ES2366112T3 - NITROGEN HETEROCYCLIC COMPOUNDS AND PROCESS TO DO THEM. - Google Patents

Abstract

A process for making a compound according to formula A (2): Formula A (2) wherein R 1 is a member selected from the group consisting of: -CN, -O-C 1-8 alkyl having straight or branched chain ; -O-phenyl, -O-naphthyl, -O-indolyl and -O-isoquinolyl; n is from 2 to 5; R 3 is a member selected from the group consisting of: piperidine, pyrrolidine, morpholine, piperazine, 4-methyl-piperidine and 2-methyl-piperidine; and all pharmaceutically acceptable salts, hydrates and solvates thereof; comprising the steps of: (a) etherifying the hydroxy group of a compound of formula I with a compound of formula II wherein each of L1 and L2 is independently a leaving group selected from the group consisting of halogen, alkoxy, aryloxy, alkylthio, alkylsulfinyl, alkylsulfonyl, alkylsulfonyloxy and arylsulfonyloxy, wherein L1 is less reactive than the leaving group L2 under basic etherification conditions in the presence of a solvent at reflux temperature of the solvent for 2-6 hours to produce a compound of formula III as follow: ; (b) nitrate the compound according to formula III, to give a compound according to formula IV, at a temperature from 0 ° C to 80 ° C in nitric acid and a solvent, as follows:; (c) reacting the compound of formula IV, with an amine-containing compound, in the presence of a basic catalyst and a solvent to provide a compound of formula V, as follows:; (d) reduce the nitro group in the compound of formula V to an amino group and thereby produce a compound of formula VI, as follows:; (e) reacting the compound of formula VI with ammonium and formamide format at a temperature from 100 ° C to 200 ° C to produce a cyclic quinazoline derivative of formula VII, as follows:; (f) replacing the hydroxy group of the compound of formula VII with a leaving group Q selected from the group consisting of halogen, alkoxy, aryloxy, alkylthio, alkylsulfinyl, alkylsulfonyl, alkylsulfonyloxy, arylsulfonyloxy to provide a compound of formula VIII, as follows:; (g) reacting the compound of formula VIII with a compound of formula IX to replace leaving group Q and providing a compound of formula A (2), as follows: (h) and optionally, producing a salt of the compound of formula A (2).

Description

Field of the Invention

The present invention relates to nitrogen-containing heterocyclic compounds and pharmaceutically acceptable salts or prodrugs thereof that have inhibitory activity on phosphorylation of kinases, which inhibits the activity of such kinases. The invention also relates to a process for making nitrogen-containing heterocyclic compounds and intermediates thereof, and pharmaceutically acceptable salts or prodrugs thereof. The invention further relates to a method of using the compounds of the present invention by inhibiting kinases and treating disease states in a mammal by inhibiting phosphorylation of kinases by administering an effective amount of a compound according to the invention to a patient. in need of it.

Background of the invention

It is known that platelet-derived growth factor (PDGF) acts as an aggravating factor for proliferative cell diseases such as arteriosclerosis, vascular reobstruction after percutaneous coronary angioplasty and surgical revascularization, cancer, glomerulonephritis, glomerulosclerosis, psoriasis and joint rheumatism. See, Cell, 46: 155-169 (1986); Science, 253: 1129-1132 (1991); Nippon Rinsho (Japanese J. of Clinical Medicine), 50: 3038-3045 (1992); Nephrol Dial Transplant, 10: 787-795 (1995); Kidney International, 43 (Suppl. 39): 86-89 (1993); Journal of Rheumatology, 21: 1507-1511 (1994); Scandinavian Journal of Immunology, 27: 285-294 (1988).

Of the quinazoline derivatives that are useful as drugs, N, N-dimethyl-4- (6,7-dimethoxy-4quinazolinyl) -1-piperazine carboxamide is described as a bronchodilator in South African Patent No. 67 06512 (1968). The dimethoxyquinazoline derivatives are described as inhibitors of epidermal growth factor receptor (EGF) phosphorylation in published Japanese Unexamined Patent Application No. 208911/93 and WO 96/09294. Quinoline derivatives that have benzodiazepine receptor agonist activity are described in Pharmacology Biochemistry and Behavior, 53, 87-97 (1996) and European Journal of Medicinal Chemistry, 31, 417-425 (1996), and quinoline derivatives that They are useful as antiparasitic agents are described in Indian Journal of Chemistry, 26B: 550-555 (1987).

PDGF receptor phosphorylation inhibitors known so far include bismonyl and bicyclic aryl compounds and heteroaryl compounds (WO 92/20642), quinoxaline derivatives. See Cancer Research, 54: 6106 (1994), pyrimidine derivatives (published Japanese unexamined patent application No. 87834/94) and dimethoxyquinoline derivatives Summaries of the 16th Annual Meeting of the Pharmaceutical Society of Japan (Kanazawa) (1996), 2 : 275; 29 (C2): 15-21. Nitrogenous heterocyclic compounds are also described in WO 98/14431 published April 9, 1998. WO document describes various processes for making such compounds and the phosphorylation inhibition activity thereof.

Compendium of the invention

The present invention is directed to nitrogen-containing heterocyclic compounds and pharmaceutically acceptable salts thereof. These compounds have inhibitory activity on phosphorylation of kinases, which inhibits the activity of kinases. More particularly, the important inhibition of kinases according to the present invention is of receptor tyrosine kinases including platelet-derived growth factor receptor (PDGF), Flt3, CSF-1R, epidermal growth factor receptor (EGRF), factor of fibroblast growth (FGF), vascular endothelial growth factor receptor (VEGFR) and others. Another class of kinase inhibition according to the invention is non-receptor tyrosine kinase inhibitory activity including src and abl, and the like. A third class of kinase inhibition according to the invention is inhibitory activity towards serine / threonine kinases, including such kinases such as MAPK, MEK and cyclin dependent kinases (CDK) that mediates cell proliferation, AKT and CDK so that mediates cell survival and NIK that regulates inflammatory responses. The inhibition of such kinases can be used to treat diseases that involve cell survival, proliferation and migration, including cardiovascular diseases, such as arteriosclerosis and vascular resorption, cancer, glomerulosclerosis, fibrotic diseases and inflammation, as well as the general treatment of proliferative cell diseases. .

One aspect of the present invention relates to a process for making a compound according to formula A (2):

image 1

Formula A (2)

where

5 R1 is a member selected from the group consisting of: -CN, -O-C1-8 alkyl which is straight or branched chain; -O-phenyl, -O-naphthyl, -O-indolyl and -O-isoquinolyl; n is from 2 to 5; R3 is a member selected from the group consisting of: piperidine, pyrrolidine, morpholine, piperazine, 4-methyl-piperidine and 2-methyl-piperidine;

10 and all pharmaceutically acceptable salts, hydrates and solvates thereof;

which includes the steps of:

(a) etherifying the hydroxy group of a compound of formula I with a compound of formula II wherein L1 and L2 is a

15 leaving group selected from the group consisting of halogen, alkoxy, aryloxy, alkylthio, alkylsulfinyl, alkylsulfonyl, alkylsulfonyloxy, arylsulfonyloxy, wherein L1 is less reactive than the leaving group L2 under basic etherification conditions in the presence of a solvent at reflux temperature of the solvent for 2-6 hours to

image2

20 (b) nitrate the compound according to formula III, to give a compound according to formula IV, at a temperature from 0 ° C to 80 ° C in nitric acid and a solvent, as follows:

(c) reacting the compound of formula IV, with an amine-containing compound, in the presence of a basic catalyst and a solvent to provide a compound of formula V, as follows:

of formula VI, as follows:

30 100 ° C to 200 ° C to produce a cyclic quinazoline derivative of formula VII, as follows:

image3

image4

image5

image3

(F)

replace the hydroxy group of the compound of formula VII with a leaving group Q selected from the group consisting of halogen, alkoxy, aryloxy, alkylthio, alkylsulfinyl, alkylsulfonyl, alkylsulfonyloxy, arylsulfonyloxy to provide a compound of formula VIII, as follows:

(g)

 react the compound of formula VIII with a compound of formula IX to replace leaving group Q and provide a compound of formula A (2), as follows:

(h)

 and optionally, producing a salt of the compound of formula A (2).

image6

image 1

Preferably the leaving group L1 in step (a) is selected from the group consisting of Cl, Br, p-toluenesulfonate and methylsulfonate.

More preferably, the leaving group L2 in step (a) is selected from the group consisting of Cl, Br, p15 toluenesulfonate and methylsulfonate.

Advantageously, the solvent in step (a) is selected from the group consisting of toluene, methanol, ethanol, ether and THF.

Preferably, the base in step (a) is selected from the group consisting of potassium carbonate, sodium carbonate and sodium hydroxide.

Preferably further, the catalyst in step (c) is selected from the group consisting of potassium carbonate, sodium carbonate, sodium hydroxide and sodium iodide.

Advantageously, the solvent in step (c) is selected from the group consisting of toluene, ethanol, ether, THF, glyme (1,2-dimethoxyethane), diglyme and MTBE.

Preferably, the leaving group Q is selected from the group consisting of Cl, Br, p-toluenesulfonate and methylsulfonate.

Preferably further, R1 is selected from the group consisting of CN and -O-isopropyl, n is 2 or 3 and all pharmaceutically acceptable salts, hydrates and solvates thereof.

Advantageously, wherein R3 is a piperidinyl, 4-methyl-piperidinyl, 2-methyl-piperidinyl or pyrrolidinyl radical, and all pharmaceutically acceptable salts, hydrates and solvates thereof.

Preferably, where n is 3, R1 is -O-isopropyl or CN and all pharmaceutically acceptable salts, hydrates and solvates thereof.

Preferably further, wherein R3 is N-piperidine or N-pyrrolidine, and all pharmaceutically acceptable salts, hydrates and solvates thereof.

Other aspects, objects, features and advantages of the present invention will be apparent to the person skilled in the art from the following detailed description illustrating the preferred embodiments of the invention.

Detailed description of the invention

Definitions

According to the present invention and how the present document is used, the following terms are defined with the following meanings, unless explicitly specified otherwise.

The term "alkenyl" refers to a straight chain or branched chain trivalent unsaturated aliphatic radical. The term "alkynyl" refers to a straight or branched chain aliphatic radical that includes at least two carbons linked by a triple bond. If the number of alkenyl and alkynyl carbons is not specified, each refers to radicals having 2-12 carbon atoms.

The term "alkyl" refers to saturated aliphatic groups including straight chain, branched chain and cyclic groups that have the specified number of carbon atoms, or if no number is specified, that have up to 12 carbon atoms. The term "cycloalkyl" as used herein refers to a mono-, bi-or tricyclic aliphatic ring having 3 to 14 carbon atoms and preferably 3 to 7 carbon atoms.

As the terms "carbocyclic ring structure" and "mono, bi or tricyclic C3-16 carbocyclic ring structure" or the like are used herein, each is intended to mean stable ring structures that have only carbon atoms as ring atoms. wherein the ring structure is a substituted or unsubstituted member selected from the group consisting of: a stable monocyclic ring that is an aromatic ring ("aryl") having six ring atoms; a stable non-aromatic monocyclic ring having 3 to 7 ring atoms in the ring; a stable bicyclic ring structure having a total of 7 to 12 ring atoms in the two rings where the bicyclic ring structure is selected from the group consisting of ring structures in which both rings are aromatic, ring structures in the one of the rings is aromatic and ring structures in which both rings are non-aromatic; and a stable tricyclic ring structure having a total of 10 to 16 atoms in the three rings wherein the tricyclic ring structure is selected from the group consisting of: ring structures in which the three rings are aromatic, ring structures in which two of the rings are aromatic and ring structures in which the three rings are non-aromatic. In each case, the non-aromatic rings when present in the monocyclic, bicyclic or tricyclic ring structure may be independently saturated, partially saturated or completely saturated. Examples of such carbocyclic ring structures include, but are not limited to cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, adamantyl, cyclooctyl, [3.3.0] bicyclooctane, [4.3.0] biciclononano, [4.4.0] bicyclodecane (decalin), [2.2.2] bicyclooctane, fluorenyl, phenyl, naphthyl, indanyl, adamantyl or tetrahydronaphthyl (tetralin). In addition, the ring structures described herein may be attached to one.

or more indicated groups that hang from them through any carbon atom which produces a stable structure. The term "substituted" as used in conjunction with carbocyclic ring structures means that hydrogen atoms attached to the ring carbon atoms of the ring structures described herein may be substituted by one or more of the substituents indicated for that structure if such (s) substitution (s) would produce a stable compound.

The term "aryl" which is included with the term "carbocyclic ring structure" refers to a substituted or unsubstituted aromatic ring, substituted with one, two or three substituents selected from lower alkoxy, lower alkyl, lower alkylamino, hydroxy, halogen , cyano, hydroxyl, mercapto, nitro, thioalkoxy, carboxaldehyde, carboxyl, carboalkoxy and carboxamide, including, but not limited to, carbocyclic aryl, heterocyclic and baryl aryl groups and the like, all of which may be optionally substituted. Preferred aryl groups include phenyl, halophenyl, lower alkyl-phenyl, naphthyl, biphenyl, phenanthrenyl and naphthacenyl.

The term "arylalkyl" which is included with the term "carbocyclic aryl" refers to one, two or three aryl groups having the number of designated carbon atoms, attached to an alkyl group having the designated number of carbon atoms. Suitable arylalkyl groups include, but are not limited to, benzyl, picolyl, naphthylmethyl, phenethyl, benzhydryl, trityl, and the like, all of which may be optionally substituted.

As used herein, the term "heterocyclic ring" or "heterocyclic ring system" is intended to mean a substituted or unsubstituted member selected from the group consisting of stable monocyclic ring having 5-7 members in the ring itself and which it has 1 to 4 ring heteroatoms selected from the group consisting of N, O and S; a stable bicyclic ring structure having a total of 7 to 12 atoms in the two rings where at least one of the two rings has 1 to 4 heteroatoms selected from N, O and S, including bicyclic ring structures wherein any of the described stable monocyclic heterocyclic rings is fused to a hexane or benzene ring; and a stable tricyclic heterocyclic ring structure having a total of 10 to 16 atoms in the three rings where at least one of the three rings has 1 to 4 heteroatoms selected from the group consisting of N, O and S. Any atom of nitrogen and sulfur present in a heterocyclic ring of such a heterocyclic ring structure can be oxidized. Unless otherwise indicated, the terms "heterocyclic ring" or "heterocyclic ring system" includes aromatic rings, as well as non-aromatic rings that may be saturated, partially saturated or completely saturated non-aromatic rings. In addition, unless otherwise indicated the term "heterocyclic ring system" includes ring structures where all rings contain at least one heteroatom as well as structures that have less than all rings in the ring structure containing the less a heteroatom, for example, bicyclic ring structures where a ring is a benzene ring and one of the rings has one or more heteroatoms are included in the term "heterocyclic ring systems" as well as bicyclic ring structures where each one of the two rings have at least one heteroatom. In addition, the ring structures described herein may be attached to one.

or more groups that hang from them through any heteroatom or carbon atom which produces a stable structure. In addition, the term "substituted" means that one or more of the hydrogen atoms in the carbon atom (s) or nitrogen atom (s) of the ring of each of the rings in the ring structures described herein is it may change for one or more of the substituents indicated if such change (s) produced a stable compound. The nitrogen atoms in a ring structure can be quaternized, but such compounds are specifically indicated or are included within the term "a pharmaceutically acceptable salt" for a particular compound. When the total number of O and S atoms in a single heterocyclic ring is greater than 1, it is preferred that such atoms are not adjacent to each other. Preferably, there is no more than 1 O or S atom in the same ring of a given heterocyclic ring structure.

Examples of monocyclic, heterocyclic rings and bicyclic in alphabetical order, are acridinyl, azocinyl, benzimidazolyl, benzofuranyl, benzothiophenyl, benzothiofuranyl, benzoxazolyl, benzthiazolyl, benztriazolyl, benztetrazolyl, benzisoxazolyl, benzisothiazolyl, benzimidazalinilo, carbazolyl, 4aH-carbazolyl, carbolinyl, I cinnolinyl , chromanyl, chromenyl, decahydroquinolinyl, 2H, 6H-1,5,2-dithiazinyl, dihydrofuro [2,3-b] tetrahydrofuran, phenanthridinyl, phenantholinyl, phenazinyl, phenothiazinyl, phenoxyatiinyl, phenoxazinyl, phthalazinurazidyl, furanyl imolinyl, furanyl iminyl imidanyl , imidazolyl, 1H-indazolyl, indolinyl, indolizinyl, indolyl, 3H-indolyl, isobenzofuranyl, isochromanyl, isoindazolyl, isoindolinyl, isoindolyl, isoquinolinyl (benzimidazolyl), isothiazolyl, isoxazolyl, morpholinyl, octaphthiozolyl, octaphthiozolyl-1,2-oxadinozolyl, octaphthiozolyl-1,2-oxadinozyl, octaphthiozolyl, octaphthiozolyl-1,2-naphthyriazole-n-oxadidyloxy-nyl-n-nadyl-n-n-n-n-oxy-n-n-n-o-nyl-n-nazo-n-n-n-n-o-nyl-n-dino-n-n-nazo-n-n-n-n-o-n-o-nin-n-1-O-N-1-O-N-1 oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, oxazolidinyl, or xazolilo, oxazolidinyl, pyrimidinyl, piperazinyl, piperidinyl, pyranyl, pyrazinyl, piroazolidinilo, pyrazolinyl, pyrazolyl, pyridazinyl, pyridooxazole, pyridoimidazole, pyridothiazole, pyridinyl, pyridyl, pyrimidinyl, pyrrolidinyl, pyrrolinyl, 2H-pyrrolyl, pyrrolyl, pteridinyl, purinyl, quinazolinyl, quinolinyl, 4H-quinolizinyl, quinoxalinyl, quinuclidinyl, tetrahydrofuranyl, tetrahydroisoquinolinyl, tetrahydroquinolinyl, 6H-1,2,5-thiadazinyl, 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,2,5-thiadiazolyl, 1,3,4-thiadiazolyl, thiantrenyl, thiazolyl, thienyl, thienothiazolyl, thienoxaxazolyl, thienoimidazolyl, thiophenyl, triazinyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1,2,5-triazolyl, 1, 3,4-triazolyl and xanthenyl. Preferred heterocyclic ring structures include, but are not limited to, pyridinyl, furanyl, thienyl, pyrrolyl, pyrazolyl, pyrrolidinyl, imidazolyl, indolyl, benzimidazolyl, 1H-indazolyl, oxazolinyl or isatinoyl. Also included are fused rings and spiro compounds containing, for example, the heterocyclic structures of previous rings.

As used herein the term "heterocyclic aromatic ring system" has essentially the same definition as for monocyclic and bicyclic ring systems except that at least one ring of the ring system is an aromatic heterocyclic ring or the bicyclic ring has a heterocyclic ring. aromatic or non-aromatic fused to an aromatic carbocyclic ring structure.

The terms "halo" or "halogen" as used herein refer to substituents Cl, Br, F or I. The term "haloalkyl," and the like, refers to an aliphatic carbon radical having at least one hydrogen atom. substituted by an atom of Cl, Br, F or I, including mixtures of different halogen atoms. Trihaloalkyl includes trifluoromethyl and the like as preferred radicals, for example.

The term "methylene" refers to -CH2-.

The term "leaving group" in the definition of L1, L2 and Q includes halogen atoms, substituted or unsubstituted alkoxy groups, substituted or unsubstituted aryloxy groups, substituted or unsubstituted alkylthio groups, substituted or unsubstituted alkylsulfinyl groups, alkylsulfonyl groups substituted or unsubstituted, substituted or unsubstituted alkylsulfonyloxy groups, substituted or unsubstituted arylsulfonyloxy groups, and the like. The halogen atom, alkoxy group, aryloxy group, alkylthio group and alkylsulfinyl group have the same meanings as defined above, respectively, the alkyl part of the alkylsulfonyl group and alkylsulfonyloxy group has the same meaning as the alkyl group defined above, and the aryl part of the arylsulfonyloxy group has the same meaning as the aryl defined above. Examples of the substituent include halogen atoms, alkyl groups, a nitro group, and the like, and the halogen atom has the same meaning as the halogen atom defined above. Other examples include methanesulfonate and p-toluenesulfonate.

The term "pharmaceutically acceptable salts" includes salts of compounds derived from the combination of a compound and an organic or inorganic acid. These compounds are useful both in the form of free base and salt.

In practice, the use of the salt form is equivalent to the use of the base form; Both acid addition and base salts are within the scope of the present invention.

"Pharmaceutically acceptable acid addition salts" refers to salts that maintain biological efficacy and

5 properties of the free bases and which are not biologically or otherwise undesirable, formed with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid and the like, and organic acids such as acetic acid, propionic acid , glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicyclic acid Y

10 similar.

"Pharmaceutically acceptable base addition salts" include those derived from inorganic bases such as sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum and the like salts. Particularly preferred are the ammonium, potassium, sodium, calcium and magnesium salts. Salts derived from pharmaceutically acceptable non-toxic organic bases include salts of primary, secondary and tertiary amines, substituted amines including natural substituted amines, cyclic amines and basic ion exchange resins, such as isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine , 2-diethylaminoethanol, trimetamine, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, hydrabamine, choline, betaine, ethylenediamine, glucosamine, methylglucamine, theobromine, purines, piperizine, piperidine, N-ethylpiperidine, resin resins

20 polyamine and the like. Particularly preferred non-toxic organic bases are isopropylamine, diethylamine, ethanolamine, trimethamine, dicyclohexylamine, choline and caffeine.

The present disclosure also encompasses prodrug derivatives of the compounds contained herein. The term "prodrug" refers to a pharmacologically inactive derivative of a parental drug molecule that requires biotransformation, either spontaneously or enzymatically, in the body to release the active drug. Prodrugs are variations or derivatives of the compounds of this invention that have cleavable groups under metabolic conditions. Prodrugs become the compounds of the invention that are pharmaceutically active in vivo, when they undergo sololysis under physiological conditions or undergo enzymatic degradation. The prodrug compounds of this invention can be called single, double, triple, etc., depending on the number of biotransformation steps required to release the active drug in the organism and indicate the number of functionalities present in the precursor type form. Prodrug forms often offer advantages of solubility, tissue compatibility or delayed release in mammalian organisms (see, Bundgard, Design of Prodrugs, pp. 7-9, 21-24, Elsevier, Amsterdam 1985 and Silverman, The Organic Chemistry of Drug Design and Drug Action, pp. 352-401, Academic Press, San Diego, Calif., 1992). The

Prodrugs commonly known in the art include acid derivatives that are well known to those skilled in the art, such as, for example, esters prepared by reacting parental acids with a suitable alcohol, or amides prepared by reacting the parent acid compound with an amine. , or basic groups that have been reacted to form an acyl base derivative. In addition, the prodrug derivatives of this invention can be combined with other features taught herein to increase bioavailability.

"Biological property" for the purposes of this document means an in vivo effect or antigenic function or activity that is performed directly or indirectly by a compound of this invention that are often shown by in vitro assays. Effector functions include receptor or ligand binding, any enzymatic activity or enzyme modulating activity, any transporter binding activity, any activity

Hormonal, any activity in promoting or inhibiting the adhesion of cells to an extracellular matrix or cell surface molecules, or any structural role. Antigenic functions include possession of an epitope or antigenic site that is capable of reacting with antibodies produced against it.

The present disclosure relates to a process for making nitrogen-containing heterocyclic compounds.

image7

where

R1 is a member selected from the group consisting of: -CN, -O-C1-8 alkyl which is straight or branched chain; -O-phenyl, -O-naphthyl, -O-indolyl and -O-isoquinolyl; R2 and R4 are each independently a member selected from the group consisting of: hydrogen, -O-CH3, -O (-CH2) -CH3, -O (-CH2) 2-CH3, -O-CH2-CH = CH2, -O-CH2-C≡CH and -O (-CH2) n-R3; where one of the R2 and R4 groups is -O (-CH2) n-R3 and the remaining R2 or R4 group is different from -O (-CH2) n-R3; n is from 2 to 5; R3 is a member selected from the group consisting of:

image8

10 and a saturated 4- or 10-membered mono or bicyclic heterocyclic ring system, partially unsaturated or completely unsaturated having at least one nitrogen atom and 0 to 3 additional heteroatoms selected from the group consisting of O, N and S, in where the ring system may be unsubstituted or may be substituted by 1 to 4 members selected from the group consisting of H, halo, halo-lower alkyl, lower alkyl, lower alkynyl, lower acyl, lower alkoxy, hydroxy, nitro, amino and the like, where the ring system

15 may be directly linked to the adjacent methylene group or may be linked through an ether bond, and all isomers, salts, hydrates, solvates and pharmaceutically acceptable prodrug derivatives thereof.

A preferred process of the present disclosure is a process for the production of such compounds wherein R3 is a member selected from the group consisting of:

image9

A particularly preferred process of the present disclosure is a process for making compounds according to formula A above wherein R1 is a member selected from the group consisting of CN, -O-methyl, -O-ethyl, -Opropyl, -O- isopropyl, -O-butyl, -O-butyl, -O-isoamyl, 1-naphthyloxy, 2-naphthyloxy, 4-indolyloxy, 5-indolyloxy, 5-isoquinolyloxy, and positional isomers and homologs thereof, and all Isomers, salts, hydrates, solvates and pharmaceutically acceptable prodrug derivatives of such compounds. More preferred is a process disclosed to do

Compounds wherein n is 2 or 3 and R1 is -O-isopropyl or CN and R3 is a cyclic amine.

The disclosed process also provides pharmaceutically acceptable salts of the compounds according to the formula.

(A) which include acid addition salts, metal salts, ammonium salts, organic amine addition salts, pharmaceutically acceptable amino acid addition salts etc. Examples of pharmaceutically acceptable acid addition salts of the compounds of formula (A) are inorganic acid addition salts such as hydrochloride, sulfate and phosphate, and organic acid addition salts such as acetate, maleate, fumarate, tartrate, citrate. and methanesulfonate. Examples of pharmaceutically acceptable metal salts are alkali metal salts such as sodium salt and potassium salt, alkaline earth metal salts such as magnesium salt and calcium salt, aluminum salt and zinc salt. Examples of pharmaceutically acceptable ammonium salts are salt of

40 ammonium and tetramethylammonium salt. Examples of pharmaceutically acceptable organic amine addition salts include salts of heterocyclic amines such as morpholine and piperidine salts. Examples of pharmaceutically acceptable amino acid addition salts are salts with lysine, glycine and phenylalanine. The process also envisages making pharmaceutically acceptable isomers, hydrates, solvates and prodrug derivatives of the compounds according to formula (A) and would be apparent to the person skilled in the art.

The process now disclosed can be easily adapted to make other compounds in the art, such as the nitrogen heterocyclic compounds described in WO 98/14431 published April 9, 1998. Accordingly, the present disclosure also provides for making such compounds using the present process or easily apparent variations thereof.

In a preferred embodiment the disclosure provides a process for making compounds according to the

image10

Formula A (1) Formula A (2)

Wherein R1 is a member selected from the group consisting of: -CN, -O-C1-8 alkyl which is straight or branched chain; -O-phenyl, -O-naphthyl, -O-indolyl and -O-isoquinolyl; and all isomers, salts, hydrates, solvates and pharmaceutically acceptable prodrug derivatives thereof.

A particularly preferred process of the present disclosure is a process for making compounds according to formula A (1) or A (2) above wherein R1 is -O-isopropyl or CN, n is 2 or 3 and R3 is a cyclic amine , as well as positional isomers and homologs thereof, and all isomers, salts, hydrates, solvates and pharmaceutically acceptable prodrug derivatives of such compounds.

A more preferred process of the disclosure is a process for making compounds according to formula A (1) or A (2) above wherein n is 3, R1 is -O-isopropyl or CN and R3 is a member selected from the group that consists in:

30 and all isomers, salts, hydrates, solvates and pharmaceutically acceptable prodrug derivatives thereof.

Additionally preferred is such a disclosure process where n is 3, R1 is -O-isopropyl or CN and R3 is a

image11

image12

image13

same.

Most preferred is a process of disclosure to make compounds of formula A (1) or A (2) wherein n is 3, R1 is -O-isopropyl or CN and R3 is N-piperidine or N-pyrrolidine.

Pharmaceutically acceptable salts of the compounds according to formula (A) include acid addition salts, metal salts, ammonium salts, organic amine addition salts, pharmaceutically acceptable amino acid addition salts etc., 10.

The process now disclosed is not limited by the compounds listed above, but includes intermediates to make such compounds or other related compounds. Analogs of bicyclic compounds are contemplated and would be apparent to the person skilled in the art.

The compounds can be prepared using methods and procedures in general as described below, however other leaving groups can be used.

The present disclosure is directed to a process for preparing a nitrogen-containing heterocyclic compound of

image14

which includes the steps of:

(a) etherifying the hydroxy group of a compound of formula I or its position isomer with a compound of formula II

Wherein L1 is a leaving group such as Cl and the like, which is less reactive than an leaving group forming ether L2 such as Br and the like, under basic etherification conditions, preferably where the base is preferably potassium carbonate, carbonate of sodium, sodium hydroxide and the like, in the presence of an appropriate solvent such as toluene, methanol, ethanol, ether, THF and the like, preferably ethanol or toluene, at reflux temperature of the solvent about 2-6 hours, preferably about 3 to 4 hours to produce

image15

(b) nitrate a compound according to formula III, or its position isomer, to give a compound according to formula IV, or its position isomer, preferably at a temperature from about 0 ° C to 80 ° C, preferably

About 0 to 20 ° C in nitric acid and an appropriate solvent such as a mixture of acetic acid and dichloromethane, as follows:

image 1

(c) reacting the compound of formula IV, or its position isomer, with an amine-containing compound for the appropriate R3 group, such as a piperidine, pyrrolidine, morpholine, piperazine, 4-methylpiperidine or 2-methylpiperidine, in the presence of a basic catalyst such as potassium carbonate, sodium carbonate, hydroxide of

5 sodium, and the like, preferably potassium carbonate and sodium iodide and a solvent such as toluene, ethanol, THF, ether, glyme (1,2-dimethoxyethane), diglyme, MTBE, or the like, to replace the L1 group with a R3 group, and provide a compound of formula V, or its position isomer as follows: image 1

(d) reduce the nitro group in the compound of formula V, or in its position isomer, to an amino group and thereby produce a compound of formula VI, or its position isomer, as follows: image 1

(and)

reacting the compound of formula V, or its position isomer, in the form of ammonium and formamide at about 120 ° C to 140 ° C, preferably, at about 130 ° C to produce a cyclic derivative of quinazoline of formula VII, or its position isomer, as follows:

(F)

 replacing the hydroxy group of compound VII, or its position isomer, with a leaving group Q, preferably Q is the leaving group bromine, chlorine, p-toluenesulfonate, methylsulfonate and the like, preferably chlorine derived from a chlorinating agent such as chloride thionyl, to provide a compound of formula VIII, or its position isomer, as follows:

(g)

reacting the compound of formula VIII, or its position isomer, with a compound containing an amino group of formula IX to replace leaving group Q and providing a compound of formula X, or its position isomer, as follows:

(h)

and optionally, producing a salt, such as the halohydrate salt, of the compound of formula X, or its position isomer, as follows:

image16

image17

image 1

image 1

and all isomers, salts, hydrates, solvates and pharmaceutically acceptable prodrug derivatives thereof. The disclosure also provides a process for preparing an intermediate compound having the formula VIII

image18

Wherein n, R3 and R4 are defined as above, and Q is a leaving group different from a hydroxyl group, which can be substituted by an amino group or other intermediate group that is subsequently replaced by an amino group, or a salt of the same.

In the above process, leaving groups such as halogen, lower alkoxy, lower alkylthio, lower alkylsulfonyloxy, arylsulfonyloxy, etc., may be used, when necessary except for the reaction point, followed by deprotection. The amino protecting groups are, for example those described in TW Greene, Protective Groups in Organic Synthesis, John Wiley & Sons Inc. (1981), etc., such as ethoxycarbonyl, t-butoxycarbonyl, acetyl, benzyl and the like that would be apparent for The subject matter expert. Protective groups can be entered and

20 remove according to conventional methods used in organic synthetic chemistry, for example, T. W. Greene, Protective Groups in Organic Synthesis, John Wiley & Sons Inc. (1981).

Appropriate solvents include a lower alcohol, such as methanol, ethanol, isopropanol, etc., a halogenated hydrocarbon, such as chloroform, dichloromethane, etc., an aromatic hydrocarbon, such as benzene, toluene, etc., a

Ether solvent, such as diethyl ether, THF, 1,4-dioxane, etc., a polar aprotic solvent, such as dimethylformamide, N-methylpyrrolidone, dimethylsulfoxide, pyridine, etc., or a solvent thereof, optionally in presence of a base. Examples of bases include organic bases, such as triethylamine, pyridine, etc., inorganic bases, such as potassium carbonate, sodium carbonate, sodium hydroxide, sodium hydride, etc., metal alkoxides, such as sodium methoxide. , potassium tert-butoxide, etc., and the like.

In such processes, if the defined groups change under the conditions of the working method or are not suitable for carrying out the method, the desired compound can be obtained using the methods for introducing and removing protective groups that are conventionally used in chemistry. organic synthetic. See, for example, T. W. Greene, Protective Groups in Organic Synthesis, John Wiley & Sons Inc. (1981), etc. The conversion of functional groups contained in the substituents can be carried out by known methods. See, for example, R.

C. Larock, Comprehensive Organic Transformations (1989), in addition to the processes described above, and some of the active compounds of formula I can be used as intermediates for further synthesis of new derivatives according to formula A.

The intermediates and compounds desired in the processes described above can be isolated and purified by purification methods conventionally used in organic synthetic chemistry, for example, neutralization, filtration, extraction, washing, drying, concentration, recrystallization, and various types of chromatography. The intermediates can be subjected to the subsequent reaction without purification.

There may be tautomers for some of formula A, and the present invention covers all possible isomers including tautomers and mixtures thereof, the process of making them would be apparent to the person skilled in the art. When chiral carbons lend themselves to two different enantiomers, both enantiomers are contemplated as well as the procedures for separating the two enantiomers. In the compounds of this invention, carbon atoms attached to four non-identical substituents are asymmetric. According to this, the compounds may also exist as diastereomers, enantiomers or mixtures thereof. The syntheses described herein may employ racemates, enantiomers or diastereomers as starting materials or intermediates. The diastereomeric products resulting from such synthesis can be separated by chromatographic or crystallization methods, or by other methods known in the art. Also, mixtures of enantiomeric products can be separated using the same techniques or by other methods known in the art. Each of the asymmetric carbon atoms, when present in the compounds of this invention, can be in one of two configurations (R or S) and both are within the scope of the present invention. In the processes described above, the final products may, in some cases, contain a small amount of diastereomeric or enantiomeric products, however, these products do not affect their therapeutic or diagnostic application.

In the case where a salt of a compound of formula A is desired and the compound is produced in the form of the desired salt, it can be subjected to purification as such. In the case where a compound of formula A is produced in the free state and its salt is desired, the compound of formula A is dissolved or suspended in a suitable organic solvent, followed by the addition of an acid or a base to form a salt. Preferably the solvent for recrystallization and salt formation is a lower alcohol, preferably methanol or ethanol.

The following examples are provided to illustrate the invention. The examples are not intended to limit the scope of the present invention and should not be so interpreted. The amounts are given in parts by weight or percentages by weight unless otherwise indicated. All patents and publications cited are incorporated herein by reference. The following specific examples are provided to better assist the reader in various aspects of the practice of the present invention. Since these specific examples are merely illustrative, nothing in the following description should be construed as limiting the invention in any way.

Scheme 1

image 1

5 Such examples of the process according to the invention are only an illustration of a preferred aspect of the invention. Other procedures and adaptations will be apparent to the person skilled in the art after examining these reaction schemes and the structures of the compounds according to the invention. Such procedures are considered to be within the scope of the present invention.

In addition, the compounds of formula A and pharmaceutically acceptable salts thereof may exist in the form of adducts with water (hydrates) or various solvents, which are also within the scope of the present invention.

The following non-limiting examples are provided to better illustrate the present invention.

Example 1: Preparation of 4- [6-Methoxy-7- (3-piperidin-1-yl-propoxy) quinazolin-4-yl] -piperazine-1-carboxylic acid 4- (isopropoxy-phenyl) -amide

Step 1

In a round bottom flask, 1-chloro-3-bromopropane (1.28 mol) was charged followed by a solution of aqueous potassium carbonate, ethyl vanillate (0.51 mol) and N-butylammonium bromide (0.0255 moles) and the resulting reaction mixture was heated at about 70 to about 100 ° C for 0.5-4 hours until termination of the reaction to compound III was confirmed by HPLC / CCF analysis. The reaction mixture was cooled to about 20-25 ° C and dichloromethane was added. The resulting biphasic mixture was separated. The organic phase was washed with water after brine solution and the solvent was removed in vacuo to about 1/5 of its original volume. This solution of compound III in dichloromethane was taken to step # 2.

Step 2

In a round bottom flask equipped with a condenser, thermometer and mechanical stirrer, the solution of III in dichloromethane was charged followed by acetic acid (0.5 L) and the resulting light brown solution was cooled to about 0-5 ° C. The 70% nitric acid (1.53 mol) was charged dropwise to the rapid stirring solution for about 40-60 minutes. The resulting light brown solution was slowly heated to about 50-70 ° C and allowed to stir at this temperature for about 2-10 hours until the completion of the reaction was confirmed by HPLC / CCF analysis. The orange solution was poured into ice / water (1.0 L) and dichloromethane (0.5 L). The solution was allowed to warm to about 20 ° C, the phases were separated and the organic phase was washed several times with deionized water followed by brine. The solvent was removed under reduced pressure to approximately 1/5 of the original volume at which time ethanol was introduced. The ethanolic solution was allowed to cool to about 20 ° C for 10-16 hours, then cooled further to about 0-10 ° C for about 1-3 hours. The off-white solid was collected by vacuum filtration to give about 82% (based on the starting weight of I) of IV. Product identity was confirmed by proton NMR, carbon-13 and mass spectrometry analysis.

Step 3

IV (0.31 mol) was charged to a round bottom flask followed by toluene (500 ml), an aqueous solution of potassium carbonate, N-butylammonium bromide (0.0155 mol), sodium iodide (0.62 moles) and piperidine (0.93 moles) and the resulting reaction mixture was heated at about 60-100 ° C for about 1-10 hours until the completion of the reaction was confirmed by HPLC / CCF analysis. The reaction mixture was cooled to about 20-25 ° C and the phases separated. The aqueous phase was extracted once with toluene. The combined organic phases were washed with water, 3% thiosulfate solution and then brine. The toluene was removed under reduced pressure to approximately 1/5 of its original volume at which time ethanol (500 ml) and water (200 ml) were added and V was taken in solution to step # 4.

Step 4

In a round bottom flask containing the solution of V in ethanol / water / toluene, palladium on carbon catalyst (50% wet) was charged and the reaction mixture was heated to about 40-50 ° C. To this heated reaction mixture, a solution made in advance of potassium format (0.62 mol) and formic acid (0.93 mol) was charged in deionized water for about 0.5-3 hours maintaining the temperature of the solution between 40-55 ° C and the pH of the solution between 3-6. The reaction mixture was allowed to stir for about 0.5-4 hours at about 40-50 ° C at which time the HPLC / CCF analysis confirmed the completion of the reaction. The reaction mixture was filtered through celite. The filtrate was evaporated at atmospheric pressure until a vessel temperature of about 80-90 ° C was reached. The remaining aqueous solution was cooled to about 20-25 ° C at which time ethyl acetate (600 ml) was charged followed by an aqueous solution of potassium carbonate sufficient to adjust the pH to> 10. After vigorous stirring the phases were separated, the basic aqueous phase was extracted with ethyl acetate and the combined organic phases were washed with water followed by brine solution. This ethyl acetate solution containing VI was taken directly to step # 5.

Step 5

A round bottom flask containing VI (0.31 mol) in ethyl acetate and one equipped with a mechanical stirrer, thermometer and distillation apparatus were charged with formamide (210 ml) and ethyl acetate distillation was started until it was It reached a vessel temperature of around 120-130 ° C. The reaction mixture was cooled to about 60-80 ° C at which time ammonium format (0.37 mol) was charged. The reaction mixture was further heated to about 110-150 ° C for about 2-12 hours, preferably at about 130 ° C for about six hours, until the completion of the reaction was confirmed by HPLC / CCF analysis. The reaction mixture was cooled to about 20-25 ° C at which time they were charged in digitimate succession (800 ml) followed by methyl-t-butyl ether (260 ml). The suspension was allowed to stir at about 2025 ° C for about 1-10 hours, preferably 3 hours, and then cooled to about 0-10 ° C for about 1-3 hours. The product was filtered, the cake was washed with MTBE. The wet cake was transferred to another round bottom flask and MTBE (800 ml) was charged to the white product and the suspension was vigorously stirred for about 1-4 hours at about 20-25 ° C. The product was filtered and washed with MTBE and then dried at a constant weight at about 30-50 ° C under vacuum to yield 70% (based on starting weight of IV) of VII. Product identity was confirmed by proton NMR, C-13 and mass spectrometry analysis.

Step 6 In a round bottom flask equipped with a mechanical stirrer, thermometer, condenser and nitrogen purge, toluene (500 ml) was loaded followed by VII (0.28 mol) and the resulting suspension was cooled to about 0-10 ° C at which time which was charged with thionyl chloride (500 ml) in portions for several hours while maintaining the temperature of the solution of at least <25 ° C. The reaction mixture was cooled again to about 1015 ° C at which time dimethylformamide (100 ml) was charged portionwise for several hours while maintaining a dissolution temperature of at least <35 ° C. The reaction mixture was heated to about 80-85 ° C where it was maintained for about 1-5 hours until the completion of the reaction was confirmed by HPLC / CCF analysis. Toluene (500 ml) was charged to the reaction mixture while maintaining the temperature during the addition to at least> 50 ° C. The reaction mixture was allowed to cool to about 20-25 ° C and was maintained at this temperature for about 8-16 hours. The resulting precipitated solid was filtered and the cake was washed with toluene. The cake still wet with toluene was added portionwise to a solution prepared in advance of 20% potassium bicarbonate and dichloromethane which was cooled to about 0-5 ° C. After ensuring that a solution pH of at least> 10 was obtained, the phases were separated and the aqueous one was extracted once more with dichloromethane. The combined organic phases were washed with water and then brine. The dichloromethane solution was evaporated under reduced pressure at a temperature of at least <40 ° C to about 1/5 of its original volume at which time acetonitrile (1800 ml) was charged. The resulting precipitated white solid was cooled to about 20-25 ° C and allowed to stir at this temperature for about 4 hours. The product was collected by filtration and washed with acetonitrile to give after drying at a constant weight under vacuum at about 35-50 ° C, a yield of 72% of at least> 95% of VIII. The identity of this product was confirmed by proton, C-13 and mass spectrometry analysis.

Step 7

A round bottom flask equipped with a mechanical stirrer, thermometer, reflux condenser and nitrogen purge was charged VIII (0.298 moles), dimethylformamide (800 ml), potassium carbonate (0.746 moles) and IX (0.300 moles) and the mixture The reaction was stirred at about 20-50 ° C for about 2-24 hours, preferably about 6 hours at about 40 ° C, until the completion of the reaction was confirmed by HPLC / CCF analysis. The reaction mixture was cooled to about 20-25 ° C at which time it was charged to a solution of deionized water and dichloromethane. The phases were separated and the aqueous phase was extracted with dichloromethane. The organic phases were combined and washed with 20% sodium chloride solution, 20% ammonium chloride solution followed by brine. Dichloromethane (DCM) was removed to approximately 1/5 of its original volume by evaporation under reduced pressure at a temperature of at least <40 ° C. Ethanol (1000 ml) was charged to this DCM solution and the solution was heated to about 40-50 ° C at which time coal was added. The reaction mixture was stirred at about 40-50 ° C for about 0.5-1.0 hours and then filtered on celite to remove carbon. The filtrate was reheated to about 40 ° C at which time a solution prepared in advance of sulfuric acid in ethanol was added in portions until a solution pH of about 2-4 was reached. The resulting white suspension was cooled to about 20-25 ° C and stirred at this temperature for about 2-8 hours. It was further cooled to about 0-10 ° C where it was stirred for about 1-3 hours. The product was collected by filtration and the cake was washed with ethanol of about 0-10 ° C. The material was dried under vacuum at about 40-50 ° C until a constant weight was obtained to give a yield of 80% X with at least> 90% purity by HPLC analysis.

Step 8

In a round bottom flask equipped with a mechanical stirrer, condenser, thermometer and argon purge, crude X (0.167 mol) was charged followed by ethanol (600 ml) and deionized water (200 ml). The reaction mixture was heated to about 55-60 ° C at which time the total solution was reached. The solution was cooled to about 20-25 ° C for at least 1-4 hours, preferably about 3 hours. The reaction mixture was further cooled to about 0-5 ° C where it was maintained for about 1-3 hours. The product was collected by filtration, the cake was washed with ethanol of about 0-5 ° C and then dried under vacuum at about 35-55 ° C, preferably around 45 ° C, until XI was obtained with an LDD <1%. Compound XI was isolated in 83% yield and determined to be at least> 99% pure by HPLC analysis without impurities of at least> 0.5%. The identity of the compound was confirmed by comparison to a previously synthesized and fully characterized analytical reference standard.

Example 2: Preparation of 4- [6-methoxy-7- (3-morpholin-4-yl-propoxy) quinazolin-4-yl] -piperazine-1-carboxylic acid (4-isopropoxy-phenyl) -amide

Step 1

In a round bottom flask, 1-chloro-3-bromopropane (1.53 mol) was charged followed by a solution of aqueous potassium carbonate, ethyl vanillate (0.51 mol) and N-butylammonium bromide (0.0255 mol ) and the resulting reaction mixture was heated at about 70-100 ° C for about 0.5-4 hours until the termination of the reaction of the title compound was confirmed by HPLC / CCF analysis. The reaction mixture was cooled to about 20-25 ° C and dichloromethane (500 ml) was added and the resulting biphasic mixture was separated. The organic phase was washed with water after brine solution and the solvent was removed in vacuo to about 1/5 of its original volume. This solution of compound III in dichloromethane was taken to step # 2.

Step 2

In a round bottom flask equipped with a condenser, thermometer and mechanical stirrer, the solution of III (0.51 mol) in dichloromethane was charged followed by acetic acid (500 ml) and the resulting light brown solution was cooled to about 0-5 ° C . The rapid stirring solution was charged dropwise 70% nitric acid (1.53 mol) for about 40-60 minutes. The resulting light brown solution was slowly heated to about 50-70 ° C and allowed to stir at this temperature for about 2-10 hours until the completion of the reaction was confirmed by HPLC / CCF analysis. The red solution was poured into ice / water (1000 ml) and dichloromethane (500 ml). The solution was allowed to warm to about 20 ° C, the phases were separated and the organic phase was washed several times with deionized water followed by brine. The solvent was removed under reduced pressure to approximately 1/5 of the original volume at which time ethanol was introduced. The solution was allowed to cool to about 20 ° C for about 10-16 hours, then cooled further to about 0-10 ° C for about 1-3 hours. The off-white solid was collected by vacuum filtration to give 82% (based on the starting weight of I) of IV. Product identity was confirmed by proton NMR, carbon13 and mass spectrometry analysis.

Step 3

IV (0.315 mol) was charged into a round bottom flask followed by toluene (500 ml), an aqueous solution of potassium carbonate, N-butylammonium bromide (0.0158 mol), sodium iodide (0.48 mol) and morpholine (0.945 mol) and the resulting reaction mixture was heated at about 60-100 ° C for about 1-10 hours until the termination of the reaction was confirmed by HPLC / CCF analysis. The reaction mixture was cooled to about 20-25 ° C and the phases separated. The aqueous phase was extracted once with toluene. The combined organic phases were washed with water, 3% thiosulfate solution and then brine. The toluene was removed under reduced pressure to approximately 1/5 of its original volume at which time ethanol (500 ml) and water (200 ml) were added and V was taken in solution to step # 4.

Step 4

In a round bottom flask containing the solution of V in ethanol / water / toluene, palladium on carbon catalyst (50% wet) was charged and the reaction mixture was heated to about 40-50 ° C. To this heated reaction mixture was loaded a solution made in advance of potassium format (0.63 mol) and formic acid (0.945 mol) in deionized water for about 0.5-3 hours keeping the temperature of the solution between 40 -55 ° C and the pH of the solution between at least 3-6. The reaction mixture was allowed to stir for about 0.5-4 hours at about 40-50 ° C at which time the HPLC / CCF analysis confirmed the completion of the reaction. The reaction mixture was filtered through celite. The filtrate was evaporated at atmospheric pressure until a vessel temperature of about 80-90 ° C was reached. The remaining aqueous solution was cooled to about 20-25 ° C at which time ethyl acetate (500 ml) was charged followed by an aqueous solution of potassium carbonate sufficient to adjust the pH to at least> 10. After vigorous stirring the phases were separated, the basic aqueous phase was extracted with ethyl acetate and the combined organic phases were washed with water followed by brine solution. This ethyl acetate solution containing VI was taken directly to step # 5.

Step 5

A round bottom flask containing VI (0.315 mol) in ethyl acetate and one equipped with a mechanical stirrer, thermometer and distillation apparatus were charged with formamide (210 ml) and ethyl acetate distillation was started until a vessel temperature of about 120-130 ° C. The reaction mixture was cooled to about 60-80 ° C at which time ammonium format (0.388 mol) was charged. The reaction mixture was further heated to about 110-150 ° C for about 2-12 hours, preferably at about 130 ° C for about six hours, until the completion of the reaction was confirmed by HPLC / CCF analysis. The reaction mixture was cooled to about 20-25 ° C at which time they were charged in digitimate succession (800 ml) followed by methyl-t-butyl ether (225 ml). The suspension was allowed to stir at about 2025 ° C for about 1-10 hours, preferably 3 hours, and then cooled to about 0-10 ° C for about 1-3 hours. The product was filtered, the cake was washed with MTBE. The wet cake was transferred to another round bottom flask and MTBE (800 ml) was charged to the white product and the suspension was vigorously stirred for about 1-4 hours at about 20-25 ° C. The product was filtered and washed with MTBE and then dried at a constant weight at about 30-50 ° C under vacuum to give a yield of at least 72% (based on starting weight of IV) of VII. Product identity was confirmed by proton NMR, C-13 and mass spectrometry analysis.

Step 6 In a round bottom flask equipped with a mechanical stirrer, thermometer, condenser and nitrogen purge, toluene (500 ml) was loaded followed by VII (0.278 moles) and the resulting suspension was cooled to about 0-10 ° C at which time thionyl chloride (500 ml) was charged in portions for several hours while maintaining the solution temperature of at least <25 ° C. The reaction mixture was cooled again to about 1015 ° C at which time dimethylformamide (100 ml) was charged portionwise for several hours while maintaining a dissolution temperature of at least <35 ° C. The reaction mixture was heated to about 80-85 ° C where it was maintained for about 1-5 hours until the completion of the reaction was confirmed by HPLC / CCF analysis. Toluene (500 ml) was charged to the reaction mixture while maintaining the temperature during the addition to at least> 50 ° C. The reaction mixture was allowed to cool to about 20-25 ° C and was maintained at this temperature for about 8-16 hours. The resulting precipitated solid was filtered and the cake was washed with toluene. The cake still wet with toluene was added portionwise to a solution prepared in advance of 20% potassium bicarbonate and dichloromethane which was cooled to about 0-5 ° C. After ensuring that a solution pH of at least> 10 was obtained, the phases were separated and the aqueous one was extracted once more with dichloromethane. The combined organic phases were washed with water and then brine. The dichloromethane solution was evaporated under reduced pressure at a temperature of at least <40 ° C to about 1/5 of its original volume at which time acetonitrile (1400 ml) was charged. The resulting precipitated white solid was cooled to about 20-25 ° C and allowed to stir at this temperature for about 4 hours. The product was collected by filtration and washed with acetonitrile to give after drying at a constant vacuum weight at about 35-50 ° C, a yield of about 75% of at least> 95% VIII. The identity of this product was confirmed by proton, C-13 and mass spectrometry analysis.

Step 7

A round bottom flask equipped with a mechanical stirrer, thermometer, reflux condenser and nitrogen purge was charged VIII (0.298 moles), dimethylformamide (800 ml), potassium carbonate (0.745 moles) and IX (0.300 moles) and the mixture The reaction was stirred at about 20-50 ° C for about 2-24 hours, preferably about 6 hours at about 40 ° C, until the completion of the reaction was confirmed by HPLC / CCF analysis. The reaction mixture was cooled to about 20-25 ° C at which time it was charged to a solution of deionized water and dichloromethane. The phases were separated and the aqueous phase was extracted with dichloromethane. The organic phases were combined and washed with 20% sodium chloride solution, 20% ammonium chloride solution followed by brine. Dichloromethane was removed to approximately 1/5 of its original volume by evaporation under reduced pressure at a temperature of at least <40 ° C. Ethanol (800 ml) was charged to this DCM solution and the solution was heated to about 40-50 ° C at which time coal was added. The reaction mixture was stirred at about 40-50 ° C for about 0.5-1.0 hours and then filtered on celite to remove carbon. The filtrate was reheated to about 40 ° C at which time a solution prepared in advance of sulfuric acid in ethanol was added in portions until a solution pH of at least about 2-3 was reached. The resulting white suspension was cooled to about 20-25 ° C and stirred at this temperature for about 2-8 hours. It was further cooled to about 0-10 ° C where it was stirred for about 1-3 hours. The product was collected by filtration and the cake was washed with ethanol of about 0-10 ° C. The material was dried under vacuum at about 40-50 ° C until a constant weight was obtained to give a yield of 83% X with at least> 90% purity by HPLC analysis.

Step 8

In a round bottom flask equipped with a mechanical stirrer, condenser, thermometer and argon purge, crude X (0.166 mol) was charged followed by methanol (600 ml) and deionized water (40 ml). The reaction mixture was heated to about 50-55 ° C at which time the total solution was reached. The solution was cooled to about 20-25 ° C for about 1-4 hours, preferably about 3 hours. The reaction mixture was further cooled to about 0-5 ° C where it was maintained for about 1-3 hours. The product was collected by filtration and the cake was washed with methanol of about 0-5 ° C and then dried under vacuum at about 35-55 ° C, preferably around 45 ° C, until XI was obtained with an LDD <1%. Compound XI was isolated in a yield of about 75% and determined to be at least> 99% pure by HPLC analysis without impurities of at least> 0.5%. The identity of the compound was confirmed by comparison to a previously synthesized and fully characterized analytical reference standard.

It is preferred to use the route of administration that is most effective for treatment. For example, administration is done orally or non-orally by intrarectal, intraoral, subcutaneous, intramuscular or intravenous administration.

Examples of the forms for administration are capsules, tablets, granules, powders, syrups, emulsions, suppositories and injections.

Liquid compositions such as emulsions and syrups that are suitable for oral administration can be prepared using water, sugars such as sucrose, sorbitol and fructose, glycols such as polyethylene glycol and propylene glycol, oils such as sesame oil, olive oil and olive oil. soy, preservatives such as benzoates, flavorings such as strawberry and mint flavor, etc.

Capsules, tablets, powders and granules can be prepared using excipients such as lactose, glucose, sucrose and mannitol, disintegrating agents such as starch and sodium alginate, lubricants such as magnesium stearate and talc, binders such as polyvinyl alcohol, hydroxypropylcellulose and gelatin. , surfactants such as fatty acid esters, plasticizers such as glycerin, etc.

Compositions for topical application are prepared by dissolving or suspending an active ingredient in one or more types of solvents such as mineral oil, petrolatum and polyhydric alcohol, or other bases used for topical drugs.

Compositions for intestinal administration are prepared using usual supports such as cocoa fat, hydrogenated fat and carboxylic acid of hydrogenated fat, and are supplied as suppositories.

Compositions suitable for non-oral administration preferably comprise a sterile aqueous preparation containing an active ingredient that is isotonic with the blood of the recipient. For example, injections are prepared using a support comprising a saline solution, a glucose solution, or a mixture of a saline solution and a glucose solution. Compositions for non-oral administration may also be formulated to contain one or more types of additives selected from glycols, oils, flavorings, preservatives (including antioxidants), excipients, disintegrating agents, lubricants, binders, surfactants and plasticizers that are used for Preparation of compositions for oral administration.

The effective dose and administration schedule for each of the compounds of formula (A) or a pharmaceutically acceptable salt thereof will vary depending on the route of administration, the age and body weight of the patient, and the type and degree of the disease to be treated. However, it is generally appropriate to administer a compound of formula (A) or a pharmaceutically acceptable salt thereof in a dose of 0.011000 mg / adult / day, preferably 5-500 mg / adult / day, in one or more parts

All the compounds of the present invention produced by the process according to the invention can be immediately applied to the treatment of mammalian kinase dependent diseases such as kinase inhibitors, specifically those related to tyrosine kinases. Specifically, compounds having an IC50 in the range from about 10 nM to about 10 µM are preferred. Even more preferred are compounds that have an IC50 in the range from about 10 nM to about 1 µM. Most preferred with compounds having an IC50 value that is less than about 1 µM. Specific compounds of the present invention having an activity to specifically inhibit one of three types of protein kinases (for example, phosphorylating tyrosine kinases, phosphorylating tyrosine and threonine kinases and phosphorylating threonine kinases) can be selected. Tyrosine kinase dependent diseases include hyperproliferative malfunction that is caused or maintained by abnormal tyrosine kinase activity.

Examples include psoriasis, pulmonary fibrosis, glomerulonephritis, cancers, atherosclerosis and antiangiogenesis (for example, tumor proliferation or diabetic retinopathy). Although the relationships of other classes of kinases with specific diseases are not well known, a selective compound that inhibits tyrosine kinase is considered to have a useful therapeutic effect. In addition, it is understood that other classes of kinases have their own useful therapeutic effects. Quercetin, genistein and staurosporine, which are inhibitors of tyrosine kinases inhibit many other types of protein kinases in addition to tyrosine kinase, and have a strong cytotoxicity as a result of the lack of specificity for them. Accordingly, tyrosine kinase inhibitors (or other kinase inhibitors) that have a tendency to induce undesirable side effects due to lack of selectivity can be identified using a usual test to measure cytotoxicity.

The present invention provides a process for making nitrogen-containing heterocyclic compounds and pharmaceutically acceptable salts thereof that inhibit phosphorylation of the PDGF receptor to prevent abnormal cell growth and are therefore useful for the prevention or treatment of proliferative cell diseases. such as arteriosclerosis, vascular resorption, cancer and glomerulosclerosis. Other variations in the processes and compounds according to the invention will be apparent when considering the preferred embodiments of the present invention and are contemplated herein as being within the scope of the present invention.

Compositions or formulations of the compounds of the invention are prepared for storage or administration by mixing the compound having the desired degree of purity with physiologically acceptable carriers, excipients, stabilizers, etc. and can be provided in sustained release or dependent release formulations. weather. Acceptable carriers or diluents for therapeutic use are well known in the pharmaceutical field, and are described, for example, in Remington’s Pharmaceutical Sciences, Mack Publishing Co.,

(A.R. Gennaro edit. 1985). Such materials are not toxic to the receptor at the doses and concentrations employed, and include buffers such as phosphate, citrate, acetate and other salts of organic acids, antioxidants such as ascorbic acid, low molecular weight peptides (less than about ten residues ) such as polyarginine, proteins, such as serum albumin, gelatin or immunoglobulins, hydrophilic polymers such as polyvinylpyrrolidone, amino acids, such as glycine, glutamic acid, aspartic acid or arginine, monosaccharides, disaccharides and other carbohydrates including cellulose or its derivatives, glucose , mannose or dextrins, chelating agents such as EDTA, polyols such as mannitol or sorbitol, counterions such as sodium and / or non-ionic surfactants such as TWEEN®, PLURONICS® or polyethylene glycol.

The term "effective amount" is an amount necessary to administer the compound according to the present invention to provide the necessary effect such as inhibiting phosphorylation of kinases or treating disease states in mammals. According to the present invention, a suitable single dose size is a dose that can be prevented or treated by an animal with a disease when administered one or more times for a suitable period of time. Doses may vary depending on the disease being treated. For example, in the treatment of hypersensitivity, a suitable single dose may depend on the nature of the immunogen that causes the hypersensitivity.

An effective administration protocol (that is, administering a therapeutic composition in an effective manner) comprises appropriate dose parameters and routes of administration that result in the prevention or treatment of a disease. Suitable dose parameters and routes of administration can be determined using standard methods in the art for a particular disease. Such methods include, for example, determination of survival rates, side effects (i.e. toxicity) and disease progression or regression. For example, the efficacy of the dose parameters and routes of administration of a therapeutic composition of the present invention can be determined by evaluating the response rates. Such response rates refer to the percentage of patients treated in a population of patients who respond with partial or complete remission.

The pharmaceutical formulations of the compounds of the invention to be used for therapeutic administration must be sterile. Sterility is easily achieved by filtration through sterile membranes such as 0.2 micrometer membranes, or by other conventional methods. The formulations are typically stored in lyophilized form or as an aqueous solution. The pH of the preparations of the invention will typically be about 3-11, more preferably about 5-9 and most preferably about 7-8. It will be understood that the use of certain of the above excipients, supports or stabilizers will result in the formation of cyclic salts of polypeptides. While the preferred route of administration is by injection, other administration methods such as oral, intravenous (bolus and / or infusion), subcutaneous, intramuscular, colonic, rectal, nasal, transdermal or intraperitoneal are also anticipated, using various formulations pharmaceuticals such as suppositories, implanted pellets or small cylinders, aerosols, oral pharmaceutical formulations and topical formulations such as ointments, drops and dermal patches. The compounds of the invention are desirably incorporated into shaped articles such as implants that can employ inert materials such as biodegradable polymers or synthetic silicones, for example Silastic, silicone rubber or other commercially available polymers.

The compounds of the invention can also be administered in the form of liposome distribution systems, such as small unilamellar vesicles, large unilamellar vesicles and multilamellar vesicles. Liposomes can be formed from a variety of lipids, such as cholesterol, stearylamine or phosphatidylcholines.

The compounds of the invention can also be administered through the use of antibodies, antibody fragments, growth factors, hormones or other targeting groups, to which the compound molecules are coupled. The compounds of the invention can also be coupled to suitable polymers as addressable drug carriers. Such polymers may include polyvinylpyrrolidone, pyran copolymer, polyhydroxypropyl methacrylamide-phenol, polyhydroxyethyl-aspartame-phenol or polyethylenoxide-polylysine substituted with palmitoyl residues. In addition, the compounds of the invention can be coupled to a class of biodegradable polymers useful for achieving controlled release of a drug, for example polylactic acid, polyglycolic acid, polylactic and polyglycolic acid copolymers, polypropylctone caprolactone, polyhydroxybutyric acid, polyorthoesters, polyacetals, polydihydropyran, polycyanoacrylates and crosslinked or amphipathic block copolymers of hydrogels. Semi-permeable polymeric polymers and matrices can be formed into shaped articles, such as valves, stents, tubes, prostheses and the like.

Liquid formulations of the therapeutic compound in general are placed in a container that has a sterile access port, for example, an intravenous solution bag or a vial that has a buffer pierceable by a hypodermic injection needle.

Therapeutically effective doses can be determined by in vitro or in vivo methods. For each particular compound of the present invention, individual determinations can be made to determine the optimum dose required. The range of therapeutically effective doses will be influenced by the route of administration, the therapeutic objectives and the condition of the patient. For hypodermic needle injection, it can be assumed that the dose is administered in body fluids. For other routes of administration, the absorption efficiency should be determined individually for each compound by methods well known in pharmacology. According to this, it may be necessary for the therapist to dose the dose and modify the route of administration as required to obtain the optimal therapeutic effect. The determination of effective dose levels, that is, the dose levels necessary to achieve the desired result, will be readily determined by the person skilled in the art. Typically, compound applications begin at low dose levels, increasing the dose levels until the desired effect is achieved.

The compounds and compositions of the invention can be administered orally or parenterally in an effective amount in the dose range of about 0.001 to about 1000 mg / kg, preferably from about 0.01 to about 100 mg / kg. and more preferably from about 0.1 to about 20 mg / kg. Advantageously, the compounds and compositions of the invention can be administered several times a day. Other dosage guidelines may also be useful (for example, single daily dose and / or continuous infusion).

Typically, about 0.5 to about 500 mg of a compound or mixture of compounds of the invention, as the acid form or free base or as a pharmaceutically acceptable salt, is mixed with a carrier, carrier, excipient, binder, preservative , stabilizer, coloring, flavoring, etc., physiologically acceptable, as required by accepted pharmaceutical practice. The amount of active ingredient in these compositions is such that an adequate dose is obtained in the indicated range.

Typical adjuvants that can be incorporated into tablets, capsules and the like are binders such as gum arabic, corn starch or gelatin, and excipients such as microcrystalline cellulose, disintegrating agents such as corn starch or alginic acid, lubricants such as magnesium stearate, sweetening agents such as sucrose or lactose, or flavoring agents. When the pharmaceutical form is a capsule, in addition to the above materials it may also contain liquid carriers such as water, saline or a fatty oil. Other materials of various types can be used as coatings or as modifiers of the physical form of the pharmaceutical form. Sterile injection compositions may be formulated according to conventional pharmaceutical practice. For example, dissolution or suspension of active compound in a vehicle such as an oil or a synthetic fatty vehicle such as ethyl oleate or in a liposome may be desired. Buffers, preservatives, antioxidants and the like may be incorporated according to accepted pharmaceutical practice.

Acid addition salts include hydrochlorides, hydrobromides, iohydrates, alkylsulfonates, for example, methanesulfonates, ethanesulfonates or isethionates, arylsulfonates, for example, p-toluenesulfonates, besylates or napsylates, phosphates, sulfates, hydrogen sulfates, acetates, trifluoroacetates, citrates, propylates maleates, fumarates, malonates, succinates, lactates, oxalates, tartrates and benzoates. Salts derived from inorganic or organic bases include alkali metal salts such as sodium or potassium salts, alkaline earth metal salts such as magnesium or calcium salts, and salts of organic amines such as morpholine, piperidine, dimethylamine or diethylamine salts. Prodrugs of the compounds of formula (1) include those compounds, for example, esters, alcohols or aminos that are convertible in vivo by metabolic means, for example, by hydrolysis, reduction, oxidation or transesterification, to compounds of formula (1) . Particularly useful salts of the compounds according to the invention include pharmaceutically acceptable salts, especially pharmaceutically acceptable acid addition salts. Next, the pharmacological activity of the compounds of the present invention is specifically explained by test examples.

The pharmacological activities of the compounds of the present invention are obtained according to the example test procedures as follows, for example.

Type 1 biological test

Inhibitory effect on compounds on autophosphorylation of platelet-derived growth factor receptor β-PDGF

(1) Phosphorylation assay in HR5

The HR5 cell line is a CHO cell cell line manipulated to overexpress human β-PDGFR, a cell line that is available from the ATCC. The level of β-PDGFR expression in HR5 cells is about 5 x 104 receptors per cell. For the phosphorylation assay according to the invention, HR5 cells were grown to confluence in 96-well microtiter plates under standard tissue culture conditions, followed by serum fasting for 16 hours. The quiescent cells were incubated at 37 ° C without or with increasing concentrations of the test compound (0.01-30 µM) for 30 minutes followed by the addition of 8 nM BB PDGF for 10 minutes. The cells were lysed in 100 mM Tris, pH 7.5, 750 mM NaCl, 0.5% Triton X-100, 10 mM sodium pyrophosphate, 50 mM NaF, 10 ug / ml aprotinin, 10 ug / ml leupeptin, 1 mM phenylmethylsulfonyl fluoride, 1 mM sodium vanadate and the lysate was clarified by centrifugation at 15,000 xg for 5 minutes. The clarified lysates were transferred to a second microtiter plate in which the wells had previously been coated with 500 ng / well of the anti-β-PDGFR 1B5B11 Acm, and then incubated for two hours at room temperature. After washing three times with binding buffer (3% gelatin, 25 mM Hepes, pH 7.5, 100 mM NaCl, 0.01% Tween-20), 250 ng / ml of rabbit anti-rabbit polyclonal antibody was added -phosphotyrosine (Transduction Laboratory) and the plates were incubated at 37 ° C for 60 minutes. Subsequently, each well was washed three times with binding buffer and incubated with 1 ug / ml of horseradish peroxidase-conjugated anti-rabbit antibody (Boehringer Mannheim) at 37 ° C for 60 minutes. The wells were washed before adding ABTS (Sigma), and the formation rate of

5

fifteen

25

35

Four. Five

55

650 nm substrate. The test results are described as IC50 (expressed as the concentration of a compound according to the invention that inhibits phosphorylation of the PDGF receptor by 50%) compared to control cells that are not exposed to a compound according to the invention.

Examples of such IC50 test results in the HR5 test for compounds according to the invention are shown below in Table 1.

(1) Phosphorylation test in MG63

The MG63 cell line is a human osteosarcoma tumor cell line available from the ATCC. This assay is to measure phosphorylation of endogenous β-PDGFR in MG63 cells. The test conditions are the same as those described for HR5 cells, except that stimulation with PBGF-BB is provided in the presence or absence of 45% human plasma. The results of the HR5 assay are described as an IC50 (expressed as the concentration of a compound according to the invention that inhibits phosphorylation of the PDGF receptor by 50%) compared to control cells that are not exposed to a compound according to the invention.

Examples of such IC50 test results in the MG63 test for compounds according to the invention are shown below in Table 1.

The results for the examples of compounds 1 and 2 are shown in table 1 below.

Table 1

Compound example

MG63 with human plasma IC50 (µM) HR5 IC50 (µM)

Example 1

0.030 0.250

Example 2

0.060 0,130

Type 2 biological test

Growth inhibition against smooth muscle cells

Vascular smooth muscle cells are isolated from a pig aorta by explantation and are used for testing. The cells are placed in wells of a 96-well plate (8000 cells / well) and grown in Dulbecco-modified Eagle's medium (DMEM; Nissui Pharmaceutical Co., Ltd.) with 10% fetal bovine serum (SBF; Hyclone ) for 4 days. The cells are then further cultured in DMEM with 0.1% SBF for 3 days and synchronized in a stationary phase of cell growth.

DMEM with 0.1% SBF and a test sample at a varied concentration are added to each well, and cell growth with PDGF-BB (SIGMA, final concentration: 20 ng / ml) is caused. After culturing for three days, cell growth is measured using a cell growth assay kit (Boehringer Mannheim) according to the XTT method [J. Immunol Methods, 142, 257-265 (1991)], and the cell growth score is calculated by the following equation

Cell growth score = 100 X {1- (M-PO) / (P100-PO)} where P100 = absorbance by the XTT reagent when stimulated with PDGF-BB; PO = absorbance by the XTT reagent when not stimulated with PDGF-BB, and M = absorbance by the XTT reagent after the addition of a sample when stimulated with PDGF-BB.

The test result is expressed as the concentration of a test compound that inhibits cell growth by 50% (IC50).

Type 3 biological test

Inhibitory effect on hypertrophy of vascular intima

Male SD rats (weight: 375-445 g, Charles River, reference criterion) are anesthetized with sodium pentobarbital (50 mg / kg, ip), and then the neck of each animal is cut by medium incision, followed by retrograde insertion of a balloon catheter (2F, Edwards Laboratories) in the left outer carotid. After repeating the previous treatment seven times, the catheter is removed, the left external carotid is ligated and the wound is sutured. A test compound is suspended in a 0.5% solution of Tween 80 in an aqueous solution of sodium chloride to a concentration of 20 mg / ml in the case of intraperitoneal administration and in a 0.5% solution of methylcellulose 400 up to a concentration of 6 mg / ml in the case of oral administration. The suspension is administered once a day in the case of intraperitoneal administration and once or twice a day in the case of oral administration for a period of 15 days beginning the day before the balloon injury. On day 14 after the balloon injury, the animal is killed and its left carotid is removed. The tissues are fixed with formalin, wrapped in paraffin and cut, followed by Elastic Wangeeson staining. The cross-sectional area of vascular tissues (intimate and medium) is measured with an image analyzer (Luzex F, NIRECO) and the ratio of the intimate / medium area (I / M) is considered as the degree of hypertrophy of the vascular intima

Of the results obtained, it is apparent when the hypertrophy of the vascular intima is significantly inhibited by the administration of the compounds of the present invention.

Type 4 biological test

Evaluation by the use of an adjuvant arthritis model in rat

Dead Mycobacterium bacterium (Difco Laboratories Inc.) cells are broken in an agate mortar and suspended in liquid paraffin at the final concentration of 6.6 mg / ml, followed by high pressure steam sterilization. Then, 100 ml of the suspension in the right front foot pad of each animal from groups of 8-week-old female Lewis rats (Charles River Japan) (6 animals / group) are injected subcutaneously to induce adjuvant arthritis. A test compound is suspended in a 0.5% solution of methylcellulose at the final concentration of 3 mg / ml and, from just before the induction of arthritis, the suspension is administered orally in an amount of 100 ml. / 100 g body weight once a day, 5 days a week. A 0.5% solution of methylcellulose is administered to a control group. A normal group is not given adjuvant treatment or administration of test compound. Administration of the test compound continues until day 18 after treatment with the adjuvant. On day 17, the number of leukocytes in peripheral blood is counted, and on day 18 all blood is collected, followed by dissection.

The change in body weight is measured and evaluated with the passage of time, the change of edema in the front foot with the passage of time, the weight of spleen and thymus, the number of leukocytes in peripheral blood, the hydroxyproline content of the urine, urine glycosaminoglycan content, serum SH concentration, serum nitrogen monoxide concentration and serum mucoprotein concentration. The volume of each of both front feet is measured using a rat front foot edema measurement device (TK-101, Unicom). The number of leukocytes in peripheral blood is counted using a multichannel automated blood cell counter (Sysmex K-2000, Toa, Iyo Denshi Co., Ltd.). Urine hydroxyproline content is measured according to the method described in Ikeda, et al., Annual Report of Tokyo Metropolitan Research Laboratories PH, 36, 277 (1985), and glycosaminoglycan content is measured according to the method described in Moriyama, et al., Hinyo Kiyo, 40, 565 (1994) and Klompmakers, et al., Analytical Biochemistry, 153, 80 (1986). The serum SH concentration is measured according to the method described in Miesel, et al., Inflammation, 17, 595 (1993), and the nitrogen monoxide concentration is measured according to the method of Tracey, et al., Journal of Pharmacology & Experimental Therapeutics, 272, 1011 (1995). The mucoprotein concentration is measured using the Aspro GP kit (Otsuka Pharmaceutical Co., Ltd.). The percent inhibition is calculated for each indication according to the following equation.

% inhibition = {(Control group -Group with administered compound) / (Control group - Normal group)} x 100.

Of the results obtained from such tests, it is apparent when the compound according to the invention inhibits the appearance of adjuvant arthritis.

Type 5 biological test

Activity in a proliferative mesangial glomerulonephritis model

Anti-Thy-1.1 monoclonal antibody from rat OX-7 (Sedaren) is administered to male Wister-Kyoto rats (Charles River Japan, 160 g, 6 animals / group) in an amount of 1.0 mg / kg by intravenous administration to through the tail vein. A test compound is suspended in a 0.5% methylcellulose solution and the resulting suspension is administered to each of the rats twice daily for a period of 7 days beginning the day before the administration of OX-7. On day 7 after administration of OX-7, when mesangial cell growth and extracellular matrix hypertrophy become important, the left kidney of each rat is removed, fixed with 20% buffered formalin for 6 hours and in paraffin wrap, followed by cuts. The pieces obtained are subjected to tissue immunostaining using the PC10 antibody (DAKO) against an intranuclear proliferative cell antigen. After comparative staining with methyl green staining solution using diaminobenzidine as a color developer, the paraffin pieces are enclosed. Half of the glomeruli are observed in a piece of kidney and the number of cells in a glomerulus that are positive for the intranuclear proliferative cell antigen is calculated. The test for the significance of the difference is carried out by the Wilcox test.

Of such results, it is apparent when the compounds according to the present invention show relieving activity on proliferative mesangial glomerulonephritis.

Claims (11)

1. A process to make a compound according to formula A (2): image 1 Formula A (2) wherein R1 is a member selected from the group consisting of: -CN, -O-C1-8 alkyl having straight or branched chain; -O-phenyl, -O-naphthyl, -O-indolyl and -O-isoquinolyl; n is from 2 to 5; R3 is a member selected from the group consisting of: piperidine, pyrrolidine, morpholine, piperazine, 4-methyl-piperidine and 2-methyl-piperidine; and all pharmaceutically acceptable salts, hydrates and solvates thereof; which comprises the steps of: 15 (a) etherifying the hydroxy group of a compound of formula I with a compound of formula II wherein each of L1 and L2 is independently a leaving group selected from the group consisting of halogen, alkoxy, aryloxy, alkylthio, alkylsulfinyl, alkylsulfonyl, alkylsulfonyloxy and arylsulfonyloxy, wherein L1 is less reactive than the leaving group L2 under basic etherification conditions in the presence of a solvent at temperature 20 refluxing the solvent for 2-6 hours to produce a compound of formula III as follows: image2

(b)

nitrate the compound according to formula III, to give a compound according to formula IV, at a temperature from 0 ° C to 80 ° C in nitric acid and a solvent, as follows:

(C)

reacting the compound of formula IV, with an amine-containing compound, in the presence of a basic catalyst and a solvent to provide a compound of formula V, as follows:

(d)

reduce the nitro group in the compound of formula V to an amino group and thus produce a compound of formula VI, as follows:

(and)

reacting the compound of formula VI with ammonium and formamide format at a temperature from 100 ° C to 200 ° C to produce a cyclic quinazoline derivative of formula VII, as follows:

(F)

replace the hydroxy group of the compound of formula VII with a leaving group Q selected from the group consisting of halogen, alkoxy, aryloxy, alkylthio, alkylsulfinyl, alkylsulfonyl, alkylsulfonyloxy, arylsulfonyloxy to provide a compound of formula VIII, as follows:

(g)

react the compound of formula VIII with a compound of formula IX to replace leaving group Q and provide a compound of formula A (2), as follows:

image2 image3 image2 image4 image5 image 1 15 (h) and optionally, produce a salt of the compound of formula A (2). 2. The process according to claim 1 wherein the leaving group L1 in step (a) is selected from the group consisting of Cl, Br, p-toluenesulfonate and methylsulfonate. The process according to claim 1 wherein the leaving group L2 in step (a) is selected from the group consisting of Cl, Br, p-toluenesulfonate and methylsulfonate. 4. The process according to claim 1 wherein the solvent in step (a) is selected from the group consisting in toluene, methanol, ethanol, ether and THF. 25 5. The process according to claim 1 wherein the base in step (a) is selected from the group consisting of potassium carbonate, sodium carbonate and sodium hydroxide. 6. The process according to claim 1 wherein the catalyst in step (c) is selected from the group consisting of potassium carbonate, sodium carbonate, sodium hydroxide and sodium iodide.

7.

The process according to claim 1 wherein the solvent in step (c) is selected from the group consisting of toluene, ethanol, ether, THF, glyme, diglyme and MBTE.

8.

The process according to claim 1 wherein the leaving group Q is selected from the group consisting of Cl, Br, p-toluenesulfonate and methylsulfonate.

The process according to claim 1, wherein R1 is selected from the group consisting of -O-isopropyl and CN, and n is 2 or 3. 10. The process according to claim 9, wherein R3 is piperidine, methyl-piperidinine, 2-methylpiperidine or pyrrolidine 10 11. The process according to claim 1, wherein n is 3, and R1 is -O-isopropyl or CN. 12. The process according to claim 1, wherein R3 is N-piperidine or N-pyrrolidine. 13. The process according to claim 9, wherein R3 is N-piperidine or N-pyrrolidine. 14. The process according to claim 1, wherein n is 3, R1 is -O-isopropyl, and R3 is N-piperidine or N-morpholine.