HK1117153A

HK1117153A - Substituted pyrroles, compositions containing same, method for making same and use thereof

Info

Publication number: HK1117153A
Application number: HK08108016.5A
Authority: HK
Inventors: Baptiste Ronan; Michel Tabart; Catherine Souaille; Fabrice Viviani; Eric Bacque; Jean-Philippe Lettalec; Pascal Desmazeau
Original assignee: Aventis Pharma Sa
Priority date: 2005-02-10
Filing date: 2006-02-08
Publication date: 2009-01-09

Description

substituted pyrroles, compositions containing them, processes for their preparation and their use

The present invention relates to particularly novel chemical compounds, in particular novel substituted pyrroles, compositions containing them and their use as pharmaceuticals.

More particularly, the invention relates to novel specific pyrroles having anti-cancer activity by modulating protein activity, particularly kinase activity.

To date, the major problems with most commercial compounds used in chemotherapy have been side effects and patient tolerance. These side effects can be limited as long as the drug used selectively acts on cancer cells (except healthy cells). One solution to limit the adverse effects of chemotherapy may therefore consist in using drugs that act on metabolic pathways or on the constituent elements of these pathways, which are mostly expressed in cancer cells and not or less expressed in healthy cells.

Protein kinases are a group of enzymes that catalyze the phosphorylation of hydroxyl groups in specific residues of proteins, such as tyrosine, serine or threonine residues. Such phosphorylation may greatly alter the function of the protein; thus, protein kinases play an important role in the regulation of a wide variety of cellular processes, including, inter alia, metabolism, cell proliferation, cell differentiation, cell migration or cell survival. Among the various cellular functions involved in protein kinase activity, several processes are attractive targets for the treatment of cancer diseases and other diseases.

It is therefore an object of the present invention to provide compositions having anti-cancer activity which act, inter alia, on kinases. Among the kinases for which modulation of activity is sought, FAK, KDR and Tie2 are preferred.

These products conform to the following formula (I):

formula (I)

In the formula:

1) a and Ar are independently selected from: aryl, heteroaryl, heterocyclyl, substituted aryl, substituted heteroaryl, substituted heterocyclyl, cycloalkyl, substituted cycloalkyl;

2) l is selected from: NH, CO-NH, NH-CO, NH-SO₂，SO₂NH，NH-CH₂，CH₂-NH，CH₂-CO-NH，NH-CO-CH₂，NH-CH₂-CO，CO-CH₂-NH，NH-CO-NH，NH-CS-NH，NH-CO-O，O-CO-NH，CH₂-NH-CO-NH。NH-CO-NH-CH₂，NH-CO-CH₂-CO-NH；

3) Ra is selected from H, alkyl and cycloalkyl;

4) r1 is selected from: H. r, COR, SO₂R, wherein R is selected from H, OR "₄、NR”₅R”₆(C1-C6) alkyl, cycloalkyl, heterocyclyl, substituted heterocyclyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, wherein R "4 is selected from H, phenyl, alkyl, and wherein R" 5 and R "6 are independently selected from: H. ROR'₄(C1-C6) alkyl, cycloalkyl, heterocyclyl, substituted heterocyclyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, or R "5 and R" 6 are linked to each other to form a 5-to 8-mer (cha)non) saturated ring containing 0-3 heteroatoms selected from O, S and N;

5) r2 and R5 are independently selected from: H. halogen, R '2, CN, O (R' 2), OC (O) N (R '2) (R' 3), OS (O)₂)(R’2)，N(R’2)(R’3)，N＝C(R’2)(R’3)，N(R’2)C(O)(R’3)，N(R’2)C(O)O(R’3)，N(R’4)C(O)N(R’2)(R’3)，N(R’4)C(S)N(R’2)(R’3)，N(R’2)S(O₂)(R’3)，C(O)(R’2)，C(O)O(R’2)，C(O)N(R’2)(R’3)，C(＝N(R’3))(R’2)，C(＝N(OR’3))(R’2)，S(R’2)，S(O)(R’2)，S(O₂)(R’2)，S(O₂)O(R’2)，S(O₂)N(R’2) (R' 3); wherein each R ' 2, R ' 3, R ' 4 is independently selected from: H. alkyl, alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl, heterocyclyl, substituted alkyl, substituted alkenyl, substituted alkynyl, substituted aryl, substituted heteroaryl, substituted cycloalkyl, substituted heterocyclyl; wherein R '2 and R' 3 are each other than H and when both are at R2 or at R3, they are linked to each other to form a ring containing 0 to 3 heteroatoms selected from O, S and N.

Preferred products of the formula (I) correspond to the following definitions:

formula (I)

In the formula:

1) a and Ar are as defined above;

2) r1 is H;

3) l is selected from: NHCO, NH-CO-NH, NHSO₂、NHCO-CH₂-CONH；

4) Ra is selected from H and methyl;

5) r2 and R5 are as previously defined.

In the products of formula (I), Ar-L-A is advantageously:

wherein X1, X2, X3 and X4 are each independently selected from N and C-R '5, wherein R' 5 has the same definition as R2.

The substituent R' 5 is selected from H, F, Cl, methyl and NH₂、OMe、OCF₃And CONH₂Is preferred.

Preference is given toThe substituents R2 and R5 are independently selected from H, halogen, R ' 2, OR ' 2, NHR ' 2, NHCOR ' 2, NHCONHR ' 2, NHSO₂R' 2. R2 and R5 are preferably H.

The preferred substituent Ra is H.

Preferred substituents L-A are advantageously selected from NH-CO-NH-A and NH-SO₂-A。

A particularly effective combination L-A is obtained when L-A is NHCONH-A.

The products of the invention preferably have substituents A selected from the group consisting of phenyl, pyridyl, pyrimidinyl, thienyl, furyl, pyrrolyl, oxazolyl, thiazolyl, isoxazolyl, isothiazolyl, pyrazolyl, imidazolyl, indolyl, indazolyl, benzimidazolyl, benzoxazolyl and benzothiazolyl; optionally substituted.

More preferably, a is selected from phenyl, pyrazolyl and isoxazolyl; optionally substituted.

The substituent A is very advantageously substituted by a first substituent selected from alkyl, alkenyl, alkynyl, aryl, heteroaryl, O-alkyl, O-aryl, O-heteroaryl, S-alkyl, S-aryl, S-heteroaryl, each optionally substituted by a substituent selected from (C1-C3) alkyl, halogen, O- (C1-C3) alkyl.

Substituent A is preferably substituted with a second substituent selected from F, Cl, Br, I, OH, SH, SO₃M、COOM、CN、NO₂CON (R8) (R9), N (R8) CO (R9), (C1-C3) alkyl-OH, (C1-C3) alkyl-N (R8) (R9), (C1-C3) alkyl- (R10), (C1-C3) alkyl-COOH, N (R8) (R9); wherein R8 and R9 are independently selected from H, (C1-C3) alkyl, halo (C1-C3) alkyl, (C1-C3) alkyl OH, (C1-C3) alkyl NH₂(C1-C3) alkyl COOM, (C1-C3) alkyl SO₃M; wherein R8 and R9 are not H at the same time, are linked to form a 5-to 7-membered ring containing 0-3 heteroatoms; wherein M is H or an alkali metal cation selected from Li, Na and K; and wherein R10 is H or an optionally substituted non-aromatic heterocycle containing 2 to 7 carbon atoms and 1 to 3 heteroatoms selected from N, O and S.

Particularly preferred substituents A are selected from phenyl, pyrazolyl and isoxazolyl; the substituent A can be substituted by halogen, (C1-C4) alkyl, halo (C1-C3) alkyl, O- (C1-C4) alkyl, S- (C1-C4) alkyl, halo O- (C1-C4) alkyl and halo S- (C1-C4) alkyl. When A is disubstituted, the two substituents of A may form a 5-to 7-membered ring which contains from 0 to 3 heteroatoms.

The products of examples 1 to 41 are the subject of the present invention.

The product of the invention may be in the form of:

1) achiral, or

2) Racemic, or

3) Is enriched in stereoisomers, or

4) Enriched in enantiomers;

and may optionally be salified.

The product of the invention can be used for the manufacture of a medicament for the treatment of pathological conditions, in particular cancer.

The invention also relates to therapeutic compositions comprising the products of the invention in combination with pharmaceutically acceptable excipients according to the chosen mode of administration. The pharmaceutical composition may be in solid, liquid or liposome form.

Among the solid compositions, powders, gelatin capsules and tablets may be cited. In oral dosage forms, solid dosage forms that protect against the acidic medium of the stomach may also be included. The carriers used in the solid dosage form consist essentially of inorganic carriers such as phosphates and carbonates, or organic carriers such as lactose, cellulose, starch, or polymers. The liquid dosage form consists of a solution, suspension or dispersion. They contain water, organic solvents (ethanol, NMP, etc.) or mixtures of surfactants and solvents or complexing agents and solvents as dispersing carriers.

The liquid dosage form should preferably be injectable and therefore have an acceptable formulation for such a use.

Routes of administration that are acceptable for injection include intravenous, intraperitoneal, intramuscular, and subcutaneous routes, with the intravenous route generally being preferred.

The dosage of the compounds of the present invention to be administered will be adjusted by the practitioner depending on the route of administration to the patient and the condition of the patient.

The compounds of the present invention may be administered alone or in combination with other anticancer agents. Among the possible combinations, mention may be made of:

alkylating agents, in particular cyclophosphamide, melphalan, ifosfamide, chlorambucil, busulfan, thiotepa, prednimustine, carmustine, lomustine, semustine, streptozotocin (steptozocin), decarbazine, temozolomide, procarbazine and hexamethylmelamine

Platinum derivatives, e.g. in particular cisplatin, carboplatin or olmesartan

Antibiotics, e.g. in particular bleomycin, mitomycin, dactinomycin

Antimicrotubule agents, such as in particular vinblastine, vincristine, vindesine, vinorelbine, taxoids (taxol and docetaxel)

Anthracyclines, e.g. in particular doxorubicin, daunorubicin, idarubicin, epirubicin, mitoxantrone, iosoxantrone

Group I and II topoisomerase inhibitors, e.g. etoposide, teniposide, amsacrine, irinotecan, topotecan and tomodex

Fluoropyrimidines, e.g. 5-fluorouracil, UFT, floxuridine

Cytidine analogues, e.g. 5-azacytidine, cytarabine, gemcitabine, 6-mercaptomine, 6-thioguanine

Adenosine analogues, e.g. pentostatin, cytarabine or fludarabine phosphate

Methotrexate and folinic acid

Enzymes and compounds, e.g. L-asparaginase, hydroxyurea, trans-retinoic acid, sulfamine, dexrazoxane, amifostine, herceptine and female or male hormones

Antiangiogenic agents, for example combretastatin derivatives like CA4P, charlcons or colchicines like ZD6126 and prodrugs thereof.

It is also possible to use radiation therapy in combination with the compounds of the invention. These treatments may be administered simultaneously, sequentially or sequentially. Such treatment should be adjusted by the practitioner according to the patient to be treated.

The products of the invention are useful as inhibitors of kinase catalytic reactions. FAK, KDR and Tie2 are kinases for which the products of the invention should be particularly useful as inhibitors.

The reason for choosing these kinases is as follows:

FAK

FAK is a cytoplasmic tyrosine kinase that plays an important role in the transcription of signals emitted by integrins (the cell adhesion heterodimer receptor group). FAK and integrin are co-localized in the peripheral structures of the membrane known as focal adhesions. It has been demonstrated that in many cell types, activation of tyrosine residues by FAK and its phosphorylation, and in particular tyrosine 397 by autophosphorylation, is dependent on the binding of integrins to their extracellular ligands, and thus cell adhesion is induced [ Kornberg L, et al, j. biol. chem., (33): 23439-442.(1992)]. Autophosphorylation of FAK tyrosine 397 occupies, through its SH2 domain, the binding site for another tyrosine kinase, Src [ Schaller et al mol.cell.biol., 14; 1680-; xing et al, "mol.cell.biol.," 5; 413-421, 1994]. Src can then phosphorylate FAK on tyrosine 925, thus recruiting the adaptor protein Grb2 and inducing in certain cells the activation of ras and MAP kinase pathways involved in controlling cell proliferation [ Schlaepfer et al Nature; 372: 786-791.1994; one of Schlaepfer et al, "prog.biophy.mol.biol., 71: 435-478.1999; one of the book, Schlaepfer and Hunter, J.biol.chem., 272: 13189-13195, 1997]. Activation of FAK may also induce jun NH 2-terminal kinase (JNK), signaling a pathway that leads cells to progress toward G1 of the cell cycle [ Oktay et al, j.cell.biol., 145; 1461-1469, 1999]. phosphatidylinositol-3-OH kinase (PI 3-kinase) also binds to FAK at tyrosine 397, and this interaction may be necessary for activating PI 3-kinase [ Chen and Guan, proc.nat.acad.sci.usa, 91: 10148-; ling et al, "j.cell.biochem," 73; 533-544, 1999]. FAK/Src complexes phosphorylate various substrates, such as paxillin and p130CAS [ Vuori et al mol.cell.biol., 16: 2606-2613, 1996].

Many findings support the hypothesis that inhibitors of FAK may be useful in the treatment of cancer. Some studies suggest that FAK may play a major role in cell proliferation and/or survival in vitro. For example, in CHO cells, some authors have demonstrated that p125FAK overexpression results in accelerated conversion of G1 to S, and thus p125FAK is thought to promote cell proliferation [ Zhao j. -H et al j.cellbiol., 143: 1997-2008, 1998]. Other authors have demonstrated that tumor cells treated with antisense oligonucleotides to FAK lose their adhesion and enter apoptosis (Xu et al, CellGrowth Differ, 4: 413-418, 1996). FAK has also been shown to promote cell migration in vitro. Thus, fibroblasts that lack expression of FAK (FAK knockout mice) have a round morphology, and lack of response of cell migration to chemotactic signals and suppression of these defects by FAK re-expression [ dj.sieg et al, j.cell science, 112: 2677-91, 1999]. C-terminal domain overexpression (FRNK) of FAK prevents adhesion cell expansion and reduces cell migration in vitro [ Richardson a. and Parsons j.t., Nature, 380: 538-540, 1996]. Overexpression of FAK in CHO, COS cells or in human astrocytoma cells facilitates the migration of these cells. The complexity of FAK in promoting cell proliferation and migration in many in vitro cell types suggests a possible role for FAK in the neoplastic process. Recent studies have indeed demonstrated that induction of FAK expression in human astrocytoma cells results in increased proliferation of tumor cells in vitro [ Cary l.a. et al, j.cell sci., 109: 1787-94, 1996; wang D et al, "j.cell sci," 113: 4221-4230, 2000]. Furthermore, immunohistochemical studies of human biopsies have demonstrated that FAK has been overexpressed in prostate, breast, thyroid, colon, melanoma, brain and lung cancers, and that the expression levels of FAK are directly associated with tumors with the most aggressive phenotypes [ Weiner TM, et al, Lancet, 342 (8878): 1024 + 1025, 1993; owens et al, Cancer Research, 55: 2752 2755, 1995; maung K. et al, "Oncogene", 18: 6824-6828, 1999; wang D et al, "j.cell sci," 113: 4221-4230, 2000].

KDR

KDR (kinase insert domain receptor), also known as VEGF-R2 (vascular endothelial growth factor receptor 2), is expressed only in endothelial cells. This receptor binds to the vascular growth factor VEGF and thus acts as a mediator of the transduction signal by activating its intracellular kinase domain. Direct inhibition of VEGF-R2 kinase activity can reduce the phenomenon of angiogenesis in the presence of exogenous VEGF (vascular endothelial growth factor) (Strawn et al, cancer research, 1996, Vol.56, p.3540-3545). This process is demonstrated in particular by means of the VEGF-R2 mutant (Millauer et al, cancer research, 1996, Vol.56, p.1615-1620). Apart from being associated with the angiogenic activity of VEGF, the VEGF-R2 receptor does not appear to have any other function in adults. Therefore, selective inhibitors of VEGF-R2 kinase activity should demonstrate only minimal toxicity.

In addition to this central role in this dynamic angiogenic process, several new results suggest that VEGF expression contributes to tumor cell survival following chemo-and radiotherapy, thus emphasizing the possible synergistic effects of KDR inhibitors with other agents (Lee et al cancer research, 2000, Vol.60, p.5565-5570).

Tie2

Tie-2(TEK) is a member of the tyrosine kinase receptor family, a specific member of endothelial cells. Tie2 is the first receptor with tyrosine kinase activity, and is known to be both an agonist (angiopoietin 1 or Ang1) which stimulates autophosphorylation and cell signaling of the receptor [ S.Davis et al (1996) cell 87, 1161-1169] and an antagonist (angiopoietin 2 or Ang2) [ P.C.Maisonpierre et al (1997) science 277, 55-60 ]. Angiogenin 1 can act synergistically with VEGF during the final stages of neonatal lymphatic production (neoangiogenisis) [ Asahara T. C. Res. (1998), 233-. Knockout experiments and transgenic manipulation of Tie2 or Ang1 expression resulted in angiogenesis-deficient animals [ D.J. Dumont et al (1994), "Genes Dev.," 8, 1897-. The attachment of Ang1 to its receptor results in autophosphorylation of the Tie2 kinase domain, which is important for neovascularisation and for recruitment and vascular interaction with pericytes and smooth muscle cells; these phenomena contribute to the maturation and stabilization of newly formed blood vessels [ P.C. Maison Pierre et al (1997), science 277, 55-60 ]. Lin et al, (1997), < J. Invest, 100, 8: 2072 and Lin P. (1998), PNAS 95, 8829 and 8834, have shown inhibition of tumor growth and angiogenesis and lung metastasis when adenovirus is infected or injected with the intracellular domain of Tie-2(Tek) in a model of the mammary tumors xenograms and melanoma.

Inhibitors of Tie2 may be useful in situations where neovascularization is inappropriate (i.e., in diabetic retinopathy, chronic inflammation, psoriasis, Kaposi's sarcoma, chronic neovascularization due to macular degeneration, rheumatoid arthritis, infantile hemangiomas, and carcinomas).

Definition of

The term "halogen" refers to an element selected from the group consisting of F, Cl, Br and I.

The term "alkyl" refers to a straight or branched chain saturated hydrocarbon substituent having from 1 to 12 carbon atoms. Methyl, ethyl, propyl, 1-methylethyl, butyl, 1-methylpropyl, 2-methylpropyl, 1, 1-dimethylethyl, pentyl, 1-methylbutyl, 2-methylbutyl, 3-methylbutyl, 1, 1-dimethylpropyl, 1, 2-dimethylpropyl, 2, 2-dimethylpropyl, 1-ethylpropyl, hexyl, 1-methylpentyl, 2-methylpentyl, 1-ethylbutyl, 2-ethylbutyl, 3, 3-dimethylbutyl, heptyl, 1-ethylpentyl, octyl, nonyl, decyl, undecyl and dodecyl substituents are examples of alkyl substituents.

The term "alkenyl" refers to straight or branched hydrocarbon substituents having one or more unsaturations and from 2 to 12 carbon atoms. Vinyl, 1-methylvinyl, prop-1-enyl, prop-2-enyl, Z-1-methylprop-1-enyl, E-1-methylprop-1-enyl, Z-1, 2-dimethyl-prop-1-enyl, e-1, 2-dimethylprop-1-enyl, but-1, 3-dienyl, 1-methylidenyl-prop-2-enyl, Z-2-methylbut-1, 3-dienyl, E-2-methylbut-1, 3-dienyl, 2-methyl-1-methylidenyl-prop-2-enyl, undec-1-enyl and undec-10-enyl substituents are examples of alkenyl substituents.

The term "alkynyl" refers to straight or branched hydrocarbon substituents having at least two unsaturations carried by a pair of adjacent carbon atoms and from 2 to 12 carbon atoms. Ethynyl, prop-1-ynyl, prop-2-ynyl; and but-1-ynyl substituents are examples of alkynyl substituents.

The term "aryl" refers to a mono-or polycyclic aromatic substituent having from 6 to 14 carbon atoms. Examples of aryl substituents include phenyl, naphthalen-1-yl, naphthalen-2-yl, anthracen-9-yl, 1, 2, 3, 4-tetrahydronaphthalen-5-yl and 1, 2, 3, 4-tetrahydronaphthalen-6-yl substituents.

The term "heteroaryl" refers to a mono-or polycyclic heteroaromatic substituent having 1 to 13 carbon atoms and 1 to 4 heteroatoms. Pyrrol-1-yl, pyrrol-2-yl, pyrrol-3-yl, furyl, thienyl, imidazolyl, oxazolyl, thiazolyl, isoxazolyl, isothiazolyl, 1, 2, 4-triazolyl, oxadiazolyl, thiadiazolyl, tetrazolyl, pyridyl, pyrimidinyl, pyrazinyl, 1, 3, 5-triazinyl, indolyl, benzo [ b ] furyl, benzo [ b ] thienyl, indazolyl, benzimidazolyl, azaindolyl, quinolinyl, isoquinolinyl, carbazolyl and acridinyl substituents are examples of heteroaryl substituents.

The term "heteroatom" herein refers to at least a divalent atom that is not carbon. N, O, S and Se are examples of heteroatoms.

The term "cycloalkyl" refers to a saturated or partially unsaturated cyclic hydrocarbon substituent having from 3 to 12 carbon atoms. Cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclopentadienyl, cyclohexyl, cyclohexenyl, cycloheptyl, bicyclo [2.2.1] heptyl, cyclooctyl, bicyclo [2.2.2] octyl, adamantyl and perhydronaphthyl substituents are examples of cycloalkyl substituents.

The term "heterocyclyl" refers to a saturated or partially unsaturated cyclic hydrocarbon substituent having from 1 to 13 carbon atoms and from 1 to 4 heteroatoms. Preferably, the saturated or partially unsaturated cyclic hydrocarbon substituent should be monocyclic and should contain 4 or 5 carbon atoms and 1 to 3 heteroatoms.

The term "substituted" refers to one or more substituents other than H, such as halogen, alkyl, aryl, heteroaryl, cycloalkyl, heterocyclyl, alkenyl, alkynyl, OH, O-alkyl, O-alkenyl, O-aryl, O-heteroaryl, NH₂NH-alkyl, NH-aryl, NH-heteroaryl, N-alkyl', SH, S-alkyl, S-aryl, S (O)₂)H、S(O₂) Alkyl, S (O)₂) Aryl, SO₃H、SO₃Alkyl, SO₃Aryl, CHO, C (O) -alkyl, C (O) -aryl, C (O) OH, C (O) O-alkyl, C (O) O-aryl, OC (O) -alkyl, OC (O) -aryl, C (O) NH₂C (O) NH-alkyl, C (O) NH-aryl, NHCHO, NHC (O) -alkyl, NHC (O) -aryl, NH-cycloalkyl, NH-heterocyclyl.

The invention also relates to a method for producing the products of formula (I).

The products of the invention can be prepared by conventional organic chemistry methods.

Schemes 1, 2, 3 and 4 below illustrate the methods employed in the preparation of the examples involving the substituted pyrroles. In this regard, they are not intended to limit the scope of the invention with respect to the methods of preparing the claimed compounds.

Scheme 1

Scheme 2

Scheme 3

Scheme 4

It will be appreciated by those skilled in the art that in order to carry out the process of the invention described above, it may be necessary to introduce groups protecting the amino, carboxyl and alcohol functions in order to avoid side reactions. These groups are those that allow removal without affecting the rest of the molecule. As examples of groups protecting the amino function, mention may be made of tert-butyl carbamate, which can be regenerated with trifluoroacetic acid or iodotrimethylsilane, and acetyl, which can be regenerated in an acidic medium, for example hydrochloric acid. Examples of the group for protecting the carboxyl functional group include esters (e.g., methoxymethyl ester and benzyl ester). As the group protecting the alcohol function, there may be mentioned esters (for example, benzoyl esters) which can be regenerated in an acidic medium or by catalytic regeneration. Other protecting Groups that can be used are described in Protective Groups in Organic Synthesis, third edition, 1999, Wiley-Interscience, by GREENE et al.

The compound of formula (I) is isolated and may be purified by generally known methods, for example by crystallization, chromatography or extraction.

Enantiomers and diastereomers of the compounds of formula (I) are also part of the invention.

The compounds of formula (I) containing a basic residue can optionally be converted into addition salts with inorganic or organic acids by the action of such an acid in a solvent, for example an organic solvent such as an alcohol, ketone, ether or chlorinated solvent.

The compounds of formula (I) containing an acid residue may optionally be converted into metal salts or addition salts with nitrogen-containing bases according to methods known per se. These salts may be obtained by the action of a metal (e.g. alkali or alkaline earth metal) base, ammonia, amine or amine salt with a compound of formula (I) in a solvent. The salt formed is isolated by conventional methods.

These salts are also part of the present invention.

When the product of the invention has at least one free basic functional group, pharmaceutically acceptable salts can be prepared by reaction of the product with an inorganic or organic acid. Pharmaceutically acceptable salts include chloride, nitrate, sulfate, bisulfate, pyrosulfate, bisulfate, sulfite, bisulfite, phosphate, monohydrogen phosphate, dihydrogen phosphate, metaphosphate, pyrophosphate, acetate, propionate, acrylate, 4-hydroxybutyrate, caprylate, hexanoate, decanoate, oxalate, malonate, succinate, glutarate, adipate, pimelate, maleate, fumarate, citrate, tartrate, lactate, phenylacetate, mandelate, sebacate, suberate, benzoate, phthalate, methanesulfonate, p-toluenesulfonate, propanesulfonate, xylenesulfonate, salicylate, cinnamate, glutamate, aspartate, glucuronate, galacturonate.

Where the product of the invention has at least one free acidic functional group, pharmaceutically acceptable salts may be prepared by reaction of the product with an inorganic or organic base. Pharmaceutically acceptable bases include alkali metal or alkaline earth metal cation hydroxides, such as Li, Na, K, Mg, Ca, compounds containing basic amines, such as ammonia, arginine, histidine, piperidine, morpholine, piperazine, triethylamine.

The invention is also described by the following examples illustrating the invention.

LC/MS analysis was performed using a Micromass LCT model instrument in combination with an HP 1100 instrument. The abundance of the product was measured using an HP G1315A diode array detector in the wavelength range 200-600nm and a light scattering detector Sedex 65. Mass spectra were collected at 180-. This data was analyzed using the Micromass MassLynx software. The separation was performed using Hypersil BDS C18, 3 μm column (50X 4.6mm), eluting with a linear gradient of acetonitrile containing 0.05% (v/v) trifluoroacetic acid (TFA) 5-90% in water containing 0.05% (v/v) TFA at a flow rate of 1ml/min for 3.5 min. The total analysis time, including column reequilibration time, was 7 minutes.

Electrospray (ES) using a PlatformII device (Micromass)⁺) MS spectroscopy was performed. The major ions observed are described.

Melting points were measured by capillary method using a Mettler FP62 instrument at a temperature range of 30 deg.C-300 deg.C at 2 deg.C/min.

Purification by LC/MS:

the product can be purified by LC/MS using a Waters FractionsLynx system consisting of a Waters model 600 gradient pump, a Waters model515 regeneration pump, a Waters reagent manager dilution pump, a Waters model 2700 auto-injector, two Rheodyne modelsLabPro valve, Waters model 996 diode array detector, Waters model ZMD Mass spectrometer and Gilson model 204 fraction collector. The system was controlled with Waters FractionLynx software. Two Waters Symmetry columns (C) were used₁₈5 μm, 19X 50mm, ref 186000210) was alternated with one column being in regeneration with a water/acetonitrile 95/5(v/v) mixture containing 0.07% (v/v) trifluoroacetic acid and the other column being in separation. Elution of these columns was carried out using a linear gradient of 5-95% acetonitrile containing 0.07% (v/v) trifluoroacetic acid in water containing 0.07% (v/v) trifluoroacetic acid at a flow rate of 10 ml/min. At the exit of the separation column, one thousandth of the effluent was separated using an LC Packing Accurate, diluted with methanol at a flow rate of 0.5ml/min and sent to these detectors, 75% of which was sent to the diode array detector and the remaining 25% to the mass spectrometer. The remaining effluent (999/1000) was sent to a fraction collector, from which it was removed as long as the desired product quality was not detected using the FractionLynx software. The expected molecular formula was supplied to the FractionLynx software and the mass signal detected corresponded to the ion [ M + H ]]⁺And/or [ M + Na]⁺The software triggers the collection of its products. In some cases, depending on the results of the analysis of LC/MS, it was detected that the corresponding [ M +2H]⁺⁺For strong ions, a value corresponding to half the calculated molecular weight (MW/2) is also provided to the FractionLynx software. Under these conditions, the ion [ M +2H ] was detected]⁺⁺And/or [ M + Na + H]⁺⁺Quality signals, this collection is also triggered. These products were collected in tar coated glass tubes. After collection, the solvent was evaporated using a SavantAES 2000 or Genevac HT8 centrifugal evaporator and the tube after evaporation of the solvent was weighed to determine the mass of the product.

EI/CI analysis(ii) a Direct addition (DCI ═ deposition of sample on filament)

Finnigan SSQ7000 mass spectrometer; the mass range m/z is 29-900; electron energy 70 eV; the source temperature is 70 ℃; the reactant gas CI ammonia; EI-electron impact ionization; CI-chemical ionization.

Electrospray analysis:(electrospray ionization: ES)⁺(ii) a Negative electric spraying: ES-)

LC-MS-DAD-ELSD in combination:

method A

MS：Waters-Micromass Platform II；LC(ii) a Agilent HP 1100; hypersil GOLD Thermo C18 column; 3X 50mm, 3 μm; eluent: water (with 0.1% formic acid) + acetonitrile gradient, 7 min; the flow rate is 0.8 ml/min; UV; DAD (. lamda. about.200 and 400 nm).

Method B

MS：Waters-Micromass QTOF-2；LC: agilent HP 1100; hypersil GOLD Thermo C18 column; 3X 50mm, 3 μm; eluent: water (with 0.1% formic acid) + acetonitrile gradient, 7 min; the flow rate is 0.9 ml/min; UV; DAD (. lamda. about.200 and 400 nm).

Method C

MS：Waters-Micromass ZQ；LC: agilent HP 1100; XBRIDGE waters c18 column; 3X 50mm, 2.5 μm; eluent: water (with 0.1% formic acid) + acetonitrile gradient, 7 min; the flow rate is 1.1 ml/min; UV; DAD (λ 254 nm).

¹H NMR spectrum, 400MHz using BRUKER AVANCE DRX-400 spectrometer, or 300MHz using BRUKER AVANCE DPX-300 spectrometer, chemical shifts (. delta.,. ppm) -in dimethyl sulfoxide solvent-d 6(DMSO-d6) referenced to 2.50ppm at temperature 303K.

Example 1

4- {4- [3- (2-fluoro-5-trifluoromethylphenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide

Under the temperature of about 20 ℃ and the argon atmosphere, 0.012cm³2-fluoro-5-trifluoromethylphenyl isocyanate and 0.012cm³Triethylamine was added to 0.017g (84.48mmol) of 4- (4-aminophenyl) -1H-pyrrole-3-carboxamide at 27cm³Suspension in tetrahydrofuran. After stirring at a temperature of about 20 ℃ for 20 hours, the reaction mixture was concentrated to dryness under reduced pressure (2.7kPa), and the resulting residue was purified by flash chromatography [ eluent: ethyl acetate/dichloromethane (95/5, by volume)]. After concentration of the fractions under reduced pressure, a yellow residue was obtained which was at 5cm³Stirring in dichloromethane, filtration and drying under reduced pressure (2.7kPa) gave 22mg of 4- {4- [3- (2-fluoro-5-trifluoromethylphenyl) ureido]Phenyl } -1H-pyrrole-3-carboxamide as light brown solid¹HNMR(300MHz，(CD₃)₂SO, - δ, ppm): 6.57-7.02 (very broad m: 2H); 6.85 (wide t, J ═ 2.5 Hz: 1H); 7.29 (broad t, J ═ 2.5 Hz: 1H); 7.33-7.43 (m; 5H); 7.49(dd, J ═ 10.5 and 8.5 Hz: 1H); 8.60(dd, J ═ 7.5 and 2.5 Hz: 1H); 9.31 (broad s: 1H); 9.58 (broad s: 1H); 11.2 (broad s: 1H); EI: 406 (M/z)⁺)，m/z＝205(C₈H₃NOF₄ ⁺)，m/z＝179(C₇H₄NF₄ ⁺) Fundamental peak, ES +: 407 m/z (MH)⁺)。

4- (4-aminophenyl) -1H-pyrrole-3-carboxamide may be prepared in the following manner:

0.07g (0.304mmol) of ethyl 4- (4-aminophenyl) -1H-pyrrole-3-carboxylate at 10cm³The suspension in 22% aqueous ammonium hydroxide solution was heated in an autoclave at a temperature of about 80 ℃ for 84 hours. After stopping heating and then reducing to ambient temperature and pressure, the reaction mixture was concentrated to dryness under reduced pressure (2.7kPa) to give an orange solid which was further purified by flash chromatography [ eluent: dichloromethane/methanol/acetonitrile (98/1/1, by volume)]. After concentrating the fractions under reduced pressure, the residue obtained is at 10cm³Stirring in diethyl ether, filtration and drying under reduced pressure (2.7kPa) gave 0.02g of 4- (4-aminophenyl) -1H-pyrrole-3-carboxylic acidAmine, as brown solid: EI: m/z is 201 (M)⁺) Basic peak, M/z 185 (M-NH)₂ ⁺)，m/z＝157(M-CONH₂ ⁺)。

Ethyl 4- (4-aminophenyl) -1H-pyrrole-3-carboxylate may be prepared in the following manner:

0.2g (0.769mmol) of ethyl 4- (4-nitrophenyl) -1H-pyrrole-3-carboxylate are added at a temperature of about 20 ℃ to 0.02g (0.188mmol) of 10% palladium on carbon in 15cm³Suspension in methanol. After hydrogenation in an autoclave at a temperature of about 25 ℃ for 20 hours under 3 bar of hydrogen, the reaction mixture is filtered and the catalyst is rinsed three times with 5cm each time with methanol³The filtrate was then concentrated to dryness under reduced pressure (2.7kPa) to give a residue at 10cm³Stirring in diethyl ether, then filtering and drying under reduced pressure (2.7kPa) gave 0.079g of 4- (4-aminophenyl) -1H-pyrrole-3-carboxylic acid ethyl ester as a brown solid; EI: 230 (M/z)⁺) Basic peak, M/z 202 (M-C)₂H₄ ⁺)，m/z＝157(M-C₂H₅O⁺)。

Ethyl 4- (4-nitrophenyl) -1H-pyrrole-3-carboxylate may be prepared in the following manner:

to a solution of 0.512g (12.8mmol) of 60% sodium hydride in mineral oil at 20cm under an argon atmosphere at a temperature of about 20 deg.C³To the suspension in diethyl ether, a mixture of 2.212g (10mmol) of ethyl 4-nitrocinnamate and 1.991g (10.2mmol) of tosylmethyl isocyanate was added dropwise at 18cm³Dimethyl sulfoxide and 36cm³Solution in ether mixture. After stirring at reflux for 1 hour, the reaction mixture was dissolved in 70cm³Water, 20cm³Saturated aqueous sodium chloride solution and 100cm³Ethyl acetate mixture. The aqueous phase used was 50cm³Extracting with diethyl ether, and extracting with dichloromethane twice, each time 75cm³. All organic phases were combined, dried over anhydrous sodium sulfate, filtered and concentrated to dryness under reduced pressure (2.7kPa) to give a black oil which was dissolved in 75cm³Water and 50cm³Ethyl acetate mixture. Acetic acid B is used for the aqueous phaseExtracting the ester twice, 50cm each time³. All organic phases were combined, dried over anhydrous sodium sulfate, filtered and concentrated to dryness under reduced pressure (2.7kPa) to give 2.82g of a black solid which was further purified by flash chromatography [ eluent: cyclohexane/Ethyl acetate (3/2, by volume)]. After concentration of these fractions under reduced pressure, 1.48g of an orange solid was obtained, which was purified by flash chromatography [ eluent: methylene dichloride]. After concentrating the fractions under reduced pressure, 0.78g of 4- (4-nitrophenyl) -1H-pyrrole-3-carboxylic acid ethyl ester is obtained in the form of a yellow solid; EI: 260 (M/z)⁺) Basic peak, M/z 215 (M-C)₂H₅O⁺)，m/z＝169(215-NO₂)。

Example 2

1-acetyl-2-amino-4- {4- [3- (2-fluoro-5-trifluoromethylphenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide:

heating to about 20 deg.C under argon atmosphere to about 0.575cm³A solution of 4M hydrochloric acid in dioxane was added to 0.06g (0.115mmol) of 2-amino-3-carbamoyl-4- {4- [3- (2-fluoro-5-trifluoromethylphenyl) ureido]Phenyl } pyrrole-1-carboxylic acid tert-butyl ester at 1.2cm³Dioxane and 1.2cm³In solution in a methanol mixture. After stirring at a temperature of about 50 ℃ for 15 hours, the reaction mixture was concentrated to dryness under reduced pressure (2.7kPa) to give 2-amino-3-carbamoyl-4- {4- [3- (2-fluoro-5-trifluoromethylphenyl) ureido]Phenyl } -1H-pyrrole hydrochloride, dissolved in 2.5cm³In ethyl acetate. Adding 0.01cm of water at about 20 deg.C under argon atmosphere³Triethylamine and 0.013cm³Acetic anhydride. After stirring for 1 hour at a temperature of about 20 ℃, a catalytic amount of DMAP was added, and then stirring was maintained for 30 minutes. The reaction mixture used was 5cm³Ethyl acetate was diluted. The organic phase was washed twice with 5cm of water each time³. All aqueous phases were combined and used 5cm³And (5) extracting with ethyl acetate. All organic phases were combined and used 5cm³The residue was washed with saturated aqueous sodium chloride solution, dried over anhydrous magnesium sulfate, filtered and concentrated to dryness under reduced pressure (2.7kPa), to give 0.054g of residue, which was purified by flash chromatography [ eluent: methylene chloride/methanol (98/2, by volume)]. After concentrating the fractions under reduced pressure, 0.010g of 1-acetyl-2-amino-4- {4- [3- (2-fluoro-5-trifluoromethylphenyl) ureido]Phenyl } -1H-pyrrole-3-carboxamide, in the form of a yellow solid;¹H NMR(300MHz，(CD₃)₂SO, - δ, ppm): 2.14 (s: 3H); 5.20-6.10 (m: 1H wide); 6.40(d, J ═ 2.0 Hz: 1H); 6.70-7.60 (m: 1H wide); 7.30 (wide d, J ═ 8.5 Hz: 2H); 7.38 (mt: 1H); 7.45-7.54 (m: 3H); 8.62(dd, J ═ 7.5 and 2.5 Hz: 1H); 8.92 (broad s: 1H); 9.25 (broad s: 1H); 10.7 (broad s: 1H); 11.4 (broad s: 1H); ES +: m/z 464 (MH)⁺)。

Tert-butyl 2-amino-3-carbamoyl-4- {4- [3- (2-fluoro-5-trifluoromethylphenyl) ureido ] phenyl } pyrrole-1-carboxylate may be prepared as follows:

to a solution of 0.07g (0.221mmol) of tert-butyl 2-amino-4- (4-aminophenyl) -3-carbamoylpyrrole-1-carboxylate at 2cm under argon at a temperature of about 20 deg.C³Adding 0.125cm of tetrahydrofuran solution³Triethylamine and 0.049cm³2-fluoro-5-trifluoromethylphenyl isocyanate. After stirring at a temperature of about 20 ℃ for 4 hours, the reaction mixture was dissolved in 5cm³In dichloromethane. The organic phase was washed twice with 5cm of water each time³. All aqueous phases were combined and used 5cm³And (4) extracting with dichloromethane. All organic phases were combined and used 5cm³The extract was washed with a saturated aqueous sodium chloride solution, dried over anhydrous magnesium sulfate, filtered and concentrated to dryness under reduced pressure (2.7 kPa). 0.135g of an orange solid is obtained, which is purified again by flash chromatography [ eluent: methylene chloride/methanol (100/0-98/2 by volume)]. After concentration of these fractions under reduced pressure, 0.077g of 2-amino-3-carbamoyl-4- {4- [3- (2-fluoro-5-trifluoromethylphenyl) -ureido]Phenyl } pyrrole-1-carboxylic acid tert-butyl ester as a yellow solid; ES +: 522 (MH) is defined as m/z⁺)。

Tert-butyl 2-amino-4- (4-aminophenyl) -3-carbamoylpyrrole-1-carboxylate may be prepared in the following manner:

to a solution of 0.008g (0.0076mmol) of 10% palladium on carbon at 12cm at a temperature of about 25 deg.C³To the suspension in methanol was added 0.075g (0.216mmol) of tert-butyl 2-amino-3-carbamoyl-4- (4-nitrophenyl) pyrrole-1-carboxylate. After hydrogenation in an autoclave under 3 bar of hydrogen at a temperature of about 25 ℃ for 17 hours, the reaction mixture is filtered and the catalyst is rinsed three times with 5cm each time with methanol³The filtrate was then concentrated to dryness under reduced pressure (2.7kPa) to give 0.072g of tert-butyl 2-amino-4- (4-aminophenyl) -3-carbamoylpyrrole-1-carboxylate as a yellow solid;¹HNMR(400MHz，(CD₃)₂SO, -delta, ppm); 1.55 (s: 9H); 4.80-5.35 (very broad m: 1H); 5.22 (broad s: 2H); 6.32 (s: 1H); 6.40-6.85 (very broad m: 1H); 6.60 (wide d, J ═ 8.5 Hz: 2H); 6.99 (wide d, J ═ 8.5 Hz: 2H); 7.01 (broad s: 2H).

Tert-butyl 2-amino-3-carbamoyl-4- (4-nitrophenyl) pyrrole-1-carboxylate may be prepared in the following manner:

to 0.11g (0.447mmol) of 2-amino-4- (4-nitrophenyl) -1H-pyrrole-3-carboxamide at 6cm under an argon atmosphere at a temperature of about 20 deg.C³Adding 0.075cm into the suspension in dichloromethane³(0.536mmol) triethylamine was added followed by 0.117g (0.536mmol) di-tert-butyl dicarbonate. After stirring at about 50 ℃ for 2.5 hours, 0.09g (0.412mmol) of di-tert-butyl dicarbonate was added and stirring was maintained at about 50 ℃ for 3 hours. The reaction mixture was dissolved in 5cm³In dichloromethane. The organic phase was washed three times with 5cm of water each time³. All aqueous phases were combined and used 5cm³And (4) extracting with dichloromethane. All organic phases were combined, dried over anhydrous magnesium sulfate, filtered and concentrated to dryness under reduced pressure (2.7kPa) to give 0.28g of residue which was purified by flash chromatography [ eluent: methylene dichloride]. After concentration of these fractions under reduced pressure, 0.075g of 2-amino-3-ammonia is obtainedCarbamoyl-4- (4-nitrophenyl) pyrrole-1-carboxylic acid tert-butyl ester, in the form of an orange solid;¹HNMR(300MHz，(CD₃)₂SO, - δ, ppm): 1.57 (s; 9H); 5.77-6.58 (m: 2H wide); 6.72 (s: 1H); 6.93 (Wide s: 2H); 7.63 (wide d, J ═ 8.5 Hz: 2H); 8.23 (wide d, J ═ 8.5 Hz: 2H).

2-amino-4- (4-nitrophenyl) -1H-pyrrole-3-carboxamide may be prepared in the following manner:

0.26g (1.139mmol) 2-amino-4- (4-nitrophenyl) -1H-pyrrole-3-carbonitrile at 5cm³The suspension in concentrated sulfuric acid was heated at a temperature of about 80c for 1 hour. After cooling the reaction mixture to a temperature of about 20 ℃, it was poured onto crushed ice and then 5N aqueous sodium hydroxide was slowly added until a basic pH of about 10. The reaction mixture was extracted 7 times with a dichloromethane/MeOH mixture (98/2 by volume) of 10cm each time³. All organic phases were combined, dried over anhydrous magnesium sulfate, filtered and concentrated to dryness under reduced pressure (2.7kPa) to give 0.19g of 2-amino-4- (4-nitrophenyl) -1H-pyrrole-3-carboxamide as a red-brown solid; EI: 246 (M/z)⁺) Basic peak, M/z 229 (M-NH)₃ ⁺)。

2-amino-4- (4-nitrophenyl) -1H-pyrrole-3-carbonitrile may be prepared in the following manner:

to 0.5g (2.25 mmol) of N- [2- (4-nitrophenyl) -2-oxoethyl at a temperature of about 20 ℃ under an argon atmosphere]Acetamide at 15cm³To a solution in methanol was added 0.223g (3.375mmol) of malononitrile. The reaction medium is cooled to a temperature of about 0 ℃ and then 0.5cm of water is added³50% aqueous potassium hydroxide solution. After stirring at a temperature of about 0 ℃ for 15 minutes and then at a temperature of about 65 ℃ for 30 minutes, the reaction mixture was cooled to a temperature of about 20 ℃ and then poured onto crushed ice and extracted seven times with 10cm each of dichloromethane³. All organic phases were combined and used for 50cm³The extract was washed with saturated aqueous sodium chloride solution, dried over anhydrous magnesium sulfate, filtered and concentrated to dryness under reduced pressure (2.7kPa) to give 0.7g of brown solid, which was further purified by flash chromatography [ eluent: dichloro-methylAlkane/methanol (100/0-95/5 by volume)]. After concentrating the fractions under reduced pressure, 0.265g of 2-amino-4- (4-nitrophenyl) -1H-pyrrole-3-carbonitrile are obtained as a brown solid; EI: 228 (M/z)⁺) Basic peak, M/z 182 (M-NO)₂).

N- [2- (4-nitrophenyl) -2-oxoethyl ] acetamide may be prepared as follows:

to 0.5g (2.308mmol) of 2-amino- (4-nitrophenyl) acetophenone at 2cm under an argon atmosphere at a temperature of about 0 deg.C³Adding 0.435cm into suspension in water³(4.616mmol) acetic anhydride, then 0.379g (4.616mmol) sodium acetate in 1.5cm³A solution in water. After allowing the temperature to vary between 0 ℃ and 20 ℃ for 1 hour, 1.5cm was added³Concentrated hydrochloric acid until pH 2. The reaction mixture was extracted five times with 10cm each time with dichloromethane³. All organic phases were combined, dried over anhydrous magnesium sulfate, filtered and concentrated to dryness under reduced pressure (2.7kPa) to give 0.37g of N- [2- (4-nitrophenyl) -2-oxoethyl]Acetamide in a yellow solid form; ES +: 223 (MH) m/z⁺)。

Example 3

2-formylamino-4- {4- [3- (2-fluoro-5-trifluoromethylphenyl) ureido ] phenyl) -1H-pyrrole-3-carboxamide:

to 0.125g (0.578mmol) of 2-formylamino-4- (4-aminophenyl) -1H-pyrrole-3-carboxamide at 15cm at a temperature of about 23 deg.C³Adding 0.081cm into tetrahydrofuran solution³Triethylamine and 0.084cm³2-fluoro-5-trifluoromethylphenyl isocyanate. After stirring at a temperature of about 23 ℃ for 16 hours, the reaction mixture was concentrated to dryness under reduced pressure (2.7 kPa). The residue was re-diluted to 50cm³In ethyl acetate, then 50cm³Water and 50cm³The washing was carried out twice with a saturated aqueous solution of sodium chloride,50cm each time³. The organic solution was dried over anhydrous magnesium sulfate, treated with 3S carbon black, filtered and concentrated to dryness under reduced pressure (2.7kPa) to give 0.115g of an oil-like solid, which was further purified by flash chromatography [ eluent: dichloromethane/methanol/acetonitrile (95/2.5/2.5 by volume)]. After concentration of these fractions under reduced pressure, 0.016g of 2-formylamino-4- {4- [3- (2-fluoro-5-trifluoromethylphenyl) ureido]Phenyl } -1H-pyrrole-3-carboxamide, in the form of a cream solid, melts at 169 ℃.¹H NMR(400MHz，(CD₃)₂SO, - δ, ppm)5.60 (m, 1H wide); 6.41 (Wide s, 1H); 7.07 (m, 1H wide); 7.31(d, J ═ 8.5Hz, 2H); 7.38(m, 1H); 7.48(m partially masked, 1H); 7.51(d, J ═ 8.5Hz, 2H); 8.34(s, 1H); 8.61(dd, J ═ 2.5 and 7.5Hz, 1H); 9.07 (wide s, 1H); 9.40 (wide s, 1H); 10.85 (m, 1H wide); 11.45 (wide s, 1H).

2-formylamino-4- (4-aminophenyl) -1H-pyrrole-3-carboxamide may be prepared in the following manner:

to a solution of 0.030g (0.0285mmol) of 10% palladium on carbon at 15cm at a temperature of about 25 deg.C³To the suspension in methanol was added 0.23g (0.834mmol) of 2-formylamino-4- (4-nitrophenyl) -1H-pyrrole-3-carboxamide. After hydrogenation in an autoclave under 2 bar of hydrogen at a temperature of about 25 ℃ for 5 hours, the reaction mixture is filtered and the catalyst is rinsed three times with 5cm each time with methanol³The filtrate was then concentrated to dryness under reduced pressure (2.7kPa) to give 2-formylamino-4- (4-aminophenyl) -1H-pyrrole-3-carboxamide as a green solid.¹H NMR(400MHz，(CD₃)₂SO, - δ, ppm)5.13 (broad s, 2H); 5.43 (m, 1H wide); 6.27(d, J ═ 2.5Hz, 1H); 6.60(d, J ═ 9.0Hz, 2H); 6.80 (m, 1H wide); 6.99(d, J ═ 9.0Hz, 2H); 8.34(d, J ═ 1.5Hz, 1H); 10.9 (wide s, 1H); 11.25 (wide s, 1H).

2-formylamino-4- (4-nitrophenyl) -1H-pyrrole-3-carboxamide may be prepared in the following manner:

at a temperature near 25 ℃ to 0.3g (1.21mmol) of 2-Amino-4- (4-nitrophenyl) -1H-pyrrole-3-carboxamide at 5cm³Adding 2cm of the mixture into the solution in anhydrous ethanol³(52.9mmol) formic acid at 5cm³(52.9mmol) acetic anhydride. After stirring for 2 hours at this temperature, the reaction medium is poured into 100cm³In water. The suspension was then filtered. The solid was dehydrated and dried to give 0.257g of 2-formylamino-4- (4-nitrophenyl) -1H-pyrrole-3-carboxamide as a green powder.¹H NMR(400MHz，(CD₃)₂SO, - δ in ppm), 6.22-8.53 (very broad m, 2H); 6.78(d, J ═ 3.0Hz, 1H); 7.64(d, J ═ 9.0Hz, 2H); 8.19(d, J ═ 9.0Hz, 2H); 8.32(d, J ═ 1.5Hz, 1H); 10.55 (wide s, 1H); 11.6 (wide s, 1H).

2-amino-4- (4-nitrophenyl) -1H-pyrrole-3-carboxamide was prepared as described in example 2.

Example 4

2-isobutyrylamino-4- {4- [3- (2-fluoro-5-trifluoromethylphenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide:

to 0.125g (0.436mmol) of 2-isobutyrylamino-4- (4-aminophenyl) -1H-pyrrole-3-carboxamide at 15cm at a temperature of about 25 deg.C³Adding 0.076cm of tetrahydrofuran solution³(0.436mmol) 2-fluoro-5-trifluoromethylphenyl isocyanate. After stirring for 17 hours at a temperature of about 25 ℃, the reaction mixture was concentrated to dryness under reduced pressure (2.7 kPa). The residue was re-diluted at 40cm³In ethyl acetate, then using 40cm³And (4) washing with water. The organic solution was dried over anhydrous magnesium sulfate, filtered and concentrated to dryness under reduced pressure (2.7kPa) to give a residue at 8cm³Cyclohexane/ethyl acetate mixture (70/30 by volume). After filtration and drying, 0.096g of 2-isobutyrylamino-4- {4- [3- (2-fluoro-5-trifluoromethylphenyl) ureido]Phenyl } -1H-pyrrole-3-carboxamide, as a cream solid, melts at 196 ℃. IR (KBr), 3472, 3384, 1667, 1594, 1546, 1443, 1340, 1313, 1198, 1167, 1120, 1070, 937 and 614cm^-1。¹H NMR(400MHz，(CD₃)₂SO, - δ, ppm), 1.17(d, J ═ 7.0Hz, 6H); 2.61(m, 1H); 5.62 (m, 1H wide); 6.40(d, J ═ 2.5Hz, 1H); 7.00 (m, 1H wide); 7.30(d, J ═ 9.0Hz, 2H); 7.39(m, 1H); 7.49 (partially masked m, 1H); 7.51(d, J ═ 9.0Hz, 2H); 8.62(dd, J ═ 2.5 and 7.5Hz, 1H); 8.99 (wide s, 1H); 9.32 (wide s, 1H); 10.95 (wide s, 1H); 11.45 (wide s, 1H).

2-isobutyrylamino-4- (4-aminophenyl) -1H-pyrrole-3-carboxamide may be prepared in the following manner:

to 0.047g (0.0446mmol) of 10% palladium on carbon at 25cm at a temperature of about 25 deg.C³To the suspension in methanol was added 0.33g (1.04mmol) of 2-isobutyrylamino-4- (4-nitrophenyl) -1H-pyrrole-3-carboxamide. After hydrogenation in an autoclave at 2 bar of hydrogen at a temperature of about 25 ℃ for 3.5 hours, the reaction mixture is filtered and the catalyst is rinsed three times with 5cm each time with methanol³The filtrate was then concentrated to dryness under reduced pressure (2.7kPa) to give 0.22g of residue which was purified by flash chromatography [ eluent: dichloromethane/methanol/acetonitrile (96/2/2, by volume)]. After concentrating these fractions under reduced pressure, 0.135g of 2-isobutyrylamino-4- (4-aminophenyl) -1H-pyrrole-3-carboxamide is obtained in the form of a brown solid.¹H NMR(400MHz，(CD₃)₂SO, - δ, ppm)1.16(d, J ═ 7.0Hz, 6H); 2.60(m, 1H); 5.15 (wide s, 2H); 5.45 (m, 1H wide); 6.24(d, J ═ 2.0Hz, 1H); 6.60(d, J ═ 9.0Hz, 2H); 6.94 (m, 1H wide); 6.98(d, J ═ 9.0Hz, 2H); 11.05 (wide s, 1H); 11.3 (Wide s, 1H).

2-isobutyrylamino-4- (4-nitrophenyl) -1H-pyrrole-3-carboxamide may be prepared in the following manner:

to 0.23g (0.93mmol) of 2-amino-4- (4-nitrophenyl) -1H-pyrrole-3-carboxylic acid at a temperature close to 25 deg.CAmine at 15cm³Adding 0.260cm of tetrahydrofuran solution³(1.86mmol) Triethylamine and 0.098cm³(0.93mmol) isobutyryl chloride. After stirring at this temperature for 16 hours, the reaction medium is concentrated to dryness under reduced pressure (2.7kPa) to give a residue at 60cm³Stirring in water, extracting with ethyl acetate twice (50 cm each time)³. The organic phase was dried over anhydrous magnesium sulfate, filtered and concentrated to dryness under reduced pressure (2.7kPa) to give 0.35g of 2-isobutyrylamino-4- (4-nitrophenyl) -1H-pyrrole-3-carboxamide as a green powder.¹HNMR(400MHz，(CD₃)₂SO, - δ, ppm)15(d, J ═ 7.0Hz, 6H); 2.64(m, 1H); 6.50-8.50 (very broad m, 2H); 6.79(d, J ═ 2.5Hz, 1H); 7.64(d, J ═ 9.0Hz, 2H); 8.19(d, J ═ 9.0Hz, 2H); 10.45 (wide s, 1H); 11.65 (Wide s, 1H).

Example 5

2-butyrylamino-4- {4- [3- (2-fluoro-5-trifluoromethylphenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide:

to 0.207g (0.610mmol) of 2-butyrylamino-4- (4-aminophenyl) -1H-pyrrole-3-carboxamide at a temperature of about 25 ℃ in a volume of 20cm³Adding 0.096cm of tetrahydrofuran solution³(0.671mmol) 2-fluoro-5-trifluoromethylphenyl isocyanate. After stirring at a temperature of about 25 ℃ for 48 hours, the reaction mixture was concentrated to dryness under reduced pressure (2.7 kPa). The residue was re-diluted at 40cm³In ethyl acetate, then 30cm³Water and 30cm³The mixture was washed twice with saturated aqueous sodium chloride solution. The organic solution was dried over anhydrous magnesium sulfate, filtered and concentrated to dryness under reduced pressure (2.7kPa), and the resulting residue was purified by flash chromatography [ eluent: IIMethyl chloride/methanol/acetonitrile (97/1.5/1.5 by volume)]. After concentrating these fractions under reduced pressure, 0.093g of 2-butyrylamino-4- {4- [3- (2-fluoro-5-trifluoromethylphenyl) ureido]Phenyl } -1H-pyrrole-3-carboxamide, as a cream solid, melted at 219 ℃. IR (KBr), 3470, 3387, 1717, 1626, 1593, 1546, 1443, 1341, 1315, 1264, 1193, 1168, 1126, 1070, 820, 614cm^-1。¹H NMR(400MHz，(CD₃)₂SO, - δ, ppm)0.95(t, J ═ 7.5Hz, 3H); 1.64(m, 2H); 2.38(t, J ═ 7.5Hz, 2H); 5.60 (m, 1H wide); 6.39(d, J ═ 2.5Hz, 1H); 7.05 (m, 1H wide); 7.30(d, J ═ 9.0Hz, 2H); 7.38(m, 1H); 7.49 (partial mask m, 1H); 7.51(d, J ═ 9.0Hz, 2H); 8.61(dd, J ═ 2.5 and 7.5Hz, 1H); 9.05 (wide s, 1H); 9.38 (wide s, 1H); 10.8 (wide s, 1H); 11.4 (wide s, 1H).

2-butyrylamino-4- (4-aminophenyl) -1H-pyrrole-3-carboxamide may be prepared in the following manner:

to a volume of 0.068g (0.064mmol) of 10% palladium on carbon at 20cm at a temperature of about 25 deg.C³To the suspension in methanol was added 0.195g (0.61mmol) of 2-butyrylamino-4- (4-nitrophenyl) -1H-pyrrole-3-carboxamide. After hydrogenation in an autoclave under 2 bar of hydrogen at a temperature of about 25 ℃ for 5 hours, the reaction mixture is filtered and the catalyst is rinsed twice with 10cm each of methanol³The filtrate was then concentrated to dryness under reduced pressure (2.7kPa) to give 0.207g of 2-butyrylamino-4- (4-aminophenyl) -1H-pyrrole-3-carboxamide as a cream solid.¹H NMR(400MHz，(CD₃)₂SO, - δ, ppm)0.94(t, J ═ 7.5Hz, 3H); 1.63(m, 2H); 2.37(t, J ═ 7.5Hz, 2H); 5.11 (wide s, 2H); 6.05-8.45 (very broad m, 2H); 6.24(d, J ═ 2.5Hz, 1H); 6.60(d, J ═ 8.5Hz, 2H); 6.99(d, J ═ 8.5Hz, 2H); 10.8 (wide s, 1H); 11.3 (Wide s, 1H).

2-butyrylamino-4- (4-nitrophenyl) -1H-pyrrole-3-carboxamide may be prepared in the following manner:

at a temperature close to 25 DEG CAdd 0.20g (0.81mmol) of 2-amino-4- (4-nitro-phenyl) -1H-pyrrole-3-carboxamide at 25cm³Adding 0.227cm of tetrahydrofuran solution³(1.62mmol) Triethylamine and 0.085cm³(0.81mmol) butyryl chloride. 16 hours after stirring at this temperature, the reaction medium is concentrated to dryness under reduced pressure (2.7kPa) to give a residue at 60cm³Stirring in ethyl acetate, washing with water three times (20 cm each time)³. The organic phase was dried over anhydrous magnesium sulfate, filtered and concentrated to dryness under reduced pressure (2.7kPa) to give 0.221g of 2-butyrylamino-4- (4-nitrophenyl) -1H-pyrrole-3-carboxamide as a brown powder.¹HNMR(400MHz，(CD₃)₂SO, - δ, ppm)0.94(t, J ═ 7.5Hz, 3H); 1.63(m, 2H); 2.36(t, J ═ 7.5Hz, 2H); 6.20-8.50 (very broad m, 2H); 6.79(d, J ═ 3.0Hz, 1H); 7.64(d, J ═ 9.0Hz, 2H); 8.18(d, J ═ 9.0Hz, 2H); 10.3 (wide s, 1H); 11.6 (wide s, 1H).

Example 6

2- (3-cyclopentylpropionylamino) -4- {4- [3- (2-fluoro-5-trifluoromethylphenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide:

to 0.170g (0.50mmol) of 2- (3-cyclopentylpropionylamino) -4- (4-aminophenyl) -1H-pyrrole-3-carboxamide at 20cm at a temperature of about 25 deg.C³Adding 0.072cm into tetrahydrofuran solution³(0.50mmol) 2-fluoro-5-trifluoromethylphenyl isocyanate. After stirring at a temperature of about 25 ℃ for 48 hours, the reaction mixture was concentrated to dryness under reduced pressure (2.7 kPa). The residue was re-diluted to 50cm³In ethyl acetate, then 50cm³Water and 50cm³Washed with saturated aqueous sodium chloride solution. The organic solution was dried over anhydrous magnesium sulfateFiltered and concentrated to dryness under reduced pressure (2.7kPa), and the resulting residue was purified by flash chromatography [ eluent; dichloromethane/methanol/acetonitrile (95/2.5/2.5 by volume)]. After concentrating these fractions under reduced pressure, 0.048g of 2- (3-cyclopentylpropionylamino) -4- {4- [3- (2-fluoro-5-trifluoromethylphenyl) ureido]Phenyl } -1H-pyrrole-3-carboxamide, in the form of a yellow powder, which melts at 222 ℃. IR (KBr), 3470, 3389, 1717, 1633, 1594, 1546, 1443, 1341, 1312, 1194, 1167, 1119, 1070 and 614cm^-1。¹H NMR(400MHz，(CD₃)₂SO, - δ, ppm)1.11(m, 2H); 1.42-1.68(m, 6H); 1.71-1.85(m, 3H); 2.40(t, J ═ 8.0Hz, 2H); 5.56 (m, 1H wide); 6.39(d, J ═ 2.5Hz, 1H); 7.09 (m, 1H wide); 7.30(d, J ═ 9.0Hz, 2H); 7.38(m, 1H); 7.49 (partially masked m, 1H); 7.51(d, J ═ 9.0Hz, 2H); 8.62(dd, J ═ 2.5 and 7.5Hz, 1H); 9.00 (wide s, 1H); 9.32 (wide s, 1H); 10.8 (wide s, 1H); 11.4 (wide s, 1H).

2- (3-Cyclopentylpropionylamino) -4- (4-aminophenyl) -1H-pyrrole-3-carboxamide was prepared in the following manner:

to 0.055g (0.0051mmol) of 10% palladium on carbon at 25cm at a temperature of about 25 deg.C³To the suspension in methanol was added 0.210g (0.56mmol) of 2- (3-cyclopentylpropanoylamino) -4- (4-nitrophenyl) -1H-pyrrole-3-carboxamide. After hydrogenation in an autoclave under 2 bar of hydrogen at a temperature of about 25 ℃ for 5 hours, the reaction mixture is filtered and the catalyst is rinsed twice with 10cm each of methanol³The filtrate was then concentrated to dryness under reduced pressure (2.7kPa) to give 0.151g of 2- (3-cyclopentylpropanoylamino) -4- (4-aminophenyl) -1H-pyrrole-3-carboxamide as a brown powder.¹HNMR(400MHz，(CD₃)₂SO, - δ, ppm)1.09(m, 2H); 1.39-1.84(m, 9H); 2.39(t, J ═ 8.0Hz, 2H); 5.14 (wide s, 2H); 6.20-7.50 (very broad m, 2H); 6.23(d, J ═ 2.5Hz, 1H); 6.59(d, J ═ 8.5Hz, 2H); 6.99(d, J ═ 8.5Hz, 2H); 10.9 (wide s, 1H); 11.3 (Wide s, 1H).

2- (3-Cyclopentylpropionylamino) -4- (4-nitrophenyl) -1H-pyrrole-3-carboxamide may be prepared in the following manner:

to 0.15g (0.61mmol) of 2-amino-4- (4-nitrophenyl) -1H-pyrrole-3-carboxamide at 25cm at a temperature close to 25 deg.C³Adding 0.171cm of tetrahydrofuran solution³(1.22mmol) of triethylamine and 0.098mg (0.61mmol) of 3-cyclopentylpropionyl chloride. After stirring at this temperature for 16 hours, the reaction medium is concentrated to dryness under reduced pressure (2.7kPa) to give a residue at 40cm³Stirring in water, extracting with ethyl acetate three times (40 cm each time)³. The organic phase used was 80cm³The extract was washed with a saturated aqueous solution of sodium chloride, dried over anhydrous magnesium sulfate, filtered and concentrated to dryness under reduced pressure (2.7kPa) to give 0.218g of 2- (3-cyclopentylpropionylamino) -4- (4-nitrophenyl) -1H-pyrrole-3-carboxamide as a green powder.¹H NMR(400MHz，(CD₃)₂SO, - δ, ppm)1.11(m, 2H); 1.42-1.65(m, 6H); 1.69-1.86(m, 3H); 2.39(t, J ═ 8.0Hz, 2H); 6.17-8.50 (very broad m, 2H); 6.79(d, J ═ 2.5Hz, 1H); 7.63(d, J ═ 9.0Hz, 2H); 8.18(d, J ═ 9.0Hz, 2H); 10.35 (wide s, 1H); 11.6 (wide s, 1H).

Example 7

2- (cyclopropylcarbonylamino) -4- {4- [3- (2-fluoro-5-trifluoromethylphenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide

2- (Cyclopropylcarbonylamino) -4- {4- [3- (2-fluoro-5-trifluoromethylphenyl) ureido ] 2- (cyclopropylcarbonylamino) -4- {4- [3- (2-fluoro-5-trifluoromethylphenyl) ureido can be prepared as described in example 4 using 2-amino-4- (4-nitrophenyl) -1H-pyrrole-3-carboxamide and cyclopropylcarbonyl chloride]Phenyl } -1H-pyrrole-3-carboxamide. ES +: 490 (MH) for m/z⁺)。

Example 8

2-tert-pentanoylamino-4- {4- [3- (2-fluoro-5-trifluoromethylphenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide

Using 2-amino-4- (4-nitrophenyl) -1H-pyrrole-3-carboxamide and tert-pentanoyl chloride, 2-tert-pentanoylamino-4- {4- [3- (2-fluoro-5-trifluoromethylphenyl) ureido ] can be prepared as described in example 4]Phenyl } -1H-pyrrole-3-carboxamide. ES +: 506 (MH) m/z⁺)。

Example 9

2- (2-dimethylaminoacetylamino) -4- {4- [3- (2-fluoro-5-trifluoromethylphenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide

2- (2-Dimethylaminoacetylamino) -4- {4- [3- (2-fluoro-5-trifluoromethylphenyl) ureido ] can be prepared as described in example 4 using 2-amino-4- (4-nitrophenyl) -1H-pyrrole-3-carboxamide and dimethylglycine acid chloride]Phenyl } -1H-pyrrole-3-carboxamide ES +: 507 (MH) m/z⁺)。

Example 10

2-acetylamino-4- {6- [3- (2-fluoro-5-trifluoromethylphenyl) ureido ] pyridin-3-yl } -1H-pyrrole-3-carboxamide:

to 0.11g (0.424mmol) of 2-acetylamino-4- (6-aminopyridin-3-yl) -1H-pyrrole-3-carboxamide at a temperature of about 23 ℃ under an argon atmosphere at 20cm³Adding 0.068cm of tetrahydrofuran solution³(0.466mmol) 2-fluoro-5-trifluoromethylphenyl isocyanate. After stirring at a temperature of about 20 ℃ for 1 hour, 0.059cm was added to the medium³(0.424mmol) triethylamine. The reaction mixture was then stirred at this temperature for 18 hours and concentrated to dryness under reduced pressure (2.7 kPa). The residue was purified by flash chromatography [ eluent: dichloromethane/methanol (95/5, by volume) and an alcoholic ethyl acetate gradient]. After concentrating the fractions under reduced pressure, 0.013g of 2-acetylamino-4- {6- [3- (2-fluoro-5-trifluoromethylphenyl) ureido group was obtained]Pyridin-3-yl } -1H-pyrrole-3-carboxamide as a white solid. ES +: 466 (MH)⁺)。

2-acetylamino-4- (6-aminopyridin-3-yl) -1H-pyrrole-3-carboxamide may be prepared in the following manner:

to a solution of 0.015g (0.014mmol) of 10% palladium on carbon in 20cm at a temperature of about 25 deg.C³To the suspension in methanol was added 0.15g (0.519mmol) of 2-acetylamino-4- (6-nitropyridin-3-yl) -1H-pyrrole-3-carboxamide. After hydrogenation in 2 bar of hydrogen in an autoclave at a temperature of about 30 ℃ for 2 hours, the reaction mixture is filtered and the catalyst is rinsed twice with 2cm each time of diethyl ether³. After dehydration and drying, 0.11g of 2-acetylamino-4- (6-aminopyridin-3-yl) -1H-pyrrole-3-carboxamide is obtained in the form of a brown solid.¹H NMR(400MHz，(CD₃)₂SO, - δ, ppm)2.13(s, 3H); 6.01 (wide s, 2H); 5.50-8.85 (very broad m, 2H); 6.35(d, J ═ 2.5Hz, 1H); 6.49(d, J ═ 8.5Hz, 1H); 7.36(dd, J ═ 2.5 and 8.5Hz, 1H); 7.87(d, J ═ 2.5Hz, 1H); 10.65(s, 1H); 11.35 (wide s, 1H).

2-acetylamino-4- (6-nitropyridin-3-yl) -1H-pyrrole-3-carboxamide may be prepared in the following manner:

to 0.20g (0.809mmol) of 2-amino-4- (6-nitropyridin-3-yl) are added under an argon atmosphere at a temperature of approximately 20 deg.C) -1H-pyrrole-3-carboxamide at 40cm³Adding 0.226cm of tetrahydrofuran solution³(162mmol) Triethylamine and 0.058cm³(0.809mmol) acetyl chloride. After stirring at this temperature for 3 hours, the reaction medium is concentrated to dryness under reduced pressure (2.7kPa) and the residue obtained is diluted to 200cm³In ethyl acetate, then 50cm³Water and 50cm³The extract was washed with a saturated aqueous sodium chloride solution, dried over anhydrous magnesium sulfate, filtered and concentrated to dryness under reduced pressure (2.7kPa) to give 2g of acetylamino-4- (6-nitropyridin-3-yl) -1H-pyrrole-3-carboxamide as a green powder.¹H NMR(400MHz，(CD₃)₂SO, - δ, ppm)2.11(s, 3H); 6.90 (m, 2H wide); 6.95(d, J ═ 3.0Hz, 1H); 8.13(dd, J ═ 2.5 and 8.5Hz, 1H); 8.27(d, J ═ 8.5Hz, 1H); 8.65(d, J ═ 2.5Hz, 1H); 10.2(s, 1H); 11.7 (m, 1H wide).

2-amino-4- (6-nitro-3-pyridinyl) -1H-pyrrole-3-carboxamide may be prepared in the following manner:

to 0.17g (0.742mmol) of 2-amino-4- (6-nitro-3-pyridin-3-yl) -1H-pyrrole-3-carbonitrile was added 4.5cm at a temperature of about 5 deg.C³Concentrated sulfuric acid. The mixture was heated at a temperature of about 85 ℃ under an argon atmosphere for 1 hour. After cooling the reaction mixture to 5 ℃ it was poured onto crushed ice and 30cm³In water. Pouring the solution into 300cm³Tetrahydrofuran and 15cm³Pyridine solution. After stirring for 5 minutes, the organic phase took 30cm³Washed with saturated aqueous sodium chloride solution, then dried over anhydrous magnesium sulfate, filtered and concentrated to dryness under reduced pressure (2.7kPa) to give 0.20g of 2-amino-4- (6-nitropyridin-3-yl) -1H-pyrrole-3-carboxamide as a brown solid. ES +: 248 (MH/z)⁺)。

2-amino-4- (6-nitropyridin-3-yl) -1H-pyrrole-3-carbonitrile may be prepared in the following manner:

0.050g (0.224mmol) of N- [2- (6-nitropyridin-3-yl) -2-oxoethyl are charged at a temperature of about 20 ℃ under an argon atmosphere]Acetamide at 5cm³To a solution in methanol was added 0.022g (0.336mmol) of malononitrile. Reacting the mixtureThe medium was cooled to a temperature of about 0 ℃ and then 0.1cm was added³50% aqueous potassium hydroxide solution. After stirring at a temperature of about 20 ℃ for 15 minutes and then at a temperature of about 65 ℃ for 1.15 hours, the reaction mixture was concentrated to dryness under reduced pressure (2.7kPa) and then diluted at 50cm³In ethyl acetate. The organic phase used was 10cm³Water and 10cm³Washed with saturated aqueous sodium chloride solution, then dried over anhydrous magnesium sulfate, filtered and concentrated to dryness under reduced pressure (2.7kPa) to give 0.043g of 2-amino-4- (6-nitropyridin-3-yl) -1H-pyrrole-3-carbonitrile as a brown solid. ES +: 230 (MH) m/z⁺)。

N- [2- (6-Nitropyridin-3-yl) -2-oxoethyl ] acetamide may be prepared in the following manner:

to a volume of 1.05g (4.166mmol) of 2-amino-1- (6-nitropyridin-3-yl) ethanone at 25cm at a temperature of about 5 deg.C³Adding 1.575cm of water into the solution³(16.66mmol) acetic anhydride, then 1.367g (16.66mmol) sodium acetate in 3cm³A solution in water. After stirring for 3 hours at 5 ℃ the reaction mixture was extracted three times with 10cm each of ethyl acetate³. All organic phases were combined and used for 30cm³Washed with saturated aqueous sodium chloride solution, then dried over anhydrous magnesium sulfate, filtered and concentrated to dryness under reduced pressure (2.7kPa) to give 0.618g N- [2- (6-nitropyridin-3-yl) -2-oxoethyl]Acetamide in the form of a yellow solid.¹H NMR(400MHz，(CD₃)₂SO, - δ, ppm)1.90(s, 3H); 4.66(d, J ═ 4.5Hz, 2H); 8.37 (wide t, J ═ 4.5Hz, 1H); 8.43(d, J ═ 8.5Hz, 1H); 8.68(dd, J ═ 2.0 and 8.5Hz, 1H); 9.16(d, J ═ 2.0Hz, 1H).

2-amino-1- (6-nitropyridin-3-yl) ethanone can be prepared as follows:

to a solution of 0.975g (6.954mmol) hexamethylenetetramine in 10cm at a temperature of about 20 deg.C³To a solution of chlorobenzene in 1.549g (6.322mmol) of 2-bromo-1- (6-nitropyridin-3-yl) ethanone in 25cm³Solution in chlorobenzene. After stirring at this temperature for 1 hour, the suspension is heated at 50 ℃ for 18 hours, the reactionThe reaction medium is then cooled to 5 ℃ and then 200cm³And (4) diluting with diethyl ether. The precipitate thus obtained is filtered and washed three times with 50cm each of diethyl ether³. The obtained ammonium salt was found to be 20cm³Stirring in ethanol, and adding 8cm of ethanol at about 20 deg.C³37% hydrochloric acid. The solution was then stirred at this temperature for 16 hours. The precipitate formed is filtered and washed three times with 50cm of water each time³Dehydration and drying gave 1.05g 2-amino-1- (6-nitropyridin-3-yl) ethanone as a cream-colored powder. ES +: 182 (MH) m/z⁺)。

The 2-bromo-1- (6-nitropyridin-3-yl) ethanone can be prepared as follows:

at a temperature of about 20 ℃ 3.4g (20.46mmol) of 1- (6-nitropyridin-3-yl) ethanone are added at 60cm³To the solution in tetrahydrofuran, 7.28g (40.92mmol) of N-bromosuccinimide was added. After heating at reflux for 36 hours, the reaction mixture was concentrated to dryness under reduced pressure (2.7kPa), and then purified by flash chromatography [ eluent: pure methylene chloride]. After concentration of these fractions under reduced pressure, 1.7g of 2-bromo-1- (6-nitropyridin-3-yl) ethanone were obtained as a white powder. ES +: 246 (MH) m/z⁺)。

1- (6-nitropyridin-3-yl) ethanone can be prepared as follows:

to a solution of 1.044g (3.98mmol) of triphenylphosphine at 10cm under an argon atmosphere at a temperature of about 20 deg.C³To a solution in toluene were added 1.14g (1.99mmol) of bis (diphenylmethanone acetone) palladium and 10.1g (49.75mmol) of 5-bromo-2-nitropyridine. After stirring at this temperature for 15 minutes, 16.9cm were added³(49.75mmol) 1-ethoxyvinyltributyltin at 60cm³Solution in toluene. After 15 hours of heating under reflux, the reaction medium is cooled to 20 ℃ and then poured into 500cm³1N hydrochloric acid solution, stirred at this temperature for 16 hours. The medium is extracted with ethyl acetate three times, 150cm each time³. The organic phases were combined and then used for 300cm³Washing with water, drying over anhydrous magnesium sulfate, filtering and concentrating under reduced pressure (2.7kPa) to dryness, the residue obtained is subjected to flash chromatographyPurification by the method [ eluent: pure methylene chloride]. After concentration of these fractions under reduced pressure, 3.4g of 1- (6-nitropyridin-3-yl) ethanone are obtained.¹H NMR(400MHz，(CD₃)₂SO, - δ, ppm)2.71(s, 3H); 8.42(d, J ═ 8.5Hz, 1H); 8.66(dd, J ═ 2.5 and 8.5Hz, 1H); 9.15(d, J ═ 2.5Hz, 1H).

Example 11

2- (3-ethylureido) -4- {4- [3- (2-fluoro-5-trifluoromethylphenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide

To 0.056g (0.194mmol) of 4- (4-aminophenyl) -2- (3-ethylureido) -1H-pyrrole-3-carboxamide at a temperature of about 25 ℃ in a volume of 20cm³Adding 0.031cm into tetrahydrofuran solution³(0.0213mmol) 2-fluoro-5-trifluoromethylphenyl isocyanate. After stirring at a temperature of about 60 ℃ for 16 hours, the reaction mixture was concentrated to dryness under reduced pressure (2.7 kPa). The residue was re-diluted at 40cm³In ethyl acetate and then washed twice with 30cm each time of water³. The organic solution was dried over anhydrous magnesium sulfate, filtered and concentrated to dryness under reduced pressure (2.7kPa), and the resulting residue was purified by flash chromatography [ eluent: dichloromethane/methanol/acetonitrile (95/2.5/2.5 by volume)]. After concentrating the fractions under reduced pressure, 0.021g of 2- (3-ethylureido) -4- {4- [3- (2-fluoro-5-trifluoromethylphenyl) ureido]Phenyl } -1H-pyrrole-3-carboxamide, as a yellow solid. ES +: m/z 494 (MH)⁺)。

4- (4-aminophenyl) -2- (3-ethylureido) -1H-pyrrole-3-carboxamide may be prepared in the following manner:

to 0.067g (0.0636mmol) of 10% palladium on carbon at 15cm at a temperature of about 25 deg.C³To the suspension in methanol was added 0.115g (0.362mmol) of 4- (4-nitrophenyl) -2- (3-ethylureido) -1H-pyrrole-3-carboxamide. In a pressure kettleAfter hydrogenation in 2 bar of hydrogen at a temperature of about 25 ℃ for 2.5 hours, the reaction mixture is filtered and the catalyst is rinsed three times with 5cm each time in methanol³The filtrate was then concentrated to dryness under reduced pressure (2.7kPa) to give 0.056g of 4- (4-aminophenyl) -2- (3-ethylureido) -1H-pyrrole-3-carboxamide as an orange solid; ES +: m/z 289 (MH)⁺)。

4- (4-nitrophenyl) -2- (3-ethylureido) -1H-pyrrole-3-carboxamide may be prepared in the following manner:

to 0.2g (0.81mmol) of 2-amino-4- (4-nitrophenyl) -1H-pyrrole-3-carboxamide at 15cm at a temperature close to 25 deg.C³Adding 0.074cm of tetrahydrofuran solution³(0.891mmol) ethyl isocyanate and 0.005mg (0.040mmol) dimethylaminopyridine. After stirring for 16 hours at a temperature of approximately 60 ℃, the reaction medium is concentrated to dryness under reduced pressure (2.7kPa) to give a residue at 100cm³Stirring in water, extracting with ethyl acetate for three times (50 cm each time)³. The organic phase was dried over anhydrous magnesium sulfate, filtered and concentrated to dryness under reduced pressure (2.7kPa), and the resulting residue was purified by flash chromatography [ eluent: dichloromethane/methanol/acetonitrile (95/2.5/2.5 by volume)]. After concentration of these fractions under reduced pressure, 0.120g of 4- (4-nitrophenyl) -2- (3-ethylureido) -1H-pyrrole-3-carboxamide is obtained in the form of a yellow powder. ES +: 318 (MH) m/z⁺)。

Example 12

2-acetylamino-4- {4- [3- (3-fluoro-5-trifluoromethylphenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide

At room temperature, add 100mg (0.387mmol) of 2-acetylamino-4- (4-amino)Phenyl) -1H-pyrrole-3-carboxamide at 5cm³Adding 0.061cm into suspension in anhydrous tetrahydrofuran³(0.426mmol) 3-fluoro-5-trifluoromethylphenyl isocyanate. The reaction medium is stirred at room temperature for 24 hours, cooled in a water-ice bath and then filtered through a sintered glass filter. The collected solid was washed with a small amount of dichloromethane and cyclohexane and then dried in vacuo. This gave 75mg of 2-acetylamino-4- {4- [3- (3-fluoro-5-trifluoromethylphenyl) ureido]Phenyl } -1H-pyrrole-3-carboxamide, as a light brown solid.

LCMS (method a): m/z 464: [ M + H ]]⁺；m/z＝447：[M+H]⁺-NH₃(basic peak) m/z 462: [ M-H ]]^-. Retention time (m) was 3.97.

2-acetylamino-4- (4-aminophenyl) -1H-pyrrole-3-carboxamide may be prepared in the following manner:

add 1.36g (4.72mmol) 2-acetylamino-4- (4-nitrophenyl) -1H-pyrrole-3-carboxamide at 200cm³To the suspension in methanol, 0.15g of palladium on carbon (10%) was added. The hydrogenation is carried out at 2 bar and 30 ℃ for 6 hours, after which the reaction medium is filtered through kieselguhr and washed with methanol. The filtrate was evaporated under reduced pressure to give 1.14g of 2-acetylamino-4- (4-aminophenyl) -1H-pyrrole-3-carboxamide as a brown solid.¹H NMR(300MHz，(CD₃)₂SO d6- δ, ppm): 2.13(s, 3H); 5.13(s, 2H); 5.45 (m, 1H wide); 6.24(d, J ═ 2.5Hz, 1H); 6.59(d, J ═ 8.5Hz, 2H); 6.90-7.10 (m, 1H wide); 6.99(d, J ═ 8.5Hz, 2H); 10.8(s, 1H); 11.25 (wide s, 1H).

EI：m/z＝258：[M⁺](fundamental peak); m/z 241: [ M + H ]]⁺-NH₃；m/z＝199：241-COCH₃。

2-acetylamino-4- (4-nitrophenyl) -1H-pyrrole-3-carboxamide may be prepared in the following manner:

2.16g (8.77mmol) 2-amino-4- (4-nitrophenyl) -1H-pyrrole-3-Formamide at 200cm³0.851cm of a suspension in anhydrous tetrahydrofuran was added³(11.960mmol) acetyl chloride. The mixture was cooled with a water-ice bath and then 3.180cm was added slowly at 0 deg.C³(22.82mmol) triethylamine. The reaction was stirred at 0 ℃ for 15 minutes and then at room temperature for 3 hours. The medium is dissolved in ethyl acetate, and the organic phase is washed with water, then dried over magnesium sulfate, filtered and the solvent is distilled off under reduced pressure. The crude product was purified by flash chromatography [ eluent: methylene chloride/methanol (99/1, by volume)]. After concentration of the fraction containing the desired product under reduced pressure, 1.23g of 2-acetylamino-4- (4-nitrophenyl) -1H-pyrrole-3-carboxamide are obtained in the form of a brown solid.

¹H NMR(400MHz，(CD₃)₂SO d6- δ, ppm): 2.11(s, 3H); 6.50-7.20 (very broad m, 2H); 6.80(s, 1H); 7.63(d, J ═ 9.0Hz, 2H); 8.18(d, J ═ 9.0Hz, 2H); 10.3 (wide s, 1H); 11.6 (wide s, 1H).

LCMS (method a); 287 m/z: [ M-H ]]^-。

Retention time (m) 2.90.

2-amino-4- (4-nitrophenyl) -1H-pyrrole-3-carboxamide may be prepared as described in example 2, using 2-amino-4- (4-nitrophenyl) -1H-pyrrole-3-carbonitrile.

Example 13

2-acetylamino-4- {4- [3- (3-ethylphenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide

To the suspension 100mg (0.387mmol) of 2-acetylamino-4- (4-aminophenyl) -1H-pyrrole-3-carboxamide at 5cm at room temperature³Adding 0.061cm into solution of anhydrous tetrahydrofuran³(0.426mmol) 1-ethyl-3-isocyanatobenzene. The reaction medium is stirred at room temperature for 24 hours and then evaporated to dryness under reduced pressure. The crude product was purified by flash chromatography [ eluent; methylene chloride/methanol (97/3, by volume)]. The fraction containing the desired product was concentrated under reduced pressure to give 107mg of 2-acetylamino-4- {4- [3- (3-ethylphenyl) ureido]Phenyl } -1H-pyrrole-3-carboxamide. In the form of a brown solid.

LCMS (method a): 406/z: [ M + H ]]⁺

m/z＝389：[M+H]⁺-NH₃

Retention time (m) was 3.67.

Example 14

2-acetylamino-4- {4- [3- (4-fluoro-3-trifluoromethylphenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide

2-acetylamino-4- {4- [3- (4-fluoro-3-trifluoromethylphenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide may be prepared as described in example 13 using 4-fluoro-3- (trifluoromethyl) phenyl isocyanate and 2-acetylamino-4- (4-amino-phenyl) -1H-pyrrole-3-carboxamide. The method is characterized in that:

LCMS (method a): m/z 464: [ M + H ]]⁺；m/z447：[M+H]⁺-NH₃(fundamental peak);

m/z＝462：[M-H]^-，

retention time (m) was 3.83.

Example 15

2-acetylamino-4- {4- [3- (3-fluorophenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide

2-acetylamino-4- {4- [3- (3-fluorophenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide may be prepared as described in example 12 using 3-fluorophenyl isocyanate and 2-acetylamino-4- (4-aminophenyl) -1H-pyrrole-3-carboxamide. The method is characterized in that:

LCMS (method a): m/z 396: [ M + H ]]⁺；m/z＝379：[M+H]⁺-NH₃。

Retention time (m) 3.42.

Example 16

2-acetylamino-4- {4- [3- (4-fluorophenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide

2-acetylamino-4- {4- [3- (4-fluorophenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide may be prepared as described in example 12 using 4-fluorophenyl isocyanate and 2-acetylamino-4- (4-aminophenyl) -1H-pyrrole-3-carboxamide.

LCMS (method a): m/z 396: [ M + H ]]⁺；m/z＝379：[M+H]⁺-NH₃。

Retention time (m) 3.42.

Example 17

2-acetylamino-4- {4- [3- (2-fluorophenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide

2-Acetylamino-4- {4- [3- (2-fluorophenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide may be prepared as described in example 12 using 2-fluorophenyl isocyanate and 2-acetylamino-4- (4-aminophenyl) -1H-pyrrole-3-carboxamide.

LCMS (method B): m/z 396: [ M + H ]]⁺(fundamental peak); m/z 379: [ M + H ]]⁺-NH₃。

Retention time (m) was 3.70.

Example 18

2-acetylamino-4- {4- [3- (4-difluoromethoxyphenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide

2-acetylamino-4- {4- [3- (4-difluoromethoxy phenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide may be prepared as described in example 12 using 4- (difluoromethoxy) phenyl isocyanate and 2-acetylamino-4- (4-aminophenyl) -1H-pyrrole-3-carboxamide.

LCMS (method a): m/z 444: [ M + H ]]⁺；m/z 427：[M+H]⁺-NH₃

m/z＝442：[M-H]^-。

Retention time (m) 3.51.

Example 19

2-acetylamino-4- {4- [3- (3, 4-dimethylphenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide

2-acetylamino-4- {4- [3- (3, 4-dimethylphenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide may be prepared as described in example 12 using 3, 4-dimethylphenyl isocyanate and 2-acetylamino-4- (4-aminophenyl) -1H-pyrrole-3-carboxamide.

LCMS (method a): 406/z: [ M + H ]]⁺，m/z＝389：[M+H]⁺-NH₃。

m/z＝404：[M-H]^-

Retention time (m) 3.60.

Example 20

2-acetylamino-4- {4- [3- (3, 4-dimethoxyphenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide

2-acetylamino-4- {4- [3- (3, 4-dimethoxyphenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide may be prepared as described in example 12 using 3, 4-dimethoxyphenyl isocyanate and 2-acetylamino-4- (4-aminophenyl) -1H-pyrrole-3-carboxamide.

LCMS (method a): 438: [ M + H ]]⁺；m/z＝421：[M+H]⁺-NH₃

m/z＝436：[M-H]^-。

Retention time (m) 2.94.

Example 21

2-acetylamino-4- {4- [3- (4-trifluoromethoxyphenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide

2-acetylamino-4- {4- [3- (4-trifluoromethoxyphenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide may be prepared as described in example 12 using 4- (trifluoromethoxy) phenyl isocyanate and 2-acetylamino-4- (4-aminophenyl) -1H-pyrrole-3-carboxamide.

LCMS (method a): 462 of m/z: [ M + H ]]⁺；m/z＝445：[M+H]⁺-NH₃

m/z＝460：[M-H]^-。

Retention time (m) 3.85.

Example 22

2-acetylamino-4- {4- [3- (2, 5-dimethoxyphenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide

2-acetylamino-4- {4- [3- (2, 5-dimethoxyphenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide may be prepared as described in example 13 using 2, 5-dimethoxyphenyl isocyanate and 2-acetylamino-4- (4-aminophenyl) -1H-pyrrole-3-carboxamide.

LCMS (method C): 438: [ M + H ]]⁺。

m/z＝436：[M-H]^-。

Retention time (m) 3.16.

Example 23

2-acetylamino-4- [4- (3-phenylureido) phenyl ] -1H-pyrrole-3-carboxamide

2-Acetylamino-4- [4- (3-phenylureido) phenyl ] -1H-pyrrole-3-carboxamide may be prepared as described in example 13 using phenyl isocyanate and 2-acetylamino-4- (4-aminophenyl) -1H-pyrrole-3-carboxamide.

LCMS (method C): 378 for m/z: [ M + H ]]⁺。

m/z＝376：[M-H]^-。

Retention time (m) was 2.97.

Example 24

2-acetylamino-4- {4- [3- (2-methoxyphenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide

2-acetylamino-4- {4- [3- (2-methoxyphenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide may be prepared as described in example 13 using 2-methoxyphenyl isocyanate and 2-acetylamino-4- (4-aminophenyl) -1H-pyrrole-3-carboxamide.

LCMS (method C): m/z 408: [ M + H ]]⁺

m/z＝406：[M-H]^-

Retention time (m) 3.16

Example 25

2-acetylamino-4- {4- [3- (2-trifluoromethylphenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide

2-acetylamino-4- {4- [3- (2-trifluoromethylphenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide may be prepared as described in example 13 using 2- (trifluoromethyl) phenyl isocyanate and 2-acetylamino-4- (4-aminophenyl) -1H-pyrrole-3-carboxamide.

LCMS (method C): m/z 446: [ M + H ]]⁺

m/z＝444：[M-H]^-

Retention time (m) 3.32

Example 26

2-acetylamino-4- [4- (3-o-tolylureido) phenyl ] -1H-pyrrole-3-carboxamide

2-acetylamino-4- [4- (3-o-tolylureido) phenyl ] -1H-pyrrole-3-carboxamide may be prepared as described in example 13 using 2-methylphenyl isocyanate and 2-acetylamino-4- (4-aminophenyl) -1H-pyrrole-3-carboxamide.

LCMS (method C): 392 m/z: [ M + H ]]⁺

m/z＝390：[M-H]^-

Retention time (m) was 3.07.

Example 27

2-acetylamino-4- {4- [3- (3-methoxyphenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide

2-acetylamino-4- {4- [3- (3-methoxyphenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide may be prepared as described in example 13 using 3-methoxyphenyl isocyanate and 2-acetylamino-4- (4-aminophenyl) -1H-pyrrole-3-carboxamide.

LCMS (method C): m/z 408: [ M + H ]]⁺

m/z＝406：[M-H]^-

Retention time (m) 3.01

Example 28

2-acetylamino-4- {4- [3- (3-trifluoromethylphenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide

2-acetylamino-4- {4- [3- (3-trifluoromethylphenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide may be prepared as described in example 13 using 3- (trifluoromethyl) phenyl isocyanate and 2-acetylamino-4- (4-aminophenyl) -1H-pyrrole-3-carboxamide.

LCMS (method C): m/z 446: [ M + H ]]⁺

m/z＝444：[M-H]^-

Retention time (m) 3.54

Example 29

2-acetylamino-4- [4- (3-m-tolylureido) phenyl ] -1H-pyrrole-3-carboxamide may be prepared as described in example 13 using 3-methylphenyl isocyanate and 2-acetylamino-4- (4-aminophenyl) -1H-pyrrole-3-carboxamide.

LCMS (method C): 392 m/z: [ M + H ]]⁺

m/z＝390：[M-H]^-

Retention time (m) 3.20

Example 30

2-acetylamino-4- {4- [3- (4-trifluoromethylphenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide

2-acetylamino-4- {4- [3- (4-trifluoromethylphenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide may be prepared as described in example 13 using 4- (trifluoromethyl) phenyl isocyanate and 2-acetylamino-4- (4-aminophenyl) -1H-pyrrole-3-carboxamide.

LCMS (method C): m/z 446: [ M + H ]]⁺

m/z＝444：[M-H]^-

Retention time (m) 3.58

Example 31

2-acetylamino-4- [4- (3-p-tolylureido) phenyl ] -1H-pyrrole-3-carboxamide may be prepared as described in example 13 using 4-methylphenyl isocyanate and 2-acetylamino-4- (4-aminophenyl) -1H-pyrrole-3-carboxamide.

LCMS (method C): 392 m/z: [ M + H ]]⁺

m/z＝390：[M-H]^-

Retention time (m) 3.19

Example 32

2-acetylamino-4- {4- [3- (4-chloro-3-trifluoromethylphenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide

2-acetylamino-4- {4- [3- (4-chloro-3-trifluoromethylphenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide may be prepared as described in example 13 using 4-chloro-3- (trifluoromethyl) phenyl isocyanate and 2-acetylamino-4- (4-aminophenyl) -1H-pyrrole-3-carboxamide.

LCMS (method C): m/z 480: [ M + H ]]⁺

m/z 478＝[M-H]^-

Retention time (m) 3.80

Example 33

2-acetylamino-4- {4- [3- (2-chloro-5-trifluoromethylphenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide

2-acetylamino-4- {4- [3- (2-chloro-5-trifluoromethylphenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide may be prepared as described in example 13 using 2-chloro-5- (trifluoromethyl) phenyl isocyanate and 2-acetylamino-4- (4-aminophenyl) -1H-pyrrole-3-carboxamide.

LCMS (method C): m/z 480: [ M + H ]]⁺

m/z＝478：[M-H]^-

Retention time (m) 3.82

Example 34

2-acetylamino-4- {4- [3- (2-fluoro-3-trifluoromethylphenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide

2-acetylamino-4- {4- [3- (2-fluoro-3-trifluoromethylphenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide may be prepared as described in example 13 using 2-fluoro-3- (trifluoromethyl) phenyl isocyanate and 2-acetylamino-4- (4-aminophenyl) -1H-pyrrole-3-carboxamide.

LCMS (method C): m/z 464: [ M + H ]]⁺

m/z＝462：[M-H]^-

Retention time (m) 3.64

Example 35

2-acetylamino-4- {4- [3- (3-chloro-4-difluoromethoxyphenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide

2-acetylamino-4- {4- [3- (3-chloro-4-difluoromethoxyphenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide may be prepared as described in example 13 using 3-chloro-4- (di-fluoromethoxy) phenyl isocyanate and 2-acetylamino-4- (4-aminophenyl) -1H-pyrrole-3-carboxamide.

LCMS (method C): m/z 478: [ M + H ]]⁺

m/z＝476：[M-H]^-

Retention time (m) 3.52

Example 36

2-acetylamino-4- {4- [3- (3, 5-dimethoxyphenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide

2-acetylamino-4- {4- [3- (3, 5-dimethoxyphenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide may be prepared as described in example 13 using 3, 5-dimethoxyphenyl isocyanate and 2-acetylamino-4- (4-aminophenyl) -1H-pyrrole-3-carboxamide.

LCMS (method C): 438: [ M + H ]]⁺

m/z＝436：[M-H]^-

Retention time (m) 3.07

Example 37

2-acetylamino-4- {4- [3- (3, 5-dimethylphenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide

2-acetylamino-4- {4- [3- (3, 5-dimethylphenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide may be prepared as described in example 13 using 3, 5-dimethylphenyl isocyanate and 2-acetylamino-4- (4-aminophenyl) -1H-pyrrole-3-carboxamide.

LCMS (method C): 406/z: [ M + H ]]⁺

m/z＝404：[M-H]^-

Retention time (m) 3.43

Example 38

2-acetylamino-4- {4- [3- (2, 5-dimethylphenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide

2-acetylamino-4- {4- [3- (2, 5-dimethylphenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide may be prepared as described in example 13 using 2, 5-dimethylphenyl isocyanate and 2-acetylamino-4- (4-aminophenyl) -1H-pyrrole-3-carboxamide.

LCMS (method C): 406/z: [ M + H ]]⁺

m/z＝404：[M-H]^-

Retention time (m) 3.29

Example 39

2-acetylamino-4- {4- [3- (2-methoxy-5-methylphenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide

2-acetylamino-4- {4- [3- (2-methoxy-5-methylphenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide may be prepared as described in example 13 using 2-methoxy-5-methylphenyl isocyanate and 2-acetylamino-4- (4-aminophenyl) -1H-pyrrole-3-carboxamide.

LCMS (method C): m/z 422: [ M + H ]]⁺

m/z＝420：[M-H]^-

Retention time (m) 3.38

Example 40

2-acetylamino-4- {4- [3- (4-methyl-3-trifluoromethylphenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide

2-acetylamino-4- {4- [3- (4-methyl-3-trifluoromethylphenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide may be prepared as described in example 13 using 3- (trifluoromethyl) -4-methylphenyl isocyanate and 2-acetylamino-4- (4-aminophenyl) -1H-pyrrole-3-carboxamide.

The melting point was 266 ℃.

ES⁺：m/z＝460：[M+H]⁺；m/z＝482：[M+Na]⁺

EXAMPLE 41

2-acetylamino-4- [4- (2, 3-dichlorobenzenesulfonylamino) phenyl ] -1H-pyrrole-3-carboxamide

70mg (0.271mmol) of 2-acetylamino-4- (4-aminophenyl) -1H-pyrrole-3-carboxamide and 70mg (0.285mmol) of 2, 3-dichlorobenzenesulfonyl chloride at 1.5cm³The suspension in pyridine is stirred at room temperature for 24 hours, and the reaction medium is then evaporated to dryness under reduced pressure. The crude product was purified by flash chromatography [ eluent; methylene chloride/methanol (97/3, by volume)]. The fraction containing the desired product was concentrated under reduced pressure to give 22mg of 2-acetylamino-4- [4- (2, 3-dichlorophenylsulfonylamino) phenyl group]-1H-pyrrole-3-carboxamide, in the form of a light brown solid, melting at 265 ℃.

ES⁺：m/z＝469：[M+H]⁺

Determination of Compound Activity-Experimental protocol

1.FAK

The inhibitory activity of the compounds on FAK was determined by measuring the inhibition of autophosphorylation of the enzyme using a time resolved fluorescence assay (HTRF).

The N-terminal of the DNA fragment was a histidine-tagged cDNA of human FAK, which was cloned into the baculovirus expression vector pFastBac HTc. The protein was expressed and purified to about 70% homogeneity.

This enzyme (6.6. mu.g/ml) was mixed with various concentrations of the compound to be tested in a solution containing 10mM MgCl2, 100. mu.M Na₃VO₄The kinase activity was measured by culturing 15. mu.M ATP in 50mM Hepes buffer at pH7.2 at 37 ℃ for 1 hour. The enzyme reaction was stopped by adding Hepes buffer (pH 7.0) containing 0.4mM KF, 133mMEDTA, 0.1% BSA, and to this buffer, XL 665-labeled anti-histidine antibody and tyrosine phosphorylated monoclonal antibody conjugated with europium cryptate (Eu-K) were added at room temperature and labeled for 1 to 2 hours. The characteristics of the two fluorophores are available from G.Mathis et al, cancer research 1997, 17, pages 3011-3014. The energy transfer between europium cryptate and acceptor XL665 is proportional to the degree of FAK autophosphorylation. Using Packard DisThe coverage plate counter measures the XL-665 specific long-time signal. All content determinations were performed twice and the average of the results of the two content determinations was calculated. The inhibition of FAK autophosphorylation activity of the compounds of the invention is expressed as a percentage inhibition relative to a control, the control activity being determined in the absence of the test compound. The calculated result for% inhibition can be taken as the ratio of 665nm signal/at 620nm signal.

2.KDR

The inhibition of these compounds was determined by the in vitro KDR enzyme in a matrix phosphorylation assay using scintillation technology (96-well plates).

The cytoplasmic domain of the human KDR enzyme was cloned as a GST fusion into the baculovirus expression vector pFastBac. This protein was expressed in SF21 cells and then purified to about 60% homogeneity.

At 20mM MOPS, 10mM MgCl₂、10mM MnCl₂1mM DTT, 2.5mM megta, 10mM b-glycerophosphate, pH7.2, in 10mM MgCl₂、100μMNa₃VO₄KDR kinase activity was measured in the presence of 1mM NaF. Mu.l of the compound was added to 70. mu.l of kinase buffer containing 100ng KDR enzyme at 4 ℃. Mu.l of a mixture containing 2. mu.g of matrix (SH 2-SH3 fragment of PLC expressed as GST fusion protein), 2. mu. Ci³³P[ATP]And a solution of 2. mu.M cold ATP initiated this reaction. After incubation at 37 ℃ for 1 hour, the reaction was stopped by adding 1 volume (100. mu.l) of 200mM EDTA. The culture buffer was removed and the wells were washed three times with 300 μ l PBS. The radioactivity of each well was measured using a Top Count NXT (Packard) radioactivity counter.

Background noise was determined by measuring the radioactivity in four different wells containing radioactive ATP versus substrate only.

In four of the reactors are filled with reactants³³P-[ATP]KDR and PLC matrix), but no compound, control values for total activity were measured.

KDR activity inhibition by a compound of the invention is expressed as a percentage inhibition of control activity determined in the absence of the compound.

Compound SU5614(Calbiochem) (1. mu.M) was included in each plate as an inhibition control.

3.Tie2

The coding sequence of human Tie2 corresponding to the amino acids of the intracellular domain 776-1124 was generated by PCR using cDNA isolated from human placenta as a model. This sequence has been introduced into the baculovirus expression vector pFastBacGT as a GST fusion protein.

The inhibition of these molecules was determined by Tie2 in the presence of GST-Tie2 purified to about 80% homogeneity in the PLC phosphorylation assay. This matrix consists of an SH2-SH3 fragment expressing PLC as a GST fusion protein.

In the presence of 10mM MgCl₂、10mM MnCl₂The kinase activity of Tie2 was measured in 20mM MOPS buffer (pH7.2) containing 1mM DTT and 10mM glycerol phosphate. In FlashPlate 96-well plates kept on ice, a reaction mixture consisting of 70. mu.l of kinase buffer containing 100ng of GST-Tie2 enzyme was placed in each well. Then 10. mu.l of the molecule to be tested are added, diluted in DMSO to a maximum concentration of 10%. For a given concentration, each measurement was performed four times. Add 20. mu.l of a mixture containing 2. mu.g GST-PLC, 2. mu.M cold ATP and 1. mu. Ci³³P[ATP]The solution of (a) initiates the reaction. After incubation at 37 ℃ for 1 hour, the reaction was stopped by adding 1 volume (100. mu.l) of 200mM EDTA. After removal of the culture buffer, the wells were washed three times with 300 μ l PBS. The radioactivity was measured using a Wallac MicroBeta 1450.

Inhibition of Tie2 activity was calculated and expressed as percent inhibition relative to control activity measured in the absence of compound.

As a result: table 1:

Claims

1. A product corresponding to the following formula (I):

in the formula:

2) l is selected from: NH, CO-NH, NH-CO, NH-SO₂，SO₂NH，NH-CH₂，CH₂-NH，CH₂-CO-NH，NH-CO-CH₂，NH-CH₂-CO，CO-CH₂-NH，NH-CO-NH，NH-CS-NH，NH-CO-O，O-CO-NH，CH₂-NH-CO-NH，NH-CO-NH-CH₂，NH-CO-CH₂-CO-NH；

3) Ra is selected from H, alkyl and cycloalkyl;

4) r1 is selected from: H. r, COR, SO₂R, wherein R is selected from H, OR "₄、NR”₅R”₆(C1-C6) alkyl, cycloalkyl, heterocyclyl, substituted heterocyclyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, wherein R "4 is selected from H, phenyl, alkyl, and wherein R" 5 and R "6 are independently selected from H, R OR"₄(C1-C6) alkyl, cycloalkyl, heterocyclyl, substituted heterocyclyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, or R "5 and R" 6 are linked to each other to form a 5-to 8-membered saturated ring containing 0-3 heteroatoms selected from O, S and N;

5) r2 and R5 are independently selected from: H. halogen, R '2, cN, O (R' 2), OC (O) N (R '2) (R' 3), OS (O)₂)(R’2)，N(R’2)(R’3)，N＝C(R’2)(R’3)，N(R’2)C(O)(R’3)，N(R’2)C(O)O(R’3)，N(R’4)C(O)N(R’2)(R’3)，N(R’4)C(S)N(R’2)(R’3)，N(R’2)S(O₂)(R’3)，C(O)(R’2)，C(O)O(R’2)，C(O)N(R’2)(R’3)，C(＝N(R’3))(R’2)，C(＝N(OR’3))(R’2)，S(R’2)，S(O)(R’2)，S(O₂)(R’2)，S(O₂)O(R’2)，S(O₂)N(R’2)(R’3)；

Wherein each R ' 2, R ' 3, R ' 4 is independently selected from H, alkyl, alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl, heterocyclyl, substituted alkyl, substituted alkenyl, substituted alkynyl, substituted aryl, substituted heteroaryl, substituted cycloalkyl, substituted heterocyclyl; wherein each of R '2 and R' 3, when different from H, when present on R2 or R3, are linked to each other to form a ring containing 0 to 3 heteroatoms selected from O, S and N.

2. The product of claim 1, wherein:

1A and Ar are as defined above;

2) r1 is H;

3) l is from: NHCO, NH-CO-NH, NH, NHSO₂，NHCO-CH₂-CONH；

4) Ra is selected from H and methyl;

5) r2 and R5 are as previously defined.

3. The product according to claim 1 or 2, characterized in that Ar-L-a is:

wherein each of X1, X2, X3 and X4 is independently selected from N and C-R '5, wherein R' 5 has the same definition as R2.

4. The product according to claim 3, characterized in that R' 5 is selected from H, F, Cl, methyl, NH₂、OMe、OCF₃And CONH₂。

5. Product according to claim 1 or 2, characterized in that R2 and R5 are independently selected from: H. halogen, R ' 2, OR ' 2, NHR ' 2, NHCOR ' 2, NHCONHR ' 2, NHSO₂R’2。

6. The product of claim 5, wherein R2 is H.

7. The product of claim 5, wherein R5 is H.

8. Product according to any one of claims 1 to 7, characterized in that Ra is H.

9. Product according to any one of claims 1 to 8, characterized in that L-a is chosen from NH-CO-NH-a and NH-SO₂-A。

10. A product according to any one of claims 1 to 9, characterized in that a is selected from phenyl, pyridyl, pyrimidinyl, thienyl, furyl, pyrrolyl, oxazolyl, thiazolyl, isoxazolyl, isothiazolyl, pyrazolyl, imidazolyl, indolyl, indazolyl, benzimidazolyl, benzoxazolyl and benzothiazolyl; optionally substituted.

11. Product according to claim 10, characterized in that a is selected from phenyl, pyrazolyl and isoxazolyl; optionally substituted.

12. The product according to any one of claims 10 to 11, wherein a is substituted with a first substituent selected from the group consisting of alkyl, haloalkyl, alkenyl, alkynyl, aryl, heteroaryl, O-alkyl, O-aryl, O-heteroaryl, S-alkyl, substituted S-alkyl, S-aryl, S-heteroaryl, each of which is optionally substituted with a substituent selected from the group consisting of (C1-C3) alkyl, halogen, O- (C1-C3) alkyl.

13. Product according to any one of claims 10 to 12, characterized in that a is substituted by a second substituent selected from the group consisting of F, Cl, Br, I, OH, SH, SO₃M、COOM、CN、NO₂CON (R8) (R9), N (R8) CO (R9), (C1-C3) alkyl-OH, (C1-C3) alkyl-N (R8) (R9), (C1-C3) alkyl- (R10), (C1-C3) alkyl-COOH, N (R8) (R9); wherein R8 and R9 are independently selected from H, (C1-C3) alkyl, halo (C1-C3) alkyl, (C1-C3) alkyl OH, (C1-C3) alkyl NH₂(C1-C3) alkyl COOM, (C1-C3) alkyl SO₃M; in which R8 and R9 are not both HThey are linked to form a 5-to 7-linked ring containing 0-3 heteroatoms selected from N, O and S; wherein M is H or an alkali metal cation selected from Li, Na and K; and wherein R10 is H or an optionally substituted non-aromatic heterocyclic ring containing 2 to 7 carbon atoms and 1 to 3 heteroatoms selected from N, O and S.

14. Product according to one of claims 10 to 13, characterized in that a is phenyl, pyrazolyl or isoxazolyl which is substituted by halogen, (C1-C4) alkyl, halo (C1-C3) alkyl, O- (C1-C4) alkyl, S- (C1-C4) alkyl, halo O- (C1-C4) alkyl and halo S- (C1-C4) alkyl, and when a is disubstituted, the two substituents can be linked to one another to form a 5-to 7-membered ring which contains 0 to 3 heteroatoms from the group O, N and S.

15. Product according to any one of claims 1 to 14, characterized in that it is:

4- {4- [3- (2-fluoro-5-trifluoromethylphenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide,

1-acetyl-2-amino-4- {4- [3- (2-fluoro-5-trifluoromethylphenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide,

2-formylamino-4- {4- [3- (2-fluoro-5-trifluoromethylphenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide,

2-isobutyrylamino-4- {4- [3- (2-fluoro-5-trifluoromethylphenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide,

2-butyrylamino-4- {4- [3- (2-fluoro-5-trifluoromethylphenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide,

2- (3-cyclopentylpropionylamino) -4- {4- [3- (2-fluoro-5-trifluoromethylphenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide,

2- (cyclopropylcarbonylamino) -4- {4- [3- (2-fluoro-5-trifluoromethylphenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide,

2-tert-pentanoylamino-4- {4- [3- (2-fluoro-5-trifluoromethylphenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide,

2- (2-dimethylaminoacetylamino) -4- {4- [3- (2-fluoro-5-trifluoromethylphenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide,

2-acetylamino-4- {6- [3- (2-fluoro-5-trifluoromethylphenyl) ureido ] pyridin-3-yl } -1H-pyrrole-3-carboxamide,

2- (3-ethylureido) -4- {4- [3- (2-fluoro-5-trifluoromethylphenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide,

2-acetylamino-4- {4- [3- (3-fluoro-5-trifluoromethylphenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide,

2-acetylamino-4- {4- [3- (3-ethylphenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide,

2-acetylamino-4- {4- [3- (4-fluoro-3-trifluoromethylphenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide,

2-acetylamino-4- {4- [3- (3-fluorophenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide,

2-acetylamino-4- {4- [3- (4-fluorophenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide,

2-acetylamino-4- {4- [3- (2-fluorophenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide,

2-acetylamino-4- {4- [3- (4-difluoromethoxyphenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide,

2-acetylamino-4- {4- [3- (3, 4-dimethylphenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide,

2-acetylamino-4- {4- [3- (3, 4-dimethoxyphenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide,

2-acetylamino-4- {4- [3- (2, 5-dimethoxyphenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide,

2-acetylamino-4- [4- (3-phenylureido) phenyl ] -1H-pyrrole-3-carboxamide,

2-acetylamino-4- {4- [3- (2-methoxyphenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide,

2-acetylamino-4- {4- [3- (2-trifluoromethylphenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide,

2-acetylamino-4- [4- (3-o-tolylureido) phenyl ] -1H-pyrrole-3-carboxamide,

2-acetylamino-4- {4- [3- (3-methoxyphenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide,

2-acetylamino-4- {4- [3- (3-trifluoromethylphenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide,

2-acetylamino-4- [4- (3-m-tolylureido) phenyl ] -1H-pyrrole-3-carboxamide

2-acetylamino-4- {4- [3- (4-trifluoromethylphenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide,

2-acetylamino-4- [4- (3-p-tolylureido) phenyl ] -1H-pyrrole-3-carboxamide,

2-acetylamino-4- {4- [3- (4-chloro-3-trifluoromethylphenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide,

2-acetylamino-4- {4- [3- (2-chloro-5-trifluoromethylphenyl) ureido ] phenyl } -lH-pyrrole-3-carboxamide,

2-acetylamino-4- {4- [3- (2-fluoro-3-trifluoromethylphenyl) ureido ] phenyl } -lH-pyrrole-3-carboxamide,

2-acetylamino-4- {4- [3- (3-chloro-4-difluoromethoxyphenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide,

2-acetylamino-4- {4- [3- (3, 5-dimethoxyphenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide,

2-acetylamino-4- {4- [3- (3, 5-dimethylphenyl) ureido ] phenyl } -lH-pyrrole-3-carboxamide,

2-acetylamino-4- {4- [3- (2, 5-dimethylphenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide,

2-acetylamino-4- {4- [3- (2-methoxy-5-methylphenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide,

2-acetylamino-4- {4- [3- (4-methyl-3-trifluoromethylphenyl) ureido ] phenyl } -1H-pyrrole-3-carboxamide,

2-acetylamino-4- [4- (2, 3-dichlorobenzenesulfonylamino) phenyl ] -1H-pyrrole-3-carboxamide.

16. Product according to any one of the preceding claims, characterized in that it is in the form of:

1) achiral, or

2) Racemic, or

3) Is enriched in stereoisomers, or

4) Enriched in enantiomer two;

and in that it is optionally salified.

17. A pharmaceutical composition comprising a product according to any one of the preceding claims in combination with a pharmaceutically acceptable excipient.

18. Use of a product according to any of claims 1 to 16 as an inhibitor of a kinase catalysed reaction.

19. Use of a product according to claim 18, characterized in that the kinase is selected from FAK, KDR and Tie 2.

20. Use of a product according to any one of claims 1 to 16 in the manufacture of a medicament for the treatment of a pathological condition.

21. Use according to claim 20, characterized in that the pathological condition is cancer.