Patent Application
20120196884
The present invention relates to sulphonamide derivatives, with a urea moiety. The invention also relates to the use of the derivatives as inhibitors of collagen receptor integrins, especially α2β1 integrin inhibitors and more precisely α2β1 integrin I-domain inhibitors, e.g. in connection with diseases and medical conditions that involve the action of cells and platelets expressing collagen receptors, their use as a medicament, e.g. for the treatment of thrombosis, inflammation, cancer and vascular diseases, pharmaceutical compositions containing them and a process for preparing them.
The integrins are a large family of cell adhesion receptors, which mediate anchoring of all human cells to the surrounding extracellular matrix. In addition integrins participate in various other cellular functions, including cell division, differentiation, migration and survival. The human integrin gene family contains 18 alpha integrin genes and 8 beta integrin genes, which encode the corresponding alpha and beta subunits. One alpha and one beta subunit is needed for each functional cell surface receptor. Thus, 24 different alpha-beta combinations exist in human cells. Nine of the alpha subunits contain a specific “inserted” I-domain, which is responsible for ligand recognition and binding. Four of the α I-domain containing integrin subunits, namely α1, α2, α10 and α11, are the main cellular receptors of collagens. Each one of these four alpha subunits forms a heterodimer with beta1 subunit. Thus the collagen receptor integrins are α1β1, α2β1, α10β1 and α11β1 (Reviewed in White et al., Int J Biochem Cell Biol, 2004, 36:1405-1410). Collagens are the largest family of extracellular matrix proteins, composed of at least 27 different collagen subtypes (collagens I-XXVII).
Integrin α2β1 is expressed on epithelial cells, platelets, inflammatory cells and many mesenchymal cells, including endothelial cell, fibroblasts, osteoblasts and chondroblasts (Reviewed in White et al., supra). Epidemiological evidence connect high expression levels of α2β1 on platelets to increased risk of myocardial infarction and cerebrovascular stroke (Santoso et al., Blood, 1999, Carlsson et al., Blood. 1999, 93:3583-3586), diabetic retinopathy (Matsubara et al., Blood. 2000, 95:1560-1564) and retinal vein occlusion (Dodson et al., Eye. 2003, 17:772-777). Evidence from animal models supports the proposed role of α2β1 in thrombosis. Integrin α2β1 is also overexpressed in cancers such as invasive prostate cancer, melanoma, gastric cancer and ovary cancer. These observations connect α2β1 integrin to cancer invasion and metastasis. Moreover, cancer-related angiogenesis can be partially inhibited by anti-α2 function blocking antibodies (Senger et al., Proc. Natl. Acad. Sci. U.S.A., 1997, 94:13612-13617). In addition inflammatory cells are partially dependent on α2β1 function during inflammatory process (de Fougerolles et al., J. Clin. Invest., 2000, 105:721-729; Edelson et al., Blood, 2004, 103:2214-2220). Based on the tissue distribution and experimental evidence α2β1 integrin may be important in inflammation, fibrosis, bone fracture healing and cancer angiogenesis (White et al., supra), while all four collagen receptor integrins may participate in the regulation of bone and cartilage metabolism.
The strong evidence indicating the involvement of collagen receptors in various pathological processes has made them potential targets of drug development. Function blocking antibodies against α1 or α2 subunits have been effective in several animal models including models for inflammatory diseases and cancer angiogenesis. Synthetic peptide inhibitors as well as snake venom peptides blocking the function of α1β1 and α2β1 have been described. (Eble, Curr Pharm Design 2005, 11:867-880). International Patent Publication WO 99/02551 discloses one small molecule drug candidate that regulates the expression of α2β1 but does not actually bind to the integrin.
Publication EP 1 258 252 A1 describes certain N-indolyl-, N-quinolinyl-, N-isoquinolinyl- and N-coumarinyl-arylsulphonamides, which are stated to be integrin expression inhibitors. Said publication does not specifically disclose the compounds of the present invention. Further, said known compounds differ from the compounds now described with respect to their properties and the mechanism of function. The compounds of the present invention are not integrin expression suppressors.
Publication EP 0 472 053 B1 discloses sulphonamides having anti-tumor activity. The compounds specifically described in said publication do not fall within the definition of the compound group of the present invention.
Publication Izvestiya Aakademii Nauk SSSR, Seriya Khimicheskaya (1981), (6), Kravtsov, D. N. et al., pp. 1259-1264 discloses sulphonamides, which are structurally closely related to the compounds now described but which do not fall within the definition of the compound group of the present invention. The field of use of the known compounds is totally different from that of the present invention.
Publication WO 2004/005278 discloses bisarylsulphonamides and their use in cancer therapy.
Publication WO 2007/034035 discloses a group of sulphonamide derivatives, which are useful as inhibitors of collagen receptor integrins.
Also publications WO 00/17159 A1, U.S. Pat. No. 5,939,431 A and U.S. Pat. No. 5,780,483 A disclose amylsulphonamides, which, however, are not described to be useful as inhibitors of collagen receptor integrins.
There is a constant need to develop new compounds, which are potentially useful in treatment of thrombosis, cancer, fibrosis and inflammation through inhibiting collagen receptor integrins. It is essential that collagen receptor integrin inhibitors have high activity, excellent bioactivity and good solubility and/or pharmacological properties.
It has now surprisingly been found that the sulphonamide derivatives with a urea moiety of the present invention are potent inhibitors for collagen receptor integrins, especially α2β1 integrin, and may be used in the treatment of human diseases, such as thrombosis, cancer, fibrosis, inflammation and vascular diseases. The derivatives according to the invention may also be used in diagnostic methods both in vitro and in vivo.
The present invention relates to a sulphonamide derivative of formula (I) or (I′) or a physiologically acceptable salt thereof,
where
R1 is H, C1-6-alkyl optionally substituted with one or two hydroxyl groups, C2-6-alkenyl optionally substituted with one or two hydroxyl groups, R′R″N—C1-6-alkyl-, C1-6-alkanoyl, R′OOC—C1-6-alkyl-, R′OOC—C1-6-alkoxy- or C1-6-alkoxy-C1-6-alkyl-;
R2 and R2′ are independently selected from H and C1-6-alkyl;
L is absent or a linker, which is a linear or a branched hydrocarbon chain with 1-6 carbon atoms;
X is a 5- or 6-membered aromatic ring with 0-2 heteroatoms selected from N, O and S and optionally substituted with R3;
R3 is OH, C1-6-alkyl optionally substituted with one or two hydroxyl groups, C2-6-alkenyl optionally substituted with one or two hydroxyl groups, halo-C1-6-alkyl, halo-C1-6-alkoxy, cyclo-C3-6-alkyl, C1-6-alkoxy, C1-6-alkanoyl, R′OOC—C1-6-alkyl-, R′OOC—C1-6-alkoxy-, —NO2, —CN, NC—C1-6-alkyl-, halogen, R″R′N—C1-6-alkyl-, R″R′N—C1-6-alkoxy-, R″-C(O)—NR′—C1-6-alkyl-, R″R′N—C(O)—C1-6-alkyl, R″-C(O)—NR′—C1-6-alkoxy-, R″R′N—C(O)—C1-6-alkoxy, —NR′R″, —NR′—C(O)—R″, —C(O)—NHR′, C1-6-alkoxy-C1-6-alkyl- or C1-6-alkoxy-C1-6-alkoxy-;
alternatively R2 and R3 form together a moiety selected from one of the following:
Ar1 is a 5- or 6-membered saturated or unsaturated ring with 0 to 2 heteroatoms selected independently from N, O and S and optionally substituted with one or more groups selected from C1-6-alkyl optionally substituted with one or two hydroxyl groups, C2-6-alkenyl optionally substituted with one or two hydroxyl groups, halo-C1-6-alkyl, cyclo-C3-6-alkyl, C1-6-alkoxy, halo-C1-6-alkoxy, C1-6-alkanoyl, R′OOC—C1-6-alkyl-, R′OOC—C1-6-alkoxy-, —NO2, —CN, NC—C1-6-alkyl-, halogen, R″R′N—C1-6-alkyl-, R″R′N—C1-6-alkoxy-, R″-C(O)—NR′—C1-6-alkyl-, R″R′N—C(O)—C1-6-alkyl-, R″-C(O)—NR′—C1-6-alkoxy-, R″R′N—C(O)—C1-6-alkoxy, —NR′R″, —NR′—C(O)—R″, —C(O)—NR″R′, C1-6-alkoxy-C1-6-alkyl- and C1-6-alkoxyC1-6-alkoxy-;
Ar2 is a ring or a fused ring system, in which the ring or the ring system is unsaturated or saturated, includes 5-12 atoms of which 0-4 are heteroatoms selected from N, O, and S, and is optionally substituted with one or more groups selected from C1-6-alkyl optionally substituted with one or more hydroxyl groups, C2-6-alkenyl optionally substituted with one or two hydroxyl groups, C1-6-alkanoyl, C1-6-alkoxy, C1-6-alkoxy-C1-6-alkyl- and halogen;
RB is a 3-membered hydrocarbon ring or a 4-, 5-, or 6-membered saturated or unsaturated ring with 0 to 3 heteroatoms independently selected from N, O and S and optionally substituted with one or more groups selected from C1-6-alkyl optionally substituted with one or two hydroxyl groups, C2-6-alkenyl optionally substituted with one or two hydroxyl groups, halo-C1-6-alkyl, cyklo-C3-6-alkyl, C1-6-alkoxy, C1-6-alkanoyl, R′OOC—C1-6-alkyl-, R′OOC—C1-6-alkoxy-, R″R′N—C1-6-alkyl-, R″R′N—C1-6-alkoxy-, —NR′R″, pyrrolidyl and halogen;
alternatively RB is selected from H, C1-6-alkyl optionally substituted with one or two hydroxyl groups, C2-6-alkenyl optionally substituted with one or two hydroxyl groups, halo-C1-6-alkyl, halogen, halo-C1-6-alkoxy, —NR′R″, C1-6-alkoxy and —CN;
R′ and R″ are independently selected from H, C1-6-alkyl optionally substituted with one or more hydroxyl groups, C2-6-alkenyl optionally substituted with one or two hydroxyl groups, halo-C1-6-alkyl, C1-6-alkanoyl and C1-6-alkoxyC1-6-alkyl;
provided that
(i) the sulphonamide derivative is not a compound of formula (I) where (a) X is methoxy-substituted phenyl and Ar2 is pentafluorophenyl, or (b) R1 is hydrogen and Ar1 is substituted phenyl; and
(ii) the sulphonamide derivative is not a compound of formula (I′), where L is —CH2— and Ar1 is phenyl.
Further the invention relates to derivatives of formula (I) and (I′) for use as inhibitors for collagen receptor integrins specifically α2β1 integrin inhibitors.
The invention also relates to derivatives of formula (I) and (I′) and physiologically acceptable salts thereof for use as a medicament.
Further the invention relates to the use of a derivative of formula (I) and (I′) for preparing a pharmaceutical composition for treating disorders relating to thrombosis, inflammation, cancer and vascular diseases.
The present invention also relates to a pharmaceutical composition comprising an effective amount of a derivative of formula (I) and (I′) or a physiologically acceptable salt thereof and one or more suitable adjuvant.
Further the invention relates to a process for preparing a sulphonamide derivative according to the invention, comprising
where R1, R2, R2′, R3, X, L, and Ar1 are as defined in claim 1, with a compound of formula (IV)
RB—Ar2—SO2-G (IV)
where RB and Ar2 is as defined in claim 1 and G is a leaving group, preferably a halogen;
reacting a compound of formula (V)
where R1, R2, R3, X, RB, and Ar2 are as defined in claim 1, with a compound of formula (VI)
G-C(O)NR2-L-Ar1 (VI)
where R2′, L and Ar1 are as defined in claim 1 and G is a leaving group, preferably a halogen; or
reacting a compound of formula (VII)
where Ar2, R1, R2, R2′, R3, X, L and Ar1 are as defined in claim 1 and G is a leaving group, preferably a halogen, with a compound of formula (VIII)
RB-M (VIII)
where RB is as defined above and M is a leaving group such as a metal.
The present invention relates to sulphonamide derivatives, with a urea moiety, having the general formula (I) or (I′). In the sulphonamide derivative of the present invention the sulphonamide moiety and the urea moiety is separated with a central aromatic ring as defined in claim 1. The sulphonamide moiety is either attached to the central aryl via the nitrogen atom (formula I) or sulphur atom (formula I′).
The central aromatic ring is preferably phenyl, pyrrolyl, furanyl, thiophenyl, pyridinyl or pyrimidinyl. More preferably the sulphonamide derivatives according to the current invention have the general structure Ia or Ia′
in which x′ is selected from —CH═CH—, —CH═N— and NR′.
In the formulas I and I′ as well as in Ia and Ia′ Ar1 is preferably an optionally substituted phenyl, Ar2 is preferably thiophenyl, pyrazolyl or phenyl and further R1 is preferably H, CH3, hydroxyethyl or hydroxypropyl. A sulphonamide derivative where R1 is CH3, X is —CH═CH—, R2 and R2′ are both H, L is absent and Ar1 is phenyl is also preferred.
In a preferred embodiment of the present invention the sulphonamide derivative is selected from the group consisting of:
Other typical sulphonamide derivatives of the present invention are presented in table 1.
The term “alkyl” used herein refers to a linear chain alkyl, such as methyl, ethyl, propyl and butyl groups, or branched alkyl group, such as isopropyl and isobutyl groups. Alkyl groups of the current invention typically have from 1 to 6 carbon atoms and preferably 1 to 3 carbon atoms.
The term “alkenyl” used herein refers to a linear or branched hydrocarbon group having at least one carbon-carbon double bond. Alkenyl groups of the current invention typically have from 2 to 6 carbon atoms and preferable 2 to 4 carbon atoms.
The term “alkanoyl” refers to branched or straight chain alkylcarbonyl groups, i.e. alkyl groups with a C═O group, having typically from 1 to 6 carbon atoms, preferably 1 to 3 carbon atoms.
The term “alkoxy” refers to branched or straight chain alkyloxy groups (—O-alkyl) having typically from 1 to 6 carbon atoms, preferably 1 to 3 carbon atoms, in the alkyl moiety.
The term “halo” or “halogen” refers to the non-metal elements of group 17 (IUPAC Style) and is selected from F, Cl, Br and I.
The term “saturated ring” refers to a cyclic hydrocarbon ring or a heterocyclyl with only divalent carbon atoms, i.e. a CH2 or a substituted CH2 group, in the ring.
The term “unsaturated ring” refers to aromatic rings or rings with at least one double bond, e.g. a carbon-carbon double bond or a carbon-nitrogen double bond.
The term “fused ring system” refers to a moiety where two or more hydrocarbon rings or heterocycles are fused together, e.g.
The term “linker” refers to a hydrocarbon chain linking two parts of a molecule together, the hydrocarbon chain typically contain 1 to 6 carbon atoms and can be linear, such as —CH2— and —CH2CH2—, or branched such as —CH2CH(CH3)CH2— and CH2CH(CH2CH3)CH2CH2—.
The term “cycloalkyl” refers to a cyclic hydrocarbon group having typically from 3 to 6 carbon atom, e.g. cyclopropyl and cyclobutyl.
Typical physiologically acceptable salts are e.g. acid addition salts (e.g. HCl, HBr, mesylate, etc.) and alkalimetal and alkaline earth metal salts (Na, K, Ca, Mg, etc.) conventionally used in the pharmaceutical field. Other suitable salts are e.g. ammonium, glucamine, amino acid etc. salts.
The pharmaceutical compositions can contain one or more of the sulphonamides of the invention. The administration can be parenteral, subcutaneous, intravenous, intraarticular, intrathecal, intramuscular, intraperitoneal or intradermal injections, or intravenous infusion, or by transdermal, rectal, buccal, oromucosal, nasal, ocular routes or via inhalation or via implant. Alternatively or concurrently, administration can be by the oral route. The required dosage will depend upon the severity of the condition of the patient, for example, and such criteria as the patient's weight, sex, age, and medical history. The dose can also vary depending upon whether it is to be administered in a veterinary setting to an animal or to a human patient.
For the purposes of parenteral administration, compositions containing the sulphonamides of the invention are preferably dissolved in sterile water for injection and the pH preferably adjusted to about 6 to 8 and the solution is preferably adjusted to be isotonic. If the sulphonamide is to be provided in a lyophilized form, lactose or mannitol can be added as a bulking agent and, if necessary, buffers, salts, cryoprotectants and stabilizers can also be added to the composition to facilitate the lyophilization process, the solution is then filtered, introduced into vials and lyophilized.
Useful excipients for the compositions of the invention for parenteral administration also include sterile aqueous and non-aqueous solvents. The compounds of the invention may also be administered parenterally by using suspensions and emulsions as pharmaceutical forms. Examples of useful non-aqueous solvents include propylene glycol, polyethylene glycol, vegetable oil, fish oil, and injectable organic esters. Examples of aqueous carriers include water, water-alcohol solutions, emulsions or suspensions, including saline and buffered medical parenteral vehicles including sodium chloride solution, Ringer's dextrose solution, dextrose plus sodium chloride solution, Ringer's solution containing lactose, or fixed oils. Examples of solubilizers and co-solvents to improve the aqueous properties of the active compounds to form aqueous solutions to form parenteral pharmaceutical dosage forms are propylene glycol, polyethylene glycols and cyclodextrins. Examples of intravenous infusion vehicles include fluid and nutrient replenishers, electrolyte replenishers, such as those based upon Ringer's dextrose and the like.
Injectable preparations, such as solutions, suspensions or emulsions, may be formulated according to known art, using suitable dispersing or wetting agents and suspending agents, as needed. When the active compounds are in water-soluble form, for example, in the form of water soluble salts, the sterile injectable preparation may employ a non-toxic parenterally acceptable diluent or solvent as, for example, water for injection (USP). Among the other acceptable vehicles and solvents that may be employed are 5% dextrose solution, Ringer's solution and isotonic sodium chloride solution (as described in the Ph. Eur/USP). When the active compounds are in a non-water soluble form, sterile, appropriate lipophilic solvents or vehicles, such as fatty oil, for example, sesame oil, or synthetic fatty acid esters, for example, ethyl oleate or triglycerides, are used. Alternatively, aqueous injection suspensions which contain substances which increase the viscosity, for example, sodium carboxymethyl cellulose, sorbitol, and/or dextran, and optionally also contain stabilizers may be used.
Pharmaceutical preparations for oral (but systemic) administration can be obtained by combining the active compounds with solid excipients, optionally granulating a resulting mixture and processing the mixture or granules or solid mixture without granulating, after adding suitable auxiliaries, if desired or necessary, to give tablets or capsules after filling into hard capsules.
Suitable excipients are, in particular, fillers such as sugars, for example lactose or sucrose, mannitol or sorbitol, cellulose and/or starch preparations and/or calcium phosphates, for example tricalcium phosphate or calcium hydrogen phosphate, as well as binders, such as starches and their derivatives, pastes, using, for example, maize starch, wheat starch, rice starch, or potato starch, gelatine, tragacanth, methyl cellulose, hydroxypropylmethyl cellulose, sodium carboxymethyl cellulose, and/or polyvinyl pyrrolidone, derivatives, and/or, if desired, disintegrating agents, such as the above-mentioned starches, and also carboxymethyl-starch, cross-linked polyvinyl pyrrolidone, agar or alginic acid or a salt thereof, such as sodium alginate. Auxiliaries are, above all, flow-regulating agents and lubricants, for example, silica, talc, stearic acid or salts thereof, such as magnesium stearate or calcium stearate, with suitable coating, which if desired, are resistant to gastric juices and for this purpose, inter alia concentrated sugar solutions, which optionally contain gum arabic, talc, polyvinyl pyrrolidone, polyethylene glycol and/or titanium dioxide, lacquer solutions and suitable organic solvents or solvent mixtures, but also film coating using e.g. cellulose derivatives, polyethylene glycols and/or PVP derivatives may be used. In order to produce coatings resistant to gastric juices, solutions of suitable cellulose preparations such as acetyl cellulose phthalate or hydroxypropylmethyl cellulose phthalate, are used for coating. Dyestuffs or pigments may be added to the tablets or dragee coatings or to coatings for example, for identification or in order to characterize different combinations of active compound doses.
Solid dosage forms for oral administration include capsules, tablets, pills, troches, lozenges, powders and granules. In such solid dosage forms, the active compound may be admixed with at least one inert diluent such as sucrose, lactose or starch. Such dosage forms may also comprise, as is normal practice, pharmaceutical adjuvant substances, e.g., stearate lubricating agents or flavouring agents. Solid oral preparations can also be prepared with enteric or other coatings which modulate release of the active ingredients.
Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups and elixirs containing inert non-toxic diluents commonly used in the art, such as water and alcohol. Such compositions may also comprise adjuvants, such as wetting agents, buffers, emulsifying, suspending, sweetening and flavouring agents.
The compositions of the invention may also be administered by means of pumps, or in sustained-release form. The compounds of the invention may also be delivered to specific organs in high concentration by means of suitably inserted catheters, or by providing such molecules as a part of a chimeric molecule (or complex) which is designed to target specific organs.
Administration in a sustained-release form is more convenient for the patient when repeated injections for prolonged periods of time are indicated so as to maximize the comfort of the patient. Controlled release preparation can be achieved by the use of polymers to complex or adsorb the cornpounds of the invention. Controlled delivery can be achieved by selecting appropriate macromolecules (for example, polyesters, polyamino acids, polyvinyl pyrrolidone, ethylenevinylacetate, methylcellulose, carboxymethylcelluloase protamine zinc and protamine sulfate) as well as the method of incorporation in order to control release. Another possible method to control the duration of action by controlled release preparations is to incorporate the desired compounds into particles of a polymeric material such as polyesters, polyamino acids, hydrogels, poly (lactic acid) or ethylene vinylacetate copolymers. Alternatively, instead of incorporating the sulphonamide into these polymeric particles, the sulphonamide can be entrapped into microparticles, prepared, for example, by coacervation techniques or by interfacial polymerization, for example, hydroxymethylcellulose or gelatin-microcapsules and poly (methylmethacrylate) microcapsules, respectively, or in colloidal drug delivery systems, for example liposomes, albumin microspheres, microemulsions, nanoparticles, and nanocapsules or in macroemulsions. The above-mentioned technique may be applied to both parenteral and oral administration of the pharmaceutical formulation.
The pharmaceutical compositions of the present invention can be manufactured in a manner which is in itself known, for example, by means of conventional mixing, granulating, dragee-making, dissolving, lyophilizing or similar processes.
The compounds of the invention are potent collagen receptor inhibitors and useful for inhibiting or preventing the adhesion of cells on collagen or the migration and invasion of cells through collagen containing matrices, in vivo or in vitro. The now described compounds inhibit the migration of malignant cells and are thus for treating diseases such as cancers, including prostate, and melanoma, especially where α2β1 integrin dependent cell adhesion/invasion/migration may contribute to the malignant mechanism.
The compounds of the invention also inhibit inflammatory responses that require α2β1 integrin. The now described compounds can inhibit pathological inflammatory processes and can thus be used for treating diseases like inflammatory bowel disease, psoriasis, arthritis, multiple sclerosis, asthma, and allergy.
The compounds of the invention also inhibit adhesion of platelets to collagen and collagen-induced platelet aggregation. Thus, the compounds of the invention are useful for treating patients in need of preventative or ameliorative treatment for thromboembolic conditions i.e. diseases that are characterized by a need to prevent adhesion of platelets to collagen and collagen-induced platelet aggregation, for example treatment and prevention of stroke, myocardial infraction unstable angina pectoris diabetic rethinopathy or retinal vein occlusion.
Chinese Hamster Ovary (CHO) cell clone expressing wild type α2 integrin was used in cell adhesion assay. Cells were suspended in serum free medium containing 0.1 mg/mL cycloheximide (Sigma) and the compounds were preincubated with the cells prior to transfer to the wells. Cells (150000/well) were allowed to attach on collagen type I coated wells (in the presence and absence of inhibitor compounds) for 2 h at +37° C. and after that non-adherent cells were removed. Fresh serum free medium was added and the living cells were detected using a cell viability kit (Roche) according to the manufacturer's protocol.
The following examples illustrate the invention but are not intended to limitate the scope of the invention (Table 1).
The ability to interact with extracellular matrix basement membranes is essential for the malignant cancer cell phenotype and cancer spread. α2β1 levels are known to be upregulated in tumorigenic cells. The overexpression regulates cell adhesion and migration to and invasion through the extracellular matrix. By blocking the interaction between extracellular matrix components like collagen and α2β1 it is possible to block cancer cell migration and invasion in vitro. Prostate cancer cells (PC-3) expressing α2β1 endogenously were used to test the in vitro anticancer potential of the inhibitors of the present invention.
Invasion of PC-3 cells (CRL-1435, ATCC) through Matrigel was studied using BD Biocoat invasion inserts (BD Biosciences). Inserts were stored at −20° C. Before the experiments inserts were allowed to adjust to the room temperature. 500 μl of serum free media (Ham's F12K medium, 2 mM Lglutamine, 1.5 g/l sodium bicarbonate) was added into the inserts and allowed to rehydrate at 37° C. in cell incubator for two hours. The remaining media was aspirated. PC-3 cells were detached, pelleted and suspended into serum free media (50 000 cells/500 μL). 300 μL of cell suspension was added into the insert in the absence (control) or presence of the inhibitor according to the present invention. Inserts were placed on the 24-well plates; each well containing 700 μL of cell culture media with 3% of fetal bovine serum as chemoattractant. Cells were allowed to invade for 72 hours at 37° C. in cell incubator. Inserts were washed with 700 μL PBS, and fixed with 4% paraformaldehyde for 10 minutes. Paraformaldehyde was aspirated and cells were washed with 700 μL of PBS and inserts were stained by incubation with hematoxylin for 1 minute. The stain was removed by washing the inserts with 700 μL of PBS. Inserts were allowed to dry. Fixed invaded cells were calculated under the microscope. Invasion % was calculated as a comparison to the control.
Cell invasion assay is used as an in vitro cancer metastatis model. The sulfonamide molecules have been shown to inhibit tumor cell invasion in vitro (see table 2). Some structures inhibit invasion even with submicromolar concentrations.
Cellix system was used to demonstrate the possible antithrombotic effects of α2β1 modulators in flow conditions. Cellix microfluidic platform models human blood vessels providing a dynamic set-up mimicking physiological conditions to test new therapeutic agents (Cellix Ltd). The platform was used to measure the platelet adhesion to collagen coated capillary under flow. An anti-coaculated whole blood sample was run through a collagen coated capillary under a constant shear and the size of thrombi on capillary wall was analyzed with analysis program (DucoCell, Cellix Ltd). If the average thrombi area was decreased when compared to the control sample the compound was suggested to have antithrombotic activity.
The capillary were coated with Horm collagen 20 μg/mL (Nycomed) and incubated for 24 h in +4° C. Background was blocked with 1% BSA (bovine serum albumin, Sigma) treatment for 30 min in room temperature. Blood was collected from a donor into blood collection tubes containing 40 μM PPACK (Dphenylalanyl-L-prolyl-L-arginine chloromethyl-ketone, Calbiochem) as anticoagulant. Blood was treated either with inhibitory compounds or vehicle controls. Samples were kept at room temperature for 5 minutes. The samples were run through the capillary with the constant shear rate (90 dynes/cm2, Mirus 1.0 Nanopump, Cellix Ltd) for 5 min and capillary was washed with JNL buffer (6 mM Dextrose, 0.13 M NaCl, 9 mM Na Bicarb, 10 mM Na Citrate, 10 mM Tris base, 3 mM KCl, 0.81 mM KH2PO4, 0.9 mM MgCl2; pH was adjusted to 7.35 with 19 mM Citrate acid anhydrous, 37 mM Sodium citrate, 67 mM Dextrose) with the constant shear rate (90 dynes/cm2) for 2 min. After that the average thrombi area on the capillary wall was analyzed with DucoCell analysis program (Cellix Ltd). See table 3.
A platelet function analyzer PFA-100 was used to demonstrate the possible antithrombotic effects of α2β1 modulators. The PFA-100 is a high shear-inducing device that simulates primary haemostasis after injury of a small vessel. The system comprises a test-cartridge containing a biologically active membrane coated with collagen and ADP. An anticoaculated whole blood sample was run through a capillary under a constant vacuum. The platelet agonist (ADP) on the membrane and the high shear rate resulted in activation of platelet aggregation, leading to occlusion of the aperture with a stable platelet plug. The time required to obtain full occlusion of the aperture was designated as the closure time. Each compound was added to the whole blood sample and the closure time was measured with PFA-100. If the closure time was increased when compared to the control sample the compound was suggested to have antithrombotic activity.
Blood was collected from a donor via venipuncture into evacuated blood collection Lithium heparin tubes (VenoJect, Terumo). Blood was aliquoted into 1.5 mL tubes and treated with either inhibitory compounds or controls (DMSO). Samples were kept at room temperature with rotation for 10 minutes and after that the closure time (Ct) of the blood was measured.
Results from the experiments show that the derivatives according to the invention increase the closure time (Ct) of the whole blood in PFA-100 analysis (see
Interleukin-6 was measured to demonstrate the anti-inflammatory potential of the α2β1 inhibitors.
The inflammatory cells are shown to be dependent on α2β1 function during inflammatory process. The anti-inflammatory potential of the α2β1 modulators was studied by measuring the effect on the release of cytokines like IL-6 as an example from inflammatory cells. Lipopolysaccharide (LPS) was used to induce the production of IL-6. Anticoaculated whole blood sample was incubated with or without the integrin α2β1 inhibitor before LPS was added to induce the cytokine release. The amount of cytokine IL-6 released from peripheral blood leukocytes was measured from plasma samples 2 hours after the induction.
Blood was collected from the donors via venipuncture into evacuated Lithium heparin tubes (VenoJect, Terumo). Blood was treated with either α2β1 integrin inhibitors or vehicle control (DMSO). Samples were kept at room temperature for 5 minutes and lipopolysaccharide (LPS, 0.25 ng/mL) was added to induce cytokine release from peripheral blood lymphocytes. Samples were inculabted for 2 h in 37° C., the plasma was isolated and the amount of IL-6 was determined with Human IL-6 Quantikine ELISA Kit (R&D Systems).
Results from the experiments show that the derivatives according to the invention statistically significantly decrease the IL-6 release after incuction with LPS (see
1H NMR spectra were recorded at ambient temperature using a Varian Unity Inova (400 MHz) spectrometer with a triple resonance 5 mm probe for example compounds, and either a Bruker Avance DRX (400 MHz) spectrometer or a Bruker Avance DPX (300 MHz) spectrometer for intermediate compounds. Chemical shifts are expressed in ppm relative to tetramethylsilane. The following abbreviations have been used: br=broad signal, s=singlet. d=doublet, dd=double doublet, dt=double triplet, t=triplet, q=quartet, m=multiplet.
High pressure liquid chromatography-Mass Spectrometry (LCMS) experiments to determine retention times (Rt) and associated mass ions were performed using one of the following methods:
Method A: Experiment performed on a Waters Platform LC quadrupole mass spectrometer linked to a Hewlett Packard HP1100 LC system with diode array detector and 100 position autosampler. The spectrometer has an electrospray source operating in positive and negative ion mode. Additional detection is achieved using a Sedex 85 evaporative light scattering detector. Samples are run through a LC column-Phenomenex Luna 3 micron C18(2) 30×4.6 mm and a 2 ml/min flow rate. The initial solvent system was 95% water containing 0.1% formic acid (solvent A) and 5% acetonitrile containing 0.1% formic acid (solvent B) for the first half minute followed by a gradient up to 5% solvent A and 95% solvent B over the next 4 minutes. The final solvent system was held constant for a further minute.
Method B: Experiment performed on a Waters ZMD quadrupole mass spectrometer linked to a Waters 1525 LC system with Waters 996 diode array detector. Sample injection is done by a Waters 2700 autosampler. The spectrometer has an electrospray source operating in positive and negative ion mode. Additional detection is achieved using a Sedex 85 evaporative light scattering detector. Samples are run through a LC column-Luna 3 micron C18(2) 30×4.6 mm and a 2 ml/min flow rate. The initial solvent system was 95% solvent A and 5% solvent B for the first half minute, followed by a gradient up to 5% solvent A and 95% solvent B over the next 4 minutes. The final solvent system was held constant for a further minute.
Method C: Experiment performed on a Waters Micromass ZQ2000 quadrupole mass spectrometer linked to a Hewlett Packard HP 1100 LC system with a DAD UV detector. Sample injection is done by a CTC HTS PAL autosampler. The spectrometer has an electrospray source operating in positive and negative ion mode. Additional detection is achieved using a Sedex 85 evaporative light scattering detector. Samples are run through a LC columnHiggins Clipeus 5 micron C18 100×3.0 mm and a 1 ml/min flow rate. The initial solvent system was 95% solvent A and 5% solvent B for the first minute, followed by a gradient up to 5% solvent A and 95% solvent B over the next 14 minutes. The final solvent system was held constant for a 5 further minutes.
Microwave experiments were carried out using a Biotage Initiator™, which uses a single-mode resonator and dynamic field tuning, both of which give reproducibility and control. Temperatures from 40-250° C. can be achieved, and pressures of up to 20 bars can be reached. Three types of vial are available for this processor, 0.5-2.0 ml, 2.0-5.0 ml and 5.0-20 ml.
Preparative HPLC purification was carried out using a C18-reversephase column (100×22.5 mm i.d. Genesis column with 7 μm particle size, UV detection at 230 or 254 nm, flow 5-15 ml/min), eluting with gradients from 100-0 to 0-100% water/acetonitrile containing 0.1% formic acid, with a flow rate of 18 ml per minute. Fractions containing the required product (identified by LCMS analysis) were pooled, the organic fraction removed by evaporation, and the remaining aqueous fraction lyophilised, to give the final product.
Compounds which required column chromatography were purified manually or fully automatically using either a Biotage SP1™ Flash Purification system with Touch Logic Control™ or a Combiflash Companion® with prepacked silica gel Isolute® SPE cartridge, Biotage SNAP cartridge or Redisep® Rf cartridge respectively.
Abbreviations:
DCM—Dichloromethane
DMF—N, N-Dimethylformamide
THF—Tetrahydrofuran
DMAP—4-Dimethylaminopyridine
TFA—Trifluoroacetic acid
Boc—tert-Butoxycarbonyl
IMS—Industrial methylated spirits
NMP—N-methylpyrrolidinone
(3-Aminophenyl)carbamic acid tert-butyl ester (1.0 g) was dissolved in ethyl acetate (30 ml) and treated with aqueous formaldehyde (37% wt, 443 μl) and palladium on carbon (10%, 350 mg). The reaction mixture was hydrogenated under a balloon of hydrogen overnight at atmospheric pressure. The catalyst was removed by filtration through Celite under nitrogen and the volatiles were removed by evaporation. The residue was purified by chromatography using the Biotage system on a 10 g silica cartridge eluting with a mixture of ethyl acetate and cyclohexane (1:19 increasing to 1:1) to give (3-methylaminophenyl)carbamic acid tert-butyl ester (500 mg).
LCMS (Method A) Rt 2.45 (M+H+) 223
1H NMR (300 MHz) (CDCl3) δ 7.2 (t, 1H) 6.8 (br s, 1H) 6.5 (dd, 1H) 6.4 (br s, 1H), 6.3 (dd, 1H) 3.7 (br s, 1H) 2.8 (s, 3H) 1.5 (s, 9H)
To a solution of 4′-fluorobiphenyl-3-sulphonyl chloride (245 mg) in pyridine (5 ml) was added a solution of (3-methylaminophenyl)carbamic acid tert-butyl ester (Intermediate 1, 200 mg) in pyridine (2 ml), and the resultant mixture was stirred at room temperature for 3 hours. The pyridine was removed by evaporation under reduced pressure and the residue was partitioned between water and ethyl acetate. The organic layer was dried (Na2SO4), filtered and the volatiles were removed by evaporation. The residue was purified by HPLC eluting with a mixture of water and acetonitrile (each containing 0.1% formic acid) from 20 to 98% acetonitrile over 25 minutes to give {3-[N-(4′-Fluorobiphenyl-3-sulphonyl)-N-methylamino]-phenyl}carbamic acid tert-butyl ester as a white solid (220 mg).
LCMS (Method A) Rt 4.26 (M−H) 455
1H NMR (300 MHz) (CDCl3) δ 7.75 (dd, 1H) 7.7 (br s, 1H) 7.6-7.5 (m, 2H) 7.5-7.4 (m, 2H) 7.35 (d, 1H) 7.2 (m, 2H) 7.15 (t, 2H) 6.75 (dd, 1H) 6.5 (br s, 1H) 3.2 (s, 3H) 1.5 (s, 9H)
By proceeding in a similar manner the following intermediates were prepared from the appropriate starting materials. The reaction may also be performed in DCM as a solvent in the presence of a base such as pyridine.
From 4′-fluorobiphenyl-3-sulphonyl chloride and (4-aminophenyl)carbamic acid tert-butyl ester
LCMS (Method A) Rt 4.09 (M−H) 441
1H NMR (400 MHz) (CDCl3) δ 7.8 (s, 1H) 7.7 (m, 2H) 7.5-7.4 (m, 3H) 7.2 (m, 3H) 7.0 (d, 2H) 6.6 (d, 2H) 6.4 (br s, 1H) 1.5 (s, 9H)
From 4′-fluorobiphenyl-3-sulphonyl chloride and 4-nitroaniline LCMS (Method A) Rt 3.88 (M−H) 371
1H NMR (300 MHz) (CDCl3) δ 8.2 (d, 2H) 8.1 (s, 1H) 7.8 (dd, 1H) 7.7 (dd, 1H) 7.6 (t, 1H) 7.5 (m, 2H) 7.2 (m, 3H) 7.1 (t, 2H)
From 4′-fluorobiphenyl-3-sulphonyl chloride and 2-methoxy-5-nitroaniline
LCMS (Method A) Rt 3.86 (M−H) 401
From 5-bromopyridine-3-sulphonyl chloride and 1-(4-methylaminophenyl)-3-phenyl urea (Intermediate 25)
LCMS (Method A) Rt 3.47 (M+H+) 338
From 3-bromophenylsulphonyl chloride and 1-(4-methylaminophenyl)-3-phenyl urea (Intermediate 25)
LCMS (Method A) Rt. 3.87 (M+H+) 462
1H NMR (300 MHz) (DMSO-d6) δ 8.8 (s, 1H) 8.7 (s, 1H) 7.9 (dd, 1H) 7.6 (m, 3H) 7.4 (m, 4H) 7.3 (t, 2H) 7.0 (m, 3H) 3.1 (s, 3H)
From 1-(4-fluorophenyl)-1H-pyrazole-4-sulphonyl chloride (Intermediate 87) and 1-(4-aminophenyl)-3-phenyl urea (Intermediate 23)
LCMS (Method C) Rt 10.56 (M+H+) 452
1H NMR (400 MHz) (DMSO-d6) δ 10.0 (br s, 1H) 8.9 (s, 1H) 8.6 (br s, 2H) 7.9 (m, 3H) 7.4 (m, 6H) 7.3 (t, 2H) 7.1 (d, 2H) 6.9 (t, 1H)
From 5-(4,4,4-trifluoro-1,3-dioxobutyl)thiophene-2-sulphonyl chloride and 1-(4-methyl-aminophenyl)-3-phenylurea (Intermediate 25)
Intermediate used without purification or characterisation
From 4′-fluorobiphenyl-3-sulphonyl chloride and N-methyl-4-nitroaniline using pyridine in DCM.
1H NMR (400 MHz) (CDCl3) δ 8.2 (d, 2H) 7.75 (dt, 1H) 7.70 (t, 1H) 7.5 (t, 1H) 7.45 (m, 3H) 7.35 (d, 2H) 7.15 (t, 2H) 3.3 (s, 3H)
From 5-(1-methyl-3-trifluoromethyl-1H-pyrazol-5-yl)thiophene-2-sulphonyl chloride and 2-methoxy-4-nitroaniline using pyridine in DCM.
1H NMR (400 MHz) (DMSO-d6) δ 10.7 (br s, 1H) 7.9 (dd, 1H) 7.8 (d, 1H) 7.75 (d, 1H) 7.65 (d, 1H) 7.55 (d, 1H) 7.2 (s, 1H) 4.0 (s, 3H) 3.8 (s, 3H)
From 4′-fluorobiphenyl-3-sulphonyl chloride and 2-methyl-4-nitroaniline using pyridine in DCM.
1H NMR (300 MHz) (CDCl3) δ 8.2 (d, 1H) 8.0 (t, 1H) 7.9 (dd, 1H) 7.7 (m, 2H) 7.6 (t, 1H) 7.5 (m, 2H) 7.3 (d, 1H) 7.2 (t, 2H) 6.5 (br s, 1H) 2.2 (s, 3H)
From 4′-fluorobiphenyl-3-sulphonyl chloride and 2-methoxy-4-nitroaniline using pyridine in DCM.
1H NMR (400 MHz) (CDCl3) δ 8.0 (t, 1H) 7.85 (m, 2H) 7.75 (dt, 1H) 7.66 (s, 1H) 7.64 (d, 1H) 7.55 (t, 1H) 7.48 (m, 3H) 7.15 (t, 2H) 3.8 (s, 3H)
From 4-nitrophenylsulphonyl chloride and 3-bromoaniline using pyridine in DCM.
1H NMR (300 MHz) (DMSO-d6) δ 10.90 (br s, 1H), 8.40 (d, 2H) 8.02 (d, 2H) 7.31-7.21 (m, 3H) 7.13 (dt, 1H).
From 4-bromo-5-chlorothiophene-2-sulphonyl chloride and 1-(4-methylaminophenyl)-3-phenyl urea, (Intermediate 25)
1H NMR (300 MHz) (DMSO-d6) δ 8.8 (br s, 1H), 8.7 (br s, 1H) 7.65 (s, 1H) 7.5 (dd, 4H) 7.3 (t, 2H) 7.15 (d, 2H) 7.0 (t, 1H) 3.2 (s, 3H)
From 1-H-imidazole-4-sulphonyl chloride and 1-(4-methylaminophenyl)-3-phenyl urea (Intermediate 25) using pyridine in DCM.
LCMS (Method A) Rt 2.90 (M+H+) 372
From 1H-pyrazole-4-sulphonyl chloride and 1-(4-methylaminophenyl)-3-phenyl urea (Intermediate 25) using pyridine in DCM.
LCMS (Method C) Rt 8.55 (M+H+) 372
1H NMR (400 MHz) (DMSO-d6) 8.85 (s, 1H), 8.75 (s, 1H) 7.9 (s, 2H) 7.4 (m, 4H) 7.25 (t, 2H) 7.0 (d, 2H) 6.95 (t, 1H) 3.1 (s, 3H).
To a stirred solution of N-[(2,2-dimethyl-1,3-dioxolan-4-yl)methyl]-4-nitroaniline (Intermediate 19, 300 mg) in DMF (9 ml) was added portionwise NaH (60% in mineral oil, 160 mg). The mixture was stirred for 15 minutes and then a solution of 4′-fluorobiphenyl-3-sulphonyl chloride (390 mg) in DMF (1 ml) was added. Stirring was continued for a further 4 hours then the reaction mixture was quenched with water and extracted with ethyl acetate. The organic layer was washed with water, dried (Na2SO4), filtered and the volatiles were removed by evaporation. The residue was purified by chromatography using the Companion on a 20 g silica cartridge, eluting with a mixture of ethyl acetate and cyclohexane (1:9 increasing to 1:4), to give 4′-fluorobiphenyl-3-sulphonic acid N-[(2,2-dimethyl-1,3-dioxolan-4-yl)methyl]-N-(4-nitrophenyl)amide as a yellow oil (320 mg).
1H NMR (300 MHz) (DMSO-d6) δ 8.2 (d, 2H) 7.8 (dt, 1H) 7.75 (t, 1H) 7.55 (t, 1H) 7.5 (m, 3H) 7.35 (d, 2H) 7.15 (m, 2H) 4.25 (m, 1H) 4.05 (dd, 1H) 3.8 (dd, 1H) 3.75 (dd, 2H) 1.25 (s, 6H)
A solution of (2,2-dimethyl-1,3-dioxolan-4-yl)methylamine (2.5 ml), 1-fluoro-4-nitrobenzene (1.41 g) and triethylamine (1.4 ml) in ethanol (25 ml) was heated under reflux for 18 hours. The reaction mixture was cooled and the volatiles were removed by evaporation. The residue was purified by chromatography using the Companion on a 50 g silica cartridge, eluting with a mixture of ethyl acetate and pentane (1:4) to give N-[(2,2-dimethyl-1,3-dioxolan-4-yl)methyl]-4-nitroaniline as a yellow solid (533 mg).
1H NMR (400 MHz) (CDCl3) δ 8.1 (d, 2H) 6.6 (d, 2H) 4.8 (br s, 1H) 4.4 (m, 1H) 4.1 (dd, 1H) 3.8 (dd, 1H) 3.4 (m, 1H) 3.3 (m, 1H) 1.5 (s, 3H) 1.4 (s, 3H)
N-Methyl-4-nitroaniline (1.5 g) was dissolved in THF (100 ml) and boc anhydride was slowly added followed by DMAP. The mixture was stirred at room temperature for 4 hours then the volatiles were removed by evaporation. The residue was partitioned between water and ethyl acetate and the organic layer was washed with aqueous HCl, dried (Na2SO4) and filtered. The volatiles were removed by evaporation to give N-methyl-N-(4-nitrophenyl)carbamic acid tert-butyl ester as a pale beige solid (2.29 g)
LCMS (Method A) Rt 3.84 (M+H++CH3CN) 294
1H NMR (400 MHz) (CDCl3) δ 8.2 (d, 2H) 7.5 (d, 2H) 3.3 (s, 3H) 1.5 (s, 9H)
{3-[N-(4′-Fluorobiphenyl-3-sulphonyl)-N-methylamino]phenyl}carbamic acid tert-butyl ester (Intermediate 2, 200 mg) was dissolved in DCM (6 ml) and treated with TFA (2 ml). The mixture was stirred at room temperature for 2 hours, then the volatiles were removed by evaporation. The residue was partitioned between saturated aqueous NaHCO3 and ethyl acetate and the organic layer was dried (Na2SO4), filtered and the volatiles were removed by evaporation to give 4′-fluorobiphenyl-3-sulphonic acid N-(3-aminophenyl)-N-methylamide (169 mg).
LCMS (Method B) Rt 3.72 (M+H+) 357
1H NMR (300 MHz) (CDCl3) δ 7.7 (m, 2H) 7.6-7.4 (m, 4H) 7.2 (m, 3H) 6.6 (m, 2H) 6.4 (dd, 1H) 3.7 (br s, 2H) 3.3 (s, 3H)
By proceeding in a similar manner the following intermediates were prepared from the appropriate starting materials:
From {4-[N-(4′-fluorobiphenyl-3-sulphonyl)amino]phenyl}carbamic acid tert-butyl ester (Intermediate 3)
LCMS (Method B) Rt 2.95 (M+H+) 343
1H NMR (300 MHz) (CDCl3) δ 7.8 (s, 1H) 7.7 (m, 2H) 7.5 (m, 3H) 7.1 (t, 2H) 6.8 (d, 2H) 6.6 (m, 4H) 6.1 (s, 1H)
From N-[4-(3-phenylureido)phenyl]carbamic acid tert-butyl ester (Intermediate 44)
LCMS (Method B) Rt 0.32 & 1.84 (M+H+) 228
1H NMR (300 MHz) (DMSO-d6) δ 8.5 (br s, 1H) 8.1 (br s, 1H) 7.4 (d, 2H) 7.3 (t, 2H) 7.1 (d, 2H) 6.9 (t, 1H) 6.5 (d, 2H) 4.8 (br s, 2H)
From N-[3-(3-phenylureido)phenyl]carbamic acid tert-butyl ester (Intermediate 37)
1H NMR (400 MHz) (DMSO-d6) δ 8.5 (s, 1H) 8.3 (s, 1H) 7.4 (d, 2H) 7.25 (t, 2H) 6.95 (t, 1H) 6.85 (t, 1H) 6.75 (t, 1H) 6.55 (dd, 1H) 6.2 (dd, 1H) 5.0 (br s, 2H)
From N-methyl-N-[4-(3-phenylureido)phenyl]carbamic acid tert-butyl ester (Intermediate 36)
LCMS (Method B) Rt 1.87 (M+H+) 242
1H NMR (400 MHz) (DMSO-d6) δ 8.9 (br s, 2H) 7.5 (m, 4H) 7.3 (t, 2H) 7.0 (d, 2H) 6.9 (t, 1H) 4.0 (br s, 1H) 2.8 (s, 3H)
From N-[4-(3-benzylureido)phenyl]carbamic acid tert-butyl ester (Intermediate 41)
LCMS (Method A) Rt 0.37 & 1.96 (M+H+) 242
To a suspension of 1-[4-(5-chloro-1,3-dioxo-1,3-dihydroisoindol-2-yl)-3-methylphenyl]-3-phenyl urea (Intermediate 56, 811 mg) in ethanol (10 ml) was added hydrazine hydrate (2 ml). The mixture was stirred and heated under reflux for 20 minutes, then the volatiles were removed by evaporation. The residue was redissolved in ethyl acetate (10 ml) and heated under reflux for further 20 minutes. The reaction mixture was cooled, filtered and the filtrate was concentrated. The residue was purified by HPLC eluting with a mixture of water and acetonitrile (each containing 0.1% formic acid) from 30 to 60% over 20 minutes to give 1-(4-amino-3-methylphenyl)-3-phenyl urea as a white powder.
LCMS (Method A) Rt 2.03 (M+H+) 242
4′-Fluorobiphenyl-3-sulphonic acid N-(4-nitrophenyl)amide (Intermediate 4, 200 mg) was dissolved in THF (4.0 ml) and triphenyl phosphine (280 mg) was added, followed by 3-benzyloxypropan-1-ol (170 μl). The mixture was then cooled to 5° C. and diethyl azodicarboxylate (168 μl) was slowly added. The reaction mixture was stirred overnight at room temperature. The volatiles were removed by evaporation and the residue was purified by chromatography on a 5 g silica cartridge eluting with a mixture of ethyl acetate and cyclohexane (1:4) to give 4′-fluorobiphenyl-3-sulphonic acid N-(3-benzyloxypropyl)-N-(4-nitrophenyl)amide as a yellow oil (236 mg).
1H NMR (300 MHz) (CDCl3) δ 8.4 (d, 2H) 7.8 (m, 2H) 7.5-7.4 (m, 4H) 7.4-7.2 (m, 7H) 7.1 (t, 2H) 4.4 (s, 2H) 3.8 (t, 2H) 3.5 (t, 2H) 1.8 (t, 2H)
By proceeding in a similar manner the following compounds were prepared from the appropriate starting materials:
From 4′-fluorobiphenyl-3-sulphonic acid (2-methoxy-4-nitrophenyl)amide (Intermediate 13) and 3-tert-butoxypropan-1-ol
1H NMR (400 MHz) (DMSO-d6) δ 8.0 (d, 1H) 7.9 (dd, 1H) 7.8 (t, 1H) 7.75 (d, 1H) 7.7 (m, 3H) 7.65 (d, 1H) 7.5 (d, 1H) 7.3 (t, 2H) 3.65 (t, 2H) 3.5 (s, 3H) 3.2 (t, 2H) 1.45 (m, 2H) 1.1 (s, 9H)
From 3-nitrophenol and 2-methoxyethanol
1H NMR (300 MHz) (DMSO-d6) δ 7.9 (dd, 1H) 7.8 (d, 1H) 7.4 (t, 1H) 7.3 (dd, 1H) 4.2 (t, 2H) 3.7 (t, 2H) 3.5 (s, 3H)
A solution of 4′-fluorobiphenyl-3-sulphonic acid (4-nitrophenyl)amide (Intermediate 4, 100 mg) in THF (3 ml) was added to an ice-cooled suspension of NaH (60% in mineral oil, 14 mg) in THF (2 ml). The mixture was stirred for 15 minutes at 0° C. and then treated with ethyl bromoacetate (83 μl). The resultant mixture was stirred at room temperature for 2.5 hours followed by heating at 50° C. overnight. The volatiles were removed by evaporation and the residue was diluted with water, acidified with aqueous HCl and extracted with ethyl acetate. The organic layer was dried (Na2SO4), filtered and the volatiles were removed by evaporation. The reaction was repeated using 4′-fluorobiphenyl-3-sulphonic acid (4-nitrophenyl)amide (Intermediate 4, 601 mg) in THF (15 ml), NaH (84 mg) in THF (30 ml) and ethyl bromoacetate (500 μl). The crude residues from both experiments were combined and purified by chromatography using the Biotage system on a 10 g silica cartridge eluting with a mixture of ethyl acetate and cyclohexane (1:9) to give [N-(4′-fluorobiphenyl-3-sulphonyl)-N-(4-nitrophenyl)amino]acetic acid ethyl ester (780 mg).
1H NMR (300 MHz) (CDCl3) δ 8.2 (d, 2H) 7.9 (s, 1H) 7.8 (dd, 1H) 7.7 (dd, 1H) 7.6-7.4 (m, 5H) 7.1 (t, 2H) 4.5 (s, 2H) 4.2 (q, 2H) 1.2 (t, 3H)
By proceeding in a similar manner the following compound was prepared from the appropriate starting materials:
From 4′-fluorobiphenyl-3-sulphonic acid (2-methyl-4-nitrophenyl)amide (Intermediate 12) and iodomethane
LCMS (Method B) Rt 3.98 (M−H) 401
A mixture of 5-bromothiophene-2-sulphonic acid N-methyl-N-[4-(3-phenylureido)phenyl]-amide (Example 55, 93 mg), 1-(tetrahydropyran-2-yl)-1H-pyrazole-5-boronic acid pinacol ester (48 mg), tetrakis(triphenylphosphine)palladium(0) (14 mg), Na2CO3 (2M, 1.3 ml) and DME (2 ml) was heated in the microwave at 150° C. for 20 minutes. The resultant mixture was partitioned between water and ethyl acetate. The organic layer was dried (MgSO4), filtered and the volatiles were removed by evaporation. The residue was purified by chromatography on a 5 g silica cartridge eluting with a mixture of ethyl acetate and DCM (1:49 increasing to 1:9) to give 5-[1-(tetrahydropyran-2-yl)-1H-pyrazol-5-yl]thiophene-2-sulphonic acid N-methyl-N-[4-(3-phenylureido)phenyl]-amide (62 mg).
LCMS (Method B) Rt 3.24 (M+H+) 376
By proceeding in a similar manner the following intermediates were prepared from the appropriate starting materials. The reaction may also be performed using different catalysts, bases and solvents.
From 4-fluorophenylboronic acid and 4-nitrophenylsulphonic acid (3-bromophenyl)amide (Intermediate 14) using [1,1-Bis(diphenylphosphino)ferrocene]dichloropalladium (II) and Cs2CO3, in a mixture of DME and IMS.
The compound was used without purification or characterisation.
From 5-bromothiophene-2-sulphonic acid N-methyl-N-[4-(3-phenylureido)phenyl]-amide (Example 55) and 1-boc-3,6-dihydro-2H-pyridine-4-boronic acid pinacol ester.
LCMS (Method B) Rt 4.26 (M+H+) 569
N-(4-Aminophenyl)-N-methylcarbamic acid tert-butyl ester (Intermediate 62, 1.80 g) was dissolved in THF (50 ml) and treated with NaOH (1M, 8.5 ml) and phenyl isocyanate (715 μl). The resultant mixture was stirred at room temperature for 2 hours and the volatiles were removed by evaporation. The residue was acidified to pH 5 and extracted with ethyl acetate. The organic layer was dried (Na2SO4), filtered and the volatiles were removed by evaporation. The residue was purified by chromatography using the Biotage system on a 50 g silica cartridge eluting with a mixture of ethyl acetate and cyclohexane (1:19 increasing to 2:3) to give N-methyl-N-[4-(3-phenylureido)phenyl]carbamic acid tert-butyl ester as a white solid (2.74 g).
LCMS (Method B) Rt 3.64 (M+H+) 340
1H NMR (400 MHz) (CDCl3) δ 7.4 (m, 4H) 7.2 (m, 1H) 7.1 (m, 4H) 6.9 (br s, 1H) 6.8 (br s, 1H) 3.2 (s, 3H) 1.5 (s, 9H)
By proceeding in a similar manner the following compounds were prepared from the appropriate starting materials:
From N-(3-aminophenyl)carbamic acid tert butyl ester and phenyl isocyanate
LCMS (Method B) Rt 3.65 (M+H+) 328
From 4′-fluorobiphenyl-3-sulphonic acid N-(3-tert-butoxypropyl)-N-(2-methoxy-4-amino-phenyl)amide (Intermediate 75) and phenyl isocyanate
1H NMR (400 MHz) (DMSO-d6) δ 8.8 (s, 1H) 8.7 (s, 1H) 7.95 (d, 1H) 7.7 (m, 5H) 7.45 (d, 2H) 7.3 (m, 5H) 7.05 (d, 1H) 7.0 (t, 1H) 6.9 (dd, 1H) 3.6 (br s, 2H) 3.3 (m, 5H) 1.5 (m, 2H) 1.05 (s, 9H)
N-(4-Aminophenyl)-N-(4′-fluorobiphenyl-3-sulphonyl)amino]acetic acid ethyl ester (Intermediate 60, 410 mg) was dissolved in THF (14 ml) and treated with phenyl isocyanate (114 μl). The mixture was stirred and heated at 70° C. for 6 hours. The volatiles were removed by evaporation and the residue was partitioned between saturated aqueous NaHCO3 and ethyl acetate. The organic layer was dried (Na2SO4), filtered and the filtrate was concentrated to dryness. The residue was purified by chromatography using a 10 g silica cartridge eluting with a mixture of ethyl acetate and cyclohexane (1:9) to give {N-(4′-fluorobiphenyl-3-sulphonyl)-N-[4-(3-phenylureido)phenyl]amino}acetic acid ethyl ester (280 mg).
LCMS (Method B) Rt 4.22 (M+H+) 548
1H NMR (300 MHz) (DMSO-d6) δ 8.8 (s, 1H), 8.7 (s, 1H) 8.0 (dd, 1H) 7.8 (s, 1H) 7.7-7.6 (m, 4H) 7.4-7.2 (m, 8H) 7.1 (d, 2H) 7.0 (t, 1H) 4.5 (s, 2H) 4.1 (q, 2H) 1.1 (t, 3H)
By proceeding in a similar manner the following compounds were prepared from the appropriate starting materials. The reaction may also be performed using ethyl acetate or toluene as solvent, either at room temperature or at reflux.
From 4-nitrophenyl isocyanate and 4-aminopyridine
1H NMR (400 MHz) (DMSO-d6) δ 9.7 (br s, 1H) 9.3 (br s, 1H) 8.4 (m, 2H) 8.2 (d, 2H) 7.7 (d, 2H) 7.4 (m, 2H)
From benzyl isocyanate and (4-aminophenyl)carbamic acid tert-butyl ester
LCMS (Method A) Rt 3.46 (M+H+) 342
From 4-nitrophenyl isocyanate and 3-(2-methoxyethoxy)aniline (Intermediate 70)
LCMS (Method A) Rt 3.42 (M+H+) 332
1H NMR (400 MHz) (CDCl3) δ 8.2 (d, 2H) 7.5 (d, 2H) 7.4 (br s, 1H) 7.2 (d, 1H) 7.0 (br s, 1H) 6.9 (m, 2H) 6.6 (m, 1H) 4.1 (m, 2H) 3.8 (m, 2H) 3.4 (s, 3H)
From 4-fluorobiphenyl-3-sulphonic acid N-(3-benzyloxypropyl)-N-(4-aminophenyl)amide (Intermediate 84) and phenyl isocyanate
LCMS (Method A) Rt 4.44 (M+H+) 610
From (4-aminophenyl)carbamic acid tert-butyl ester and phenyl isocyanate
LCMS (Method A) Rt 3.52 (M+H+) 328
1H NMR (300 MHz) (DMSO-d6) δ 9.2 (br s, 1H) 8.6 (br s, 1H), 8.5 (br s, 1H) 7.4 (d, 2H) 7.3 (m, 6H), 6.9 (t, 1H) 1.5 (s, 9H)
From 4′ fluorobiphenyl-3-sulphonic acid N-[(2,2-dimethyl-1,3-dioxolan-4-yl)methyl]-N-(4-aminophenyl)amide (Intermediate 78) and phenyl isocyanate
LCMS (Method B) Rt 4.25 (M+H+) 576
From 4-methyl-3-nitrophenyl isocyanate and aniline in ethyl acetate at room temperature
1H NMR (400 MHz) (DMSO-d6) δ 9.00 (s, 1H) 8.7 (s, 1H) 8.2 (s, 1H) 7.4 (m, 6H) 6.95 (s, 1H) 2.45 (s, 3H)
From 2-chloro-5-nitrophenyl isocyanate and aniline in ethyl acetate at room temperature
1H NMR (400 MHz) (DMSO-d6) δ 9.6 (s, 1H) 9.2 (s, 1H) 8.7 (s, 1H) 7.8 (d, 2H) 7.4 (d, 4H) 7.0 (s, 1H)
From 4-chloro-3-nitrophenyl isocyanate and aniline in ethyl acetate at room temperature
1H NMR (400 MHz) (DMSO-d6) δ 9.2 (s, 1H) 8.9 (s, 1H) 8.3 (s, 1H) 7.6 (m, 2H) 7.5 (d, 2H) 7.3 (t, 2H) 7.0 (t, 1H)
From 2-chloro-4-nitrophenyl isocyanate and aniline in ethyl acetate at room temperature
1H NMR (400 MHz) (DMSO-d6) δ 9.7 (s, 1H) 8.8 (s, 1H) 8.55 (d, 1H) 8.35 (s, 1H) 8.2 (d, 1H) 7.5 (d, 2H) 7.3 (t, 2H) 7.0 (t, 1H)
From 4-methyl-3-nitrophenyl isocyanate and benzylamine in ethyl acetate at room temperature
LCMS (Method A) Rt 3.46 (M+H+) 286
1H NMR (400 MHz) (DMSO-d6) □ 8.95 (s, 1H) 8.25 (s, 1H) 7.5 (d, 1H) 7.3 (m, 5H) 7.2 (m, 1H) 6.75 (t, 1H) 4.3 (d, 2H) 2.4 (s, 3H)
From 4-nitrophenyl isocyanate and 2-(pyridin-3-yl)ethylamine in ethyl acetate at room temperature.
LCMS (Method A) Rt 1.97 (M+H+) 287
From 3-methyl-4-nitrophenyl isocyanate and 2-methylaniline in ethyl acetate at room temperature.
LCMS (Method A) Rt 3.63 (M+H+) 286
From 6-nitro-2,3-dihydro-1H-indole and phenyl isocyanate in ethyl acetate at room temperature.
LCMS (Method A) Rt 3.56 (M+H+) 284
From 4-nitrophenyl isocyanate and 1-phenylethylamine in ethyl acetate at room temperature.
LCMS (Method A) Rt 3.48 (M+H+) 286
From 2-methoxy-5-nitrophenyl isocyanate and aniline in ethyl acetate at room temperature
LCMS (Method A) Rt 3.57 (M+H+) 388
From 2-(4-amino-2-methylphenyl)-5-chloro-1,3-dihydroisoindole-1,3-dione and phenyl isocyanate in ethyl acetate at reflux.
LCMS (Method A) Rt 3.8 (M+H+) 406
From N-(4-amino-2-chlorophenyl)acetamide and phenyl isocyanate in ethyl acetate at reflux
LCMS (Method B) Rt 3.03 (M+H+) 304
From 2-amino-5-nitropyridine and phenyl isocyanate in toluene at reflux
1H NMR (400 MHz) (CDCl3) δ 10.1 (s, 1H) 9.9 (s, 1H) 9.3 (s, 1H) 8.5 (d, 1H) 7.9 (d, 1H) 7.5 (d, 2H) 7.3 (t, 2H) 7.1 (t, 1H)
From 2-methoxy-4-nitrophenyl isocyanate and aniline in toluene at reflux
LCMS (Method A) Rt 3.78 (M+H+) 288
[N-(4′-Fluorobiphenyl-3-sulphonyl)-N-(4-nitrophenyl)amino]acetic acid ethyl ester (Intermediate 31, 450 mg) was dissolved in IMS (16 ml) and treated with palladium on carbon (10%, 65 mg). The reaction mixture was hydrogenated under a balloon of hydrogen at atmospheric pressure for 4 hours. The catalyst was removed by filtration through Celite under nitrogen and the filtrate was concentrated to give [N-(4-aminophenyl)-N-(4′-fluorobiphenyl-3-sulphonyl)amino]acetic acid ethyl ester (410 mg).
LCMS (Method A) Rt 3.74 (M+H+) 429
1H NMR (300 MHz) (CDCl3) δ 7.9 (s, 1H) 7.8 (dd, 1H) 7.7 (dd, 1H) 7.5 (m, 3H) 7.2 (t, 2H) 7.0 (d, 2H) 6.6 (d, 2H) 4.4 (s, 2H) 4.1 (q, 2H) 3.8 (br s, 2H) 1.2 (t, 3H)
By proceeding in a similar manner the following compounds were prepared from the appropriate starting materials:
From 4′-fluorobiphenyl-3-sulphonic acid N-methyl-N-(4-nitrophenyl)amide (Intermediate 10)
LCMS (Method A) Rt 3.58 (M+H+) 357
From N-methyl-N-(4-nitrophenyl)carbamic acid tert-butyl ester (Intermediate 20)
LCMS (Method A) Rt 2.38 (M+H+) 223
1H NMR (400 MHz) (CDCl3) δ 7.0 (d, 2H) 6.6 (d, 2H) 3.6 (br s, 2H) 3.2 (s, 3H) 1.5 (s, 9H)
From 1-(4-methyl-3-nitrophenyl)-3-phenyl urea (Intermediate 46)
1H NMR (400 MHz) δ (DMSO-d6) δ 8.4 (s, 1H) 8.1 (s, 1H) 7.3 (m, 4H) 6.7 (m, 4H) 4.7 (br s, 2H) 1.9 (s, 3H)
From 1-(4-methyl-3-nitrophenyl)-3-benzyl urea (Intermediate 50)
LCMS (Method A) Rt 2.24 (M+H+) 256
From 1-(4-nitrophenyl)-3-(pyridine-4-yl)urea (Intermediate 40)
1H NMR (400 MHz) (DMSO-d6) δ 8.9 (s, 1H) 8.3 (m, 3H) 7.4 (m, 2H) 7.2 (d, 2H) 6.5 (d, 2H) 5.8 (s, 2H)
From 1-(5-nitropyridin-2-yl)-3-phenyl urea (Intermediate 58)
1H NMR (400 MHz) (CDCl3) δ 10.4 (br s, 1H) 9.0 (s, 1H) 7.7 (s, 1H) 7.5 (d, 2H) 7.3 (t, 2H) 7.2 (d, 1H) 7.0 (m, 2H) 5.0 (br s, 2H)
From 1-(4-nitrophenyl)-3-[2-(pyridin-3-yl)ethyl]urea (Intermediate 51)
The compound used without purification or characterisation
From 1-(3-methyl-4-nitrophenyl)-3-(2-methylphenyl)urea (Intermediate 52)
LCMS (Method A) Rt 2.37 (M+H+) 256
From 6-nitro-2,3-dihydroindole-1-carboxylic acid N-phenylamide (Intermediate 53)
LCMS (Method A) Rt 1.83 & 2.01 (M+H+) 254
From 3-(2-methoxyethoxy)nitrobenzene (Intermediate 30)
LCMS (Method A) Rt 0.37 & 1.55 (M+H+) 168
1H NMR (400 MHz) (CDCl3) δ 7.1 (t, 1H) 6.3 (m, 3H) 4.1 (t, 2H) 3.7 (t, 2H) 3.6 (br s, 2H) 3.4 (s, 3H)
From 1-(4-nitrophenyl)-3-[3-(2-methoxyethoxy)phenyl]urea (Intermediate 42)
LCMS (Method A) Rt 0.37 & 2.09 (M+H+) 302
1H NMR (400 MHz) (DMSO-d6) δ 8.6 (br s 1H) 8.2 (br s, 1H) 7.2-7.0 (m, 4H) 6.9 (d, 1H) 6.5 (m, 3H) 4.8 (br s, 2H) 4.0 (t, 2H) 3.6 (t, 2H) 3.3 (s, 3H)
From 1-(2-methoxy-4-nitrophenyl)-3-phenyl urea (Intermediate 59)
1H NMR (400 MHz) (DMSO-d6) δ 9.0 (s, 1H) 7.7 (s, 1H) 7.6 (d, 1H) 7.4 (d, 2H) 7.25 (t, 2H) 6.9 (t, 1H) 6.3 (d, 1H) 6.1 (dd, 1H) 4.8 (s, 2H) 3.8 (s, 3H)
From 1-(2-methoxy-5-nitrophenyl)-3-phenyl urea (Intermediate 55)
LCMS (Method A) Rt 2.06 (M+H+) 258
From 4′-fluorobiphenyl-3-sulphonic acid N-(2-methoxy-5-nitrophenyl)amide (Intermediate 5)
LCMS (Method A) Rt 2.68 (M+H+) 373
From 4′-fluorobiphenyl-3-sulphonic acid N-(2-methoxy-4-nitrophenyl)-N-(3-tert-butoxy-propyl)amide (Intermediate 29)
1H NMR (400 MHz) (DMSO-d6) δ 7.9 (dt, 1H) 7.70-7.55 (m, 5H) 7.3 (t, 2H) 6.7 (d, 1H) 6.1 (m, 2H) 5.3 (br s, 2H) 3.5 (t, 2H) 3.3 (s, 3H) 3.25 (t, 2H) 1.5 (m, 2H) 1.1 (s, 9H)
From 4′-fluorobiphenyl-3-sulphonic acid (2-methoxy-4-nitrophenyl)amide (Intermediate 13)
1H NMR (400 MHz) (CDCl3) δ 7.8 (s, 1H) 7.6 (d, 2H) 7.4 (m, 3H) 7.35 (d, 1H) 7.1 (t, 2H) 6.5 (br s, 1H) 6.25 (dd, 1H) 6.0 (s, 1H) 3.65 (br s, 2H) 3.35 (s, 3H)
From 4′-fluorbiphenyl-3-sulphonic acid N-(2-methyl-4-nitrophenyl)-N-methylamide (Intermediate 32)
LCMS (Method A) Rt 3.56 (M+H+) 371
From 4′-fluorobiphenyl-3-sulphonic acid N-(4-nitrophenyl)-N-[(2,2-dimethyl-1,3-dioxolan-4-yl)methyl]amide (Intermediate 18)
LCMS (Method B) Rt 3.83 (M+H+) 457
From 4-nitrophenylsulphonic acid (4′-fluorobiphenyl-3-yl)amide (Intermediate 34)
LCMS (Method A) Rt 3.5 (M+H+) 343
To a solution of 1-(4-nitrophenyl)-3-(1-phenylethyl)urea (Intermediate 54, 583 mg) in ethanol (50 ml) was added SnCl2.2H2O (4.27 g). The mixture was stirred and heated at 70° C. overnight. The reaction mixture was cooled to room temperature, poured onto a mixture of ice and water and then diluted with aqueous NaOH. The mixture was extracted with ethyl acetate and the organic layer was dried (MgSO4) and filtered. The volatiles were removed by evaporation to give 1-(4-aminophenyl)-3-(1-phenylethyl)urea (156 mg).
LCMS (Method A) Rt 2.06 (M+H+) 256
By proceeding in a similar manner the following compounds were prepared from the appropriate starting materials:
From 1-(2-chloro-5-nitrophenyl)-3-phenyl urea (Intermediate 47)
LCMS (Method A) Rt 2.75 (M+H+) 262
From 1-(4-chloro-3-nitrophenyl)-3-phenyl urea (Intermediate 48)
LCMS (Method A) Rt 3.23 (M+H+) 2.62
From 1-(2-chloro-4-nitrophenyl)-3-phenyl urea (Intermediate 49)
1H NMR (400 MHz) (DMSO-d6) □ 8.9 (s, 1H) 7.8 (s, 1H) 7.5 (d, 1H) 7.4 (d, 2H) 7.25 (t, 2H) 6.9 (t, 1H) 6.65 (d, 1H) 6.5 (dd, 1H) 5.2 (s, 2H)
From 4′-fluorobiphenyl-3-sulphonic acid N-(4-nitrophenyl)-N-(3-benzyloxypropyl)amide (Intermediate 28)
LCMS (Method B) Rt 4.23 (M+H+) 491
To a suspension of N-[2-chloro-4-(3-phenylureido)phenyl]acetamide (Intermediate 57, 690 mg) in ethanol (10 ml) was added HCl (37%, 10 ml). The reaction mixture was heated at reflux for 2 hours, then cooled and poured directly onto a SCX-2 column. It was eluted with DCM, then MeOH and then with a mixture DCM and 2M ammonia in MeOH (9:1) to give 1-(4-amino-3-chlorophenyl)-3-phenyl urea as a yellow solid (250 mg).
LCMS (Method B) Rt 2.92 (M+H+) 262
To a solution of 4-fluorophenyl boronic acid (750 mg) in pyridine (67 ml) was added copper acetate (1.95 g) and pyrazole (729 mg). The mixture was stirred in an open reaction vessel at 40° C. overnight. The pyridine was removed by evaporation and the residue was partitioned between water and ethyl acetate. The organic layer was washed with water, dried (Na2SO4) and filtered. The volatiles were removed by evaporation to give 1-(4-fluorophenyl)-1H-pyrazole (816 mg).
LCMS (Method A) Rt 3.18 (M+H+) 163
1H NMR (300 MHz) (DMSO-d6) δ 8.5 (s, 1H) 7.9 (m, 2H) 7.7 (s, 1H) 7.3 (m, 2H) 6.5 (s, 1H)
1-(4-Fluorophenyl)-1H-pyrazole (Intermediate 86, 300 mg) was dissolved in chloroform (24 ml) and chlorosulphonic acid (1.23 ml) was added. The mixture was heated under reflux for 3 hours then the volatiles were removed by evaporation. The residue was treated with thionyl chloride (9.2 ml) and DMF (9 drops) and the mixture was stirred and heated at 100° C. for 2 hours, then cooled to room temperature. The volatiles were again removed by evaporation and the residue was treated with toluene and re-evaporated. The residue was then partitioned between water and ethyl acetate and the organic layer was dried (Na2SO4) and filtered. The volatiles were removed by evaporation to give 1-(4-fluorophenyl)-1H-pyrazole-4-sulphonyl chloride as a pale brown oil (480 mg).
1H NMR (300 MHz) (DMSO-d6) δ 8.5 (s, 1H) 7.9 (m, 2H) 7.7 (s, 1H) 7.3 (t, 2H)
By proceeding in a similar manner the following compounds were prepared from the appropriate starting materials:
From 3,5-dimethyl-1-(pyridin-2-yl)-1H-pyrazole
LCMS (Method A) Rt 3.82 (M+H+) 272
1H NMR (300 MHz) (DMSO-d6) δ 8.5 (dd, 1H) 8.0 (m, 1H) 7.7 (d, 1H) 7.4 (m, 1H), 2.7 (s, 3H) 2.3 (s, 3H)
From 1-(pyridine-2-yl)-1H-pyrazole (Intermediate 93)
LCMS (Method A) Rt 3.64 (M+H+) 244
From 1-(5-fluoropyridin-2-yl)-1H-pyrazole (Intermediate 92)
1H NMR (300 MHz) (DMSO-d6) δ 8.5 (s, 1H) 8.3 (s, 1H) 7.9 (m, 2H) 7.6 (s, 1H)
From 2-(4-fluorophenyl)-1-methyl-1H-imidazole
LCMS (Method B) Rt 3.46 (M+H+) 275
1H NMR (400 MHz) (DMSO-d6) δ 7.75 (s, 1H) 7.65 (m, 2H) 7.20 (m, 2H) 3.9 (s, 3H)
A suspension of copper (I) iodide (48 mg), L-proline (59 mg), potassium carbonate (730 mg), 2-bromo-5-fluoropyridine (500 mg) and pyrazole (175 mg) in DMSO (3.3 ml) was heated in the microwave for 2 hours at 140° C. The resultant mixture was partitioned between water and ethyl acetate and the organic layer was dried (MgSO4) and filtered. The volatiles were removed by evaporation and the residue was purified by chromatography on a 5 g silica cartridge eluting with initially cyclohexane increasing the polarity to a mixture of ethyl acetate and cyclohexane (1:20) to give 1-(5-fluoropyrid-2-yl)-1H-pyrazole (200 mg).
LCMS (Method B) Rt 2.94 (M+H+) 164
1H NMR (300 MHz) (DMSO-d6) δ 8.6 (d, 1H) 8.5 (t, 1H) 8.0 (m, 2H) 7.8 (d, 1H) 6.6 (dd, 1H)
By proceeding in a similar manner the following compound was prepared from the appropriate starting materials:
From 2-bromopyridine and pyrazole
1H NMR (300 MHz) (DMSO-d6) δ 8.6 (s, 1H) 8.5 (m, 1H) 8.0 (m, 2H) 7.8 (s, 1H) 7.3 (t, 1H) 6.6 (s, 1H)
4′-Fluorobiphenyl-3-sulphonic acid (4-aminophenyl)amide (Intermediate 22, 50 mg) was dissolved in THF (2 ml) and treated with NaOH (1M, 300 μl) and 4-methoxyphenyl isocyanate (29 μl). After stirring at room temperature for 20 hours, the THF was removed by evaporation and the residue was acidified to pH 5 and extracted with ethyl acetate. The organic layer was dried (Na2SO4), filtered and the volatiles were removed by evaporation. The residue was purified by HPLC eluting with a mixture of water and acetonitrile (each containing 0.1% of formic acid) from 20 to 98% acetonitrile over 30 minutes to give 4′-fluorobiphenyl-3-sulphonic acid {4-[3-(4-methoxyphenyl)ureido]-phenyl}amide as a white solid (62 mg).
LCMS (Method C) Rt 11.30 (M+H+) 492
1H NMR (400 MHz) (DMSO-d6) δ 10.0 (br s, 1H) 8.5 (br s, 1H) 8.4 (br s, 1H) 7.9 (m, 2H) 7.7 (m, 4H) 7.3 (m, 6H) 7.0 (d, 2H) 6.8 (d, 2H) 3.7 (s, 3H)
By proceeding in a similar manner the following compounds were prepared from the appropriate starting materials:
From 4′-fluorobiphenyl-3-sulphonic acid (4-aminophenyl)amide (Intermediate 22) and 2-chlorophenyl isocyanate
LCMS (Method C) Rt 12.34 (M+H+) 496
1H NMR (400 MHz) (DMSO-d6) δ 10.0 (br s, 1H) 9.3 (s, 1H) 8.3 (s, 1H) 8.1 (d, 1H) 7.9 (m, 2H) 7.7 (m, 4H) 7.4 (d, 1H) 7.3 (m, 5H) 7.0 (m, 3H)
From 4′-fluorobiphenyl-3-sulphonic acid (4-aminophenyl)amide (Intermediate 22) and 2-methoxyphenyl isocyanate
LCMS (Method C) Rt 11.94 (M+H+) 492
1H NMR (400 MHz) (DMSO-d6) δ 10.0 (br s, 1H) 9.2 (s, 1H) 8.2 (s, 1H) 8.1 (d, 1H) 7.9 (m, 2H) 7.7 (m, 4H) 7.3 (m, 4H) 7.0 (m, 3H) 6.9 (m, 2H) 3.9 (s, 3H)
4′-Fluorobiphenyl-3-sulphonic acid (4-aminophenyl)amide (Intermediate 22, 50 mg) was dissolved in ethyl acetate (2.5 ml) and then treated with 2-methylphenyl isocyanate (22 μl). The resultant mixture was stirred and heated at 85° C. overnight. The reaction mixture was cooled to room temperature and partitioned between aqueous citric acid (10%) and ethyl acetate. The organic layer was dried (Na2SO4), filtered and the volatiles were removed by evaporation. The residue was purified by HPLC eluting with a mixture of water and acetonitrile (each containing 0.1% formic acid) from 30 to 98% acetonitrile over 20 minutes to give 4′-fluorobiphenyl-3-sulphonic acid {4-[3-(2-methylphenyl)ureido]phenyl}amide as a white solid (15 mg).
LCMS (Method C) Rt 11.78 (M+H+) 476
1H NMR (400 MHz) (CDCl3) δ 7.85 (s, 1H) 7.7 (m, 2H) 7.5-7.4 (m, 4H) 7.3-7.2 (m, 5H) 7.1 (m, 2H) 7.0 (m, 2H) 6.3 (br s, 2H) 6.0 (s, 1H) 2.3 (s, 3H)
By proceeding in a similar manner the following compounds were prepared from the appropriate starting materials. Alternative solvents such as THF or toluene may be used and the reaction may be carried out at room temperature or at reflux.
From 4′-fluorobiphenyl-3-sulphonic acid (5-amino-2-methoxyphenyl)amide (Intermediate 74) and phenyl isocyanate.
LCMS (Method C) Rt 11.78 (M+H+) 492
1H NMR (400 MHz) (DMSO-d6) δ 9.5 (s, 1H) 8.6 (s, 1H) 8.5 (s, 1H) 8.0 (s, 1H) 7.9 (d, 1H) 7.7 (m, 3H) 7.6 (t, 1H) 7.5 (s, 1H) 7.4 (d, 2H) 7.3 (m, 4H) 7.1 (dd, 1H) 6.9 (t, 1H) 6.8 (d, 1H) 3.5 (s, 3H)
From 4′-fluorobiphenyl-3-sulphonic acid (4-aminophenyl)amide (Intermediate 22) and 4-cyanophenyl isocyanate in THF at reflux
LCMS (Method C) Rt 11.36 (M+H+) 487
1H NMR (400 MHz) (DMSO-d6) δ 10.1 (s, 1H) 9.2 (s, 1H) 8.8 (s, 1H) 7.9 (m, 2H) 7.7-7.6 (m, 8H) 7.3 (m, 4H) 7.0 (d, 2H)
From 4′-fluorobiphenyl-3-sulphonic acid N-(4-aminophenyl)-N-methylamide (Intermediate 61) and 4-pyridyl isocyanate in ethyl acetate at room temperature
LCMS (Method C) Rt 7.84 (M+H+) 477
1H NMR (400 MHz) (CD3OD) δ 8.4 (br s, 2H) 7.9 (d, 1H) 7.7 (m, 2H) 7.6 (m, 2H) 7.5 (m, 3H) 7.4 (d, 2H) 7.2 (t, 2H) 7.1 (d, 2H) 3.2 (s, 3H)
From 4′-fluorobiphenyl-3-sulphonic acid N-(4-amino-2-methylphenyl)-N-methylamide (Intermediate 77) and phenyl isocyanate in ethyl acetate at room temperature
LCMS (Method C) Rt 12.57 (M+H+) 490
1H NMR (400 MHz) (DMSO-d6) δ 8.6 (s, 1H) 8.5 (s, 1H) 8.0 (m, 1H) 7.8 (t, 3H) 7.7 (m, 2H) 7.3 (d, 2H) 7.2 (m, 6H) 7.0 (s, 1H) 6.9 (t, 1H) 3.1 (s, 3H) 2.2 (s, 3H)
From 4′-fluorobiphenyl-3-sulphonic acid (4-amino-2-methoxyphenyl)amide (Intermediate 76) and phenyl isocyanate in toluene at reflux.
LCMS (Method C) Rt 11.70 (M+H+) 492
1H NMR (400 MHz) (DMSO-d6) δ 9.4 (br s, 1H) 8.8 (s, 1H) 8.7 (s, 1H) 7.9 (m, 2H) 7.7 (m, 4H) 7.4 (d, 2H) 7.3 (m, 4H) 7.2 (s, 1H) 7.1 (d, 1H) 7.0 (t, 1H) 6.9 (d, 1H) 3.3 (s, 3H)
From 4-aminophenylsulphonic acid (4′-fluorobiphenyl-3-yl)amide (Intermediate 79) and phenyl isocyanate in toluene at reflux.
LCMS (Method C) Rt 11.81 (M+H+) 462
1H NMR (400 MHz) (DMSO-d6) δ 10.2 (br s, 1H) 9.0 (s, 1H) 8.7 (s, 1H) 7.7 (d, 2H) 7.6-7.5 (m, 4H) 7.4 (br d, 2H) 7.3-7.2 (m, 7H) 7.0 (dt, 1H) 6.9 (br t, 1H).
To a solution of 2,4-dichlorophenylsulphonyl chloride (60 mg) in pyridine (1.5 ml) was added 1-(4-methylaminophenyl)-3-phenyl urea (Intermediate 25, 59 mg). The reaction mixture was stirred for 4 hours at room ternperature and then the volatiles were removed by evaporation. The residue was partitioned between saturated aqueous NaHCO3 and ethyl acetate. The organic layer was dried (Na2SO4), filtered and volatiles were removed by evaporation. The residue was purified by HPLC eluting with a mixture of water and acetonitrile (each containing 0.1% formic acid) from 20 to 98% over 25 minutes to give 2,4-dichlorophenyl sulphonic acid N-methyl-N-[4-(3-phenylureido)phenyl]amide (40 mg).
LCMS (Method C) Rt 12.08 (M+H+) 450
1H NMR (400 MHz) (DMSO-d6) δ 8.8 (br s, 1H) 8.7 (br s, 1H) 7.9 (s, 1H) 7.8 (d, 1H) 7.6 (d, 1H) 7.4 (m, 4H) 7.3 (t, 2H) 7.1 (d, 2H) 7.0 (t, 1H) 3.3 (s, 3H)
By proceeding in a similar manner the following compounds were prepared from the appropriate starting materials. The reactions may be performed using alternative solvents such as DCM or NMP in the presence of a base such as pyridine or N,N-diisopropyl-N-ethylamine.
From 4′-fluorobiphenyl-3-sulphonyl chloride and 1-(4-methylaminophenyl)-3-phenyl urea (Intermediate 25)
LCMS (Method C) Rt 12.47 (M+H+) 476
1H NMR (400 MHz) (DMSO-d6) δ 8.8 (s, 1H) 8.7 (s, 1H) 8.0 (d, 1H) 7.7 (m, 3H) 7.6 (s, 1H) 7.5 (d, 1H) 7.4 (m, 4H) 7.3 (m, 4H) 7.0 (m, 3H) 3.2 (s, 3H)
From 3-difluoromethoxyphenylsulphonyl chloride and 1-(4-methylaminophenyl)-3-phenyl urea (Intermediate 25)
LCMS (Method C) Rt 11.35 (M+H+) 448
1H NMR (400 MHz) (DMSO-d6) δ 8.9 (s, 1H) 8.7 (s, 1H) 7.7 (t, 1H) 7.5 (dd, 1H) 7.4 (m, 5H) 7.3 (m, 4H) 7.0 (m, 3H) 3.1 (s, 3H)
From 2-chloro-5-trifluoromethylphenylsuphonyl chloride and 1-(4-methylaminophenyl)-3-phenyl urea (Intermediate 25)
LCMS (Method C) Rt 12.11 (M+H+) 484
1H NMR (400 MHz) (DMSO-d6) δ 8.8 (s, 1H) 8.7 (s, 1H) 8.1 (d, 1H) 8.0 (d, 1H) 7.9 (s, 1H) 7.4 (m, 4H) 7.3 (t, 2H) 7.1 (d, 2H) 7.0 (t, 1H) 3.3 (s, 3H)
From 4′ fluorobiphenylsulphonyl chloride and 1-(4-aminophenyl)-3-(pyrid-4-yl)urea (Intermediate 65)
LCMS (Method C) Rt 7.42 (M+H+) 463
1H NMR (400 MHz) (DMSO-d6) δ 10.0 (br s, 1H) 9.0 (s, 1H) 8.8 (s, 1H) 8.3 (d, 2H) 7.9 (m, 2H) 7.7 (m, 4H) 7.3 (m, 6H) 7.0 (d, 2H)
From 4′-fluorobiphenyl-3-sulphonyl chloride and 1-(4-aminophenyl)-3-[2-(pyridin-3-yl)ethyl]urea (Intermediate 67)
LCMS (Method C) Rt 7.26 (M+H+) 491
1H NMR (400 MHz) (DMSO-d6) δ 9.9 (s, 1H) 8.4 (m, 3H) 7.9 (m, 2H) 7.7 (m, 5H) 7.3 (m, 3H) 7.2 (d, 2H) 6.9 (d, 2H) 6.1 (t, 1H) 3.3 (m, 2H) 2.7 (t, 2H)
From 4′-fluorobiphenyl-3-sulphonyl chloride and 1-(3-Amino-4-chlorophenyl)-3-phenyl urea (Intermediate 82)
LCMS (Method C) Rt 12.38 (M+H+) 496
1H NMR (400 MHz) (DMSO-d6) δ 10.0 (s, 1H) 8.9 (s, 1H) 8.6 (s, 1H) 8.0 (s, 1H) 7.9 (d, 1H) 7.8 (d, 1H) 7.7 (m, 4H) 7.4 (d, 2H) 7.3 (m, 5H) 7.2 (d, 1H) 7.0 (t, 1H)
From 4′-fluorobiphenyl-3-sulphonyl chloride and 1-(3-amino-4-methylphenyl)-3-benzyl urea (Intermediate 64)
LCMS (Method C) Rt 11.69 (M+H+) 490
1H NMR (400 MHz) (DMSO-d6) δ 9.5 (s, 1H) 8.5 (s, 1H) 7.9 (m, 2H) 7.7 (m, 4H) 7.3 (m, 8H) 7.2 (d, 1H) 6.9 (d, 1H) 6.5 (t, 1H) 4.3 (d, 2H) 1.9 (s, 3H)
From 4′-fluorobiphenyl-3-sulphonyl chloride and 1-(4-amino-3-methylphenyl)-3-(2-methylphenyl)urea (Intermediate 68)
LCMS (Method C) Rt 12.14 (M+H+) 490
1H NMR (400 MHz) (DMSO-d6) δ 9.6 (br s, 1H) 9.0 (s, 1H) 7.9 (m, 2H) 7.8 (m, 2H) 7.7 (m, 4H) 7.3 (m, 3H) 7.2 (m, 3H) 7.0 (d, 1H) 6.9 (t, 1H) 2.2 (s, 3H) 1.9 (s, 3H)
From 4′fluorobiphenyl-3-sulphonyl chloride and 6-amino-2,3-dihydroindole-1-carboxylic acid N-phenylamide (Intermediate 69)
LCMS (Method C) Rt 11.98 (M+H+) 488
1H NMR (400 MHz) (DMSO-d6) δ 10.2 (br s, 1H) 8.4 (s, 1H) 8.0 (s, 1H) 7.9 (m, 2H) 7.7 (m, 3H) 7.6 (t, 1H) 7.5 (d, 2H) 7.3 (t, 2H) 7.2 (t, 2H) 7.0 (m, 2H) 6.7 (d, 1H) 4.1 (t, 2H) 3.1 (t, 2H)
From 4′-fluorobiphenyl-3-sulphonyl chloride and 1-(4-amino-3-methylphenyl)-3-phenyl urea (Intermediate 27)
LCMS (Method C) Rt 11.45 (M+H+) 476
1H NMR (400 MHz) (DMSO-d6) δ 9.4 (s, 1H) 8.65 (s, 1H) 8.6 (s, 1H) 7.9 (dd, 1H) 7.8 (s, 1H) 7.6 (m, 4H) 7.4 (d, 2H) 7.3 (m, 5H) 7.2 (dd, 1H) 7.0 (t, 1H) 6.8 (d, 1H) 2.0 (d, 3H)
From 4′-fluorobiphenylsulphonyl chloride and 1-(4-aminophenyl)-3-(1-phenylethyl)urea (Intermediate 80)
LCMS (Method C) Rt 11.55 (M+H+) 490
1H NMR (400 MHz) (DMSO-d6) δ 9.9 (br s, 1H) 8.3 (s, 1H) 7.9 (m, 2H) 7.6 (m, 4H) 7.3 (m, 6H) 7.2 (m, 3H) 6.9 (d, 2H) 6.6 (d, 1H) 4.8 (dq, 1H) 1.4 (d, 3H)
From 4′-fluorobiphenylsulphonyl chloride and 1-(4-aminophenyl)-3-[3-(2-methyoxy-ethoxy)phenyl]urea (Intermediate 71)
LCMS (Method C) Rt 11.32 (M+H+) 536
1H NMR (400 MHz) (DMSO-d6) δ 10.0 (s, 1H) 8.6 (s, 1H) 8.55 (s, 1H) 7.9 (m, 2H) 7.7 (m, 4H) 7.3 (m, 4H) 7.1 (m, 2H) 7.0 (d, 2H) 6.9 (d, 1H) 6.5 (d, 1H) 4.0 (t, 2H) 3.6 (t, 2H) 3.3 (s, 3H)
From 1-(4-fluorophenyl)-3,5-dimethyl-1H-pyrazole-4-sulphonyl chloride and 1-(4-amino-phenyl)-3-phenyl urea (Intermediate 23)
LCMS (Method C) Rt 10.52 (M+H+) 480
1H NMR (400 MHz) (DMSO-d6) δ 9.8 (s, 1H) 8.6 (s, 2H) 7.5 (m, 2H) 7.4 (d, 2H) 7.3 (m, 4H) 7.2 (t, 2H) 7.0 (m, 3H) 2.2 (s, 6H)
From 1-(4-fluorophenyl)-3,5-dimethyl-1H-pyrazole-4-sulphonyl chloride and 1-(4-methyl-aminophenyl)-3-phenyl urea (Intermediate 25)
LCMS (Method C) Rt 11.44 (M+H+) 494
1H NMR (400 MHz) (DMSO-d6) δ 8.8 (s, 1H) 8.7 (s, 1H) 7.6 (m, 2H) 7.4 (m, 6H) 7.3 (t, 2H) 7.2 (d, 2H) 7.0 (t, 1H) 3.1 (s, 3H) 2.1 (s, 3H) 2.0 (s, 3H)
From 5-(1-methyl-3-trifluoromethyl-1H-pyrazol-5-yl)thiophene-2-sulphonyl chloride and 1-(5-amino-2-chlorophenyl)-3-phenyl urea (Intermediate 81)
LCMS (Method C) Rt 12.12 (M+H+) 556
1H NMR (400 MHz) (DMSO-d6) δ 10.7 (br s, 1H) 9.4 (s, 1H) 8.3 (s, 1H) 8.1 (s, 1H) 7.7 (d, 1H) 7.6 (d, 1H) 7.5 (d, 2H) 7.4 (d, 1H) 7.3 (t, 2H) 7.2 (s, 1H) 7.0 (t, 1H) 6.8 (dd, 1H) 4.0 (s, 3H)
From 1-(4-fluorophenyl)-1H-pyrazole-4-sulphonyl chloride (Intermediate 87) and 1-(4-methylaminophenyl)-3-phenyl urea (Intermediate 25)
LCMS (Method C) Rt 11.34 (M+H+) 466
1H NMR (400 MHz) (DMSO-d6) δ 9.0 (s, 1H) 8.8 (s, 1H) 8.7 (s, 1H) 8.0 (m, 2H) 7.8 (s, 1H) 7.4 (m, 6H) 7.3 (t, 2H) 7.1 (d, 2H) 7.0 (t, 1H) 3.2 (s, 3H)
From 1-(5-trifluoromethylpyridin-2-yl)-1H-pyrazole-4-sulphonyl chloride and 1-(4-methyl-aminophenyl)-3-phenyl urea (Intermediate 25)
LCMS (Method C) Rt 12.16 (M+H+) 517
1H NMR (400 MHz) (DMSO-d6) δ 9.0 (s, 1H) 8.9 (s, 1H) 8.8 (s, 1H) 8.7 (s, 1H) 8.5 (dd, 1H) 8.2 (dd, 1H) 8.0 (s, 1H) 7.4 (dd, 4H) 7.3 (t, 2H) 7.1 (d, 2H) 7.0 (t, 1H) 3.2 (s, 3H)
From and 3,5-dichlorophenylsulphonyl chloride and 1-(4-methylaminophenyl)-3-phenyl urea (Intermediate 25)
LCMS (Method C) Rt 12.43 (M+H+) 450
1H NMR (400 MHz) (DMSO-d6) δ 8.8 (s, 1H) 8.7 (s, 1H) 8.1 (s, 1H) 7.4 (m, 6H) 7.3 (t, 2H) 7.0 (d, 2H) 6.9 (t, 1H) 3.2 (s, 3H)
From 5-(oxazol-5-yl)thiophene-2-sulphonyl chloride and 1-(4-methylaminophenyl)-3-phenyl urea (Intermediate 25)
LCMS (Method C) Rt 10.52 (M+H+) 455
1H NMR (400 MHz) (DMSO-d6) δ 8.8 (s, 1H) 8.7 (s, 1H) 8.5 (s, 1H) 7.8 (s, 1H) 7.6 (d, 1H) 7.5 (d, 1H) 7.4 (m, 4H) 7.3 (t, 2H) 7.1 (d, 2H) 7.0 (t, 1H) 3.2 (s, 3H)
From 5-methyl-1-phenyl-1H-pyrazole-4-sulphonyl chloride and 1-(4-methylaminophenyl)-3-phenyl urea (Intermediate 25) using pyridine in DCM
LCMS (Method C) Rt 11.10 (M+H+) 462
1H NMR (400 MHz) ((DMSO-d6) δ 8.8 (s, 1H) 8.7 (s, 1H) 7.8 (s, 1H) 7.55 (m, 5H) 7.45 (m, 4H) 7.3 (t, 2H) 7.1 (d, 2H) 6.95 (t, 1H) 3.1 (s, 3H) 1.9 (s, 3H)
From 5-(pyridin-2-yl)thiophene-2-sulphonyl chloride and 1-(4-aminophenyl)-3-phenyl urea (Intermediate 23)
LCMS (Method C) Rt 10.31 (M+H+) 450
1H NMR (400 MHz) (DMSO-d6) δ 10.2 (br s, 1H) 8.8 (2s, 2H) 8.7 (d, 1H) 8.0 (d, 1H) 7.9 (t, 1H) 7.8 (d, 1H) 7.5 (d, 1H) 7.4 (m, 5H) 7.3 (t, 2H) 7.1 (d, 2H) 6.9 (t, 1H)
From 5-(1-methyl-3-trifluoromethyl-1H-pyrazol-5-yl)thiophene-2-sulphonyl chloride and 1-(4-aminophenyl)-3-phenyl urea (Intermediate 23)
LCMS (Method C) Rt 11.36 (M+H+) 522
1H NMR (400 MHz) (DMSO-d6) δ 10.6 (br s, 1H) 8.65 (s, 1H) 8.6 (s, 1H) 7.5 (dd, 2H) 7.4 (dd, 4H) 7.3 (t, 2H) 7.2 (s, 1H) 7.1 (d, 2H) 6.9 (t, 1H) 4.0 (s, 3H)
From 5-(pyridin-2-yl)thiophene-2-sulphonyl chloride and 1-(4-methylaminophenyl)-3-phenyl urea (Intermediate 25)
LCMS (Method C) Rt 11.27 (M+H+) 465
1H NMR (400 MHz) (DMSO-d6) δ 8.8 (s, 1H) 8.7 (s, 1H) 8.6 (d, 1H) 8.1 (d, 1H) 7.9 (m, 2H) 7.5 (m, 6H) 7.3 (t, 2H) 7.1 (d, 2H) 7.0 (t, 1H) 3.2 (s, 3H)
From 3,5-dimethyl-1-(pyridin-2-yl)-1H-pyrazole-4-sulphonyl chloride (Intermediate 88) and 1-(4-methylaminophenyl)-3-phenyl urea (Intermediate 25)
LCMS (Method C) Rt 10.90 (M+H+) 477
1H NMR (400 MHz) (DMSO-d6) δ 8.8 (s, 1H) 8.7 (s, 1H) 8.6 (dd, 1H) 8.1 (t, 1H) 7.8 (d, 1H) 7.5 (m, 5H) 7.3 (t, 2H) 7.2 (d, 2H) 7.0 (t, 1H) 3.2 (s, 3H) 2.4 (s, 3H) 2.0 (s, 3H)
From 1-(pyridin-2-yl)-1H-pyrazole-2-sulphonyl chloride (Intermediate 89) and 1-(4-methylaminophenyl)-3-phenyl urea (Intermediate 25)
LCMS (Method C) Rt 10.89 (M+H+) 449
1H NMR (400 MHz) (DMSO-d6) δ 8.8 (s, 2H) 8.7 (s, 1H) 8.55 (d, 1H) 8.1 (m, 1H) 8.0 (d, 1H) 7.9 (s, 1H) 7.5 (m, 1H) 7.4 (m, 4H) 7.3 (t, 2H) 7.1 (d, 2H) 7.0 (t, 1H) 3.2 (s, 3H)
From 5-(isoxazol-5-yl)thiophene-2-sulphonyl chloride and 1-(4-methylaminophenyl)-3-phenyl urea (Intermediate 25)
LCMS (Method C) Rt 11.06 (M+H+) 455
1H NMR (400 MHz) (DMSO-d6) δ 8.8 (s, 1H) 8.75 (s, 1H) 8.7 (s, 1H) 7.8 (d, 1H) 7.6 (d, 1H) 7.5 (m, 4H) 7.3 (t, 2H) 7.1 (m, 3H) 7.0 (t, 1H) 3.2 (s, 3H)
From 3,5-dichlorophenyl sulphonyl chloride and 1-(4-aminophenyl)-3-phenyl urea (Intermediate 23)
LCMS (Method C) Rt 11.45 (M+H+) 436
1H NMR (400 MHz) (DMSO-d6) δ 10.2 (br s, 1H) 8.65 (s, 1H) 8.6 (s, 1H) 8.0 (s, 1H) 7.65 (s, 2H) 7.45 (d, 2H) 7.4 (d, 2H) 7.3 (t, 2H) 7.0 (m, 3H)
From 4′-fluorobiphenyl-3-sulphonic acid and 1-(4-amino-2-methoxyphenyl)-3-phenyl urea (Intermediate 72)
LCMS (Method C) Rt 11.88 (M+H+) 492
1H NMR (400 MHz) (DMSO-d6) δ 10.0 (s, 1H) 9.2 (s, 1H) 8.1 (s, 1H) 7.9 (m, 3H) 7.7 (m, 4H) 7.4 (d, 2H) 7.3 (t, 2H) 7.2 (t, 2H) 7.0 (t, 1H) 6.8 (s, 1H) 6.6 (d, 1H) 3.7 (s, 3H)
From 2-chloro-4-methylphenylsulphonyl chloride and 1-(4-methylaminophenyl)-3-phenyl urea (Intermediate 25)
LCMS (Method C) Rt 11.66 (M+H+) 430
1H NMR (400 MHz) (DMSO-d6) δ 8.8 (s, 1H) 8.7 (s, 1H) 7.7 (d, 1H) 7.5 (s, 1H) 7.4 (m, 4H) 7.3 (m, 3H) 7.1 (d, 2H) 7.0 (t, 1H) 3.3 (s, 3H) 2.4 (s, 3H)
From 2,3-dichlorophenylsulphonyl chloride and 1-(4-methylaminophenyl)-3-phenyl urea (Intermediate 25)
LCMS (Method C) Rt 11.86 (M+H+) 450
1H NMR (400 MHz) (DMSO-d6) δ 8.8 (s, 1H) 8.7 (s, 1H) 8.0 (d, 1H) 7.8 (d, 1H) 7.5 (t, 1H) 7.4 (m, 4H) 7.3 (t, 2H) 7.1 (d, 2H) 7.0 (t, 1H) 3.3 (s, 3H)
From 5-(1-methyl-3-trifluoromethyl-1H-pyrazole-5-yl)thiophene-2-sulphonyl chloride and 1-(4-methylaminophenyl)-3-phenyl urea (Intermediate 25)
LCMS (Method C) Rt 12.20 (M+H+) 536
1H NMR (400 MHz) (DMSO-d6) δ 8.9 (s, 1H) 8.7 (s, 1H) 7.7 (d, 1H) 7.6 (d, 1H) 7.4 (d, 4H) 7.3 (t, 2H) 7.2 (s, 1H) 7.1 (d, 2H) 6.9 (t, 1H) 4.0 (s, 3H) 3.2 (s, 3H)
From 5-(5-trifluoromethylisoxazol-3-yl)thiophene-2-sulphonyl chloride and 1-(4-methylaminophenyl)-3-phenyl urea (Intermediate 25)
LCMS (Method C) Rt 12.70 (M+H+) 523
1H NMR (400 MHz) (DMSO-d6) δ 8.8 (s, 1H) 8.6 (s, 1H) 8.2 (s, 1H) 8.0 (d, 1H) 7.7 (d, 1H) 7.5 (m, 4H) 7.3 (t, 2H) 7.1 (d, 2H) 7.0 (t, 1H) 3.2 (s, 3H)
From 5-(1-methyl-5-trifluoromethyl-1H-pyrazol-3-yl)thiophene-2-sulphonyl chloride and 1-(4-methylaminophenyl)-3-phenyl urea (Intermediate 25)
LCMS (Method C) Rt 12.49 (M+H+) 536
1H NMR (400 MHz) (DMSO-d6) δ 8.8 (s, 1H) 8.7 (s, 1H) 7.7 (d, 1H) 7.6 (s, 1H) 7.5 (m, 5H) 7.3 (t, 2H) 7.1 (d, 2H) 7.0 (t, 1H) 4.0 (s, 3H) 3.2 (s, 3H)
From 5-(2-methylthiazol-4-yl)thiophene-2-sulphonyl chloride and 1-(4-methylaminophenyl)-3-phenyl urea (Intermediate 25)
LCMS (Method C) Rt 11.61 (M+H+) 485
1H NMR (400 MHz) (DMSO-d6) δ 8.8 (s, 1H) 8.7 (s, 1H) 8.1 (s, 1H) 7.65 (d, 1H) 7.45 (m, 5H) 7.3 (t, 2H) 7.1 (d, 2H) 6.95 (t, 1H) 3.2 (s, 3H) 2.7 (s, 3H)
From 5-(5-methyl-1,3,4-oxadiazol-2-yl)thiophene-2-sulphonyl chloride and 1-(4-methylaminophenyl)-3-phenyl urea (Intermediate 25)
LCMS (Method C) Rt 10.23 (M+H+) 470
1H NMR (400 MHz) (DMSO-d6) δ 8.8 (s, 1H) 8.7 (s, 1H) 7.9 (d, 1H) 7.6 (d, 1H) 7.5 (m, 4H) 7.3 (t, 2H) 7.1 (d, 2H) 7.0 (t, 1H) 3.2 (s, 3H) 2.6 (s, 3H)
From 1-(5-fluoropyridin-2-yl)-1H-pyrazole-4-sulphonyl chloride (Intermediate 90) and 1-(4-methylaminophenyl)-3-phenyl urea (Intermediate 25)
LCMS (Method C) Rt 11.20 (M+H+) 467
1H NMR (400 MHz) (DMSO-d6) δ 8.8 (s, 1H) 8.75 (s, 1H) 8.7 (s, 1H) 8.6 (s, 1H) 8.1 (d, 2H) 7.9 (s, 1H) 7.5 (m, 4H) 7.3 (t, 2H) 7.1 (d, 2H) 6.9 (t, 1H) 3.1 (s, 3H)
From 4′-fluorobiphenyl-3-sulphonyl chloride and 1-(4-aminophenyl)-3-phenyl urea (Intermediate 23) using N,N-diisopropyl-N-ethylamine in DCM
LCMS (Method C) Rt 11.26 (M+H+) 462
1H NMR (400 MHz) (DMSO-d6) δ 10.0 (br s, 1H) 8.6 (s, 1H) 8.55 (s, 1H) 7.9 (m, 2H) 7.7 (m, 4H) 7.4 (d, 2H) 7.3 (m, 6H) 7.0 (d, 2H) 7.95 (t, 1H)
From 3-tert-butylphenylsulphonyl chloride and 1-(4-methylaminophenyl)-3-phenyl urea (Intermediate 25) using pyridine in DCM
LCMS (Method C) Rt 12.50 (M+H+) 438
1H NMR (400 MHz) (DMSO-d6) δ 8.9 (s, 1H) 8.8 (s, 1H) 7.8 (d, 1H) 7.6 (t, 1H) 7.4 (m, 5H) 7.3 (m, 3H) 6.9 (m, 3H) 3.05 (s, 3H) 1.2 (s, 9H)
From 3-cyanophenylsuphonyl chloride and 1-(4-methylaminophenyl)-3-phenyl urea (Intermediate 25) using pyridine in DCM
LCMS (Method C) Rt 10.70 (M+H+) 407
1H NMR (400 MHz) (DMSO-d6) δ 8.9 (s, 1H) 8.8 (s, 1H) 8.2 (d, 1H) 8.0 (s, 1H) 7.8 (m, 2H) 7.4 (m, 4H) 7.3 (t, 2H) 7.0 (m, 3H) 3.1 (s, 3H)
From 4′-fluorobiphenyl-3-sulphonyl chloride and 1-(5-amino-2-chlorophenyl)-3-phenyl urea (Intermediate 81) using N,N-diisopropyl-N-ethylamine in NMP
LCMS (Method C) Rt 12.40 (M+H+) 496
1H NMR (400 MHz) (DMSO-d6) δ 10.5 (br s, 1H) 9.4 (s, 1H) 8.35 (s, 1H) 8.3 (s, 1H) 8.1 (s, 1H) 7.9 (d, 1H) 7.7 (m, 3H) 7.65 (t, 1H) 7.4 (d, 2H) 7.3 (m, 3H) 7.2 (t, 2H) 7.0 (t, 1H) 6.8 (d, 1H)
From 4′-fluorobiphenyl-3-sulphonyl chloride and 1-(5-aminopyridin-2-yl)-3-phenyl urea (Intermediate 66) using pyridine in DCM
LCMS (Method C) Rt 11.43 (M+H+) 463
1H NMR (400 MHz) (DMSO-d6) δ 10.2 (br s, 1H) 9.9 (s, 1H) 9.3 (s, 1H) 7.9 (m, 3H) 7.7 (m, 4H) 7.5 (m, 4H) 7.3 (m, 4H) 7.0 (t, 1H)
From 4′-fluorobiphenyl-3-sulphonyl chloride and 1-(3-amino-4-methylphenyl)-3-phenyl urea (Intermediate 63) using N,N-diisopropyl-N-ethylamine in NMP
LCMS (Method C) Rt 11.90 (M+H+) 476
1H NMR (400 MHz) (DMSO-d6) δ 9.6 (s, 1H) 8.6 (s, 1H) 8.5 (s, 1H) 7.9 (m, 2H) 7.7 (m, 4H) 7.4 (d, 2H) 7.3 (m, 5H) 7.15 (d, 1H) 7.0 (d, 1H) 6.95 (t, 1H) 1.95 (s, 3H)
From 4′-fluorobiphenyl-3-sulphonyl chloride and 1-(4-aminophenyl)-3-benzyl urea (Intermediate 26) using pyridine in DCM
LCMS (Method C) Rt 11.28 (M+H+) 476
1H NMR (400 MHz) (DMSO-d6) δ 9.9 (br s, 1H) 8.6 (s, 1H) 7.9 (m, 2H) 7.6 (m, 4H) 7.4-7.2 (m, 9H) 6.95 (d, 2H) 6.6 (t, 1H) 4.3 (d, 2H)
From 5-bromothiophene-2-sulphonyl chloride and 1-(4-methylaminophenyl)-3-phenyl urea (Intermediate 25) using pyridine in DCM.
LCMS (Method C) Rt 10.63 (M+H)+388
1H NMR (400 MHz) (DMSO-d6) δ 8.8 (s, 1H) 8.7 (s, 1H) 8.0 (d, 1H) 7.5 (m, 4H) 7.3 (m, 3H) 7.0 (m, 3H) 3.1 (s, 3H)
From 2-(4-fluorophenyl)-1-methyl-1H-imidazole-4-sulphonyl chloride (Intermediate 91) and 1-(4-methylaminophenyl)-3-phenyl urea (Intermediate 25) using pyridine in DCM
LCMS (Method C) Rt 10.54 (M+H+) 480
1H NMR (400 MHz) ((DMSO-d6) δ 8.8 (s, 1H) 8.7 (s, 1H) 7.9 (m, 3H) 7.4 (m, 6H) 7.3 (t, 2H) 7.15 (d, 2H) 6.95 (t, 1H) 3.8 (s, 3H) 3.25 (s, 3H)
From 5-(1-methyl-3-trifluoromethyl-1H-pyrazol-5-yl)thiophene-2-sulphonyl chloride and 6-amino-2,3-dihydroindole-1-carboxylic acid N-phenylamide (Intermediate 69) using pyridine in DCM
LCMS (Method C) Rt 11.82 (M+H+) 450
1H NMR (400 MHz) (DMSO-d6) δ 10.4 (br s, 1H) 8.5 (s, 1H) 7.8 (s, 1H) 7.6 (d, 1H) 7.5 (m, 3H) 7.3 (t, 2H) 7.15 (s, 1H) 7.1 (d, 1H) 7.0 (t, 1H) 6.7 (d, 1H) 4.1 (t, 2H) 4.0 (s, 3H) 3.1 (t, 2H)
From 5-(1-methyl-3-trifluoromethyl-1H-pyrazol-5-yl)thiophene-2-sulphonyl chloride and 1-(5-amino-2-methoxyphenyl)-3-phenyl urea (Intermediate 73) using pyridine in DCM
LCMS (Method C) Rt 11.74 (M+H+) 552
1H NMR (400 MHz) (DMSO-d6) δ 10.2 (br s, 1H) 9.3 (s, 1H) 8.2 (s, 1H) 8.0 (s, 1H) 7.5 (dd, 2H) 7.4 (d, 2H) 7.3 (t, 2H) 7.1 (s, 1H) 6.9 (m, 2H) 6.8 (dd, 1H) 4.0 (s, 3H) 3.8 (s, 3H)
From 2-chloro-4-methylthiazole-5-sulphonyl chloride and 1-(4-methylaminophenyl)-3-phenyl urea (Intermediate 25) using pyridine in DCM
LCMS (Method C) Rt 11.51 (M+H+) 437
1H NMR (400 MHz) (DMSO-d6) δ 8.9 (s, 1H) 8.7 (s, 1H) 7.5 (m, 4H) 7.3 (t, 2H) 7.2 (d, 2H) 7.0 (t, 1H) 3.2 (s, 3H) 2.1 (s, 3H)
From 4′-fluorobiphenyl-3-sulphonyl chloride and 1-(4-amino-3-chlorophenyl)-3-phenyl urea (Intermediate 85) using pyridine in DCM
LCMS (Method C) Rt 12.12 (M+H+) 496
1H NMR (400 MHz) (DMSO-d6) δ 10.0 (br s, 1H) 8.9 (s, 1H) 8.8 (s, 1H) 7.9 (m, 2H) 7.7 (m, 5H) 7.5 (d, 2H) 7.3 (m, 4H) 7.2 (m, 2H) 7.0 (t, 1H)
From 5-(1-methyl-3-trifluoromethyl-1H-pyrazol-5-yl)thiophene-2-sulphonyl chloride and 1-(4-amino-2-chlorophenyl)-3-phenyl urea (Intermediate 83) using pyridine in DCM
LCMS (Method C) Rt 12.10 (M+H+) 556
1H NMR (400 MHz) (DMSO-d6) δ 10.7 (br s, 1H) 9.3 (s, 1H) 8.3 (s, 1H) 8.1 (d, 1H) 7.7 (d, 1H) 7.6 (d, 1H) 7.5 (d, 2H) 7.3 (t, 2H) 7.2 (d, 2H) 7.1 (d, 1H) 7.0 (t, 1H) 4.0 (s, 3H)
From 5-chlorothiophene-2-sulphonyl chloride and 1-(4-methylaminophenyl)-3-phenyl urea (Intermediate 25) using pyridine in DCM
LCMS (Method C) Rt 11.69 (M+H+) 421
1H NMR (400 MHz) (DMSO-d6) δ 8.9 (s, 1H) 8.7 (s, 1H) 7.5 (m, 4H) 7.4 (d, 1H) 7.35 (d, 1H) 7.3 (t, 2H) 7.1 (d, 2H) 7.0 (t, 1H) 3.1 (s, 3H)
From 2,3-dihydrobenzo-1,4-dioxin-6-sulphonyl chloride and 1-(4-methylaminophenyl)-3-phenyl urea (Intermediate 25) using pyridine in DCM
LCMS (Method C) Rt 10.70 (M+H+) 440
1H NMR (400 MHz) (DMSO-d6) δ 8.8 (s, 1H) 8.7 (s, 1H) 7.4 (m, 4H) 7.3 (t, 2H) 7.0 (m, 6H) 4.35 (t, 2H) 4.3 (t, 2H) 3.1 (s, 3H)
From 3-methoxyphenylsulphonyl chloride and 1-(4-methylaminophenyl)-3-phenyl urea (Intermediate 25) using pyridine in DCM
LCMS (Method C) Rt 10.93 (M+H+) 412
1H NMR (400 MHz) (DMSO-d6) 8.8 (s, 1H) 8.7 (s, 1H) 7.5 (t, 1H) 7.4 (m, 4H) 7.3 (t, 3H) 7.1 (d, 1H) 7.0 (m, 3H) 6.9 (s, 1H) 3.8 (s, 3H) 3.1 (s, 3H)
From 4′-fluorobiphenyl-3-sulphonyl chloride and 1-(3-aminophenyl)-3-phenyl urea (Intermediate 24) using pyridine in DCM
LCMS (Method C) Rt 11.68 (M+H+) 462
1H NMR (400 MHz) (DMSO-d6) δ 10.3 (br s, 1H) 8.7 (s, 1H) 8.6 (s, 1H) 8.0 (s, 1H) 7.9 (d, 1H) 7.7 (m, 3H) 7.6 (t, 1H) 7.5 (s, 1H) 7.4 (d, 2H) 7.3 (m, 4H) 7.1 (t, 1H) 7.0 (m, 2H) 6.7 (d, 1H)
To an ice-cold solution of {N-(4′-fluorobiphenyl-3-sulphonyl)-N-[4-(3-phenylureido)-phenyl]amino}acetic acid ethyl ester (Intermediate 39, 125 mg) in THF (5 ml) was slowly added a solution of LiAlH4 in THF (1M, 250 μl). The reaction mixture was stirred for 40 minutes at 0° C. then it was quenched by addition of ethyl acetate (2 ml) followed by water (2 ml), NaOH (1M, 2 ml) and finally saturated aqueous NH4Cl (2 ml). The resultant mixture was stirred for 10 minutes then more ethyl acetate was added. The organic layer was separated, dried (Na2SO4) and filtered. The volatiles were removed by evaporation and the residue was purified by HPLC eluting with a mixture of water and acetonitrile (each containing 0.1% formic acid) from 20 to 98% acetonitrile over 25 minutes to give 4′-fluorobiphenyl-3-sulphonic acid N-(2-hydroxyethyl)-N-[4-(3-phenylureido)phenyl]amide (70 mg).
LCMS (Method C) Rt 11.18 (M+H+) 506
1H NMR (400 MHz) (DMSO-d6) δ 8.8 (br s, 1H) 8.7 (br s, 1H) 8.0 (d, 1H) 7.7 (m, 4H) 7.6 (d, 1H) 7.4 (m, 4H) 7.3 (m, 4H) 7.0 (m, 3H) 4.8 (t, 1H) 3.6 (t, 2H) 3.4 (m, 2H).
A mixture of 5-bromopyridine-3-sulphonic acid N-methyl-N-[4-(3-phenylureido)-phenyl]amide (Intermediate 6, 60 mg), 4-fluorophenyl boronic acid (36 mg), [1,1-Bis-(diphenylphosphino)ferrocene]dichloropalladium (II) (10 mg) and cesium carbonate (84 mg) in DME (1 ml) and IMS (1 ml) was heated in the microwave at 150° C. for 2 minutes. The reaction mixture was partitioned between water and ethyl acetate. The organic layer was dried (MgSO4), filtered and the volatiles were removed by evaporation. The residue was purified by HPLC eluting with a mixture of water and acetonitrile (each containing 0.1% formic acid) from 20 to 95% acetonitrile over 25 minutes to give 5-(4-fluorophenyl)pyridine-3-sulphonic acid N-methyl-N-[4-(3-phenylureido)phenyl]amide (20 mg).
LCMS (Method C) Rt 11.50 (M+H+) 477
1H NMR (400 MHz) (DMSO-d6) δ 9.2 (s, 1H) 8.8 (s, 1H) 8.7 (s, 1H) 8.6 (s, 1H) 8.0 (s, 1H) 7.8 (m, 2H) 7.4 (m, 4H) 7.3 (t, 2H) 7.2 (t, 2H) 7.1 (d, 2H) 7.0 (t, 1H) 3.2 (s, 3H)
By proceeding in a similar manner the following examples were prepared from the appropriate starting materials. The reaction may also be performed using different catalysts, bases and solvents.
From 3-pyridylboronic acid and 3-bromophenylsulphonic acid N-methylN-[4-(3-phenylureido)phenyl]amide (Intermediate 7) using [1,1′-bis(diphenylphosphino)ferrocene]-dichloropalladium (II) and aqueous Na2CO3 (2M) in acetonitrile.
LCMS (Method C) Rt 8.99 (M+H+) 459
1H NMR (400 MHz) ((DMSO-d6) δ 8.9 (m, 2H) 8.8 (s, 1H) 8.7 (d, 1H) 8.1 (t, 2H) 7.8 (m, 2H) 7.6 (m, 2H) 7.4 (m, 4H) 7.2 (t, 2H) 7.0 (d, 2H) 6.95 (t, 1H) 3.2 (s, 3H)
From 1H-pyrazole-4-boronic acid pinacol ester and 5-bromothiophene-2-sulphonic acid N-methyl-N-[4-(3-phenylureido)phenyl]amide (Example 55) using tetrakis(triphenyl-phosphine)palladium(0) and aqueous Na2CO3 (2M) in DME.
LCMS (Method C) Rt 9.78 (M+H+) 454
1H NMR (400 MHz) (DMSO-d6) δ 13.2 (br s, 1H) 8.9 (s, 1H) 8.8 (s, 1H) 8.3 (s, 1H) 7.9 (s, 1H) 7.5 (m, 4H) 7.3 (m, 4H) 7.1 (d, 2H) 6.9 (t, 1H) 3.3 (s, 3H)
From 1-methyl-1H-pyrazole-4-boronic acid and 5-bromothiophene-2-sulphonic acid N-methyl-N-[4-(3-phenylureido)phenyl]amide (Example 55) using tetrakis(triphenyl-phosphine)palladium(0) and aqueous Na2CO3 (2M) in acetonitrile.
LCMS (Method C) Rt 10.40 (M+H+) 468
1H NMR (400 MHz) (DMSO-d6) δ 8.8 (s, 1H) 8.7 (s, 1H) 8.2 (s, 1H) 7.8 (s, 1H) 7.5 (m, 4H) 7.4 (d, 1H) 7.3 (m, 3H) 7.1 (d, 2H) 7.0 (t, 1H) 3.8 (s, 3H) 3.2 (s, 3H)
From pyridine-4-boronic acid and 5-bromothiophene-2-sulphonic acid N-methyl-N-[4-(3-phenylureido)phenyl]amide (Example 55) using tetrakis(triphenylphosphine)palladium(0) and aqueous Na2CO3 (2M) in DME.
LCMS (Method C) Rt 8.63 (M+H+) 465
1H NMR (400 MHz) (DMSO-d6) δ 8.8 (s, 1H) 8.7 (s, 1H), 8.6 (d, 2H) 7.9 (d, 1H) 7.7 (d, 2H) 7.5 (d, 1H) 7.4 (m, 4H) 7.3 (t, 2H) 7.1 (d, 2H) 6.9 (t, 1H) 3.2 (s, 3H)
From pyridine-3-boronic acid and 5-bromothiophene-2-sulphonic acid N-methyl-N-[4-(3-phenylureido)phenyl]amide (Example 55) using tetrakis(triphenylphosphine)palladium(0) and aqueous Na2CO3 (2M) in DME.
LCMS (Method C) Rt 10.07 (M+H+) 465
1H NMR (400 MHz) (DMSO-d6) δ 9.0 (s, 1H) 8.8 (s, 1H) 8.7 (s, 1H) 8.6 (d, 1H) 8.2 (dd, 1H) 7.8 (d, 1H) 7.5 (m, 6H) 7.3 (t, 2H) 7.1 (d, 2H) 6.9 (t, 1H) 3.2 (s, 3H)
From 5-pyrimidyl boronic acid hydrate and 5-bromothiophene-2-sulphonic acid N-methyl-N-[4-(3-phenylureido)phenyl]amide (Example 55) using tetrakis(triphenylphosphine)-palladium(0) and aqueous Na2CO3 (2M) in DME
LCMS (Method C) Rt 10.05 (M+H+) 466
1H NMR (400 MHz) (DMSO-d6) δ 9.3 (s, 1H) 9.2 (s, 2H) 9.15 (s, 1H) 9.1 (s, 1H) 7.9 (d, 1H) 7.55 (d, 1H) 7.5 (d, 4H) 7.3 (t, 2H) 7.1 (d, 2H) 6.9 (t, 1H) 3.2 (s, 3H)
From cyclohexen-1-yl boronic acid and 5-bromothiophene-2-sulphonic acid N-methyl-N-[4-(3-phenylureido)phenyl]amide (Example 55) using tetrakis(triphenylphosphine)palladium(0) and aqueous Na2CO3 (2M) in DME
LCMS (Method C) Rt 13.41 (M+H+) 468
1H NMR (400 MHz) (DMSO-d6) δ 9.5 (s, 1H) 9.4 (s, 1H) 7.5 (m, 4H) 7.3 (m, 3H) 7.1 (d, 1H) 7.0 (d, 2H) 7.0 (t, 1H) 6.3 (m, 1H) 3.1 (s, 3H) 2.3 (m, 2H) 2.1 (m, 2H) 1.7 (m, 2H) 1.6 (m, 2H)
From 1-methyl-1H-pyrazole-5-boronic acid pinacol ester and 5-bromothiophene-2-sulphonic acid N-methyl-N-[4-(3-phenylureido)phenyl]amide (Example 55) using tetrakis(triphenylphosphine)palladium(0) and aqueous Na2CO3 (2M) in DME
LCMS (Method C) Rt 10.65 (M+H+) 468
1H NMR (400 MHz) (DMSO-d6) δ 8.8 (s, 1H) 8.7 (s, 1H) 7.5 (m, 3H) 7.4 (m, 4H) 7.3 (t, 2H) 7.1 (d, 2H) 7.0 (t, 1H) 6.6 (s, 1H) 4.0 (s, 3H) 3.2 (s, 3H)
From 3,5-dimethylisoxazol-4-boronic acid and 5-bromothiophene-2-sulphonic acid N-methyl-N-[4-(3-phenylureido)phenyl]amide (Example 55) using tetrakis(triphenylphosphine)palladium(0) and aqueous Na2CO3 (2M) in DME
LCMS (Method C) Rt 11.43 (M+H+) 483
1H NMR (400 MHz) (DMSO-d6) δ 8.8 (s, 1H) 8.7 (s, 1H) 7.6 (d, 1H) 7.5 (m, 4H) 7.4 (d, 1H) 7.3 (t, 2H) 7.1 (d, 2H) 7.0 (t, 1H) 3.2 (s, 3H) 2.5 (s, 3H) 2.3 (s, 3H)
From 1-methyl-3-trifluoromethyl-1H-pyrazol-5-boronic acid and 3-bromophenylsulphonic acid N-methyl-N-[4-(3-phenylureido)phenyl]amide (Intermediate 7) using tetrakis(triphenylphosphine)palladium(0) and aqueous Na2CO3 (2M) in DME
LCMS (Method C) Rt 12.12 (M+H+) 530
1H NMR (400 MHz) (DMSO-d6) δ 8.8 (s, 1H) 8.7 (s, 1H) 8.0 (d, 1H) 7.8 (t, 1H) 7.6 (m, 2H) 7.4 (m, 4H) 7.3 (m, 2H) 7.1 (d, 2H) 7.0 (m, 2H) 3.8 (s, 3H) 3.1 (s, 3H)
From 2-methoxypyridyl-5-boronic acid and 5-bromothiophene-2-sulphonic acid N-methyl-N-[4-(3-phenylureido)phenyl]amide (Example 55) using tetrakis(triphenylphosphine)palladium(0) and aqueous Na2CO3 (2M) in DME
LCMS (Method C) Rt 11.94 (M+H+) 495
1H NMR (400 MHz) (DMSO-d6) δ 8.8 (s, 1H) 8.7 (s, 1H) 8.6 (d, 1H) 8.1 (dd, 1H) 7.6 (d, 1H) 7.5 (m, 5H) 7.3 (t, 2H) 7.1 (d, 2H) 6.9 (m, 2H) 3.9 (s, 3H) 3.1 (s, 3H)
From 1-methyl-1-H-pyrazole-5-boronic acid pinacol ester and 3-bromophenylsulphonic acid N-methyl-N-[4-(3-phenylureido)phenyl]amide (Intermediate 7) using tetrakis(triphenylphosphine)palladium(0) and aqueous Na2CO3 (2M) in DME
LCMS (Method C) Rt 10.38 (M+H+) 462
1H NMR (400 MHz) ((DMSO-d6) δ 8.9 (s, 1H) 8.8 (s, 1H) 7.9 (d, 1H) 7.7 (t, 1H) 7.6 (d, 1H) 7.5 (m, 6H) 7.3 (m, 2H) 7.0 (m, 3H) 6.4 (s, 1H) 3.8 (s, 3H) 3.2 (s, 3H)
From 1-methyl-1H-pyrazole-5-boronic acid pinacol ester and 5-chloro-4-bromothiophene-2-sulphonic acid N-methyl-N-[4-(3-phenylureido)phenyl]amide (Intermediate 15) using tetrakis(triphenylphosphine)palladium(0) and aqueous Na2CO3 (2M) in DME.
LCMS (Method C) Rt 11.32 (M+H+) 501
1H NMR (400 MHz) (DMSO-d6) δ 8.8 (br s, 1H) 8.7 (br s, 1H) 7.6 (s, 1H) 7.5 (d, 1H) 7.45-7.4 (m, 4H) 7.2 (t, 2H) 7.1 (d, 2H) 6.9 (t, 1H) 6.5 (d, 1H) 3.7 (s, 3H) 3.2 (s, 3H).
From cyclopropylboronic acid and 5-bromothiophene-2-sulphonic acid N-methyl-N-[4-(3-phenylureido)phenyl]amide (Example 55) using tetrakis(triphenylphosphine)palladium(0) and aqueous Na2CO3 (2M) in DME.
LCMS (Method C) Rt 11.77 (M+H+) 428
1H NMR (400 MHz) (DMSO-d6) δ 8.8 (br s, 1H) 8.7 (br s, 1H) 7.5-7.4 (m, 4H) 7.3 (t, 2H) 7.25 (d, 1H) 7.1 (d, 2H) 7.0 (t, 1H) 6.95 (d, 1H) 3.2 (s, 3H) 2.3-2.2 (m, 1H) 1.15-1.10 (m, 2H) 0.8-0.75 (m, 2H)
From 4-methylpyridine-3-boronic acid and 5-bromothiophene-2-sulphonic acid N-methyl-N-[4-(3-phenylureido)phenyl]amide (Example 55) using [1,1-Bis(diphenylphosphino)ferrocene]dichloropalladium (II) and aqueous Na2CO3 (2M) in acetonitrile.
LCMS (Method C) Rt 9.28 (M+H+) 479
1H NMR (400 MHz) (DMSO-d6) δ 8.9 (br s, 1H) 8.8 (br s, 1H) 8.5 (s, 1H) 8.4 (d, 1H) 7.5 (d, 1H) 7.45-7.4 (m, 5H) 7.35 (d, 1H) 7.2 (t, 2H) 7.0 (d, 2H) 6.9 (t, 1H) 3.2 (s, 3H) 2.4 (s, 3H).
From 2-methoxypyridine-3-boronic acid and 5-bromothiophene-2-sulphonic acid N-methyl-N-[4-(3-phenylureido)phenyl]amide (Example 55) using [1,1′-Bis(diphenylphosphino)ferrocene]dichloropalladium (II) and aqueous Na2CO3 (2M) in acetonitrile.
LCMS (Method C) Rt 12.00 (M+H+) 495
1H NMR (400 MHz) (DMSO-d6) δ 8.9 (br s, 1H) 8.8 (br s, 1H) 8.3 (dd, 1H) 8.2 (dd, 1H) 7.8 (d, 1H) 7.45-7.35 (m, 5H) 7.2 (t, 2H) 7.1 (dd, 1H) 7.0 (d, 2H) 6.9 (t, 1H) 4.0 (s, 3H) 3.1 (s, 3H).
From 2-methoxypyrimidine-5-boronic acid and 5-bromothiophene-2-sulphonic acid N-methyl-N-[4-(3-phenylureido)phenyl]amide (Example 55) using [1,1-Bis(diphenylphosphino)ferrocene]dichloropalladium (II) and aqueous Na2CO3 (2M) in acetonitrile.
LCMS (Method C) Rt 10.95 (M+H+) 496
1H NMR (400 MHz) (DMSO-d6) δ 9.0 (s, 2H) 8.8 (br s, 1H) 8.7 (br s, 1H) 7.7 (d, 1H) 7.5 (d, 1H) 7.45 (m, 4H) 7.3 (t, 2H) 7.1 (d, 2H) 7.0 (t, 1H) 4.0 (s, 3H) 3.2 (s, 3H).
From 2-(dimethylamino)pyridine-5-boronic acid and 5-bromothiophene-2-sulphonic acid N-methyl-N-[4-(3-phenylureido)phenyl]amide (Example 55) using [1,1-Bis(diphenylphosphino)ferrocene]dichloropalladium (II) and aqueous Na2CO3 (2M) in acetonitrile.
LCMS (Method C) Rt 8.87 (M+H+) 508
1H NMR (400 MHz) (DMSO-d6) δ 8.7 (br s, 1H) 8.6 (br s, 1H) 8.4 (d, 1H) 7.8 (dd, 1H) 7.4 (m, 5H) 7.3 (d, 1H) 7.2 (t, 2H) 7.05 (d, 2H) 6.9 (t, 1H) 6.7 (d, 1H) 3.1 (s, 3H) 3.0 (s, 6H).
A mixture of 5-bromothiophene-2-sulphonic acid N-methyl-N-[4-(3-phenylureido)phenyl]amide (Example 55, 93 mg), 2-tributylstannyl oxazole (255 μl) and tetrakis(triphenylphosphine)palladium(0), (23 mg) in DME (3 ml) was heated in the microwave at 150° C. for 45 minutes. The reaction mixture was diluted with water and extracted with ethyl acetate. The organic layer was washed with water, dried (MgSO4), filtered and the volatiles were removed by evaporation. The residue was purified by HPLC eluting with a mixture of water and acetonitrile (each containing 0.1% formic acid) from 50 to 70% acetonitrile over 25 minutes to give 5-(oxazol-2-yl)thiophene-2-sulphonic acid N-methyl-N-[4-(3-phenylureido)phenyl]amide (6 mg).
LCMS (Method C) Rt 10.86 (M+H+) 455
1H NMR (400 MHz) (DMSO-d6) δ 8.8 (s, 1H) 8.7 (s, 1H) 8.3 (s, 1H) 7.8 (d, 1H) 7.5 (d, 1H) 7.45 (m, 5H) 7.3 (t, 2H) 7.1 (d, 2H) 6.95 (t, 1H) 3.2 (s, 3H)
By proceeding in a similar manner the following compounds were prepared from the appropriate starting materials:
From 1-methyl-5-tributylstannyl-1H-imidazole and 5-bromothiophene-2-sulphonic acid N-methyl-N-[4-(3-phenylureido)phenyl]amide (Example 55)
LCMS (Method C) Rt 7.33 (M+H+) 468
1H NMR (400 MHz) (DMSO-d6) δ 8.9 (br s, 1H) 8.8 (br s, 1H) 7.8 (s, 1H) 7.5 (m, 5H) 7.4 (d, 1H) 7.3 (m, 3H) 7.1 (d, 2H) 7.0 (t, 1H) 3.8 (s, 3H) 3.2 (s, 3H)
From 3-methyl-2-(tributylstannyl)pyridine and 5-bromothiophene-2-sulphonic acid N-methyl-N-[4-(3-phenylureido)phenyl]amide (Example 55)
LCMS (Method C) Rt 11.73 (M+H+) 479
1H NMR (400 MHz) (DMSO-d6) δ 8.8 (br s, 1H) 8.7 (br s, 1H) 8.4 (dd, 1H) 7.8 (dd, 1H) 7.65 (d, 1H) 7.45 (m, 5H) 7.3 (dd, 1H) 7.25 (t, 2H) 7.1 (t, 2H) 7.0 (m, 1H) 3.2 (s, 3H) 2.6 (s, 3H).
A mixture of 5-bromothiophene-2-sulphonic acid N-methyl-N-[4-(3-phenylureido)-phenyl]amide (Example 55, 140 mg), pyrazole (31 mg), cesium carbonate (196 mg), copper (I) oxide (2 mg) and salicylaldoxime (8 mg) in acetonitrile (1 ml) was heated in a sealed vial at 85° C. for 24 hours. The reaction mixture was diluted with water and extracted with ethyl acetate. The organic layer was washed with water, dried (MgSO4), filtered and the volatiles were removed by evaporation. The residue was purified by chromatography on a 5 g silica cartridge eluting with a mixture of ethyl acetate and dichloromethane (1:39 increasing to 1:19). The resultant solid was recrystallised from a mixture of ethyl acetate and pentane to give 5-(1H-pyrazol-1-yl)thiophene-2-sulphonic acid N-methyl-N-[4-(3-phenylureido)-phenyl]amide (47 mg).
LCMS (Method C) Rt 11.11 (M+H+) 454
1H NMR (400 MHz) (DMSO-d6) δ 8.8 (br s, 1H) 8.7 (br s, 1H) 8.6 (d, 1H) 7.8 (d, 1H) 7.5-7.4 (m, 6H) 7.3 (t, 2H) 7.1 (d, 2H) 7.0 (t, 1H) 6.6 (t, 1H) 3.2 (s, 3H).
By proceeding in a similar manner the following compound was prepared from the appropriate starting materials:
From 5-bromothiophene-2-sulphonic acid N-methyl-N-[4-(3-phenylureido)-phenyl]amide (Example 55) and imidazole.
LCMS (Method C) Rt 8.33 (M+H+) 453
1H NMR (400 MHz) (DMSO-d6) δ 9.0 (br s, 1H) 8.9 (br s, 1H) 8.2 (s, 1H) 7.7 (t, 1H) 7.45-7.4 (m, 6H) 7.2 (t, 2H) 7.1-7.05 (m, 3H) 6.9 (t, 1H) 3.15 (s, 3H).
A solution of 4′-fluorobiphenyl-3-sulphonic acid N-(3-benzyloxypropyl)-N-[4-(3-phenylureido)phenyl]amide (Intermediate 43, 151 mg), palladium on carbon (10%, 29 mg) and acetic acid (128 □l) in IMS (2.5 ml) was hydrogenated under a balloon of hydrogen at atmospheric pressure overnight. The catalyst was removed by filtration through Celite under nitrogen and the filtrate was concentrated to dryness. The residue was purified by HPLC eluting with a mixture of water and acetonitrile (each containing 0.1% formic acid) from 20 to 95% acetonitrile over 25 minutes to give 4′-fluorobiphenyl-3-sulphonic acid N-(3-hydroxypropyl)-N-[4-(3-phenylureido)phenyl]amide (35 mg).
LCMS (Method C) Rt 11.28 (M+H+) 520
1H NMR (400 MHz) ((DMSO-d6) δ 8.8 (s, 1H) 8.7 (s, 1H), 8.0 (d, 1H) 7.7 (m, 4H) 7.6 (d, 1H) 7.5 (m, 4H) 7.3 (m, 4H) 7.0 (m, 3H) 4.4 (t, 1H) 3.6 (t, 2H) 3.4 (m, 2H) 1.5 (m, 2H)
1-(4-Fluorophenyl)-3,5-dimethyl-1H-pyrazole-4-sulphonic acid [4-(3-phenylureido)phenyl]-amide (Example 24, 202 mg) was dissolved in THF (6.0 ml). Triphenylphosphine (221 mg) was added followed by 3-benzyloxy-1-propanol (133 μl). The mixture was cooled to 0° C. and diethyl azodicarboxylate (132 μl) was slowly added and the reaction mixture was stirred overnight at room temperature. The volatiles were removed by evaporation and the residue was purified by chromatography using the Biotage system on a 10 g silica cartridge eluting with a mixture of ethyl acetate and cyclohexane (1:19 increasing to 3:7) to give 1-(4-fluorophenyl)-3,5-dimethyl-1H-pyrazole-4-sulphonic acid N-(3-benzyloxypropyl)-N-[4-(3-phenylureido)phenyl]amide (339 mg).
LCMS (Method A) Rt 4.23 (M+H+) 628
The deprotection step was carried out in a similar manner to that used for Example 91.
LCMS (Method C) Rt 10.20 (M+H+) 538
1H NMR (400 MHz) ((DMSO-d6) δ 8.8 (s, 1H) 8.7 (s, 1H) 7.6 (m, 2H) 7.4 (m, 6H) 7.3 (t, 2H) 7.1 (d, 2H) 7.0 (t, 1H) 4.4 (t, 1H) 3.6 (t, 2H) 3.4 (m, 2H) 2.1 (s, 3H) 2.0 (s, 3H) 1.5 (m, 2H)
The alkylation was performed in a similar manner to that used in Example 92, Step 1 starting from 5-(pyridin-2-yl)thiophene-2-sulphonic acid [4-(3-phenylureido)phenyl]amide (Example 32) and 3-tert-butoxy-1-propanol
LCMS (Method A) Rt 4.16 (M+H+) 565
To an ice-cooled solution of 5-(pyridin-2-yl)thiophene-2-sulphonic acid N-(3-tert-butoxypropyl)-N-[4-(3-phenylureido)phenyl]amide (from step 1, 425 mg) in DCM (9 ml) and MeOH (1 ml) was added TFA (5 ml). The resultant mixture was stirred at room temperature overnight. The volatiles were removed by evaporation and the residue was dissolved in THF (10 ml) and treated with NaOH (1M, 5 ml). The resultant mixture was stirred at room temperature for 1 hour. The volatiles were removed by evaporation and the residue was purified by HPLC eluting with a mixture of water and acetonitrile (each containing 0.1% formic acid) from 50 to 70% acetonitrile over 20 minutes to give 5-(pyridin-2-yl)thiophene-2-sulphonic acid N-(3-hydroxypropyl)-N-[4-(3-phenylureido)phenyl]amide (325 mg).
LCMS (Method C) Rt 10.03 (M+H+) 509
1H NMR (400 MHz) ((DMSO-d6) δ 8.8 (s, 1H) 8.7 (s, 1H) 8.6 (d, 1H) 8.1 (d, 1H) 7.9 (m, 2H) 7.5 (m, 6H) 7.3 (t, 2H) 7.0 (d, 2H) 6.9 (t, 1H) 4.5 (t, 1H) 3.7 (t, 2H) 3.4 (dt, 2H) 1.3 (t, 2H)
A solution of 4′-fluorobiphenyl-3-sulphonic acid N-(3-tert-butoxypropyl)-N-[2-methoxy-4-(3-phenylureido)phenyl]amide (Intermediate 38, 110 mg) in DCM (2 ml) was treated with TFA (1 ml). The resultant mixture was stirred at room temperature for 2 hours, then diluted with DCM and washed with water. The organic layer was dried (MgSO4), filtered and the volatiles were removed by evaporation. The residue was dissolved in MeOH (5 ml) and treated with HCl (1M, 1.7 ml). The resultant mixture was stirred at room temperature for 2 hours, concentrated and partitioned between water and ethyl acetate. The aqueous layer was further extracted with ethyl acetate and the combined organic layers were washed with water, dried (MgSO4) and filtered. The volatiles were removed by evaporation to give 4′-fluorobiphenyl-3-sulphonic acid N-(3-hydroxypropyl)-N-[2-methoxy-4-(3-phenylureido)-phenyl]amide as an off-white solid (74 mg).
LCMS (Method C) Rt 11.35 (M+H+) 550
1H NMR (400 MHz) ((DMSO-d6) δ 8.8 (s, 1H) 8.7 (s, 1H) 7.9 (dd, 1H) 7.7 (m, 5H) 7.5 (d, 2H) 7.3 (m, 5H) 7.1 (d, 1H) 7.0 (t, 1H) 6.9 (dd, 1H) 4.2 (br s, 1H) 3.6 (br s, 2H) 3.4 (t, 2H) 3.3 (s, 3H) 1.5 (m, 2H)
The alkylation was performed in a similar manner to that used for Example 92, Step1 starting from 1-(4-fluorophenyl)-1H-pyrazole-4-sulphonic acid [4-(3-phenylureido)phenyl]amide (Intermediate 8) and 3-tert-butoxy-1-propanol
LCMS (Method A) Rt 4.15 (M+H+) 566
The deprotection step was carried out in a similar manner to that used for Example 94
LCMS (Method C) Rt 10.33 (M+H+) 510
1H NMR (400 MHz) ((DMSO-d6) δ 9.0 (s, 1H) 8.8 (s, 1H) 8.7 (s, 1H) 8.0 (m, 2H), 7.9 (s, 1H) 7.4 (m, 6H) 7.3 (t, 2H) 7.1 (d, 2H) 7.0 (t, 1H) 4.4 (t, 1H) 3.6 (t, 2H) 3.4 (m, 2H) 1.5 (m, 2H)
The alkylation step was carried out in a similar manner to that used for Example 92, Step1 starting from 5-(1-methyl-3-trifluoromethyl-1H-pyrazol-5-yl)thiophene-2-sulphonic acid [4-(3-phenylureido)phenyl]amide (Example 33) and 3-tert-butoxypropan-1-ol
LCMS (Method B) Rt 4.48 (M+H+) 636
The deprotection was carried out in a similar manner to that used for Example 94, Step 2.
LCMS (Method C) Rt 11.14 (M+H+) 580
1H NMR (400 MHz) ((DMSO-d6) δ 8.9 (s, 1H) 8.7 (s, 1H) 7.7 (d, 1H) 7.6 (d, 1H) 7.5 (m, 4H) 7.3 (t, 2H) 7.2 (s, 1H) 7.1 (d, 2H) 7.0 (t, 1H) 4.5 (t, 1H) 4.1 (s, 3H) 3.6 (t, 2H) 3.4 (m, 2H) 1.5 (m, 2H)
To a suspension of 5-[1-(tetrahydropyran-2-yl)-1H-pyrazol-5-yl]-thiophene-2-sulphonic acid N-methyl-N-[4-(3-phenylureido)phenyl]amide (Intermediate 33, 62 mg) in MeOH (1 ml) was added a solution of HCl in MeOH (1.25 M, 2.0 ml). The mixture was stirred for 1 h, then diluted with water and treated with saturated aqueous NaHCO3. The resultant mixture was extracted with ethyl acetate and the organic layer was washed with water, dried (MgSO4), filtered and the volatiles were removed by evaporation. The residue was purified by HPLC eluting with a mixture of water and acetonitrile (each containing 0.1% formic acid) from 50 to 70% acetonitrile over 20 minutes to give 5-(1H-pyrazol-5-yl)thiophene-2-sulphonic acid N-methyl-N-[4-(3-phenylureido)phenyl]amide (24 mg).
LCMS (Method C) Rt 10.10 (M+H+) 454
1H NMR (400 MHz) ((DMSO-d6) δ 13.1 (br s, 1H) 8.8 (s, 1H) 8.7 (s, 1H) 7.8 (s, 1H) 7.4 (m, 6H) 7.3 (t, 2H) 7.1 (d, 2H) 7.0 (t, 1H) 6.8 (s, 1H) 3.2 (s, 3H)
To an ice-cooled solution of 4′-fluorobiphenyl-3-sulphonic acid N-(3-hydroxypropyl)-N-[4-(3-phenylureido)phenyl]amide (Example 91, 100 mg) and triethylamine (30 μl) in DCM (3 ml) was added methanesulphonyl chloride (17 μl). The mixture was stirred at room temperature for 3 hours and then further methanesulphonyl chloride was added (10 μl). After a further 1 hour, the reaction mixture was poured into a mixture of ice and water, and extracted with DCM. The organic layer was dried (Na2SO4), filtered and the volatiles were removed by evaporation to give 4′-fluorobiphenyl-3-sulphonic acid N-(3-methanesulphonyloxypropyl)-N-[4-(3-phenylureido)phenyl]amide (84 mg).
LCMS (Method A) Rt. 3.95 (M+H+) 598
A mixture of 4′-fluorobiphenyl-3-sulphonic acid N-(3-methanesulphonyloxypropyl)-N-[4-(3-phenylureido)phenyl]amide (from Step 1, 68 mg) and dimethylamine (40% in water, 86 μl) in 1,4-dioxane (230 μl) was heated in the microwave at 125° C. for 5 minutes. The resultant mixture was concentrated and the residue was purified by HPLC eluting with a mixture of water and acetonitrile (each containing 0.1% formic acid) from 20 to 98% acetonitrile over 25 minutes to give 4′-fluorobiphenyl-3-sulphonic acid N-(3-dimethylaminopropyl)-N-[4-(3-phenylureido)phenyl]amide (12 mg).
LCMS (Method C) Rt 8.41 (M+H+) 547
1H NMR (400 MHz) ((DMSO-d6) δ 9.2 (s, 1H) 9.1 (s, 1H) 8.2 (s, 1H) 8.0 (d, 1H) 7.7 (m, 4H) 7.6 (d, 1H) 7.5 (m, 3H) 7.3 (m, 4H) 7.0 (m, 3H) 3.6 (t, 2H) 2.3 (t, 2H) 2.1 (s, 6H) 1.5 (m, 2H)
A mixture of 4′-fluorobiphenyl-3-sulphonic acid N-[(2,2-dimethyl-1,3-dioxolan-4-yl)methyl]-N-[4-(3-phenylureido)phenyl]amide (Intermediate 45, 57 mg) in acetone (2 ml) containing HCl (1M, 1 ml) was heated at reflux for 4 hours. The mixture was concentrated and the residue was purified by HPLC eluting with a mixture of water and acetonitrile (each containing 0.1% formic acid) from 20 to 98% acetonitrile over 30 minutes to give 4′-fluorobiphenyl-3-sulphonic acid N-(2,3-dihydroxypropyl)-N-[4-(3-phenylureido)phenyl]-amide (23 mg).
LCMS (Method C) Rt 10.40 (M+H+) 536
1H NMR (400 MHz) ((DMSO-d6) δ 8.9 (s, 1H) 8.8 (s, 1H) 8.0 (d, 1H) 7.7 (m, 4H) 7.6 (d, 1H) 7.4 (m, 4H) 7.3 (m, 4H) 7.0 (m, 3H) 4.8 (d, 1H) 4.5 (t, 1H) 3.5 (m, 2H) 3.4-3.2 (m, 3H)
5-(Cyclohexen-1-yl)thiopheny-2-sulphonic acid N-methyl-N-[4-(3-phenylureido)phenyl]-amide (Example 74, 32 mg) was dissolved in a mixture of ethyl acetate (2 ml) and THF (1 ml) and treated with palladium on carbon (10%, 65 mg). The reaction mixture was hydrogenated under a balloon of hydrogen at atmospheric pressure for 4 hours. The catalyst was removed by filtration through Celite under nitrogen and the filtrate was concentrated to dryness to give 5-cyclohexylthiophene-2-sulphonic acid N-methyl-N-[4-(3-phenylureido)phenyl]amide (20 mg) as a white solid.
LCMS (Method C) Rt 13.57 (M+H+) 470
1H NMR (400 MHz) (DMSO-d6) δ 9.2 (s, 1H) 9.1 (s, 1H) 7.5 (m, 4H) 7.3 (m, 3H) 7.0 (m, 4H) 3.2 (s, 3H) 2.9 (m, 1H) 2.0 (m, 2H) 1.75 (m, 2H) 1.6 (m, 1H) 1.4 (m, 4H) 1.3 (m, 1H)
5-(1-Boc-3,6-dihydro-2H-pyridin-4-yl)thiophene-2-sulphonic acid N-methyl-N-[4-(3-phenyl-ureido)phenyl]amide (Intermediate 35, 45 mg) was dissolved in dichloromethane (1 ml) and treated with TFA (1 ml). The mixture was stirred at room temperature for 1 hour then the volatiles were removed by evaporation. The residue was purified by passing through an SCX-2 column eluting with DCM followed by a mixture of DCM and methanol (1:1) and finally a mixture of DCM and 2M ammonia in methanol (1:1). After evaporation of the volatiles, the residue was purified by HPLC eluting with a mixture of water and acetonitrile (each containing 0.1% formic acid) from 20 to 98% acetonitrile to give 5-(1,2,3,6-tetrahydropyridin-4-yl)thiophene-2-sulphonic acid N-methyl-N-[4-(3-phenylureido)phenyl]-amide (20 mg).
LCMS (Method C) Rt 7.20 (M+H+) 469
1H NMR (400 MHz) (DMSO-d6) δ 9.5 (s, 1H) 9.4 (s, 1H) 7.5-7.4 (m, 4H) 7.3 (d, 1H) 7.2 (t, 2H) 7.16 (d, 1H) 6.3 (br s, 1H) 3.9-3.2 (broad signal) 3.1 (s, 3H) 3.0 (broad signal), 2.4 (broad signal).
Prepared by proceeding in a similar manner to that used for Example 100 starting from 5-(1-Boc-3,6-dihydro-2H-pyridin-4-yl)thiophene-2-sulphonic acid N-methyl-N-[4-(3-phenyl-ureido)phenyl]amide (Intermediate 35).
LCMS (Method B) Rt 4.20 (M+H+) 571
The deprotection was carried out in a similar manner to that used for Example 101.
LCMS (Method C) Rt 7.15 (M+H+) 471
1H NMR (400 MHz) (DMSO-d6) 8.85 (br s, 1H) 8.75 (br s, 1H) 7.45 (t, 4H) 7.3 (m, 3H) 7.0 (m, 3H) 6.95 (t, 1H) 3.15 (s, 3H) 3.0 (m, 3H) 2.55 (dt, 2H) 1.85 (m, 2H) 1.45 (dq, 2H).
A mixture of 5-(piperidin-4-yl)thiophene-2-sulphonic acid N-methylN-[4-(3-phenylureido)-phenyl]amide (from Step 2, 235 mg), formaldehyde (60 μl), acetic acid (60 μl), and sodium triacetoxyborohydride (60 mg) in dichloromethane (5 ml) was stirred at room temperature under nitrogen for 18 hours. The mixture was quenched with water and extracted with DCM and then with ethyl acetate. The combined organic layers were dried (Na2SO4), filtered and the volatiles were removed by evaporation. The residue was purified by chromatography using a 5 g silica cartridge eluting with DCM followed by a mixture of DCM and 2M ammonia in methanol (9:1) to give 5-(1-methylpiperidin-4-yl)thiophene-2-sulphonic acid N-methyl-N-[4-(3-phenylureido)phenyl]amide as a white solid (32 mg)
LCMS (Method C) Rt 7.20 (M+H+) 485
1H NMR (400 MHz) (DMSO-d6) δ 8.8 (br s, 1H) 8.7 (br s, 1H) 7.45 (m, 4H) 7.3 (m, 3H) 7.0 (m, 3H) 6.95 (t, 1H) 3.1 (s, 3H) 2.8 (m, 3H) 2.1 (s, 3H) 1.95 (m, 4H) 1.6 (m, 2H).
5-(4,4,4-Trifluoro-1,3-dioxobutyl)thiophene-2-sulphonic acid N-methyl-N-[4-(3-phenyl-ureido)phenyl]amide (Intermediate 9, 34 mg) was dissolved in IMS (1 ml) and hydrazine hydrate (30 μl) was added. The mixture was stirred at room temperature for 1 hour then heated at 70° C. for 3 hours and allowed to stand at room temperature overnight. The reaction was repeated using 5-(4,4,4-trifluoro-1,3-dioxobutyl)thiophene-2-sulphonic acid N-methyl-N-[4-(3-phenylureido)phenyl]amide (Intermediate 9, 96 mg), IMS (3 ml) and hydrazine hydrate (83 μl). The two crude reaction mixtures were combined and poured into water and extracted with ethyl acetate. The organic layer was washed with water, dried (MgSO4), filtered and the volatiles were removed by evaporation. The residue was purified by HPLC eluting with a mixture of water and acetonitrile (each containing 0.1% formic acid) from 50 to 70% acetonitrile over 20 minutes to give 5-(3-trifluoromethyl-1H-pyrazol-5-yl)thiophene-2-sulphonic acid N-methyl-N-[4-(3-phenylureido)phenyl]amide (11 mg).
LCMS (Method C) Rt 11.62 (M+H+) 522
1H NMR (400 MHz) ((DMSO-d6) δ 14.5 (br s, 1H) 8.9 (s, 1H) 8.8 (s, 1H) 7.65 (d, 1H) 7.55 (d, 1H) 7.5 (m, 4H) 7.3 (m, 3H) 7.1 (d, 2H) 7.0 (t, 1H) 3.2 (s, 3H)
A mixture of 4-fluorophenylboronic acid (160 mg), copper acetate (65 mg) and 1H-imidazole-4-sulphonic acid N-methyl-N-[4-(3-phenylureido)phenyl]amide (Intermediate 16, 98 mg) in pyridine (2 ml) containing 4 A Molecular Sieves was stirred at room temperature for 18 hours and then heated at 40° C. further 18 hours. The mixture was filtered through Celite and the filtrate was diluted with water and extracted with DCM. The organic layer was dried (Na2SO4), filtered and the volatiles were removed by evaporation. The residue was purified by chromatography using a 5 g silica cartridge eluting with DCM and then a mixture of ethyl acetate and DCM (1:9 to 1:4). The resultant product was further purified by HPLC eluting with a mixture of water and acetonitrile (each containing 0.1% formic acid) from 40 to 60% acetonitrile over 20 minutes to give 1-(4-fluorophenyl)-1H-imidazole-4-sulphonic acid N-methyl-N-[4-(3-phenylureido)phenyl]amide as a white solid (28 mg).
LCMS (Method C) Rt 10.49 (M+H+) 466
1H NMR (400 MHz) (DMSO-d6) δ 8.7 (br s, 1H) 8.6 (br s, 1H) 8.4 (d, 1H) 8.2 (d, 1H) 7.8-7.7 (m, 2H) 7.4-7.3 (m, 6H) 7.2 (m, 2H) 7.1 (d, 2H) 6.9 (t, 1H) 3.2 (s, 3H).
By proceeding in a similar manner the following compounds were prepared from the appropriate starting materials:
From phenylboronic acid and 1H-pyrazole-4-sulphonic acid N-methyl-N-[4-(3-phenylureido)-phenyl]amide (Intermediate 17)
LCMS (Method C) Rt 11.36 (M+H+) 448
1H NMR (400 MHz) (DMSO-d6) δ 9.0 (s, 1H) 8.8 (s, 1H) 8.7 (s, 1H) 7.9 (d, 2H) 7.8 (s, 1H) 7.5 (t, 2H) 7.4 (m, 5H) 7.3 (t, 2H) 7.2 (d, 2H) 6.9 (t, 1H) 3.1 (s, 3H).
From 1-methyl-1-H-pyrazole-4-boronic acid and 1H-pyrazole-4-sulphonic acid N-methyl-N-[4-(3-phenyl ureido)phenyl]amide (Intermediate 17)
LCMS (Method C) Rt 9.51 (M+H+) 452
1H NMR (400 MHz) (DMSO-d6) δ 9.0 (br s, 1H) 8.9 (br s, 1H) 8.7 (s, 1H) 8.3 (s, 1H) 8.0 (s, 1H) 7.8 (s, 1H) 7.5 (t, 4H) 7.3 (t, 2H) 7.1 (d, 2H) 7.0 (t, 1H) 3.9 (s, 3H) 3.2 (s, 3H).
From 2-methoxy-5-pyridineboronic acid and 1H-pyrazole-4-sulphonic acid N-methyl-N-[4-(3-phenylureido)phenyl]amide (Intermediate 17)
LCMS (Method C) Rt 10.98 (M+H+) 479
1H NMR (400 MHz) (DMSO-d6) 9.0 (s, 1H) 8.8 (s, 1H) 8.7 (d, 1H) 8.65 (s, 1H) 8.2 (dd, 1H) 7.8 (s, 1H) 7.45 (m, 4H) 7.3 (m, 2H) 7.1 (m, 2H) 7.0 (m, 2H) 3.9 (s, 3H) 3.2 (s, 3H).
| Number | Date | Country | Kind |
|---|---|---|---|
| 20095678 | Jun 2009 | FI | national |
| Filing Document | Filing Date | Country | Kind | 371c Date |
|---|---|---|---|---|
| PCT/FI10/50495 | 6/14/2010 | WO | 00 | 2/15/2012 |
