HK1122274A - Pyrimidine derivatives - Google Patents

Description

Pyrimidine derivatives and their use in the treatment of cancer

The present invention relates to novel compounds and processes for their preparation, methods of treating diseases, particularly cancer, comprising administering said compounds, and methods of preparing pharmaceutical compositions for treating or preventing diseases, particularly cancer.

Nitrogen-containing heterocyclic compounds such as pyrimidine derivatives, which have a variety of pharmaceutical properties and uses, have been described in patent and non-patent publications. Some publications are exemplified below.

WO 03/062225(Bayer) relates to pyrimidine derivatives as rho kinase inhibitors and their use in the treatment of rho kinase mediated diseases including cancer.

WO 2001/87845(Fujisawa) relates to nitrogen-containing heterocyclic compounds having 5-HT antagonistic activity. These compounds are known to be useful in the treatment or prevention of central nervous system disorders.

WO 95/10506(Du Pont Merck) relates to 1N-alkyl-N-aryl pyrimidinamines and derivatives thereof, which are known to inhibit Corticotropin Releasing Factor (CRF) peptides and are useful in the treatment of psychiatric and neurological disorders.

WO 2004/048365(Chiron) relates to 2, 4, 6-trisubstituted pyrimidines as inhibitors of Phosphatidylinositol (PI) 3-kinase and their use in the treatment of cancer. WO 2004/000820(Cellular Genomics) relates to nitrogen-containing heterocyclic and other compounds as kinase modulators and their use in treating a variety of kinase-associated diseases, including cancer.

WO 01/62233(Hoffmann La Roche) relates to nitrogen containing heterocyclic compounds and their use in the treatment of diseases modulated by adenosine receptors.

US 2004/0097504(Vertex) relates to nitrogen containing heterocyclic compounds useful in the treatment of various protein kinase mediated diseases.

There is a constant interest in the pharmaceutical field to identify new pharmaceutically active compounds. Such materials are the subject of the present application.

In one embodiment, the present invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof:

wherein

A represents an oxygen atom or a group-NR^AWherein R is^ARepresents H or alkyl;

d represents a-CH-unit or a nitrogen atom;

R²represents a bicyclic aromatic ring system, wherein said bicyclic aromatic ring system may be optionally substituted with 0, 1 or 2 substituents independently selected from: alkyl, trifluoromethyl, halogen, alkoxy, hydroxy, amino, dialkylamino, amido, aminocarbonyl, alkylaminocarbonyl, and dialkylaminocarbonyl; or

R²Represents a group

It may be optionally substituted with 0, 1 or 2 substituents independently selected from: alkyl, trifluoromethyl, halogen, alkoxy, hydroxy, amino, alkylamino, alkylcarbonylamino, aminocarbonyl and alkylaminocarbonyl; or

R²Represents a 1, 3-benzodioxolane which may be optionally substituted with 0, 1 or 2 substituents independently selected from: alkyl, trifluoromethyl, halogen, alkoxy, hydroxy, amino, alkylamino, alkylcarbonylamino, aminocarbonyl and alkylaminocarbonyl;

R³represents chlorine, cyano, aminocarbonyl, alkylaminocarbonyl, alkyl or trifluoromethyl,

R⁴represents H or alkyl;

R⁵represents H or halogen.

In another embodiment, the invention relates to a compound of formula (Ic) or a pharmaceutically acceptable salt thereof:

wherein

R^2-1Represents naphthyl or 1, 3-benzodioxolyl;

R^3-1represents an alkyl group, a cyano group, an aminocarbonyl group or a trifluoromethyl group.

In another embodiment, the invention relates to compounds of formula (Ic),

wherein R is^2-1Represents 1-naphthyl or 5- (1, 3-benzodioxolyl).

In another embodiment, the invention relates to a compound of the following formula (Id):

wherein

R^2-2Represents naphthyl, indolyl, furyl, benzothienyl, N-methylindolyl, 1, 3-benzodioxolyl or a radical

R^3-2Represents a methyl group, a cyano group, an aminocarbonyl group or a trifluoromethyl group.

In another embodiment, the invention relates to a compound of formula (Id), or a pharmaceutically acceptable salt thereof,

wherein

R^2-2Represents naphthyl, 5-indolyl, 2-furyl, 2-benzothienyl, 5- (N-methyl) indolyl, 5- (1, 3-benzodioxolyl) or a group

The compounds of the present invention may exist in stereoisomeric forms (enantiomers or diastereomers) depending on their structure. The present invention therefore relates to enantiomers or diastereomers and to the corresponding mixtures thereof. Mixtures of these enantiomers or diastereomers may be separated into the stereoisomeric building blocks by known methods.

Unless otherwise indicated, the following definitions apply to all technical expressions used in the present description and claims:

salt: salts for the purposes of the present invention are preferably pharmaceutically acceptable salts of the compounds of the invention.

Pharmaceutically acceptable salts: pharmaceutically acceptable salts of compound (I) include acid addition salts of inorganic acids, carboxylic acids and sulfonic acids, such as hydrochloride, hydrobromide, sulfate, phosphate, methanesulfonate, ethanesulfonate, toluenesulfonate, benzenesulfonate, naphthalenedisulfonate, acetate, propionate, lactate, tartrate, malate, citrate, fumarate, maleate and benzoate.

Pharmaceutically acceptable salts of compound (I) also include salts with common bases, such as and preferably alkali metal salts (e.g., sodium and potassium salts), alkaline earth metal salts (e.g., calcium and magnesium salts), and ammonium salts derived from ammonia or organic amines having 1 to 16 carbon atoms, such as, for example, exemplary and preferred organic amines are ethylamine, diethylamine, triethylamine, ethyldiisopropylamine, monoethanolamine, diethanolamine, triethanolamine, dicyclohexylamine, dimethylaminoethanol, procaine, dibenzylamine, N-methylmorpholine, dihydroabietylamine, arginine, lysine, ethylenediamine, and methylpiperidine.

Alkyl represents straight or branched chain alkyl groups typically having 1 to 6, 1 to 4 or 1 to 3 carbon atoms, exemplified by methyl, ethyl, n-propyl, isopropyl, tert-butyl, n-pentyl and n-hexyl.

Alkoxy represents a straight or branched chain hydrocarbon group having 1 to 6, 1 to 4 or 1 to 3 carbon atoms and linked by an oxygen atom, and exemplified by methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, pentyloxy, isopentyloxy, hexyloxy, isohexyloxy. The terms "alkoxy" and "alkyloxy" are generally used synonymously.

Alkylamino represents alkylamino having 1 or 2 alkyl substituents independently selected from methylamino, ethylamino, N-propylamino, isopropylamino, tert-butylamino, N-pentylamino, N-hexylamino, N-dimethylamino, N-diethylamino, N-ethyl-N-methylamino, N-methyl-N-propylamino, N-isopropyl-N-propylamino, N-tert-butyl-N-methylamino, N-ethyl-N-pentylamino and N-hexyl-N-methylamino.

Alkylaminocarbonyl denotes alkylaminocarbonyl having 1 or 2 alkyl substituents independently selected from the group consisting of methylaminocarbonyl, ethylaminocarbonyl, N-propylaminocarbonyl, isopropylaminocarbonyl, tert-butylaminocarbonyl, N-pentylaminocarbonyl, N-hexylaminocarbonyl, N-dimethylaminocarbonyl, N-diethylaminocarbonyl, N-ethyl-N-methylaminocarbonyl, N-methyl-N-propylaminocarbonyl, N-isopropyl-N-propylaminocarbonyl, N-tert-butyl-N-methylaminocarbonyl, N-ethyl-N-pentylamino-carbonyl and N-hexyl-N-methylaminocarbonyl are exemplary.

Aryl represents a monocyclic to tricyclic carbocyclic group in which at least one ring is aromatic and which is linked via an oxygen atom, typically having 6 to 14 carbon atoms, exemplified by phenyl, naphthyl and phenanthryl.

A bicyclic aromatic ring system means a ring system consisting of 2 fused aromatic rings, the ring system containing up to 12 ring atoms of which 3 may be heteroatoms independently selected from S, O or N.

Halogen represents fluorine, chlorine, bromine or iodine.

Immediately after the key^*The symbol refers to the point of attachment in the molecule.

For the sake of brevity, the term "without numerical limitation" when used throughout this document generally includes plural terms unless otherwise specified. For example, the term "a method of treating a disease in a subject comprising administering to the subject an effective amount of a compound of claim 1" is meant to encompass the simultaneous treatment of more than one disease and the administration of more than one compound of claim 1.

If a group in a compound of the invention is substituted, the group may be substituted with one or more substituents which may be the same or different, unless otherwise specified. Preference is given to substitution with up to 3 identical or different substituents. Very particular preference is given to substitution with 1 substituent.

In another embodiment, the invention provides a process for the preparation of a compound of formula (I) comprising contacting a suitable Pd catalyst, e.g. Pd₂(dba)₃[ Tris (dibenzylideneacetone) dipalladium (0)]、Pd(PPh₃)₄[ tetrakis (triphenylphosphine) palladium (0)]Or PdCl₂(dppf)·CH₂Cl₂{ [1, 1' -bis (diphenylphosphino) ferrocene]Palladium (II) dichloride in the presence of a dichloromethane complex } reacting a precursor of formula (II):

wherein A, D and R³-R⁵Has the meaning of the above-mentioned formula,

[A] with a reagent of formula (IIIa):

wherein R is²Having the above meaning, R¹¹And R¹²May be H or alkyl, or

[B] With a reagent of formula (IIIb):

wherein R is²Have the above-mentioned meanings.

The compounds of formula (II) can be prepared by condensing a parent of formula (IV) below with 2-amino-4, 6-dichloropyrimidine,

wherein R is³-R⁵Have the above-mentioned meanings.

The compounds of the formulae (IV), (IIIa) and (IIIb) are known or can be prepared analogously to known methods.

It is also understood that the starting materials are commercially available or can be readily prepared by standard methods well known in the art. These methods include, but are not limited to, the transformation methods listed herein.

Unless otherwise stated, the reaction is usually carried out in an inert organic solvent, which does not change under the reaction conditions. These solvents include ethers (e.g. diethyl ether, 1, 4-dioxane or tetrahydrofuran), halogenated hydrocarbons (e.g. dichloromethane, trichloromethane, carbon tetrachloride, 1, 2-dichloroethane, trichloroethane or tetrachloroethane), hydrocarbons (e.g. benzene, toluene, xylene, hexane, cyclohexane or mineral oil fractions), alcohols (e.g. methanol, ethanol or isopropanol), nitromethane, dimethylformamide or acetonitrile. Mixtures of solvents may also be used.

The reaction is generally carried out at a temperature in the range of from 0 ℃ to 150 ℃, preferably from 0 ℃ to 70 ℃. The reaction can be carried out at atmospheric pressure, elevated pressure or reduced pressure (e.g. 0.5 to 5 bar). In general, the reaction is carried out under atmospheric pressure of air or an inert gas, usually nitrogen.

The preparation of the compounds of the present invention can be illustrated by the following synthetic scheme 1:

scheme 1

Many of the compounds of the present invention exhibit useful pharmacological and pharmacokinetic properties. They are therefore useful in the treatment or prevention of diseases, especially hyperproliferative diseases such as cancer, in humans and animals.

In another embodiment, the present invention provides a pharmaceutical composition comprising at least one compound of the present invention. In another embodiment, the present invention provides a pharmaceutical composition comprising at least one compound of the present invention and one or more pharmacologically safe excipients or carrier materials. In yet another embodiment, the invention provides the use of the compounds and compositions for the treatment of diseases, and methods for treating diseases by administering to a patient a therapeutically effective amount of the compounds or compositions.

If used as active compounds, the compounds of the invention are preferably isolated in substantially pure form, that is to say substantially free of residues from the synthesis. Purity can be determined by methods known to the chemist or pharmacist (see Remington's pharmaceutical sciences, 18 th edition, 1990, Mack Publishing Group, Enolo). The purity of the compounds is preferably > 99% (w/w), but purities of > 95%, > 90% or > 85% may also be used, if desired.

The invention also relates to methods of treating or preventing a hyperproliferative disorder in a mammal, or in the manufacture of a medicament for treating or preventing a hyperproliferative disorder in a mammal, using a compound or composition described herein. The method comprises administering to a patient (or mammal), including a human, in need thereof an amount of a compound of the present invention, a pharmaceutically acceptable salt or ester thereof, or a composition of the present invention effective in treating or preventing a disease.

Hyperproliferative diseases include, but are not limited to, solid tumors such as breast cancer, respiratory tract cancer, brain cancer, genital cancer, digestive tract cancer, urinary tract cancer, eye cancer, liver cancer, skin cancer, head and neck cancer, thyroid cancer, parathyroid cancer and their distant metastases. These diseases also include lymphomas, sarcomas and leukemias.

The invention also relates to methods of preventing hyperproliferative diseases in mammals described herein using the compounds of the present invention as prophylactic or chemopreventive agents. The method comprises administering to a mammal, including a human, in need thereof an amount of a compound of the present invention or a pharmaceutically acceptable salt or ester thereof effective to delay or reduce the onset of the disease.

Examples of breast cancer include, but are not limited to, invasive ductal carcinoma, invasive lobular carcinoma, ductal carcinoma in situ, and lobular carcinoma in situ.

Examples of respiratory cancers include, but are not limited to, small cell lung cancer, non-small cell lung cancer, and bronchial adenomas and pleuropneumocytomas.

Examples of brain cancers include, but are not limited to, brain stem glioma, pituitary (hypophtalmic) glioma, cerebellar astrocytoma, cerebral astrocytoma, medulloblastoma, ependymoma, and neuroectodermal and pineal tumors.

Male genital tumors include, but are not limited to, prostate cancer and testicular cancer. Female genital tumors include, but are not limited to, endometrial, cervical, ovarian, vaginal, and vulvar cancers, as well as uterine sarcomas.

Tumors of the digestive tract include, but are not limited to, anal, colon, colorectal, esophageal, gallbladder, gastric, pancreatic, rectal, small intestine, and salivary gland cancers.

Urethral tumors include, but are not limited to, bladder, penile, kidney, renal pelvis, ureter, and urethral cancers.

Eye cancers include, but are not limited to, intraocular melanoma and retinoblastoma.

Examples of liver cancers include, but are not limited to, hepatocellular carcinoma (hepatocellular carcinoma with or without fibrolamellar variation), cholangiocarcinoma (intrahepatic cholangiocarcinoma), and mixed hepatocellular cholangiocarcinoma.

Skin cancers include, but are not limited to, squamous cell carcinoma, Kaposi's sarcoma, malignant melanoma, Merkel cell skin cancer (Merkel cell skin cancer), and non-melanoma skin cancers.

Head and neck cancers include, but are not limited to, laryngeal/laryngopharyngeal/nasopharyngeal/oropharyngeal cancer, as well as lip and oral cancers.

Lymphomas include, but are not limited to, AIDS-related lymphomas, non-Hodgkin's lymphoma, cutaneous T-cell lymphomas, Hodgkin's disease, and central nervous system lymphomas.

Sarcomas include, but are not limited to, soft tissue sarcomas, osteosarcomas, malignant fibrous histiocytomas, lymphosarcomas, and rhabdomyosarcomas.

Leukemias include, but are not limited to, acute myelogenous leukemia, acute lymphoblastic leukemia, chronic lymphocytic leukemia, chronic myelogenous leukemia, and hairy cell leukemia.

These diseases in humans are well characterized and other mammals also have diseases with similar etiologies that can be treated by administering the compounds and/or pharmaceutical compositions of the present invention.

In another embodiment, the invention provides a medicament containing at least one compound of the invention. In another embodiment, the present invention provides a medicament containing at least one compound of the invention and one or more pharmacologically safe excipients or carrier substances, such as hydroxypropyl cellulose, and the use thereof for the above-mentioned purposes.

The active ingredient may act systemically and/or locally. For this purpose, it can be applied in a suitable manner, for example orally, parenterally, pulmonarily, nasally, sublingually, lingually, buccally, rectally, transdermally, conjunctivally, otically or as an implant.

For these application routes, the active ingredient can be administered in the form of a suitable application. For a summary of the application forms, see Remington's Pharmaceutical Sciences, 18 th edition, 1990, Mack Publishing Group, Enolo.

Useful oral application forms include those which release the active ingredient rapidly and/or in modified form, for example tablets (uncoated and coated, e.g. with enteric coating), capsules, coated tablets, granules, pellets, powders, emulsions, suspensions, solutions and aerosols. Such sustained release pharmaceutical compositions are described in Remington's pharmaceutical sciences, 18 th edition, Chapter 91, part 8, 1990, Mack Publishing Group, Enolo.

Parenteral application can be carried out to avoid an absorption step (intravenous, intra-arterial, intracardiac, intraspinal or intralumbar) or to include absorption at the same time (intramuscular, subcutaneous, intradermal, transdermal or intraperitoneal). Useful parenteral application forms include injection and infusion preparations in the form of solutions, suspensions, emulsions, lyophilisates and sterile powder injections. These parenteral Pharmaceutical compositions are described in Remington's Pharmaceutical Sciences, 18 th edition, Chapter 84, part 8, 1990, Mack Publishing Group, Enolo.

In one embodiment, the invention relates to the intravenous (i.v.) use of the active compounds, such as bolus injection (i.e. single dose, e.g. per syringe), infusion over a short period of time (e.g. up to 1 hour) or infusion over a long period of time (e.g. over 1 hour). The use may be carried out by intermittent administration. The volume used may vary with the disease, and is typically 0.5-30ml or 1-20ml for bolus injections, 25-500ml or 50-250ml for short infusion times, and 50-1000ml or 100-500ml for long infusion times.

The application form must be sterile pyrogen-free. They are solvent-based or mixed aqueous and organic solvents. Examples are ethanol, polyethylene glycol (PEG)300 or 400, aqueous solutions containing cyclodextrins or emulsifiers, such as lecithin, Pluronic F68 ®, SolutolHS15 ® or Cremophor ®. Preferably an aqueous solution.

For intravenous use, solutions are typically isotonic and hydrogen rich (euhydrac), e.g., at a pH of 3-11, 6-8, or about 7.4.

Glass or plastic containers may be used as packaging for intravenous solutions, such as rubber-sealed vials. They can hold a volume of liquid of 1-1000ml or 5-50 ml. The solution may be drawn directly from the vial for use by the patient. For this purpose, it is preferred to provide the active compound in solid form (e.g., as a lyophilizate) and to dissolve it by adding the solvent to the vial immediately prior to administration.

The infusion solutions are preferably contained in containers made of glass or plastic (e.g., bottles) or collapsible containers (e.g., bags). They can hold a volume of liquid of 1-1000ml or 50-500 ml.

Forms suitable for other routes of application include, for example, inhalable pharmaceutical forms (including powder inhalants, sprays), nasal drops/drops, sprays; tablets or capsules for lingual, sublingual or buccal administration, suppositories, otic preparations, ophthalmic preparations, vaginal capsules, aqueous suspensions (lotions, concussions), lipophilic suspensions, ointments, creams, emulsions, pastes, dusting powders or implants.

The active ingredient can be converted in a manner known per se into the stated application forms. This can be done with inert, non-toxic, pharmaceutically suitable excipients. These include, inter alia, carriers (for example microcrystalline cellulose), solvents (for example liquid polyethylene glycol), emulsifiers (for example sodium lauryl sulfate), dispersants (for example polyvinylpyrrolidone), synthetic and natural biopolymers (for example self-protein), stabilizers (for example antioxidants such as ascorbic acid), colorants (for example inorganic pigments such as iron oxide) or taste and/or odor flavorings. Section C of this application gives an exemplary form of application.

For human use, in the case of oral administration, it is recommended to administer a dose of 0.001-50mg/kg, or 0.01-20 mg/kg. In the case of parenteral administration, for example intravenously or via the nasal mucosa, the oral mucosa or by inhalation, a dosage of 0.001 to 0.60mg/kg, in particular 0.01 to 30mg/kg, is recommended.

Nevertheless, in some cases, it may be necessary to deviate from the stated dosages, that is to say dosages which vary with the body weight, the route of application, the individual condition of the active ingredient, the method of preparation and the time or interval at which the application takes place. For example, in some cases it may be sufficient to use less than the aforementioned minimum amount, while in other cases the upper limit mentioned has to be exceeded. In the case of larger amounts, it is advantageous to divide them into a plurality of individual doses over the entire day.

Unless otherwise indicated, percentages in the experiments and examples are by weight; parts are parts by weight. The solvent ratio, dilution ratio and concentration of the liquid/liquid solution are given by volume.

A.Examples

Abbreviations and acronyms

A general list of abbreviations used by those of ordinary skill in the art is published in the first phase of Journal of Organic Chemistry; the directory is typically provided in tabular form, with a title name ofStandard List of Abbreviations(Standard abbreviation list). The abbreviations contained in this table and all abbreviations used by those of ordinary skill in the art of organic chemistry are incorporated herein by reference.

For the purposes of the present invention, chemical elements are identified according to the periodic Table of the chemical elements (Handbook of Chemistry and Physics, 67 th edition, 1986-87, CAS version).

More specifically, when the following abbreviations are used throughout the specification, the meanings are as follows:

2X 2 times

3X 3 times

AlMe₃Trimethylaluminum

Boc tert-butoxycarbonyl

n-BuLi butyl lithium

t-BuOK Potassium tert-butoxide

calcd calculated value

Celite ® diatomaceous earth filter, a registered trademark of Celite corporation

CD₃OD methanol-d₄

CHCl₃-d chloroform-d

d double peak

DBU 1, 8-diazabicyclo [5.4.0] undec-7-ene

DCC dicyclohexylcarbodiimide

DEAD azodicarboxylic acid diethyl ester

DIBAH diisobutylaluminum hydride

DIEA diisopropylethylamine

DMA dimethyl acetamide

DMAP 4-dimethylaminopyridine

DME dimethoxyethane

DMF N, N-dimethylformamide

DMSO dimethyl sulfoxide

DMSO-d₆Dimethyl sulfoxide-d₆

EDCI 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride

EtSH ethanethiol

EtOAc ethyl acetate

EtOH ethanol

Et₃SiH triethylsilane

h hours

HATU hexafluorophosphate O- (7-azabenzotriazol-1-yl) -N, N, N', N-

Tetramethyluronium

Hex Hexane

¹HNMR proton NMR

HOAc acetic acid

HPLC high performance liquid chromatography

LC-MS liquid chromatography/Mass Spectrometry

LDA lithium diisopropylamide

LiHMDS lithium hexamethyldisilazide

m multiplet

m-CPBA 3-chloroperoxybenzoic acid

MeOH methanol

min for

Me₃SiI trimethyl iodosilane

MS ES electrospray mass spectrometry

NaBH(OAc)₃Sodium triacetoxyborohydride

OMs O-methylsulfonyl

OTs O-p-toluenesulfonyl

OTf O-trifluoroacetyl group

Pd/C palladium on carbon

Pd₂(dba)₃Tris (dibenzylideneacetone) dipalladium (0)

Pd(PPh₃)₄Tetrakis (triphenylphosphine) palladium (0)

PdCl₂(dppf)·CH₂Cl₂[1, 1' -bis (diphenylphosphino) ferrocene]Palladium (II) dichloride and dichloro

Methane complexes

Retention time of RT

rt Room temperature

R_fTLC retention factor

s single peak

t triplet peak

TFA trifluoroacetic acid

THF tetrahydrofuran

TLC thin layer chromatography

General analytical method

The structures of representative compounds of the invention were determined by the following methods.

Electron impact mass spectrometry (EI-MS) was obtained using a Hewlett Packard5989A mass spectrometer equipped with a Hewlett Packard 5890 gas chromatograph, using a J & W DB-5 column (0.25uM coating; 30m × 0.25 mm). The ion source was maintained at 250 ℃ and spectral scans were performed at 50-800amu intervals for 2 seconds per scan.

High pressure liquid chromatography-electrospray mass spectrometry (LC-MS) was obtained as follows:

(A) Hewlett-Packard 1100 HPLC equipped with a four-stage pump, variable wavelength detector set to 254nm, YMC pro C-18 column (2X 23mm, 120A), Finnigan LCQ ion trap mass spectrometer with electrospray ionization. Spectral scanning was performed at 120-1200amu intervals with variable ion time (variable ion time) depending on the ion number of the ion source. Eluent A: 2% aqueous acetonitrile + 0.02% TFA, B: 2% aqueous acetonitrile + 0.018% TFA. Elution was performed with a 10% -95% B gradient for 3.5 minutes at a flow rate of 1.0 ml/min for an initial 0.5 minute hold and a final 0.5 minute hold in 95% B. The total operating time was 6.5 minutes.

Or

(B) A Gilson HPLC system equipped with 2 Gilson 306 pumps, a Gilson 215 autosampler, a Gilson diode array detector, a YMC Pro C-18 column (2X 23mm, 120A), and a Z-spray electrospray ionization Micromass LCZ single quadrupole mass spectrometer. The spectral scan was performed for 1.5 seconds at the interval of 120-800 amu. ELSD (Evaporative Light Scattering Detector) data was also obtained as an analog channel. The eluent is A: 2% aqueous acetonitrile + 0.02% TFA, or B: 2% aqueous acetonitrile + 0.018% TFA. Elution was carried out with a 10% -95% B gradient for 3.5 minutes at a flow rate of 1.5 ml/min, initially for 0.5 minutes and finally for 0.5 minutes in 90% B. The total operating time was 4.8 minutes. Another switching valve is used for column switching and regeneration.

Conventional one-way NMR spectroscopy was performed using a 400 MHz Varian Mercury-plus spectrometer. The sample was dissolved in deuterated solvent from Cambridge Isotope Labs and transferred to a 5mm ID Wilmad NMR tube. Spectra were obtained at 293K. Recording chemical shifts in the ppm range with reference to appropriate solvent signals, e.g.¹For DMSO-d in H spectrum₆2.49ppm or for CD₃CN-d₃1.93ppm for CD₃OD 3.30ppm for CD₂Cl₂-d₂5.32ppm for CHCl₃D is 7.26 ppm.

General HPLC purification method

Preparative reverse phase HPLC chromatography was carried out using a Gilson 215 system, typically using a column of YMCPro-C18 AS-342 (150X 20mm I.D.). Typically, the mobile phase used is a mixture of (A) aqueous 0.1% TFA and (B) acetonitrile. The usual gradient is:

time [ min ]]	A：％	B：％	Flow rate [ ml/min ]]
				0.50	90.0	10.0	1.0
11.00	0.0	100.0	1.0
				14.00	0.0	100.0	1.0
15.02	100.0	0.0	1.0

Intermediate 1A: 4- {4- [ (2-amino-6-chloropyrimidin-4-yl) amino]Phenoxy } pyridine-2-carbonitriles

Step 1: preparation of 4- (4-aminophenoxy) pyridine-2-carbonitrile

4-aminophenol (41.35g, 0.38mol) and N, N-dimethylacetamide (500ml) were charged to a 3L three-necked round bottom flask equipped with a mechanical stirrer and a reflux condenser. After degassing the resulting solution by introducing nitrogen, potassium tert-butoxide (44.54g, 0.40mol) was added in portions. The solution initially turned green and then turned an off-white suspension to which was added a solution of 4-chloropyridine-2-carbonitrile (50.00g, 0.36mol) in N, N-dimethylacetamide (300ml) in one portion. The mixture turned brown within a few minutes and was heated to 90 ℃ overnight. The next morning, the mixture was cooled to room temperature and the solvent was removed in vacuo. The resulting residue was partitioned between water (1.5L) and EtOAc (1.5L). Adding K₂CO₃The pH was adjusted to slightly basic and the layers were separated. The aqueous layer was extracted with EtOAc (1L). The combined organic phases were over MgSO₄Drying, filtering and concentrating. The resulting residue was dissolved in dichloromethane and taken up with a plug of silica gel (. about.1 kg). Then eluted with 25% -75% EtOAc/hexanes to give 4- (4-aminophenoxy) pyridine-2-carbonitrile (18.9g, 25%):¹H NMR(DMSO-d₆) δ ppm 8.48(d, 1H), 7.51(d, 1H), 7.04(dd, 1H), 6.83(dd, 2H), 6.60(dd, 2H), 5.18(s, 2H); MS ES 212(M + H), retention time 0.97 min.

Step 2: preparation of the title Compound

4- (4-Aminophenoxy) pyridine-2-carbonitrile (70.00g, 0.33mol), 4, 6-dichloropyrimidin-2-amine (54.35g, 0.33mol), water (2.5L), and 2-propanol (500ml) were charged into a 3L three-necked round bottom flask equipped with a mechanical stirrer and a reflux condenser. After heating the suspension to 91 ℃ for 4 hours, it was allowed to cool to room temperature overnight. The reaction mixture was filtered and the collected solid was washed with EtOH, ether and hexane. The solid was dried by suction for 45 minutes to give 4- {4- [ (2-amino-6-chloropyrimidin-4-yl) amino group]Phenoxy } pyridine-2-carbonitrile (84.1g, 75%):¹H NMR(DMSO-d₆)δ ppm 9.45(s，1H)，8.55(d，1H)，7.80(d，2H)，7.64(d，1H)，7.12-7.15(m，3H)，6.76(s，2H)，6.00(s，1H)，3.34(s，2H) (ii) a MS ES 339(M + H), retention time 2.49 min.

Intermediate 1B: 6-chloro-N⁴- (4- { [2- (trifluoromethyl) pyridin-4-yl]Oxy } phenyl) pyrimidine-2, 4-diamines

Prepared in a two-step sequence analogous to intermediate 1A:¹H NMR(DMSO-d₆) Δ ppm9.46(s, 1H), 8.59(d, 1H), 7.81(d, 2H), 7.37(d, 1H), 7.17(d, 2H), 7.11(dd, 1H), 6.78(s, 2H), 6.00(s, 1H). MS ES 382(M + H), calculated 382, retention time 2.93 min.

Intermediate 1C: 6-chloro-N⁴- {4- [ (2-methylpyridin-4-yl) oxy]Phenyl } pyrimidine-2, 4-diamines

Prepared in a two-step sequence analogous to intermediate 1A:¹H NMR(DMSO-d₆)δ 9.40(s，1H)，8.27(d，1H)，7.76(d，2H)，7.06(d，2H)，6.75(brs，2H)，6.72(d，1H)，6.66(s，1H)，5.98(s，1H)；MS ES 328(M+H)⁺calculated value 328, retention time 1.45 minutes.

Intermediate 1D: 4- {3- [ (2-amino-6-chloropyrimidin-4-yl) amino]Phenoxy } pyridine-2-carbonitriles

Prepared in a two-step sequence analogous to intermediate 1A:¹H NMR(DMSO-d₆)δ 9.52(s，1H)，8.57(d，1H)，7.72(dd，1H)，7.69(d，1H)，7.53(dd，1H)，7.38(dd，1H)，7.18(dd，1H)，6.77-6.80(m，3H)，6.01(s，1H)；MS ES 339(M+H)⁺calculated 339, retention time 2.65 minutes.

Intermediate 2A: 4- {3- [ (2-amino-6-chloropyrimidin-4-yl) amino]Phenoxy } pyridine-2-carboxamides

To a 100ml round bottom flask was added 4- {3- [ (2-amino-6-chloropyrimidin-4-yl) amino]Phenoxy } pyridine-2-carbonitrile (intermediate 1D, 5.00g, 14.8mmol) and concentrated sulfuric acid (40 ml). After heating the mixture to 70 ℃ for 2 hours, it was allowed to cool to room temperature. Then NaHCO was slowly poured in₃And ice water, and EtOAc was added with stirring. The organic layer was separated over MgSO₄Drying and filtering. The filtrate was concentrated in vacuo to give 4- {3- [ (2-amino-6-chloropyrimidin-4-yl) amino group]Phenoxy } pyridine-2-carboxamide (4.50g, 85%, colorless powder): δ 9.51(s, 1H), 8.49(d, 1H), 8.13(d, 1H), 7.72(d, 1H), 7.66-7.68(m, 1H), 7.54(dd, 1H), 7.43(d, 1H), 7.38(dd, 1H), 7.18(dd, 1H), 6.77-6.81(m, 3H), 6.00(s, 1H); MS ES357(M + H)⁺Calculated 357, retention time 2.32 minutes.

Example 1 and example 2: High-Speed analog (HSA) synthesis Method B

1 equivalent of 4- {4- [ (2-amino-6-chloropyrimidin-4-yl) amino group in a 5ml microwave reaction vessel]Phenoxy } pyridine-2-carbonitrile (100mg, intermediate 1A), 2 equivalents of 1-naphthylboronic acid and 0.06 equivalents of PdCl₂(dppf)·CH₂Cl₂And 2.3ml of anhydrous N, N-dimethylacetamide was added 3.1 equivalents of 2M K₂CO₃An aqueous solution. After degassing the resulting mixture with nitrogen for 10 minutes, the vial was sealed and placed in a microwave reactor (Emrys o)ptimizer, manufactured by Personal Chemistry) at 150 ℃ for 20 minutes. The reaction mixture was filtered, the filtrate was concentrated and purified by preparative HPLC (using Phenomenex Luna 5 μ C18150 × 30mm column, elution 15% -85% acetonitrile) to give the final product. Example 2 is a byproduct of the hydrolysis in the reaction of example 1.

Example 14 was prepared as described in example 1 using the appropriate starting materials.

Example 3: high Speed Analog (HSA) synthesis method C

1 equivalent of 4- {3- [ (2-amino-6-chloropyrimidin-4-yl) amino group in an 8ml microwave reaction vessel]Phenoxy } pyridine-2-carboxamide (100mg, intermediate 2A), 2 equivalents of 1, 3-benzodioxol-5-ylboronic acid and 0.06 equivalents of PdCl₂(dppf)·CH₂Cl₂To a mixture of the complex and 2.3ml of anhydrous N, N-dimethylacetamide was added 3.1 equivalents of 2M K₂CO₃An aqueous solution. After the resulting mixture was degassed with nitrogen for 10 minutes, the vial was sealed and heated in a microwave reactor (Emryscopizer, manufactured by Personal Chemistry Co.) at 140 ℃ for 20 minutes. The reaction mixture was filtered, the filtrate was concentrated and purified by preparative HPLC (using phenomenex luna 5 μ C18150 × 30mm column, elution 15% -85% acetonitrile) to give the final product.

Examples 4-9 were prepared as described in example 3 using the appropriate starting materials.

Example 10: high Speed Analog (HSA) Synthesis method A

In a microwave reactor (Emrys optizer, manufactured by Personal chemistry Co., Ltd.) under nitrogen atmosphere, 1 equivalent of 6-chloro-N in 5ml of a microwave reaction vessel⁴- (4- { [2- (trifluoromethyl) pyridin-4-yl]Oxy } phenyl) pyrimidine-2, 4-diamine (100mg, intermediate 1B), 2 equivalents of 1, 3-benzodioxol-5-ylboronic acid and 0.1 equivalent of PdCl₂(dpPf)-CH₂Cl₂Mixture of Complex and 2.5ml of anhydrous N, N-dimethylacetamide and 0.5ml of 2MK₂CO₃Dissolving in waterThe solution was heated at 140 ℃ for 20 minutes. The reaction mixture was filtered, the filtrate was concentrated and purified by preparative HPLC (using Phenomenex Luna 5 μ C18150 × 30mm column eluting with 15% -85% acetonitrile containing 0.1% TFA) to give the final product.

Example 11 was prepared as described for example 10 using the appropriate starting materials.

Example 12: 6- (1H-indol-5-yl) -N⁴- (4- { [2- (trifluoromethyl) pyridin-4-yl]Preparation of oxy } phenyl) pyrimidine-2, 4-diamines

To 1 equivalent of 6-chloro-N in a round bottom flask⁴- (4- { [2- (trifluoromethyl) pyridin-4-yl]Oxy } phenyl) pyrimidine-2, 4-diamine (200mg, intermediate 1B), 2 equivalents of 1H-indol-5-yl boronic acid and 0.06 equivalents of PdCl₂(dppf) and 4ml of anhydrous N, N-dimethylacetamide was added to a mixture of 3.1 equivalents of 2M K₂CO₃An aqueous solution. After the resulting mixture was degassed with nitrogen for 10 minutes, it was heated at 120 ℃ overnight under nitrogen blanket. The reaction mixture was cooled and filtered, the filtrate was concentrated and purified by HPLC (mixture of water and acetonitrile, gradient from initial acetonitrile 15% to final acetonitrile 85%) to give 6- (1H-indol-5-yl) -N⁴- (4- { [2- (trifluoromethyl) pyridin-4-yl]Oxy } phenyl) pyrimidine-2, 4-diamine:¹H NMR(DMSO-d₆)δ11.22(s，1H)，9.31(s，1H)，8.60(m，1H)，8.19(s，1H)，7.90(m，2H)，7.69(m，1H)，7.43(d，1H)，7.38(m，2H)，7.18-7.10(m，3H)，6.52(m，2H)，6.30(s，2H)；MS ES463(M+H)⁺calculate a value 463.

Example 13 was prepared as described for example 12 using the appropriate starting materials.

Example 15: 6- [3- (morpholin-4-ylmethyl) -1H-indol-5-yl]-N⁴- (4- { [2- (trifluoromethyl) pyridin-4-yl]Oxy } phenyl) pyrimidine-2, 4-bisPreparation of amines

To a 20ml round bottom flask was added 6- (1H-indol-5-yl) -N⁴- (4- { [2- (trifluoromethyl) pyridin-4-yl]Oxy } phenyl) pyrimidine-2, 4-diamine (100mg, 0.22mmol), dichloromethane (4ml), acetic acid (1ml), morpholine (19mg, 0.22mmol) and formaldehyde (37% in water, 0.02 ml). The reaction mixture was stirred for 5 hours, then diluted with dichloromethane and basified with 2M NaOH. After adding EtAOc to dissolve all precipitated material, water was added and the layers were separated. The aqueous layer was extracted with EtOAc and the combined organic layers were washed with brine, over Na₂SO₄Drying, filtering and concentrating. The resulting residue was purified by preparative HPLC (with 5-75% acetonitrile/water/0.1% TFA) to afford material containing a small amount of impurities, which was repurified by preparative HPLC (with 30-45% acetonitrile/water/0.1% TFA). Purification by preparative TLC (10% MeOH/dichloromethane) finally gave 7.6mg (6.3%) of pure product:¹H NMR(CD₂Cl₂)δ 8.44(m，1H)，8.26(m，2H)，7.68(m，1H)，7.53(d，2H)，7.3 1(d，1H)，7.1 5(d，1H)，7.08(s，1H)，7.01(d，2H)，6.93(m，1H)，6.63(s，1H)，6.48(s，1H)，5.23(m，1H)，4.90(s，2H)，3.63(s，2H)，3.56(m，4H)，2.40(m，4H)；MS ES562(M+H)⁺calculated 562, retention time 2.26 minutes.

The list of examples is as follows:

B. physiological activity

The utility of the compounds of the invention can be illustrated, for example, by the in vitro activity of the compounds in the in vitro tumor cell proliferation assay described below. The relationship between the activity of in vitro tumor cell proliferation assays and clinical antitumor activity is well understood in the art. For example, in vitro tumor proliferation experiments have demonstrated the therapeutic efficacy of the following drugs: taxol (Silvestrini et al, Stem Cells 1993, 11(6), 528-35), taxotere (Bissery et al, Anti cancer rugs 1995, 6(3), 339) and topoisomerase inhibitors (Edelman et al, cancer Chemothers. Pharmacol.1996, 37(5), 385-93).

The following experiments may demonstrate the in vitro efficacy of the compounds of the invention:

cytotoxic Activity of Compounds of the invention

The following section describes assays that may be used to characterize the compounds of the present invention, e.g., to test the cytotoxic activity of the compounds on cells.

Human tumor cells such as HCT116 cells are treated at 3.0X 10³Individual cells/well were seeded into 96-well plates and incubated at 37 ℃ with 5% CO₂In an incubator, 100. mu.l of RPMI complete medium (Invitrogen corporation, Grand Island, NY) containing 10% fetal bovine serum (Hyclone, Logan, Utah) and 10mM HEPES was grown for 16 hours. To each well was added 50. mu.l of growth medium containing compounds at a concentration of 20. mu.M-60 nM in 0.2% DMSO. Cells were allowed to grow for a further 72 hours at 37 ℃. Mu.l of Alamar Blue (Trek Diagnostic Systems, Inc., Cleveland, Ohio) reagent was added to each well and incubated at 37 ℃ for 4 hours. Plates were read with SpectraMax Gemini (Molecular Devices, CA) at 544nm excitation wavelength and 590nm emission wavelength. Determination of IC by Linear regression analysis of logarithm of drug concentration versus percent inhibition₅₀The value is obtained.

The cytotoxicity of representative compounds of the invention was tested using the experimental method described above, with the following results:

in the HCT116 cytotoxic activity assay, examples 1, 2, 5, 8, 9, 10, 11, 12, 13 and 14 show results that are IC₅₀≤500nM。

In the HCT116 cytotoxic activity assay, examples 3, 4, 6, 7 and 15 show results of 500nM < IC₅₀≤5μM。

C. Related operative examples of pharmaceutical compositions

The compounds of the invention can be converted into the following pharmaceutical preparations:

tablet formulation：

Composition of：

100mg of the compound according to example 1, 50mg of lactose (monohydrate), 50mg of corn starch (local product), 10mg of polyvinylpyrrolidone (PVP 25) (from BASF, Ludwigshafen, Germany) and 2mg of magnesium stearate.

The tablet had a weight of 212mg, a diameter of 8mm and a radius of curvature of 12 mm.

Preparation of：

The mixture of active ingredient, lactose and starch was granulated together with 5% (w/w) aqueous PVP solution. After drying, the granules were mixed with magnesium stearate for 5 minutes. The mixture is molded using a conventional tablet press (tablet form, see above). The applied molding pressure is typically 15 kN.

Suspension for oral administration：

Composition of：

1000mg of the compound of example 1, 1000mg of ethanol (96%), 400mg of Rhodigel (xanthan gum from FMC, Pennsylvania, USA) and 99g of water.

A10 ml oral suspension provides a single dose of 100mg of a compound of the invention.

Preparation of：

The Rhodigel was suspended in ethanol and the active ingredient was added to the suspension. Water was added with stirring. Stirring was continued for about 6 hours until the Rhodigel had fully swelled.

Solution for intravenous administration 1：

Composition of：

100-200mg of the compound of example 1, 15g of polyethylene glycol 400 and 250g of water. In saline, optionally with up to 15% of Cremophor EL, optionally up to 15% of ethanol and optionally up to 2 equivalents of a pharmaceutically suitable acid (e.g. citric acid or hydrochloric acid).

Preparation of：

The compound of example 1 and polyethylene glycol 400 were dissolved in water with stirring. The solution was sterilized by filtration (pore size 0.22 μm) and filled into heat sterilized infusion bottles under sterile conditions. The infusion bottle was sealed with a rubber seal.

Solution for intravenous administration 2

Composition of：

100-200mg of the compound of example 1, a saline solution, optionally up to 15% by weight of polyoxyethylated castor oil, optionally up to 15% by weight of ethanol and optionally up to 2 equivalents of a pharmaceutically suitable acid, for example citric acid or hydrochloric acid.

Preparation of：

The compound of example 1 was dissolved in the aqueous salt solution with stirring. Optionally, a polyoxyethylated castor oil, ethanol or acid is added. The solution was sterilized by filtration (pore size 0.22 μm) and filled into heat sterilized infusion bottles under sterile conditions. The infusion bottle was sealed with a rubber seal.

Other embodiments of the invention will be apparent to those skilled in the art from consideration of the specification or practice of the invention disclosed herein. It is intended that the specification and examples be considered as exemplary only, with a true scope and spirit of the invention being indicated by the following claims.

Claims (15)

1. A compound of formula (I) or a pharmaceutically acceptable salt thereof:

wherein

A represents an oxygen atom or a group-NR^AWherein R is^ARepresents H or alkyl;

d represents a-CH-unit or a nitrogen atom;

R²represents a bicyclic aromatic ring system, wherein said bicyclic aromatic ring system may be optionally substituted with 0, 1 or 2 substituents independently selected from: alkyl, trifluoromethyl, halogen, alkoxy, hydroxy, amino, dialkylamino, amido, aminocarbonyl, alkylaminocarbonyl, and dialkylaminocarbonyl; or

R²Represents a group

It may be optionally substituted with 0, 1 or 2 substituents independently selected from: alkyl, trifluoromethyl, halogen, alkoxy, hydroxy, amino, alkylamino, alkylcarbonylamino, aminocarbonyl and alkylaminocarbonyl; or

R²Represents a 1, 3-benzodioxolane which may be optionally substituted with 0, 1 or 2 substituents independently selected from: alkyl, trifluoromethyl, halogen, alkoxy, hydroxy, amino, alkylamino, alkylcarbonylamino, aminocarbonyl and alkylaminocarbonyl;

R³represents chlorine, cyano, aminocarbonyl, alkylaminocarbonyl, alkyl or trifluoromethyl,

R⁴represents H or alkyl;

R⁵represents H or halogen.

2. A compound of formula (Ic) or a pharmaceutically acceptable salt thereof:

wherein

R^2-1Represents naphthyl or 1, 3-benzodioxolyl;

R^3-1represents an alkyl group, a cyano group, an aminocarbonyl group or a trifluoromethyl group.

3. The compound of claim 2 or a pharmaceutically acceptable salt thereof,

wherein R is^2-1Represents 1-naphthyl or 5- (1, 3-benzodioxolyl).

4. A process for the preparation of a compound of claim 1, which process comprises reacting a precursor of the following formula (II):

wherein A, D and R³-R⁵Having the meaning as defined in claim 1,

[A] with a reagent of formula (IIIa):

wherein R is²Has the meaning defined in claim 1, R¹¹And R¹²It may be a hydrogen atom or an alkyl group,

or

[B] With a reagent of formula (IIIb):

wherein R is²Have the meaning defined in claim 1.

5. A compound according to claim 1 for use in the treatment or prevention of a disease.

6. A pharmaceutical composition comprising a compound of claim 1.

7. The pharmaceutical composition of claim 6, further comprising at least one pharmaceutically acceptable carrier or excipient.

8. The pharmaceutical composition of claim 6 in a form suitable for oral or intravenous administration.

9. The pharmaceutical composition of claim 6 for use in the treatment or prevention of a disease.

10. A process for preparing a pharmaceutical composition according to claim 7, which process comprises mixing at least one compound according to claim 1 with at least one pharmaceutically acceptable carrier or excipient and bringing the resulting mixture into a form suitable for said pharmaceutical composition.

11. Use of a compound of claim 1 for the preparation of a pharmaceutical composition for the treatment or prevention of a disease.

12. The use of claim 11, wherein the disease is cancer.

13. A method of treating a disease or condition in a mammal, comprising administering to a mammal in need thereof an effective amount of a compound of claim 1.

14. The method of claim 13, wherein the disease or disorder is cancer.

15. A packaged pharmaceutical composition comprising a container containing the pharmaceutical composition of claim 7 and instructions for use in the treatment of a disease or condition in a mammal.