HK1107651B - Inhibitors of the interaction between mdm2 and p53 - Google Patents

Description

Inhibitors of the interaction between MDM2 and P53

Technical Field

The present invention relates to compounds that are useful as inhibitors of the interaction between MDM2 and p53 and compositions comprising the compounds. In addition, the invention provides methods of making the disclosed inhibitors, compositions comprising them, and methods of using them, e.g., as pharmaceuticals.

p53 is a tumor suppressor protein that plays an important role in regulating the balance between cell proliferation and cell growth arrest/apoptosis. Under normal conditions, the half-life of p53 is short, so the concentration of p53 in cells is low. However, in response to cellular DNA damage or cellular stress (e.g., oncogene activation, telomere erosion, hypoxia), the concentration of p53 increases. This increase in p53 concentration results in transcriptional activation of many genes that drive cell growth to stop or enter the apoptotic process. Thus, an important function of p53 is to avoid uncontrolled proliferation of damaged cells and thus to protect the organism from the development of cancer.

MDM2 is a key negative regulator of p53 function. It forms a negative autoregulatory loop by binding to the amino terminal transactivation domain of p53, and thus MDM2 simultaneously inhibits p53 from gaining activated transcriptional capacity and targets p53 for protein degradation. In general, this regulation loop is responsible for maintaining a low concentration of p 53. However, in tumors with wild-type p53, the equilibrium concentration of active p53 can be increased by antagonizing the interaction between MDM2 and p 53. This will result in the restoration of p 53-mediated pro-apoptosis and anti-proliferative effects in such tumor cells.

MDM2 is a cellular proto-oncogene. Over-expression of MDM2 has been observed in cancer regions. MDM2 is overexpressed in various tumors due to gene amplification or increased transcription or translation. The mechanism by which MDM2 amplification promotes tumorigenesis is at least partially related to its interaction with p 53. In cells overexpressing MDM2, the protective function of p53 is hindered, and therefore the cells cannot respond to DNA damage or cellular stress by increasing p53 concentration, resulting in cell growth arrest and/or apoptosis. Thus, cells overexpressing MDM2 are free to continue to proliferate and adopt tumorigenic phenotype after DNA damage and/or cellular stress. In these cases, disruption of the interaction of p53 and MDM2 would release p53 and thus allow the normal signaling of growth arrest and/or apoptosis to occur.

MDM2 may also have different functions in addition to inhibiting p53, e.g. MDM2 has been shown to interact directly with pRb-regulated transcription factor E2F1/DP 1. This interaction is important for the p 53-independent oncogenic activity of MDM 2. The region E2F1 shows significant similarity to the MDM 2-binding p53 region. Since the interactions of MDM2 with p53 and E2F1 are both located at the same binding site of MDM2, it is expected that MDM2/p53 antagonists will not only activate p53 of cells, but also modulate the activity of E2F1, which is generally unregulated in tumor cells.

Also the therapeutic efficacy of recently used DNA damaging agents (chemotherapy and radiotherapy) may be limited by the negative regulation of p53 by MDM 2. Thus, if MDM2 feedback inhibition of p53 is hindered, an increase in functional p53 concentration will increase the therapeutic efficacy of such agents by restoring function to wild-type p53 leading to apoptosis and/or reversing drug resistance associated with p 53. It has been shown that the combination of MDM2 inhibition and in vivo DNA-damage therapy results in a synergistic antitumor effect (Vousden K.H., Cell, Vol. 103, 691-694, 2000).

Thus, disruption of the interaction of MDM2 and p53 provides a method of therapeutic intervention with wild-type p53 in tumors, exhibiting antiproliferative effects even in tumor cells lacking functional p53, and additionally may sensitize tumorigenic cells to chemotherapy and radiotherapy. Background

JP11130750, published on 18.5.1999, describes in particular substituted phenylaminocarbonylindolyl derivatives as 5-HT receptor antagonists.

EP1129074, published at 5/18/2000, describes anthranilic acid amides as inhibitors of Vascular Endothelial Growth Factor Receptors (VEGFR) and useful for the treatment of angiogenic disorders.

EP1317443, published on 3.21.2002, discloses tricyclic tertiary amine derivatives useful as CXCR receptors⁴Or CCR⁵Modulators for the treatment of human immunodeficiency virus and feline immunodeficiency virus.

EP1379329, published 10.10.2002, discloses N- (2-arylethyl) benzylamine as 5-HT₆An antagonist of the receptor. More specifically disclosed are the following:

6-chloro-N- [ [3- (4-pyridinylamino) phenyl ] methyl ] -1H-indole-3-ethylamine,

n- [ [3 (4-pyridinylamino) phenyl ] methyl ] -1H-indole-3-ethylamine, and

5-methoxy-N- [ [3 (4-pyridinylamino) phenyl ] methyl ] -1H-indole-3-ethylamine.

WO 00/15357, published on 23.3.2000, provides piperazine-4-phenyl derivatives as inhibitors of the interaction between MDM2 and p 53. EP 1137418, disclosed on 8.6.2000, provides tricyclic compounds useful for restoring conformational stability to proteins of the p53 family.

WO 03/041715, published on 22.5.2003, describes substituted 1, 4-benzodiazepinesAnd their use as inhibitors of the MDM2-p53 interaction.

WO 03/51359, published at 26.6.2003, provides cis-2, 4, 5-triphenyl-imidazolone, which inhibits the interaction of MDM2 protein with p 53-like peptides and has antiproliferative activity.

WO 04/05278, disclosed on 15/1/2004, discloses bisaryl sulfonamide compounds that bind MDM2 and are useful for the treatment of cancer.

There is a continuing need for effective and potent small molecules that inhibit the interaction between MDM2 and p 53.

The compounds of the present invention differ from the prior art in structure, their pharmacological activity and/or pharmacological potency.

Description of the invention

The present invention provides compounds, compositions and methods for inhibiting the interaction between MDM2 and p53 for the treatment of cancer. In addition, the compounds and compositions of the present invention may be used to enhance the effectiveness of chemotherapy and radiotherapy.

The present invention relates to compounds of formula (I).

A compound of the formula (I),

an N-oxide form, an addition salt or a stereochemically isomeric form thereof, wherein:

m is 0, 1 or 2, and when m is 0, it means a direct bond;

n is 0, 1, 2 or 3, and when n is 0, it means a direct bond;

p is 0 or 1, and when p is 0, it means a direct bond;

s is 0 or 1, and when s is 0, it means a direct bond;

t is 0 or 1, and when t is 0, it means a direct bond;

x is C (═ O) or CHR⁸(ii) a Wherein

R⁸Is hydrogen, C_1-6Alkyl radical, C_3-7Cycloalkyl, -C (═ O) -NR¹⁷R¹⁸Hydroxycarbonyl radical, aryl radical C_1-6Alkoxycarbonyl, heteroaryl, heteroarylcarbonyl, heteroaryl C_1-6Alkoxycarbonyl, piperazinylcarbonyl, pyrrolidinyl, piperidinylcarbonyl, C_1-6Alkoxycarbonyl, C substituted by a substituent selected from the group consisting of hydroxy, amino, aryl and heteroaryl_1-6An alkyl group; c substituted by a substituent selected from the group consisting of hydroxy, amino, aryl and heteroaryl_3-7A cycloalkyl group; by hydroxy, hydroxy C_1-6Alkyl, hydroxy C_1-6Alkoxy radical C_1-6An alkyl-substituted piperazinecarbonyl group; by hydroxy radicals C_1-6Alkyl-substituted pyrrolidinyl; or by one or two radicals selected from hydroxy, C_1-6Alkyl, hydroxy C_1-6Alkyl radical, C_1-6Alkoxy radical C_1-6Alkyl radical, C_1-6Alkyl (dihydroxy) C_1-6Alkyl or C_1-6Alkoxy (hydroxy) C_{1 -6}Piperidinyl carbonyls substituted with alkyl substituentsA group;

R¹⁷and R¹⁸Each independently selected from hydrogen and C_1-6Alkyl, di (C)_1-6Alkyl) amino C_1-6Alkyl, aryl C_1-6Alkyl radical, C_1-6Alkoxy radical C_1-6Alkyl, hydroxy C_1-6Alkyl, hydroxy C_1-6Alkyl radical (C)_1-6Alkyl) or hydroxy C_1-6Alkyl (aryl C)_{1-6 alkyl}Radical);

is-CR⁹C and the dotted line is a bond, -C (O) -CH <, -C (O) -N <, -CHR⁹-CH < or-CHR⁹-N <; wherein

Each R⁹Independently is hydrogen or C_1-6An alkyl group;

R¹is hydrogen, aryl, heteroaryl, C_1-6Alkoxycarbonyl group, C_1-12Alkyl or substituted by one or two groups independently selected from hydroxy, aryl, heteroaryl, amino, C_1-6Alkoxy, mono-or di (C)_{1 -6}Alkyl) amino, morpholinyl, piperidinyl, pyrrolidinyl, piperazinyl, C_1-6Alkyl piperazinyl, aryl C_1-6Alkyl piperazinyl, heteroaryl C_1-6Alkyl piperazinyl, C_3-7Cycloalkyl piperazinyl and C_3-7Cycloalkyl radical C_1-6C substituted by substituents of alkylpiperazino radicals_1-12An alkyl group;

R²is hydrogen, halogen, C_1-6Alkyl radical, C_1-6Alkoxy, aryl C_1-6Alkoxy, heteroaryl C_1-6Alkoxy, phenylthio, hydroxy C_1-6Alkylcarbonyl, C substituted by a substituent selected from amino, aryl and heteroaryl_1-6An alkyl group; or C substituted by a substituent selected from the group consisting of amino, aryl and heteroaryl_3-7A cycloalkyl group;

R³is hydrogen, C_1-6Alkyl, heteroarylBase, C_3-7Cycloalkyl, C substituted by a substituent selected from the group consisting of hydroxy, amino, aryl and heteroaryl_1-6An alkyl group; or C substituted by a substituent selected from the group consisting of hydroxy, amino, aryl and heteroaryl_3-7A cycloalkyl group;

R⁴and R⁵Each independently of the others is hydrogen, halogen, C_1-6Alkyl, polyhalo C_1-6Alkyl, cyano C_1-6Alkyl, hydroxy, amino or C_1-6An alkoxy group; or

R⁴And R⁵Together optionally forming a divalent group selected from methylenedioxy or ethylenedioxy;

R⁶is hydrogen, C_1-6Alkoxycarbonyl or C_1-6An alkyl group;

when p is 1, then R⁷Is hydrogen, aryl C_1-6Alkyl, hydroxy or heteroaryl C_1-6An alkyl group;

z is a group selected from:

wherein

Each R¹⁰Or R¹¹Each independently selected from hydrogen, halogen, hydroxy, amino, C_1-6Alkyl, nitro, polyhalo C_1-6Alkyl, cyano C_1-6Alkyl, tetrazole C_1-6Alkyl, aryl, heteroaryl, aryl C_1-6Alkyl, heteroaryl C_1-6Alkyl, aryl (hydroxy) C_1-6Alkyl, heteroaryl (hydroxy) C_1-6Alkyl, arylcarbonyl, heteroarylcarbonyl, C_1-6Alkylcarbonyl, aryl C_1-6Alkylcarbonyl, heteroaryl C_1-6Alkylcarbonyl group, C_1-6Alkoxy radical, C_3-7Cycloalkyl carbonyl group, C_3-7Cycloalkyl (hydroxy) C_1-6Alkyl, aryl C_1-6Alkoxy radical C_1-6Alkyl radical, C_1-6Alkoxy radical C_1-6Alkoxy radical C_1-6Alkyl radical, C_1-6Alkylcarbonyloxy C_1-6Alkyl radical, C_1-6Alkoxycarbonyl radical C_1-6Alkoxy radical C_1-6Alkyl, hydroxy C_1-6Alkoxy radical C_1-6Alkyl radical, C_1-6Alkoxycarbonyl radical C_2-6Alkenyl radical, C_1-6Alkoxy radical C_1-6Alkyl radical, C_1-6Alkoxycarbonyl group, C_1-6Alkylcarbonyloxy, aminocarbonyl, hydroxy C_1-6Alkyl, amino C_1-6Alkyl, hydroxycarbonyl C_1-6Alkyl and- (CH)₂)_v-(C(＝O)_r)-(CHR¹⁹)_u-NR¹³R¹⁴(ii) a Wherein

v is 0, 1, 2, 3, 4, 5 or 6, and when v is 0, it means a direct bond;

r is 0 or 1, and when r is 0, it means a direct bond;

u is 0, 1, 2, 3, 4, 5 or 6, and when u is 0, it means a direct bond;

R¹⁹is hydrogen or C_1-6An alkyl group;

R¹²is hydrogen, C_1-6Alkyl radical, C_3-7Cycloalkyl, selected from hydroxy, amino, C_1-6C substituted by alkoxy and aryl substituents_1-6An alkyl group; or selected from hydroxy, amino, aryl and C_1-6C substituted by substituents of alkoxy_3-7A cycloalkyl group;

R¹³and R¹⁴Each independently selected from hydrogen and C_1-12Alkyl radical, C_1-6Alkylcarbonyl group, C_1-6Alkylsulfonyl, aryl C_1-6Alkylcarbonyl group, C_3-7Cycloalkyl radical, C_3-7Cycloalkyl carbonyl, - (CH)₂)_k-NR¹⁵R¹⁶Selected from hydroxy, hydroxycarbonyl, cyano, C_1-6Alkoxycarbonyl group, C_1-6C substituted by substituents of alkoxy, aryl or heteroaryl_1-12An alkyl group; or selected from hydroxy, C_1-6Alkoxy, aryl, amino, aryl C_1-6Alkyl, heteroaryl or heteroaryl C_1-6C substituted by substituents of alkyl groups_3-7A cycloalkyl group; or

R¹³And R¹⁴Together with the nitrogen to which they are attached may optionally form morpholinyl, piperidinyl, pyrrolidinyl, piperazinyl or be selected from C_1-6Alkyl, aryl C_1-6Alkyl, aryl C_1-6Alkoxycarbonyl, heteroaryl C_1-6Alkyl radical, C_3-7Cycloalkyl and C_3-7Cycloalkyl radical C_1-6Piperazinyl substituted with a substituent for alkyl; wherein

k is 0, 1, 2, 3, 4, 5 or 6, and when k is 0, it means a direct bond;

R¹⁵and R¹⁶Each independently selected from hydrogen and C_1-6Alkyl, aryl C_1-6Alkoxycarbonyl group, C_3-7Cycloalkyl, selected from hydroxy, C_1-6C substituted by substituents of alkoxy, aryl and heteroaryl_1-12An alkyl group; and is selected from hydroxy, C_1-6Alkoxy, aryl C_1-6Alkyl, heteroaryl and heteroaryl C_1-6C substituted by substituents of alkyl groups_3-7A cycloalkyl group; or

R¹⁵And R¹⁶Together with the nitrogen to which they are attached may optionally form morpholinyl, piperazinyl or be selected from C_1-6Piperazinyl substituted with alkoxycarbonyl;

aryl is phenyl or naphthyl;

each phenyl or naphthyl group may be optionally substituted with one, two or three substituents each independently selected from halogen, hydroxy, C_1-6Alkyl, amino, polyhalo C_1-6Alkyl and C_1-6An alkoxy group; and

each phenyl or naphthyl group may be optionally substituted with a divalent group selected from methylenedioxy or ethylenedioxy;

heteroaryl is pyridyl, indolyl, quinolinyl, imidazolyl, furyl, thienyl, oxadiazolyl, tetrazolyl, benzofuryl or tetrahydrofuranyl;

each pyridyl, indolyl, quinolinyl, imidazolyl, furyl, thienyl, oxadiazolyl, tetrazolyl, benzofuryl or tetrahydrofuranyl group may be optionally substituted with one, two or three substituents each independently selected from halogen, hydroxy, C_1-6Alkyl, amino, polyhalo C_1-6Alkyl, aryl C_1-6Alkyl or C_1-6An alkoxy group; and

each pyridyl, indolyl, quinolinyl, imidazolyl, furyl, thienyl, benzofuryl or tetrahydrofuranyl group may be optionally substituted with a divalent group selected from methylenedioxy or ethylenedioxy;

with the following conditions:

when m is 1; by removal of R from the benzene ring²The outer substituents are in the meta position; s is 0; and t is 0, then

Z is a group selected from (a-1), (a-3), (a-4), (a-5), (a-6), (a-7), (a-8) or (a-9).

The compounds of formula (I) also exist in their tautomeric form. Such forms, although not explicitly shown in the above formula, are intended to be included within the scope of the present invention.

Many of the terms used in the above definitions and hereafter are explained below. These terms are sometimes used in the original or in compound terms.

When used in the above definitions and hereafter, halo broadly refers to fluoro, chloro, bromo and iodo; c_1-6Alkyl defines straight and branched chain saturated hydrocarbon groups having from 1 to 6 carbon atoms, such as methyl, ethyl, propyl, butyl, pentyl, hexyl, 1-methylethyl, 2-methylpropyl, 2-methyl-butyl, 2-methylpentyl, and the like; c_1-6Alkanediyl defines divalent straight and branched chain saturated hydrocarbon radicals having from 1 to 6 carbon atoms, e.g. sulfinoMethyl, 1, 2-ethanediyl, 1, 3-propanediyl, 1, 4-butanediyl, 1, 5-pentanediyl, 1, 6-hexanediyl and branched isomers thereof, such as 2-methylpentanediyl, 3-methylpentanediyl, 2-dimethylbutanediyl, 2, 3-dimethylbutanediyl and the like; c_{1- 12}The alkyl group comprising C_1-6Alkyl groups and their higher homologues having from 7 to 12 carbon atoms, such as heptyl, octyl, nonyl, decyl, undecyl, and dodecyl; hydroxy radical C_1-6Alkyl defines a hydroxy substituent on straight and branched chain saturated hydrocarbon groups having 1 to 6 carbon atoms; trihalomethyl defines methyl containing three identical or different halogen substituents, for example trifluoromethyl; c_{2 -6}Alkenyl defines straight and branched chain hydrocarbon groups containing one double bond and having 2 to 6 carbon atoms, such as vinyl, 2-propenyl, 3-butenyl, 2-pentenyl, 3-methyl-2-butenyl, and the like; c_3-7Alkynyl defines straight and branched chain hydrocarbon radicals containing one triple bond and having 3 to 6 carbon atoms, such as 2-propynyl, 3-butynyl, 2-pentynyl, 3-hexynyl, and the like; c_3-7Cycloalkyl groups include cyclic hydrocarbon groups having 3 to 10 carbons, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cycloheptyl, and the like.

The term "addition salts" comprises the salts which the compounds of formula (I) can form with organic or inorganic bases, such as amines, alkali metal bases and alkaline earth metal bases or quaternary ammonium bases, or with organic or inorganic acids, such as mineral acids, sulfonic acids, carboxylic acids or phosphorus-containing acids.

The term "addition salts" further comprises the pharmaceutically acceptable salts, metal complexes, and solvates and salts thereof which the compounds of formula (I) are capable of forming.

The term "pharmaceutically acceptable salt" means a pharmaceutically acceptable acid or base addition salt. The pharmaceutically acceptable acid or base addition salts as mentioned hereinbefore are meant to comprise the therapeutically active non-toxic acid and non-toxic base addition salt forms which the compounds of formula (I) are able to form. Compounds of formula (I) having basic properties may be converted into their pharmaceutically acceptable acid addition salts by treating the base form with an appropriate acid. Suitable acids include, for example, inorganic acids, such as hydrohalic acids, e.g., hydrochloric or hydrobromic acid; sulfuric acid; nitric acid; phosphoric acid and the like; or organic acids such as acetic, propionic, glycolic, lactic, pyruvic, oxalic, malonic, succinic (i.e., succinic), maleic, fumaric, malic, tartaric, citric, methanesulfonic, ethanesulfonic, benzenesulfonic, p-toluenesulfonic, cyclamic, salicylic, p-aminosalicylic, pamoic and the like acids.

The compounds of formula (I) having acidic properties may be converted into their pharmaceutically acceptable base addition salts by treating the acid form with a suitable organic or inorganic base. Suitable base salt forms include, for example, ammonium salts, alkali and alkaline earth metal salts, such as lithium, sodium, potassium, magnesium, calcium salts and the like, salts with organic bases, such as benzathine (benzathine), N-methyl-D-glucosamine, hydrabamine (hydrabamine) salts, and salts with amino acids, such as arginine, lysine and the like.

The term acid or base addition salt also encompasses hydrates and solvent addition forms which the compounds of formula (I) are able to form. Examples of such forms are, for example, hydrates, alcoholates and the like.

The term "metal complex" means a complex formed between a compound of formula (I) and one or more organic or inorganic metal salts. Examples of such organic or inorganic salts include halides, nitrates, sulfates, phosphates, acetates, trifluoroacetates, trichloroacetates, propionates, tartrates, sulfonates (e.g., methylsulfonates, 4-methylphenyl sulfonates), salicylates, benzoates and salts of metals of the second main group of the periodic table (e.g., magnesium or calcium salts), of the third or fourth main group (e.g., aluminum, tin, lead) and of the first to eighth transition groups of the periodic table (e.g., chromium, manganese, iron, cobalt, nickel, copper, zinc, etc.).

As used hereinbefore, the term "stereochemically isomeric forms of a compound of formula (I)" is defined as all possible compounds which the compound of formula (I) may possess, consisting of the same atoms bonded by bonds of the same order, but having different three-dimensional structures which are not interconvertible. Unless otherwise mentioned or indicated, the chemical designation of a compound encompasses the mixture of all possible stereochemically isomeric forms which the compound may possess. The mixture may contain all diastereomers and/or optical isomers of the basic molecular structure of the compound. All stereochemically isomeric forms of the compounds of formula (I) in pure form or in admixture with each other are intended to be embraced within the scope of the present invention.

The N-oxide forms of the compounds of formula (I) are meant to comprise those compounds of formula (I) wherein one or several nitrogen atoms are oxidized to the so-called N-oxides, in particular those N-oxides wherein one or more piperidine-, piperazine-or pyridazinyl-nitrogens are N-oxidized.

The term "compound of formula (I)" when used hereinafter is meant to also include the N-oxide form, the pharmaceutically acceptable acid or base addition salts and all stereoisomeric forms.

A first group of interesting compounds consists of those compounds of formula (I) wherein one or more of the following restrictions apply:

a) x is C (═ O) or CHR⁸(ii) a Wherein

R⁸Is hydrogen, C_1-6Alkyl radical, C_3-7Cycloalkyl, aminocarbonyl, mono-or di (C)_1-6Alkyl) aminocarbonyl, hydroxycarbonyl, arylC_1-6Alkoxycarbonyl, heteroaryl C_1-6Alkoxycarbonyl group, C_1-6Alkoxycarbonyl, C substituted by a substituent selected from the group consisting of hydroxy, amino, aryl and heteroaryl_1-6Alkyl or C substituted by a substituent selected from hydroxy, amino, aryl and heteroaryl_{3 -7}A cycloalkyl group;

b)R¹is hydrogen, aryl, heteroaryl, C_1-12Alkyl or substituted by one or two groups independently selected from hydroxy, aryl, heteroaryl, amino, C_1-6Alkoxy, mono-or di (C)_1-6Alkyl) amino, morpholinyl, piperidinyl, pyrrolidinyl, piperazinyl, piperidinyl, pyrrolidinyl, piperidinyl, and piperazinyl,C_1-6Alkyl piperazinyl, aryl C_1-6Alkyl piperazinyl, heteroaryl C_1-6Alkyl piperazinyl, C_3-7Cycloalkyl piperazinyl and C_3-7Cycloalkyl radical C_{1 -6}C substituted by substituents of alkylpiperazino radicals_1-12An alkyl group;

c)R³is hydrogen, C_1-6Alkyl radical, C_3-7Cycloalkyl, C substituted by a substituent selected from the group consisting of hydroxy, amino, aryl and heteroaryl_1-6An alkyl group; or C substituted by a substituent selected from the group consisting of hydroxy, amino, aryl and heteroaryl_3-7A cycloalkyl group;

d)R⁴and R⁵Each independently of the others is hydrogen, halogen, C_1-6Alkyl, polyhalo C_1-6Alkyl, hydroxy, amino or C_1-6An alkoxy group;

e)R⁴and R⁵Together optionally forming a divalent group selected from methylenedioxy or ethylenedioxy;

f)R⁶is hydrogen or C_1-6An alkyl group;

g) when p is 1, then R⁷Is hydrogen, aryl C_1-6Alkyl or heteroaryl C_1-6An alkyl group;

h) z is a group selected from (a-1), (a-2), (a-3), (a-4), (a-5) and (a-6);

i) each R¹⁰Or R¹¹Each independently selected from hydrogen, hydroxy, amino, C_1-6Alkyl, nitro, polyhalo C_1-6Alkyl, cyano C_1-6Alkyl, tetrazole C_1-6Alkyl, aryl, heteroaryl, aryl C_1-6Alkyl, heteroaryl C_1-6Alkyl, aryl (hydroxy) C_1-6Alkyl, heteroaryl (hydroxy) C_{1 -6}Alkyl, arylcarbonyl, heteroarylcarbonyl, arylC_1-6Alkylcarbonyl, heteroaryl C_1-6Alkylcarbonyl group, C_1-6Alkoxy radical, C_1-6Alkoxy radical C_1-6Alkyl radical, C_1-6Alkoxycarbonyl group, C_1-6Alkylcarbonyloxy, aminocarbonyl, hydroxy C_1-6Alkyl, amino C_1-6Alkyl, hydroxycarbonyl C_1-6Alkyl and- (CH)₂)_v-(C(＝O)_r)-(CH₂)_u-NR¹³R¹⁴；

j)R¹³And R¹⁴Each independently selected from hydrogen and C_1-12Alkyl radical, C_3-7Cycloalkyl, - (CH)₂)_k-NR¹⁵R¹⁶Selected from hydroxy, C_1-6C substituted by substituents of alkoxy, aryl and heteroaryl_1-12An alkyl group; or selected from hydroxy, C_1-6Alkoxy, aryl C_1-6Alkyl, heteroaryl and heteroaryl C_1-6C substituted by substituents of alkyl groups_3-7A cycloalkyl group;

k)R¹³and R¹⁴Together with the nitrogen to which they are attached may optionally form morpholinyl, piperidinyl, pyrrolidinyl, piperazinyl or be selected from C_1-6Alkyl, aryl C_1-6Alkyl, heteroaryl C_1-6Alkyl radical, C_3-7Cycloalkyl and C_3-7Cycloalkyl radical C_1-6Piperazinyl substituted with a substituent for alkyl;

l)R¹⁵and R¹⁶Each independently selected from hydrogen and C_1-6Alkyl radical, C_3-7Cycloalkyl, selected from hydroxy, C_1-6C substituted by substituents of alkoxy, aryl and heteroaryl_1-12An alkyl group; and is selected from hydroxy, C_1-6Alkoxy, aryl C_1-6Alkyl, heteroaryl and heteroaryl C_1-6C substituted by substituents of alkyl groups_3-7A cycloalkyl group;

m) heteroaryl is pyridyl, indolyl, quinolinyl, imidazolyl, furyl, thienyl, benzofuryl or tetrahydrofuranyl; and each pyridyl, indolyl, quinolinyl, imidazolyl, furyl, thienyl, benzofuryl or tetrahydrofuranyl group may optionally be substituted with one, two or three substituents each independently selected from halogen, hydroxy, C_{1 -6}Alkyl, amino, polyhalo C_1-6Alkyl and C_1-6An alkoxy group; and

n) each pyridyl, indolyl, quinolinyl, imidazolyl, furyl, thienyl, benzofuryl or tetrahydrofuranyl group may be optionally substituted by a divalent group selected from methylenedioxy or ethylenedioxy;

a second group of interesting compounds consists of those compounds of formula (I) wherein one or more of the following restrictions apply:

a) n is 0, 1 or 2;

b) p is 0;

c)R⁸is hydrogen, aminocarbonyl, aryl C_1-6Alkoxycarbonyl or C substituted by hydroxy_1-6An alkyl group;

d)is-CR⁹Is < C or-CHR⁹-CH＜；

e)R¹Is hydrogen, C_1-12Alkyl or C substituted by heteroaryl_1-12An alkyl group;

f)R²is hydrogen, halogen, C_1-6Alkyl radical, C_1-6Alkoxy, aryl C_1-6Alkoxy or thiophenyl;

g)R³is hydrogen or C_1-6An alkyl group;

h)R⁴and R⁵Each independently is hydrogen, halogen or C_1-6An alkoxy group;

i) z is a group selected from (a-1), (a-2), (a-3), (a-4) or (a-6);

j) each R¹⁰Or R¹¹Each independently selected from hydrogen, hydroxy, amino, C_1-6Alkyl, nitro, polyhalo C_1-6Alkyl, cyano, aryl C_1-6Alkyl, aryl(hydroxy) C_1-6Alkyl, arylcarbonyl, C_1-6Alkoxy radical, C_1-6Alkoxy radical C_1-6Alkyl radical, C_1-6Alkoxycarbonyl, aminocarbonyl, hydroxy C_1-6Alkyl, amino C_1-6Alkyl, hydroxycarbonyl and- (CH)₂)_v-(C(＝O)_r)-(CH₂)_u-NR¹³R¹⁴；

k) v is 0 or 1;

l) r is 0 or 1;

m) u is 0;

n)R¹³and R¹⁴Each independently selected from hydrogen and C_1-6Alkyl, - (CH)₂)_k-NR¹⁵R¹⁶And C substituted by hydroxy_1-12An alkyl group;

o)R¹³and R¹⁴Together with the nitrogen to which they are attached may form a pyrrolidinyl group;

p) k is 2;

q)R¹⁵and R¹⁶Each independently is C_1-6An alkyl group;

r) aryl is phenyl or phenyl substituted by halogen; and

s) heteroaryl is pyridyl or indolyl.

A third group of interesting compounds consists of those compounds of formula (I) wherein one or more of the following restrictions apply:

a) m is 0 or 2;

b) n is 0, 2 or 3;

c) p is 1;

d) s is 1;

e) t is 1;

f) x is C (═ O);

g)is-C (═ O) -CH <, -C (═ O) -N <, -CHR⁹-CH < or-CHR⁹-N＜；

h)R¹Is aryl, heteroaryl, C_1-6Alkoxycarbonyl group, C_1-12Alkyl or substituted by one or two groups independently selected from hydroxy, aryl, heteroaryl, amino, C_1-6Alkoxy, mono-or di (C)_1-6Alkyl) amino, morpholinyl, piperidinyl, pyrrolidinyl, piperazinyl, C_1-6Alkyl piperazinyl, aryl C_1-6Alkyl piperazinyl, heteroaryl C_1-6Alkyl piperazinyl, C_3-7Cycloalkyl piperazinyl and C_3-7Cycloalkyl radical C_1-6C substituted by substituents of alkylpiperazino radicals_1-12An alkyl group;

i)R²is halogen, C_1-6Alkyl radical, C_1-6Alkoxy, aryl C_1-6Alkoxy, heteroaryl C_1-6Alkoxy, phenylthio, hydroxy C_1-6Alkylcarbonyl, C substituted by a substituent selected from amino, aryl and heteroaryl_1-6An alkyl group; or C substituted by a substituent selected from the group consisting of amino, aryl and heteroaryl_3-7A cycloalkyl group;

j)R³is C_1-6Alkyl radical, C_3-7Cycloalkyl, C substituted by a substituent selected from the group consisting of hydroxy, amino, aryl and heteroaryl_1-6An alkyl group; or C substituted by a substituent selected from the group consisting of hydroxy, amino, aryl and heteroaryl_3-7A cycloalkyl group;

k)R⁴and R⁵Each independently is C_1-6Alkyl, polyhalo C_1-6Alkyl, cyano C_1-6Alkyl, hydroxy or amino;

l)R⁴and R⁵Together optionally forming a divalent group selected from methylenedioxy or ethylenedioxy;

m)R⁶is C_1-6Alkoxycarbonyl or C_1-6An alkyl group;

n)R⁷is hydrogen, aryl C_1-6Alkyl, hydroxy or heteroaryl C_1-6An alkyl group; and

o) Z is a group selected from (a-1), (a-3), (a-4), (a-5), (a-6), (a-7), (a-8) or (a-9).

A fourth group of interesting compounds consists of those compounds of formula (I) wherein one or more of the following restrictions apply:

a)R⁸is hydrogen, -C (═ O) -NR¹⁷R¹⁸Aryl radical C_1-6Alkoxycarbonyl, hydroxy-substituted C_1-6Alkyl, by hydroxy, hydroxy C_1-6Alkyl, hydroxy C_1-6Alkoxy radical C_1-6Alkyl-substituted piperazinecarbonyl, substituted by hydroxy C_1-6Pyrrolidinyl substituted by alkyl or by one or two selected from hydroxy, C_1-6Alkyl, hydroxy C_1-6Alkyl radical, C_1-6Alkoxy radical C_1-6Alkyl radical, C_1-6Alkyl (dihydroxy) C_1-6Alkyl or C_1-6Alkoxy (hydroxy) C_1-6Piperidinylcarbonyl substituted with a substituent for alkyl;

b)R¹⁷and R¹⁸Each independently selected from hydrogen and C_1-6Alkyl, di (C)_1-6Alkyl) amino C_{1 -6}Alkyl, aryl C_1-6Alkyl radical, C_1-6Alkoxy radical C_1-6Alkyl or hydroxy C_1-6An alkyl group;

c)is-CR⁹C < and the dotted line is a bond, -CHR⁹-CH < or-CHR⁹-N＜；

d)R¹Is hydrogen, heteroaryl, C_1-6Alkoxycarbonyl group, C_1-12Alkyl or C substituted by heteroaryl_1-12An alkyl group;

e)R²is hydrogen, halogen, C_1-6Alkyl radical, C_1-6Alkoxy, aryl C_1-6Alkoxy or thiophenyl;

f)R³is hydrogen, C_1-6Alkyl or heteroaryl;

g)R⁴and R⁵Each independently of the others is hydrogen, halogen, C_1-6Alkyl, cyano C_1-6Alkyl, hydroxy or C_1-6An alkoxy group;

h) when p is 1, then R⁷Is aryl C_1-6Alkyl or hydroxy;

i) z is a group selected from (a-1), (a-2), (a-3), (a-4), (a-5), (a-6), (a-8), (a-9), (a-10) and (a-11);

j) each R¹⁰Or R¹¹Each independently selected from hydrogen, halogen, hydroxy, amino, C_1-6Alkyl, nitro, polyhalo C_1-6Alkyl, cyano C_1-6Alkyl, tetrazole C_1-6Alkyl, aryl, heteroaryl C_1-6Alkyl, aryl (hydroxy) C_1-6Alkyl, arylcarbonyl, C_1-6Alkylcarbonyl group, C_3-7Cycloalkyl carbonyl group, C_3-7Cycloalkyl (hydroxy) C_1-6Alkyl, aryl C_1-6Alkoxy radical C_{1 -6}Alkyl radical, C_1-6Alkoxy radical C_1-6Alkoxy radical C_1-6Alkyl radical, C_1-6Alkylcarbonyloxy C_1-6Alkyl radical, C_1-6Alkoxycarbonyl radical C_1-6Alkoxy radical C_1-6Alkyl, hydroxy C_1-6Alkoxy radical C_1-6Alkyl radical, C_1-6Alkoxycarbonyl radical C_2-6Alkenyl radical, C_1-6Alkoxy radical C_1-6Alkyl radical, C_1-6Alkoxycarbonyl, aminocarbonyl, hydroxy C_1-6Alkyl, amino C_1-6Alkyl, hydroxycarbonyl C_1-6Alkyl and- (CH)₂)_v-(C(＝O)_r)-(CHR¹⁹)_u-NR¹³R¹⁴；

k) v is 0 or 1;

l) u is 0 or 1

m)R¹²Is hydrogen or C_1-6An alkyl group;

n)R¹³and R¹⁴Each independently selected from hydrogen and C_1-12Alkyl radical, C_1-6Alkylcarbonyl group, C_{1- 6}Alkylsulfonyl, aryl C_1-6Alkylcarbonyl group, C_3-7Cycloalkyl carbonyl, - (CH)₂)_k-NR¹⁵R¹⁶Selected from hydroxy, hydroxycarbonyl, cyano, C_1-6C substituted by substituents of alkoxycarbonyl or aryl groups_1-12An alkyl group;

o)R¹³and R¹⁴Together with the nitrogen to which they are attached may optionally form morpholinyl, pyrrolidinyl, piperazinyl or be selected from C_1-6Alkyl or aryl radicals C_1-6Piperazinyl substituted with a substituent of alkoxycarbonyl;

p) k is 2;

q)R¹⁵and R¹⁶Each independently selected from hydrogen and C_1-6Alkyl or aryl radicals C_1-6An alkoxycarbonyl group;

r)R¹⁵and R¹⁶Together with the nitrogen to which they are attached may optionally form morpholinyl, piperazinyl or by C_1-6Alkoxycarbonyl-substituted piperazinyl;

s) aryl is phenyl or phenyl substituted by halogen;

t) heteroaryl is pyridyl, indolyl, oxadiazolyl or tetrazolyl; and

u) each pyridyl, indolyl, oxadiazolyl or tetrazolyl group may optionally be substituted by one group selected from C_1-6Alkyl, aryl or aryl C_1-6Alkyl substituents.

A fifth group of interesting compounds consists of those compounds of formula (I) wherein one or more of the following restrictions apply:

a) m is 0;

b) n is 1;

c) p is 0;

d) s is 0;

e) t is 0;

f) x is CHR⁸；

g)R⁸Is hydrogen;

h)is-CR⁹＝C＜；

i) Each R⁹Is hydrogen;

j)R¹is hydrogen;

k)R²is hydrogen or C_1-6An alkoxy group;

l)R³is hydrogen;

m)R⁴and R⁵Each independently is hydrogen, C_1-6Alkyl or C_1-6An alkoxy group;

n)R⁶is hydrogen;

o) Z is a group selected from (a-1), (a-2), (a-3) or (a-4);

p)R¹⁰or R¹¹Each independently selected from hydrogen, hydroxy or hydroxy C_1-6An alkyl group.

A preferred group of compounds consists of those of formula (I) or any subgroup thereof, wherein

R⁸Is hydrogen, -C (═ O) -NR¹⁷R¹⁸Aryl radical C_1-6Alkoxycarbonyl, hydroxy-substituted C_1-6Alkyl, by hydroxy, hydroxy C_1-6Alkyl, hydroxy C_1-6Alkoxy radical C_1-6Alkyl-substituted piperazinecarbonyl, substituted by hydroxy C_1-6Pyrrolidinyl substituted by alkyl or by one or two selected from hydroxy, C_1-6Alkyl, aryl, heteroaryl, and heteroaryl,Hydroxy radical C_1-6Alkyl radical, C_1-6Alkoxy radical C_1-6Alkyl radical, C_1-6Alkyl (dihydroxy) C_1-6Alkyl or C_1-6Alkoxy (hydroxy) C_1-6Piperidinylcarbonyl substituted with a substituent for alkyl; r¹⁷And R¹⁸Each independently selected from hydrogen and C_1-6Alkyl, di (C)_1-6Alkyl) amino C_{1 -6}Alkyl, aryl C_1-6Alkyl radical, C_1-6Alkoxy radical C_1-6Alkyl or hydroxy C_1-6An alkyl group;is-CR⁹C < and the dotted line is a bond, -CHR⁹-CH < or-CHR⁹-N＜；R¹Is hydrogen, heteroaryl, C_1-6Alkoxycarbonyl group, C_1-12Alkyl or C substituted by heteroaryl_1-12An alkyl group; r²Is hydrogen, halogen, C_1-6Alkyl radical, C_1-6Alkoxy, aryl C_1-6Alkoxy or thiophenyl; r³Is hydrogen, C_1-6Alkyl or heteroaryl; r⁴And R⁵Each independently of the others is hydrogen, halogen, C_1-6Alkyl, cyano C_1-6Alkyl, hydroxy or C_1-6An alkoxy group; when p is 1, then R⁷Is aryl C_1-6Alkyl or hydroxy; z is a group selected from (a-1), (a-2), (a-3), (a-4), (a-5), (a-6), (a-8), (a-9), (a-10) and (a-11); each R¹⁰Or R¹¹Each independently selected from hydrogen, halogen, hydroxy, amino, C_1-6Alkyl, nitro, polyhalo C_1-6Alkyl, cyano C_1-6Alkyl, tetrazole C_1-6Alkyl, aryl, heteroaryl C_1-6Alkyl, aryl (hydroxy) C_1-6Alkyl, arylcarbonyl, C_1-6Alkylcarbonyl group, C_3-7Cycloalkyl carbonyl group, C_3-7Cycloalkyl (hydroxy) C_1-6Alkyl, aryl C_1-6Alkoxy radical C_1-6Alkyl radical, C_1-6Alkoxy radical C_1-6Alkoxy radical C_1-6Alkyl radical, C_1-6Alkylcarbonyloxy C_1-6Alkyl radical, C_1-6Alkoxycarbonyl radical C_1-6Alkoxy radical C_1-6Alkyl, hydroxy C_1-6Alkoxy radical C_1-6Alkyl radical, C_1-6Alkoxycarbonyl radical C_2-6Alkenyl radical, C_1-6Alkoxy radical C_1-6Alkyl radical, C_1-6Alkoxycarbonyl, aminocarbonyl, hydroxy C_1-6Alkyl, amino C_1-6Alkyl, hydroxycarbonyl C_1-6Alkyl and- (CH)₂)_v-(C(＝O)_r)-(CHR¹⁹)_u-NR¹³R¹⁴(ii) a v is 0 or 1; u is 0 or 1; r¹²Is hydrogen or C_1-6An alkyl group; r¹³And R¹⁴Each independently selected from hydrogen and C_{1- 12}Alkyl radical, C_1-6Alkylcarbonyl group, C_1-6Alkylsulfonyl, aryl C_1-6Alkylcarbonyl group, C_3-7Cycloalkyl carbonyl, - (CH)₂)_k-NR¹⁵R¹⁶Selected from hydroxy, hydroxycarbonyl, cyano, C_{1 -6}C substituted by substituents of alkoxycarbonyl or aryl groups_1-12An alkyl group; r¹³And R¹⁴Together with the nitrogen to which they are attached may optionally form morpholinyl, pyrrolidinyl, piperazinyl or be selected from C_1-6Alkyl or aryl radicals C_1-6Piperazinyl substituted with a substituent of alkoxycarbonyl; k is 2; r¹⁵And R¹⁶Each independently selected from hydrogen and C_1-6Alkyl or aryl radicals C_1-6An alkoxycarbonyl group; k is 2; r¹⁵And R¹⁶Each independently selected from hydrogen and C_1-6Alkyl or aryl radicals C_1-6An alkoxycarbonyl group; r¹⁵And R¹⁶Together with the nitrogen to which they are attached may optionally form morpholinyl or piperazinyl or be C_1-6Alkoxycarbonyl-substituted piperazinyl; aryl is phenyl or phenyl substituted by halogen; heteroaryl is pyridyl, indolyl, oxadiazolyl or tetrazolyl; and each pyridyl, indolyl, oxadiazolyl or tetrazolyl group may optionally be substituted by one group selected from C_1-6Alkyl, aryl or aryl C_1-6Alkyl substituents.

A more preferred group of compounds consists of those of formula (I) or any subgroup thereof, wherein m is 0; n is 1; p is 0(ii) a s is 0; t is 0; x is CHR⁸；R⁸Is hydrogen;is-CR⁹C <; each R⁹Is hydrogen; r¹Is hydrogen; r²Is hydrogen or C_1-6An alkoxy group; r²Is hydrogen or C_1-6An alkoxy group; r³Is hydrogen; r⁴And R⁵Each independently is hydrogen, C_1-6Alkyl or C_1-6An alkoxy group; r⁶Is hydrogen; z is a group selected from (a-1), (a-2), (a-3) or (a-4); and R¹⁰Or R¹¹Each independently selected from hydrogen, hydroxy or hydroxy C_1-6An alkyl group.

The most preferable compounds are compound No. 1, compound No. 21, compound No. 4, compound No. 5, compound No. 36, compound No. 69, compound No. 110, compound No. 111, compound No. 112, compound No. 229, and compound No. 37.

The compounds of formula (I), their pharmaceutically acceptable salts and N-oxides, and stereochemically isomeric forms thereof, may be prepared by conventional methods. The starting materials and some intermediates are known compounds and are commercially available or can be prepared according to conventional reaction procedures well known in the art.

Many such methods of preparation are described in more detail below. Other methods of obtaining the final compound of formula (I) are described in the examples.

Compounds of formula (I) may be prepared by reacting an intermediate of formula (II) with an intermediate of formula (III), wherein W is a suitable leaving group, for example halogen, such as fluoro, chloro, bromo or iodo; or a sulfonyloxy group such as methylsulfonyloxy, 4-methylphenylsulfonyloxy, and the like. The reaction may be carried out in a reaction-inert solvent such as an alcohol, e.g., methanol, ethanol, 2-methoxy-ethanol, propanol, butanol, etc.; ethers such as 4, 4-dioxane, 1' -oxydipropane, and the like; ketones such as 4-methyl-2-pentanone; or N, N-dimethylformamide, nitrobenzene, acetonitrile, acetic acid, and the like. The addition of a suitable base, such as an alkali or alkaline earth metal carbonate or bicarbonate, such as triethylamine or sodium carbonate, may be used to absorb the acid released during the reaction. Small amounts of a suitable metal iodide, such as sodium or potassium iodide, may be added to facilitate the reaction. Agitation can increase the reaction rate. The reaction is conveniently carried out at a temperature in the range from room temperature to the reflux temperature of the reaction mixture and, if desired, under elevated pressure.

Wherein X is CH₂Compounds of formula (I) (referred to herein as compounds of formula (I-a)) may be prepared by converting a compound of formula (I) (referred to herein as compounds of formula (I-b)) wherein X is C (═ O), by reacting a compound of formula (I-b) with lithium aluminium hydride in a suitable solvent, such as tetrahydrofuran.

The compounds of formula (I-a) may also be prepared by reacting the appropriate formaldehyde compound of formula (IV) with the intermediate of formula (V) in the presence of an appropriate reagent, for example sodium borohydride, such as sodium tetrahydroborate or a polymer-supported cyanotrihydroborate, in an appropriate solvent, for example an alcohol, such as methanol.

In the same way, compounds of formula (I) wherein t is 1, referred to herein as compounds of formula (I-c), may be prepared by reacting an intermediate of formula (II) with an appropriate formaldehyde compound of formula (VI).

Compounds of formula (I) wherein s is 1, referred to herein as compounds of formula (I-d), may be prepared by reacting an intermediate of formula (VII) with lithium aluminium hydride in a suitable solvent such as tetrahydrofuran.

The compounds of formula (I) and intermediates of formula (III) may also be interconverted by reactions or functional group transformations known in the art. Many such transformations have been described above. Other examples are the hydrolysis of carboxylic esters to the corresponding carboxylic acids or alcohols; hydrolysis of the amide to the corresponding carboxylic acid or amine; nitrile hydrolysis to the corresponding amide; the amino group on the imidazole or phenyl group can be replaced by hydrogen by a diazotization reaction known in the art followed by replacement of the diazo group with hydrogen; alcohols can be converted into esters and ethers; primary amines can be converted to secondary or tertiary amines; the double bond may be hydrogenated to the corresponding single bond; the iodine group on the phenyl group can be converted to an ester group by insertion of carbon monoxide in the presence of a suitable palladium catalyst.

Wherein X is CH₂M is 0, s is 0 and R³The intermediate of formula (II) which is hydrogen (referred to herein as the intermediate of formula (II-a)) may be prepared by reduction of the nitro group to the amine starting from the intermediate of formula (VIII) in the presence of a metal catalyst such as raney nickel and a suitable reducing agent such as hydrogen in a suitable solvent such as methanol or ethanol.

Wherein X is C (═ O), s is 0 and R³Intermediates of formula (II) which are hydrogen (referred to herein as intermediates of formula (II-b)) may be prepared by reacting an intermediate of formula (IX) with an intermediate of formula (X) in the presence of a suitable reagent such as N' - (ethylcarboximidoyl)) -N, N-dimethyl-1, 3-propanediamine, monohydrochloride (EDC) and 1-hydroxy-1H-benzotriazole (HOBT). This reaction can be carried out in the presence of a base such as triethylamine in a suitable solvent such as a mixture of dichloromethane and tetrahydrofuran.

Intermediates of formula (IV) can be prepared by reacting an intermediate of formula (XI) with lithium aluminum hydride in a suitable solvent such as tetrahydrofuran.

Intermediates of formula (VII) can be prepared by reacting an intermediate of formula (XII) with an intermediate of formula (XIII) in the presence of 2-chloro-1-methylpyridinium iodide and triethylamine in a suitable solvent such as acetonitrile.

The intermediate of formula (VIII) can be prepared by reacting an intermediate of formula (XIV) with an intermediate of formula (XV) wherein a is a suitable leaving group,for example halogen, such as fluorine, chlorine, bromine or iodine; or C_1-6Alkoxy, such as methoxy).

Intermediates of formula (XII) may be prepared by converting an intermediate of formula (XVI) in the presence of sodium hydroxide and water in a suitable solvent such as ethanol.

The intermediate of formula (XVI) can be prepared by reacting an intermediate of formula (XVIII), wherein a is as defined above, with an intermediate of formula (XIV) in a suitable solvent such as diisopropylethylamine.

The compounds of formula (I) and some intermediates may have at least one stereogenic center in their structure. Such stereogenic centers may be present in the R or S configuration.

The compounds of formula (I) prepared as described above are generally racemic mixtures of enantiomers which can be separated from each other according to resolution methods well known in the art. The racemic compounds of formula (I) can be converted into the corresponding diastereomeric salt forms by reaction with an appropriate chiral acid. The diastereomeric salt forms are then separated, for example by selective or fractional crystallization, while the enantiomers are liberated therefrom by means of a base. Another method for separating the enantiomeric forms of the compounds of formula (I) involves liquid chromatography using a chiral stationary phase. The pure stereochemically isomeric forms may also be derived from the corresponding pure stereochemically isomeric forms of the appropriate starting materials, provided that the reaction occurs stereospecifically. Preferably, if a particular stereoisomer is desired, the compound will be synthesized by stereospecific methods of preparation. Such a process would advantageously use enantiomerically pure starting materials.

The compounds of formula (I), their pharmaceutically acceptable acid addition salts and stereoisomeric forms have valuable pharmacological properties in the inhibition of the interaction between p53 and MDM 2.

The term "MDM 2" as used herein means the protein obtained as a result of expression of the MDM2 gene. Within the meaning of this term, MDM2 encompasses all proteins encoded by MDM2, variants thereof, selective cleavage proteins thereof, and phosphorylated proteins thereof. In addition, as used herein, the term "MDM 2" includes MDM2 analogs, such as MDMX (also known as MDM4) and MDM2 homologs, and analogs of other animals, such as the human homolog HDm²Or the human analog HDMX.

The term "inhibiting an interaction" or "inhibitor of an interaction" as used herein means avoiding or reducing direct or indirect binding of one or more molecules, peptides, proteins, enzymes or receptors; or to avoid or reduce the normal activity of one or more molecules, peptides, proteins, enzymes or receptors.

The term "inhibitor of the interaction of p53 with MDM 2" or "p 53-MDM2 inhibitor" as used herein is an agent that describes an increase in p53 expression in an assay as described in c.1. Such an increase may be caused by, but is not limited to, one or more of the following mechanisms:

inhibition of the interaction between p53 and MDM2,

direct binding to either of the MDM2 or p53 proteins,

interaction with an upstream or downstream target, such as a kinase, or an enzymatic activity involving ubiquitination or SUMO modification,

isolation or transmission of MDM2 from p53 between different cell compartments,

modulation of proteins binding to MDM2, such as (but not limited to) p73, E2F-1, Rb, p21waf1 or cip1,

down-regulating or interfering with MDM2 expression and/or MDM2 activity, for example by (but not limited to) affecting its cellular localization, post-translational modification, nuclear export or ubiquitin ligase activity,

direct or indirect stabilization of the p53 protein, for example by keeping it in its structural form or by avoiding misfolding,

promoting the expression of p53 or of p53 family members (e.g. p63 and p73),

increasing p53 activity, for example by (but not limited to) enhancing its transcriptional activity and/or

Increase of the expression of genes and proteins of the p 53-signalling pathway, such as, but not limited to, p21waf1, cip1, MIC-1(GDF-15), PIG-3 and ATF-3.

Accordingly, the present invention discloses the use of a compound of formula (I) as a medicament.

In addition, the present invention also relates to the use of a compound for the manufacture of a medicament for the treatment of a disorder mediated via a p53-MDM2 interaction, wherein the compound is a compound of formula (I).

The term "treatment" as used herein encompasses treatment of diseases and/or symptoms in animals, particularly humans, and includes: (i) preventing the disease and/or symptoms from occurring in an individual who may be susceptible to the disease and/or symptoms but has not yet been diagnosed as having it; (ii) inhibiting the disease and/or symptoms, i.e., arresting its development; (iii) relieving the disease and/or symptoms, i.e., causing reversal of the disease and/or symptoms.

The term "disorder mediated via p53-MDM2 interaction" means any undesirable or detrimental symptom that results in or results from inhibition of the interaction between MDM2 protein and p53 or other cellular proteins that induce apoptosis, induce cell death, or regulate the cell cycle.

The present invention also provides a method of treating disorders mediated through the p53-MDM2 interaction by administering an effective amount of a compound of the invention to an individual, such as a mammal (and particularly a human), in need of such treatment.

The compounds of the invention may have an antiproliferative effect in tumour cells, even if such cells do not have functional p 53. More particularly, the compounds of the invention may have an antiproliferative effect in tumours with wild-type p53 and/or in tumours overexpressing MDM 2.

Accordingly, the present invention also provides a method of inhibiting tumor growth by administering an effective amount of a compound of the present invention to a subject, e.g., a mammal (and especially a human) in need of such treatment.

Examples of tumors that can be inhibited are, but are not limited to, lung cancer (e.g., adenocarcinoma and including non-small cell lung cancer), pancreatic cancer (e.g., pancreatic cancer such as exocrine pancreatic cancer), colon cancer (e.g., colorectal cancer such as colon adenocarcinoma and colon adenoma), esophageal cancer, oral squamous cancer, tongue cancer, gastric cancer, nasopharyngeal cancer, hematopoietic tumors of lymphoid lineage (e.g., acute lymphocytic leukemia, B-cell lymphoma, Burkitt's lymphoma), myeloid leukemia (e.g., Acute Myelogenous Leukemia (AML)), thyroid carcinoma, cystoma, myelodysplastic syndrome (MDS), tumors of mesenchymal origin (e.g., myofibrillary and rhabdomyosarcoma), melanoma, teratoma, neuroblastoma, brain tumors, glioma, benign tumors of the skin (e.g., large keratoacanthoma), breast cancer (e.g., advanced breast cancer), Kidney cancer, ovarian cancer, cervical cancer, endometrial cancer, bladder cancer, prostate cancer (including end stage disease), testicular cancer, osteosarcoma, head and neck cancer, and epidermal carcinoma.

The compounds of the invention are also useful in the treatment and prevention of inflammatory conditions.

Accordingly, the present invention also provides a method of treating and preventing inflammatory conditions by administering an effective amount of a compound of the present invention to a subject, such as a mammal (and especially a human) in need of such treatment.

The compounds of the invention are also useful in the treatment of autoimmune diseases and conditions. The term "autoimmune disease" means a disease in which the animal's immune system reacts adversely to self-antigens. The term "autoantigen" means any antigen normally found in an animal. Representative autoimmune diseases include, but are not limited to: hashimoto's thyroiditis, graves ' disease, multiple sclerosis, fatal anemia, addison's disease, insulin-dependent diabetes mellitus, rheumatoid arthritis, systemic lupus erythematosus (SLE or lupus), dermatomyositis, crohn's disease, wagen's granulomatosis, glomerular basement membrane disease, antiphospholipid syndrome, 25 papular dermatitis, allergic encephalomyelitis, glomerulonephritis, membranous glomerulonephritis, goodbud's syndrome, myasthenia-like syndrome (Lambert-Eaton), myasthenia syndrome, myasthenia gravis, pemphigoid, multiple endocrine adenosis, reiter's disease, and stevensan (Stiff-Man) syndrome.

Accordingly, the present invention also provides a method of treating autoimmune diseases and conditions, and diseases associated with conformationally unstable or misfolded proteins, by administering an effective amount of a compound of the invention to a subject, such as a mammal (and particularly a human) in need of such treatment.

The compounds of the present invention are also useful in methods of treating diseases associated with conformationally unstable or misfolded proteins. Examples of diseases associated with conformationally unstable or misfolded proteins include, but are not limited to: cystic Fibrosis (CFTR), marfanstan's syndrome (myofibrillar proteins), amyotrophic lateral sclerosis (superoxide dismutase), scurvy (collagen), maple syrup urine disease (alpha-ketoacid dehydrogenase complex), osteogenesis imperfecta (pre-alpha collagen element), Creutzfeldt-Jakob disease (prion), alzheimer's disease (beta-amyloid fiber), familial amyloidosis (lysozyme), cataracts (crystallin), familial hypercholesterolemia (LDL receptor), alpha-I-antitrypsin deficiency, Tay-Sachs disease (beta-hexosaminidase), retinitis pigmentosa (rhodopsin), and leprosy syndrome (insulin receptor).

Accordingly, the present invention also provides a method of treating a disease associated with conformationally unstable or misfolded proteins by administering an effective amount of a compound of the invention to a subject, such as a mammal (and particularly a human) in need of such treatment.

In view of their useful pharmacological properties, the subject compounds may be formulated into various pharmaceutical forms for administration purposes.

To prepare the pharmaceutical compositions of this invention, an effective amount of the particular compound, in base or acid addition salt form, as the active ingredient is combined in intimate admixture with a pharmaceutically acceptable carrier, which carrier may take a wide variety of forms depending on the form of preparation desired for administration. Such pharmaceutical compositions are desirably in a single dosage form preferably suitable for administration by oral, enteral, transdermal or parenteral injection. For example, in preparing the compositions in oral dosage form, any of the usual pharmaceutical media may be employed, such as water, glycols, oils, alcohols, and the like in the case of oral liquid preparations (e.g., suspensions, syrups, elixirs, and solutions); or in the case of powders, pills, capsules and tablets, solid carriers such as starches, sugars, kaolin, lubricants, binders, disintegrating agents and the like.

Because of their ease of administration, tablets and capsules represent the most advantageous oral dosage unit form in which case solid pharmaceutical carriers are obviously employed. For parenteral compositions, the carrier will typically comprise sterile water (at least in large part) to, for example, aid dissolution, although other ingredients may be included. Injectable solutions, for example, may be prepared in which the carrier comprises saline solution, glucose solution, or a mixture of saline and glucose solution. Injectable suspensions may also be prepared in which case appropriate liquid carriers, suspending agents and the like may be employed. In compositions suitable for transdermal administration, the carrier optionally comprises a penetration enhancer and/or a suitable wetting agent, optionally in combination with a small proportion of suitable additives of any nature which do not cause a significant deleterious effect on the skin. The additives may facilitate administration to the skin and/or may aid in the preparation of the desired composition. Such compositions may be administered in various ways, for example as a transdermal patch, as drops, as a cream. It is particularly advantageous to formulate the above pharmaceutical compositions in dosage unit form for ease of administration and uniformity of dosage. Dosage unit form as used in the specification and claims herein refers to physically discrete units suitable as unitary dosages, each unit containing a predetermined quantity of active ingredient calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier. Examples of such dosage unit forms are tablets (including scored or coated tablets), capsules, pills, powder packets, wafers, injectable solutions or suspensions, teaspoonfuls, tablespoonfuls and the like, and segregated multiples thereof.

It is particularly advantageous to formulate the above pharmaceutical compositions in dosage unit form for ease of administration and uniformity of dosage. Dosage unit form as used in the specification and claims herein refers to physically discrete units suitable as unitary dosages, each unit containing a predetermined quantity of active ingredient calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier. Examples of such dosage unit forms are tablets (including scored or coated tablets), capsules, pills, powder packets, wafers, injectable solutions or suspensions, teaspoonfuls, tablespoonfuls and the like, and segregated multiples thereof.

The compounds of the present invention are administered in an amount sufficient to inhibit the interaction between MDM2 and p53 or other cellular proteins that induce apoptosis, induce cell death, or regulate the cell cycle.

The oncogenic potential of MDM2 is measured not only by its ability to inhibit p53, but also by its ability to modulate other tumor suppressor proteins, such as the retinal glioma protein pRb and the closely related E2F1 transcription factor.

Thus, the compounds of the invention are administered in an amount sufficient to modulate the interaction between MDM2 and the E2F transcription factor.

The effective amount can be readily determined by one skilled in the art from the experimental results presented hereinafter. A therapeutically effective amount is generally considered to be from 0.005mg/kg to 100mg/kg body weight, in particular from 0.005mg/kg to 10mg/kg body weight. It may be appropriate to administer the required dose as one, two, three, four or more sub-doses at appropriate intervals throughout the day. Such sub-doses may be formulated in unit dosage forms, for example containing from 0.5 to 500mg, especially from 10mg to 500mg, of active ingredient per unit dosage form.

As a further aspect of the invention, combinations of a p53-MDM2 inhibitor with another anti-cancer agent are envisaged, in particular for use as a medicament, more in particular for the treatment of cancer or related diseases.

For the treatment of the above-mentioned diseases, the compounds of the invention can advantageously be used in combination with one or more other pharmaceutical agents, more particularly with other anticancer agents. Examples of anticancer agents are:

-platinum coordination compounds, such as cisplatin, carboplatin or oteracil (oxalyplatin);

-paclitaxel compounds, such as paclitaxel or european paclitaxel;

-topoisomerase I inhibitors, such as camptothecin compounds, for example irinotecan or topotecan;

-topoisomerase II inhibitors, such as anti-tumour podophyllotoxin derivatives, for example etoposide or teniposide;

anti-tumour vinca alkaloids, such as vinblastine, vincristine or vinorelbine;

-antineoplastic nucleoside derivatives, such as 5-fluorouracil, gemcitabine or capecitabine;

alkylating agents, such as nitrogen mustard gas or nitrosoureas, such as cyclophosphamide, chlorambucil, carmustine or lomustine;

anti-tumor anthracycline derivatives, such as daunorubicin, doxorubicin, idarubicin, or mitoxantrone;

-HER2 antibodies, such as trastuzumab;

-estrogen receptor antagonists or selective estrogen receptor modulators, such as tamoxifen, toremifene, droloxifene, fischerokee (foslodex) or raloxifene;

aromatase inhibitors, such as exemestane, anastrozole, letrozole (letrozole) and vorozole;

differentiation agents, such as retinoids, vitamin D and Retinoic Acid Metabolism Blockers (RAMBA), such as alcytone (acutane);

-DNA methyltransferase inhibitors, such as azacytidine;

kinase inhibitors, such as flavopiridol (flavoperidol), imatinib mesylate or gefitinib (gefitinib);

-farnesol (farnesyl) transferase inhibitors;

-an HDAC inhibitor;

inhibitors of other ubiquitin-proteasome pathways, such as for example Velcade; or

-Yondelis。

The term "platinum coordination compound" as used herein denotes any tumor cell growth inhibiting platinum coordination compound which provides platinum in ionic form.

The term "paclitaxel compounds" denotes a class of compounds having a paclitaxel ring system and which are related to or derived from extracts of certain species of taxus species;

the term "topoisomerase inhibitor" is used to denote an enzyme capable of altering the DNA topology in eukaryotic cells. Which is critical for important cellular functions and cell proliferation. There are two types of topoisomerase in eukaryotic cells, i.e., type I and type II. Topoisomerase I is a monomeric enzyme of about 100,000 molecular weight. The enzyme binds to DNA and produces a short single strand break, unwinds (or unwinds) the double helix, and then reseals the break before separating from the DNA strand. Topoisomerase II has a similar mechanism of action, which is involved in causing breaks in DNA strands or formation of free radicals.

The term "camptothecin compound" is used to denote a compound related to or derived from the parent camptothecin compound, which is a water-insoluble alkaloid derived from camptotheca acuminata in china and the cute branches of india.

The term "podophyllotoxin compound" is used to denote a compound related to or derived from a parent podophyllotoxin, which is extracted from the datura stramonium plant.

The term "antitumor vinca alkaloids" is used to indicate extracts related to or derived from the vinca plant (day spring).

The term "alkylating agent" encompasses a diverse group of chemicals having a common property that has the ability to donate alkyl groups under physiological conditions to macromolecules such as DNA necessary to sustain life on an organism. For most of the more important agents, such as nitrogen mustard gas and nitrosoureas, the active alkylating moiety is produced in vivo after complex degradation reactions, which are in part enzymatic. The most important pharmacological actions of alkylating agents are those that disrupt the fundamental mechanisms associated with cell proliferation, particularly DNA synthesis and cell division. In rapidly proliferating tissues, the ability of alkylating agents to interfere with DNA function and integrity provides the basis for their therapeutic applications and many of their toxic properties.

The term "antineoplastic anthracycline derivatives" encompasses antibiotics obtained from the mould strep. peuticus var. caesius and derivatives thereof, characterized by having a tetracycline ring structure with an unusual sugar, a hexammine sugar, connected by glycosidic linkages.

The expansion of the human epidermal growth factor receptor 2 protein (HER 2) in primary breast cancer has been shown to be associated with poor clinical prognosis in some patients. Trastuzumab is a highly purified recombinant DNA-derived humanized monoclonal IgG1 kappa antibody that binds with high affinity and specificity to the extracellular region of the HER2 receptor.

Many breast cancers have estrogen receptors and the growth of such tumors can be stimulated by estrogen. The terms "estrogen receptor antagonist" and "selective estrogen receptor modulator" are used to denote competitive inhibitors of estradiol binding to the Estrogen Receptor (ER). Selective estrogen receptor modulators, when bound to the ER, cause a change in the three-dimensional shape of the receptor, regulating its binding to Estrogen Responsive Elements (ERE) on DNA.

In climacteric women, the main source of circulating estrogen is derived from the conversion of adrenal and ovarian male hormones (androstenedione and testosterone) to estrogen (estrone and estradiol) by aromatase in peripheral tissues. Estrogen loss via aromatase inhibition or inactivation is an effective and alternative treatment for certain climacteric patients with hormone-dependent breast cancer.

The term "antiestrogen" is intended to include not only estrogen receptor antagonists and selective estrogen receptor modulators, but also aromatase inhibitors as described above.

The term "differentiation agent" encompasses compounds that inhibit cell proliferation and cause differentiation in a variety of ways. Vitamin D and retinoids are known to play a major role in regulating the growth and differentiation of various normal and malignant cell types. Retinoic acid metabolism blockers (RAMBA's) increase the concentration of endogenous retinoic acid by inhibiting cytochrome P450-mediated catabolism of retinoic acid.

DNA methylation changes are the most common abnormalities in human neoplasia. Hypermethylation within the promoter of a selected gene is often associated with inactivation of the gene of interest. The term "DNA methyltransferase inhibitor" is used to indicate a compound that acts via pharmacological inhibition of DNA methyltransferase and regeneration of tumor suppressor gene expression.

The term "kinase inhibitor" encompasses potent inhibitors of kinases associated with cell cycle progression and programmed cell death (apoptosis).

The term "farnesol transferase inhibitor" is used to denote a compound designed to prevent farnesylation of Ras and other intracellular proteins. It has been demonstrated to have an effect on malignant cell proliferation and survival.

The term "histone deacetylase inhibitor" or "inhibitor of histone deacetylase" is used to identify compounds that are capable of interacting with histone deacetylase and inhibiting their activity, in particular their enzymatic activity. Inhibiting histone deacetylase enzymatic activity means reducing the ability of histone deacetylase to remove acyl groups from histone.

The term "inhibitors of other ubiquitin-proteasome pathways" is used to identify compounds that inhibit the targeted disruption of cellular proteins (including cell cycle regulatory proteins) in the protein plastid.

As mentioned above, the compounds of the invention also have therapeutic applications in sensitizing tumor cells for chemotherapy and radiotherapy.

Thus, the compounds of the present invention may be used as "radiosensitizers" and/or "chemosensitizers" or may be used in combination with another "radiosensitizer" and/or "chemosensitizer".

The term "radiosensitizer," as used herein, is defined as a molecule, preferably a low molecular weight molecule, administered to an animal in a therapeutically effective amount to increase the sensitivity of cells to ionizing radiation and/or to promote the treatment of diseases treatable with ionizing radiation.

The term "chemosensitizer", as used herein, is defined as a molecule, preferably a low molecular weight molecule, administered to an animal in a therapeutically effective amount to increase the sensitivity of cells to chemotherapy and/or to promote the treatment of diseases treatable by chemotherapy.

Several mechanisms of the mode of action of radiosensitizers have been disclosed in the literature, including: hypoxic cell radiosensitizers (e.g., 2-nitroimidazole compounds and benzotriazine dioxide compounds) mimic oxygen or otherwise act like bioreductive agents in the absence of oxygen; the non-hypoxic cell radiosensitizer (e.g., halogenated pyrimidine) can be an analog of a DNA base, and is preferably introduced into the DNA of cancer cells, thereby promoting radiation-induced fragmentation of the DNA molecule and/or preventing normal DNA repair mechanisms; and various other possible mechanisms that have been hypothesized to be the action of radiosensitizers in the treatment of disease.

Many current cancer treatment protocols use radiation sensitive agents in combination with x-ray radiation. Examples of x-ray activated radiosensitizers include, but are not limited to, the following compounds: metronidazole, misonidazole, desmethonidazole, pimonidazole, etanidazole, nimozole, mitomycin C, RSU 1069, SR 4233, EO9, RB 6145, niacinamide, 5-bromodeoxyuridine (BUdR), 5-iododeoxyuridine (IUdR), bromodeoxycytidine, fluorodeoxyuridine (FudR), hydroxyurea, cisplatin, and therapeutically effective analogs and derivatives thereof.

Photodynamic therapy (PDT) of cancer employs visible light as a radiation activator of a sensitizer. Examples of photodynamic radiation-sensitive agents include, but are not limited to, the following compounds: hematoporphyrin derivatives, Photofrin (Photofrin), benzoporphyrin derivatives, stannorphyrin, umberbyl-a (phenobibide-a), bacteriochlorophyll-a, naphthalocyanine, phthalocyanine, zinc phthalocyanine and therapeutically effective analogs and derivatives thereof.

The radiosensitizer can be administered in combination with a therapeutically effective amount of one or more other compounds, including but not limited to: a compound that promotes binding of the radiosensitizer to the target cell; compounds that control the flow of therapeutics, nutrients, and/or oxygen to target cells; chemotherapeutic agents that act on tumors with or without additional radiation; or other therapeutically effective compounds for the treatment of cancer or other diseases.

The chemosensitive agent may be administered in combination with a therapeutically effective amount of one or more other compounds, including but not limited to: a compound that promotes the binding of chemosensitizers into target cells; compounds that control the flow of therapeutics, nutrients, and/or oxygen to target cells; chemotherapeutic agents that act on tumors or other therapeutically effective compounds for the treatment of cancer or other diseases.

In view of their useful pharmacological properties, the components of the combination according to the invention, i.e. the further agent and the p53-MDM inhibitor, may be formulated in various pharmaceutical forms for administration purposes. The components may be formulated separately in each pharmaceutical composition or in a single pharmaceutical composition comprising both components.

The invention therefore also relates to a pharmaceutical composition comprising a further drug and a p53-MDM inhibitor together with one or more pharmaceutical carriers.

The invention further relates to the use of a combination according to the invention for the manufacture of a pharmaceutical composition for inhibiting the growth of tumor cells.

The invention further relates to a product comprising as a first active ingredient a p53-MDM2 inhibitor according to the invention and as a second active ingredient an anti-cancer agent, as a combined preparation for simultaneous, separate or sequential use in the treatment of a patient suffering from cancer.

The other agent and the p53-MDM2 inhibitor may be administered simultaneously (e.g., separately or in a single composition) or sequentially in any order. In the latter case, the two compounds will be administered in an amount and manner sufficient to ensure that a beneficial or synergistic effect is achieved over a period of time. It will be appreciated that the preferred method and sequence of administration of the components of the combination, as well as the respective dosages and therapies, will depend upon the particular other agent being administered, as well as the p53-MDM2 inhibitor, its route of administration, the particular tumor being treated, and the particular host being treated. The optimal method and sequence of administration and dosage and therapy can be readily determined by those skilled in the art using conventional methods and with reference to the information set forth herein.

Platinum coordination compounds are advantageousIs administered in an amount of 1 to 500mg per square meter of body surface area (mg/m)²) E.g. 50 to 400mg/m²In particular for cisplatin, the dose per course of treatment is about 75mg/m²And a dosage per course of treatment of about 300mg/m for carboplatin²。

Advantageous doses of paclitaxel compound are 50 to 400mg per square meter of body surface area (mg/m)²) E.g. 75 to 250mg/m²In particular, the dosage per course of treatment for paclitaxel is about 175 to 250mg/m²And for European taxol, the dosage per treatment course is about 75 to 150mg/m²。

Camptothecin compounds are advantageously administered in a dose of 0.1 to 400mg per square meter of body surface area (mg/m)²) E.g. 1 to 300mg/m²And in particular for irinotecan, at a dose of about 100 to 350mg/m per course of treatment²And for topotecan, the dosage per course of treatment is about 1 to 2mg/m²。

The antitumor podophyllotoxin derivative is advantageously administered in a dose of 30 to 300mg (mg/m) per square meter of body surface area²) E.g. 50 to 250mg/m²In particular for etoposide, from about 35 to 100mg/m per course of treatment²And a dosage per course of treatment of about 50 to 250mg/m for teniposide²。

The antitumor alkaloid is advantageously administered in a dose of 2 to 30mg per square meter of body surface area (mg/m)²) In particular for vinblastine in a dosage of about 3 to 12mg/m per course of treatment²The dose per course of treatment for vincristine is about 1 to 2mg/m²And a dose per treatment course of vinorelbine of about 10 to 30mg/m²。

The antitumor nucleoside derivatives are advantageously administered in a dose of 200 to 2500mg per square meter of body surface area (mg/m)²) For example 700 to 1500mg/m²In particular, for 5-FU, the dosage per treatment course is about 200 to 500mg/m²The dose per course of treatment for gemcitabine is about 800 to 1200mg/m²And a dosage per course of treatment of about 1000 to 2500mg/m for capecitabine²。

Alkylating agents such as nitrogen mustard gas or nitrosourea are advantageously administered in a dose of 100 to 500mg per square meter of body surface area (mg/m)²) E.g. 120 to 200mg/m²In particular, for cyclophosphamide, the dose per course is about 100 to 500mg/m²The dosage per course of treatment for chlorambucil is about 0.1 to 0.2mg/kg, and for carmustine is about 150 to 200mg/m²And a dose per course of treatment of about 100 to 150mg/m for lomustine²。

Advantageous doses of antitumor anthracycline derivatives are from 10 to 75mg per square meter of body surface area (mg/m)²) E.g. 15 to 60mg/m²In particular, for doxorubicin, the dosage per course of treatment is about 40 to 75mg/m²For daunorubicin, the dosage per course of treatment is about 25 to 45mg/m²And a dosage per course of treatment of about 10 to 15mg/m for idarubicin²。

Trastuzumab is advantageously administered in a dose of 1 to 5mg per square meter of body surface area (mg/m)²) In particular 2 to 4mg/m per course of treatment²。

The antiestrogens are advantageously administered in a dosage of about 1 to 100mg per day, depending on the particular agent and the condition to be treated. Tamoxifen is advantageously orally administered twice a day at a dose of 5 to 50mg, preferably 10 to 20mg, for a sufficient time of treatment to achieve and maintain the therapeutic effect. Toremifene is advantageously administered orally at a dose of about 60mg once a day. Anastrozole is advantageously administered orally at a dose of about 1mg once a day. Droloxifene is advantageously administered orally at a dose of about 20-100mg once a day. Raloxifene is advantageously administered orally at a dose of about 60mg once a day. Exemestane is advantageously administered orally at a dose of about 25mg once a day.

These doses may be administered, for example, once, twice or more per course of treatment, which may be repeated, for example, every 7, 14, 21 or 28 days.

The compounds of formula (I), their pharmaceutically acceptable acid addition salts and stereoisomeric forms thereof, may have valuable diagnostic properties in that they may be used to detect or identify p53-MDM2 interactions in biological samples, including the detection or measurement of the formation of complexes between marker compounds and/or p53 and/or MDM2 and/or other molecules, peptides, proteins, enzymes or receptors.

The detection or identification method may use a compound labeled with a labeling agent such as a radioisotope, an enzyme, a fluorescent substance, a luminescent substance, or the like. Examples of radioactive isotopes include¹²⁵I、¹³¹I、³H and¹⁴C. enzymes are typically made detectable by binding to an appropriate substrate, which in turn catalyzes a detectable reaction. Examples thereof include, for example, β -galactosidase, β -glucosidase, alkaline phosphatase, peroxidase and malate dehydrogenase, preferably horseradish peroxidase. Luminescent substances include, for example, luminol derivatives, luciferin, aequorin, and luciferase. A biological sample may be defined as a body tissue or a body fluid. Examples of body fluids are cerebrospinal fluid, blood, plasma, serum, urine, sputum, saliva, etc.

The following examples illustrate the invention.

Experimental part

Hereinafter, "DMF" is defined as N, N-dimethylformamide, "DCM" is defined as dichloromethane, "DIPE" is defined as diisopropyl ether, "EtOAc" is defined as ethyl acetate, "EtOH" is defined as ethanol, "EDC" is defined as N' - (ethylcarboximidoyl) -N, N-dimethyl-1, 3-propanediamine, monohydrochloride, "MeOH" is defined as methanol, "THF" is defined as tetrahydrofuran, and "HOBT" is defined as 1-hydroxybenzotriazole.

A. Preparation of intermediate compounds

Example A1

a) Preparation of intermediate 1

A mixture of 1-fluoro-4-nitro-benzene (0.0142mol), 1H-indole-3-ethylamine (0.0129mol) and N-ethyl-N- (1-methylethyl) -2-propylamine (0.032mol) was stirred at 210 ℃ for 18 hours, then allowed to return to room temperature and decanted. The residue was taken up in acetonitrile/water. The precipitate was filtered, washed with ether and dried, yielding 2.3g (64%) of intermediate 1.

b) Preparation of intermediate 2

A mixture of intermediate 1(0.0078mol) and raney nickel (2.2g) in EtOH (50ml) was hydrogenated at room temperature under a pressure of 3 bar for 3 hours and then filtered through Celite (Celite). The celite was washed with DCM/MeOH. The filtrate was evaporated. The residue was taken up in DCM and dried (MgSO₄) Filtration and evaporation of the solvent gave 1.88g (95%) of intermediate 2.

Example A2

a) Preparation of intermediate 3

A mixture of 2-ethoxy-1-methoxy-1-nitro-benzene (0.009mol), 1H-indole-3-ethylamine (0.009mol) and N-ethyl-N- (1-methylethyl) -2-propylamine (0.0228mol) was stirred at 210 ℃ for 24 hours, then allowed to come to room temperature, taken up in DCM/MeOH and dried. The residue was taken up in DCM (small amount) and purified by column chromatography over silica gel (35-70 μm) (eluent: cyclohexane/DCM 30/70). The pure fractions were collected and the solvent was evaporated, yielding 0.6g (20%) of intermediate 3.

b) Preparation of intermediate 4

Intermediate 3(0.002mol) and H were combined at room temperature₂A mixture of Raney nickel (0.6g) in MeOH (100ml) was hydrogenated at a pressure of 3 bar for 1 hour 30 minutes and then filtered through celite. The celite was washed with DCM/MeOH. The filtrate was evaporated. The residue was taken up in DCM/MeOH (aq) and dried (MgSO)₄) Filtration and evaporation of the solvent gave 0.45g (83%) of intermediate 4.

Example A3

a) Preparation of intermediate 5

A mixture of N-3-pyridyl-acetamide (0.038mol), 1-fluoro-4-nitro-benzene (0.05mol), copper (I) chloride (0.0038mol) and potassium carbonate (0.076mol) in xylene was stirred under reflux for 18 hours and then allowed to return to room temperature. Water was added. The mixture was filtered through celite. The celite was washed with DCM. The filtrate was evaporated. The residue was purified by column chromatography over silica gel (35-70 μm) (eluent: DCM/MeOH/NH)₄OH 97/3/0.1). The pure fractions were collected and the solvent was evaporated, yielding 6.4g (65%) of intermediate 5.

b) Preparation of intermediate 6

Sodium hydroxide (concentrated) (10ml) was added to a mixture of intermediate 5(0.025mol) in EtOH (80 ml). The mixture was stirred and refluxed for 2 hours, then allowed to return to room temperature. Water was added. The mixture was stirred for 15 minutes and then filtered. The residue was purified by column chromatography over silica gel (70-200 μm) (eluent: DCM/MeOH 100/0-95/5). The pure fractions were collected and the solvent was evaporated, yielding 1.5g (28%) of intermediate 6.

c) Preparation of intermediate 7

A mixture of intermediate 6(0.007mol) and Raney nickel (1.5g) in MeOH (30ml) and THF (10ml) was hydrogenated at room temperature under a pressure of 3 bar for 1 hour, then filtered through celite. The celite was washed with DCM/MeOH. The filtrate was evaporated. The residue was taken up in DCM. The organic layer was separated and dried (MgSO)₄) Filtration and evaporation of the solvent gave 1.2g (92%) of intermediate 7.

Example A4

Preparation of intermediate 8

1H-indole-3-propionic acid (0.0264mol), then 1-hydroxybenzotriazole (0.0344mol), then N' - (ethylcarboximidoyl) -N, N-dimethyl-1, 3-propanediamine, monohydrochloride (═ EDCI) (0.0344mol) was added to a mixture of 1, 4-phenylenediamine (0.137mol) in THF (200ml) and DCM (200ml) under a stream of nitrogen. The mixture was stirred at room temperature for 24 hours, poured into water and extracted with DCM. The organic layer was separated and dried (MgSO)₄) Filtered and the solvent evaporated. The residue was purified by column chromatography over silica gel (20-45 μm) (eluent: DCM/MeOH/NH)₄OH 97/3/0.5). The pure fractions were collected and the solvent was evaporated, yielding 1.75g (24%) of intermediate 8.

Example A5

a) Preparation of intermediate 9

A mixture of 1-fluoro-4-nitro-benzene (0.0025mol), 1-methyl-1H-indol-3-ethylamine (0.0023mol) and N-ethyl-N- (1-methylethyl) -2-propylamine (0.0057mol) was stirred at 200 ℃ for 2 hours and then allowed to return to room temperature. Water and DCM were added. The organic layer was separated and dried (MgSO)₄) Filtered and the solvent evaporated. The residue (0.8g) was purified by column chromatography over silica gel (35-70 μm) (eluent: DCM 100). The pure fractions were collected and the solvent was evaporated. The residue (0.45g) was taken up in DIPE. The precipitate was filtered and dried, yielding 0.33g (66%) of intermediate 9.

b) Preparation of intermediate 10

A mixture of intermediate 9(0.0011mol) and raney nickel (0.4g) in MeOH (20ml) was hydrogenated at room temperature under a pressure of 3 bar for 1 hour, then filtered through celite. The celite was washed with DCM/MeOH. The filtrate was evaporated. The residue was taken up in DCM. The organic layer was separated and dried (MgSO)₄) Filtration and evaporation of the solvent gave 0.305g (97%) of intermediate 10.

Example A6

a) Preparation of intermediate 11

A mixture of 4-fluoro-benzonitrile (0.071mol), 1H-indole-3-ethylamine (0.071mol) and N-ethyl-N- (1-methylethyl) -2-propylamine (0.1775mol) was stirred at 210 ℃ for 16H, then allowed to return to room temperature and taken up in DCM/MeOH. The organic layer was washed with 3N HCl and dried (MgSO)₄) Filtered and the solvent evaporated. The residue was collected in ether/acetonitrile. The precipitate was filtered and dried, yielding 8.07g (43%) of intermediate 11, mp 144 ℃.

b) Preparation of intermediate 12

A mixture of intermediate 11(0.0115mol) and sodium hydroxide (0.17mol) in EtOH (50ml) and water (50ml) was stirred and refluxed for 18 hours, then allowed to return to room temperature. The solvent was evaporated. The residue was taken up in 3N sodium hydroxide. The aqueous layer was washed with DCM and acidified until pH 5 was obtained. The precipitate was filtered and dried, yielding 1.06g (35%) of intermediate 12, mp 225 ℃.

c) Preparation of intermediate 13

A mixture of intermediate 12(0.0037mol), 4-pyridylamine (0.0037mol), 2-chloro-1-methyl-pyridinium iodide (0.0113mol) and triethylamine (0.015mol) in acetonitrile (100ml) was stirred andrefluxed for 90 minutes and then allowed to return to room temperature. The solvent was evaporated. The residue was taken up in DCM/MeOH. The organic layer was washed with potassium carbonate 10% and dried (MgSO)₄) Filtered and the solvent evaporated until dry. The residue was purified by flash column chromatography over silica gel (35-70 μm) (eluent: DCM/MeOH/NH)₄OH 95/5/0.1). Two fractions were collected and the solvent was evaporated, yielding 0.06g F1 and 0.08g F2. F1 was crystallized from diethyl ether/acetonitrile. The precipitate was filtered and dried to yield a first batch of 0.032g (24%) of intermediate 13. The F2 and mother liquor layers were combined and crystallized from ether/acetonitrile. The precipitate was filtered and dried to yield a second crop of 0.105g (10%) of intermediate 13, m.p. 200 ℃.

Example A7

Preparation of intermediate 14

Following a procedure analogous to method 6 (see example a13), starting from 1- (4-chloro-2-pyridinyl) -ethanone (227mg, 0.0015mol) and adding triethylamine (0.22 ml). After work-up, the residue was purified by column chromatography over silica gel (40-63 μm) (eluent: EtOAc). The pure fractions were collected and the solvent was evaporated, yielding 256mg (52%) of intermediate 14 as a pale yellow oil.

Example A8

Preparation of intermediate 15

Following a procedure analogous to method 4 (see example A22/22), starting from benzyl 1-piperazinecarboxylate (1.3ml, 0.0069mol) and 4-chloro-2-pyridinemethanol (500mg, 0.0034 mol). After work-up, the residue was purified by column chromatography over silica gel (40-63 μm) (eluent: DCM/MeOH 95/5). The pure fractions were collected and the solvent was evaporated, yielding 840mg (70%) of intermediate 15 as an orange oil.

Example A9

Preparation of intermediate 16

Following a procedure analogous to method 4 (see example A22/22), starting from 4-amino-butan-1-ol (310mg, 0.0021mol) and 4-chloro-2-pyridinemethanol (300mg, 0.0021 mol). After work-up, the residue was purified by column chromatography over silica gel (40-63 μm) (eluent: DCM/MeOH 85/15). The pure fractions were collected and the solvent was evaporated, yielding 55mg (17%) of intermediate 16 as a yellow oil.

Example A10

Preparation of intermediate 17

Following a procedure analogous to method 5 (see example a22/34), starting from 4- (2-amino-ethyl) -piperazine-1-carboxylic acid tert-butyl ester (579mg, 0.0025mol) and 4-chloro-2-pyridinecarboxaldehyde (325mg, 0.0023 mol). After work-up, the residue was purified by column chromatography over silica gel (40-63 μm) (eluent: DCM/MeOH 90/10). The pure fractions were collected and the solvent was evaporated, yielding 425mg (53%) of intermediate 17 as a yellow oil.

Example A11

Preparation of intermediate 18

Following a procedure analogous to method 5 (see example A22/34), starting from 3-amino-propionic acid methyl ester hydrochloride (351mg, 0.0025mol) and 4-chloro-2-pyridinecarboxaldehyde (325mg, 0.0023mol) and triethylamine (0.35ml, 0.0025mol) was added. After work-up, the residue was purified by column chromatography over silica gel (40-63 μm) (eluent: DCM/MeOH 95/5). The pure fractions were collected and the solvent was evaporated, yielding 160mg (30%) of intermediate 18 as a yellow oil.

Example A12

Preparation of intermediate 19

Similar procedure as in method 3 (see example A22/20) was followed, starting from bromo-methyl acetate (0.26ml, 0.0021mol) and 4-chloro-2-pyridinemethanol (300mg, 0.0021 mol). After work-up, the residue was purified by column chromatography over silica gel (40-63 μm) (eluent: AcOEt/cyclohexane 60/40). The pure fractions were collected and the solvent was evaporated, yielding 170mg (38%) of intermediate 19 as a yellow oil.

Example A13

Preparation of intermediate 20

Method 6

4-amino-1-butanol (0.13ml, 0.0014mol) was added to 1- (4-chloro-2-pyridinyl) -ethanone (200mg, 0.0013mol), p-toluenesulfonic acid (123mg, 0.00065mol) and 3 at room temperatureMolecular sieves in a mixture in MeOH (4 ml). The mixture was stirred at room temperature for 6 h, cooled to 0 ℃ and sodium borohydride (98mg, 0.0026mol) was added slowly. The mixture was stirred at room temperature for 18 hours. The molecular sieve was filtered off, the mixture was poured into water and the solvent was evaporated. The aqueous layer was basified with a saturated solution of sodium bicarbonate and extracted 3 times with DCM. The organic layer was separated, washed with brine, and dried (MgSO)₄) Filtered and the solvent evaporated. The residue was purified by column chromatography over silica gel (40-63 μm) (eluent: DCM/MeOH 95/5). The pure fractions were collected and the solvent was evaporated, yielding 269mg (91%) of intermediate 20 as a yellow oil.

Example A14

Preparation of intermediate 21

A mixture of α - (phenylmethyl) -1H-indole-3-acetic acid (94mg, 0.00035mol) and 1, 1-carbonyldiimidazole (59mg, 0.00036mol, added in portions) in DCM (1ml) was stirred at room temperature under argon for 3 hours. N, O-dimethylhydroxylamine hydrochloride (36mg, 0.00037mol) was added and the mixture was stirred at room temperature for a further 3 hours, cooled to 0 ℃ and poured into water. The pH was adjusted to 10 with sodium hydroxide in 4N and the aqueous layer was extracted with EtOAc. The organic layer was separated, washed with 3N hydrochloric acid solution and dried (MgSO)₄) Filtered and the solvent evaporated. The residue was purified by column chromatography over silica gel (40-63 μm) (eluent: EtOAc/cyclohexane 50/50). Collecting the pure fractions and dissolving the solventEvaporation gave 52mg (47%) of intermediate 21.

Example A15

a) Preparation of intermediate 22

4-dimethylaminopyridine (261mg, 0.0021mol) and di-tert-butyl dicarbonate (di-tert-butyldicarbonate) (14.0g, 0.064mol) were added to a solution of intermediate 1(3.0g, 0.011mol) in DCM (130 ml). The mixture was stirred at room temperature for 5 hours. The reaction was quenched by addition of water and extracted 2 times with DCM. The organic layer was washed successively with a saturated solution of sodium bicarbonate and with brine, dried (MgSO)₄) Filtered and the solvent evaporated. The residue was purified by column chromatography over silica gel (40-63 μm) (eluent: EtOAc/cyclohexane 10/90 to 20/80). The pure fractions were collected and the solvent was evaporated, yielding 4.65g (90%) of intermediate 22 as a yellow oil.

b) Preparation of intermediate 23

Raney nickel (3g) was added to a solution of intermediate 22(4.7g, 0.0097mol) in ethanol (15ml) and THF (15 ml). The reaction mixture was stirred under 1 atmosphere of hydrogen for 16 hours. To complete the reaction, Raney nickel (1g) was added and the mixture was stirred under 1 atmosphere of hydrogen for an additional 4 hours. The mixture was filtered through a pad of celite and the solvent was evaporated. The residue was purified by column chromatography over silica gel (40-63 μm) (eluent: EtOAc/cyclohexane 10/90 to 20/80). The pure fractions were collected and the solvent was evaporated, yielding 4.0g (92%) of intermediate 23 as a yellow foam.

Example A16

a) Preparation of intermediate 24

A solution of bromine (0.0104mol) followed by sodium nitrite (0.0362mol) in water (3ml) was added dropwise at-10 ℃ to a mixture of 6, 7-dihydro-5H-1-pyridin-4-amine (0.0112mol) in aqueous hydrogen bromide (48%) (5 ml). The mixture was returned to 20 ℃. Ice is added. The mixture was basified with concentrated sodium hydroxide and extracted with EtOAc. The organic layer was separated and dried (MgSO)₄) Filtration and evaporation of the solvent gave 2g (90%) of intermediate 24.

b) Preparation of intermediate 25

M-chloroperbenzoic acid (0.012mol) was added to a mixture of intermediate 24(0.01mol) in DCM (15 ml). The mixture was stirred at room temperature for 12 hours. 3N sodium hydroxide and water were added. The mixture was extracted 3 times with DCM. The organic layer was washed with water and dried (MgSO)₄) Filtration and evaporation of the solvent gave 1.85g (86%) of intermediate 25.

c) Preparation of intermediate 26

A mixture of intermediate 25(0.0086mol) in acetic anhydride (18ml) was stirred at 100 ℃ for 30 minutes, then cooled to room temperature and evaporated. The residue was taken up in NaHCO₃And EtOAc and filtered through celite. The celite was washed with EtOAc. The organic layer was separated and dried (MgSO)₄) Filtration and evaporation of the solvent gave 1.63g (73%) of intermediate 26.

d) Preparation of intermediate 27

A mixture of intermediate 26(0.0074mol) in MeOH (10ml) and 3N sodium hydroxide (80ml) was stirred at room temperature for 30 min, then at 80 ℃ for 10 min, and then allowed to return to room temperature. MeOH was evaporated. The mixture was extracted 2 times with DCM and then washed with saturated NaCl. The organic layer was separated and dried (MgSO)₄) Filtration and evaporation of the solvent gave 1.02g (64%) of intermediate 27.

Example A17

a) Preparation of intermediate 28

A solution of bromine (1.3ml) followed by sodium nitrite (3.3g) in water (4ml) was added dropwise at-10 ℃ to a solution of 5, 6, 7, 8-tetrahydro-4-quinolinamine (0.0135mol) in aqueous hydrogen bromide (48%) (6.7 ml). The mixture was returned to 20 ℃. Poured out on ice, basified with concentrated sodium hydroxide and extracted with EtOAc. The organic layer was separated and dried (MgSO)₄) Filtration and evaporation of the solvent gave 2.2g (77%) of intermediate 28.

b) Preparation of intermediate 29

M-chloroperbenzoic acid (0.0125mol) was added to a mixture of intermediate 28(0.0104mol) in DCM (20 ml). The mixture was stirred at room temperature for 12 hours. 3N sodium hydroxide and ice were added. The mixture was extracted 2 times with DCM. The organic layer was dried (MgSO₄) Filtration and evaporation of the solvent gave 3g (100%) of intermediate 29.

c) Preparation of intermediate 30

A mixture of intermediate 29(0.0086mol) in acetic anhydride (22ml) was stirred at 100 ℃ for 30 minutes, then cooled to room temperature and evaporated. The residue was taken up in saturated NaHCO₃And EtOAc. The mixture was stirred for 30 minutes. The organic layer was separated and dried (MgSO)₄) Filtration and evaporation of the solvent gave 3.4g (100%) of intermediate 30.

d) Preparation of intermediate 31

A mixture of intermediate 30(0.0104mol) in MeOH (18ml) and 3N sodium hydroxide (150ml) was stirred at room temperature for 30 min, then at 80 ℃ for 10 min. MeOH was evaporated. The mixture was extracted 2 times with DCM. The organic layer was washed with saturated NaCl and dried (MgSO)₄) Filtration and evaporation of the solvent gave 1.84g (77%) of intermediate 31.

Example A18

a) Preparation of intermediate 32

A mixture of 2-ethoxy-4-nitroanisole (0.0107mol), 6-methoxytryptamine (0.0107mol) and diisopropylethylamine (0.0268mol) was stirred at 210 ℃ for 5 hours, then poured onto ice and extracted with DCM. The organic layer was separated and dried (MgSO)₄) Filtered and the solvent evaporated. The residue was purified by column chromatography over silica gel (15-40 μm) (eluent: DCM 100%). The pure fractions were collected and the solvent was evaporated, yielding 0.85g (22%) of intermediate 32.

b) Preparation of intermediate 33

A mixture of intermediate 32(0.0023mol) and raney nickel (0.85g) in MeOH (42ml) and THF (42ml) was hydrogenated at 3 bar pressure for 2 hours at room temperature and then filtered through celite. The filtrate was evaporated, yielding 0.74g (95%) of intermediate 33.

Example A19

a) Preparation of intermediate 34

Oxalyl chloride (0.012mol) was added dropwise to a solution of 5-cyanoindole (0.007mol) in diethyl ether (21ml) at 0 ℃. The mixture was stirred at 0 ℃ for 5 hours and then at room temperature overnight. The precipitate was filtered, washed with ether and dried, yielding 1.454g (73%) of intermediate 34.

b) Of intermediate 35Preparation of

A solution of intermediate 34(0.0027mol) in DCM (12ml) was added dropwise to a solution of N-pyridin-4-yl-benzene-1, 4-diamine (0.022mol) and N, N-diisopropylethylamine (0.0034mol) in DCM (4ml) at 5 ℃. The mixture was stirred and refluxed over the weekend, then cooled to room temperature. The precipitate was filtered and dried. The residue was crystallized from iPrOH. The precipitate was filtered and dried to yield 0.756g of crude product. This fraction was purified by column chromatography over kromasil (5 μm) (eluent: DCM/MeOH/NH)₄OH97/3/0.3 to 87/13/1.3). The pure fractions were collected and the solvent was evaporated. The residue was purified by column chromatography over silica gel (15-40 μm) (eluent: DCM/MeOH/NH)₄OH 90/10/0.1 to 87/13/0.1). The pure fractions were collected and the solvent was evaporated, yielding 0.098g (20%) of intermediate 35, melting point > 264 ℃.

Example A20

a) Preparation of intermediate 36

A mixture of 1H-benzimidazole-1-ethylamine (0.011mol), 1-fluoro-4-nitro-benzene (0.011mol) and diisopropylethylamine (0.034mol) was stirred at 210 ℃ for 30 minutes. Diisopropylethylamine was evaporated. The precipitate was dissolved in DCM/MeOH. The organic layer was washed with 10% potassium carbonate and dried (MgSO)₄) Filtered and the solvent evaporated. The residue (3.2g) was purified by column chromatography over silica gel (15-40 μm) (eluent: DCM/MeOH/NH)₄OH 98/2/0.5). The pure fractions were collected and the solvent was evaporated. The residue was crystallized from acetonitrile (2.1g, 77%). The precipitate was filtered and dried to yield 1.3g (47%) of intermediate 36, meltPoint 144 ℃.

b) Preparation of intermediate 37

A mixture of intermediate 36(0.006mol) and raney nickel (2g) in MeOH (20ml) was hydrogenated at room temperature at a pressure of 3 bar and then filtered through celite. The celite was washed with DCM/MeOH. The filtrate was evaporated, yielding 1.7g (100%) of intermediate 37.

Example A21

a) Preparation of intermediate 38

A mixture of DL-tryptophan methyl ester (0.0078mol), 1-fluoro-4-nitro-benzene (0.0078mol) and diisopropylethylamine (0.0353mol) was stirred at 210 ℃ for 4 h, then taken up in DCM/MeOH and 3N HCl added. The mixture was stirred for 15 minutes. With saturated NaHCO₃The organic layer was washed and dried (MgSO)₄) Filtered and the solvent evaporated. The residue was purified by column chromatography over silica gel (35-70 μm) (eluent: DCM 100% followed by DCM/MeOH 99/1). The pure fractions were collected and the solvent was evaporated, yielding 0.75g (28%) of intermediate 38.

b) Preparation of intermediate 39

A mixture of intermediate 38(0.0022mol) and raney nickel (0.75g) in MeOH (100ml) was hydrogenated at 3 bar pressure for 1 hour at room temperature and then filtered through celite. The filtrate was evaporated, yielding 0.65g (96%) of intermediate 39.

c) Preparation of intermediate 40

A mixture of intermediate 39(0.139mol) and 4-bromopyridine hydrochloride (0.139mol) in acetic acid (450ml) was stirred at 120 ℃ for 3 hours, poured out on ice, basified with concentrated sodium hydroxide and extracted with DCM/MeOH (small amount). The organic layer was separated and dried (MgSO)₄) Filtered and the solvent evaporated. The residue (62.7g) was purified by column chromatography over kromasil (20-45 μm) (eluent: DCM/MeOH/NH)₄OH 93/7/0.5). The pure fractions were collected and the solvent was evaporated, yielding 22g (41%) of intermediate 40.

d) Preparation of intermediate 41

Lithium hydroxide monohydrate (0.112mol) was added portionwise to a solution of intermediate 40(0.056mol) in MeOH (86ml) and water (34.4ml) at 0 ℃ under a stream of nitrogen. The mixture was stirred at room temperature overnight, then evaporated until dry, yielding: 22g (quantitative yield) of intermediate 41.

Example A22

1) Preparation of intermediate 42

A2.5N solution of butyllithium in hexane (3.4ml, 0.0081mol) was added to a solution of diisopropylamine (0.85ml, 0.0088mol) in THF (6ml) at-78 deg.C under argon. The mixture was stirred at-78 ℃ for 30 minutes. 4-chloro-3-methylpyridine hydrochloride (630mg, 0.0038mol) was added in portions and the mixture was stirred at-78 ℃ for 1 hour. Diethyl carbonate (1.0ml, 0.0096mol) was added dropwise and the mixture was stirred at-78 ℃ for a further 1 hour, then warmed to room temperature and stirred for 2.5 hours. The reaction was quenched by slow addition of water and extracted 2 times with EtOAc. The organic layer was separated, washed with brine, and dried (MgSO)₄) Filtered and the solvent evaporated. The residue was purified by column chromatography over silica gel (40-63 μm) (eluent: EtOAc/MeOH 100/0, then 90/10). The pure fractions were collected and the solvent was evaporated, yielding 74mg (9%) of intermediate 42.

2) Preparation of intermediate 43

Triethyl phosphonoacetate (0.075ml, 0.00038mol) was added dropwise to a mixture of sodium hydride (10.6mg, 0.00044mol) in THF (5ml) at room temperature under argon. The mixture was stirred at room temperature for 20 minutes, then a solution of 4-chloro-3-pyridinecarboxaldehyde (50mg, 0.00035mol) in THF (3ml) was added dropwise. The mixture was stirred at room temperature for 16 h, then poured into water and extracted 2 times with EtOAc. The organic layer was separated, washed with brine, and dried (MgSO)₄) Filtration and evaporation of the solvent gave 77mg (88%) of intermediate 43.

3) Preparation of intermediate 44

At-78 ℃ inA2.5N solution of butyllithium in hexane (0.80ml, 0.0020mol) was added to a solution of diisopropylamine (0.28ml, 0.0020mol) in THF (2ml) under argon. The mixture was stirred at-78 ℃ for 10 minutes, then a solution of 4-chloropyridine (219mg, 0.0019mol) in THF (1ml) was added dropwise. The mixture was stirred at-78 ℃ for 1.25 h, then propionaldehyde (0.14ml, 0.0019mol) was added dropwise. The mixture was stirred at-78 ℃ for 30 min, finally at room temperature for 4 h, poured into water and extracted with EtOAc. The organic layer was separated, washed with brine, and dried (MgSO)₄) Filtered and the solvent evaporated. The residue was purified by column chromatography over silica gel (40-63 μm) (eluent: EtOAc/cyclohexane 90/10). The pure fractions were collected and the solvent was evaporated, yielding 147mg (44%) of intermediate 44.

4) Preparation of intermediate 45

Method 2

A3M solution of ethyl magnesium bromide in ether (1.1ml, 0.0032mol) was added to a solution of methyl 4-chloro-2-picolinate (200mg, 0.0012mol) in THF (4ml) at-30 ℃ under argon. The mixture was stirred at 75 ℃ for 2 hours 30 minutes, cooled to 0 ℃ and quenched with water. The resulting mixture was made basic with a saturated solution of sodium bicarbonate and extracted 2 times with EtOAc. The organic layer was washed with brine and dried (MgSO)₄) Filtered and the solvent evaporated. The residue was purified by column chromatography over silica gel (40-63 μm) (eluent: EtOAc/cyclohexane 10/90). The pure fractions were collected and the solvent was evaporated, yielding 54mg (23%) of intermediate 45 as a brown oil.

5) Preparation of intermediate 46

Methanesulfonyl chloride (98. mu.l, 0.0013mol) was added dropwise to a solution of 4-chloro-2-pyridinemethylamine (150mg, 0.0011mol) and triethylamine (177. mu.l, 0.0013mol) in DCM (4ml) at 0 ℃ under argon. The mixture was stirred at room temperature for 30 minutes. The reaction was quenched with a saturated solution of sodium bicarbonate and extracted 2 times with DCM. The organic phase was dried (MgSO)₄) Filtered and the solvent evaporated. The residue was purified by column chromatography over silica gel (40-63 μm) (eluent: EtOAc). The pure fractions were collected and the solvent was evaporated, yielding 82mg (35%) of intermediate 46 as an orange oil.

6) Preparation of intermediate 47

Method 7

Cyclopropanecarbonyl chloride (115. mu.l, 0.0013mol) was added dropwise to a solution of 4-chloro-2-pyridinemethylamine (150mg, 0.0011mol) and triethylamine (177. mu.l, 0.0013mol) in DCM (4ml) at 0 ℃ under argon. The mixture was stirred at room temperature for 15 minutes. The reaction was quenched with a saturated solution of sodium bicarbonate and extracted 2 times with DCM. The organic phase was dried (MgSO)₄) Filtered and the solvent evaporated. The residue was purified by column chromatography over silica gel (40-63 μm) (eluent: EtOAc). The pure fractions were collected and the solvent was evaporated, yielding 85mg (38%) of intermediate 47 as a white solid.

7) Preparation of intermediate 48

Following a procedure analogous to that for method 7 (see example A22/6), starting from 4-chloro-2-pyridinemethanamine (150mg, 0.0011mol) and hydrocinnamoyl chloride (187. mu.l, 0.0013 mol). After work-up, the residue was purified by column chromatography over silica gel (40-63 μm) (eluent: EtOAc). The pure fractions were collected and the solvent was evaporated, yielding 191mg (66%) of intermediate 48 as a yellow solid.

8) Preparation of intermediate 49

Following a procedure analogous to that of method 7 (see example A22/6), starting from 4-chloro-2-pyridinemethylamine (200mg, 0.0014mol) and propionyl chloride (146. mu.l, 0.0017 mol). After work-up, the residue was purified by column chromatography over silica gel (40-63 μm) (eluent: EtOAc). The pure fractions were collected and the solvent was evaporated, yielding 126mg (45%) of intermediate 49 as colorless oil.

9) Preparation of intermediate 50

Following a procedure analogous to that of method 7 (see example A22/6), starting from 4-chloro-2-pyridinemethylamine (150mg, 0.0011mol) and phenylacetyl chloride (168. mu.l, 0.0013 mol). After work-up, the residue was purified by column chromatography over silica gel (40-63 μm) (eluent: EtOAc). The pure fractions were collected and the solvent was evaporated, yielding 124mg (45%) of intermediate 50 as a white solid.

10) Preparation of intermediates 51 and 52

And

intermediate 51 intermediate 52

Method 1

1.4M solution of cyclopropylmagnesium bromide in toluene/THF (75/25) was added dropwise to a solution of 4-chloro-2-pyridinecarboxaldehyde (377mg, 0.0027mol) in THF (4ml) at 0 ℃ under argon. The reaction mixture was stirred at 0 ℃ for 1 hour, the dry ice bath was removed and the mixture was stirred at room temperature for 1 hour. The reaction mixture was quenched by addition of water and extracted 2 times with EtOAc. The organic layer was washed with brine and dried (MgSO₄) Filtered and the solvent evaporated. The residue was purified by column chromatography over silica gel (40-63 μm) (eluent: cyclohexane/EtOAc 80/20). The pure fractions were collected and the solvent was evaporated, yielding 193mg (39%) of intermediate 51 and 29mg of intermediate 52.

11) Preparation of intermediate 53

Following a procedure analogous to method 1 (see example A22/10), starting from 4-chloro-2-pyridinecarboxaldehyde (300mg, 0.0021mol) and a 2.0M solution of isopropylmagnesium chloride in THF (2.12ml, 0.0042 mol). After work-up, the residue was purified by column chromatography over silica gel (40-63 μm) (eluent: cyclohexane/EtOAc 80/20). The pure fractions were collected and the solvent was evaporated, yielding 165mg (42%) of intermediate 53 as a brown oil.

12) Preparation of intermediate 54

Following a procedure analogous to method 6 (see example a13), starting from 1- (4-chloro-2-pyridine) -ethanone (298mg, 0.0019 mol). After work-up, the residue was purified by column chromatography over silica gel (40-63 μm) (eluent: cyclohexane/EtOAc 90/10). The pure fractions were collected and the solvent was evaporated, yielding 293mg (62%) of intermediate 54 as a yellow oil.

13) Preparation of intermediate 55

Following a procedure analogous to method 6 (see example a13), starting from 1- (4-chloro-2-pyridine) -ethanone (100mg, 0.00064 mol). After work-up, the residue was purified by column chromatography over silica gel (40-63 μm) (eluent: DCM/MeOH 85/15). The pure fractions were collected and the solvent was evaporated, yielding 50mg (45%) of intermediate 55 as a yellow oil.

14) Preparation of intermediate 56

Following a procedure analogous to method 6 (see example a13), starting from 1- (4-chloro-2-pyridine) -ethanone (100mg, 0.00064 mol). After work-up, the residue was purified by column chromatography over silica gel (40-63 μm) (eluent: DCM/MeOH 90/10). The pure fractions were collected and the solvent was evaporated, yielding 30mg (25%) of intermediate 56 as a yellow oil.

15) Preparation of intermediate 57

Following a procedure analogous to method 6 (see example a13), starting from 1- (4-chloro-2-pyridine) -ethanone (157mg, 0.0010 mol). After work-up, the residue was purified by column chromatography over silica gel (40-63 μm) (eluent: DCM/MeOH 95/5). The pure fractions were collected and the solvent was evaporated, yielding 134mg (49%) of intermediate 57 as a yellow oil.

16) Preparation of intermediate 58

Following a procedure analogous to method 6 (see example a13), starting from 1- (4-chloro-2-pyridinyl) -ethanone (200mg, 0.0013 mol). After work-up, the residue was purified by column chromatography over silica gel (40-63 μm) (eluent: DCM/MeOH 95/5). The pure fractions were collected and the solvent was evaporated, yielding 281mg (66%) of intermediate 58 as a yellow oil.

17) Preparation of intermediate 59

Following a procedure analogous to method 6 (see example a13), starting from 1- (4-chloro-2-pyridinyl) -ethanone (200mg, 0.0013mol) and adding triethylamine (0.2 ml). After work-up, the residue was purified by column chromatography over silica gel (40-63 μm) (eluent: EtOAc). The pure fractions were collected and the solvent was evaporated, yielding 80mg (25%) of intermediate 59 as a yellow oil.

18) Preparation of intermediate 60

Following a procedure analogous to method 6 (see example a13), starting from 1- (4-chloro-2-pyridinyl) -ethanone (200mg, 0.0013 mol). After work-up, the residue was purified by column chromatography over silica gel (40-63 μm) (eluent: DCM/MeOH 95/5). The pure fractions were collected and the solvent was evaporated, yielding 176mg (68%) of intermediate 60 as a pale yellow oil.

19) Preparation of intermediate 61

Following a procedure analogous to method 6 (see example a13), starting from 1- (4-chloro-2-pyridinyl) -ethanone (200mg, 0.0013 mol). After work-up, the residue was purified by column chromatography over silica gel (40-63 μm) (eluent: EtOAc). The pure fractions were collected and the solvent was evaporated, yielding 274mg (77%) of intermediate 61 as a yellow oil.

20) Preparation of intermediate 62

Method 3

4-chloro- α -methyl-2-pyridinemethanol (200mg, 0.0013mol) in THF (3ml) was added dropwise to a mixture of sodium hydride (60% by weight in mineral oil) (56mg, 0.0014mol) in THF (1ml) at 0 ℃ under argon. The mixture was heated to 70 ℃ and stirred for 3 hours, then cooled to 0 ℃ and iodoethane (0.102ml, 0.0013mol) was added dropwise. The mixture was heated to 70 ℃ for 2 hours, cooled to 0 ℃, poured out on ice water and extracted 2 times with DCMAnd extracted 1 time with EtOAc. The organic layer was separated and dried (MgSO)₄) Filtered and the solvent evaporated. The residue was purified by column chromatography over silica gel (40-63 μm) (eluent: cyclohexane/EtOAc 90/10). The pure fractions were collected and the solvent was evaporated, yielding 106mg (45%) of intermediate 62 as a brown oil.

21) Preparation of intermediate 63

Following a procedure analogous to method 3 (see example A22/20), starting from 4-chloro-. alpha. -methyl-2-pyridinemethanol (200mg, 0.0013 mol). After work-up, the residue was purified by column chromatography over silica gel (40-63 μm) (eluent: cyclohexane/EtOAc 90/10). The pure fractions were collected and the solvent was evaporated, yielding 85mg (39%) of intermediate 63 as a pale yellow oil.

22) Preparation of intermediate 64

Method 4

4-chloro-2-pyridinemethanol (400mg, 0.0028mol) was dissolved in chloroform (24 ml). Sulfuryl chloride (0.40ml, 0.0056mol) and DMF (2 drops) were added. The mixture was stirred at 80 ℃ for 4 hours. The solvent was removed by evaporation. The residue was taken up back into MeOH (18ml) and ethanolamine (1.38ml, 0.014mol) was added. The mixture was stirred at 80 ℃ for 4 hours. The solvent was removed by evaporation. The residue was poured into water and extracted with EtOAc. The organic layer was separated, washed with a saturated solution of sodium bicarbonate and dried (MgSO)₄) Filtered and the solvent evaporated. The residue was purified by column chromatography over silica gel (40-63 μm) (eluent: DCM/MeOH 85/15). Collecting pure fractions and purifyingThe solvent was evaporated, yielding 310mg (60%) of intermediate 64 as an orange oil.

23) Preparation of intermediate 65

Following a procedure analogous to that of method 4 (see example A22/22), starting from 2-morpholin-4-yl-ethylamine (0.45ml, 0.0034mol) and 4-chloro-2-pyridinemethanol (200mg, 0.0014 mol). After work-up, the residue was purified by column chromatography over silica gel (40-63 μm) (eluent: DCM/MeOH 95/5). The pure fractions were collected and the solvent was evaporated, yielding 39mg (23%) of intermediate 65 as a yellow oil.

24) Preparation of intermediate 66

Following a procedure analogous to method 4 (see example A22/22), starting from a 33% solution of methylamine in EtOH (10ml) and 4-chloro-2-pyridinemethanol (300mg, 0.0021 mol). After work-up, the residue was purified by column chromatography over silica gel (40-63 μm) (eluent: DCM/NH)₄OH 85/15/1). The pure fractions were collected and the solvent was evaporated, yielding 130mg (40%) of intermediate 66 as an orange oil.

25) Preparation of intermediate 67

Following a procedure analogous to method 4 (see example A22/22), starting from a 2.0M solution of ethylamine in THF (3.5ml, 0.0069mol) and 4-chloro-2-pyridinemethanol (200mg, 0.014 mol). After work-up, the residue was purified by column chromatography over silica gel (40-63 μm) (eluent: DCM/MeOH 85/15). The pure fractions were collected and the solvent was evaporated, yielding 45mg (19%) of intermediate 67 as an orange oil.

26) Preparation of intermediate 68

Following a procedure analogous to method 4 (see example A22/22), starting from ethyl 3-amino-propionate (2.45g, 0.020mol) and 4-chloro-2-pyridinemethanol (600mg, 0.0042 mol). After work-up, the residue was purified by column chromatography over silica gel (40-63 μm) (eluent: DCM/MeOH 85/15). The pure fractions were collected and the solvent was evaporated, yielding 730mg (71%) of intermediate 68 as an orange liquid.

27) Preparation of intermediate 69

Intermediate 68(350mg, 0.0015mol) was dissolved in MeOH (5ml) and cooled to 0 ℃. Sodium borohydride (300mg, 0.0078mol) was added slowly. The mixture was stirred at 80 ℃ for 9 hours. The reaction was quenched with water and the solvent was evaporated. The residue was extracted with EtOAc. The organic layer was separated, washed with a saturated solution of sodium bicarbonate and dried (MgSO)₄) Filtered and the solvent evaporated. The residue was purified by column chromatography over silica gel (40-63 μm) (eluent: DCM/MeOH 85/15). The pure fractions were collected and the solvent was evaporated, yielding 70mg (23%) of intermediate 69 as a colorless oil.

28) Preparation of intermediate 70

Similar procedure as in method 4 (see example A22/22) was followed, starting from amino-acetonitrile (1.2g, 0.013mol) and 4-chloro-2-pyridinemethanol (500mg, 0.0034 mol). After work-up, the residue was purified by column chromatography over silica gel (40-63 μm) (eluent: DCM/MeOH 95/5). The pure fractions were collected and the solvent was evaporated, yielding 160mg (25%) of intermediate 70 as an orange liquid.

29) Preparation of intermediate 71

Similar procedure as in method 4 (see example A22/22) was followed, starting from N-methylpiperazine (1.16ml, 0.010mol) and 4-chloro-2-pyridinemethanol (300mg, 0.0021 mol). After work-up, the residue was purified by column chromatography over silica gel (40-63 μm) (eluent: DCM/MeOH 95/5). The pure fractions were collected and the solvent was evaporated, yielding 325mg (69%) of intermediate 71 as a yellow oil.

30) Preparation of intermediate 72

Similar procedure as in method 4 (see example A22/22) was followed, starting from diethylamine (1.45ml, 0.014mol) and 4-chloro-2-pyridinemethanol (300mg, 0.0021 mol). After work-up, the residue was purified by column chromatography over silica gel (40-63 μm) (eluent: DCM/MeOH 95/5). The pure fractions were collected and the solvent was evaporated, yielding 300mg (43%) of intermediate 72 as a yellow liquid.

31) Preparation of intermediate 73

Similar procedure as in method 4 (see example A22/22) was followed, starting from 3-aminopropionitrile (1.02ml, 0.014mol) and 4-chloro-2-pyridinemethanol (500mg, 0.0034 mol). After work-up, the residue was purified by column chromatography over silica gel (40-63 μm) (eluent: DCM/MeOH 95/5). The pure fractions were collected and the solvent was evaporated, yielding 180mg (27%) of intermediate 73 as a yellow oil.

32) Preparation of intermediate 74

Similar procedure as in method 4 (see example A22/22) was followed, starting from phenethylamine (0.52ml, 0.0042mol) and 4-chloro-2-pyridinemethanol (300mg, 0.0021 mol). After work-up, the residue was purified by column chromatography over silica gel (40-63 μm) (eluent: EtOAc). The pure fractions were collected and the solvent was evaporated, yielding 110mg (22%) of intermediate 74 as a colorless oil.

33) Preparation of intermediate 75

Similar procedure as in method 4 (see example A22/22) was followed, starting from 3-phenyl-propylamine (470mg, 0.0035mol) and 4-chloro-2-pyridinemethanol (300mg, 0.0021 mol). After work-up, the residue was purified by column chromatography over silica gel (40-63 μm) (eluent: DCM/MeOH 85/15). The pure fractions were collected and the solvent was evaporated, yielding 90mg (17%) of intermediate 75 as an orange oil.

34) Preparation of intermediate 76

Method 5

4-chloro-2-pyridinecarboxaldehyde (200mg, 0.0014mol), N- (3-aminopropyl) morpholine (224mg, 0.0015mol), p-toluenesulfonic acid (134mg, 0.00070mol) andthe molecular sieve was stirred at room temperature under argon for 7 hours. The molecular sieve was filtered off, the reaction mixture was cooled to 0 ℃ and sodium borohydride (107mg, 0.0028mol) was added slowly. The mixture was stirred at room temperature for 17 hours, poured into water and extracted with DCM. The organic layer was separated, washed with a saturated solution of sodium bicarbonate and dried (MgSO)₄) Filtered and the solvent evaporated. The residue was purified by column chromatography over silica gel (40-63 μm) (eluent: DCM/MeOH/NH)₃85/15/3). The pure fractions were collected and the solvent was evaporated, yielding 230mg (60%) of intermediate 76 as a yellow oil.

35) Preparation of intermediate 77

Similar procedure as in method 4 (see example A22/22) was followed, starting from benzylamine (0.46ml, 0.0042mol) and 4-chloro-2-pyridinemethanol (300mg, 0.0021 mol). After work-up, the residue was purified by column chromatography over silica gel (40-63 μm) (eluent: EtOAc/cyclohexane 50/50). The pure fractions were collected and the solvent was evaporated, yielding 240mg (50%) of intermediate 77 as colorless oil.

36) Preparation of intermediate 78

Similar procedure as in method 3 (see example A22/20) was followed, starting from bromoethyl methyl ether (0.13ml, 0.0014mol) and 4-chloro-2-pyridinemethanol (200mg, 0.0014 mol). After work-up, the residue was purified by column chromatography over silica gel (40-63 μm) (eluent: AcOEt/cyclohexane 30/70). The pure fractions were collected and the solvent was evaporated, yielding 67mg (24%) of intermediate 78 as a yellow oil.

37) Preparation of intermediate 79

Similar procedure as in method 4 (see example A22/22) was followed, starting from 4-phenyl-butylamine (0.55ml, 0.0035mol) and 4-chloro-2-pyridinemethanol (250mg, 0.0017 mol). After work-up, the residue was purified by column chromatography over silica gel (40-63 μm) (eluent: DCM/MeOH 95/5). The pure fractions were collected and the solvent was evaporated, yielding 260mg (55%) of intermediate 79 as a yellow oil.

38) Preparation of intermediate 80

Following a procedure analogous to that of method 3 (see example A22/20), starting from (3-bromo-propyl) -benzene (0.27ml, 0.0018mol) and 4-chloro-2-pyridinemethanol (200mg, 0.0014 mol). After work-up, the residue was purified by column chromatography over silica gel (40-63 μm) (eluent: AcOEt/cyclohexane 10/90). The pure fractions were collected and the solvent was evaporated, yielding 57mg (16%) of intermediate 80 as a colorless oil.

39) Preparation of intermediate 81

Following a procedure analogous to that of method 3(cA22/20), starting from 1-bromo-2-ethoxy-ethane (589mg, 0.0052mol) and 4-chloro-2-pyridinemethanol (500mg, 0.0034 mol). After work-up, the residue was purified by column chromatography over silica gel (40-63 μm) (eluent: EtOAc/cyclohexane 10/90). The pure fractions were collected and the solvent was evaporated, yielding 270mg (36%) of intermediate 81 as a colorless oil.

40) Preparation of intermediate 82

Similar procedure as in method 1 (see example A22/10) was followed, starting from 4-chloro-2-pyridinecarboxaldehyde (500mg, 0.0035 mol). After work-up, the residue was purified by column chromatography over silica gel (40-63 μm) (eluent: EtOAc/cyclohexane 50/50). The pure fractions were collected and the solvent was evaporated, yielding 136mg (22%) of intermediate 82 as a yellow oil.

Example A23

a) Preparation of intermediate 83

A37% hydrochloric acid solution (14.3ml) was added to a solution of 2-ethoxy-4-nitro-aniline (10.5g, 0.0577mol) in acetic acid (210ml), and the mixture was stirred at room temperature for 30 minutes. A solution of sodium nitrite (4.4g, 0.0635mol) in water (15ml) was then added dropwise and the mixture was stirred at 0 ℃ for 30 minutes. A cold solution of potassium iodide (19.2g, 0.1157mol) and iodine (7.3g, 0.0288mol) in water (70ml) was added dropwise at 0 ℃. The mixture was stirred at 0 ℃ for 30 minutes and at room temperature for 16 hours. The precipitate formed was filtered off, washed with water and then dissolved in DCM. The organic solution was washed with a saturated solution of sodium bicarbonate and dried (MgSO)₄) Filtration and evaporation of the solvent gave 13.7g (81%) of intermediate 83 as a yellow solid.

b) Preparation of intermediate 84

A mixture of intermediate 83(700mg, 0.0024mol), 6-methoxytryptamine (505mg, 0.0026mol), dichloro [1, 1 '-bis (diphenylphosphino) ferrocene ] palladium (II) dichloromethane adduct (78mg, 0.00011mol), 1' -bis (diphenylphosphino) ferrocene (177mg, 0.00032mol) and sodium tert-butoxide (255mg, 0.0026mol) in THF (95ml) was heated at 100 ℃ for 3 hours and at 120 ℃ for 1.5 hours. After filtration through a pad of celite, the solvent was evaporated and the residue was purified by column chromatography over silica gel (40-63 μm) (eluent: DCM). The pure fractions were collected and the solvent was evaporated, yielding 464mg (55%) of intermediate 84 as a yellow solid.

c) Preparation of intermediate 85

A mixture of intermediate 84 (see example A23/b) (773mg, 0.0022mol) and Raney nickel (50% slurry in water) in ethanol (8.5ml) and THF (6.8ml) was stirred at room temperature under 1 atmosphere of hydrogen for 24 hours. After filtration through a pad of celite, the solvent was evaporated, yielding 697mg (98%) of intermediate 85 as a purple foam.

Example A24

Preparation of intermediates 86 and 87

Intermediate 86

Andintermediate 87

A mixture of 3, 4, 5-trichloro-pyridazine (200mg, 0.0011mol), intermediate 2 (see example A1/b) (273mg, 0.0011mol) and diisopropylamine (0.38ml, 0.0011mol) was stirred in 2-propanol (4.0ml) at 80 ℃ for 1 h. The solvent was evaporated and the crude mixture was taken back up to EtOAc. The organic layer was washed with a saturated solution of sodium bicarbonate and brine, dried (MgSO)₄) Filtered and the solvent evaporated. The residue was purified by column chromatography over silica gel (40-63 μm) (eluent: EtOAc/cyclohexane 50/50). The pure fractions were collected and the solvent was evaporated, yielding 179mg (41%) of intermediate 86 and intermediate 87 as 1/1 mixtures of the two pyridazine compounds.

Example A25

a) Preparation of intermediate 88

4-Oxiranyl-1- (phenylmethyl) -piperidine (0.069mol) was stirred in MeOH (300ml) and NaOCH₃(0.069mol) and refluxing for 6 hours. The solvent was evaporated, then the residue was taken up in water and extracted with DCM. The organic layer was separated, dried, filtered and the solvent was evaporated. The residue was purified by column chromatography over silica gel (eluent: DCM/MeOH98/2, 90/10, 85/15). The product fractions were collected and the solvent was evaporated, yielding 5.0g (29%) of intermediate 88.

b) Preparation of intermediate 89

A mixture of intermediate 88 (see example A25/a) (0.02mol) in MeOH (100ml) was hydrogenated over Pd/C10% (1g) as catalyst. In absorption of H₂After (1 eq.) the catalyst was filtered off and the filtrate was evaporated, yielding 3.18g (100%) of intermediate 89.

Example A26

a) Preparation of intermediate 90

Similar procedure as in method 5 (see example A22/34) was followed, starting from benzyl N- (2-aminoethyl) carbamate hydrochloride (475ml, 0.0020mol) and 4-chloro-2-pyridinecarbaldehyde (265mg, 0.0019mol) and adding triethylamine (0.29ml, 0.0021 mol). After work-up, the residue was purified by column chromatography over silica gel (40-63 μm) (eluent: DCM/MeOH 95/5). The pure fractions were collected and the solvent was evaporated, yielding 150mg (25%) of intermediate 90 as a colorless oil.

B. Finalization ofPreparation of the Compounds

Example B1

Preparation of Compound 1

A mixture of 4-chloro-quinoline (0.0009mol) and intermediate 2(0.001mol) in 2-propanol (5ml) was stirred and refluxed for 6 hours, then allowed to come to room temperature. The solvent was evaporated. The residue was basified with 10% potassium carbonate and extracted with DCM. The organic layer was separated and dried (MgSO)₄) Filtered and the solvent evaporated. The residue (0.38g) was purified by column chromatography over silica gel (10 μm) (eluent: DCM/MeOH/NH)₄OH 97/3/0.5). The pure fractions were collected and the solvent was evaporated. 2-propanol and HCl/2-propanol were added. The mixture was stirred for 30 minutes and allowed to return to room temperature. The precipitate was filtered and dried with diethyl ether to yield 0.09g (23%) of compound 1, m.p. 170 ℃.

Example B2

Preparation of Compound 2

A mixture of 4-bromo-pyridine hydrochloride (0.0044mol) and intermediate 4(0.0044mol) in acetic acid (13ml) was stirred at 110 ℃ for 45 min, then cooled to room temperature, poured onto ice water, basified with potassium carbonate and extracted with DCM. The organic layer was separated and dried (MgSO)₄) Filtered and the solvent evaporated. The residue (1.4g) was purified by column chromatography over silica gel (15-40 μm) (eluent: DCM/MeOH/NH)₄OH 93/7/0.5). The pure fractions were collected and the solvent was evaporated. The residue (0.38g) was dissolved in 2-propanol/diethyl ether and transferredTo become the hydrochloride salt. The precipitate was filtered and dried, yielding 0.385g (20%) of compound 2, m.p. 150 ℃.

Example B3

Preparation of Compound 3

The mixture of 4-chloro-2 (1H) -quinolinone (0.0011mol) and intermediate 2(0.0016mol) was stirred at 130 ℃ for 5 hours, then stirred at 160 ℃ overnight and allowed to come to room temperature. The residue was purified by column chromatography over silica gel (35-70 μm) (eluent: DCM/MeOH/NH)₄OH 95/5/0.1). The pure fractions were collected and the solvent was evaporated. The residue (0.12g) was taken up in acetonitrile. The precipitate was filtered and dried, yielding 0.045g (10%) of compound 3, mp 238 ℃.

Example B4

Preparation of Compound 4

Stirring 4-chloro-6, 7-dihydro-5H-cyclopentane [ b ] at 100 deg.C]A mixture of pyridine (0.0006mol) and intermediate 2(0.0006mol) in acetic acid (2ml) was left for 30 minutes and then allowed to come to room temperature. Water was added, followed by sodium hydroxide (3N) and the resulting mixture was extracted with DCM. The organic layer was separated and dried (MgSO)₄) Filtered and the solvent evaporated. The residue obtained (0.233g) was purified by column chromatography over silica gel (10 μm) (eluent: DCM/MeOH/NH)₄OH 97/3/0.3). The pure fractions were collected and the solvent was evaporated, yielding 0.025g (11%) of compound 4.

Example B5

Preparation of Compound 5

A mixture of 4-chloro-5, 6, 7, 8-tetrahydro-quinoline (0.0009mol) and intermediate 2(0.0009mol) in DMF (3ml) was stirred at 100 ℃ for 3 h and then allowed to come to room temperature. The mixture was poured into ice water and sodium hydroxide (3N) and then extracted with DCM. The organic layer was separated and dried (MgSO)₄) Filtered and the solvent evaporated. The residue obtained (0.49g) was purified by column chromatography over silica gel (5 μm) (eluent: DCM/MeOH/NH)₄OH 99/1/0.05 to 80/20/0.5). The pure fractions were collected and the solvent was evaporated, yielding 0.054g (16%) of compound 5.

Example B6

Preparation of Compound 6

Lithium aluminium hydride (0.0032mol) was added portionwise to a mixture of N-methoxy-N-methyl-1H-indole-3-acetamide (0.0032mol) in THF (5ml) at 0 ℃ under a stream of N2. The mixture was stirred for 1 hour. Potassium hydrogen sulfate (5%) was added. The mixture was extracted with diethyl ether. The organic layer was separated and dried (MgSO)₄) Filtered and the solvent evaporated. This mixture must be used immediately. Intermediate 7(0.0016mol) in MeOH (5ml), in a polymeric vehicle (Amberlite IRA-300 BH)₃CN form-Capacity BH₃CN ═ 2.5/4.5mmol/g resin) cyanoborohydride (0.0022mol) and acetic acid (several drops) were added to the obtained mixture. The mixture was stirred for 12 hours. The precipitate was filtered and dried. The residue was purified by column chromatography over silica gel (15-40 microns)Spectral purification (eluent: DCM/MeOH/NH)₄OH 97/3/0.3). The pure fractions were collected and the solvent was evaporated. The residue (0.14g) was taken up in HCl/2-propanol. The precipitate was filtered and dried, yielding 0.125g (29%) of compound 6, mp 160 ℃.

Example B7

Preparation of Compound 7

A mixture of N-4-pyridyl-1, 4-phenylenediamine (0.0016mol) and 6-methoxy-1H-indole-3-carbaldehyde (0.0016mol) in MeOH (20ml) was stirred and refluxed overnight. Sodium tetrahydroborate (0.0016mol) was added. The mixture was stirred at room temperature for 4 hours, poured onto ice and extracted with DCM. The organic layer was separated and dried (MgSO)₄) Filtered and the solvent evaporated. The residue was purified by column chromatography over kromasil (10 μm) (eluent: DCM/MeOH/NH)₄OH 92/8/0.5 followed by toluene/2-propanol/NH₄OH 85/15/1). The pure fractions were collected and the solvent was evaporated. The residue was crystallized from acetonitrile. The precipitate was filtered and dried, yielding 0.131g (23%) of compound 7, m.p. 145 ℃.

Example B8

Preparation of Compound 8

A mixture of 4-bromo-pyridine hydrochloride (0.0069mol) and intermediate 8(0.008mol) in acetic acid (7ml) was stirred at 120 ℃ for 1 hour and then allowed to come to room temperature. Water was added. The mixture was basified with potassium carbonate and extracted 2 times with DCM/MeOH (95/5). The organic layer was separated and dried (MgSO)₄) Filtered and the solvent evaporated. The residue was purified by flash column chromatography over silica gel (35-70 μm) (eluent: DCM/MeOH/NH)₄OH 92/8/0.5). The pure fractions were collected and the solvent was evaporated, yielding 1.6g (65%) of compound 8, m.p. 208 ℃.

Example B9

Preparation of Compound 9

4-Pyridinecarboxaldehyde (0.0005mol) was then added to a polymeric support (Amberlite IRA-300 BH)₃CN form-Capacity BH₃CN- ═ 2.5/4.5mmol/g resin) cyanoborohydride (0.0004mol), then acetic acid (3 drops) was added to a mixture of intermediate 2(0.0004mol) in MeOH (10 ml). The mixture was stirred at room temperature for 3 hours. The precipitate was filtered and washed with MeOH. The filtrate was evaporated. The residue (0.17g) was dissolved in MeOH (20ml) and was a mixture of the title compound 9 and the corresponding unreduced intermediate imine. Sodium tetrahydroborate (0.02g) was added in portions. The mixture was stirred for 30 minutes. Water was added. MeOH was partially evaporated. The mixture was extracted with EtOAc. The organic layer was washed with water and dried (MgSO)₄) Filtered and the solvent evaporated. The residue (0.05g) was purified by column chromatography over silica gel (10 μm) (eluent: DCM/MeOH/NH)₄OH 98/2/0.4). The pure fractions were collected and the solvent was evaporated. The residue (0.034g) was dissolved in 2-propanone and converted to the oxalate salt. The precipitate was filtered and dried, yielding 0.036g (16%) of compound 9, m.p. 132 ℃.

Example B10

Preparation of Compound 10

A mixture of 4-bromo-pyridine hydrochloride (0.001mol) and intermediate 10(0.0005mol) in acetic acid (2ml) was stirred at 120 ℃ for 1 hour and then allowed to return to room temperature. Ice was added followed by 3N sodium hydroxide. The mixture was extracted 2 times with DCM. The organic layer was separated and dried (MgSO)₄) Filtered and the solvent evaporated. The residue was purified by column chromatography over silica gel (10 μm) (eluent: DCM/MeOH/NH)₄OH 96/4/0.5). The pure fractions were collected and the solvent was evaporated. The residue (0.064g, 29%) was dissolved in 2-propanol/diethyl ether and converted to the hydrochloride salt. The precipitate was filtered and dried, yielding 0.082g (29%) of compound 10, m.p. > 250 ℃.

Example B11

Preparation of Compound 11

Lithium aluminium hydride (0.0145mol) was added to a mixture of intermediate 13(0.0036mol) in THF (100 ml). The mixture was stirred and refluxed for 3 hours, then allowed to return to room temperature. EtOAc was added. A minimum amount of water was added. The mixture was filtered through celite. The celite was washed with EtOAc. The organic layer was separated and dried (MgSO)₄) Filtered and the solvent evaporated. The residue (1.1g) was purified by column chromatography over silica gel (15-40 μm) (eluent: DCM/MeOH/NH)₄OH 93/7/0.5). The pure fractions were collected and the solvent was evaporated. The residue (0.25g) was crystallized from acetonitrile/diethyl ether. The precipitate was filtered and dried, yielding 0.11g (12%) of compound 11, m.p. 122 ℃.

Example B12

Preparation of Compound 86

A mixture of 4-chloro-2-pyridinecarbonitrile (154mg, 0.0011mol), intermediate 2(280mg, 0.0011mol) and 5N hydrochloric acid solution in 2-propanol (0.19ml, 0.0011mol) in DMF (2ml) was stirred at 100 ℃ under argon for 24 h, then cooled to room temperature and poured into water. The resulting mixture was made basic with saturated sodium bicarbonate and extracted 2 times with EtOAc. The organic layer was washed successively with a saturated solution of sodium bicarbonate and with brine, dried (MgSO)₄) Filtered and the solvent evaporated. The residue was purified by column chromatography over silica gel (40-63 μm) (eluent: EtOAc/cyclohexane 50/50). The pure fractions were collected and the solvent was evaporated, yielding 130mg (33%) of compound 86 as a beige foam.

Example B13

Preparation of Compound 87

A mixture of compound 86(110mg, 0.00031mol), sodium azide (22mg, 0.00034mol) and zinc bromide (70mg, 0.00031mol) in water (1ml) and 2-propanol (0.25ml) was stirred at 105 ℃ for 22 hours and then cooled to room temperature. 0.25N sodium hydroxide solution (3ml) was added and the mixture was stirred at room temperature for 1 hour. The precipitate was filtered and washed with MeOH, THF, and 1-butanol. The organic layer was evaporated and the solid formed was washed with MeOH and dried to yield 26mg (21%) of compound 87 as a beige solid, m.p. 210 ℃.

Example B14

Preparation of Compound 88

Following a procedure analogous to compound 86 (method B12), starting from intermediate 14(254mg, 0.00076mol) and intermediate 2(191mg, 0.00076 mol). After work-up, the residue was purified by column chromatography over silica gel (40-63 μm) (eluent: DCM/MeOH/NH)₄OH 95/5/0.2). The pure fractions were collected and the solvent was evaporated, yielding 139mg (30%) of compound 88 as a grey foam.

Example B15

Preparation of Compound 89

A mixture of compound 88(112mg, 0.00020mol) and palladium on carbon (10 wt%) (43mg, 0.000040mol) in MeOH (1ml) and EtOH (4ml) was stirred at room temperature under 1 atmosphere of hydrogen for 26 h. After filtration through celite, the solvent was evaporated and the residue was purified by SCX column chromatography. The pure fractions were collected and the solvent was evaporated, yielding 27mg (33%) of compound 89 as a grey foam.

Example B16

Preparation of Compound 90

Following a procedure analogous to compound 86 (method B12), starting from intermediate 15(850mg, 0.0024mol) and intermediate 2(561mg, 0.0022mol), the mixture was heated in a Biotage Initiator microwave apparatus at 120 ℃ for 2 hours. After work-up, the residue was purified by column chromatography over silica gel (40-63 μm) (eluent: DCM/MeOH 95/5). The pure fractions were collected and the solvent was evaporated, yielding 680mg (56%) of compound 90 as a brown foam.

Example B17

Preparation of Compound 91

Compound 90(200mg, 0.00036mol) was dissolved in EtOH (10ml) and MeOH (10 ml). Palladium on carbon (10 wt%) (100mg) was added. The mixture was stirred at room temperature in hydrogen for 24 hours. The mixture was filtered over celite and washed with MeOH. The solvent was evaporated, yielding 140mg (92%) of compound 91 as a green oil.

Example B18

Preparation of Compound 92

Following a procedure similar to compound 86, starting from intermediate 90(150mg, 0.00047mol) and intermediate 2(107mg, 0.00042mol), the mixture was heated in a Biotage Initiator microwave apparatus at 120 ℃ for 80 minutes. After work-up, the residue was purified by column chromatography over silica gel (40-63 μm) (eluent: DCM/MeOH 95/5). The pure fractions were collected and the solvent was evaporated, yielding 30mg (14%) of intermediate 90 as a green oil.

Example B19

Preparation of Compound 93

Compound 92(23mg, 0.000043mol) was dissolved in EtOH (1ml) and MeOH (1 ml). Palladium on carbon (10 wt%) (10mg) was added. The mixture was stirred at room temperature in hydrogen for 20 hours. The mixture was filtered over celite and washed with MeOH. The solvent was evaporated. The residue was purified by SCX column chromatography to yield 18mg (100%) of compound 93 as a green oil.

Example B20

Preparation of Compound 94

Following a procedure similar to compound 86, starting from intermediate 17(200mg, 0.00056mol) and intermediate 2(142mg, 0.00056mol), the mixture was heated in a Biotage Initiator microwave apparatus at 120 ℃ for 50 minutes. After work-up, the residue was passed through silica gel (40-63 μm) (eluent: DCM/MeOH/NH)₄OH 85/15/1) was purified by column chromatography. The pure fractions were collected and the solvent was evaporated, yielding 35mg (11%) of compound 94 as a red oil.

Example B21

Preparation of Compound 95

Compound 94(50mg, 0.000088mol) was dissolved in MeOH (3 ml). 5N hydrochloric acid solution added in 2-propanol (5ml). The mixture was stirred at room temperature for 17 hours. The solvent was evaporated. The residue was poured onto water and extracted with EtOAc. The organic layer was separated, washed with a saturated solution of sodium bicarbonate and dried (MgSO)₄) Filtration and evaporation of the solvent gave 15mg (36%) of compound 95 as a yellow oil.

Example B22

Preparation of Compound 96

Following a procedure similar to compound 86, starting from intermediate 18(160mg, 0.00070mol) and intermediate 2(160mg, 0.00064mol), the mixture was heated in a Biotage Initiator microwave apparatus at 120 ℃ for 1 hour. After work-up, the residue was purified by column chromatography over silica gel (40-63 μm) (eluent: DCM/MeOH/NH)₄OH 85/15/1). The pure fractions were collected and the solvent was evaporated, yielding 100mg (35%) of compound 96 as a green oil.

Example B23

Preparation of Compound 97

Compound 96(50mg, 0.00011mol) was dissolved in THF (3 ml). Lithium hydroxide (33mg, 0.00079mol) and water (1 drop) were added. The mixture was stirred at room temperature for 24 hours. The residue was poured onto water and extracted with EtOAc. The organic layer was separated and washed with 4N sodium hydroxide solution. The organic layer was separated, washed with 3N hydrochloric acid solution and dried (MgSO)₄) Filtered and the solvent evaporated. The residue was purified by SCX column chromatography to yield 20mg (41%) of compound 97 asA green oil.

Example B24

Preparation of Compound 98

Following a procedure similar to compound 86, starting from intermediate 19(170mg, 0.00079mol) and intermediate 2(180mg, 0.00071mol), the mixture was heated in a Biotage Initiator microwave apparatus at 120 ℃ for 80 minutes. After work-up, the residue was purified by column chromatography over silica gel (40-63 μm) (eluent: DCM/MeOH 85/15). The pure fractions were collected and the solvent was evaporated, yielding 224mg (73%) of compound 98 as a brown oil.

Example B25

Preparation of Compound 99

Compound 98(100mg, 0.00023mol) was dissolved in MeOH (5ml) and cooled at 0 deg.C. Sodium borohydride (27mg, 0.00069mol) was added slowly. The mixture was stirred at 80 ℃ for 4 hours. The reaction was quenched with water and the solvent was evaporated. The residue was extracted with EtOAc. The organic layer was separated, washed with a saturated solution of sodium bicarbonate and dried (MgSO)₄) Filtered and the solvent evaporated. The residue was purified by column chromatography over silica gel (40-63 μm) (eluent: DCM/MeOH 85/15). The pure fractions were collected and the solvent was evaporated, yielding 40mg (43%) of compound 99 as a colorless oil.

Example B26

Preparation of Compound 100

A mixture of intermediate 20(107mg, 0.00047mol), intermediate 2(118mg, 0.00047mol) and a 5N solution of hydrochloric acid in 2-propanol (0.12ml, 0.00072mol) in 1-methyl-2-pyrrolidone (2.3ml) was stirred at 120 ℃ under argon for 2 h, then cooled to room temperature and poured into water. The resulting mixture was basified with saturated sodium bicarbonate solution and extracted 3 times with EtOAc. The organic layer was separated, washed with water and brine, and dried (MgSO)₄) Filtered and the solvent evaporated. The residue was purified by column chromatography over silica gel (40-63 μm) (eluent: DCM/MeOH/NH)₄OH 90/10/0.5). The pure fractions were collected and the solvent was evaporated, yielding 61mg (26%) of compound 100 as a beige foam.

Example B27

Preparation of Compound 101

Lithium aluminium hydride (6.5mg, 0.00017mol) was added to a mixture of intermediate 21(53mg, 0.00017mol) in THF (1ml) at 0 ℃ under argon. The mixture was stirred at 0 ℃ for 1 hour, quenched with 5% potassium hydrogen sulfate solution, and extracted with EtOAc. The organic layer was separated and dried (MgSO)₄) Filtered and the solvent evaporated. The residue was dissolved in MeOH (1ml) and added dropwise to a mixture of N-4-pyridine-1, 4-phenylenediamine (32mg, 0.00017mol), sodium cyanoborohydride (16mg, 0.0025mol) and acetic acid (1 drop) in MeOH (0.5 ml). The mixture was stirred at room temperature for 20 hours, poured into water and extracted 2 times with EtOAc. The organic layer was separated, washed with a saturated solution of sodium bicarbonate and with brine, and dried (MgSO)₄) Filtering andthe solvent was evaporated. The residue was purified by column chromatography over silica gel (40-63 μm) (eluent: EtOAc/MeOH/NH)₄OH 90/10/0.3). The pure fractions were collected and the solvent was evaporated, yielding 25mg (34%) of compound 101 as a beige foam.

Example B28

Preparation of Compound 102

A mixture of intermediate 23(500mg, 0.0011mol), 2-chloro-4-bromopyridine (213mg, 0.0011mol), 4, 5-bis (diphenylphosphino) -9, 9-dimethylxanthene (83mg, 0.00014mol), sodium tert-butoxide (264mg, 0.0028mol) in toluene (7.5ml) was degassed under argon for 15 minutes. Tris (dibenzylideneacetone) dipalladium (0) -chloroform adduct (46mg, 0.000044mol) was added. The mixture was heated in a Biotage Initiator microwave apparatus at 100 ℃ for 90 seconds. The mixture was cooled to room temperature, then poured into water and extracted 2 times with EtOAc. The organic layer was washed 2 times with water, 1 time with brine, and dried (MgSO)₄) Filtered and the solvent evaporated. The residue was purified by column chromatography over silica gel (40-63 μm) (eluent: EtOAc/cyclohexane 30/70). The pure fractions were collected and the solvent was evaporated, yielding 254mg (41%) of compound 102 as a beige foam.

Example B29

Preparation of Compound 103

Compound 102(70mg, 0.00012mol) was dissolved in a 5N hydrochloric acid solution in 2-propanol (1.5 ml). Water (2 drops) was added. At room temperatureThe reaction mixture was stirred for 5 hours. The reaction was quenched and basified with saturated sodium bicarbonate solution and extracted 3 times with EtOAc. The organic layer was dried (MgSO)₄) Filtered and the solvent evaporated. The residue was purified by column chromatography over silica gel (40-63 μm) (eluent: EtOAc/cyclohexane 50/50). The pure fractions were collected and the solvent was evaporated, yielding 33mg (76%) of compound 103 as a white foam.

Example B30

Preparation of Compound 104

A mixture of 4-chloro- α -methyl-2-pyridinemethanol (170mg, 0.0011mol) and intermediate 2(271mg, 0.0011mol) in acetic acid (2ml) was stirred at 120 ℃ under argon for 1 hour, then allowed to cool to room temperature and poured onto water. The resulting mixture was basified with 4N sodium hydroxide solution and extracted 2 times with EtOAc. The organic layer was washed successively with saturated sodium bicarbonate solution and brine, dried (MgSO)₄) Filtered and the solvent evaporated. The residue was purified by column chromatography over silica gel (40-63 μm) (eluent: EtOAc/MeOH 80/20). The pure fractions were collected and the solvent was evaporated, yielding 190mg (47%) of compound 104 as a beige foam.

Example B31

Preparation of Compound 105

A mixture of compound 104(106mg, 0.00029mol) and activated manganese oxide (148mg, 0.0017mol) in chloroform (4ml) was stirred at room temperature for 6 hours. After filtration through a pad of celite, the solvent was evaporated and the residue was purified by column chromatography over silica gel (40-63 μm) (eluent: EtOAc). The pure fractions were collected and the solvent was evaporated, yielding 4mg (3%) of compound 105 as an orange solid.

Example B32

Preparation of Compound 106

Acetomidoxime (11mg, 0.00016mol) was added to activated under argon at room temperatureMolecular sieves and sodium hydride (3.72mg, 0.00016mol) in THF (0.6 ml). The reaction mixture was stirred at 70 ℃ for 1.5 hours and then cooled to room temperature. A solution of compound 200(50mg, 0.00013mol) in THF (0.6ml) was added. The reaction mixture was stirred at 70 ℃ for 1 hour. The reaction was quenched by addition of water and extracted 2 times with EtOAc. The organic layer was dried (MgSO₄) Filtered and the solvent evaporated. The residue was purified by column chromatography over silica gel (40-63 μm) (eluent: EtOAc/cyclohexane 70/30). The pure fractions were collected and the solvent was evaporated, yielding 16mg (30%) of compound 106 as a yellow oil.

Example B33

Preparation of Compound 107

Following a procedure analogous to compound 106, starting from benzylaminoxime (38mg, 0.00028mol) and compound 200(90mg, 0.00023mol), the residue was purified by column chromatography over silica gel (40-63 μm) after work-up (eluent: DCM/EtOAc 90/10). The pure fractions were collected and the solvent was evaporated, yielding 13mg (12%) of compound 107 as a yellow oil, m.p. 170 ℃ -174 ℃.

Example B34

Preparation of Compound 108

Following a similar procedure to compound 106, starting from N' -hydroxy-2-phenylacetimide amide (42mg, 0.00028mol) and compound 200(90mg, 0.00023 mol). After work-up, the residue was purified by column chromatography over silica gel (40-63 μm) (eluent: EtOAc/cyclohexane 50/50). The pure fractions were collected and the solvent was evaporated, yielding 50mg (45%) of compound 108 as a yellow solid, melting point 159 ℃ -161 ℃.

Example B35

Preparation of Compound 109

Following a procedure analogous to compound 86, starting from dimethyl 4-chloro-2, 6-pyridinedicarboxylate (228mg, 0.00099mol) and intermediate 2(250mg, 0.00099mol), the mixture was heated in a biotageinitor microwave apparatus at 120 ℃ for 2 hours. After work-up, the residue was purified by column chromatography over silica gel (40-63 μm) (eluent: acetone/cyclohexane 30/70 to 60/40). The pure fractions were collected and the solvent was evaporated, yielding 30mg (7%) of compound 109 as a yellow foam.

Example B36

Preparation of Compound 110

A mixture of intermediate 27(0.0016mol) and intermediate 2(0.0018mol) in acetic acid (35ml) was stirred in a CEM Discover microwave oven (P ═ 300W) at 120 ℃ for 5 minutes and then allowed to return to room temperature. Ice and sodium hydroxide were added. The mixture was filtered through celite. The celite was washed with DCM/MeOH (95/5). The organic layer was separated and dried (MgSO)₄) Filtered and the solvent evaporated. The residue (1g) was purified by column chromatography over kromasil (15-40 μm) (eluent: DCM/MeOH/NH)₄OH 95/5/0.5). The pure fractions were collected and the solvent was evaporated. From CH₃CN/MeOH crystallized the residue (0.3 g). The precipitate was filtered and dried, yielding 0.182g (29%) of compound 110, m.p. 136 ℃.

Example B37

Preparation of Compound 111

A mixture of intermediate 31(0.0016mol) and intermediate 2(0.0018mol) in acetic acid (3.5ml) was stirred in a CEM Discover microwave oven (P ═ 300W) at 120 ℃ for 5 minutes and then allowed to return to room temperature. Ice and concentrated NaOH were added. The mixture was extracted 2 times with DCM. The organic layer was separated and dried (MgSO)₄) Filtered and the solvent evaporated. The residue (0.9g) was purified by column chromatography over kromasil (10 μm) (eluent: DCM/MeOH/NH)₄OH 93/7/0.5). The pure fractions were collected and the solvent was evaporated. From CH₃The residue was crystallized from CN/MeOH/acetone (0.2 g). The precipitate was filtered and dried to yield 0.137g (21%) of compound 111, meltPoint 104 ℃.

Example B38

Preparation of Compound 112

A mixture of intermediate 33(0.0025mol) and intermediate 27(0.0025mol) in acetic acid (2.7ml) was stirred in a CEM Discover microwave oven (P ═ 300W) at 118 ℃ for 10 minutes and then allowed to return to room temperature. Water and 3N sodium hydroxide were added. The mixture was extracted with DCM. The organic layer was separated and dried (MgSO)₄) Filtered and the solvent evaporated. The residue (1.42g) was purified by column chromatography over silica gel (10 μm) (eluent: DCM/MeOH/NH)₄OH 95/5/0.5). The pure fractions were collected and the solvent was evaporated. The residue (0.56g) was taken up in 2-propanone/CH₃In CN. The precipitate was filtered and dried, yielding 0.506g (35%) of compound 112, m.p. 194 ℃.

Example B39

Preparation of Compound 113

Lithium borohydride (0.0026mol), then MeOH (1ml) was added portionwise to a solution of intermediate 35(0.0002mol) in THF (15ml) at 0 ℃ in a stream of nitrogen. The mixture was stirred at 0 ℃ for 4 hours and lithium borohydride (15 equivalents) was added. The mixture was stirred at room temperature overnight. Lithium borohydride (10 equivalents) was added. The mixture was stirred at room temperature for 4 hours and 30 minutes. Lithium borohydride (15 equivalents) was added. The mixture was stirred at room temperature for 24 hours and poured into water. MeOH and THF were evaporated. DCM was added. The mixture was filtered to give 0.01g of a first crude productA compound (I) is provided. The filtrate was extracted with DCM. The organic layer was separated and dried (MgSO)₄) Filtration and evaporation of the solvent gave 0.035g of the second crude product. Both fractions were purified by column chromatography over kromasil (5 μm) (eluent: DCM/MeOH/NH)₄OH 95/5/0.5 to 85/15/1.5). The pure fractions were collected and the solvent was evaporated, yielding 0.056g (28%) of compound 113, m.p. 154 ℃.

Example B41

Preparation of Compound 36

A mixture of 4-bromo-pyridine hydrochloride (0.034mol) and intermediate 2(0.0374mol) in acetic acid (13ml) was stirred at 110 ℃ for 40 minutes, then allowed to cool to room temperature, poured into ice water and basified with potassium carbonate. DCM was added. The mixture was stirred for 30 minutes and then filtered through celite. The filtrate was decanted. The celite was absorbed in DCM/MeOH (95/5). The mixture was stirred for 30 minutes and then filtered. The two filtrates were combined and dried (MgSO)₄) Filtered and the solvent evaporated. The residue (16.8g) was purified by column chromatography over silica gel (20-45 μm) (eluent: DCM/MeOH/NH)₄OH 92/8/0.5). The pure fractions were collected and the solvent was evaporated. The residue (4.2g) was taken up in 2-propanone. The precipitate was filtered and dried, yielding 3.6g (32%) of compound 36, mp 236 ℃.

Example B42

Preparation of Compound 115

N- (2-hydroxyethyl) piperazine (0.0019mol) and EDC (0.00 mol)19mol), HOBT (0.0019mol) and triethylamine (0.0019mol) were added to a solution of intermediate 41(0.0013mol) in DCM/DMF 75/25(20 ml). The mixture was stirred at room temperature overnight. 10% potassium carbonate was added. The mixture was extracted with DCM. The organic layer was separated and dried (MgSO)₄) Filtered and the solvent evaporated. The residue (0.32g) was purified by column chromatography over kromasil (10 μm) (eluent: DCM/MeOH/NH)₄OH 90/10/1). The pure fractions were collected and the solvent was evaporated, yielding 0.027g (4%) of compound 115.

Example B43

Preparation of Compound 116

1- [2- (2-hydroxyethoxy) ethyl]Piperazine (0.0019mol), EDCI (0.0019mol), HOBT (0.0019mol) and triethylamine (0.0019mol) were added to a solution of intermediate 41(0.0013mol) in DCM/DMF 75/25(20 ml). The mixture was stirred at room temperature overnight. 10% potassium carbonate was added. The mixture was extracted with DCM. The organic layer was separated and dried (MgSO)₄) Filtered and the solvent evaporated. The residue (0.38g) was purified by column chromatography over kromasil (10 μm) (eluent: DCM/MeOH/NH)₄OH 90/10/1). The pure fractions were collected and the solvent was evaporated, yielding 0.108g (16%) of compound 116.

Example B44

Preparation of Compound 117

Intermediate 2(0.002mol) and 4-chloro-1H-pyrrole were stirred in a CEM Discover microwave oven at 140 deg.C[2，3-d]A mixture of pyrimidine (0.002mol) in acetic acid (5ml) was used for 15 minutes. Acetic acid was evaporated. The crude product was dissolved in DCM. The organic layer was separated and dried (MgSO)₄) Filtered and the solvent evaporated. The residue (1g) was purified by column chromatography over silica gel (15-40 μm) (eluent: DCM/MeOH/NH)₄OH 93/7/0.5). The pure fractions were collected and the solvent was evaporated. The residue (0.27g) was crystallized from acetonitrile. The precipitate was filtered and dried, yielding 0.233g (31%) of compound 117, mp 211 ℃.

Example B45

Preparation of Compound 118

5-amino-1-pentanol (0.0019mol), EDC (0.0019mol), HOBT (0.0019mol) and triethylamine (0.0019mol) were added to a solution of intermediate 41(0.0013mol) in DCM/DMF 75/25(10 ml). The mixture was stirred at room temperature overnight. 10% potassium carbonate was added. The mixture was extracted with DCM. The organic layer was separated and dried (MgSO)₄) Filtered and the solvent evaporated. The residue (0.38g) was purified by column chromatography over kromasil (10 μm) (eluent: DCM/MeOH/NH)₄OH 90/10/1 to 80/20/2). The pure fractions were collected and the solvent was evaporated, yielding 0.128g of compound 118.

Example B46

Preparation of Compound 119

A mixture of intermediates 86 and 87(179mg, 0.00045mol) and 10% palladium on carbon (20mg) in EtOH was stirred at room temperature under 1 atmosphere of hydrogen for 16 h. After filtration through a pad of celite, the solvent was evaporated and the residue was purified by column chromatography over silica gel (40-63 μm) (eluent: DCM/MeOH 9/1). The pure fractions were collected and the solvent was evaporated, yielding 70mg (47%) of compound 119 as a yellow foam.

Example B47

Preparation of Compound 120

A mixture of intermediate 2(0.050mg, 0.000199mol), 4-quinolinecarboxaldehyde (31mg, 0.000199mol) in MeOH was stirred and refluxed overnight, then allowed to cool to room temperature. Sodium borohydride was added portionwise and the mixture was stirred at room temperature for 1 hour, hydrolyzed with water, extracted with DCM and passed over MgSO₄Dried and concentrated. The residue was purified by column chromatography over kromasil (10 μm) (eluent: DCM/MeOH/NH)₄OH 90/10/1 to 80/20/2). The pure fractions were collected and the solvent was evaporated, yielding 0.045g (45%) of compound 120.

Example B48

Preparation of Compound 121

A mixture of 3-pyridinecarboxaldehyde (0.0028mol) and intermediate 2(0.0028mol) in MeOH (20ml) was stirred and refluxed overnight before being allowed to return to room temperature. Sodium borohydride (0.0028mol) was added portionwise. The mixture was stirred at room temperature for 5 hours, and ice and water were added. The mixture was extracted with DCM. The organic layer was separated and dried (MgSO)₄) Filtered and the solvent evaporated. The residue was purified by column chromatography over silica gel (15-40 μm) (eluent: DCM/MeOH/NH)₄OH97/3/0.2)。The pure fractions were collected and the solvent was evaporated. The residue (0.4g) was taken up in HCl/isopropanol/diethyl ether. The precipitate was filtered and dried. Ice and water were added. The mixture was basified with 3N sodium hydroxide. The mixture was extracted with DCM. The residue (0.3g) was purified by column chromatography over silica gel (15-40 μm) (eluent: toluene/isopropanol/NH)₄OH 90/10/0.5). The pure fractions were collected and the solvent was evaporated. The residue (0.24g) was taken up in isopropanol/HCl/isopropanol/diethyl ether. The precipitate was filtered and dried to yield 0.23g (20%) of compound 121, which was isolated as the hydrochloride salt, mp 130 ℃.

Table F-1 lists compounds prepared according to one of the above examples. The following abbreviations are used in the tables: c. C₂HF₃O₂Represents trifluoroacetate, int. represents intermediate, comp. represents compound,. HCl represents hydrochloride, mp. represents melting point, ms. represents mass spectrum.

TABLE F-1

C. Pharmacological examples:

u87MG cells were human glioblastoma cells with wild-type p 53. In this cell line, MDM2 closely controls p53 expression.

The ability of compounds to preserve p53 in U87MG cells was determined by p53 enzyme linked immunosorbent assay. The p53 assay is a "sandwich" enzyme immunoassay using two polyclonal antibodies. Several antibodies specific for the p53 protein were immobilized on the surface of plastic wells. Any p53 present in the sample to be analyzed will bind to the capture antibody. The biotinylated detector polyclonal antibody also recognizes the p53 protein and will bind to any p53, which is held by the capture antibody. In turn, the detector antibody is bound by horseradish peroxidase-conjugated avidin. Horseradish peroxidase catalyzes the conversion of the chromogenic substrate o-phenylenediamine, the intensity of which is proportional to the amount of p53 protein bound to the plate. The colored reaction product is quantified using a spectrophotometer. Quantification was performed by constructing a standard curve using known concentrations of purified recombinant HIS-tagged p53 protein (see example c.1.).

The cellular activity of the compounds of the formula (I) was determined on U87MG tumor cells by colorimetric assay for cytotoxicity or viability (see example C.2).

C.1.p53 ELISA

At 37 deg.C, 5% CO in wet₂In the incubator of (1), Dulbecco's Minimal Essential Medium (DMEM) supplemented with 10% Fetal Calf Serum (FCS), 2mM L-glutamine, 1mM sodium pyruvate, 1.5g/L sodium bicarbonate and gentamicin was culturedU87MG cells (ATCC).

U87MG cells were seeded at 30.000 cells per well in 96-well plates, cultured for 24 hours, and treated with compound in a humidified incubator at 37 ℃ for 16 hours. After incubation, cells were washed 1 time with phosphate buffered saline and 30 μ l of low salt RIPA buffer (20mM tris, pH 7.0, 0.5mM EDTA, 1% Nonidet P40, 0.5% DOC, 0.05% SDS, 1mM PMSF, 1 μ g/ml aprotinin and 0.5 μ/ml leupeptin) was added per well. The plate was placed on ice for 30 minutes to complete lysis. P53 protein was detected in the lysates by using the sandwich ELISA described below.

To coat with buffer (0.1M NaHCO)₃pH8.2) was coated with 50. mu.l per well of a high-binding polystyrene EIA/RIA 96-well plate (Costar 9018) with a capture antibody pAb122(Roche 1413147) at a concentration of 2. mu.g/ml. The antibody was allowed to adhere overnight at 4 ℃. The coated plates were washed 1 time with Phosphate Buffered Saline (PBS)/0.05% Tween20 and incubated for 2 hours at room temperature with the addition of 300. mu.l of blocking buffer (PBS, 1% Bovine Serum Albumin (BSA)). Dilutions (ranging from 3 to 200ng/ml) of purified recombinant HIS-tagged p53 protein were prepared in blocking buffer and used as standards.

The plates were washed 2 times with PBS/0.05% Tween20 and blocking buffer or standard was added, 80. mu.l per well. Add 20. mu.l of lysis buffer to the standard. Samples were added to other wells, 20. mu.l of lysate per well. After overnight incubation at 4 ℃, the plates were washed 2 times with PBS/0.05% Tween 20. An aliquot of 100. mu.l of secondary polyclonal antibody p53(FL-393) (Tebubio, sc-6243) was added to each well at a concentration of 1. mu.g/ml in blocking buffer and allowed to adhere for 2 hours at room temperature. The plates were washed 3 times with PBS/0.05% Tween 20. The detection antibody anti-rabbit HRP (sc-2004, Tebubio) was added at 0.04. mu.g/ml in PBS/1% BSA and incubated for 1 hour at room temperature. The plates were washed 3 times with PBS/0.05% Tween20 and 100. mu.l of matrix buffer (matrix buffer was added shortly before use by adding 1 plate of 10mg of o-phenylenediamine (OPD) from Sigma and 125. mu.l of 3% H₂O₂To 25ml OPD buffer: 35mM citric acid, 132mM Na₂HPO₄Prepared at pH 5.6). After 5 to 10 minutes, 50. mu.l of stop buffer (1M H) was added per well₂SO₄) The color reaction was terminated. The absorbance at the two wavelength of 450/655nm was measured using a Biorad microplate reader and the results were then analyzed.

For each experiment, a control (no drug) and a blank culture (no cells or drug) were run in parallel. Blank values were subtracted from all control and sample values. For each sample, the value of p53 (in absorbance units) is expressed as a percentage of the p53 value present in the control group. Percent retention above 130% is defined as significant. Here, the effect of the test compound is expressed as the lowest dose (LAD) that provides a p53 value of at least 130% present in the control group (see Table F-2).

C.2. Proliferation assay

All compounds tested were dissolved in DMSO and additional dilutions were made in culture medium. The final DMSO concentration never exceeded 0.1% (v/v) in the cell proliferation assay. The control group contained U87MG cells and DMSO but no compound, while the blank group contained DMSO but no cells.

U87MG cells were seeded at 3000 cells/well/100. mu.l in 96-well cell culture dishes. After 24 hours, the medium was replaced and the compound and/or solvent was added to a final volume of 200 μ l. After 4 days of culture, media was replaced with 200 μ l of fresh media and cell growth was assessed using an MTT-based assay. Therefore, 25 μ l of MTT solution (0.5% MTT in phosphate-buffered saline, research grade, from Serva) was added to each well and the cells were further cultured for 2 hours at 37 ℃. The medium was then carefully aspirated and the blue MTT-formazan was dissolved by adding 25. mu.l of 0.1M glycine and 100. mu.l DMSO to each wellAnd (3) obtaining the product. The plate was shaken on a microplate shaker for a further 10 minutes before the absorbance was read at 540nm by a microplate reader.

In the experiment, the results for each experimental condition were averaged over 3 replicate wells. For initial screening purposes, at a single fixed concentration of 10^-5M test compound. For the active compound, the experiment was repeated to establish the entire concentration-response curve. For each experiment, a control (no drug) and a blank culture (no cells or drug) were run in parallel. Blank values were subtracted from all control and sample values. For each sample, the mean value of cell growth (absorbance units) is expressed as a percentage of the mean value of cell growth of the control group. When appropriate, the IC is calculated using probabilistic Unit analysis (probit analysis) of the hierarchical data₅₀Values (concentration of drug required to reduce cell growth to 50% of control) (Finney, d.j., Probit analytes, 2 nd edition, chapter 10, GradedResponses, Cambridge University Press, Cambridge 1962). The effect of the test compounds is expressed herein as pIC₅₀(IC₅₀Negative logarithm of value) (see table F-2).

TABLE F-2: table F-2 shows the results for the compounds tested according to examples C.1 and C.2

Co No	p53-elisaLAD	Cell proliferation pIC50
			1	3.0E-08	＞8.0
2	3.0E-07	7.2
			3	＞1.0E-05	5.3
4	3.0E-08	8.0
			5	3.0E-08
6	＞1.0E-05	5.5
			7	1.0E-05	5.7
8	＞1.0E-05	5.3
			9	＞1.0E-05
10	＞1.0E-05	5.9
			11	＞1.0E-05	5.3
12	3.0E-07	7.9
			13	1.0E-07	7.6

Co No	p53-elisaLAD	Cell proliferation pIC50
			14	3.0E-07	7.4
15	＞1.0E-05	7.3
			16	＞1.0E-05	7.4
17	＞1.0E-05	6.2
			18	1.0E-07	6.3
19	3.0E-07	6.7
			20	3.0E-07	7.0
21	3.0E-08	8.0
			22	1.0E-07	7.7
23	1.0E-06	6.4
			24	1.0E-07	＞8.0
25	＞1.0E-05	7.4
			26	3.0E-06	7.0
27	3.0E-06	7.1
			28	＞1.0E-05	6.7
29	3.0E-06	6.6
			30	＞1.0E-05	6.5
31	＞1.0E-05	5.9
			32	3.0E-06	6.8
33	＞1.0E-05	7.2
			34	＞1.0E-05	7.3
35	1.0E-06	7.4
			36	1.0E-06	6.7
37	3.0E-07	6.8
			38	＞1.0E-05
39	1.0E-05	6.2
			40	＞1.0E-05
41	＞1.0E-05
			42	＞1.0E-05
43	＞1.0E-05
			44	＞1.0E-05
45	＞1.0E-05	6.0
			46	1.0E-06	6.6
47	1.0E-05	6.8
			48	1.0E-05	6.8
49	＞1.0E-05	＜5.0
			50	3.0E-06	7.0
51	＞1.0E-05	6.5
			52	＞1.0E-05	6.3
53	＞1.0E-05	6.2
			54	1.0E-06	6.9
55	3.0E-07	6.3
			56	＞1.0E-05	5.6
57	＞1.0E-05	6.1

Co No	p53-elisaLAD	Cell proliferation pIC50
			58	＞1.0E-05	＜5.0
59	1.0E-06	6.4
			60	＞1.0E-05	7.0
61	＞1.0E-05	6.5
			62	＞1.0E-05	5.6
63	＞1.0E-05	5.8
			64	1.0E-06	6.4
65	＞1.0E-05	＜5.0
			66	3.0E-07	7.2
67	＞1.0E-05	5.9
			68	＞1.0E-05	5.6
69	1.0E-07	7.0
			70	＞1.0E-05	6.6
71	＞1.0E-05	6.1
			72	＞1.0E-05	5.7
73	＞1.0E-05	6.3
			74	＞1.0E-05	5.8
75	＞1.0E-05	5.5
			76	＞1.0E-05	＜5.0
77	＞1.0E-05	5.5
			78	＞1.0E-05	5.0
79	＞1.0E-05	5.6
			82	＞1.0E-05	＜5.0
83	＞1.0E-05	5.5
			84	＞1.0E-05	5.8
85	＞1.0E-05	6.8
			86	＞1.00E-05	＜5.0
87	＞1.00E-05	＜5.0
			88	＞1.00E-05	5.5
89	3.00E-06	5.4
			90	＞1.00E-05	5.6
91	＞1.00E-05	5.6
			92	＞1.00E-05	5.5
93	＞1.00E-05	＜5.0
			95	＞1.00E-05	5.1
96	＞1.00E-05	＜5.0
			97	1.00E-06	＜5.0
98	＞1.00E-05	5.4
			99	1.00E-05	5.6
100	＞1.00E-05	5.4
			101	＞1.00E-05	5.6
102	＞1.00E-05	＜5.0
			103	1.00E-05	5.4
104	3.00E-06	5.5

Co No	p53-elisaLAD	Cell proliferation pIC50
			105	＞1.00E-05	5.1
106	＞1.00E-05	5.8
			107	＞1.00E-05
108	＞1.00E-05
			109	1.00E-06	＜5.0
110	1.00E-07	8.0
			111	1.00E-07	7.1
112	1.00E-07	7.5
			113	＞1.00E-05	＜5.0
114	＞1.00E-05	＜5.0
			114	＞1.00E-05	＜5.0
115	＞1.00E-05	＜5.0
			116	＞1.00E-05	＜5.0
117	＞1.00E-05	＜5.0
			118	＞1.00E-05	＜5.0
119	＞1.00E-05	＜5.0
			120	＞1.00E-05	5.3
121	＞1.00E-05	＜5.0
			123		5.3
124		5.3
			125	＞1.00E-05	5.4
126	＞1.00E-05	＜5.0
			127	＞1.00E-05	5.1
128	＞1.00E-05	5.5
			129	3.00E-06	5.7
130	＞1.00E-05	5.8
			131	＞1.00E-05	5.6
132	＞1.00E-05	＜5.0
			134	＞1.00E-05	5.9
135	1.00E-06	6.0
			136	＞1.00E-05	5.7
137	＞1.00E-05	5.5
			138	＞1.00E-05	5.8
139	＞1.00E-05	5.7
			140	＞1.00E-05	5.6
141	1.00E-05	5.4
			142	3.00E-06	5.5
143	＞1.00E-05	5.5
			144		5.5
145		5.6
			146	＞1.00E-05	5.1
147	＞1.00E-05	5.3
			148	3.00E-07	5.5
149	＞1.00E-05	5.7

Co No	p53-elisaLAD	Cell proliferation pIC50
			150	1.00E-06	5.5
151	1.00E-06	＜5.0
			152	＞1.00E-05	5.0
153	＞1.00E-05	5.6
			154	＞1.00E-05	5.5
155	＞1.00E-05	5.7
			156	＞3.00E-06	5.5
157	＞1.00E-05	5.8
			158	＞1.00E-05	＜5.0
159	＞1.00E-05	5.5
			160	＞1.00E-05	6.4
161	＞1.00E-05	6.0
			162	＞1.00E-05	5.5
163	＞1.00E-05	5.5
			164	＞1.00E-05	5.6
165	3.00E-06	5.3
			166	＞1.00E-05	＜5.0
167	＞1.00E-05	5.4
			168	＞1.00E-05	5.7
169	＞1.00E-05	6.4
			170	3.00E-07	5.5
171	＞1.00E-05	＜5.0
			172	＞1.00E-05	＜5.0
173	＞1.00E-05	＜5.0
			174	＞1.00E-05
175	＞1.00E-05
			176	3.00E-06
177	3.00E-07	7.3
			178	＞1.00E-05	5.8
179	3.00E-06	6.6
			180	＞1.00E-05	6.2
181	3.00E-07	6.6
			182	＞1.00E-05	5.8
183	1.00E-05	6.3
			184	＞1.00E-05	6.0
185	3.00E-06	5.7
			186	1.00E-06	6.0
187	1.00E-06	6.4
			188	1.00E-06	6.1
189	＞1.00E-05	5.5
			190	＞1.00E-05	5.4
191	＞1.00E-05	5.5
			192	1.00E-06	＜5.0
193	3.00E-06	6.0

Co No	p53-elisaLAD	Cell proliferation pIC50
			194	＞1.00E-05	5.2
195	1.00E-06	7.1
			196	＞1.00E-05	6.7
197	1.00E-07	6.6
			198	1.00E-06	5.9
199	＞1.00E-05	5.7
			201	＞1.00E-05	5.5
202	＞1.00E-05	5.5
			203	＞1.00E-05	5.5
204	＞1.00E-05	5.1
			205	3.00E-06	6.1
206	＞1.00E-05	5.5
			207	＞1.00E-05	6.1
208	3.00E-06	＜5.0
			209	＞1.00E-05	＜5.0
210	＞1.00E-05	＜5.0
			211	3.00E-07	7.2
212	＞1.00E-05	5.8
			213	3.00E-06	＜5.0
214	1.00E-06	＜5.0
			215	＞1.00E-05	5.5
216	1.00E-06	5.6
			217	＞1.00E-05	5.4
218	＞1.00E-05	＜5.0
			219	＞1.00E-05	5.4
220	3.00E-06	5.2
			221	＞1.00E-05	5.4
222	＞1.00E-05	5.4
			223	＞1.00E-05	6.1
224	＞1.00E-05	5.4
			225	＞1.00E-05	6.8
226	3.00E-06	5.5
			227	＞1.00E-05	5.1
228	1.00E-06	5.1
			229	1.00E-07	7.0

D. Composition examples: film coated tablet

Preparation of tablet cores

A mixture of 100g of the compound of formula (I), 570g of lactose and 200g of starch is mixed thoroughly and then moistened with a solution of 5g of sodium lauryl sulfate and 10g of polyvinylpyrrolidone in about 200ml of water. The wet powder was sieved, dried and sieved again. Then 100g microcrystalline cellulose and 15g hydrogenated vegetable oil were added. The whole was mixed well and compressed into tablets to obtain 10.000 tablets each containing 10mg of the compound of formula (I).

Coating film

A solution of 5g of ethylcellulose in 150ml of dichloromethane is added to a solution of 10g of methylcellulose in 75ml of denatured ethanol. Then 75ml of dichloromethane and 2.5ml of 1, 2, 3-propanetriol are added, 10g of polyethylene glycol are melted and dissolved in 75ml of dichloromethane. The latter solution was added to the former, followed by 2.5g magnesium stearate, 5g polyvinylpyrrolidone and 30ml concentrated coloured suspension, and all homogenized. The tablet cores are coated in a coating device with the mixture thus obtained.

Claims (21)

1. A compound of the formula (I),

an addition salt or a stereochemically isomeric form thereof, wherein:

m is 0, 1 or 2, and when m is 0, it means a direct bond;

n is 0, 1, 2 or 3, and when n is 0, it means a direct bond;

p is 0 or 1, and when p is 0, it means a direct bond;

s is 0 or 1, and when s is 0, it means a direct bond;

t is 0 or 1, and when t is 0, it means a direct bond;

x is C (═ O) or CHR⁸(ii) a Wherein

R⁸Is hydrogen, C_1-6Alkyl radical, C_3-7Cycloalkyl, -C (═ O) -NR¹⁷R¹⁸Hydroxycarbonyl radical, aryl radical C_1-6Alkoxycarbonyl, heteroaryl, heteroarylcarbonyl, heteroaryl C_1-6Alkoxycarbonyl, piperazinylcarbonyl, pyrrolidinyl, piperidinylcarbonyl, C_1-6Alkoxycarbonyl, C substituted by a substituent selected from the group consisting of hydroxy, amino, aryl and heteroaryl_1-6An alkyl group; c substituted by a substituent selected from the group consisting of hydroxy, amino, aryl and heteroaryl_3-7A cycloalkyl group; by hydroxy, hydroxy C_1-6Alkyl, hydroxy C_1-6Alkoxy radical C_1-6An alkyl-substituted piperazinecarbonyl group; by hydroxy radicals C_1-6Alkyl-substituted pyrrolidinyl; or by one or two radicals selected from hydroxy, C_1-6Alkyl, hydroxy C_1-6Alkyl radical, C_{1 -6}Alkoxy radical C_1-6Alkyl radical, C_1-6Alkyl (dihydroxy) C_1-6Alkyl or C_1-6Alkoxy (hydroxy) C_{1 -6}Piperidinylcarbonyl substituted with a substituent for alkyl;

R¹⁷and R¹⁸Each independently selected from hydrogen and C_1-6Alkyl, di (C)_1-6Alkyl) amino C_1-6Alkyl, aryl C_1-6Alkyl radical, C_1-6Alkoxy radical C_1-6Alkyl, hydroxy C_1-6Alkyl, hydroxy C_1-6Alkyl radical (C)_1-6Alkyl) or hydroxy C_1-6Alkyl (aryl C)_1-6Alkyl groups);

is-CR⁹C < and the dotted line is a bond or-CHR⁹-CH <; wherein

Each R⁹Independently is hydrogen or C_1-6An alkyl group;

R¹is hydrogen, aryl, heteroaryl, C_1-6Alkoxycarbonyl group, C_1-12Alkyl or substituted by one or two groups independently selected from hydroxy, aryl, heteroaryl, amino, C_1-6Alkoxy, mono-or di (C)_{1 -6}Alkyl) amino, morpholinyl, piperidinyl, pyrrolidinyl, piperazinyl, C_1-6Alkyl piperazinyl, aryl C_1-6Alkyl piperazinyl, heteroaryl C_1-6Alkyl piperazinyl, C_3-7Cycloalkyl piperazinyl and C_3-7Cycloalkyl radical C_1-6C substituted by substituents of alkylpiperazino radicals_1-12An alkyl group;

R²is hydrogen, halogen, C_1-6Alkyl radical, C_1-6Alkoxy, aryl C_1-6Alkoxy, heteroaryl C_1-6Alkoxy, phenylthio, hydroxy C_1-6Alkylcarbonyl, C substituted by a substituent selected from amino, aryl and heteroaryl_1-6An alkyl group; or C substituted by a substituent selected from the group consisting of amino, aryl and heteroaryl_3-7A cycloalkyl group;

R³is hydrogen, C_1-6Alkyl, heteroaryl, C_3-7Cycloalkyl, C substituted by a substituent selected from the group consisting of hydroxy, amino, aryl and heteroaryl_1-6An alkyl group; or C substituted by a substituent selected from the group consisting of hydroxy, amino, aryl and heteroaryl_3-7A cycloalkyl group;

R⁴and R⁵Each independently of the others is hydrogen, halogen, C_1-6Alkyl, polyhalo C_1-6Alkyl, cyano C_1-6Alkyl, hydroxy, amino or C_1-6An alkoxy group; or

R⁴And R⁵Together optionally forming a divalent group selected from methylenedioxy or ethylenedioxy;

R⁶is hydrogen, C_1-6Alkoxycarbonyl or C_1-6An alkyl group;

when p is 1, then R⁷Is hydrogen, aryl C_1-6Alkyl, hydroxy or heteroaryl C_1-6An alkyl group;

z is a group selected from:

wherein

Each R¹⁰Or R¹¹Each independently selected from hydrogen, halogen, hydroxy, amino, C_1-6Alkyl, nitro, polyhalo C_1-6Alkyl, cyano C_1-6Alkyl, tetrazole C_1-6Alkyl, aryl, heteroaryl, aryl C_1-6Alkyl, heteroaryl C_1-6Alkyl, aryl (hydroxy) C_1-6Alkyl, heteroaryl (hydroxy) C_1-6Alkyl, arylcarbonyl, heteroarylcarbonyl, C_1-6Alkylcarbonyl, aryl C_1-6Alkylcarbonyl, heteroaryl C_1-6Alkylcarbonyl group, C_1-6Alkoxy radical, C_3-7Cycloalkyl carbonyl group, C_3-7Cycloalkyl (hydroxy) C_1-6Alkyl, aryl C_1-6Alkoxy radical C_1-6Alkyl radical, C_1-6Alkoxy radical C_1-6Alkoxy radical C_1-6Alkyl radical, C_1-6Alkylcarbonyloxy C_1-6Alkyl radical, C_1-6Alkoxycarbonyl radical C_1-6Alkoxy radical C_1-6Alkyl, hydroxy C_1-6Alkoxy radical C_1-6Alkyl radical, C_1-6Alkoxycarbonyl radical C_2-6Alkenyl radical, C_1-6Alkoxy radical C_1-6Alkyl radical, C_1-6Alkoxycarbonyl group, C_1-6Alkylcarbonyloxy, aminocarbonyl, hydroxy C_1-6Alkyl, amino C_1-6Alkyl, hydroxycarbonyl C_1-6Alkyl and- (CH)₂)_v-(C(＝O)_r)-(CHR¹⁹)_u-NR¹³R¹⁴(ii) a Wherein

v is 0, 1, 2, 3, 4, 5 or 6, and when v is 0, it means a direct bond;

r is 0 or 1, and when r is 0, it means a direct bond;

u is 0, 1, 2, 3, 4, 5 or 6, and when u is 0, it means a direct bond;

R¹⁹is hydrogen or C_1-6An alkyl group;

R¹²is hydrogen, C_1-6Alkyl radical, C_3-7Cycloalkyl, selected from hydroxy, amino, C_1-6C substituted by alkoxy and aryl substituents_1-6An alkyl group; or selected from hydroxy, amino, aryl and C_1-6C substituted by substituents of alkoxy_3-7A cycloalkyl group;

R¹³and R¹⁴Each independently selected from hydrogen and C_1-12Alkyl radical, C_1-6Alkylcarbonyl group, C_1-6Alkylsulfonyl, aryl C_1-6Alkylcarbonyl group, C_3-7Cycloalkyl radical, C_3-7Cycloalkyl carbonyl, - (CH)₂)_k-NR¹⁵R¹⁶Selected from hydroxy, hydroxycarbonyl, cyano, C_1-6Alkoxycarbonyl group, C_1-6C substituted by substituents of alkoxy, aryl or heteroaryl_1-12An alkyl group; or selected from hydroxy, C_1-6Alkoxy, aryl, amino, aryl C_1-6Alkyl, heteroaryl or heteroaryl C_{1- 6}C substituted by substituents of alkyl groups_3-7A cycloalkyl group; or

R¹³And R¹⁴Together with the nitrogen to which they are attached may optionally form morpholinyl, piperidinyl, pyrrolidinyl, piperazinyl or be selected from C_1-6Alkyl, aryl C_1-6Alkyl, aryl C_1-6Alkoxycarbonyl, heteroaryl C_1-6Alkyl radical, C_3-7Cycloalkyl and C_3-7Cycloalkyl radical C_1-6Piperazinyl substituted with a substituent for alkyl; wherein

k is 0, 1, 2, 3, 4, 5 or 6, and when k is 0, it means a direct bond;

R¹⁵and R¹⁶Each independently selected from hydrogen and C_1-6Alkyl, aryl C_1-6Alkoxycarbonyl group, C_3-7Cycloalkyl, selected from hydroxy, C_1-6C substituted by substituents of alkoxy, aryl and heteroaryl_1-12An alkyl group; and is selected from hydroxy, C_1-6Alkoxy, aryl C_1-6Alkyl, heteroaryl and heteroaryl C_1-6C substituted by substituents of alkyl groups_3-7A cycloalkyl group; or

R¹⁵And R¹⁶Optionally formed with the nitrogen to which they are attachedMorpholinyl, piperazinyl or by C_{1 -6}Piperazinyl substituted with alkoxycarbonyl;

aryl is phenyl or naphthyl;

each phenyl or naphthyl group may be optionally substituted with one, two or three substituents each independently selected from halogen, hydroxy, C_1-6Alkyl, amino, polyhalo C_1-6Alkyl and C_1-6An alkoxy group; and

each phenyl or naphthyl group may be optionally substituted with a divalent group selected from methylenedioxy or ethylenedioxy;

heteroaryl is pyridyl, indolyl, quinolyl, imidazolyl, furyl, thienyl,Oxadiazolyl, tetrazolyl, benzofuranyl or tetrahydrofuranyl;

each of pyridyl, indolyl, quinolyl, imidazolyl, furyl, thienyl,The oxadiazolyl, tetrazolyl, benzofuranyl or tetrahydrofuranyl group may be optionally substituted with one, two or three substituents each independently selected from halogen, hydroxy, C_1-6Alkyl, amino, polyhalo C_1-6Alkyl, aryl C_1-6Alkyl or C_1-6An alkoxy group; and

each pyridyl, indolyl, quinolinyl, imidazolyl, furyl, thienyl, benzofuryl or tetrahydrofuranyl group may be optionally substituted with a divalent group selected from methylenedioxy or ethylenedioxy;

with the following conditions:

when m is 1; by removal of R from the benzene ring²The outer substituents are in the meta position; s is 0; and t is 0, then

Z is a group selected from (a-1), (a-3), (a-4), (a-5), (a-6), (a-7), (a-8) or (a-9).

2. A compound according to claim 1, wherein:

R⁸is hydrogen, -C (═ O) -NR¹⁷R¹⁸Aryl radical C_1-6Alkoxycarbonyl, hydroxy-substituted C_1-6Alkyl, by hydroxy, hydroxy C_1-6Alkyl, hydroxy C_1-6Alkoxy radical C_1-6Alkyl-substituted piperazinecarbonyl, substituted by hydroxy C_1-6Pyrrolidinyl substituted by alkyl or by one or two selected from hydroxy, C_1-6Alkyl, hydroxy C_1-6Alkyl radical, C_1-6Alkoxy radical C_1-6Alkyl radical, C_1-6Alkyl (dihydroxy) C_1-6Alkyl or C_1-6Alkoxy (hydroxy) C_1-6Piperidinylcarbonyl substituted with a substituent for alkyl; r¹⁷And R¹⁸Each independently selected from hydrogen and C_1-6Alkyl, di (C)_1-6Alkyl) amino C_{1 -6}Alkyl, aryl C_1-6Alkyl radical, C_1-6Alkoxy radical C_1-6Alkyl or hydroxy C_1-6An alkyl group; r¹Is hydrogen, heteroaryl, C_1-6Alkoxycarbonyl group, C_1-23Alkyl or C substituted by heteroaryl_1-12An alkyl group; r²Is hydrogen, halogen, C_1-6Alkyl radical, C_1-6Alkoxy, aryl C_1-6Alkoxy or thiophenyl; r³Is hydrogen, C_1-6Alkyl or heteroaryl; r⁴And R⁵Each independently of the others is hydrogen, halogen, C_1-6Alkyl, cyano C_1-6Alkyl, hydroxy or C_1-6An alkoxy group; when p is 1, then R⁷Is aryl C_1-6Alkyl or hydroxy; z is a group selected from (a-1), (a-2), (a-3), (a-4), (a-5), (a-6), (a-8), (a-9), (a-10) and (a-11); each R¹⁰Or R¹¹Each independently selected from hydrogen, halogen, hydroxy, amino, C_1-6Alkyl, nitro, polyhalo C_1-6Alkyl, cyano C_1-6Alkyl, tetrazole C_1-6Alkyl, aryl, heteroaryl C_1-6Alkyl, aryl (hydroxy) C_1-6Alkyl, arylcarbonyl, C_1-6Alkylcarbonyl group, C_3-7Cycloalkyl carbonyl group, C_3-7Cycloalkyl (hydroxy) C_1-6Alkyl, aryl C_1-6Alkoxy radical C_1-6Alkyl radical, C_1-6Alkoxy radical C_1-6Alkoxy radical C_1-6Alkyl radical, C_1-6Alkylcarbonyloxy C_{1 -6}Alkyl radical, C_1-6Alkoxycarbonyl radical C_1-6Alkoxy radical C_1-6Alkyl, hydroxy C_1-6Alkoxy radical C_{1 -6}Alkyl radical, C_1-6Alkoxycarbonyl radical C_2-6Alkenyl radical, C_1-6Alkoxy radical C_1-6Alkyl radical, C_1-6Alkoxycarbonyl, aminocarbonyl, hydroxy C_1-6Alkyl, amino C_1-6Alkyl, hydroxycarbonyl C_1-6Alkyl and- (CH)₂)_v-(C(＝O)_r)-(CHR¹⁹)_u-NR¹³R¹⁴(ii) a v is 0 or 1; u is 0 or 1; r¹²Is hydrogen or C_1-6An alkyl group; r¹³And R¹⁴Each independently selected from hydrogen and C_1-12Alkyl radical, C_1-6Alkylcarbonyl group, C_1-6Alkylsulfonyl, aryl C_1-6Alkylcarbonyl group, C_3-7Cycloalkyl carbonyl, - (CH)₂)_k-NR¹⁵R¹⁶Selected from hydroxy, hydroxycarbonyl, cyano, C_1-6C substituted by substituents of alkoxycarbonyl or aryl groups_1-12An alkyl group; r¹³And R¹⁴Together with the nitrogen to which they are attached may optionally form morpholinyl, pyrrolidinyl, piperazinyl or be selected from C_1-6Alkyl or aryl radicals C_1-6Piperazinyl substituted with a substituent of alkoxycarbonyl; k is 2; r¹⁵And R¹⁶Each independently selected from hydrogen and C_1-6Alkyl or aryl radicals C_1-6An alkoxycarbonyl group; r¹⁵And R¹⁶Together with the nitrogen to which they are attached may optionally form morpholinyl or piperazinyl or be C_1-6Alkoxycarbonyl-substituted piperazinyl; aryl is phenyl or phenyl substituted by halogen; heteroaryl is pyridyl, indolyl,Oxadiazolyl or tetrazolyl; and each of the pyridyl, indolyl, etc,Oxadiazolyl or tetrazolyl may optionally be substituted with one selected from C_1-6Alkyl, aryl or aryl C_1-6Alkyl substituents.

3. A compound according to claim 1 or 2, wherein

m is 0; n is 1; p is 0; s is 0; t is 0; x is CHR⁸；R⁸Is hydrogen;is-CR⁹C <, and the dotted line is a bond; each R⁹Is hydrogen; r¹Is hydrogen; r²Is hydrogen or C_1-6An alkoxy group; r³Is hydrogen; r⁴And R⁵Each independently is hydrogen, C_1-6Alkyl or C_1-6An alkoxy group; r⁶Is hydrogen; z is a group selected from (a-1), (a-2), (a-3) or (a-4); and R¹⁰Or R¹¹Each independently selected from hydrogen, hydroxy or hydroxy C_1-6An alkyl group.

4. The compound according to claim 1, 2 or 3, wherein the compound is compound No. 1, compound No. 21, compound No. 4, compound No. 5, compound No. 36, compound No. 69, compound No. 110, compound No. 111, compound No. 112, compound No. 229 or compound No. 37:

5. the compound according to claim 4, wherein said compound is

6. A compound according to any one of claims 1 to 3, wherein m is 0; n is 1; p is 0; s is 0; t is 0.

7. A compound according to any one of claims 1 to 3, wherein X is CHR⁸Wherein R is⁸Is hydrogen.

8. A compound according to any one of claims 1 to 3, whereinis-CR⁹Wherein R <, C⁹Is hydrogen.

9. A compound according to any one of claims 1 to 3, wherein R¹Is hydrogen; r³Is hydrogen; r⁶Is hydrogen.

10. A compound according to any one of claims 1 to 3, wherein R⁴And R⁵Each independently is hydrogen, C_1-6Alkyl or C_1-6An alkoxy group.

11. A compound according to any one of claims 1 to 3, wherein Z is a group selected from (a-1), (a-2), (a-3) or (a-4).

12. A compound according to any one of claims 1 to 3, wherein R¹⁰And R¹¹Each independently selected from hydrogen, hydroxy or hydroxy C_1-6An alkyl group.

13. A compound according to any one of claims 1 to 3, wherein R²Is hydrogen or C_1-6An alkoxy group.

14. A compound according to any one of claims 1 to 3, wherein the compound is

In addition salt form.

15. The compound according to claim 14, wherein said compound is

Or a pharmaceutically acceptable salt thereof.

16. A compound according to claim 1, 2, 3, 4, 5 or 15 for use as a medicament.

17. A pharmaceutical composition comprising a pharmaceutically acceptable carrier and, as active ingredient, a therapeutically effective amount of a compound according to any one of claims 1 to 16.

18. A process for preparing a pharmaceutical composition according to claim 17, wherein a pharmaceutically acceptable carrier is intimately mixed with a compound according to any one of claims 1 to 16.

19. Use of a compound according to any one of claims 1 to 16 for the manufacture of a medicament for the treatment of a disorder mediated by a p53-MDM2 interaction.

20. A process for the preparation of a compound according to claim 1, characterized in that

a) Reacting an intermediate of formula (II) with an intermediate of formula (III) wherein W is a suitable leaving group,

wherein the variables are as defined in claim 1;

b) a compound of formula (I) wherein X is C (═ O), i.e. a compound of formula (I-b), is converted to a compound of formula (I-b) wherein X is CH, in the presence of lithium aluminium hydride in a suitable solvent₂The compound of formula (I), i.e. the compound of formula (I-a),

wherein the variables are as defined in claim 1;

c) reacting an appropriate formaldehyde compound of formula (IV) with an intermediate of formula (V) in the presence of an appropriate reagent in an appropriate solvent,

wherein the variables are as defined in claim 1;

d) reacting the intermediate of formula (II) with an appropriate formaldehyde compound of formula (VI) to form a compound of formula (I) wherein t is 1, i.e. a compound of formula (I-c), or

Wherein the variables are as defined in claim 1;

e) reacting the intermediate of formula (VII) with lithium aluminium hydride in a suitable solvent to form a compound of formula (I) wherein s is 1, i.e. a compound of formula (I-d)

Wherein the variables are as defined in claim 1.

21. A method according to claim 20 wherein W is halogen.