Nicotinamide derivates useful as p38 inhibitors

Description of the invention

The invention relates to a nicotinamide derivative, a pharmaceutical agent containing the derivative and the use of a nicotinamide derivative useful as a therapeutic agent, particularly as a p38 kinase inhibitor, for the treatment of conditions or diseases mediated by p38 kinase activity or mediated by cytokines produced by p38 kinase activity.

The nicotinamide derivative of the invention and its salts are novel p38 kinase inhibitors.

The invention relates to a nicotinamide derivative which is 6- (5-cyclopropylcarbamoyl-3-fluoro-2-methylphenyl) -N- (2,2-dimethylpropyl) nicotinamide or a pharmaceutically acceptable salt thereof.

The derivative 6- (5-cyclopropylcarbamoyl-3-fluoro-2-methylphenyl) -N- (2,2-dimethylpropyl) nicotinamide is preferred.

The invention also relates to a pharmaceutical composition containing the active ingredient in admixture with one or more pharmaceutically acceptable carriers, diluents or excipients, characterized in that the active ingredient is a nicotinamide derivative as defined above or a pharmaceutically acceptable salt thereof.

The invention also relates to a nicotinamide derivative as defined above or a pharmaceutically acceptable salt thereof for use in therapy.

The nicotinamide derivative according to the invention, 6- (5-cyclopropylcarbamoyl-3-fluoro-2-methylphenyl) -N- (2,2-dimethylpropyl) nicotinamide, is a compound of formula (I):

<img file="PL212910B1_D0001.tif" />

in which R.¹ is 2-dimethylethyl, R² is H, R.³ is CH3, R.⁴ is -CO-NH-cyclopropyl and X is F.

The nicotinamide derivatives may exist in stereoisomeric forms (e.g. they may contain one or more asymmetric carbon atoms or may exhibit cistrans isomerism). Thus, these derivatives may exist in the form of individual stereoisomers (enantiomers and diastereoisomers) and mixtures thereof, as well as individual isomers of nicotinamide derivatives as mixtures with their isomers in which one or more chiral centers are inverted. Likewise, it should be taken into account that the nicotinamide derivatives may exist in tautomeric forms.

Salts of the nicotinamide derivative according to the invention are within the scope of the invention and can, for example, be acid addition salts obtained by reacting an acid with a basic nitrogen atom present in the nicotinamide derivative.

Salts encompassed by the term "pharmaceutically acceptable salts" refer to the non-toxic salts of the nicotinamide derivative of the invention. Representative salts include the following: acetate, benzenesulfonate, benzoate, bicarbonate, bisulfate, bitartrate, borate, bromide, calcium edetate, camsylate, carbonate, chloride, clavulanate, citrate, dihydrochloride, edetate, edisylate, estolate, esilate, fumarate, gluceptan, gluconate, glutylarsorilate, glycolarsorilate , hydrabamine salt, hydrobromide, hydrochloride, hydroxynaphthoate, iodide, isethionate, lactate, lactobionate, laurate, malate, maleate, mandelate, mesylam, methylbromide, methylnitrate, methyl sulfate, monopotassium maleate, mucate, napsylate, nitrate, salt with N-methylglucamine, oxalate, pamoate (pamoate), palmitate, panthothenate, phosphate / diphosphate, polygalacturonate, potassium salt, salicylate, sodium salt, stearate tannate, basic acetate, acetate, , tartrate, theoclate, tosylate, triethyl iodide, trimethylammonium salt and valerate. Other non-pharmaceutically acceptable salts may be useful in the preparation of the nicotinamide derivative of the invention.

The nicotinamide derivative according to the invention, 6- (5-cyclopropylcarbamoyl-3-fluoro-2-methylphenyl) -N- (2,2-dimethylpropyl) nicotinamide, is a compound of formula (I):

<img file="PL212910B1_D0002.tif" />

which R.¹ is 2-dimethylethyl, R² is H, R.³ is CH3, R.⁴ is -CO-NH-cyclopropyl and X is F.

The nicotinamide derivatives of the invention can be prepared by a variety of methods, including conventional chemical methods. Exemplary general preparation methods are described below, and then specific nicotinamide derivatives of the invention were prepared in the examples given below.

Nicotinamide derivatives can be prepared by reacting compound (II)

<img file="PL212910B1_D0003.tif" />

in which R.¹ is 2-dimethylethyl, R² is H and W is halogen, in particular bromine or chlorine, with a compound of formula (III)

<img file="PL212910B1_D0004.tif" />

in which R.³ is CH3, R.⁴ is -CO-NH cyclopropyl and X is F, in the presence of a catalyst, e.g. tetrakis (triphenylphosphine) palladium.

The compound of formula (II) can be easily prepared from the corresponding acidic compound of formula (IV)

PL 212 910 B1

<img file="PL212910B1_D0005.tif" />

wherein W is as defined above, by converting the acid into an activated acid form, e.g. an acid chloride, by treatment with e.g. thionyl chloride and then reacting the activated acid obtained with an amine compound of formula (V)

<img file="PL212910B1_D0006.tif" />

in which R.¹ and r² are as defined above, under amide-forming conditions.

Suitable amide-forming conditions are known in the art and include treating a solution of the acid of formula (IV) or an activated form thereof in e.g. acetone or dichloromethane with an amine of formula (V) in the presence of potassium carbonate.

A compound of formula (III) can be prepared by reacting an acidic compound of formula (VII)

<img file="PL212910B1_D0007.tif" />

in which R.³, hal and X are as defined above, with bis (pinacolane) diboron, PdCl2dppf and potassium acetate in a solvent such as DMF followed by an amide formation by reaction with an amine compound of formula (V) as defined above.

Nicotinamide derivatives can also be prepared by reacting a compound of formula (VIII)

<img file="PL212910B1_D0008.tif" />

with a compound of formula (III) as defined above, followed by reaction of the acid so obtained with an amine of formula (V) as defined above, under amide-forming conditions.

Furthermore, nicotinamide derivatives can be prepared by reacting a compound (II) as defined above with a compound of formula (IX)

<img file="PL212910B1_D0009.tif" />

in which R.³, R⁴, X are as defined above in the presence of a catalyst, e.g., tetrakis (triphenylphosphine) palladium.

PL 212 910 B1

For example, the preparation of nicotinamide derivatives includes the reactions shown in Scheme 1 below.

Scheme 1

<img file="PL212910B1_D0010.tif" />

ABOUT

i. SOCI2.

ii. R⁸NH2, Na2CO3, DCM.

iii. NaH, n-BuLi, THF, (iPrO) 3B.

iV. SOCl2

v. R¹(CH2) R²NH, Na2CO3, DCM.

vi. NaHCO3, tetrakis (triphenylphosphine) palladium, propan-2-ol.

where hal, R.^{1 * *} and r² are as defined above, and R.⁸ means cyclopropyl.

The terms "halogen" or "halo" as used herein mean fluorine, chlorine, bromine or iodine. The preferred halogen atoms are fluorine, chlorine or bromine. A particularly preferred halogen atom is fluorine or chlorine.

As used herein, the term "optionally" means that the event (s) described thereafter may or may not occur, and includes case (s) that occur and that do not (- and).

While the nicotinamide derivatives of the invention can be administered as new chemicals, the nicotinamide derivatives are usually administered in the form of pharmaceuticals. Thus, in another embodiment, the invention relates to a pharmaceutical composition comprising a nicotinamide derivative in admixture with one or more pharmaceutically acceptable carriers, diluents or excipients.

The nicotinamide derivatives may be formulated for administration by any method. They may e.g. be formulated for topical administration or administration by inhalation or, more preferably, for oral, transdermal or parenteral administration. The pharmaceutical agent may be present at 6

A form which can cause a controlled release of nicotinamide derivatives. Oral administration of the formulation is a particularly preferred mode of administration, and a suitable formulation.

For oral administration, a pharmaceutical may be in the form of e.g. tablets (including sublingual tablets) and administered as tablets and capsules (being time release and sustained release formulations), pills, powders, granules, elixirs, tinctures, emulsions, solutions. , syrups or suspensions prepared by conventional methods with acceptable excipients.

For example, for oral administration in the form of a tablet or capsule, the active drug component can be combined with an oral, non-toxic pharmaceutically acceptable inert carrier such as ethanol, glycerin, water, etc. Powders are prepared by grinding the compound to an appropriately selected particle size and mixing with a similarly particulate. a pharmaceutical carrier such as an edible carbohydrate such as, for example, starch or mannitol. A flavoring, preservative, dispersing, and coloring agent may also be present.

Capsules can be made by preparing a powder mixture as described above, and filling the prepared gelatin sheaths. Glidants and lubricants such as colloidal silica, talc, magnesium stearate, calcium stearate, or solid polyethylene glycol may be added to the powder mixture prior to the filling operation. A disintegrating or solubilizing agent such as agar, calcium carbonate, or sodium carbonate may be added to improve the availability of the drug when the capsule is swallowed.

Moreover, when desired or necessary, suitable binders, lubricants, disintegrating agents and coloring agents can be incorporated into the mixture. Suitable binders include starch, gelatin, natural sugars such as glucose or beta-lactose, corn sweeteners, natural and synthetic gums such as acacia, tragacanth or sodium alginate, carboxymethylcellulose, polyethylene glycol, waxes etc. Lubricants used in these dosage forms include sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride, and the like. Disintegrants include, without limitation, starch, methylcellulose, agar, bentonite, xanthan gum, and the like. Tablets are formulated e.g. by preparing a powder mixture, granulating or spheronizing, adding a lubricant and disintegrant, and pressing into tablets. The powder mixture is prepared by mixing the suitably comminuted compound with a diluent or vehicle as described above and optionally with a binder such as carboxymethyl cellulose, alginate, gelatin or polyvinylpyrrolidone, a dissolution retarding agent such as paraffin, a resorption accelerator such as a quaternary salt, and / or an absorbent agent such as bentonite, kaolin or dicalcium phosphate. The powder mixture can be granulated by wetting with a binder such as syrup, starch paste, acacia or solutions of cellulosic or polymeric substances and extruding through a screen. As an alternative to granulating, the powder mixture can be run through the tablet machine and imperfectly formed slugs broken into granules are obtained. The granules may be lubricated to prevent sticking to the tablet forming dies by the addition of stearic acid, stearate, talc or mineral oil. The lubricated mixture is then compressed into tablets. The nicotinamide derivatives of the invention can also be combined with a free flowing inert carrier and compressed into tablets directly without going through the granulating or spheronizing steps. A transparent or opaque protective coating may be applied, consisting of a seal coat of shellac, a sugar coating or a polymeric material and a glossy wax coating. Dyes may be added to these coatings to distinguish between different dosage units.

Oral fluids such as solution, syrup, and elixirs can be prepared in dosage unit form so that a given amount contains a pre-measured amount of the compound. Syrups can be prepared by dissolving the compound in an aqueous solution of appropriate flavor and aroma, while elixirs are prepared using non-toxic alcoholic vehicles. Suspensions can be formulated by dispersing the compound in a non-toxic vehicle. Solubilizers and emulsifiers such as ethoxylated isostearyl alcohols and polyoxyethylene sorbitan ethers, preservatives, flavoring additives such as peppermint oil or saccharin, etc. may also be added.

Where appropriate, dosage unit formulations for oral administration may be microencapsulated. A sustained or sustained release formulation may also be prepared, e.g. by coating or embedding the particulate material in polymers, wax or the like.

The nicotinamide derivatives of the invention can also be administered in the form of liposomal delivery systems such as small unilamellar vesicles, large unilamellar vesicles

PL 212 910 B1 and multi-layered vesicles. Liposomes can be formed from various phospholipids such as cholesterol, stearylamine, or phosphatidylcholine.

The nicotinamide derivatives of the invention can also be administered in the form of liposomal emulsion delivery systems, such as small unilayered vesicles, large unilayered vesicles, and multilayer vesicles. Liposomes can also be formed from various phospholipids such as cholesterol, stearylamine, or phosphatidylcholines.

The nicotinamide derivatives of the invention can also be delivered using monoclonal antibodies as individual carriers to which the compound molecules are attached. The nicotinamide derivatives according to the invention can also be combined with soluble polymers as target drug carriers. Such polymers may include polyvinylpyrrolidone, pyran copolymer, polyhydroxypropyl methacryl amidophenol, polyhydroxyethylaspartamidophenol or polyethyleneoxydopolylysine substituted with palmitoyl residues. In addition, the nicotinamide derivatives of the invention may be attached to a class of biodegradable polymers useful in achieving controlled drug release, such as e.g. polylactic acid, polypsilone caprolactone, polyhydroxybutyric acid, polyorthoesters, polyacetals, polydihydropyranes, polycyanoacrylates, and cross-linked or amphipathic block copolymers of hydrogels.

The pharmaceutical composition according to the invention usually contains 0.1-99.5%, preferably 0.5-90%, of a nicotinamide derivative in combination with a pharmaceutically acceptable carrier.

Likewise, the agent may also be administered intranasally, ophthalmically, in the ear, rectally, topically, intravenously (both high dose and infusion), intraperitoneal, intraarticular, subcutaneous or intramuscular, inhaled or insufflated, all forms used are known in the art. pharmaceutical professionals.

For transdermal administration, the pharmaceutical agent may be administered in the form of a transdermal patch, such as a transdermal iontophoretic patch.

For parenteral administration, the pharmaceutical agent can be administered by injection or continuous infusion (e.g., intravenously, intravascularly, or subcutaneously). 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. For injection, the agents may be in the form of a unit dose preparation or a multi-dose preparation, preferably with an added preservative. Alternatively, for parenteral administration, the active ingredient may be in powder form for constitution with a suitable vehicle.

The nicotinamide derivatives of the invention may also be formulated as a depot preparation. Such long-acting formulations can be administered by implantation (e.g., subcutaneously or intramuscularly) or by intramuscular injection. Thus, for example, the nicotinamide derivatives of the invention may be formulated with suitable polymeric or hydrophobic materials (e.g. as an emulsion in an acceptable oil) or ion exchange resins, or as sparingly soluble derivatives, e.g. as sparingly soluble salts.

Alternatively, the agent may be formulated for topical application, e.g., in the form of ointments, creams, lotions, eye ointments, eye drops, ear drops, mouth washes, impregnated dressings and sutures, and aerosols, and may contain suitable conventional additives such as including, e.g., preservatives, solvents to facilitate drug penetration, and emollients in ointments and creams. Such topical preparations may also contain compatible conventional carriers, e.g. cream or ointment bases, and ethanol or oleyl alcohol for lotions. Such carriers can make up about 1-98 wt.%. preparation; they usually constitute up to about 80 wt.%. preparation.

For administration by inhalation, the nicotinamide derivatives of the invention are typically provided in the form of a pressurized aerosol spray formulation or a nebulizer using a suitable propellant e.g. In the case of a pressurized aerosol, the dosage unit may be metered by a valve to deliver a metered amount. Capsules and cartridges of e.g. gelatin for use in an inhaler or insufflator may be formulated containing a powder mix of the nicotinamide derivative of the invention and a suitable powder base such as lactose or starch.

Pharmaceuticals are usually administered in an amount effective to treat or prevent the particular condition or conditions. Initial human dosing is combined with clinical monitoring for symptoms such as symptoms for the selected condition. Generally, the agents are administered in an amount of active ingredient of at least about 100 µg / kg body weight. In most cases, they are given in either or

In a number of more doses, no more than 20 mg / kg body weight per day. Preferably, in most instances, the dose is between about 100 µg / kg - 5 mg / kg body weight per day. For administration to individual mammals, and especially humans, it is expected that the daily dose level of the active ingredient is 0.1 mg / kg - 10 mg / kg, and usually about 1 mg / kg. It should be understood that the optimal dose will be determined by the usual methods for each treatment regimen and indication, taking into account the indication, its severity, route of administration, complications, etc. The physician in each case will determine the actual dose which will be most appropriate for the subject and will be vary with the age, weight and reaction of a particular individual. The effectiveness of the selected actual dose can be easily determined e.g. by measuring clinical symptoms or the usual anti-inflammatory indication after administration of a selected dose. The above dosing methods are exemplary of the average case. These may, of course, include individual cases where higher or lower dosage ranges are recommended, and such are included within the scope of the invention. For disease states or conditions as treated in accordance with the invention, maintaining constant daily levels in a subject for an extended period of time, e.g. in maintenance mode, may be particularly advantageous.

In another embodiment, the invention relates to a nicotinamide derivative for use in therapy.

The nicotinamide derivatives of the invention are generally p38 serine / threonine kinase inhibitors and therefore also inhibitors of cytokine production which is mediated by p38 kinase. The meaning of the term "p38 serine / threonine kinase inhibitors" includes those compounds that interfere with the ability of p38 to transfer a phosphate group from ATP to a protein substrate according to the assay described below.

It should be appreciated that the nicotinamide derivatives of the invention may be selective for one or more isoforms of p38, e.g. ρ38α, ρ38β, ρ38γ and / or ρ38δ. In one embodiment, the nicotinamide derivatives of the invention selectively inhibit the p38a isoform. In another embodiment, the nicotinamide derivatives of the invention selectively inhibit the ρ38β isoform. In a further embodiment, the nicotinamide derivatives of the invention selectively inhibit the isoforms ρ38α and ρ38β. Assays for determining the selectivity of compounds for the p38 isoforms are described in e.g. WO 99/61426, WO 00/71535 and WO 02/46158.

It is known that p38 kinase activity may be increased (locally or through the body), p38 kinase may be incorrectly temporally active or expressed, p38 kinase may be expressed or active at the wrong site, p38 kinase may be constitutively expressed, or expression of p38 kinase may be variable ; similarly, cytokine production mediated by p38 kinase activity may occur at inappropriate times, inappropriate sites, or may occur at deleteriously high levels.

Thus, the invention enables the treatment of a condition or disease mediated by p38 kinase activity, or mediated by cytokines produced by p38 kinase activity in a subject, by administering to the subject a therapeutically effective amount of a nicotinamide derivative. A compound can be administered in a single or polymorphic crystalline form or forms, amorphous form, as a single enantiomer, a racemic mixture, a single stereoisomer, a mixture of stereoisomers, a single diastereoisomer, or a mixture of diastereoisomers.

The invention also enables the inhibition of cytokine production mediated by p38 kinase activity in a subject, e.g. a human, by administering to the subject a therapeutic or cytokine inhibitory amount of a nicotinamide derivative of the invention. A compound can be administered in a single or polymorphic crystalline form or forms, amorphous form, as a single enantiomer, a racemic mixture, a single stereoisomer, a mixture of stereoisomers, a single diastereoisomer, or a mixture of diastereoisomers.

The conditions described are treated by the provision of a therapeutically effective amount of a nicotinamide derivative according to the invention. By "therapeutically effective amount" is meant an amount of a compound that alleviates a symptom or reduces a symptom, an amount of a cytokine reducing compound, an amount of a cytokine inhibiting compound, a kinase regulating amount of a compound, and / or a kinase inhibiting amount of a compound. Such amounts can be readily determined by conventional methods such as measuring cytokine levels or observing the improvement of clinical symptoms. For example, a physician may monitor acceptable measurement results for an anti-inflammatory treatment.

The nicotinamide derivatives of the invention can be administered to any subject in need of inhibition or regulation of p38 kinase or in need of inhibition or regulation of p38 kinase-mediated cytokine production. In particular, the compounds can be administered to mammals. Such mammals can be, for example, horses, cows, sheep, pigs, mice, dogs, cats, primates such as chimpanzees, gorillas, rhesus monkeys and preferably humans.

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Thus, the invention enables the treatment or reduction of symptoms in humans or animals suffering from e.g. rheumatoid arthritis, osteoarthritis, asthma, psoriasis, eczema, allergic rhinitis, allergic conjunctivitis, adult respiratory distress syndrome, chronic pneumonia, chronic obstructive pulmonary disease, chronic heart failure, silicosis, endotoxemia, toxic shock syndrome , inflammatory bowel disease, tuberculosis, atherosclerosis, neurodegenerative diseases, Alzheimer's disease, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis, epilepsy, multiple sclerosis, aneurysm, stroke, irritable bowel syndrome, muscle degeneration, bone resorption diseases, osteoporosis, diabetes, reperfusion injury, graft versus host reaction, allograft rejection, sepsis, systemic wasting, secondary wasting infection or cancer, wasting secondary from acquired immune deficiency syndrome (AIDS), malaria, leprosy, infectious arthritis, leishmaniasis, Lyme disease, glomerulonephritis, gout, psoriatic arthritis, Reiter's syndrome, traumatic arthritis, rubella arthritis, Crohn's disease, ulcerative colitis, acute synovitis, gouty arthritis, spondylitis, and arthritis inflammatory e.g. herniated / ruptured / prolapsed intervertebral disc syndrome, bursitis, tendinitis, tenosynovitis, fibromyalgic syndrome and other inflammatory conditions associated with ligament tear injury without dislocation and regional musculoskeletal injury, pain, e.g. which is associated with inflammation and / or trauma, bone marbling, restenosis, thrombosis, angiogenesis, cancer, including breast cancer, colon cancer, lung cancer, or prostate cancer, by administering to a subject a therapeutically effective amount of a nicotinamide derivative.

In particular, the invention enables the treatment of a human or animal suffering from rheumatoid arthritis, asthma, psoriasis, chronic pneumonia, chronic obstructive pulmonary disease, chronic heart failure, systemic wasting, glomerulonephritis, Crohn's disease, neurodegenerative disease, Alzheimer's disease, Parkinson's disease, epilepsy and cancer, including breast cancer, colon cancer, lung cancer and prostate cancer by administering to a subject a therapeutically effective amount of a nicotinamide derivative.

In particular, the invention also enables the treatment of a human or animal suffering from rheumatoid arthritis, asthma, psoriasis, chronic pneumonia, chronic obstructive pulmonary disease, chronic heart failure, systemic wasting, glomerulonephritis, Crohn's disease and cancer, including breast cancer, cancer colon, lung cancer, and prostate cancer, by administering to a subject a therapeutically effective amount of a nicotinamide derivative.

In particular, the invention also enables the treatment of a human or animal suffering from rheumatoid arthritis, asthma, chronic pneumonia, chronic obstructive pulmonary disease, neurodegenerative diseases, Alzheimer's disease, Parkinson's disease and epilepsy by administering to the subject a therapeutically effective amount of a nicotinamide derivative.

The invention enables the treatment of a human or animal suffering from any type of pain, including chronic pain, rapid onset of analgesia, neuromuscular pain, headache, cancer pain, acute and chronic inflammatory pain associated with osteoarthritis and rheumatoid arthritis. arthritis, postoperative inflammatory pain, neuropathic pain, diabetic neuropathy, trigeminal neuralgia, hepatitis neuralgia, inflammatory neuropathies and migraine pain, by administering to a subject a therapeutically effective amount of a nicotinamide derivative or a pharmaceutically acceptable salt thereof.

The nicotinamide derivatives can be used alone or in combination with other therapeutic agents to treat the above-mentioned conditions. In particular, in the treatment of rheumatoid arthritis, combination with other chemotherapeutic agents or antibodies is envisaged. The combination therapy thus comprises the administration of at least one nicotinamide derivative and at least one other pharmaceutically active agent. The nicotinamide derivative (s) and the other pharmaceutically active agent (s) may be administered together or separately and, when administered separately, the administration may be separate or sequential in any order. The amounts of the nicotinamide derivative (s) and the other pharmaceutically active agent (s) and the appropriate timing of administration will be selected to achieve the desired combined therapeutic effect. Examples of other pharmaceutically active agents that can be used in combination with compounds of formula (I) for the treatment of rheumatoid arthritis are: immunosuppressants such as amtolmetin guacil, misoribine and rimexolone; anti-TNFα agents such

Such as etanercept, infliximab, diacerein; tyrosine kinase inhibitors such as leflunomide; kallikrein antagonists such as subreum; interleukin 11 agonists such as oprerevekine; interferon beta 1 agonists; hyaluronic acid agonists such as NRD-101 (Aventis); interleukin 1 receptor antagonists such as anakinra; CD8 antagonists such as amiprilose hydrochloride; β-amyloid precursor protein antagonists such as rheumacone; matrix metalloprotease inhibitors such as cypemastat and other disease-modifying anti-rheumatic drugs (LAMCh) such as methotrexate, sulfasalazine, cyclosporin A, hydroxychoroquine, auranofin, aurothioglucose, gold sodium thiomalate and penicillamine.

Examples

The following examples illustrate the invention. The reagents are either commercially available or are prepared according to methods described in the literature.

LCMS was performed on a column (3.3 cm x 4.6 mm ID, 3 μm ABZ + PLUS), flow rate 3 ml / min, injection volume 5 μl, at room temperature and in the UV detection range at 215-330 nm.

General method A.

6-Bromonicotinic acid (100 mg, 0.5 mmol) was heated at 95 ° C in thionyl chloride (0.63 µm) for 2 hours. Excess thionyl chloride was evaporated in vacuo and the residue was dissolved in DCM (2 ml). To this solution were added amine (0.5 mmol) and sodium carbonate (100 mg), and the reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was filtered and the residue was washed with DCM. The combined filtrate and washes were concentrated to dryness to give the desired 6-oronicotinamide.

<td>Relationship</td><td>Amine</td><td>MH +</td><td>Time retention (minutes)</td>

<td>Intermediate 1: 6-Chloro-N- (2,2-dimethylpropyl)) nicotinamide</td><td>2,2-dimethylpropylamine</td><td> 227</td><td> 2,82</td>

Intermediate 2. N-Cyc | opropy | o-5-f | uoro-4-methyl | o-3- (4,4,5,5-tetrameth | o [1.3,2] dioxaboro | an-2y | o ) benzamide

3-Bromo-N-cyc | ops | o-5-f | uoro-4-methylbenzamide (intermediate 3, 900 mg), bispinaco | anodiborium (4.5 g), potassium acetate (2.1 g) and PdC [Beta] 2dppf (75 mg) was stirred in DMF (40 ml) and heated at 100 [deg.] C for 18 hours. The cooled reaction mixture was absorbed onto silica and loaded onto an SPE (Si 2 X 10 g). The SPE was subjected to a gradient elution with ethyl acetate / cyclohexane (0-6.25% ethyl acetate). The solvent was evaporated from the fractions containing the product in vacuo and the residue was recrystallized from cyclohexane to give N-cycloprops | o-5-f | uoro-4-methyl-3- (4,4,5,5-tetrameters | o- [1,3,2] dioxaboro | an-2-y] benzamide (260 mg). LCMS: MH⁺320, retention time 3.39 minutes.

Intermediate 3. 3-Bromo-N-cyc | fuels | o-5-f | uoro-4-methyl | obenzamide

3-f | uoro-4-methyl | benzoic acid (462 mg, 3.0 mmol) was added with stirring to a mixture of bromine (2.31 m |, 45 mmol) and iron powder (252 mg, 4 5 mmo | a) under nitrogen atmosphere. The reaction mixture was stirred at 20 ° C for 4 hours and then allowed to stand for 16 hours. Sodium thiosulfate solution (200 ml) was added and the product was extracted with ethyl acetate (3 x 150 ml). The organic extracts were combined and evaporated in vacuo. The crude product (mixture of isomers) was dissolved in dimethyl oformamide (7 ml). Cyc | opropylamine (208 μ |, 3.0 mmo | e), HOBT (405 mg, 3.0 mmo | e), 1- (3-dimethyl | oaminopropy | o) hydrochloride were added to the solution with stirring. 3-ethylcarbodiimide (575 mg, 3.0 mmol) and DIPEA (525 µl, 3.0 mmol). The reaction mixture was stirred for 5 hours at 20 ° C. The solvent was removed in vacuo and the residue was partitioned between ethyl acetate and water. The combined organic extracts were washed sequentially with aqueous sodium bicarbonate and hydrochloric acid (0.5M), then dried (magnesium sulfate). Ethyl acetate was evaporated in vacuo and the residue was purified by biotage chromatography on silica eluting with cyclohexane: ethyl acetate (6: 1) to give 3-bromo-N-cyclopropyl-5-fluoro-4-methylbenzamide (359mg, 44%). NMR: δΗ - CDCl₃ 7.68, (1H,

S), 7.39, (1H, d), 6.19, (1H, bs), 2.88, (1H, m), 2.36, (3H, d), 0.88, (2H, m), 0.63, (2H, m). LCMS: MH⁺ 272.

Intermediate 4. {5 - [(cyc | oppy | oamino) carbony | o] -3-f | uoro-2-methyl | ofeny | o} boronic acid

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N-Cyclopropyl-5-fluoro-3-iodo-4-methylbenzamide (intermediate 5.5 g) in THF (75 ml) was cooled to 0 ° C and sodium hydride (60%, 1.23 g) was added batchwise overnight. 10 minutes. After gas evolution ceased, the reaction mixture was cooled to -75 ° C and n-butyllithium (1.6M in hexanes, 20 mL) was added over 25 minutes keeping the temperature &lt;-70 ° C. To the reaction mixture was added triisopropyl borate (8 ml) over 10 minutes and the reaction mixture was stirred at -70 ° C for 4 hours. The reaction mixture was quenched with water (20 mL) and warmed to 5 ° C. The reaction mixture was concentrated in vacuo and the residue was partitioned between saturated ammonium chloride and ethyl acetate. The organic phase was washed with saturated ammonium chloride, brine, dried (sodium sulfate) and concentrated to dryness in vacuo. The residue was dissolved in DCM / ethyl acetate and purified by silica column chromatography eluting with a gradient of ethyl acetate / DCM (5-100% ethyl acetate) and then methanol. The product fractions were combined and the solvent evaporated in vacuo to give {5 - [(cyclpropylamino) carbonyl] -3-fluoro-2-methylphenyl} boronic acid. LCMS MH⁺ 238, retention time 2.19 minutes.

Intermediate 5. N-Cyclopropyl-5-fluoro-3-iodo-4-methylbenzamide

N-iodosuccinimide (22.5 g) was added portionwise to a solution of 3-fluoro-4-methylbenzoic acid (15.4 g) in trifluoromethanesulfonic acid (100 ml) at 0 ° C for 3 hours and the reaction was warmed to room temperature for 3 hours. night. The reaction mixture was poured into ice water (400 ml) and the precipitate was filtered off and washed with water. The solid residue was dissolved in ethyl acetate, the solution was washed with aqueous sodium thiosulfate (x2) then brine, dried (magnesium sulfate) and the solvent was evaporated in vacuo. The residue was mixed with thionyl chloride (30 ml) and heated at 100 ° C for 2.5 hours. Excess thionyl chloride was removed from the cooled reaction mixture in vacuo and the residue was dissolved in DCM (100 ml). Sodium carbonate (25 g) and cyclopropylamine (13 ml) were added to the solution, and the reaction mixture was stirred at room temperature for 72 hours. The reaction mixture was filtered and the residue was washed with DCM and ethyl acetate. The solvent was evaporated from the filtrate combined with the washing liquid under vacuum. The residue was absorbed onto silica and subjected to flash column chromatography on silica eluted with a gradient of ethyl acetate / cyclohexane (22-28% ethyl acetate). The appropriate fractions were concentrated to dryness in vacuo to give N-cyclopropyl-5-fluoro-3-iodo-4-methylbenzamide. LCMS; MH⁺ 320, retention time 3.16 minutes.

Intermediate 6. 6- {5 - [(cyclopropylamino) carbonyl] -3-fluoro-2-methylphenyl} -nicotinic acid

N-Cyclopropyl-5-fluoro-4-methyl-3- (4,4,5,5-tetramethyl- [1,3,2] dioxaborolan-2-yl) benzamide (3.2 g), methyl 6-chloronicotinate (1.73 g), tetrakis (triphenylphosphine) palladium (210 mg) and aqueous sodium bicarbonate solution (1M, 30 ml) were stirred in propan-2-ol (100 ml) and heated at 90 ° C for 18 hours. The reaction mixture was cooled and propan-2-ol was removed in vacuo. The residue was partitioned between ethyl acetate and an aqueous sodium bicarbonate solution (1M). The aqueous phase was acidified with hydrochloric acid (2N) and extracted with ethyl acetate (x2). The organic extracts were washed with brine, dried (magnesium sulfate) and concentrated to dryness in vacuo. The resulting foam was triturated with ether to afford 6- {5 - [(cyclopropylamino) carbonyl] -3-fluoro-2-methylphenyl} nicotinic acid as a solid. LCMS: MH⁺ 315, retention time 2.87 minutes.

General method C.

6-Chloronicotinamide (25 mg), N-cyclopropyl-5-fluoro-4-methyl-3- (4,4,5,5-tetramethyl [1,3,2] -dioxaborolan-2-yl) benzamide (intermediate 2.15 mg), tetrakis (triphenylphosphine) palladium (2 mg) and aqueous sodium bicarbonate solution (1M, 0.5 ml) were stirred in propan-2-ol (2 ml) and refluxed for 18 hours. Propan-2-ol was evaporated and the residue was diluted with ethyl acetate / cyclohexane (1: 2). The solution was loaded onto an SPE (Si, 2 g) and eluted with ethyl acetate / cyclohexane (1: 2) and then ethyl acetate. The solvent was evaporated from the ethyl acetate fractions and the residue was triturated with ether to afford the desired product as a white solid.

Example 1 can also be prepared using {5 - [(cyclopropylamino) carbonyl] -3-fluoro-2-methylphenyl} boronic acid (interm. 4) instead of intermediate 2.

PL 212 910 B1

<td>Relationship</td><td colspan="2">Structure</td><td>6-Chloronicotinamide</td><td>MH +</td><td>Time retention (minutes)</td>

<td>Example L.</td><td></td><td>H Cft</td><td>6-chloro-N- (2,2-dimeth-</td><td> 384</td><td> 3,01</td>

<td>6- (5-cyclopropylcar-</td><td>ck</td><td>i \&gt; CH,</td><td>you! opropyl)) nicotine-</td><td></td><td></td>

<td>bamoyl-3-fluoro-2-</td><td></td><td>ch₃</td><td>amide</td><td></td><td></td>

<td>methylphenyl) -N- (2.2-</td><td></td><td></td><td>(interm. 1)</td><td></td><td></td>

<td>dimethylpropyl) ni-</td><td></td><td>j)</td><td></td><td></td><td></td>

<td>catinamide</td><td></td><td>Ti 'i¹</td><td></td><td></td><td></td>

<td></td><td></td><td></td><td></td><td></td><td></td>

Shortcuts

DCM dichloromethane

DIPEA N, N-diisopropylethylamine

DMF dimethylformamide

DMSO dimethyl sulfoxide

HOBT 1-hydroxybenzotriazole hydrate bond-elut SPE (solid phase extraction column

The activity of the nicotinamide derivatives of the invention as p38 inhibitors can be determined in the following test.

P38 kinase assay

The peptide substrate used in this p38 assay was biotin-IPTSPITTTYFFFRRR-amide. The p38 and MEK6 proteins were purified to homogeneity from E. coli expression systems. Fusion proteins were labeled N-terminally with glutathione-S-transferase (GST). Maximum activation was achieved by incubating 20 μΐ of the reaction mixture of 30 nM MEK6 protein and 120 nM p38 protein in the presence of 1.5 μΜ peptide and 10 mM Mg (CH3CO2) 2 in 100 mM HEPES, pH 7.5, added to 15 μΐ of mixture 1, 5 μΜ ATP with 0.08 μθ [g-³³P] ATP, with or without 15 µΐ inhibitor in 6% DMSO.

Controls were reacted in the presence (negative control) or without (positive control) of 50 mM EDTA. Reactions were carried out for 60 minutes at room temperature, stopped by the addition of 50 µM 250 mM EDTA and mixed with 150 µL SPA beads with streptavidin (Amersham) at 0.5 mg / reaction. The white U-shaped plates, Dynatech Microfluor, were closed and the beads were allowed to settle overnight. Platelet measurements were taken on the Packard TopCount within 60 seconds. IC50 values ​​were obtained by fitting the raw data to the equation% I = 100 * (1- (I-C2) / (C1-C2)), where I is the background CPM (pulses / minute), C1 is the positive control value, and C2 for the negative control.

The α p38 α p38 fluorescence polarization method was prepared in the laboratory. The SB4777790-R ligand was diluted in HEPES containing MgCl2, CHAPS, DTT and DMSO. The mixture was added to blank wells of a Black NUNC 384-well plate. Α p38 was added to this ligand mixture, and then added to the remaining wells of a 384-well plate containing controls and compounds. The plates were read on the LJL Analyst and the fluorescence anisotropy was used to calculate compound inhibition.

Results

The compounds described in the examples were tested as described above and found to exhibit IC50 values ​​&lt;10 μΜ.