day. The compound is preferably administered systemically in a pharmaceutical formulation suitable for routes such as oral, nebulization, intravenous, subcutaneous or by other routes that may be effective for systemic administration of the active compound.

Ideally, one or more benzimidazole compounds of the present invention should be administered to achieve the maximum plasma concentration of the active agent as determined by one skilled in the art. In order to achieve an appropriate shape level, the pharmaceutical formulations may be injected intravenously in a suitable solution such as saline solution or administered as a bolus of the active ingredient.

The method of treatment according to some embodiments of the invention preferably comprises periodic administration. Moreover, as with other chemotherapeutic agents, sustained therapy may be indicated. Weekly, daily or twice daily administration for a period of 1 to 3 years may be required for some patients. Thus, in a preferred embodiment, the compound is administered for at least 6 months at regular intervals. However, treatment methods, including frequency of administration and duration of treatment, can be determined by one skilled in the art and provide optimal antiproliferative effects depending on the nature of the disease, the extent of abnormal cell proliferation, the type of cancer, the affected tissue, and standard clinical symptoms. It may be modified if necessary to make.

Those skilled in the art will appreciate that the ideal concentration of antiproliferative compound in the formulation will depend on other known factors as well as pharmacokinetic parameters such as adsorption, inactivation, metabolism and clearance rate of the drug. Those skilled in the art will also recognize that concentrations vary with the severity of the condition to be treated. Other factors that may affect the therapeutic dose include tumor location, age and sex of the patient, other illnesses, and prior exposure to other drugs. Those skilled in the art will appreciate that for a particular patient a particular treatment method may be evaluated and adjusted over time according to the needs of the individual patient and the professional judgment of the practitioner administering the treatment.

In one preferred embodiment, the compound of the present invention is administered orally. Preferably, the oral formulation will comprise an inert diluent or edible single substance. Oral doses may be encapsulated in gelatin or formed into tablets. Oral administration can also be carried out by using granules, grains or labor, syrups, suspensions or solutions. Those skilled in the art will appreciate that many acceptable oral compositions can be used in accordance with some embodiments of the present invention. For example, the active compound may be combined with standard excipients, adjuvants, lubricants, sweeteners, enteric coatings, buffers, stabilizers and the like.

In one embodiment of the invention, the compounds of the invention may be modified such that the active compound comprises a target moiety that targets or concentrates the compound at the active site. Target residues include, but are not limited to, antibodies, antigen fragments or derivatives, cytokines, and receptor ligands expressed in the cells to be treated.

In some embodiments, the compounds of the present invention are administered in combination with other active agents that enhance or facilitate the action of the benzimidazole compound or cause other independent improving effects. These additional active agents include, but are not limited to, antifungal agents, antiviral agents, antibiotics, anti-inflammatory agents, and anti-agents. Protective agents can also be used, including carriers or agents that protect the active benzimidazole compounds from rapid metabolism, degradation, or elimination. Controlled release formulations may also be used in accordance with some embodiments of the present invention.

In one embodiment of the invention, the one or more antiproliferative compounds may be administered with one or more anticancer agents or therapeutic agents that produce an optimal antiproliferative effect. The anticancer agent is an alkylating agent (Romustine, Carmustine, Streptozosin, Mechloretamine, Melphalan, Urasyl Nitrogen Mustard, Chlorambucil Cyclophosphamide, Iphosphamide, Cisplatin, Carboplatin Mitomycin Thiotepa Dacarbazine Procarbazine, hexamethyl melamine, triethylene melamine, busulfan, fifobroman, and mitotan), metabolic agents (methotrexate, trimetrexate pentostatin, cystravin, ara-CMP, fludarabine phosphate, hydride Oxyurea, fluorouracil, fluorouridine, chlorodeoxyadenosine, gemcitabine, thioguanine, and 6-mercaptopurine); DN cutters (bleomycin); Topoisomerase I activity inhibitors (topotecan irinotecan and camptothecin); Topoisomerase II activity inhibitors (daunorubicin, doxorubicin, idarubicin, mitosantron, teniposide, and etoposide); DNA binding agents (dactinomycin and mitramycin); And spindle inhibitors (vinblastine, vincristine, nabelvin, paclitaxel and docetaxel).

In addition, one or more compounds of the present invention may be administered in combination with other therapies such as radiation therapy, immunotherapy, gene therapy and / or surgery to treat hyperproliferative diseases, including cancer. Such combination therapy involves mixing one or more small molecule IgE inhibitors with one or more additional ingredients known to be effective in reducing one or more symptoms of a disease state. In one variation, the benzimidazole compounds disclosed herein may be administered separately from the additional drug during the same course of the disease state, wherein both the benzimidazole compound and the emollient compound are administered according to their respective effective treatment methods. Can be.

While many preferred embodiments of the invention and variations thereof have been described in detail, other variations and methods of use will be apparent to those skilled in the art. Accordingly, it should be understood that various adaptations, modifications and substitutions may be made to the equivalents without departing from the spirit or scope of the invention.

Claims (34)

A pharmaceutical composition for treating or preventing allergic reactions associated with increased IgE levels in mammals, or inhibiting cell proliferation, comprising one or more of the following compounds: ; ; And ; In the above, R is selected from the group consisting of H, C ₁ -C ₅ alkyl, benzyl, p-fluorobenzyl, and dialkylamino alkyl, and C ₁ -C ₅ alkyl is from the group consisting of linear, branched, or cyclic alkyl Selected; R ₁ and R ₂ are independently H, alkyl, substituted alkyl, C ₃ -C ₉ cycloalkyl, substituted C ₃ -C ₉ cycloalkyl, polycyclic aliphatic groups, phenyl, substituted phenyl, naphthyl, substituted Selected from the group consisting of naphthyl, heteroaryl, and substituted heteroaryl, wherein the heteroaryl and substituted heteroaryl contain 1-3 heteroatoms, wherein the heteroatoms are independently composed of nitrogen, oxygen, and sulfur Selected from the group; Substituted phenyl, substituted naphthyl, and substituted heteroaryl contain 1-3 substituents, wherein the substituents are H, halogen, polyhalogen, alkoxy group, substituted alkoxy, alkyl, substituted alkyl, dialkylamino Alkyl, hydroxyalkyl, OH, OCH ₃ , COOH, COOR ', COR', CN, CF ₃ , OCF ₃ , NO ₂ , NR'R ', NHCOR', and CONR'R '; R ₃ and R ₄ are independently selected from the group consisting of H, alkyl, aryl, heteroaryl, and COR ′; R 'is H, alkyl, substituted alkyl, C ₃ -C ₉ cycloalkyl, substituted C ₃ -C ₉ cycloalkyl, polycyclic aliphatic group, phenyl, substituted phenyl, naphthyl, substituted naphthyl, heteroaryl , And substituted heteroaryl, wherein the heteroaryl and substituted heteroaryl contain 1-3 heteroatoms, and the heteroatoms are independently selected from the group consisting of nitrogen, oxygen, and sulfur; X and Y are independently H, halogen, alkoxy, substituted alkoxy, alkyl, substituted alkyl, dialkylaminoalkyl, hydroxyalkyl, OH, OCH ₃ , COOH, CN, CF ₃ , OCF ₃ , NO ₂ , COOR '', CHO, and COR ''; R '' is C ₁ -C ₈ alkyl, wherein C ₁ -C ₈ alkyl is selected from the group consisting of linear, branched, or cyclic alkyl. A compound according to claim 1, wherein the polycyclic aliphatic group is selected from the group consisting of adamantyl, bicycloheptyl, camphoryl, bicyclo [2,2,2] octanyl and norbornyl. The compound of claim 1, wherein the heteroaryl and substituted heteroaryl are pyridine, thiazole, isothiazole, oxazole, pyrimidine, pyrazine, furan, thiophene, isoxazole, pyrrole, pyridazine, 1,2,3- Triazine, 1,2,4-triazine, 1,3,5-triazine, pyrazole, imidazole, indole, quinoline, isoquinoline, benzothiopine, benzofuran, parathiazine, pyran, chromen, Compound selected from the group consisting of pyrrolidine, pyrazolidine, imidazolidine, morpholine, thiomorpholine, and corresponding heterocyclic groups. The pharmaceutical composition of claim 1, further comprising one or more additional ingredients having the activity of reducing one or more symptoms associated with allergic reactions or cell proliferation. Use for the manufacture of a medicament for the treatment or prevention of an allergic reaction in a mammal caused by an increase in IgE levels of a compound according to claim 1. 6. Use according to claim 5, further comprising administering one or more additional ingredients having the activity of reducing one or more symptoms associated with the allergic reaction. 7. The method of claim 6, wherein the one or more additional ingredients are selected from an immediate β ₂ -adrenergic agonist, a sustained β ₂ -adrenergic agonist, an antihistamine, a phosphodiesterase inhibitor, an anticholinergic agent, a corticosteroid, an inflammatory mediator release inhibitor, a leukotriene receptor antagonist. Use, characterized in that selected from the group consisting of. Use according to claim 6, characterized in that one or more additional ingredients are combined with one or more IgE-inhibiting compounds in a pharmaceutically acceptable diluent and administered together to the mammal. Use according to claim 8, characterized in that at least one IgE inhibiting compound is administered at a dose of about 0.01 to about 100 mg / kg body weight per day. Use according to claim 9, characterized in that the dose is administered in divided doses at regular intervals. Use according to claim 10, characterized in that a constant cycle occurs every day. Use for the manufacture of a medicament for the treatment or prevention of asthma in a mammal wherein the allergic reaction of the compound according to claim 1 is caused by an increase in IgE levels. 13. Use according to claim 12, further comprising administering one or more additional ingredients having the activity of reducing one or more symptoms associated with asthma. 14. The group of claim 13, wherein the additional component consists of an immediate beta ₂ -adrenergic agonist, a sustained beta ₂ -adrenergic agonist, an antihistamine, a phosphodiesterase inhibitor, an anticholinergic agent, a corticosteroid, an inflammatory mediator release inhibitor, a leukotriene receptor antagonist Use, characterized in that selected from. Use of a compound according to claim 1 for the manufacture of a medicament for inhibiting cell proliferation in a mammal. 16. The use of claim 15, further comprising administering one or more additional ingredients having the activity of reducing one or more symptoms associated with cell proliferation. 17. The use of claim 16, wherein the one or more additional ingredients are selected from the group consisting of antifungal, antiviral, antibiotic, anti-inflammatory, anticancer agents. 17. The method of claim 16, wherein the one or more additional ingredients are selected from the group consisting of alkylating agents, metabolites, DNA cutters, topoisomerase I activity inhibitors, topoisomerase II activity inhibitors, DNA binders, and spindle inhibitors. Use. Use according to claim 16, characterized in that one or more additional ingredients are combined with one or more compounds according to claim 1 in a pharmaceutically acceptable diluent and administered together to the mammal. 20. The use of claim 19, wherein the one or more compounds of claim 1 are administered at a dose of about 0.01 to 100 mg / kg body weight per day. The use of claim 20, wherein the dose is administered in divided doses at regular intervals. Use according to claim 21, characterized in that a constant cycle takes place daily. The pharmaceutical composition of claim 1, wherein the compound is selected from the group: The pharmaceutical composition of claim 1, wherein the compound is selected from the group: The pharmaceutical composition of claim 1, wherein the compound is selected from the group: The pharmaceutical composition of claim 1, wherein the compound is selected from the group: The pharmaceutical composition of claim 1, wherein the compound is selected from the group: The pharmaceutical composition of claim 1, wherein the compound is selected from the group: The pharmaceutical composition of claim 1, wherein the compound is selected from the group: The pharmaceutical composition of claim 1, wherein the compound is selected from the group: The pharmaceutical composition of claim 1, wherein the compound is selected from the group: Reacting 3,4-diaminobenzoic acid with 4-nitrobenzaldehyde to obtain a first intermediate or salt thereof, amination of the first intermediate or salt thereof to obtain a first intermediate or salt thereof, and a second Compounds having the formulas below, wherein the intermediates or salts thereof are reduced to yield a third intermediate or salts thereof, and acylated third intermediates or salts thereof to obtain a compound having the formula or a salt thereof How to prepare a salt of: In the above, R is selected from the group consisting of H, C ₁ -C ₅ alkyl, benzyl, p-fluorobenzyl, and dialkylamino alkyl, and C ₁ -C ₅ alkyl is from the group consisting of linear, branched, or cyclic alkyl Selected; R ₁ and R ₂ are independently H, alkyl, substituted alkyl, C ₃ -C ₉ cycloalkyl, substituted C ₃ -C ₉ cycloalkyl, polycyclic aliphatic groups, phenyl, substituted phenyl, naphthyl, substituted Selected from the group consisting of naphthyl, heteroaryl, and substituted heteroaryl, wherein the heteroaryl and substituted heteroaryl contain 1-3 heteroatoms, wherein the heteroatoms are independently composed of nitrogen, oxygen, and sulfur Selected from the group; Substituted phenyl, substituted naphthyl, and substituted heteroaryl contain 1-3 substituents, wherein the substituents are H, halogen, polyhalogen, alkoxy group, substituted alkoxy, alkyl, substituted alkyl, dialkylamino Alkyl, hydroxyalkyl, OH, OCH ₃ , COOH, COOR ', COR', CN, CF ₃ , OCF ₃ , NO ₂ , NR'R ', NHCOR', and CONR'R '; R ₃ and R ₄ are independently selected from the group consisting of H, alkyl, aryl, heteroaryl, and COR ′; R 'is H, alkyl, substituted alkyl, C ₃ -C ₉ cycloalkyl, substituted C ₃ -C ₉ cycloalkyl, polycyclic aliphatic groups, phenyl, substituted phenyl, naphthyl, substituted naphthyl, heteroaryl , And substituted heteroaryl, wherein the heteroaryl and substituted heteroaryl contain 1-3 heteroatoms, and the heteroatoms are independently selected from the group consisting of nitrogen, oxygen, and sulfur; X and Y are independently H, halogen, alkoxy, substituted alkoxy, alkyl, substituted alkyl, dialkylaminoalkyl, hydroxyalkyl, OH, OCH ₃ , COOH, CN, CF ₃ , OCF ₃ , NO ₂ , COOR '', CHO, and COR ''; R '' is C ₁ -C ₈ alkyl, wherein C ₁ -C ₈ alkyl is selected from the group consisting of linear, branched, or cyclic alkyl. Reacting 4-nitro-1,2-phenylenediamine with alkyl 4-formylbenzoate to obtain a first intermediate or salt thereof, and treating the first intermediate or salt thereof with an aqueous base to form a second intermediate or Obtaining salts thereof, amination of a second intermediate or salt thereof to obtain a third intermediate or salt thereof, reduction of the third intermediate or salt thereof to obtain a fourth intermediate or salt thereof, A process for preparing a compound having the formula or a salt thereof, wherein the fourth intermediate or salt thereof is acylated to yield a compound having the formula or a salt thereof: In the above, R is selected from the group consisting of H, C ₁ -C ₅ alkyl, benzyl, p-fluorobenzyl, and dialkylamino alkyl, and C ₁ -C ₅ alkyl is from the group consisting of linear, branched, or cyclic alkyl Selected; R ₁ and R ₂ are independently H, alkyl, substituted alkyl, C ₃ -C ₉ cycloalkyl, substituted C ₃ -C ₉ cycloalkyl, polycyclic aliphatic groups, phenyl, substituted phenyl, naphthyl, substituted Selected from the group consisting of naphthyl, heteroaryl, and substituted heteroaryl, wherein the heteroaryl and substituted heteroaryl contain 1-3 heteroatoms, wherein the heteroatoms are independently composed of nitrogen, oxygen, and sulfur Selected from the group; Substituted phenyl, substituted naphthyl, and substituted heteroaryl contain 1-3 substituents, wherein the substituents are H, halogen, polyhalogen, alkoxy group, substituted alkoxy, alkyl, substituted alkyl, dialkylamino Alkyl, hydroxyalkyl, OH, OCH ₃ , COOH, COOR ', COR', CN, CF ₃ , OCF ₃ , NO ₂ , NR'R ', NHCOR', and CONR'R '; R ₃ and R ₄ are independently selected from the group consisting of H, alkyl, aryl, heteroaryl, and COR ′; R 'is H, alkyl, substituted alkyl, C ₃ -C ₉ cycloalkyl, substituted C ₃ -C ₉ cycloalkyl, polycyclic aliphatic groups, phenyl, substituted phenyl, naphthyl, substituted naphthyl, heteroaryl , And substituted heteroaryl, wherein the heteroaryl and substituted heteroaryl contain 1-3 heteroatoms, and the heteroatoms are independently selected from the group consisting of nitrogen, oxygen, and sulfur; X and Y are independently H, halogen, alkoxy, substituted alkoxy, alkyl, substituted alkyl, dialkylaminoalkyl, hydroxyalkyl, OH, OCH ₃ , COOH, CN, CF ₃ , OCF ₃ , NO ₂ , COOR '', CHO, and COR ''; R '' is C ₁ -C ₈ alkyl, wherein C ₁ -C ₈ alkyl is selected from the group consisting of linear, branched, or cyclic alkyl. Reacting 3,4-diaminobenzoic acid with 4-alkoxycarbonyl benzaldehyde to obtain a first intermediate or salt thereof, wherein the first intermediate or salt thereof is selected from the group of inorganic acid halides, organic acid chlorides, and mixed anhydrides To obtain a second intermediate or salt thereof, amination of the second intermediate or salt thereof to obtain a third intermediate or salt thereof, and treatment of the third intermediate or salt thereof with an aqueous base to 4 A process for preparing compounds having the following formula or salts thereof, wherein the intermediate or salts thereof are obtained and the fourth intermediate or salts thereof are aminated to obtain a compound having the formula or a salt thereof: In the above, R is selected from the group consisting of H, C ₁ -C ₅ alkyl, benzyl, p-fluorobenzyl, and dialkylamino alkyl, and C ₁ -C ₅ alkyl is from the group consisting of linear, branched, or cyclic alkyl Selected; R ₁ and R ₂ are independently H, alkyl, substituted alkyl, C ₃ -C ₉ cycloalkyl, substituted C ₃ -C ₉ cycloalkyl, polycyclic aliphatic groups, phenyl, substituted phenyl, naphthyl, substituted Selected from the group consisting of naphthyl, heteroaryl, and substituted heteroaryl, wherein the heteroaryl and substituted heteroaryl contain 1-3 heteroatoms, wherein the heteroatoms are independently composed of nitrogen, oxygen, and sulfur Selected from the group; Substituted phenyl, substituted naphthyl, and substituted heteroaryl contain 1-3 substituents, wherein the substituents are H, halogen, polyhalogen, alkoxy group, substituted alkoxy, alkyl, substituted alkyl, dialkylamino Alkyl, hydroxyalkyl, OH, OCH ₃ , COOH, COOR ', COR', CN, CF ₃ , OCF ₃ , NO ₂ , NR'R ', NHCOR', and CONR'R '; R ₃ and R ₄ are independently selected from the group consisting of H, alkyl, aryl, heteroaryl, and COR ′; R 'is H, alkyl, substituted alkyl, C ₃ -C ₉ cycloalkyl, substituted C ₃ -C ₉ cycloalkyl, polycyclic aliphatic groups, phenyl, substituted phenyl, naphthyl, substituted naphthyl, heteroaryl , And substituted heteroaryl, wherein the heteroaryl and substituted heteroaryl contain 1-3 heteroatoms, and the heteroatoms are independently selected from the group consisting of nitrogen, oxygen, and sulfur; X and Y are independently H, halogen, alkoxy, substituted alkoxy, alkyl, substituted alkyl, dialkylaminoalkyl, hydroxyalkyl, OH, OCH ₃ , COOH, CN, CF ₃ , OCF ₃ , NO ₂ , COOR '', CHO, and COR ''; R '' is C ₁ -C ₈ alkyl, wherein C ₁ -C ₈ alkyl is selected from the group consisting of linear, branched, or cyclic alkyl.