HK1130249A - Diphenylurea derivatives and their use as chloride channel blockers or bkca channel modulators - Google Patents

Description

Diphenylurea derivatives and their use as chloride channel blockers or BKCaApplication of channel regulator

Technical Field

The present invention relates to novel diphenylurea derivatives useful as chloride channel blockers or BKCa channel modulators.

In other aspects the invention relates to the use of these compounds in therapy and to pharmaceutical compositions comprising the compounds of the invention.

Background

Chloride channels can be adapted for a variety of specific cellular functions and promote normal functions such as skeletal muscle and smooth muscle cells. Chloride channels may be found in every cell, from bacteria to mammals. Its physiological tasks range from regulation of cell volume to stabilization of membrane potential, epithelial or transcellular transport and acidification of intracellular organelles.

Likewise, Ca²⁺Activated BK channels are present in a number of cells, including most central and peripheral nerve cells, striated muscle cells, cardiac muscle cells, airway smooth muscle, vasculature, gastrointestinal tractTract and bladder cells, including pancreatic b-cells and cells in the renal tubules, are included in the endocrine and exocrine glands.

For providing blockers or BK with chloride channels_CaThere is a continuing need for compounds that modulate the activity of channels and have optimal pharmacological properties. Furthermore, there is a strong need to find effective compounds without the undesirable side effects associated with earlier compounds.

The use of ERG channel openers in the treatment of hyperexcitability-related neuronal disorders and arrhythmias is described in WO 2005/023237 and WO 2005/023238(Poseidon pharmaceuticals A/S). Among the compounds disclosed are 1- (3-trifluoromethyl-phenyl) -3- [2- (5-oxo-4, 5-dihydro- [1, 2, 4] oxadiazol-3-yl) -4- (4' -N, N-dimethyl-carbamoyl) -biphenyl ] -urea.

Summary of The Invention

It is an object of the present invention to provide compounds which are chloride channel blockers or BK_CaNovel compounds which act as channel modulators.

It is another object of the present invention to provide compounds with better selectivity. It is another object of the present invention to provide compounds with better efficacy.

Another object of the present invention is to provide a chloride channel or BK specific to a cell or tissue_CaA compound in which a channel functions. It is another object of the invention to provide compounds that act on specific families or subtypes of chloride channels.

It is another object of the present invention to provide compounds with more optimized pharmacodynamic properties, such as kinetic behavior, bioavailability, solubility and efficacy.

The invention provides in a first aspect thereof a compound of formula I,

any of its isomers or any mixture of its isomers or a pharmaceutically acceptable salt thereof, wherein R is^o，R^m，R^p，R²，R³，R⁴，R⁵And R⁶As defined below.

In its second aspect, the present invention provides a pharmaceutical composition comprising a therapeutically effective amount of a compound of the present invention, any of its isomers or any mixture of its isomers, or a pharmaceutically acceptable salt thereof, together with at least one pharmaceutically acceptable carrier, excipient or diluent.

In another aspect, the invention provides the use of a compound of the invention, any of its isomers or any mixture of its isomers, or a pharmaceutically acceptable salt thereof, for the preparation of a pharmaceutical composition for the treatment, prevention or alleviation of a disease or a disorder or a condition of a mammal, including a human, which disease, disorder or condition is the blockade of a chloride channel or the modulation of BK_CaThe channel has a response; or in the preparation of a medicament for increasing blood brain barrier permeability.

In another aspect the invention relates to a method of treatment, prevention or alleviation of a disease or a disorder or a condition of a living animal body, including a human, which disorder, disease or condition is the blockade of a chloride channel or the modulation of BK_CaThe channel is responsive and the method comprises the step of administering to such a living animal body in need thereof a therapeutically effective amount of a compound of the present invention, any of its isomers or any mixture of its isomers, or a pharmaceutically acceptable salt thereof.

Other objects of the present invention will be apparent to those skilled in the art from the following detailed description and examples.

Detailed Description

Diphenyl urea derivatives

The present invention provides in a first aspect thereof a chemical compound of formula I,

any of its isomers or any mixture of its isomers, or a pharmaceutically acceptable salt thereof; wherein

R^o，R^m，R^p，R²，R³，R⁴，R⁵And R⁶Independently of each other represent

Hydrogen, halogen, trifluoromethyl, trifluoromethoxy, alkyl or alkoxy.

In one embodiment of the compounds of formula I, R^oRepresents hydrogen; r^mRepresents hydrogen; and R is^pRepresents halogen, trifluoromethyl, trifluoromethoxy, alkyl or alkoxy. In another embodiment, R^oRepresents hydrogen; r^mRepresents hydrogen; and R is^pRepresents halogen or trifluoromethyl. In a particular embodiment, R^pRepresents a trifluoromethyl group.

In another embodiment, R³，R⁴，R⁵And R⁶Represents hydrogen; and R is²Represents halogen, trifluoromethyl, trifluoromethoxy, alkyl or alkoxy. In a particular embodiment, R²Represents halogen such as chlorine.

In another embodiment, R²，R⁴，R⁵And R⁶Represents hydrogen; and R is³Represents halogen, trifluoromethyl, trifluoromethoxy, alkyl or alkoxy. In a particular embodiment, R³Represents halogen, such as bromine.

In another embodiment, R²，R⁵And R⁶Represents hydrogen; and R is³And R⁴Independently of one another, halogen, trifluoromethyl, trifluoromethoxy, alkyl or alkoxyAnd (4) a base. In another embodiment, R³And R⁴Independently of one another, represents halogen or trifluoromethyl. In a particular embodiment, R³And R⁴One represents halogen, such as chlorine, and R³And R⁴The other of (a) represents a trifluoromethyl group. In another embodiment, R³Represents trifluoromethyl and R⁴Represents halogen such as chlorine.

In another embodiment, R²，R⁴And R⁶Represents hydrogen; and R is³And R⁵Independently of one another, represents halogen, trifluoromethyl, trifluoromethoxy, alkyl or alkoxy. In a particular embodiment, R³And R⁵Independently of one another, represent halogen. In another embodiment, R³Represents fluorine and R⁵Represents fluorine.

In a particular embodiment, the compounds of the invention are:

n- (3, 5-difluoro-phenyl) -N '- [3- (5-oxo-4, 5-dihydro- [1, 2, 4] oxadiazol-3-yl) -4' -trifluoromethyl-biphenyl-4-yl ] urea;

n- (4-chloro-3-trifluoromethyl-phenyl) -N '- {3- (5-oxo-4, 5-dihydro- [1, 2, 4] -oxadiazol-3-yl) -4' -trifluoromethyl-biphenyl-4-yl } urea;

n- (3-bromo-phenyl) -N '- {3- (5-oxo-4, 5-dihydro- [1, 2, 4] -oxadiazol-3-yl) -4' -trifluoromethyl-biphenyl-4-yl } urea;

n- (2-chloro-phenyl) -N '- {3- (5-oxo-4, 5-dihydro- [1, 2, 4] -oxadiazol-3-yl) -4' -trifluoromethyl-biphenyl-4-yl } urea;

or a pharmaceutically acceptable salt thereof.

Definition of substituents

In the context of the present invention, halogen denotes fluorine, chlorine, bromine or iodine.

Alkyl means straight or branched chain of 1 to 6 carbon atoms including, but not limited to, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, pentyl and hexyl; methyl, ethyl, propyl and isopropyl are preferred groups.

Alkoxy is O-alkyl, wherein alkyl is as defined above.

Pharmaceutically acceptable salts

The chemical compounds of the present invention may be provided in any form suitable for the intended administration. Suitable forms include pharmaceutically (i.e. physiologically) acceptable salts of the chemical compounds of the invention as well as prodrug (predug) or prodrug (produg) forms.

Examples of pharmaceutically acceptable addition salts include, but are not limited to, non-toxic inorganic and organic acid addition salts such as hydrochloride, hydrobromide, nitrate, perchlorate, phosphate, sulfate, formate, acetate, aconate, ascorbate, benzenesulfonate, benzoate, cinnamate, citrate, pamoate, heptanoate, fumarate, glutamate, glycolate, lactate, maleate, malonate, mandelate, methanesulfonate, naphthalene-2-sulfonate, phthalate, salicylate, sorbate, stearate, succinate, tartrate, p-toluenesulfonate and the like. Such salts may be formed by procedures well known and described in the art.

Other acids, such as oxalic acid, which are not considered pharmaceutically acceptable, may be used to prepare salts useful as intermediates in obtaining the chemical compounds of the invention and their pharmaceutically acceptable acid addition salts.

Examples of pharmaceutically acceptable cationic salts of the compounds of the present invention include, but are not limited to, sodium, potassium, calcium, magnesium, zinc, aluminum, lithium, choline, lysinium (lysinium), ammonium salts and the like of the compounds of the present invention containing an anionic group. Such cationic salts can be formed by procedures well known and described in the art.

In the context of the present invention, an "onium salt" of an N-containing compound is also considered to be a pharmaceutically acceptable salt. Preferred "onium salts" include alkyl-onium salts, cycloalkyl-onium salts, and cycloalkylalkyl-onium salts.

Examples of prodrugs or prodrug forms of chemical compounds of the invention include examples of suitable prodrugs of the substances of the invention, including compounds which are modified at one or more reactive or derivatizable groups of the parent compound. Of particular interest are compounds modified at the carboxyl, hydroxyl or amino group. Examples of suitable derivatives are esters or amides.

The chemical compounds of the present invention may be prepared in soluble or insoluble form with pharmaceutically acceptable solvents such as water, ethanol, and the like. Soluble forms may also include hydrated forms such as the monohydrate, dihydrate, hemihydrate, trihydrate, tetrahydrate and the like. In general for the purposes of the present invention, a soluble form is considered to be equivalent to an insoluble form.

Isomers

One skilled in the art will appreciate that the compounds of the present invention may exist in different stereoisomeric forms-including enantiomers, diastereomers and cis-trans isomers.

The present invention includes all such isomers and any mixtures thereof, including racemic mixtures.

Methods for resolving optical isomers well known to those skilled in the art can be used and will be apparent to those of ordinary skill in the art. Such methods include those discussed by j.jaques, a.collet and s.wilen in "enertiomers, racemes and solutions", John Wiley and Sons, New York (1981).

Optically active compounds can also be prepared from optically active starting materials.

Labelled compounds

The compounds of the invention may be used in labelled or unlabelled form. In the context of the present invention, a labeled compound has one or more atoms replaced with an atom having an atomic mass or mass number different from the atomic mass or mass number usually found in nature. The label is conveniently capable of quantitatively detecting the compound.

The labeled compounds of the invention may be used as diagnostic tools, radiotracers or monitoring agents in various diagnostic methods, and for in vivo receptor imaging.

The labelled isomer of the invention preferably comprises at least one radionuclide as label. Positron emitting radionuclides are all candidates for use. In the context of the present invention, the radionuclide is preferably selected from²H (deuterium),³H (tritium),¹³C、¹⁴C、¹³¹I、¹²⁵I、¹²³I and¹⁸F。

the physical method for detecting the labelled isomer of the invention may be selected from Positron Emission Tomography (PET), Single Photon Emission Computed Tomography (SPECT), Magnetic Resonance Spectroscopy (MRS), Magnetic Resonance Imaging (MRI) and computer assisted axial tomography (CAT) or a combination thereof.

Preparation method

The compounds of the present invention may be prepared by conventional methods for chemical synthesis, such as those described in the preparation examples. The starting materials for the processes described herein are well known or readily prepared by conventional methods from commercially available chemicals.

Conventional methods may also be used to convert one compound of the invention to another compound of the invention.

The reaction end product described herein may be isolated by conventional techniques, e.g., by extraction, crystallization, distillation, chromatography, and the like.

Biological activity

The compounds of the invention are useful as chloride channel blockersOr BK_CaA channel modulator.

Preferred compounds of the invention exhibit biological activity in the submicromolar and micromolar range, i.e., less than 1 to about 100. mu.M.

Examples of types of chloride channels are Volume Regulated Anion Channels (VRAC) or chloride channels of osteoclasts or erythrocytes. To determine the activity of these compounds, various chloride channel blocking assays known in the art can be used.

Thus, in another aspect, the compounds of the invention are considered for use in the treatment, prevention or alleviation of a disease or a disorder or a condition which is responsive to blockade of chloride channels.

In a particular embodiment, the compounds of the invention are considered for the treatment, prevention or alleviation of:

bone metabolic diseases such as osteoclast-associated bone diseases such as osteoporosis, postmenopausal osteoporosis, secondary osteoporosis, osteolytic breast cancer bone metastasis, osteolytic cancer invasion (invation) or paget's disease of bone;

diseases responsive to inhibition of angiogenesis, such as diseases involving tumor cell proliferation, such as cancer, metastatic cancer, prostate cancer, lung cancer, breast cancer, bladder cancer, kidney cancer, colon cancer, stomach cancer, pancreatic cancer, ovarian cancer, melanoma, hepatoma, sarcoma, lymphoma;

ocular angiogenesis-related diseases such as exudative macular degeneration, age-related macular degeneration (AMD), retinopathy, diabetic retinopathy, proliferative diabetic retinopathy, Diabetic Macular Edema (DME), ischemic retinopathy (e.g. retinal vein (vain) or artery occlusion), retinopathy of prematurity, neovascular glaucoma, corneal neovascularization; and

diseases, disorders or conditions responsive to decreased intraocular pressure, such as ocular hypertension, open angle glaucoma, chronic open angle glaucoma, angle-closure glaucoma, ciliary congestion due to angle-closure glaucoma; and

rheumatoid arthritis, psoriasis

Sickle cell anemia.

In another aspect, the compounds of the invention are contemplated for use in the treatment, prevention or amelioration of the modulation of BK_CaA disease, disorder or condition in which the channel is responsive.

In a particular embodiment, the compounds of the invention are contemplated for use in the treatment, prevention or alleviation of:

cardiovascular diseases such as atherosclerosis, ischemia, reperfusion injury, hypertension, restenosis, arterial inflammation, myocardial ischemia or ischemic heart disease;

obstructive or inflammatory airway diseases, such as airway hyperresponsiveness, pneumoconiosis, aluminosis, anthracosis, asbestosis, chalicosis, camelina, siderosis, silicosis, tabacosis, gossyposis, sarcoidosis, berylliosis, emphysema, Acute Respiratory Distress Syndrome (ARDS), Acute Lung Injury (ALI), acute or chronic infectious lung disease, Chronic Obstructive Pulmonary Disease (COPD), bronchitis, chronic bronchitis, wheezing bronchitis, exacerbation of airway hyperresponsiveness (excerbation) or cystic fibrosis, cough, including chronic cough, exacerbation of airway hyperresponsiveness, pulmonary fibrosis, pulmonary hypertension, inflammatory lung disease or acute or chronic respiratory infectious disease;

urinary incontinence, psychosis, epilepsy or pain.

Furthermore, the compounds of the present invention may also be suitable for facilitating transport of therapeutic substances across the blood-brain barrier, and in particular for facilitating delivery of chemotherapeutic agents and viral particles to tumor cells and other abnormal brain tissue.

Thus, in another aspect, the invention relates to the use of a compound of the invention as a promoter for increasing blood brain barrier permeability and thereby enabling the promotion of transport of a therapeutic substance across the blood brain barrier, including the blood-tumor barrier found in brain tumors.

In one embodiment of this aspect, the compounds of the invention are useful for promoting the access of an active agent to an area of abnormal brain tissue that is physiologically damaged, traumatized, infected, stroke or ischemia-affected. Such abnormal brain regions are areas of benign or malignant neoplastic tissue or other neoplastic disease or disorder. The malignant tumor may in particular be a glioma, glioblastoma, oligodendroglioma, astrocytoma, ependymoma, primary neuroectodermal tumor, atypical meningioma, malignant meningioma, neuroblastoma, sarcoma, melanoma, lymphoma or carcinoma.

When used as a promoting agent, the compounds of the present invention may be co-administered with the therapeutic agent by any suitable route, in any convenient manner. Preferably, the therapeutic agent is administered simultaneously (i.e., simultaneously or concurrently) or substantially simultaneously (i.e., within about 1 hour, preferably within 30 minutes, and even more preferably within 15 minutes).

The active agents for use in the present invention, i.e., the facilitator and the therapeutic agent, may be administered by any suitable route by which the active agents are delivered to the bloodstream. Preferably, the procedure is performed by intravenous, intramuscular or intraarterial injection or infusion.

The therapeutic agent for use in the present invention may be any active agent or drug. However, preferred therapeutic agents or drugs for use in the present invention are antineoplastic agents, chemotherapeutic agents, cytotoxic agents, DNA expression vectors, proteins, oligonucleotides, nucleotide analogs, antimicrobial agents, interferons, cytokines, cytokine agonists, cytokine antagonists, immunotoxins, immunosuppressive agents, boron compounds, monoclonal antibodies, adrenergic agents, anticonvulsants, antihypertensives, atraumatic agents, anti-traumatic agents, anticancer chemotherapeutic agents and diagnostic agents.

Preferred chemotherapeutic agents for use in the present invention include:

alkylating agents, such as nitrogen mustards (e.g. nitrogen mustards, cyclophosphamide, ifosfamide (ifosamide), melphalan and chlorambucil), vinylideneamines and methylmelamines (e.g. altretamine and thiotepa), alkylsulfonates (e.g. busulfan), nitrosoureas (e.g. carmustine (BCNU), lomustine (CCNU), semustine (methyl-CCNU) and streptozocin), triazenes (e.g. Dacarbazine (DTIC));

antimetabolites, such as folic acid analogs (e.g., methotrexate), pyrimidine analogs (e.g., fluorouracil, floxuridine, and cytarabine), purine analogs, and related inhibitors (e.g., mercaptopurine, thioguanine, and pentostatin); and

natural antimitotic products, such as vinca alkaloids (e.g. vinblastine and vincristine), epipodophyllotoxins (e.g. etoposide and teniposide), antibiotics (e.g. dactinomycin, daunorubicin, doxorubicin, bleomycin, plicamycin and mitomycin), enzymes (e.g. L-asparaginase), platinum coordination complexes (e.g. cisplatin and carboplatin) and biological response modifiers, such as interferons (e.g. interferon- α).

In another preferred embodiment, the DNA expression vector is a viral vector, preferably an adenovirus-derived vector or a herpes simplex virus-derived vector.

In another preferred embodiment, the diagnostic agent for the use according to the invention may in particular be an imaging agent or a contrast agent, and it may in particular be a radiolabeled substance, a gallium-labeled substance or a contrast agent selected from the group consisting of ferrimagnetic agents, fluorescent agents, luminescent agents and iodinated contrast agents.

When used as a facilitator, the compounds of the invention may preferably be co-administered with a therapeutic agent for targeting areas of brain tissue or brain tumour growth areas which are physiologically directly affected by physical or biochemical damage, for example alzheimer's disease, parkinson's syndrome, trauma, infection, stroke, cerebral ischaemia, such as benign or malignant brain tumour tissue.

Method of treatment

Another aspect of the invention provides a method of treatment, prevention or alleviation of a disease or a disorder or a condition of a living animal body, including a human, which disorder, disease or condition is the blockade of a chloride channel or the modulation of BK_CaThe channel is responsive and the method comprises administering to such a living animal body, including a human, in need thereof an effective amount of a compound of the invention.

Preferred medical indications of interest for the present invention are those described above.

It is presently contemplated that suitable dosage ranges are from 0.1 to 1000mg daily, from 10 to 500mg daily, and particularly from 30-100mg daily, depending generally on the exact mode of administration, form of administration, indication for which administration is being made, subject involved and subject weight involved, and further, the preference and experience of the attending physician or veterinarian. When used in combination with compounds known in the art to treat such diseases, dosage regimens may be reduced.

Combination therapy

The use of the compounds of the present invention may be combined with the use of other compounds for the treatment, prevention or alleviation of diseases, disorders or conditions responsive to blockade of chloride channels.

As an example, the compounds may be used in combination with one or more other agents useful in the treatment, prevention, or amelioration of diseases responsive to inhibition of angiogenesis, such as compounds useful in anti-metastatic therapy. Such other drugs include cytotoxic compounds, antimitotic compounds and antimetabolites.

Examples of cytotoxic compounds, including cytotoxic alkylating agents, include carmustine (BCNU), fotemustine, temozolomide (temodal), ifosfamide and cyclophosphamide (cyclophosfamide).

Examples of antimitotic compounds include paclitaxel (taxol) and docetaxel.

Examples of antimetabolites include methotrexate.

In addition, the pharmaceutical compositions used in the present invention may be used or administered in combination with other treatments or therapies. Examples of other treatments or therapies include radiation therapy and surgery.

Furthermore, the use of the compounds of the present invention may be combined with the use of other bone metabolism control compounds for the treatment of bone metabolism diseases. Such known bone metabolism control compounds include bisphosphonates, such as etidronic acid, pamidronic acid or clodronic acid optionally in combination with calcium; estrogen receptor active compounds such as estradiol (oestrogen), i.e. estradiol (oesteradiol) and ethyloesteradiol, calcitonin, 1, 25-dihydroxyvitamin D and its metabolites, fluoride, growth hormone, parathyroid hormone, triiodo-tyrosine (thyrosine), collagen degrading enzymes such as protease inhibitors, or cancer therapeutics.

Furthermore, the use of the compounds of the present invention may be combined with the use of one or more other medicaments for the treatment, prevention or alleviation of a disease or a disorder or a condition that is responsive to the reduction of intraocular pressure. Such other drugs include beta-blockers, parasympathomimetics, miotics, sympathomimetics, and carbonic anhydrase inhibitors.

Furthermore, the use of the compounds of the present invention may be combined with other treatments or therapies.

Treatment of diseases and disorders may be chronic or chronic and treatment of the sudden crisis of the disease and disorder.

Pharmaceutical composition

In another aspect, the present invention provides novel pharmaceutical compositions comprising a therapeutically effective amount of a chemical compound of the present invention.

Although the compounds of the invention for use in therapy may be administered as the starting compounds, it is preferred that the active ingredient, optionally in the form of a physiologically acceptable salt, is incorporated into a pharmaceutical composition together with one or more adjuvants, excipients, carriers, buffers, diluents and/or other conventional pharmaceutical adjuvants.

In a preferred embodiment, the present invention provides a pharmaceutical composition comprising a chemical compound of the present invention or a pharmaceutically acceptable salt or derivative thereof in combination with one or more pharmaceutically acceptable carriers, and optionally in admixture with other therapeutic and/or prophylactic ingredients known and used in the art. The carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not deleterious to the recipient thereof.

The pharmaceutical compositions of the present invention may be those suitable for oral, rectal, bronchial, nasal, pulmonary, topical (including buccal and sublingual), transdermal, vaginal or parenteral (including dermal, subcutaneous, intramuscular, intraperitoneal, intravenous, intraarterial, intracerebral, intraocular injection or infusion) administration or those in a form suitable for administration by inhalation or insufflation, including powder and liquid aerosol administration, or for administration by slow release systems. Examples of suitable sustained release systems include semipermeable matrices of solid hydrophobic polymers containing the compound of the invention, which matrices may be in the form of shaped articles, e.g. films, or microcapsules.

The chemical compounds of the present invention may thus be formulated together with conventional adjuvants, carriers or diluents into pharmaceutical compositions and unit dosage forms thereof. Such forms include solids, and especially tablets, filled capsules, powders and pills, as well as liquids, especially aqueous or non-aqueous solutions, suspensions, emulsions, elixirs and capsules filled with the above, all for oral administration, suppositories for rectal administration, and sterile injectable solutions for parenteral administration. Such pharmaceutical compositions and unit dosage forms thereof may comprise conventional ingredients in conventional proportions, with or without additional active compounds or ingredients, and such unit dosage forms may contain any suitable effective amount of the active ingredient commensurate with the intended daily dosage range of use.

The chemical compounds of the present invention may be administered in a variety of oral and parenteral dosage forms. It will be apparent to those skilled in the art that the dosage forms described below may contain, as the active ingredient, a compound of the present invention or a pharmaceutically acceptable salt of a compound of the present invention.

For preparing pharmaceutical compositions from the compounds of the present invention, the pharmaceutically acceptable carrier may be solid or liquid. Solid form preparations include powders, tablets, pills, capsules, cachets, suppositories, and dispersible granules. A solid carrier can be one or more substances which also act as diluents, flavoring agents, solubilizers, lubricants, suspending agents, binders, preservatives, tablet disintegrating agents, or an encapsulating material.

In powders, the carrier is a finely divided solid which is mixed with the finely divided active component.

In tablets, the active ingredient is mixed with a carrier having the necessary binding capacity in suitable proportions and compacted in the shape and size desired.

Powders and tablets preferably contain from 5% or 10% to about 70% of the active compound. Suitable carriers are magnesium carbonate, magnesium stearate, talc, sugar, lactose, pectin, dextrin, starch, gelatin, tragacanth, methyl cellulose, sodium carboxymethylcellulose, a low melting wax, cocoa butter, and the like. The term "formulation" is intended to include a dosage form of the active compound with encapsulating material as a carrier, the encapsulating material providing a capsule in which the active ingredient, with or without a carrier, is surrounded by a carrier, which is thus in association with the active compound. Similarly, cachets and lozenges are included. Tablets, powders, capsules, pills, cachets, and lozenges can be used as solid forms suitable for oral administration.

To prepare suppositories, a low melting wax, such as a mixture of fatty acid glycerides or cocoa butter, is first melted and the active ingredient is dispersed homogeneously therein by stirring. The molten homogeneous mixture is then poured into a suitably sized mold, allowed to cool and thereby solidify.

Compositions suitable for vaginal administration may be presented as pessaries, tampons, creams, gels, pastes, foams or spray formulations containing in addition to the active ingredient such carriers as are known in the art to be appropriate.

Liquid formulations include solutions, suspensions and emulsions, for example, aqueous or water-propylene glycol solutions. For example, parenteral injection liquid preparations can be formulated as solutions of aqueous polyethylene glycols.

Thus, the chemical compounds of the present invention may be formulated for parenteral administration (e.g., by injection, such as bolus injection or continuous infusion) and may be presented in unit dose form in ampoules, pre-filled syringes, small volume infusion or in multi-dose containers with an added preservative. The compositions may take such forms as suspensions, solutions or emulsions in oily or aqueous vehicles, and may contain formulatory agents such as suspending, stabilizing and/or dispersing agents. Alternatively, the active ingredient may be in powder form, obtained by sterile isolation of a sterile solid or by lyophilization from solution, for constitution with a suitable vehicle, e.g., sterile, pyrogen-free water, before use.

Aqueous solutions suitable for oral use can be prepared by dissolving the active component in water and adding suitable coloring, flavoring, stabilizing and thickening agents as desired.

Aqueous suspensions suitable for oral use can be prepared by dispersing the finely divided active component in water with viscous material, such as natural or synthetic gums, resins, methylcellulose, sodium carboxymethylcellulose, or other well-known suspending agents.

Also included are solid form preparations which are intended to be converted, shortly before use, to liquid form preparations for oral administration. Such liquid forms include solutions, suspensions and emulsions. Such formulations may contain, in addition to the active ingredient, colorants, flavors, stabilizers, buffers, artificial and natural sweeteners, dispersants, thickeners, solubilizing agents, and the like.

For topical application to the epidermis, the compounds of the invention may be formulated as ointments, creams, or lotions, or as a transdermal patch. For example, ointments and creams may be formulated with an aqueous or oily base with the addition of suitable thickening and/or gelling agents. Lotions may be formulated with an aqueous or oily base and will in general also contain one or more emulsifying agents, stabilizing agents, dispersing agents, suspending agents, thickening agents, or coloring agents.

Compositions suitable for topical administration in the oral cavity include lozenges comprising the active ingredient in a flavored basis, usually sucrose and acacia or tragacanth; pastilles comprising the active ingredient in an inert base such as gelatin and glycerin or sucrose and acacia; and mouthwashes comprising the active ingredient in a suitable liquid carrier.

The solution or suspension can be applied directly to the nasal cavity by conventional means, such as with a dropper, pipette or nebulizer. The composition may be provided in a single dose or in multiple doses.

Administration to the respiratory tract may also be achieved by means of an aerosol formulation in which the active ingredient is provided in a pressurised pack together with a suitable propellant, including a chlorofluorocarbon (CFC) such as dichlorodifluoromethane, trichlorofluoromethane or dichlorotetrafluoroethane, carbon dioxide or other suitable gas. The aerosol may suitably also contain a surfactant such as lecithin. The dosage of the drug may be controlled by equipping a metering valve.

Alternatively, the active ingredient may be provided in the form of a dry powder, for example a powder mix of the compound in a suitable powder base such as lactose, starch derivatives such as hydroxypropylmethyl cellulose and polyvinylpyrrolidone (PVP). Suitably, the powder carrier will form a gel within the nasal cavity. The powder compositions may be presented in unit dosage form, for example in the form of capsules or cartridges, such as capsules or cartridges of gelatin, or in the form of blister packs from which the powder may be administered by means of an inhaler.

In compositions intended for administration to the respiratory tract, including intranasal compositions, the compounds will generally have a small particle size, for example of the order of 5 microns or less. Such particle sizes may be obtained by means of methods known in the art, for example by micronization.

If desired, compositions suitable for providing sustained release of the active ingredient may be employed.

The pharmaceutical preparation is preferably in unit dosage form. In such forms, the preparation is subdivided into unit doses containing appropriate quantities of the active component. The unit dosage form can be a packaged preparation, the package containing discrete quantities of preparation, such as packeted tablets, capsules, and powders in vials or ampoules. In addition, the unit dosage form can be a capsule, tablet, cachet, or lozenge itself, or it can be the packaged form of a suitable number of any of these dosage forms.

Tablets or capsules for oral administration and liquids for intravenous administration as well as continuous infusion solutions are preferred compositions.

More detailed information on the formulation and administration techniques can be found in the latest editionReminaton′s Pharmaceutical Sciences(Maack publishing Co. Easton, Pa.).

A therapeutically effective dose refers to an amount of active ingredient that ameliorates a symptom or condition. Therapeutic efficacy and toxicity, e.g. ED₅₀And LD₅₀Can be determined by standard pharmacological procedures in cell cultures or experimental animals. The dose ratio between therapeutic and toxic effects is the therapeutic index and can be determined by the ratio LD₅₀/ED₅₀And (4) showing. Pharmaceutical compositions exhibiting a large therapeutic index are preferred.

The dose administered must of course be carefully adjusted to the age, weight and condition of the individual being treated, as well as the route of administration, dosage form and regimen, and the desired result, and the exact dose should of course be determined by a physician.

The actual dosage depends on the nature and severity of the condition being treated and, within the judgment of the practitioner, may be varied in response to the dosage in accordance with the particular circumstances of the invention to produce the desired therapeutic effect. However, it is presently contemplated that pharmaceutical compositions containing from about 0.1 to about 500mg, preferably from about 1 to about 100mg, more preferably from about 1 to about 10mg of active ingredient per single dose are suitable for therapeutic treatment.

The active ingredient may be administered in one or several doses per day. In some cases satisfactory results can be obtained at doses as low as 0.1. mu.g/kg (intravenous) and 1. mu.g/kg (oral). The upper limit of the dosage range is currently considered to be about 10mg/kg (intravenous) and 100mg/kg (oral). Preferred ranges are from about 0.1 μ g/kg to about 10 mg/kg/day (intravenous) and from about 1 μ g/kg to about 100 mg/kg/day (oral).

Examples

The invention is further explained with reference to the following examples, which are not intended to limit the scope of the claimed invention in any way.

Example 1

4-amino-4' -trifluoromethyl-biphenyl-3-carbonitrile

26.5g of 4- (trifluoromethyl) phenylboronic acid, 25g of 2-amino-5-bromo-benzonitrile and 57.9g of potassium carbonate are mixed in 250ml of water and 500ml of 1, 2-dimethoxyethane. The mixture was bubbled through nitrogen for 10 minutes, then 1g of bis (triphenylphosphine) palladium (II) chloride was added and the reaction mixture was heated at reflux overnight. The reaction mixture was cooled to room temperature, 700ml of water was added and extracted with 800ml of ethyl acetate. The organic phase is washed first with 300ml of saturated sodium chloride, then with 300ml of 2M calcium chloride and finally with 300ml of water. The organic phase was dried over magnesium sulfate, evaporated to an oil and triturated with ether. Yield 30.5g (92%).

Example 2

4-amino-N-hydroxy-4' -trifluoromethyl-biphenyl-3-carboxamide

4-amino-4' -trifluoromethyl-biphenyl-3-carbonitrile (10g) was dissolved in 250ml of methanol and 5.6g of hydroxylamine hydrochloride and 8.1g of triethylamine were added. The reaction mixture was stirred at 50 ℃ overnight and 100ml water and 100ml ethyl acetate were added. The aqueous phase is extracted with 100ml of ethyl acetate. The organic phase is washed with 100ml of water and thereafter with 100ml of saturated sodium chloride. The organic phase was dried over magnesium sulfate and evaporated to an oil. Yield 10.8g (96%).

Example 3

3- (4-amino-4' -trifluoromethyl-biphenyl-3-yl) -4H- [1, 2, 4] -oxadiazol-5-one

Sodium (1.6g) was stirred in 200ml of dry ethanol until it was dissolved, to this solution 10.8g of 4-amino-N-hydroxy-4' -trifluoromethyl-biphenyl-3-carboxamide and 16g of diethyl carbonate were added, the reaction mixture was stirred at 85 ℃ overnight, evaporated to an oil, the residue was dissolved in ethyl acetate and the aqueous phase was extracted with 4N aqueous NaOH. The aqueous phase was acidified with concentrated hydrochloric acid and extracted with ethyl acetate. The aqueous phase was washed with saturated aqueous sodium chloride solution, dried over magnesium sulfate and evaporated to an oil. The residue was dissolved in 150ml boiling ethanol. The solution was cooled to room temperature and the product crystallized, isolated by filtration and dried. Yield 5.1g (45%).

Example 4

N- (3, 5-difluoro-phenyl) -N '- [3- (5-oxo-4, 5-dihydro- [1, 2, 4] oxadiazol-3-yl) -4' -trifluoromethyl-biphenyl-4-yl ] urea

5g3- (4-amino-4' -trifluoromethyl-biphenyl-3-yl) -4H- [1, 2, 4] oxadiazol-5-one were suspended in 100ml dry toluene and 2.5g 3, 5-difluorophenyl isocyanate was added and the reaction mixture was stirred at room temperature overnight, 1.3g 3, 5-difluorophenyl isocyanate was added and stirring was continued overnight. 3, 5-difluorophenylisocyanate (1.3g) and 100ml acetonitrile were added and the reaction mixture was stirred at room temperature for 90 minutes and evaporated to dryness. The residue was dissolved in 100ml boiling acetone, then cooled to 0 ℃ and filtered. The precipitate was recrystallized from 200ml of ethanol, and after filtration, the solution was added to 400ml of water while heating. The product was isolated by filtration. Yield 4g (54%) mp. >150 ℃ decomposition.

Prepared in a similar manner:

n- (4-chloro-3-trifluoromethyl-phenyl) -N '- {3- (5-oxo-4, 5-dihydro- [1, 2, 4] -oxadiazol-3-yl) -4' -trifluoromethyl-biphenyl-4-yl } urea. Mp >139 ℃ decomposition

N- (3-bromo-phenyl) -N '- {3- (5-oxo-4, 5-dihydro- [1, 2, 4] -oxadiazol-3-yl) -4' -trifluoromethyl-biphenyl-4-yl } urea. Mp >200 ℃ decomposition

N- (2-chloro-phenyl) -N '- {3- (5-oxo-4, 5-dihydro- [1, 2, 4] -oxadiazol-3-yl) -4' -trifluoromethyl-biphenyl-4-yl } urea. Mp220-222 ℃.

Example 5

In vitro human red blood cell chloride conductance

Chloride conductance was determined according to the procedure as described in the WO 00/24707 specification, paragraph "Biology". The parameter calculated from these studies was IC₅₀Value-50% of the concentration at which chloride channels are blocked.

The first compound of example 4, N- (3, 5-difluoro-phenyl) -N' - [3- (5-oxo-4, 5-dihydro- [1, 2, 4]Oxadiazoles as fungicides-3-yl) -4' -trifluoromethyl-biphenyl-4-yl]Urea showed an IC of 0.28. mu.M₅₀The value is obtained.

Example 6

Volume modulated anion channel (VRAC) effect

The activity on volume-regulated anion channels (VRAC) was tested by the whole-cell patch clamp technique using human embryonic kidney cells (HEK293) as described by Helix et al in J Membr biol.2003196 (2): 83-94. Briefly, VRAC was activated by swelling cells in hypotonic (75% tonicity) extracellular saline solution and the anionic current induced by voltage clamps (voltage ramps) was measured as a function of time. After the current is stabilized, adding a test compound to the extracellular solution; and time-dependent blocking occurs, calculating K_DThe value is obtained.

The first compound of example 4, N- (3, 5-difluoro-phenyl) -N' - [3- (5-oxo-4, 5-dihydro- [1, 2, 4]Oxadiazol-3-yl) -4' -trifluoromethyl-biphenyl-4-yl]Urea showed a K of 0.23. mu.M_DThe value is obtained.

Example 7

Human BK channel screening

The effect of compounds on membrane current was measured electrophysiologically on Xenopus Oocytes (Xenopus cells) capable of expressing human BK channels, and the current through the channels was recorded using conventional two-electrode voltage clamping.

This assay was performed on the first compound of example 4, N- (3, 5-difluoro-phenyl) -N '- [3- (5-oxo-4, 5-dihydro- [1, 2, 4] oxadiazol-3-yl) -4' -trifluoromethyl-biphenyl-4-yl ] urea, at a concentration of 0.3 μ M of the test compound, and it resulted in an increase in BK current of more than 200% over the base current.

Claims (13)

1. A compound of formula (I):

any of its isomers or any mixture of its isomers,

or a pharmaceutically acceptable salt thereof;

wherein

R^o，R^m，R^p，R²，R³，R⁴，R⁵And R⁶Independently of one another, represents hydrogen, halogen, trifluoromethyl, trifluoromethoxy, alkyl or alkoxy.

2. The compound of claim 1, wherein

R^oRepresents hydrogen;

R^mrepresents hydrogen; and is

R^pRepresents halogen, trifluoromethyl, trifluoromethoxy, alkyl or alkoxy.

3. The compound of claim 1, wherein

R^oRepresents hydrogen;

R^mrepresents hydrogen; and is

R^pRepresents halogen or trifluoromethyl.

4. A compound according to any one of claims 1 to 3, wherein

R³，R⁴，R⁵And R⁶Represents hydrogen; and is

R²Represents halogen, trifluoromethyl, trifluoromethoxy, alkyl or alkoxy.

5. A compound according to any one of claims 1 to 3, wherein

R²，R⁴，R⁵And R⁶Represents hydrogen; and is

R³Represents halogen, trifluoromethyl, trifluoromethoxy, alkyl or alkoxy.

6. A compound according to any one of claims 1 to 3, wherein

R²，R⁵And R⁶Represents hydrogen; and is

R³And R⁴Independently of one another, represents halogen, trifluoromethyl, trifluoromethoxy, alkyl or alkoxy.

7. A compound according to any one of claims 1 to 3, wherein

R²，R⁴And R⁶Represents hydrogen; and is

R³And R⁵Independently of one another, represents halogen, trifluoromethyl, trifluoromethoxy, alkyl or alkoxy.

8. The compound of claim 1 which is

N- (3, 5-difluoro-phenyl) -N '- [3- (5-oxo-4, 5-dihydro- [1, 2, 4] oxadiazol-3-yl) -4' -trifluoromethyl-biphenyl-4-yl ] urea;

n- (4-chloro-3-trifluoromethyl-phenyl) -N '- {3- (5-oxo-4, 5-dihydro- [1, 2, 4] -oxadiazol-3-yl) -4' -trifluoromethyl-biphenyl-4-yl } urea;

n- (3-bromo-phenyl) -N '- {3- (5-oxo-4, 5-dihydro- [1, 2, 4] -oxadiazol-3-yl) -4' -trifluoromethyl-biphenyl-4-yl } urea;

n- (2-chloro-phenyl) -N '- {3- (5-oxo-4, 5-dihydro- [1, 2, 4] -oxadiazol-3-yl) -4' -trifluoromethyl-biphenyl-4-yl } urea;

or a pharmaceutically acceptable salt thereof.

9. A pharmaceutical composition comprising a therapeutically effective amount of a compound of any of claims 1-8, any of its isomers or any mixture of its isomers, or a pharmaceutically acceptable salt thereof, in association with at least one pharmaceutically acceptable carrier, excipient or diluent.

11. Use of a compound according to any one of claims 1-9, any of its isomers or any mixture of its isomers, or a pharmaceutically acceptable salt thereof, for the manufacture of a pharmaceutical composition for the treatment, prevention or alleviation of a disease or a disorder or a condition of a mammal, including a human, which disease, disorder or condition is the blockade of chloride channels or the modulation of BK_CaThe channel has a response.

12. The use of claim 11, wherein the pair blocks a chloride channel or modulates BK_CaThe channel responsive disease, disorder or condition is osteoclast-associated bone disease, osteoporosis, postmenopausal osteoporosis, secondary osteoporosis, osteolytic breast cancer bone metastasis, osteolytic cancer invasion, paget's disease of bone, cancer, metastatic cancer, prostate cancer, lung cancer, breast cancer, bladder cancer, kidney cancer, colon cancer, stomach cancer, pancreatic cancer, ovarian cancer, melanoma, hepatoma, sarcoma, lymphoma, exudative macular degeneration (AMD), age-related macular degeneration (AMD), retinopathy, diabetic retinopathy, proliferative diabetic retinopathy, Diabetic Macular Edema (DME), ischemic retinopathy, retinal vein or artery occlusion, retinopathy of prematurity, neovascular glaucoma, corneal neovascularization, ocular hypertension, open angle glaucoma, chronic open angle glaucoma, closed angle glaucoma, ciliary congestion due to angle-closure glaucoma, rheumatoid arthritis, psoriasis, sickle cell anemia, atherosclerosis, ischemia, reperfusion injury, hypertension, restenosis, arterial inflammation, myocardial ischemia, ischemic heart disease, airway hyperreactivity, pneumoconiosis, aluminosis, anthracosis, asbestosis, chalicosis, camelhair pneumoconiosis, iron pneumoconiosis, silicosis, tobacco pneumoconiosis, gossypium, sarcoidosis, berylliosis, emphysema, Acute Respiratory Distress Syndrome (ARDS), Acute Lung Injury (ALI), acute or chronic infectious pulmonary disease, Chronic Obstructive Pulmonary Disease (COPD), bronchitis, chronic bronchitis, asthmatic bronchitis, exacerbation of airway hyperreactivity or cystic fibrosis, cough, including chronic cough, exacerbation of airway hyperreactivity, pulmonary fibrosis, pulmonary hypertension, inflammatory pulmonary disease, or acute or chronic respiratory infectious disease, urinary incontinence, psychosis, epilepsy or pain.

13. Use of a compound according to any one of claims 1 to 9, any of its isomers or any mixture of its isomers, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for increasing blood-brain barrier permeability.

14. Methods of treating, preventing or ameliorating a disease or disorder or condition of a living animal body, including a human, which disorder, disease or condition is the blockade of chloride channels or modulation of BK_CaA channel responsive, comprising the step of administering to such a living animal body in need thereof a therapeutically effective amount of a compound according to any one of claims 1 to 9, any of its isomers or any mixture of its isomers, or a pharmaceutically acceptable salt thereof.