HK1146722B - Equilibrative nucleoside transporter ent1 inhibitors - Google Patents

Description

Balanced nucleoside transporter ENT1 inhibitors

The present invention relates to novel compounds of formula (I) having equilibrium nucleoside transporter ENT1 inhibitory properties, pharmaceutical compositions comprising these compounds, chemical processes for the preparation of these compounds and their use in the treatment of diseases associated with the inhibition of ENT1 receptors in animals, particularly humans.

The penetration of nucleosides across cell membranes requires specific transporters. Among the family of nucleoside transporters, the Equilibrium Nucleoside Transporter (ENT) is the most widely expressed and four human ENTs have been identified in humans: hENT-1, hENT-2, hENT-3, and hENT-4. Most thoroughly characterized are hENT-1 and hENT-2, which are cell surface proteins and are broadly selective for purine and pyrimidine nucleosides. Using their sensitivity to inhibition via nitrobenzyl mercaptopurine Nucleosides (NBMPR) can be distinguished from each other. ENT1 can be effectively inhibited via nanomolar NBMPR and is therefore also referred to as NBMPR sensitive equilibrium nucleoside transporter. ENT2 is not sensitive to nanomolar concentrations of NBMPR, but can be inhibited via higher concentrations (micromolar concentrations) of NBMPR and is therefore also referred to as NBMPR-Insensitive Equilibrium Nucleoside Transporter (iENTP) [ see Griffith et al, biochim. 153-181(1986)].

Adenosine is an endogenous purine nucleoside which is released in particular in pathophysiological conditions such as ischemia, inflammation and pain. Under these circumstances, it plays an important role in neural and immune regulation. Adenosine administration is used to relieve pain in a variety of forms that cause injury in humans. Because of the short half-life of adenosine and the side effects caused by its administration, there is considerable interest in finding ways to potentiate the effects of endogenous adenosine. The inhibitory effect of ENT1 prevents the uptake of adenosine into cells and may enhance its effect.

Disclosure of Invention

The invention relates to compounds of formula (I)

Including any stereochemically isomeric form thereof, wherein

-A-B-represents

Wherein n is an integer of 0 or 1;

R¹³represents hydroxy or halogen;

R¹⁴represents hydrogen or C1-6 alkyl;

in the divalent groups (a-4), (a-5) and (a-6), any hydrogen atom on the same or different carbon atom may be replaced by a halogen;

R¹and R²Each independently selected from hydrogen, halogen or C_1-6An alkyl group;

R³、R⁴、R⁵and R⁶Each independently selected from hydrogen, halogen, C_1-6Alkyl, polyhalo C_1-6Alkyl radical, C_3-6Cycloalkyl, NO₂Ring of¹Ring of²Or X-R⁸Wherein X represents O or NR⁹，

Wherein R is⁹Is hydrogen, C_1-6Alkyl or C_1-6Alkoxy radical C_1-6Alkyl radical, and

wherein R is⁸Is hydrogen, C_1-6Alkyl radical, C_2-6Alkenyl radical, C_3-6Cycloalkyl, polyhaloC_1-6Alkyl radical, C_1-6Alkoxycarbonyl, C_1-6Alkyl carbonylPolyhalo C_1-6Alkyl carbonyl, ring²、-(C＝O)-(CH₂)_m-Ring²、-(C＝O)-(CH₂)_m-CH₂-OH、-(C＝O)-(CH₂)_m-CH₂-O-C_1-4Alkyl or

Via halogen, hydroxy, cyano, C_3-6Cycloalkyl radical, C_1-6Alkoxy, aminocarbonyl, phenyl, cyclic¹Or a ring²Substituted C_1-6Alkyl, or NR¹¹R¹²Wherein R is¹¹And R¹²Each independently selected from hydrogen and C_1-6Alkyl radical, C_1-6Alkylcarbonyl, or C_1-6An alkoxycarbonyl group;

m is an integer 0, 1 or 2;

ring (C)¹Is selected from

Wherein R is¹⁰Is hydrogen, C_1-6Alkyl radical, C_1-6Alkylcarbonyl, or C_1-6An alkoxycarbonyl group; and is

Ring (C)²Is selected from

Wherein R is¹⁰Is hydrogen, C_1-6Alkyl radical, C_1-6Alkylcarbonyl group, C_1-6Alkoxycarbonyl or halogen-or hydroxy-substituted C_1-6An alkoxycarbonyl group;

or a pharmaceutically acceptable acid addition salt thereof, or a solvate thereof, or an N-oxide form thereof.

When used in the above definition of the terms,

halogen means in general fluorine, chlorine, bromine and iodine radicals;

-C_1-4alkyl is defined as straight and branched chain saturated hydrocarbon groups containing 1 to 4 carbon atoms, such as methyl, ethyl, propyl, butyl, 1-methylethyl, 2-methylpropyl and the like;

-C_1-6alkyl means containing C_1-4Alkyl and higher homologues containing 5 or 6 carbon atoms (homologues), such as 2-methylbutyl, pentyl, hexyl and the like;

-C_3-6alkenyl is defined as straight and branched chain unsaturated hydrocarbon radicals containing from 3 to 6 carbon atoms, such as propenyl, butenyl, pentenyl or hexenyl;

-polyhalo C_1-4Alkyl is defined as polyhalo-substituted C_1-4Alkyl, especially C, substituted by 2 to 6 halogen atoms_1-4Alkyl (according to the above definition), such as difluoromethyl, trifluoromethyl, trifluoroethyl, and the like;

-polyhalo C_1-6Alkyl is defined as polyhalo-substituted C_1-6Alkyl, especially C, substituted by 2 to 6 halogen atoms_1-6Alkyl (according to the above definition), such as difluoromethyl, trifluoromethyl, trifluoroethyl, and the like;

-C_3-6cycloalkyl is generally referred to as cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl.

The term "stereochemically isomeric forms" when used hereinbefore defines all the possible isomeric forms which the compounds of formula (I) possess. Unless otherwise stated or indicated, the chemical designation of a compound refers to the mixture of all possible stereochemically isomeric forms, said mixtures containing all diastereomers and enantiomers of the basic molecular structure. More particularly, the stereoisomeric centers may have the R-or S-configuration; the substituents on the divalent cyclic (partially) saturated radicals may have cis-or trans-configuration. When the substituent R is¹And R²In different sense, the bond between the aryl and carbonyl forms the axis of chirality. With an axisEnantiomers of chiral compounds generally provide stereochemical designation R_aAnd S_a(or aR or aS) and such enantiomers are also included within the term "stereochemically isomeric forms".

Stereochemically isomeric forms of the compounds of formula (I) are expressly included within the scope of the present invention.

The absolute stereochemical configuration of the compounds of formula (I) and the intermediates used in their preparation can be readily determined by those skilled in the art using well-known methods, such as X-ray diffraction.

Furthermore, some compounds of formula (I) and some intermediates used in their preparation may exhibit polymorphism (polymorphism). It is to be understood that the present invention encompasses polymorphic forms having properties useful in addressing the above-mentioned situations.

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

Conversely, the salt form may be converted to the free base form by treatment with a suitable base.

The compounds of formula (I) may exist in unsolvated as well as solvated forms. The term "solvate" is used herein to describe a molecular association comprising a compound of the invention and one or more pharmaceutically acceptable solvent molecules, such as water or ethanol. The term "hydrate" is used when the solvent is water.

The N-oxide forms of the compounds of formula (I), which may be prepared in a manner known in the art, refer to those comprising oxidation of one or several nitrogen atoms in the compounds of formula (I) to the N-oxide. Particularly contemplated are those N-oxides in which the piperidine-nitrogen is N-oxidized.

Interesting compounds of formula (I) are those wherein one or more of the following limitations apply:

a)R¹and R²Are all halogen, especially chloro; or

b)R¹And R²Are all C_1-4Alkyl, especially methyl; or

c) The group A-B-represents (a-1); or

d) The group A-B-represents (a-2); or

e)R³、R⁴、R⁵And R⁶Each independently selected from hydrogen, halogen, C_1-6Alkyl, or X-R⁸Wherein X represents O; or

f)R³、R⁴、R⁵And R⁶Each independently selected from hydrogen, halogen, C_1-6Alkyl, or X-R⁸Wherein X represents O and R⁸Represents C_1-6Alkyl or Het²(ii) a Or

g)R³、R⁴、R⁵And R⁶Each independently selected from hydrogen, halogen, C_1-6Alkyl, or X-R⁸Wherein X represents O and R⁸Represents Het²Wherein Het is²Represents a group (c-2); or

h)R³、R⁴、R⁵And R⁶Each independently selected from hydrogen, halogen, C_1-6Alkyl, or X-R⁸Wherein X represents NR⁹(ii) a Or

i)R³、R⁴、R⁵And R⁶Each independently selected from hydrogen, halogen, C_1-6Alkyl, or X-R⁸Wherein R is⁸Is hydrogen and X represents NR⁹Wherein R is⁹Represents hydrogen; or

j)R³Is hydrogen; or

k)R⁴Is hydrogen, C_1-6Alkyl or halogen; or

l)R⁵Is X-R⁸(ii) a Or

m)R⁶Is halogen; or

n) n is an integer 0.

Other interesting compounds are N- (2- {7- [ (1-acetylpiperidin-4-yl) oxy ] -6-chloro-4-oxo-3, 4-dihydro-1 'H-spiro [ chromene-2, 4' -piperidin ] -1 '-yl } ethyl) -2, 6-dichlorobenzylamide or N- [2- (7-amino-8-chloro-4-oxo-3, 4-dihydro-1' H-spiro [1, 3-benzoxazine-2, 4 '-piperidin ] -1' -yl } ethyl) -2, 6-dichlorobenzylamide, or a pharmaceutically acceptable acid addition salt thereof.

Different methods for the preparation of the compounds of formula (I) are described in the following paragraphs. To simplify the structural formulae of the compounds of formula (I) and intermediates in their preparation, the benzamide moiety hereinafter will be represented by the symbol T.

Compounds of formula (I) may generally be prepared by N-alkylating an intermediate of formula (III) with an intermediate of formula (II) wherein W is a suitable leaving group such as a halogen (e.g., fluorine, chlorine, bromine, iodine) or in some cases W may also be a reactive leaving group such as a sulfonyloxy group such as a methanesulfonyloxy group, a benzenesulfonyloxy group, a trifluoromethylsulfonyloxy group, and the like. This reaction can be carried out in a reaction-inert solvent such as acetonitrile, dichloromethane or dimethylformamide, and optionally in the presence of a suitable base such as sodium carbonate, potassium carbonate, triethylamine or Diisopropylethylamine (DIPEA). Agitation may facilitate the reaction rate. The reaction is preferably carried out at a temperature of about 0 ℃.

The compounds of formula (I) may also be prepared by reacting an intermediate of formula (IV) with an intermediate of formula (V). This reaction can be carried out in a reaction-inert solvent such as dichloromethane or dimethylformamide, and optionally in the presence of a suitable base such as Diisopropylethylamine (DIPEA).

The compounds of formula (I-a) are defined as compounds of formula (I) wherein n is 0 wherein-A-B-represents a group (a-1) which can be prepared by reacting an intermediate of formula (VI) with an intermediate of formula (VII) in a reaction-inert solvent such as methanol in the presence of pyrrolidine.

The compounds of formula (I-B) are defined as compounds of formula (I) wherein n is 0 and wherein-A-B-represents a group (a-2) which can be prepared by reacting an intermediate of formula (VIII) with an intermediate of formula (IX) in a reaction-inert solvent such as toluene in the presence of pyrrolidine or toluene in the presence of p-toluenesulfonic acid and molecular sieves.

The compounds of formula (I-c) are defined as compounds of formula (I) wherein n is 0 and wherein A-B-represents a group (a-4) which can be converted to compounds of formula (I-a) by catalytic hydrogenation conditions, for example by using hydrogen and a catalyst such as Raney nickel in a reaction-inert solvent such as methanol.

The compound of formula (I-a) may be converted to a compound of formula (I-d-1) by reduction methods known in the art (e.g. treatment with sodium borohydride in a suitable solvent such as methanol), the compound of formula (I-d-1) being defined as the group (a-6) represented by-A-B-in the compound of formula (I), wherein R is¹¹Represents a hydroxyl group and n is 0.

The compound of formula (I-d-1) can be converted to the compound of formula (I-c) by treatment with trimethylsilylhydride in a reaction-inert solvent such as dichloromethane in the presence of trifluoroacetic acid.

The compound of formula (I-a) can be converted to a compound of formula (I-e) defined as-A-B-representing group (a-3) in the compound of formula (I) wherein n is 0, by treatment with hydroxylamine under basic conditions.

The compound of formula (I-d-1) can be converted into the compound of formula (I-f), which is defined as the group (a-5) represented by-A-B-in the compound of formula (I), wherein n is 0, by treatment with hydrochloric acid in a reaction-inert solvent such as THF.

The compounds of formula (I-f) may be converted to compounds of formula (I-C) using catalytic hydrogenation conditions, for example by using hydrogen and a catalyst such as Pd/C in a reaction inert solvent such as methanol.

The compounds of formula (I) may also be converted to the corresponding N-oxide form according to methods known in the art for converting trivalent nitrogen to its N-oxide form. The N-oxidation reaction is generally carried out by reacting the starting material of formula (I) with a suitable organic or inorganic peroxide. Suitable inorganic peroxides include, for example, hydrogen peroxide, alkali or alkaline earth metal peroxides (e.g., sodium peroxide, potassium peroxide); suitable organic peroxides may include peroxyacids such as benzoates (benzoates) or halogen-substituted peroxybenzoates such as 3-chloroperoxybenzoic acid, peroxyalkanoic acids such as peracetic acid, alkyl hydroperoxides such as t-butyl hydroperoxide. Suitable solvents are, for example, water, lower alkanols, such as ethanol and the like, hydrocarbons, such as toluene, ketones, such as 2-butanone, halogenated hydrocarbons, such as methylene chloride, and mixtures of these solvents.

The starting materials and some intermediates are known compounds and are commercially available or can be prepared according to conventional reaction methods generally known in the art. The compounds of formula (I), prepared according to the above-described processes, can be synthesized as racemic mixtures of enantiomers, which can be separated from each other according to resolution methods known in the art. These compounds of formula (I), obtained in racemic form, can be converted into the corresponding diastereomeric salt forms by reaction with a suitable chiral acid. The salt forms of the diastereomers are then separated, for example, by selective or fractional crystallization and the enantiomers are liberated therefrom by base. An alternative way of separating the enantiomeric forms of the compounds of formula (I) involves liquid chromatography using a chiral stationary phase. The pure stereochemically isomeric forms may also be obtained from the corresponding pure stereochemically isomeric forms of the appropriate starting materials, provided that the reaction proceeds stereospecifically. If a particular stereoisomer is desired, it is preferred that the compound be synthesized by stereospecific methods of preparation. These processes will advantageously use enantiomerically pure starting materials.

The compounds of formula (I), their pharmaceutically acceptable salts and stereoisomeric forms, have the property of inhibiting the balanced nucleoside transporter ENT1, as demonstrated according to pharmacological example c.1.

Accordingly, the compounds of formula (I) of the present invention may be used as medicaments, in particular for the treatment of conditions or diseases mediated by the inhibitory activity of the balanced nucleoside transporter ENT1, in particular the balanced nucleoside transporter ENT 1. Subsequently, the compounds of the invention can be used for the manufacture of a medicament for the treatment of a condition or disease mediated by the activity of the equilibrative nucleoside transporter ENT1, in particular the inhibitory activity of the equilibrative nucleoside transporter ENT 1.

The invention also provides the use of a compound of formula (I) or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for the treatment of a condition or disease selected from the group consisting of a condition or disease of the balanced nucleoside transporter ENT 1.

In a particular embodiment, the present invention provides a compound of formula (I) for use as a medicament or for the treatment of a condition or disease selected from the group consisting of a condition or disease of the balanced nucleoside transporter ENT 1.

Furthermore, the present invention provides a method for treating a condition mediated by the activity of the balanced nucleoside transporter ENT1 in a mammalian subject, which comprises administering to a mammal in need of such treatment a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof.

Symptoms or diseases mediated by the balanced nucleoside transporter ENT1 are, for example, acute and chronic pain symptoms, including inflammatory pain, neuropathic pain, cancer pain, cardioprotection, cerebral protection, Traumatic Brain Injury (TBI), spinal cord protection, neuroprotection, chronic pressure skin ulcers, wound healing, anti-spasticity, organ transplantation (organ preservation, e.g. cardioplegia), sleep disorders, pancreatitis, glomerulonephritis, and anti-thrombosis (anti-platelet).

Chronic pain symptoms are associated with hyperalgesia and allodynia (allodynia). These symptoms may include acute pain, skeletal muscle pain, lower back pain and radiculopathy, upper limb pain, fibromyalgia and myofascial pain syndromes, oromandibular facial pain, abdominal pain, phantom pain, trigeminal and atypical facial pain, radiculoinjury and arachnoiditis, geriatric pain, central neuralgia, inflammatory pain.

Neurodegenerative pain results from lesions of the peripheral or central nervous system. It is usually associated with somatosensory deficits and the distribution of pain is mostly associated with areas of somatosensory dysfunction. The onset of pain can be delayed after the causative event, even up to months or years. Neurodegenerative pain has many causes and has many different symptoms and neurological deficits. Examples are peripheral nerve injuries resulting from traumatic injury compression, ischemia, toxins, nutritional deficiencies, viral infections of the liver and kidney and complications.

The terms "treating" and "treatment," as used herein, refer to therapeutic, palliative, and prophylactic treatment, including reversing, alleviating, inhibiting progression, or preventing a disease, disorder, or condition for which the term is applicable, or one or more symptoms of such disease, disorder, or condition.

In addition, the present invention provides pharmaceutical compositions comprising at least one pharmaceutically acceptable carrier and a therapeutically effective amount of a compound of formula (I).

To prepare the pharmaceutical compositions of this invention, an effective amount of the particular compound, in base or acid addition salt form, as the active ingredient is combined in intimate admixture with at least one pharmaceutically acceptable carrier, which carrier may take a wide variety of forms depending on the form of preparation desired for administration. These pharmaceutical compositions are suitably in unit dosage form, preferably for oral, rectal, subcutaneous or parenteral injection.

For example, in the preparation of compositions in the form for oral administration, e.g., in the case of oral liquid preparations such as suspensions, syrups, elixirs and solutions, any of the usual liquid pharmaceutical carriers may be employed such as water, glycols, oils, alcohols and the like; or in the case of powders, pills, capsules and tablets, solid pharmaceutical carriers such as starches, sugars, kaolin, lubricants, binders, disintegrating agents and the like may be employed. Because of their ease of administration, tablets and capsules represent the most advantageous oral unit dosage form in which case solid pharmaceutical carriers are obviously employed. For parenteral injection compositions, the pharmaceutical carrier consists essentially of sterile water, however, other ingredients may be included to improve the solubility of the active ingredient. Injectable solutions can be prepared, for example, by using a pharmaceutical carrier containing saline solution, dextrose solution, or a mixture of the two. Injectable suspensions may also be prepared by using suitable liquid carriers, suspending agents and the like. In compositions suitable for subcutaneous administration, the pharmaceutical carrier may optionally contain penetration enhancers and/or suitable wetting agents, optionally in combination with suitable additives in minor proportions that do not cause a significant deleterious effect on the skin. The additives may be selected to facilitate administration of the active ingredient to the skin and/or to aid in the preparation of the desired composition. These topical compositions can be administered in different ways, for example as transdermal patches, patches (spot-on) or ointments. Addition salts of the compounds of formula (I) are significantly more suitable for the preparation of aqueous compositions because of their increased aqueous solubility relative to the corresponding base forms.

It is particularly advantageous to formulate the pharmaceutical compositions of the present invention in unit dosage form for convenient administration and uniform dosage. "Unit dosage form" as used herein, refers to physically discrete units suitable as unitary dosages, each unit containing a predetermined quantity of active ingredient calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier. Examples of such unit dosage forms are tablets (including scored or coated tablets), capsules, pills, powder packets, glutinous rice, injectable solutions or suspensions, teaspoonful, tablespoonful and the like, and isolated multiples thereof.

For oral administration, the pharmaceutical compositions of the present invention may take the form of solid dosage forms, such as tablets (swallowable and chewable forms), capsules or caplets (gelcaps) prepared by conventional methods using pharmaceutically acceptable excipients and carriers such as binding agents (e.g., pregelatinized corn starch, povidone, hydroxypropylmethyl cellulose, and the like), fillers (e.g., lactose, microcrystalline cellulose, calcium phosphate, and the like), lubricants (e.g., magnesium stearate, talc, silicon dioxide, and the like), disintegrants (e.g., potato starch, sodium starch glycolate, and the like), wetting agents (e.g., sodium lauryl sulfate), and the like. The tablets may also be coated by methods well known in the art.

Liquid preparations for oral administration may take the form of, for example, solutions, syrups or suspensions, or they may be formulated as a dry product for constitution with water and/or other suitable liquid carrier before use. Such liquid preparations may be prepared by conventional methods, optionally using other pharmaceutically acceptable additives such as suspending agents (e.g. sorbitol syrup, methyl cellulose, hydroxypropylmethyl cellulose or hydrogenated edible fats), emulsifying agents (e.g. lecithin or acacia), non-aqueous carriers (e.g. almond oil, oily esters or ethanol), sweetening agents, flavouring agents, masking agents and preservatives (e.g. methyl or propyl p-hydroxybenzoate or sorbic acid).

Pharmaceutically acceptable sweeteners which may be used in the pharmaceutical compositions of the present invention preferably include at least one intense sweetener such as aspartame, acesulfame potassium, sodium cyclamate (sodium cyclamate), alitame (alitame), dihydrochalcone sweetener, monellin (monellin), stevioside sucralose (4, 1 ', 6' -trichloro-4, 1 ', 6' -trideoxygalactosucrose), or preferably saccharin, sodium saccharin or calcium, and optionally at least one bulk (bulk) sweetener such as sorbitol, mannitol, fructose, sucrose, maltose, isomalt (isomalt), glucose, hydrogenated glucose syrup, xylitol, caramel or honey. Intense sweeteners are conveniently used in low concentrations. For example, in the case of sodium saccharin, the concentration ranges from about 0.04% to 0.1% (weight/volume) of the final formulation. Bulk sweeteners can be effectively used in relatively high concentrations ranging from about 10% to about 35%, preferably from about 10% to 15% (weight/volume).

The pharmaceutically acceptable flavoring agent which can mask the bitter tasting ingredient in the low dose formulation is preferably a fruity flavor such as cherry, raspberry, blackcurrant or strawberry flavor. The combination of two flavours may give very good results. In high dose formulations, more intense pharmaceutically acceptable flavours such as caramel chocolate, mint, Fantasy etc. may be required. The various flavoring agents are present in the final composition at a concentration ranging from about 0.05% to 1% (weight/volume). Combinations of said strong flavourings are advantageously used. It is preferred that the flavoring agent be used without any change or loss in taste and/or color during the formulation process.

The compounds of formula (I) may be formulated for parenteral administration by injection, conveniently intravenous, intramuscular or subcutaneous injection, for example by bolus injection or continuous intravenous infusion. Formulations for injection may be presented in unit dosage form, e.g., in ampoules or in multi-dose containers, with an added preservative. They may be suspensions, solutions or emulsions in oily or aqueous media, and may contain formulatory agents such as isotonic agents, suspending agents, stabilizing agents and/or dispersing agents. Alternatively, the active ingredients may be presented in powder form for constitution with a suitable vehicle, e.g., sterile, pyrogen-free water, before use.

The compounds of formula (I) may also be formulated in rectal compositions such as suppositories or retention enemas, e.g., containing conventional suppository bases such as cocoa butter and/or other glycerides.

A therapeutically effective amount of a compound of formula (I) will be readily determined from the test results presented herein by those skilled in the art of treating diseases associated with mediation of cannabinoid (cannabinoid) receptors. A therapeutically effective amount is generally considered to be from about 0.001 mg/kg to about 50 mg/kg body weight, more preferably from about 0.01 mg/kg to about 10 mg/kg body weight of the patient to be treated. It may be appropriate to administer the therapeutically effective amounts of two or more sub-dosage forms at appropriate intervals throughout the day. The sub-doses may be formulated in unit dosage forms, for example, containing from about 0.1 mg to about 1000 mg, more particularly from about 1 to about 500 mg, of active ingredient per unit dosage form.

As used herein, a "therapeutically effective amount" of a compound is an amount of the compound that, when administered to an individual or animal, produces a sufficient amount of the compound in the individual or animal to cause a recognizable inhibition of the ENT1 transporter.

The exact dose and frequency of administration will depend on the particular compound of formula (I) used, the particular condition being treated, the severity of the condition being treated, the age, weight and general physical condition of the particular patient and other drugs the patient may be taking and will be well known to those skilled in the art. Moreover, the "therapeutically effective amount" may be decreased or increased depending on the response of the patient being treated and/or depending on the evaluation of the physician prescribing the compounds of the instant invention. The effective daily amounts mentioned above are therefore only guidelines.

Experimental part

The following abbreviations are used in the methods described below: ' CH₃OH "means methanol," CH₂Cl₂"means methylene chloride" CH₃CN ' refers to acetonitrile, ' DIPE 'Refers to diisopropyl ether, "DIPEA" refers to diisopropylethylamine, "MgSO₄"means magnesium sulfate" and "Na₂SO₄"refers to disodium sulfate," Na₂CO₃"refers to disodium carbonate," THF "refers to tetrahydrofuran," EtOH "refers to ethanol," DMF "refers to N, N-dimethylformamide," CF₃COOH "means trifluoroacetic acid," H₂SO₄"refers to sulfuric acid," KOAc "refers to potassium acetate," NH₃"refers to ammonia," NaBH₄"means sodium borohydride" NH₄Cl "refers to ammonium chloride," NaOH "refers to sodium hydroxide," NaBH (OAc)₃"refers to sodium triacetoxyborohydride (sodium triacetoxyborohydride)," Pd (OAc)₂"refers to palladium acetate," BINAP "refers to" 1, 1 '- [1, 1' -binaphthyl]-2, 2' -diylbis [1, 1-diphenylphosphine]”，“Cs₂CO₃"means cesium carbonate" BBr₃"means tribromoborane" BF₃"means trifluoroborane" K₂CO₃"means potassium carbonate" Et₃N 'denotes triethylamine and' NH₂OH "refers to hydroxylamine," NaHCO₃"refers to the monosodium carbonate salt," NaOAc "refers to sodium acetate," Et₂O "means diethyl ether," PTSA "means p-toluenesulfonic acid," DMS "means dimethylsulfide," LiOH "means lithium hydroxide," HCl "means hydrochloric acid," Pd₂(dba)₃"refers to tris [ mu- [ (1, 2-eta: 4, 5-eta) - (1E, 4E) -1, 5-diphenyl-1, 4-pentadien-3-one]]Dipalladium, "HBTU" refers to 1- [ bis (dimethylamino) methylene]-1H-benzotriazolium hexafluorophosphate (1-) 3-oxide, "NH₄HCO₃"refers to monoammonium carbonate," CHC₁₃"refers to trichloromethane," HNO₃"means nitric acid" CH₃NH₂"refers to methylamine and" NH₄OH "refers to ammonium hydroxide," DMSO "refers to dimethyl sulfoxide, and" NaBH₃CN "refers to sodium cyanoborohydride.

The high performance liquid chromatography purification method comprises the following steps:

-purification method a:

the product was purified by reverse phase high performance liquid chromatography (Shandon)C18 bds (base Deactivated silica)8 μm, 250 g, i.d.5 cm). A gradient of three mobile phases (phase A: 0.25% NH in water) was used₄HCO₃A solution; phase B: CH (CH)₃OH; phase C: CH (CH)₃CN). The desired fractions were collected and worked up.

A. Synthesis of intermediates

Example A.1

Preparation ofIntermediate (1)

4 ' -carbamoyl-2 ' -chloro-5 ' -hydroxy-acetanilide (0.081 mol) and H in methanol (150 ml)₂SO₄(15 ml) stirred and refluxed for 1 hour. The methanol was evaporated (vacuum). Water (100 ml) was added. The precipitate was filtered off and dried (vacuum) yielding 15.0 g of intermediate (1).

Example A.2

a) Preparation ofIntermediate (2)

1, 1-Dimethylethyl ester of 4-oxo-1-piperidinecarboxylic acid (0.067 mol), intermediate (1) (0.08 mol) and pyrrolidine (0.0066 mol) in toluene (200 ml) were stirred and refluxed overnight. The toluene was evaporated. The precipitate was filtered off, washed 3 times with DIPE (200 ml) and dried to yield 23.3 g of intermediate (2).

b) Preparation ofIntermediate (3)

CF is prepared by₃COOH/CH₂Cl₂(100 ml; 50/50) slowlyTo intermediate (1) (0.0407 mol) in CH₂Cl₂(50 ml) was stirred in the mixture for 3 hours. The solvent was evaporated. Removing the residue from CH₃CN crystallizes, the precipitate is filtered off and dried, yielding 11.84 g of intermediate (3).

c) Preparation ofIntermediate (4)

CH₃Intermediate (3) (0.031 moles), 2-chloroacetonitrile (0.046 moles) and DIPEA (0.155 moles) were stirred and refluxed in CN (150 ml) for 1 hour. Will CH₃CN evaporated (vacuum). The residue was stirred in water (150 ml) and the precipitate was filtered off, washed with DIPE and dried (vacuum) to yield 9.3 g of intermediate (4).

d) Preparation ofIntermediate (5)

Intermediate (4) (0.03 mol) in NH₃/CH₃The mixture in OH (250 ml) was hydrogenated at 14 ℃ with Raney nickel (catalytic amount) as catalyst in the presence of thiophene solution (1 ml). After absorption (uptake) of hydrogen (2 eq; 1500 ml of hydrogen), the catalyst is filtered off and the filtrate is evaporated. Removing the residue from CH₃CN recrystallizes and the precipitate is filtered off, yielding 6.6 g of intermediate (5).

Example A.3

a) Preparation ofIntermediate (6)

1, 4-dioxa-8-azaspiro [4.5 ] in toluene (4200 ml)]Decane (2.1 moles) was stirred at room temperature. D1PEA (2.3 moles) was added to the reaction mixture. 2-chloro-acetonitrile (2.2 mol) was slowly added to the reaction mixture and then stirred at 80 ℃ for 2 hours. The reaction mixture was allowed to cool. Then mixing the reactionThe material was washed twice with water (2000 ml). The solvent of the separated organic layer was evaporated to give 175.3 g of residue. The 2 separated aqueous layers were combined and treated with CH₂Cl₂(3000 ml) wash. The separated organic layer was dried (Na)₂SO₄) Filtration and evaporation of the solvent (and co-evaporation with toluene) gave another 100 g of residue. The residues were combined and dissolved in CH₃OH/NH₃And then hydrogenated with raney nickel as a catalyst. After uptake of hydrogen (2 equivalents), the mixture was filtered off and the filtrate was evaporated, yielding 242.1 g of intermediate (6).

b) Preparation ofIntermediate (7)

Intermediate (6) (1.04 mol) was added to DIPEA (5.23 mol) and CH₂Cl₂The mixture in (2000 ml) was stirred at 0 ℃.2, 6-Dichlorobenzoyl chloride (1.25 moles) was slowly added dropwise to the reaction mixture at 0 ℃ over 50 minutes. The reaction mixture was allowed to warm to room temperature. Water (2000 ml) was added to the reaction mixture. The separated organic layer was dried (Na)₂SO₄) Filtered and the solvent evaporated. The residue was dissolved in DIPE (1000 ml). The precipitate was filtered off and dried (vacuum) yielding 321 g of intermediate (7).

c) Preparation ofIntermediate (8)

Intermediate (7) (0.180 mol) was dissolved in water (861 ml) and HCl (430 ml, 12M). The reaction mixture was stirred at 80 ℃ for 3 hours. The reaction mixture was poured into 1 kg of ice and Na was added₂CO₃Alkalizing. Adding CH to the aqueous mixture₂Cl₂And (4) extracting. The combined organic layers were dried, filtered and the solvent was evaporated. The residue was stirred in DIPE for 2 hours, then filtered and dried in vacuo to give intermediate (8).

Example A.4

a) Preparation ofIntermediate (9)

A mixture of 4-amino-5-chloro-2-hydroxy-benzylamide (8 g, 0.043 mol) in methanol (250 ml) was hydrogenated with Pd/C (10%) (1 g) as a catalyst in the presence of KOAc (5 g). After uptake of hydrogen (1212 ml), the reaction mixture was used as such with 4-amino-2-hydroxybenzamide (0.0480 mol) and 4-amino-2-hydroxybenzamide (0.048 mol) and the reaction mixture was further hydrogenated at 75 ℃ (100 atm hydrogen pressure) in the presence of thiophene solution (1 ml). After absorption of hydrogen, the catalyst was filtered off and the filtrate was evaporated. The residue was triturated in DIPE, filtered and dried to yield 4.8 g of intermediate (9).

b) Preparation ofIntermediate (10)

50: 50 mixture of regioisomers

Intermediate (9) (0.0051 mol) and 1-chloro-2, 5-pyrrolidinedione (0.0051 mol) in CH₃The mixture in CN was stirred at 50 ℃ for 3 hours. The solvent was evaporated under reduced pressure. Water (50 ml) was added and the resulting white precipitate was filtered off and dried (vacuum, 50 ℃ C., 16 h) to yield 0.850 g of intermediate (10) (a 1: 1 mixture of the two regioisomers).

Example A.5

Preparation ofIntermediate (11)

A mixture of 4-amino-2-hydroxybenzamide (0.039 mol) and cyclopentanone (0.039 mol) in methanol (100 ml) and THF (50 ml) was hydrogenated at 50 ℃ with Pd/C (10%) (1 g) as catalyst in the presence of a thiophene solution (0.5 ml). After absorption of hydrogen (1 eq), the catalyst was filtered off and the filtrate was evaporated. The residue was triturated in DIPE, filtered off and dried to yield 4.1 g of intermediate (11).

Intermediate (12) was prepared according to the same procedure substituting formaldehyde for cyclopentanone.

Intermediate (12)

Example A.6

Preparation ofIntermediate (13)

4 ' -acetyl-2 ' -chloro-5 ' -hydroxy-acetanilide (0.014 mol) was added to H₂SO₄The mixture in (5 ml) and methanol (100 ml) was refluxed for 1 hour. The methanol was evaporated. The reaction mixture was neutralized with aqueous NaOH 1N to pH 7. The mixture is treated with CH₂Cl₂(100 ml) extraction. The separated organic layer was washed with water (100 ml) and brine (100 ml). The separated organic layer was dried (MgSO)₄) Filtration and evaporation of the solvent (vacuum) gave 1.6 g of intermediate (13).

Example A.7

Preparation ofIntermediate (14)

A mixture of 4-amino-2-hydroxybenzamide (0.0328 mol) and benzaldehyde (0.0328 mol) in methanol (100 ml) and THF (50 ml) was prehydrogenated at 50 ℃ with Pd/C (10%) (1 g) as catalyst in the presence of a thiophene solution (0.5 ml). After uptake of hydrogen (1 equivalent), the reaction mixture was allowed to react further. Formaldehyde (1.5 g) was added to the crude reaction mixture and the whole was further hydrogenated at 75 ℃ (under 100 atmospheres of hydrogen). After absorption of hydrogen (1 eq), the catalyst was filtered off and the filtrate was evaporated. The residue was reductively debenzylated by hydrogenation in methanol (150 ml) with Pd/C (10%) (1 g) as catalyst. After uptake of hydrogen (1 eq), the catalyst was filtered off and the filtrate was evaporated, yielding 3.7 g of intermediate (14).

Example A.8

Preparation ofIntermediate (15)

Intermediate (11) (0.0122 mol) was dissolved in DMF (20 ml). 1-chloro-2, 5-pyrrolidinedione (0.0122 mol) was added in one portion. The reaction mixture was stirred at 40 ℃ for 16 hours. The solvent was evaporated under reduced pressure. Water was added to the residue. The precipitate was filtered off and dissolved in ethyl acetate. The organic solution was dried (MgSO₄) Filtered and the solvent evaporated. The residue was purified by column chromatography on silica gel (eluent: ethyl acetate/hexane 50/50 v/v). The product fractions were collected and the solvent was evaporated, yielding 2.5 g of intermediate (15).

Intermediate (16) was prepared according to the same method but starting from intermediate (12).

Intermediate (16)

Example A.9

Preparation ofIntermediate (17)

NH dissolving 5-chloro-2-hydroxy-4- (1-methylethoxy) benzoic acid methyl ester (0.0273 mol) in dioxane₃(0.5N) (300 ml) and the resulting reaction mixture was stirred at 100 deg.C (autoclave) overnight. The solvent was evaporated. The residue was dissolved in 7N NH₃/CH₃OH and the solution was stirred in an autoclave at 100 ℃ overnight. The solvent is evaporated and the residue obtained is subjected to column chromatography on silica gel (eluent: CH)₂Cl₂/CH₃OH 99/1). The product fractions were collected and the solvent was evaporated, yielding 5.14 g of intermediate (17) (melting point: 179.5 ℃ C.).

Intermediate (17) was prepared according to the same method but starting from 2-hydroxy-4-isopropoxy-benzoic acid methyl ester.

Intermediate (18)

Example A.10

a) Preparation ofIntermediate (19)

To a suspension of sodium hydride (0.0970 mol) in DMF (30 g) cooled with liquid nitrogen under an argon atmosphere was added methyl 2, 4-dihydroxybenzoate (0.0974 mol) in DMF (60 g) (this step was carried out in a 500 ml flask due to vigorous bubbling by hydrogen separation). The reaction mixture was slowly brought to room temperature and stirred with a magnetic stirrer. After the evolution of hydrogen had ceased, the mixture was stirred for a further 30 minutes. Bromocyclopentane (0.1220 moles) was then added and the mixture was heated at 100 ℃ for 2 hours. Evaporating DMF; will CH₂Cl₂(100 ml) was added to the residue; the precipitate is filtered off and taken up with CH₂Cl₂(2 × 10 ml) wash. To merge CH₂Cl₂The solution was evaporated in vacuo. The crude product was dissolved in methanol (20 g) and the resulting solution was crystallized at-10 deg.c (in a refrigerator) for 15 hours. The crystals were filtered off, washed with cold methanol (2 × 20 ml) and dried in vacuo to give 10.5 gIntermediate (19) (melting point: 42 ℃ -43 ℃ C.).

b) Preparation ofIntermediate (20)

Intermediate (19) (0.0797 mol) was dissolved in CH₃NH in OH₃(7N) (300 ml) and the mixture was stirred in an autoclave at 125 ℃ for 18 hours. The mixture was then cooled to room temperature, concentrated and co-evaporated with methanol. The residue was purified by column chromatography (eluent: CH)₂Cl₂/CH₃OH 97/3). The product fractions were collected and the solvent was evaporated. The pure product fractions were collected and the solvent was evaporated to give product fraction a. Passing the less pure fraction through flash chromatography (eluent: CH)₂Cl₂/CH₃OH 99/1) was further purified. The product fractions were collected and the solvent was evaporated to give product fraction B. Impure product fractions (mixed fractions) were also collected and purified by column chromatography (eluent: CH)₂Cl₂/CH₃OH 99/1) was further purified. The desired fractions were collected and the solvent was evaporated to give product fraction C. Product fractions A, B and C were combined to give 8.05 g of intermediate (20) (melting point: 146 ℃ C.).

Example A.11

Preparation ofIntermediate (21)

Intermediate (19) (0.0866 mol) was dissolved in CH₃CN (200 ml). 1-chloro-2, 5-pyrrolidinedione (0.0901 moles) was added. The reaction mixture was stirred and refluxed ± 16 hours. The solvent was evaporated. Adding CH₂Cl₂(300 ml). The mixture was washed with water (200 ml), saturated NaHCO₃The aqueous solution (200 ml) was washed and then dried (Na)₂SO₄) Filtered and the solvent evaporated. The residue is chromatographed on silica gel (eluant: heptane/ethyl acetate from 98/2 to 97-3) And (5) purifying. The desired fractions were collected and the solvent was evaporated. The residue was co-evaporated with ethyl acetate and then with CH₂Cl₂Co-evaporating. The residue was redissolved in THF (110 ml) and methanol (11 ml). To this solution was added 1M NaOH (140 ml) and the mixture was heated to reflux temperature (85 ℃) for 270 minutes. The reaction mixture was then cooled to room temperature and Et₂O (150 ml) extraction. The reaction mixture was acidified with 6N HCl (± 25 ml) and extracted with ethyl acetate (2 × 150 ml). The ethyl acetate layer was washed with brine and dried (Na)₂SO₄). The solvent of the filtrate was evaporated. The residue obtained is converted into the methyl ester. Thus, the residue was dissolved in methanol and H was added dropwise₂SO₄(0.792 ml). The reaction mixture was stirred and refluxed for 3 hours. Then add more H₂SO₄(1 ml) and the mixture was stirred and refluxed for 1 week. The solvent was evaporated. Saturated NaHCO was carefully added₃An aqueous solution. The mixture was extracted with ethyl acetate (2 × 150 ml). The separated organic layer was washed with brine and dried (Na)₂SO₄) Filtered and the solvent evaporated and reacted with CH₂Cl₂Co-evaporating. 7N NH of the residue in methanol₃(300 ml) was stirred overnight at 100 ℃ in an autoclave. The solvent is then evaporated and the residue is passed through flash column chromatography (eluent: CH)₂Cl₂/CH₃OH) purifying. The desired fractions were collected and the solvent was evaporated. The residue was crystallized from 2-propanol (90 ml). The precipitate was filtered off and the filtrate was left at room temperature overnight. The new precipitate is filtered off (residue (I)) and the solvent of the filtrate is evaporated (residue (II)). The residues (I) and (II) were combined and then recrystallized from 2-propanol. The precipitate was filtered off and recrystallized from 2-propanol, then filtered off and dried, yielding 3.69 g of intermediate (21) (melting point: 181 ℃ C.).

Example A.12

Preparation ofIntermediate (22)

Intermediate (14) (0.0144 mol) was dissolved in DMF (20 ml). 1-chloro-2, 5-pyrrolidinedione (0.0144 mol) was added in one portion. The reaction mixture was stirred at 40 ℃ for 16 hours. The solvent (DMF) was evaporated under reduced pressure. Water was added to the residue. The precipitate was filtered off and dissolved in ethyl acetate. The organic solution was dried (MgSO₄) Filtered and the solvent evaporated. The residue was purified by column chromatography on silica gel (eluent: ethyl acetate/hexane 50/50 v/v). The product fractions were collected and the solvent was evaporated, yielding 1.53 g of intermediate (22).

Example A.13

Preparation ofIntermediate (23)

Triethylamine (14 ml) was added to 1, 1-dimethylethyl 4-hydroxy-piperidine-1-carboxylate (0.0670 mol) in CH₂Cl₂(300 ml) in solution. 4-methylbenzene-sulfonyl chloride (0.0740 moles) was added slowly under nitrogen. The reaction mixture was stirred at room temperature for 2 hours. Water was added and the mixture was extracted, dried, filtered and the solvent was evaporated. The residue was dissolved in pyridine. 4-Methylbenzenesulfonyl chloride in pyridine is added at 0 ℃. The reaction mixture was stirred at room temperature under nitrogen overnight. The precipitate was filtered off and the solvent of the filtrate was evaporated. The residue (filtrate) was dissolved in CH₂Cl₂Neutralized and extracted with water. The organic layer was dried, filtered and the solvent was evaporated. After co-evaporation with toluene, the residue was dissolved in heptane and the precipitate was filtered off to give intermediate (23).

Example A.14

a) Preparation ofIntermediate (24)

The reaction was carried out under a nitrogen atmosphere. Mixing sodium chloride (0.52)5 moles) was added to cyclopentanecarboxylic acid (0.105 moles) in CH₂Cl₂(200 ml) while cooling on an ice bath. The mixture was refluxed for 2 hours and the solvent was evaporated. Dissolving the residue in CH₂Cl₂(150 mL) and 4-amino-5-chloro-2-methoxy-benzoic acid methyl ester (0.07 mol) and Et are added₃N (19.6 ml, 0.140 mol). The reaction mixture was stirred at room temperature for 1 hour. The solvent was evaporated and the residue was dissolved in CH₂Cl₂(100 ml). The organic layer was washed with water (30 ml), saturated brine (30 ml) and then dried (MgSO)₄) Filtered and the solvent evaporated. The residue is chromatographed on silica gel (eluent: petroleum ether/CH)₂Cl₂From 2/1 up to 0/1). The product fractions were collected and the solvent was evaporated, yielding 8 g of intermediate (24).

b) Preparation ofIntermediate (25)

The reaction was carried out under a nitrogen atmosphere. Intermediate (24) (0.026 mol) was dissolved in anhydrous THF (150 ml). borane/DMS (2.6 ml, 0.026 mol) was added to this solution and the mixture was stirred at room temperature for 1 hour and then refluxed for 12 hours. The mixture was cooled to room temperature and 1N HCl (50 ml) was added. The organic layer was separated and the aqueous layer was washed with CH₂Cl₂(2 × 100 ml) extraction. The combined organic layers were washed with water (100 ml), dried and evaporated. The residue is purified by column chromatography on silica gel (eluent: from petroleum ether/CH)₂Cl₂4/1 up to 100% CH₂Cl₂) And (5) purifying. The product fractions were collected and the solvent was evaporated, yielding 2.8 g of intermediate (25).

c) Preparation ofIntermediate (26)

Intermediate (25) (0.0235 mol) was dissolved in anhydrous THF (147 ml). Reacting LiOH/H₂O(147 ml) was added to this solution and the reaction mixture was stirred at 50 ℃ for 24 hours. The mixture was cooled to room temperature and the organic solvent was evaporated. The precipitate is filtered off and CH is added₂Cl₂. The mixture was acidified with HCl (concentrated) until the precipitate was completely dissolved. The aqueous layer is replaced by CH₂Cl₂(2 × 50 ml) extraction. The combined organic layers were dried, filtered and the solvent was evaporated. The residue was washed with diisopropyl ether to give 6.00 g of intermediate (26).

d) Preparation ofIntermediate (27)

The reaction was carried out under a nitrogen atmosphere. Intermediate (26) (0.0194 mol) was dissolved in anhydrous CH₂Cl₂(100 ml). 4-methylmorpholine (0.0213 mol) was added to this solution and the mixture was cooled to 0 ℃ in an ice bath. 2-methylpropyl chloroformate (0.0388 mol) was then added slowly and the reaction mixture was stirred at 0 ℃ for 30 minutes and then at room temperature for 1 hour. NH was added at 0 ℃ for 30 minutes₃Is added to the mixture and CH is subsequently added₂Cl₂Evaporation gave 10.6 g of intermediate (27).

e) Preparation ofIntermediate (28)

Intermediate (27) (0.0194 mol) was dissolved in 1-methyl-2-pyrrolidone (100 ml). Piperazine (0.194 mol) was added to this solution and the reaction mixture was stirred at 140 ℃ for 16 hours. The 1-methyl-2-pyrrolidone was then evaporated and water was added to the residue. The precipitate was filtered off and dissolved in ethyl acetate. The solution was dried, the drying agent was filtered off and the solvent was evaporated. The residue was washed with a mixture of petroleum ether and diisopropyl ether (1/1v/v) to give 3.8 g of intermediate (28).

Example A.15

a) Preparation ofIntermediate (29)

A solution of methyl 4-bromo-2-methoxybenzoate (0.1 mol) and tetrahydro-2H-pyran-4-amine (0.3 mol) in 1-methyl-2-pyrrolidone (800 ml) was stirred at 40 ℃. Adding Cs₂CO₃(0.2 mol). The reaction mixture was stirred for 5 minutes. Adding Pd₂(dba)₃(0.002 moles) and BINAP (0.003 moles). The reaction solution was degassed by alternately applying a nitrogen atmosphere and vacuum. The reaction mixture was stirred at 110 ℃ overnight. The 1-methyl-2-pyrrolidone solvent was evaporated. The residue was purified by high performance liquid chromatography. The product fractions were collected and the solvent was evaporated, yielding 13 g of intermediate (29).

b) Preparation ofIntermediate (30)

Intermediate (29) (0.049 mol) and 1-chloro-2, 5-pyrrolidinedione (0.044 mol) in CH₃The solution in CN (150 ml) was stirred and refluxed at 100 ℃ for 3 hours. The solvent was evaporated. The residue was purified by column chromatography on silica gel (gradient elution; eluent: petroleum ether/ethyl acetate from 100/0 to 50/50). The product fractions were collected and the solvent was evaporated, yielding 7 g of intermediate (30).

c) Preparation ofIntermediate (31)

Intermediate (30) (0.0233 mol) was dissolved in THF (100 ml). Addition of LiOH/H₂O (2N) (300 ml). The reaction mixture was stirred at 50 ℃ overnight. The solvent (THF) was evaporated. The pH was adjusted to 8. Adding CH₂Cl₂(250 ml). The organic layer was separated, washed with water (2 ×), dried (Na)₂SO₄) Filtration and evaporation of the solvent under reduced pressure gave 6.58 g of intermediate (31).

d) Preparation ofIntermediate (32)

Intermediate (31) (0.0227 mol) was dissolved in CH under nitrogen blanket₂Cl₂(200 ml). 4-methylmorpholine (0.02497 moles) was added. The solution was stirred for several minutes. 2-methylpropyl chloroformate (0.02497 moles) was added dropwise. The resulting reaction mixture was stirred at room temperature for ± 30 minutes. Reacting NH₃Added to the solution until all the starting material disappeared. The reaction mixture was stirred for half an hour. The solvent was evaporated. The residue was dried in vacuo to give 7 g of intermediate (32).

e) Preparation ofIntermediate (33)

Intermediate (32) (0.021 mol) was dissolved in 1-methyl-2-pyrrolidone (120 ml). Piperazine (0.21 mol) was added. The reaction mixture was stirred at 140 ℃ overnight. The solvent was evaporated. The residue was purified by column chromatography on silica gel (eluent: petroleum ether/ethyl acetate gradient from 100/0 to 50/50). The product fractions were collected and the solvent was evaporated, yielding 4.5 g of intermediate (33).

Example A.16

a) Preparation ofIntermediate (34)

HBTU (0.012 mol) was added portionwise to a mixture of 5-chloro-4- (dimethyl-amino) -2-methoxybenzoic acid (0.010 mol) in DIPEA (2 ml) and DMF (20 ml) and stirred at room temperature for 1 hour. The reaction mixture was cooled to-10 ℃ in an ice/EtOH bath. Adding NH in dioxane₃(0.5M) (40 ml) and the reaction mixture was allowed to warm to room temperature. The reaction mixture was stirred at room temperature for 30 minutes. Will dissolveThe agent evaporates. The residue was dissolved in water (100 ml) and stirred at room temperature for 30 minutes. The precipitate was filtered off. The residue is added to CH₃Stirring in CN. The precipitate was filtered off and dried (vacuum) to yield 1.7 g of intermediate (34).

b) Preparation ofIntermediate (35)

Intermediate (34) (0.0074 mol) and piperazine (0.0223 mol) in 1-methyl-2-pyrrolidone (7 ml) were stirred at 140 ℃ for 16 hours. The mixture was allowed to cool to room temperature and then poured into water (100 ml). The reaction mixture was acidified with aqueous HCl 1N. The precipitate was filtered off and dried (vacuum) to yield 1.1 g of intermediate (35).

Example A.17

a) Preparation ofIntermediate (36)

A mixture of 5-chloro-2-methoxy-4- (methylamino) benzoic acid (0.010 mol), HBTU (0.010 mol) and DIPEA (0.100 mol) in DMF (100 ml) was stirred at rt for 1 h. The reaction mixture was cooled in an ice-ethanol bath and NH was added₄HCO₃(0.050 moles). The reaction mixture was stirred at room temperature overnight. The mixture was poured into ice-water (+ -500 ml). The resulting precipitate was filtered off and dried to yield 1.760 g of intermediate (36).

b) Preparation ofIntermediate (37)

A mixture of intermediate (36) (0.010 mol) and piperazine (0.030 mol) in 1-methyl-2-pyrrolidone (10 ml) was stirred in a sealed reaction vial for 20 hours at 140 ℃. The mixture was poured into ice-water (180 ml) and then neutralized with 1N HCl. The mixture was stirred at room temperature over the weekend. The precipitate was filtered off and dried to yield 1.413 g of intermediate (37).

Example A.18

a) Preparation ofIntermediate (38)

A mixture of methyl 4-amino-2-methoxybenzoate (0.138 mol) and 1-acetyl-4-piperidone (0.14 mol) in methanol (300 ml) was hydrogenated in the presence of a thiophene solution (1 ml) with Pd/C (10%) (3 g) as a catalyst at 100 deg.C (100 bar) for 32 hours. After absorption of hydrogen (1 equivalent), the catalyst was filtered through dicalite and the solvent of the filtrate was evaporated. The residue was purified by column chromatography on silica gel (eluent: first ethyl acetate 100% and then CH)₂Cl₂/CH₃OH 95/5). The product fractions were collected and the solvent was evaporated, yielding 41.5 g of intermediate (38).

b) Preparation ofIntermediate (39)

Intermediate (38) (0.13 mol) in DMF (500 ml) was warmed to 60 ℃. 1-chloro-2, 5-pyrrolidinedione (0.13 mol) was added in portions and the reaction mixture was stirred at 60 ℃ for 3 hours. The reaction mixture was allowed to cool to room temperature and the solvent was then evaporated. The residue was stirred in water. The mixture was extracted 2 times with ethyl acetate. The solvent of the separated organic layer was evaporated. The residue was recrystallized from ethyl acetate, the precipitate was filtered off and dried (vacuum) yielding 16.5 g of intermediate (39).

c) Preparation ofIntermediate (40)

1N NaOH (aq) (0.094 mol) was added to intermediate (39) (0.047 mol) in THF (188 ml) andwater (94 ml) and then stirred at room temperature for 16 hours. The reaction mixture was acidified to pH 7. The solvent was evaporated and the residue was dissolved in water. The precipitate is filtered off and subsequently freed from CH₃And (5) recrystallizing CN. The precipitate was filtered off and dried (vacuum) to yield 7.3 g of intermediate (40).

d) Preparation ofIntermediate (41)

HBTU (0.0106 mol) was added to a mixture of intermediate (40) (0.00887 mol) in DIPEA (0.0106 mol) and DMF (20 ml) and stirred at rt for 1 h. The reaction mixture was cooled to 0-5 ℃ in an ice bath. NH in dioxane₃(0.5M) (35 mL) was added to the reaction mixture and the reaction mixture was allowed to warm to room temperature. The mixture was stirred at room temperature for 1 hour. The solvent was evaporated. The residue was dissolved in water (100 ml). The precipitate was filtered off to give fraction a. The filtrate was extracted again with ethyl acetate. The separated organic layer was combined with fraction a and the solvent was evaporated. The residue is chromatographed on silica gel (eluent: CH)₂Cl₂/(CH₃OH/NH₃) From 100/0 to 97/3). The product fractions were collected and the solvent was evaporated. Removing the residue from CH₃CN recrystallisation, the precipitate was filtered off and dried (vacuum) yielding 1.9 g of intermediate (41).

e) Preparation ofIntermediate (42)

Intermediate (41) (0.0058 mol), piperazine (0.018 mol), and 1-methyl-2-pyrrolidone (6 ml) were stirred at 140 ℃ for 16 hours. The reaction mixture was allowed to cool to room temperature and then poured into water (100 ml). The mixture was acidified with aqueous HCl 1N. The precipitate was filtered off and dried (vacuum) to yield 1.5 g of intermediate (42).

Example A.19

a) Preparation ofIntermediate (43)

The reaction was carried out under a stream of nitrogen. 1-methyl-2-pyrrolidone (40 ml) was added to a mixture of methyl 4-bromo-2-methoxybenzoate (0.040 mol) and 1- (tert-butoxycarbonyl) piperazine (0.048 mol). The mixture was stirred until completely dissolved. Then adding Cs₂CO₃(0.06 mol). The reaction mixture was heated to 40 ℃. Then Pd is added₂(dba)₃(0.0004 mol) and 98% BINAP (0.0012 mol). The reaction mixture was then stirred vigorously at 110 ℃ for 16 hours. The reaction mixture was allowed to cool to room temperature and then poured into water (200 ml). The reaction mixture was extracted with ethyl acetate (2 cycles). The solvent of the separated organic layer was evaporated. Dissolving the residue in CH₂Cl₂In (1). The mixture was then dried, filtered and CH was added₂Cl₂And (4) evaporating. The residue is chromatographed on silica gel (eluent: CH)₂Cl₂/CH₃OH from 100/0 to 98/2). The product fractions were collected and the solvent was evaporated. The residue was recrystallized from DIPE, the precipitate was filtered off and dried (vacuum) yielding 8 g of intermediate (43).

b) Preparation ofIntermediate (44)

Intermediate (43) (0.0078 mol) and 1-chloro-2, 5-pyrrolidinedione (0.0078 mol) in CH₃The mixture in CN (50 ml) was stirred at reflux for 16 hours. The reaction mixture was then cooled to room temperature and the solvent was evaporated. The residue was washed with water and CH₂Cl₂And (4) extracting. The solvent of the separated organic layer was evaporated. The residue was purified by high performance liquid chromatography. The product fractions were collected and the solvent was evaporated. The residue was dissolved in hexane. The precipitate was filtered off and dried (vacuum) to yield 1.8 g of intermediate (44).

c) Preparation ofIntermediate (45)

1N NaOH (aq) (0.00935 mol) was added to a mixture of intermediate (44) (0.00468 mol) in water (9 ml) and THF (18 ml). The reaction mixture was stirred at room temperature for 16 hours. The solvent was evaporated. The residue was stirred in water (10 ml). The mixture was acidified with 1N HCl (aq) (9 ml). The precipitate was filtered off and redissolved in DIPE. The recrystallized precipitate was then filtered off and dried (vacuum) to yield 1.5 g of intermediate (45).

d) Preparation ofIntermediate (46)

DIPEA (0.020 mol) was added to a mixture of intermediate (45) (0.0088 mol) and HBTU (0.010 mol) in DMF (50 ml) stirred at room temperature. The reaction mixture was stirred at room temperature for 1 hour. The reaction mixture was cooled in an ice-ethanol bath and NH in dioxane was added₃(0.5M) (0.020 mol). The reaction mixture was stirred for 1 hour and cooled in an ice-ethanol bath, then stirred at room temperature for 3 hours. The solvent was evaporated. The residue is washed with water/CH₃CN was stirred overnight, and the resulting precipitate was then filtered off and dried to yield 3.27 g of intermediate (46).

e) Preparation ofIntermediate (47)

A mixture of intermediate (46) (0.003 mol) and piperazine (0.009 mol) in 1-methyl-2-pyrrolidone (3 ml) was stirred in a sealed reaction vial for 16 hours at 140 ℃. The mixture was poured into water (10 ml) and then neutralized with acetic acid. The mixture was stirred at room temperature over the weekend. The precipitate was filtered off and dried to yield 0.825 g of intermediate (47).

Example A.20

Preparation ofIntermediate (48)

Reacting 4- [ [4- (aminocarbonyl) -2-chloro-5-methoxyphenyl]Amino group]A mixture of 1, 1-dimethylethyl 1-piperidine-carboxylate (0.021 mol) and piperazine (0.063 mol) in 1-methyl-2-pyrrolidone (21 ml) was stirred at 140 ℃ for 16 hours. The reaction mixture was allowed to cool to room temperature and then poured into ice-water (300 ml). The mixture was acidified with acetic acid to pH ± 4. The resulting precipitate was filtered off, washed with water and then dissolved in CH₂Cl₂In (1). The separated organic layer was dried (MgSO)₄) Filtered and the solvent evaporated. The residue was stirred in DIPE, filtered and dried in vacuo to yield 7.3 g of intermediate (48).

Example A.21

a) Preparation ofIntermediate (49)

A mixture of methyl 4-amino-2-methoxybenzoate (0.29 mol) and cyclopentanone (52 ml) in toluene (400 ml) was hydrogenated at 130 ℃ (50 kg hydrogen pressure) with Pd/C (10%) (3 g) as catalyst in the presence of 4% thiophene solution (2 ml) for 20 h. After uptake of hydrogen (1 eq), the catalyst was filtered off and the filtrate was evaporated, yielding 72 g of intermediate (49).

b) Preparation ofIntermediate (50)

A solution of intermediate (49) (0.29 mol) and NaOH (1.5 mol) in water (1000 ml) was stirred and refluxed for 2 hours. The reaction solution was allowed to cool and the resulting precipitate was filtered off and recrystallized from 2-propanol. The precipitate was filtered off and dried to yield 51.7 g of intermediate (50).

c) Preparation ofIntermediate (51)

1-chloro-2, 5-pyrrolidinedione (0.00935 mol) was added to a mixture of intermediate (50) (0.00850 mol) in DMF (40 ml) and the reaction mixture was stirred at room temperature for 16 h. The reaction mixture was poured into water (300 ml) and the reaction mixture was stirred at room temperature for 3 hours. The precipitate was filtered off, washed with water, stirred in DIPE, filtered and dried (vacuum). The resulting fraction was separated from DIPE/CH₃And (5) recrystallizing CN. The precipitate was filtered off and dried (vacuum) yielding 1.0 g of intermediate (51).

d) Preparation ofIntermediate (52)

DIPEA (0.050 mol) was added to a mixture of intermediate (51) (0.035 mol) in DMF (p.a.) (150 ml). HBTU (0.035 mol) was added portionwise to the reaction mixture and stirred at room temperature for 1 hour. The reaction mixture was cooled in an ice/EtOH bath. Adding NH in dioxane₃(0.5M) (100 ml) and stirred in an ice/EtOH bath for 1 hour. The reaction mixture was stirred at room temperature for an additional 12 hours. The solvent was evaporated. The residue was in ice water/some CH₃Stirring in CN. The precipitate was filtered off and dried to yield 11.56 g of intermediate (52).

e) Preparation ofIntermediate (53)

A mixture of intermediate (52) (0.010 mol) and piperazine (0.030 mol) in 1-methyl-2-pyrrolidone (10 ml) was stirred at 140 ℃ for 16 hours. The reaction mixture was poured into water and then neutralized with acetic acid to pH 7. The mixture was stirred at room temperature over the weekend. The precipitate was filtered off and dried to yield 1.960 g of intermediate (53).

Example A.22

a) Preparation ofIntermediate (54)

The reaction was carried out under a stream of nitrogen. 1-methyl-2-pyrrolidone (32 ml) was added to a mixture of methyl 4-bromo-2-methoxybenzoate (98%) (0.032 mol) and morpholine (0.038 mol). The mixture was stirred until completely dissolved. Then adding Cs₂CO₃(0.048 mol). The reaction mixture was heated to 40 ℃. Then Pd is added₂(dba)₃(0.00032 mol) and BINAP (98%) (0.00096 mol). The reaction mixture was then stirred at 110 ℃ for 16 hours. The reaction mixture was allowed to cool to room temperature and then poured into water (200 ml). The reaction mixture was then extracted with ethyl acetate (2 cycles). The solvent of the separated organic layer was evaporated. Dissolving the residue in CH₂Cl₂Dried, filtered and evaporated. The residue is chromatographed on silica gel (eluent: CH)₂Cl₂/CH₃OH from 100/0 to 98/2). The product fractions were collected and the solvent was evaporated. The residue was recrystallized from DIPE, the precipitate was filtered off and dried (vacuum) yielding 3.8 g of intermediate (54).

b) Preparation ofIntermediate (55)

1-chloro-2, 5-pyrrolidinedione (0.014 moles) was added to intermediate (54) (0.014 moles) in CH₃CN (100 ml). The reaction mixture was stirred at 40 ℃ for 16 hours. The solvent was evaporated. The residue was crystallized from DIPE. The precipitate was filtered off and dried to yield 2.6 g of intermediate (55).

c) Preparation ofIntermediate (56)

1N NaOH (aq) (0.0182 mol) was added to a mixture of intermediate (55) (0.0091 mol) in water (18 ml) and THF (36 ml). The reaction mixture was stirred at room temperature for 16 hours. The solvent was then evaporated. The residue was stirred in water (40 ml). The mixture was neutralized with 1N HCl (aq) (9 ml). The precipitate was filtered off and washed with water. The precipitate was then dried (vacuum) to give 2.3 g of intermediate (56).

d) Preparation ofIntermediate (57)

The reaction was carried out under a stream of nitrogen. HBTU (0.024 mol) was added to a mixture of intermediate (56) (0.020 mol) and DIPEA (0.026 mol) in DMF (100 ml) and stirred at rt for 1 h. The reaction mixture was then cooled to 5 ℃ in an ice bath. NH in dioxane₃(0.5M) (0.040 mol) was added dropwise to the reaction mixture and then stirred at room temperature for 2 hours. The solvent was evaporated and the residue was dissolved in ethyl acetate. The mixture was washed with water. The solvent of the separated organic layer was evaporated. The residue is added to CH₃CN, and the precipitate was filtered off and dried to yield 5.4 g of intermediate (57).

e) Preparation ofIntermediate (58)

A mixture of intermediate (57) (0.01 mol) and piperazine (0.03 mol) in 1-methyl-2-pyrrolidone (10 ml) was stirred at 140 ℃ for 16 hours. The reaction mixture was allowed to cool to room temperature and then poured into water (100 ml). The reaction mixture was acidified with 1N HCl (aq) to pH 6. The precipitate was filtered off and the filtration residue was stirred in DIPE. The precipitate was filtered off and dried (vacuum) to yield 1.4 g of intermediate (58).

Example A.23

a) Preparation ofIntermediate (59)

Methyl 4-amino-2-methoxybenzoate (0.3 mol) was dissolved in stirred CHCl3(1000 ml) with heating. The mixture was cooled to ± 35 ℃. Thiophosgene (0.39 mol) was added dropwise and the resulting reaction mixture was slowly heated to reflux temperature, then stirred and refluxed overnight. The reaction mixture was allowed to cool and then poured into ice-water. The organic layer was separated, washed three times with water, dried, filtered and the solvent was evaporated. The residue was purified by vacuum distillation (boiling point: 138 ℃ C. at 0.2 mm Hg) to give 56.5 g of intermediate (59).

b) Preparation ofIntermediate (60)

2, 2-diethoxyethylamine (0.24 mol) was dissolved in EtOH (250 ml). Intermediate (59) (0.24 mol) was added (exothermic temperature rose to ± 60 ℃). Precipitation was initiated. The reaction mixture was stirred overnight and allowed to cool to room temperature. The solvent was evaporated to give ± 80 g of intermediate (60).

c) Preparation ofIntermediate (61)

10% HCl (480 ml) was added to intermediate (60) (0.24 mol). The reaction mixture was warmed (up to ± 90 ℃). A solid forms. Water (300 ml) was added at. + -. 90 ℃. The reaction was allowed to cool slowly to room temperature. The resulting precipitate was filtered off, washed with water, petroleum ether and dried. A portion of this fraction (I) was reacted again in 10% HCl (480 ml). The reaction mixture was stirred and refluxed for 45 minutes, then cooled in an ice-bath and the resulting precipitate was filtered off, washed with a small amount of water, then with petroleum ether and dried to give fraction (Ia). An additional portion of fraction (I) was reacted again in 10% HCl (480 ml). The reaction mixture was stirred and refluxed for 45 minutes, then cooled in an ice-bath and the resulting precipitate was filtered off, washed with a small amount of water, then with petroleum ether and dried to give fraction (Ib). Fractions (Ia) and (Ib) were combined to give 50.7 g of intermediate (61).

d) Preparation ofIntermediate (62)

Adding HNO₃(53.13 ml) was added to water (127.8 ml). Adding NaNO₂(0.42 g) (exothermic temperature rise to. + -. 35 ℃ C.). Intermediate (61) (0.106 moles) was added portionwise at 30-35 ℃ and the temperature was kept at 30-35 ℃ (exothermic; steam, start precipitation). After the addition of intermediate (61) was complete, the reaction mixture was slowly heated to 45 ℃. The reaction mixture was stirred at 45 ℃ for 15 minutes, then allowed to cool to room temperature, and the resulting precipitate was filtered off, washed with a small amount of water, and then dissolved in a small amount of water. The mixture was basified to complete dissolution. Water was added (when needed). The mixture was acidified with HCl and the resulting precipitate was filtered off and dried to yield 18.5 g of intermediate (62).

e) Preparation ofIntermediate (63)

A mixture of intermediate (62) (0.014 mol) and DIPEA (0.031 mol) in DMF (70 ml) was stirred at rt. HBTU (0.016 mol) was added. The reaction mixture was stirred at room temperature for 1 hour, then cooled to ± 5 ℃ on an ice bath and NH in dioxane was added dropwise₃(0.5M) (56 ml). The reaction mixture was stirred at. + -. 5 ℃ for 30 minutes. The mixture was allowed to warm to room temperature and then stirred at room temperature for 2 hours. The solvent was evaporated. The residue was dissolved in water. The resulting precipitate was filtered off, washed with water and washed in CH₃CN, filtered off and dried (vacuum) to yield 2.9 g of intermediate (63).

f) Preparation ofIntermediate (64)

A mixture of intermediate (63) (0.013 mol) and piperazine (0.040 mol) in 1-methyl-2-pyrrolidone (13 ml) was stirred at 140 ℃ for 16 hours, then cooled to room temperature, poured into water (100 ml) and then acidified with 1N HCl to pH about 4. The resulting precipitate was filtered off, washed with water and dried (vacuum) to yield 1.6 g of intermediate (64).

Example A.24

a) Preparation ofIntermediate (65)

A mixture of methyl 4-amino-5-chloro-2-methoxybenzoate (0.020 mol) in 1, 3-dibromopropane (40 ml) and DIPEA (6.6 ml) was stirred at 140 ℃ for 2 hours. Reacting the mixture with CH₂Cl₂Diluted and washed with water. The separated organic layer was dried (MgSO)₄) Filtered and the solvent evaporated. The residue is chromatographed on silica gel (eluent: CH)₂Cl₂/CH₃OH from 100/0 to 99/1). The product fractions were collected and the solvent was evaporated. The residue was stirred in DIPE. The precipitate was filtered off and dried to yield 3.770 g of intermediate (65).

b) Preparation ofIntermediate (66)

Intermediate (65) (0.020 moles) in CH in methanol₃NH₂The mixture in (2M) (60 ml) was stirred in a sealed vial at 50 ℃ for 24 hours. The solvent of the reaction mixture was evaporated. Subjecting the residue to high performance liquid chromatography (with NH)₄HCO₃Buffer standard gradient elution). Collecting the productThe material was fractionated and the solvent was evaporated to give 5 g of intermediate (66).

c) Preparation ofIntermediate (67)

Will be in CH₂Cl₂Di-tert-butyl dicarbonate (0.020 mol) in (p.a.) (20 ml) was added dropwise to intermediate (66) (0.0175 mol) and DIPEA (0.020 mol) in CH₂Cl₂(80 ml) and stirred at room temperature. The reaction mixture was washed twice with ice water. The separated organic layer was dried (MgSO)₄) Filtration and evaporation of the solvent gave 7.2 g of intermediate (67).

d) Preparation ofIntermediate (68)

Intermediate (67) (0.0175 mol) and LiOH. H₂A mixture of O (0.035 mol) in water (50 ml) and dioxane (150 ml) was stirred at 50 ℃ for 2h and subsequently at room temperature overnight. The reaction mixture was stirred at 50 ℃ for another 4 hours and then at room temperature overnight. The solvent was evaporated. The residue was dissolved in ice water and then neutralized with 1N HCl (aq) (135 ml). The precipitate was filtered off and dried to yield 5.53 g of intermediate (68).

e) Preparation ofIntermediate (69)

HBTU (0.015 mol) was added portionwise to a stirred mixture of intermediate (68) (0.0147 mol) and DIPEA (0.030 mol) in DMF (100 ml) and stirred at rt for 1 h. The reaction mixture was cooled on an EtOH/ice bath. 10 ml of NH in dioxane are added in portions₃(0.5M) (0.030 mol) was added to the reaction mixture and stirred in an ice bath for 2 hours. The reaction mixture was then stirred at room temperature overnight. The solvent was evaporated. Will be provided withThe residue was combined with some CH in ice water₃CN was stirred together. The precipitate was filtered off and dried to yield 5.3 g of intermediate (69).

f) Preparation ofIntermediate (70)

A mixture of intermediate (69) (0.003 mol) and piperazine (0.009 mol) in 1-methyl-2-pyrrolidone (3 ml) was stirred at 140 ℃ for 16 hours. The reaction mixture was poured into water and then neutralized with 1N aqueous HCl and acetic acid. The mixture is treated with CH₂Cl₂And (4) extracting. The solvent of the separated aqueous layer was evaporated. The residue was purified by column chromatography on silica gel (eluent: hexane/ethyl acetate from 90/10 to 50/50). The product fractions were collected and the solvent was evaporated, yielding 0.8 g of intermediate (70).

Example A.25

a) Preparation ofIntermediate (71)

A mixture of methyl 4-amino-5-chloro-2-methoxybenzoate (0.1 mol), 2- (3-bromopropyl) -1H-isoindole-1, 3(2H) -dione (0.1 mol) and potassium iodide (catalytic amount) in DIPEA (16.5 ml) was stirred at 130 ℃ for 2 hours. The reaction mixture was stirred at room temperature overnight. 2- (3-bromopropyl) -1H-isoindole-1, 3(2H) -dione (0.056 mol) and DIPEA (10 ml) were again added to the reaction mixture and stirred at 130 ℃ for 4 hours. The reaction mixture was poured into water and then washed with water (200 ml) and CH₃CN (200 ml) was stirred. The precipitate was filtered off and dried to yield 26.1 g of intermediate (71).

b) Preparation ofIntermediate (72)

Intermediate (71) (0.027 mol) in NH₄The mixture in OH (110 ml) was stirred in a microwave at 125 ℃ for 2 hours. The solvent was evaporated. Bringing the residue to boiling CH₃OH/CH₃Stirring in CN and then overnight. The precipitate was filtered off and dried to yield 8.17 g of intermediate (72).

c) Preparation ofIntermediate (73)

Intermediate (72) (0.020 mol) was placed in acetyl acetate (40 ml) and CH₂Cl₂The mixture in (6.6 ml) was stirred at 140 ℃ for 2 hours. Reacting the mixture with CH₂Cl₂Diluted and then washed with water. The separated organic layer was dried (MgSO)₄) Filtered and the solvent evaporated. The residue is chromatographed on silica gel (eluent: CH)₂Cl₂/CH₃OH from 100/0 to 99/1). The product fractions were collected and the solvent was evaporated. The residue was stirred in DIPE. The precipitate was filtered off and dried to yield 3.76 g of intermediate (73).

d) Preparation ofIntermediate (74)

A mixture of intermediate (73) (0.013 mol) and piperazine (0.039 mol) in 1-methyl-2-pyrrolidone (13 ml) was stirred at 140 ℃ for 20 hours. More of some piperazine (0.0395 moles) was added to the reaction mixture and stirred at 140 ℃ for 24 hours. The reaction mixture was allowed to cool and then poured into ice water (200 ml). The reaction mixture was filtered and the residue was discarded. The filtrate was neutralized with acetic acid. The precipitate was filtered off and dried to yield 2.42 g of intermediate (74).

Example A.26

a) Preparation ofIntermediate (75)

BBr3(50 ml) was slowly added dropwise over 15 minutes to 2-bromo-1-chloro-4-methoxybenzene (0.448 mol) in CH stirred at-20 deg.C₂Cl₂(600 ml). The resulting reaction mixture was stirred at-20 ℃ for 20 minutes, then allowed to warm to room temperature and stirred at room temperature for 1 hour to give 85 g of intermediate (75).

b) Preparation ofIntermediate (76)

Acetyl chloride (0.036 mol) was slowly added to intermediate (75) (0.024 mol) and Et₃N (0.024 mol) in anhydrous CH₂Cl₂(60 ml). The resulting reaction mixture was stirred and refluxed for 12 hours, then cooled to room temperature, washed with 2.5M aqueous NaOH, and then washed with 2.5M aqueous HCl. The organic layer was separated and dried (Na)₂SO₄) Filtration and evaporation of the solvent gave 4.9 g of intermediate (76).

c) Preparation ofIntermediate (77)

Intermediate (76) (0.24 mol) and acetic acid-boron trifluoride complex (2: 1) (320 ml) were added to the flask. The reaction solution was stirred and refluxed for 3 hours, and then cooled to room temperature. The mixture (containing precipitate) is placed in CH₂Cl₂And water. The aqueous layer is replaced by CH₂Cl₂The extraction was performed twice. The organic layers were combined, washed with water, and then purified by preparative thin layer chromatography. The product fractions were collected and the solvent was evaporated, yielding 33 g of intermediate (77).

d) Preparation ofIntermediate (78)

Intermediate (77) (0.008 mol) was added to the flaskEr), Pd₂(dba)₃(0.366 g), BINAP (0.398 g) and Cs₂CO₃(0.012 mole). Then, tetrahydro-2H-pyran-4-amine (0.012 mol) was added. The mixture was dissolved in DMF. The reaction solution was stirred at 120 ℃ for 13 hours. The mixture was diluted with ethyl acetate (3 ×). The organic phase was washed with brine (2 ×). The combined organic layers were dried, filtered and the solvent was evaporated in vacuo. The residue was purified by preparative high performance liquid chromatography. By adding solid NaHCO₃The product fractions were basified (pH 8-9). Sodium chloride was added. The aqueous phase was washed with ethyl acetate (2 ×). The combined organic layers were washed with water (2 ×), dried (Na)₂SO₄) Filtered and the solvent evaporated. The residue was dried under high vacuum to give 2.16 g of intermediate (78).

Example A.27

Preparation ofIntermediate (79)

Intermediate (77) (0.012 mol), Pd were added to the flask₂(dba)₃(0.549 g), BINAP (0.597 g) and Cs₂CO₃(0.018 mol). 1-methylpiperazine (0.018 mol) was then added. The mixture was dissolved in DMF. The reaction solution was stirred at 120 ℃ for 13 hours. The mixture was diluted with ethyl acetate (3 ×). The organic phase was washed with brine (3 ×). The aqueous phase was washed twice with ethyl acetate. The combined organic layers were dried, filtered and the solvent was evaporated in vacuo. The residue was purified by column chromatography and then by preparative high performance liquid chromatography. The purified organic mixture was saturated with sodium chloride. The aqueous phase was washed with 1N NaOH and with ethyl acetate (2 ×). The organic layer was separated, washed with water, dried, filtered and the solvent was evaporated in vacuo. The residue was dried under high vacuum to give 1 g of intermediate (79).

Example A.28

Preparation ofIntermediate (80)

Intermediate (77) (0.012 mol), Pd were added to the flask₂(dba)₃(0.549 g), BINAP (0.597 g) and Cs₂CO₃(0.018 mol). Then, 2-methoxyethylamine (0.018 mol) was added. The mixture was dissolved in DMF. The reaction solution was stirred at 120 ℃ for 13 hours. The mixture was diluted with ethyl acetate (3 ×). The organic phase was washed with brine (3 ×). The aqueous phase was washed twice with ethyl acetate. The combined organic layers were dried, filtered and the solvent was evaporated in vacuo. The residue was purified by column chromatography and then by preparative high performance liquid chromatography. The purified organic mixture was saturated with sodium chloride. The aqueous phase was washed with 1N NaOH and with ethyl acetate (2 ×). The organic layer was separated, washed with water, dried, filtered and the solvent was evaporated in vacuo. The residue was dried under high vacuum to give 0.600 g of intermediate (80).

Example A.29

a) Preparation ofIntermediate (81)

Dissolving 2, 6-dimethyl benzoic acid (0.0666 mol) in anhydrous CH₂Cl₂(150 ml). One drop of DMF was added. Thionyl chloride (0.33 mol) was added dropwise at room temperature under nitrogen atmosphere. The reaction mixture was stirred and refluxed for 1 hour. The solvent was evaporated under reduced pressure. The residue was dried (under oil pump vacuum) to give the intermediate acid chloride, which was dissolved in anhydrous CH₂Cl₂(30 ml) to give solution (I). Adding solution (I) to 1, 4-dioxa-8-azaspiro [4.5 ]]Decane-8-ethylamine (0.0733 mol) and Et₃N (0.0666 mol) in anhydrous CH₂Cl₂(150 ml) in water. The reaction mixture was stirred and refluxed for 90 minutes. The reaction mixture was cooled and saturated NaHCO₃Aqueous solution (30 ml) was washed withWashed with water (30 ml) and dried (Na)₂SO₄) Filtration and evaporation of the solvent under reduced pressure gave 23 g of intermediate (81).

b) Preparation ofIntermediate (82)

A mixture of intermediate (81) (0.066 mol) and HCl (80 ml) in water (160 ml) was stirred and refluxed for 3 hours. The mixture was poured into ice. With Na₂CO₃Basification of the mixture to pH 9 followed by CH₂Cl₂And (4) extracting. The organic layer was filtered off and treated under reduced pressure. The residue was dried in vacuo to give 17.20 g of intermediate (82).

Example A.30

Preparation ofIntermediate (83)

1- (5-chloro-2, 4-dihydroxyphenyl) ethanone (0.0320 mol) and K₂CO₃A solution of (8.87 g) and sodium iodide (0.24 g) in DMF (60 ml) was stirred. 2-bromopropane (0.0320 mol) was added. The reaction mixture was stirred at 60 ℃ for 12 hours. The solvent was evaporated. Dissolving the residue in CH₂Cl₂Washing with water. The organic layer was evaporated. The residue was purified by column chromatography (eluent: CH)₂Cl₂Heptane 98/2). The product fractions were collected and the solvent was evaporated, yielding 0.55 g of intermediate (83).

Example A.31

Preparation ofIntermediate (84) 1- (5-chloro-2, 4-dihydroxyphenyl) ethanone (0.054 moles), intermediate (23) (0.067 moles) and NaHCO₃(9 g) dissolved in anhydrous CH₂Cl₂In (1). Will be provided withThe solvent is evaporated. The reaction mixture was heated at 120 ℃ for 8 hours, then cooled to room temperature and quenched with CH₂Cl₂And (6) diluting. The organic mixture was washed with water. The separated organic layer was dried, filtered and the solvent was evaporated, yielding 14.6 g of intermediate (84).

Example A.32

a) Preparation ofIntermediate (85)

A mixture of methyl 4-amino-5-chloro-2-methoxybenzoate (0.069 mol), 1-bromo-3-methoxypropane (0.088 mol) and DIPEA (0.069 mol) was stirred at 150 ℃ for 2 hours. The solution was then allowed to cool to room temperature and CH was added₂Cl₂. The solution was washed with water, then saturated brine and then again with water. The organic layer was dried, filtered and the solvent was evaporated. The residue is passed through column chromatography (eluent: petroleum ether/CH)₂Cl₂From 1/1 up to 0/1). The desired fractions were collected and the solvent was evaporated. The residue was dried to give 10 g of intermediate (85).

b) Preparation ofIntermediate (86)

LiOH (1M) in water (80 ml) was added to a solution of intermediate (85) (0.0279 mol) in THF (120 ml) and stirred at 40 ℃ overnight. The solvent was evaporated. The concentrate was neutralized to pH 7. The precipitate was filtered off and dried (vacuum) to yield 4.5 g of intermediate (86).

c) Preparation ofIntermediate (87)

The reaction was carried out under a stream of nitrogen. 4-methylmorpholine (0.0370 mol) was added to intermediate (86) (0.0340 mol) in anhydrous CH₂Cl₂(100 ml). 2-methylpropyl chloroformate was then slowly added to the reaction mixture and stirred at 0 ℃ for 30 minutes. The reaction mixture was stirred at room temperature for 30 minutes. NH at 0 ℃ over 30 minutes₃Added to the reaction mixture. The solvent was evaporated to yield 10 g of intermediate (87).

d) Preparation ofIntermediate (88)

A solution of intermediate (87) (0.0370 mol) and piperazine (0.0370 mol) in 1-methyl-2-pyrrolidone (200 ml) was stirred at 140 ℃ for 14 hours. The solvent was evaporated. Water was added to the crude material and the mixture was extracted 3 times with ethyl acetate. The separated organic layer was dried (MgSO)₄) Filtered and the solvent evaporated. The residue is passed through column chromatography on silica (eluent: petroleum ether/CH)₂Cl₂From 50/50 to 0/100). The desired fractions were collected and the solvent was evaporated, yielding 7 g of intermediate (88).

Example A.33

a) Preparation ofIntermediate (89)

Cyclopentanol (0.2400 mol), 4-methylbenzenesulfonyl chloride (0.1400 mol) and potassium hydroxide (0.9600 mol) were dissolved in CH₂Cl₂(200 ml). The reaction mixture was stirred at room temperature overnight. The mixture was washed with water. The organic layer was collected and the solvent was evaporated to yield 21.5 g of intermediate (89).

b) Preparation ofIntermediate (90)

1- (5-chloro-2, 4-dihydroxy-phenyl) -ethanone (0.0550 mol) was dissolved in DMSO (150 ml) and intermediate (89), (89) was added0.0660 moles) and NaHCO₃(11 g). The reaction mixture was stirred at 100 ℃ for 3 hours. Water was added. The precipitate was filtered and washed with water to give intermediate (90).

Example A.34

Preparation ofIntermediate (91)

A mixture of N- [5- (acetoxy) -2-methylphenyl ] -acetamide (0.087 mol), aluminium trichloride (0.27 mol) and sodium chloride (4 g) was stirred at 160 ℃ for 2 hours. The reaction mixture was cooled. Ice was added. The reaction mixture was extracted with dichloromethane. The organic layer was separated, dried, filtered and the solvent of the filtrate was removed by evaporation. The residue was dried in vacuo to give 9 g of intermediate (91).

Example A.35

a) Preparation ofIntermediate (92)

5-chloro-4-hydroxy-2-methoxybenzoic acid (0.0420 mol), intermediate (23) (0.0730 mol) and K₂CO₃A mixture of (10.1 g) in DMSO (200 ml) was stirred at 125 ℃. The solvent was evaporated. The residue was taken up in water and CH₂Cl₂And (6) washing. The solvent of the separated organic layer was evaporated to give 15 g of intermediate (92).

b) Preparation ofIntermediate (93)

A mixture of intermediate (92) (0.0375 mol) in 2N LiOH (500 ml) and THF (150 ml) was stirred at rt for 12 h. HCl was added to the reaction mixture until pH 6 to 7. The reaction mixture was extracted with diethyl ether. The solvent of the separated organic layer was evaporated to give 10 g of intermediate (93).

c) Preparation ofIntermediate (94)

di-1H-imidazol-1-yl-methanone (0.0143 mol) was added portionwise to intermediate (93) (0.0119 mol) in anhydrous CH₂Cl₂(150 ml) and then stirred at room temperature for 1 hour. This reaction mixture was added dropwise to NH in methanol over 30 minutes₃(7N) (200 ml). The reaction mixture was stirred at 0 ℃ for 1 hour and then at room temperature for 2 hours. The solvent was evaporated to give intermediate (94).

d) Preparation ofIntermediate (95)

A mixture of intermediate (94) (0.0130 mol) and piperazine (0.1169 mol) in 1-methyl-2-pyrrolidone (50 ml) was stirred at 140 ℃ for 16 hours. Piperazine and 1-methyl-2-pyrrolidone were evaporated (vacuum). The residue was purified by column chromatography (eluent: from petroleum ether/CH)₂Cl₂50/50 to CH₂Cl₂And then CH₂Cl₂Ethyl acetate 4/1). The desired fractions were collected and the solvent was evaporated, yielding 4.1 g of intermediate (95).

Example A.36

Preparation ofIntermediate (96)

Reacting 4- [ [4- (aminocarbonyl) -2-chloro-5-methoxyphenyl]Amino group]A mixture of 1, 1-dimethylethyl (1, 1-piperidinecarboxylate) (0.021 mol) and piperazine (0.063 mol) in 1-methyl-2-pyrrolidone (21 ml) was stirred at 140 ℃ for 16 hours. The reaction mixture was allowed to cool to room temperature and then poured into ice-water (300 ml). Acid-mixing the mixture with acetic acidThe reaction was carried out until the pH was ± 4. The resulting precipitate was filtered off, washed with water and then dissolved in CH₂Cl₂In (1). The separated organic layer was dried (MgSO)₄) Filtered and the solvent evaporated. The residue was stirred in DIPE, filtered off and dried in vacuo to yield 7.3 g of intermediate (48).

Example A.37

a) Preparation ofIntermediate (97)

A solution of 1-chloro-2, 5-pyrrolidinedione (0.2270 moles) in DMF was added dropwise to a solution of methyl 4-amino-2-hydroxybenzoate (0.2510 moles) in DMF and stirred at 40 ℃ overnight. The solvent was evaporated. The residue was purified by high performance liquid chromatography. The desired fractions were collected and taken over Na₂CO₃The aqueous solution is washed until the pH reaches 8-9. Adding CH₂Cl₂(1000 ml). The separated organic layer was washed twice with water and dried (MgSO)₄) Filtration and evaporation of the solvent gave 14 g of intermediate (97).

b) Preparation ofIntermediate (98)

Intermediate (97) (0.0496 mol) was added to CH₃With NH in OH (600 ml)₃The saturated solution was stirred in an autoclave at 125 ℃ for 14 hours. The reaction mixture was cooled to room temperature and the solvent was evaporated. The residue was washed with DIPE to give 8.5 g of intermediate (98).

Example A.38

Preparation ofIntermediate (99)

1- (2, 4-dihydroxy-5-methylphenyl) ethanone (0).0120 mol) and 2-bromopropane (0.0120 mol) were dissolved in 2-butanone (8 ml). Adding K₂CO₃(0.0210 mol), potassium iodide (catalytic amount) and DMSO (1.5 ml). The reaction mixture was stirred and refluxed for 5 hours. The mixture was cooled to 40 ℃ and diluted with water (22 ml). The product was extracted with toluene (30 ml 2 times). The toluene solution was washed with 0.5N NaOH (20 mL 2 times), 1N HCl (20 mL 1 times) and water (20 mL 2 times). The organic layer was dried, filtered and the solvent was evaporated to yield 1.3 g of intermediate (99).

Example A.39

Preparation ofIntermediate (100)

To a well stirred mixture of 20% NaOH (aq) (500 ml) and benzene (200 ml) at +5 ℃ was added 2-bromoethylamine hydrobromide (0.3300 mol). The mixture was stirred at +3 ℃ (ice-salt bath) for 10 minutes; a solution of 2, 6-dichlorobenzoyl chloride (0.3320 mol) in benzene (50 ml) was then added over 20 minutes, keeping the temperature below +7 ℃. The resulting mixture was stirred at 20 ℃ for 15 minutes. The precipitated product was filtered off, washed with water (5 × 200 ml) until pH about 7, dried and then dissolved in CH₃CN (200 ml); the solution was treated with activated carbon, concentrated to 135 g and allowed to crystallize (15 hours at +25 ℃ and then 2 hours at-10 ℃). The crystals were decanted, dried under vacuum, then triturated to a fine powder and dried at 100 ℃ and 0.05 mm Hg for 1 hour to give 82 g of intermediate (100) (melting point: 110 ℃ C.) -111 ℃.

Example A

a) Preparation ofIntermediate (101)

Intermediate (91) (0.0220 mol), 1-dimethylethyl 4-oxo-1-piperidinecarboxylate (0.0250 mol) and pyrrolidine (0)0480 mol) in methanol (150 ml) was stirred at 80 ℃ for 20 hours. The reaction mixture was cooled, filtered and the solvent was evaporated. The residue is added to CH₂Cl₂And 1N NaOH. The organic layer was separated and dried (MgSO)₄) Filtered and the solvent evaporated. The residue was suspended in DIPE and CH₃In CN, filtered off and subsequently dried in vacuo at 50 ℃ to yield 3.5 g of intermediate (101).

b) Preparation ofIntermediate (102)

Intermediate (101) (0.0130 mol) in THF (100 ml) was stirred under nitrogen. Sodium hydride (0.0170 mol) was added. The reaction mixture was stirred at 50 ℃ for 15 minutes. 1-bromo-3-methoxypropane (0.0250 mol) was added. The reaction mixture was stirred and refluxed for 20 hours. Additional 1-bromo-3-methoxypropane was added. The solvent was evaporated. The residue is added to CH₂Cl₂And water. The organic layer was separated and dried (MgSO)₄) Filtered and the solvent evaporated. The residue is chromatographed on silica gel (eluent: CH)₂Cl₂/CH₃OH 98/2). The product fractions were collected and the solvent was evaporated, yielding 4.5 g of intermediate (102).

c) Preparation ofIntermediate (103)

A mixture of intermediate (102) (0.0100 mol) in 6N HCl (17 ml) and EtOH (50 ml) was stirred and refluxed for 3 hours, then cooled and EtOH evaporated. The aqueous layer was basified with NaOH 50% (while cooling). Subjecting the mixture to CH₂Cl₂And (4) extracting. The separated organic layer was dried (MgSO)₄) Filtration and evaporation of the solvent gave 1.8 g of intermediate (103).

Example A.41

a) Preparation ofIntermediate (104)

Intermediate (101) (0.0130 moles) in THF under nitrogen. Sodium hydride (0.0120 mol) was added and stirred at 40 ℃ for 15 minutes. Methyl iodide (0.0200 mol) was then added and stirred at reflux for 20 hours. The reaction mixture was cooled and the solvent was evaporated. Dissolving the residue in CH₂Cl₂In water. The organic layer was separated and dried (MgSO)₄) Filtered and the solvent evaporated. The residue is passed through column chromatography (silica, eluent: CH)₂Cl₂/CH₃OH 98/2). The product fractions were collected and the solvent was evaporated, yielding 3 g of intermediate (104).

b) Preparation ofIntermediate (105)

A mixture of intermediate (104) (0.0075 mol) in 6N HCl (15 ml) and EtOH (50 ml) was refluxed for 20 hours. The reaction mixture was cooled, EtOH was evaporated and the residue was basified with NaOH 50% under cooling. Subjecting the mixture to CH₂Cl₂Extracting, separating the organic layer, and drying (MgSO)₄) Filtered and the solvent evaporated. The residue was suspended in DIPE, filtered off and dried in vacuo to yield 1.9 g of intermediate (105).

Example A

a) Preparation ofIntermediate (106)

A mixture of intermediate (101) (0.0050 mol) in EtOH (25 ml) was added 6N HCl (7 ml) and the resulting reaction mixture was stirred and refluxed for 1 hour. The reaction mixture was cooled and the EtOH solvent was evaporated. The aqueous acidic concentrate was basified with NaOH 50% under ice bath cooling. The solid was filtered, washed with water and dried under vacuum at 50 ℃ to give 1 g of intermediate (106).

b) Preparation ofIntermediate (107)

Di-tert-butyl dicarbonate in CHCl at 0 deg.C₃The mixture in (5 ml) was added to intermediate (106) (0.0040 mol) in CHCl₃(10 ml) of the mixture in (1). The reaction mixture was stirred for 5 minutes and washed with water. The organic layer was separated and dried (MgSO)₄) Filtered and the solvent evaporated. The residue was suspended in DIPE and dried in vacuo at 50 ℃ to give 1 g of intermediate (107).

c) Preparation ofIntermediate (108)

Cyclopentylformaldehyde (0.0080 mol) was added to intermediate (107) (0.0060 mol) in CH₂Cl₂(120 ml) and acetic acid (1.2 ml), then titanium isopropoxide (excess) was added and the reaction mixture was stirred at room temperature for 30 minutes. Adding NaBH₃CN (0.5 g) and the reaction mixture was stirred at rt for 2 h. The mixture was washed with 1N NaOH solution. The organic layer was separated and dried (MgSO)₄) Filtered and the solvent evaporated. The residue was suspended in DIPE to give 2.5 g of intermediate (108).

d) Preparation ofIntermediate (109)

Trifluoroacetic acid (20 ml) was added to intermediate (108) (0.0060 mol) in CHCl in an ice bath₃(25 ml). The reaction mixture was stirred at room temperature for 1 hour and the solvent was evaporated. The residue is taken up in CH₂Cl₂/1N NaOH, then the organic layer was separated and dried (MgSO)₄) Filtration and evaporation of the solvent gave 1.8 g of intermediate (109).

Example A.43

Preparation ofIntermediate (110)

1- (2, 4-dihydroxy-5-methylphenyl) ethanone (0.024 mol) and bromocyclopentane (2.7 ml) were dissolved in 2-butanone (16 ml). Adding K₂CO₃(5.8 g), potassium iodide (catalytic amount) and DMSO (3 ml). The reaction mixture was stirred and refluxed for 5 hours. The mixture was cooled to 40 ℃ and diluted with water (50 ml). The product was extracted with toluene (30 ml twice). The toluene solution was washed with 0.5N NaOH (20 ml twice), 1N HCl (20 ml once) and water (20 ml twice). The organic layer was separated, dried, filtered and the solvent was evaporated, yielding 3.5 g of intermediate (110).

Example A.44

Preparation ofIntermediate (111)

1- (2, 4-dihydroxy-5-methylphenyl) ethanone (0.018 mol) and intermediate (23) (0.019 mol) were dissolved in 2-butanone (12 ml). Adding K₂CO₃(4.4 g), potassium iodide (catalytic amount) and DMSO (2.2 ml). The reaction mixture was stirred and refluxed overnight. The mixture was cooled to 40 ℃ and diluted with water (50 ml). The product was extracted with toluene (30 ml twice). The toluene solution was washed with 0.5N NaOH (20 ml twice), 1N HCl (20 ml once) and water (20 ml twice). The organic layer was dried, filtered and the solvent was evaporated to give 4.9 g of intermediate (111).

Example A.45

a) Preparation ofIntermediate (112)

Intermediate (101) (0.013 mol) in THF (150 ml) was stirred under a stream of nitrogen. Sodium hydride (0.017 mole) was added. The reaction mixture was stirred at 50 ℃ for 15 minutes. 3-bromo-1-propanol (0.025 mol) was added. The reaction mixture was stirred and refluxed for 20 hours, then cooled and the solvent was evaporated. The residue is added to CH₂Cl₂And water. The organic layer was separated and dried (MgSO)₄) Filtered and the solvent of the filtrate was evaporated. The residue is chromatographed on silica gel (eluent: CH)₂Cl₂/CH₃OH 98/2). The product fractions were collected and the solvent was evaporated, yielding 1.8 g of intermediate (112).

b) Preparation ofIntermediate (113)

A mixture of intermediate (112) (0.004 mol) and 6N HCl (5.4 ml) in EtOH (20 ml) was stirred and refluxed for 2 hours. The reaction mixture was cooled and the ethanol solvent was evaporated. The aqueous concentrate was basified with NaOH 50%. The solvent was evaporated. The residue is added to CH₂Cl₂(+CH₃OH)/a little water + sodium chloride. The mixture was stirred and filtered. The separated organic layer was dried (MgSO)₄) Filtration and evaporation of the solvent gave 0.7 g of intermediate (113).

Example A.46

a) Preparation ofIntermediate (114)

After the reaction of 1- (2, 4-dihydroxy-5-methylphenyl) ethanone (12.035 mmol) and 4- [ [ (4-methylphenyl) sulfonyl group]Oxy radical]Ethyl cyclohexanecarboxylate (12.035 mmol) to a mixture of 2-butanone (8 ml) was added potassium iodide (catalytic amount), potassium carbonate (3 g) and DMSO (2 ml)). The reaction mixture was refluxed for 20 hours and cooled. Water was added to the reaction mixture, and the reaction mixture was extracted with toluene. The organic layer was washed with 1N sodium hydroxide and dried (MgSO)₄) Filtration and evaporation of the solvent gave 1.7 g of intermediate (114).

Example A.47

a) Preparation ofIntermediate (115)

2-hydroxy-5-methyl-benzoic acid (328.6 mmol) was dissolved in sulfuric acid (concentrated, 175 ml). The mixture was cooled to 0 ℃. A mixture of nitric acid (concentrated, 15.2 ml) and sulfuric acid (concentrated, 15.2 ml) was added dropwise. The mixture was stirred at 20 ℃ for a further 7 hours. The mixture was filtered off and the residue was extracted with ethyl acetate. The organic layer was washed with water (100 ml x 3) and dried (Na)₂SO₄) Filtering and evaporating. The product was dried under vacuum at 60 ℃ for 8 hours to give 30 g of intermediate (115).

b) Preparation ofIntermediate (116)

Intermediate (115) (152.17 mmol) was dissolved in CH₃OH (150 ml). Sulfuric acid (concentrated, 30 ml) was added at 0 ℃. The mixture was stirred at 85 ℃ for 16 hours. The precipitate was filtered off and dried under vacuum at 60 ℃ for 12 hours to yield 13 g of intermediate (116).

c) Preparation ofIntermediate (117)

Intermediate (116) was dissolved in THF (165 ml). Adding NaHCO₃(260.5 mmol) followed by water (165 ml). Adding Na in portions₂S₂O₄(156.3 mmol), followed by the addition of CH₃OH (165 ml). The reaction mixture is stirredFor 30 minutes. The solvent was evaporated under vacuum. The residue was dissolved in HCl (2N) solution. The residue is taken up in CH₂Cl₂(2x50 ml) extraction. The aqueous layer was washed with solid NaHCO₃Basified until pH 8 and extracted with ethyl acetate. The organic layer was washed with water (50 ml x 3) and dried (Na)₂SO₄) Filtered off and evaporated, yielding 6.6 g of intermediate (117).

d) Preparation ofIntermediate (118)

Ammonia gas was introduced into CH over 1.5 hours at-78 deg.C₃OH to obtain NH₃/CH₃OH and used immediately in this reaction. Dissolving intermediate (117) in NH₃/CH₃In OH. The mixture was stirred in an autoclave at 125 ℃ for 24 hours. The solvent was evaporated at 50 ℃ under reduced pressure. The crude material was purified by column (gradient elution: petroleum ether/ethyl acetate from 15/0 to 4/1). The desired fractions were collected and the solvent was evaporated. The product was dried under vacuum at 60 ℃ for 12 h to give 4 g of intermediate (118).

Example A.48

a) Preparation ofIntermediate (119)

Methyl 2-hydroxy-3-methoxybenzoate (137.2 mmol) was dissolved in toluene (600 ml). Sulfuryl chloride (22.2 ml) was added dropwise. The mixture was stirred at 25 ℃ for 12 hours and then heated to 60 ℃ for 6 hours. Water (100 ml) was added and Na was added₂CO₃The mixture was adjusted to pH 7. The organic layer was separated and the aqueous phase was extracted with ethyl acetate (100 ml). The organic layers were combined and washed with brine and water over Na₂SO₄Dried, filtered and evaporated. The residue was recrystallized from dioxane to give 7.6 g of pure product. The mother liquor is then evaporated to dryness and worked up on RP-18 by preparative scaleLiquid chromatography (eluent: CH)₃CN/water from 45/55 to 85/15v/v, 0.1% CF₃COOH) purification. The desired fractions were collected and the solvent was evaporated to give 9.6 g of pure product. Two batches of the sub-pure product were combined and used in the next step to give 17.2 g of intermediate (119).

b) Preparation ofIntermediate (120)

Intermediate (119) (35.09 mmol) was dissolved in CH₃Saturated ammonia in OH (400 ml). The reaction was stirred in a sealed tube at 125 ℃ for 24 hours. The solvent was evaporated. The residue was washed with a mixture of petroleum ether (20 ml) and isopropyl ether (20 ml). The precipitate was filtered off and dried to yield 6.2 g of intermediate (120).

Example A.49

a) Preparation ofIntermediate (121)

Methyl 2, 4-dihydroxybenzoate (892.9 mmol) was dissolved in DMF (1200 ml). 1-chloro-2, 5-pyrrolidinedione (892.9 mmol) was added. The reaction mixture was stirred at 50 ℃ for 4 hours. The solvent was evaporated under reduced pressure. The residue was subjected to high performance liquid chromatography (eluent: CH)₃CN/H₂O from 30/70 to 60/30, containing 0.1% CF₃COOH) purification. The product fractions were collected and the solvent was evaporated. The product was dried under vacuum at 50 ℃ for 12 h to yield 45 g of intermediate (121).

b) Preparation ofIntermediate (122)

Intermediate (121) (148.08 mmol) was dissolved in DMF (350 ml). Sodium iodide (7.40 mmol) and Na were added₂CO₃(296.15 mmoles).A solution of bromo-cyclopentane (296.15 mmol) in DMF (100 ml) was added over 30 min at 25 ℃. The mixture was stirred at 60 ℃ for 3 hours. The mixture was cooled to 25 ℃ and Na was added₂CO₃Filtering out. The filtrate was washed with 2N NaOH solution (200 ml). Subjecting the mixture to CH₂Cl₂And (4) extracting. The organic layers were combined, washed with water and dried (Na)₂SO₄) Filtered and the solvent evaporated. The residue was crystallized from ethyl acetate to yield 21.5 g of intermediate (122).

c) Preparation ofIntermediate (123)

Ammonia gas was absorbed over 30 minutes to CH in a dry ice/acetone bath₃OH to obtain NH₃/CH₃And (5) OH. Intermediate (122) (70.1823 mmol) was dissolved in NH₃/CH₃OH (1000 ml). The mixture was stirred in an autoclave at 125 ℃ and 3MPa for 24 hours. After the reaction, thin layer chromatography (petroleum ether/ethyl acetate 1/1, v/v) was performed. The solvent was evaporated under reduced pressure. The residue was washed with diisopropyl ether. The precipitate was filtered off and dried under vacuum at 50 ℃ for 24 hours to yield 16.4 g of intermediate (123).

Example A.50

a) Preparation ofIntermediate (124)

Intermediate (121) (98.7 mmol) was dissolved in DMF (200 ml). 2-bromopropane (197 mmol), sodium iodide (4.9 mmol) and K were added₂CO₃(197 mmol). The reaction mixture was stirred at 80 ℃ for 15 hours. Will K₂CO₃Filtering out. The solvent was evaporated. Dissolving the residue in CH₂Cl₂(200 ml). The mixture was washed with water (30 ml). The organics were separated and dried (Na)₂SO₄) Filtered off and evaporated. The residue was passed through a silica gel column (elution)Liquid: petroleum ether/CH₂Cl₂From 99/1 to 30/1). The product fractions were collected and dried to yield 6.1 g of intermediate (124).

b) Preparation ofIntermediate (125)

Intermediate (124) (24.9 mmol) was dissolved in CH₃Saturated ammonia in OH (180 ml). The reaction was stirred in a sealed tube at 125 ℃ for 24 hours. The solvent was evaporated. The residue was washed with a mixture of petroleum ether (20 ml) and isopropyl ether (20 ml). The precipitate was filtered off and dried to yield 5.4 g of intermediate (125).

Example A.51

a) Preparation ofIntermediate (126)

1- (2-hydroxy-5-methyl-3-nitrophenyl) ethanone (92 mmol), 1-dimethylethyl 4-oxo-1-piperidinecarboxylate (110 mmol) and pyrrolidine (220 mmol) in CH₃The mixture in OH (600 ml) was stirred at 80 ℃ for 3 hours. The reaction mixture was cooled. The solvent was evaporated. The residue was suspended in DIPE, filtered off and dried in vacuo at 50 ℃ to yield 17 g of intermediate (126).

b) Preparation ofIntermediate (127)

Intermediate (126) (44 mmol) was dissolved in methanol (300 ml) and the mixture was hydrogenated at 25 ℃ with Pd/C (10%) (3 g) as catalyst and in the presence of thiophene solution (1 ml). After uptake of hydrogen (3 equivalents), the reaction mixture was filtered to remove the catalyst. The filtrate was concentrated to dryness, suspended in DIPE, filtered and dried in vacuo at 50 ℃ to give 14 g of intermediate (127).

c) Preparation ofIntermediate (128)

Titanium isopropoxide (10 ml) was added to intermediate (127) (5.7 mmol) in CH₂Cl₂(120 ml) and CH₃COOH (1.2 ml). Cyclopentyl formaldehyde (7 mmol) was added. The reaction mixture was stirred at room temperature for 30 minutes. Adding NaBH₃CN (0.5 g). The mixture was stirred at room temperature for 20 hours, washed with water, and MgSO₄Dry, filter and evaporate the solvent. The residue was purified by column chromatography on silica gel (CH)₂Cl₂/CH₃OH 98/2). The pure fractions were collected and the solvent was evaporated, yielding 1.7 g of intermediate (128).

d) Preparation ofIntermediate (129)

CF was washed in ice bath₃COOH (13 ml) was added to intermediate (128) (4 mmol) in CHCl₃(20 ml). The reaction mixture was stirred at room temperature for 1 hour. The solvent was evaporated. The residue is taken up in CH₂Cl₂NaOH 1N extraction. The organic layer was separated and dried (MgSO)₄) Filtered and the solvent evaporated. The residue was suspended in DIPE, filtered and dried under vacuum to give 0.66 g of intermediate (129).

Example A.52

a) Preparation ofIntermediate (130)

Titanium isopropoxide (10 ml) was added to intermediate (127) (7.2 mmol) in CH₂Cl₂(120 ml) and CH₃COOH (1.2 ml). Cyclopentanone (7 mmol) was added. The reaction mixture was stirred at room temperature for 30 minA clock. Adding NaBH₃CN (0.6 g). The mixture was stirred at room temperature for 20 hours, washed with water, and MgSO₄Dry, filter and evaporate the solvent. The residue was purified by column chromatography on silica gel (CH)₂Cl₂/CH₃OH 98/2). The pure fractions were collected and the solvent was evaporated, yielding 2.8 g of intermediate (130).

b) Preparation ofIntermediate (131)

CF was washed in ice bath₃COOH (20 ml) was added to intermediate (130) (6.7 mmol) in CHCl₃(40 ml). The reaction mixture was stirred at room temperature for 1 hour and then with saturated NaHCO₃Neutralizing the solution. The organic layer was separated over MgSO₄Dry, filter and evaporate the solvent. The residue was suspended in DIPE to give 1.5 g of intermediate (131).

Example A.53

a) Preparation ofIntermediate (132)

tetrahydro-4H-pyran-4-one (10 mmol) was added to intermediate (127) (5.7 mmol) in CH₂Cl₂(120 ml) and acetic acid (1.2 ml). Titanium isopropoxide (12 mmol) was added. The reaction mixture was stirred at room temperature for 30 minutes. Adding NaBH₃CN (0.7 g). The reaction mixture was stirred at room temperature for 20 hours, washed with water and dried (MgSO)₄) Filtered and the solvent evaporated. The residue was purified by column Chromatography (CH)₂Cl₂/CH₃OH 99/1). The pure fractions were collected and the solvent was evaporated, yielding 2.2 g of intermediate (132).

b) Preparation ofIntermediate (133)

CF was washed in ice bath₃COOH (17 ml) was added to intermediate (132) (5.3 mmol) in CHCl₃(30 ml). The reaction mixture was stirred at room temperature for 1 hour. The reaction mixture was washed with saturated NaHCO₃Neutralizing the solution. The organic layer was separated over MgSO₄Dry, filter and evaporate the solvent. The residue was suspended in DIPE to give 1 g of intermediate (133).

Example A.54

a) Preparation ofIntermediate (134)

Intermediate (127) (8.7 mmol) and polyoxymethylene (0.26 g) were placed in CH₃The mixture in OH was hydrogenated with Pd/C10% (0.5 g) as catalyst and in the presence of thiophene solution (0.5 ml). After absorption of hydrogen (1 eq), the catalyst was filtered off and the filtrate was evaporated. The residue was suspended in DIPE, filtered off and dried under vacuum to yield 2 g of intermediate (134).

b) Preparation ofIntermediate (135)

Intermediate (134) (5.5 mmol) was in CHCl₃(30 ml) was stirred. CF was added on an ice bath₃COOH (18 ml). The reaction mixture was stirred on an ice bath for 1 hour. The mixture was washed with saturated NaHCO₃Neutralizing with water solution. The organic layer was separated and dried (MgSO)₄) Filtration and evaporation of the solvent gave 1.55 g of intermediate (135). This product was used without further purification.

Example A.55

a) Preparation ofIntermediate (136)

In N₂To a stirred solution of intermediate (127) (8.7 mmol) in 1, 2-dichloro-ethane (26 ml) was added 2-methoxy-1-propene (13 mmol), acetic acid (0.5 ml) and NaBH (OAc)₃(2.8 g). The reaction mixture was stirred at room temperature for 24 hours. The mixture was washed with 1N NaOH. The organic layer was separated and dried (MgSO)₄) Filtered and the solvent evaporated. The residue is chromatographed on silica gel (eluent: CH)₂Cl₂/CH₃OH 99/1). The product fractions were collected and the solvent was evaporated, yielding 1.8 g of intermediate (136).

b) Preparation ofIntermediate (137)

CHCl₃Intermediate (136) (4.7 mmol) was stirred in (25 ml). CF was added on an ice bath₃COOH (14 ml). The reaction mixture was stirred on an ice bath for 1 hour. The mixture was cooled with saturated NaHCO₃Neutralizing with water solution. The organic layer was separated and dried (MgSO)₄) Filtration and evaporation of the solvent gave 1.3 g of intermediate (137).

Example A.56

Preparation ofIntermediate (138) andintermediate (139) 4-chloro-2-methoxy-phenol (63.058 mmol) was dissolved in BF₃HOAc (72 ml). The mixture was heated to 135 ℃ for 24 hours. After cooling to 40 ℃, the mixture was poured into ice. The mixture is extracted (CH)₂Cl₂). The organic layer was washed with water. The organic layer was dried, filtered and the solvent was evaporated. The residue (10 g) was purified by column Chromatography (CH)₂Cl₂Heptane 70/30) to yield intermediate (138) and 4.9 g of intermediate (139).

Example A.57

a) Preparation ofIntermediate (140)

Intermediate (107) (8.66 mmol) was dissolved in CH₂Cl₂In (1). Triethylamine (9.526 mmol) and cyclopentane acetyl chloride (9.526 mmol) were added. After stirring at room temperature for 1 hour, water was added to the reaction mixture. The mixture is extracted (CH)₂Cl₂) And the organic layer was dried, filtered and the solvent was evaporated. The residue was used as such to give 4.5 g of intermediate (140).

b) Preparation ofIntermediate (141)

Intermediate (140) (4.38 mmol) was dissolved in EtOH (30 ml). HCl (6M, 43.804 mmol) was added. The reaction mixture was refluxed for 1 hour and cooled to room temperature. The precipitate was filtered, washed with DIPE and dried. This crude residue was used to give 1.66 g of intermediate (141).

Other intermediate compounds used in the preparation of the final compound are compounds known in the art, such as 2-hydroxy-4-methoxy-benzylamide, 2-acetyl-4-chloro-5-methylphenyl, 2-hydroxy-4-acetamido-5-chlorobenzylamide, 5-chloro-2, 4-dihydroxy-benzylamide, 2-acetyl-5-aminophenol, 2-acetylphenol, 2-acetyl-4, 5-dimethylphenol, 2-acetyl-4-methylphenol, 2-acetylbenzene-1, 3-diol, 2-acetyl-5-ethoxyphenol, 2-acetyl-4-methoxyphenol, 1- (2-ethyl-6-methoxy-phenyl) -ethanone, 1- (2-hydroxy-3, 4-dimethoxy-phenyl) -ethanone, 1- (2-hydroxy-4, 5-dimethoxyphenyl) -ethanone, 1- (4-ethoxy-2-hydroxy-3-methylphenyl) -ethanone, 1- (2-hydroxy-4, 6-dimethoxyphenyl) -ethanone, 1- (2-hydroxy-4-methoxyphenyl) -ethanone, 1- (4-fluoro-2-hydroxyphenyl) -ethanone, 4-amino-1-methylpiperidine, 1- (3, 5-dichloro-2-hydroxy-phenyl) -ethanone, 1- (5-fluoro-2-hydroxyphenyl) -ethanone, 1- (5-ethoxy-2-hydroxy-phenyl) -ethanone, 1- (3-bromo-5-chloro-2-hydroxyphenyl) -ethanone, 1- (2, 6-dihydroxy-4-methoxyphenyl) -ethanone, 1- (5-chloro-2-hydroxyphenyl) -ethanone, 1- [ 2-hydroxy-6- (2-propenyloxy) phenyl ] -ethanone, 1- (2-hydroxy-4-methylphenyl) -ethanone, 1- (4-fluoro-2-hydroxyphenyl) -ethanone, a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier thereof, 1- (3, 5-dibromo-2-hydroxyphenyl) -ethanone, 1- (3-amino-2-hydroxy-5-methylphenyl) -ethanone, 1- [ 2-hydroxy-3-methyl-4- (phenyl-methoxy) phenyl ] -ethanone, 1- (3, 5-difluoro-2-hydroxyphenyl) -ethanone, 1- (5-ethyl-2-hydroxyphenyl) -ethanone, 1- [ 2-hydroxy-5- (trifluoromethoxy) phenyl ] -ethanone, 1- (2-hydroxy-3, 6-dimethoxyphenyl) -ethanone, 1- (5-bromo-2-hydroxyphenyl) -ethanone, a pharmaceutically acceptable salt thereof, a pharmaceutically acceptable carrier thereof, and, 1- (2-hydroxy-5-nitrophenyl) -ethanone, 1- (2-fluoro-6-hydroxyphenyl) -ethanone, 1- (tert-butoxy-carbonyl) -4-aminopiperidine, N- (3-acetyl-2-hydroxy-5-methylphenyl) -carbamic acid 1, 1-dimethylethyl ester, 1- (3-chloro-2-hydroxyphenyl) -ethanone, 3-methoxy-1-propylamine, N- (4-acetyl-3-hydroxyphenyl) -acetamide, tetrahydro-2H-pyran-4-amine, trifluoroacetic anhydride, cyclopentanemethanamine hydrochloride, 2 ' -hydroxy-3 ' -methoxy-5 ' -methyl-acetophenone, 2 ', 3 ' -dihydroxy-5 ' -methyl-acetophenone, and 3-amino-5-chloro-2-hydroxy-benzylamide.

B. Preparation of the Final Compounds

Example B.1

Preparation ofCompound (1)

2, 6-Dichlorobenzoyl chloride (0.00723 mol) was added dropwise to intermediate (5) (0.00482 mol) and DIPEA (0.024 mol) in CH at room temperature over 5 minutes₂Cl₂(20 ml) and DMF (20 ml). The reaction mixture was stirred at room temperature for 3 hours. The solvent was evaporated. The residue was purified by column chromatography on silica gel (washing)Liquid removal: CH (CH)₂Cl₂/(CH₃OH/NH₃)95/5) is purified. The product fractions were collected and the solvent was evaporated. The residue was precipitated in DIPE and the precipitate was filtered off to give 0.710 g of compound (1).

Example B.2

Preparation ofCompound (2)

A mixture of compound (1) (0.0021 mol) and formaldehyde (0.124 g) in THF (50 ml) and methanol (100 ml) was hydrogenated with Pt/C (5%; 0.3 g) as a catalyst in the presence of thiophene solution (0.3 ml). After absorption of hydrogen (1 eq), the catalyst was filtered off and the filtrate was evaporated. The residue was purified by high performance liquid chromatography. The product fractions were collected and the solvent was evaporated. The residue is removed from CH₃CN recrystallizes and the precipitate is filtered off, yielding 0.143 g of compound (2).

Example B.3

Preparation ofCompound (3)

Pyrrolidine (0.0009 mol) was added to a mixture of intermediate (8) (0.009 mol) and intermediate (9) (0.009 mol) in toluene. The reaction mixture was stirred and refluxed for 16 hours using a Dean Stark water separator. Toluene was evaporated under reduced pressure. Dissolving the residue in CH₂Cl₂In (1). The mixture was washed with water, brine and then water again. The separated organic layer was dried (MgSO)₄) Filtered and the solvent evaporated under reduced pressure. The residue was purified by high performance liquid chromatography. The product fractions were collected and the solvent was evaporated. The residue was triturated under DIPE and the precipitate was filtered off and dried to yield 0.151 g of compound (3).

Example B.4

Preparation ofCompound (17)

Reacting NaBH₄(0.00055 mol) was added to a solution of compound (7) (0.00046 mol) in EtOH (5 ml) and stirred at 50 ℃ overnight. Reacting the mixture with NH₄The Cl quenched. The mixture was extracted 2 times with ethyl acetate. The separated organic layer was washed with brine and dried (MgSO)₄) Filtration and evaporation of the solvent gave 0.190 g of compound (17).

Example B.5

Preparation ofCompound (10)

Trifluoroacetic acid (0.00235 mol) and CH₂Cl₂Compound (17) (0.00039 mol) and triethylsilane (0.0039 mol) in (5 ml) were heated in a sealed tube at 60 ℃ overnight. The solvent was evaporated. Dissolving the residue in CH₂Cl₂In (1). The mixture is treated with NH₃Aqueous washing and water washing. The separated organic layer was dried (MgSO)₄) Filtered and the solvent evaporated. The residue is chromatographed on silica gel (eluent: CH)₂Cl₂/CH₃OH 97/3). The product fractions were collected and the solvent was evaporated, yielding 0.089 g of compound (10).

Example B.6

Preparation ofCompound (22)

Compound (21) (0.0167 mol), 4- [ [ (4-methylphenyl) sulfonyl group]Oxy radical]1, 1-Dimethylethyl 1, 1-piperidinecarboxylate (0.0283 moles) and K₂CO₃(6.9 g) in CH₃The mixture in CN was stirred and refluxed for 16 hours. The solvent was evaporated under reduced pressure. Water (200 ml) was added. The mixture is treated with CH₂Cl₂(3 × 150 ml) extraction. The organic layer was separated and dried (MgSO)₄) Filtered and the solvent evaporated under reduced pressure. Adding CH₃CN (50 ml). The crude oily solution was left to stand and the resulting white precipitate was filtered off, washed with DIPE and dried to yield 6.16 g of compound (22).

Example B.7

Preparation ofCompound (23)

Compound (22) (0.0092 mol) was dissolved in CH₂Cl₂(100 ml). Dropwise addition of trifluoroacetic acid CH₂Cl₂Solution (50%) (40 ml) and the reaction mixture was stirred at room temperature for 1 hour. The solvent was evaporated. Dissolving the residue in CH₂Cl₂(100 ml). The organic solution was washed with 1N NaOH and then dried (MgSO)₄) Filtered and the solvent evaporated. The residue was precipitated in DIPE, filtered off and dried (vacuum, 50 ℃) to yield 3.28 g of compound (23).

The reaction can also be carried out using HCl dissolved in isopropanol or dioxane.

Compound (99) is prepared analogously starting from compound (103) using HCl dissolved in dioxane.

Example B.8

Preparation ofCompound (26)

Compound (23) (0.00264 mol) was dissolved in methanol (20 ml). Formaldehyde (0.00792 moles) was added in one portion followed by NaBH (OAc)₃(95%) (0.0792 mol). The reaction mixture was stirred at 55 ℃ for 3 hours. Addition of additional NaBH (OAc)₃(95%) (1 g) (gas evolution over 10 minutes). The solvent was evaporated under reduced pressure. Water (100 ml) was added and 1N NaOH (up to 50 ml) was added. The mixture was extracted with ethyl acetate (3 × 200 ml). The combined organic layers were dried (MgSO)₄) Filtered and the solvent evaporated. The residue was precipitated in DIPE, filtered off and dried (50 ℃, under reduced pressure overnight) to yield 0.984 g of compound (26).

Example B.9

Preparation ofCompound (27)

Compound (23) (0.00176 mol) and DIPEA (0.00880 mol) were dissolved in CH₂Cl₂Neutralized and cooled to 0 ℃. Acetyl chloride (0.00352 mol) was added dropwise. The reaction mixture was stirred at 0 ℃ for 1 hour. The solvent was evaporated. Dissolving the residue in CH₂Cl₂Washed with water, dried, filtered and the solvent evaporated under reduced pressure. The residue (crude oil) is separated from CH₃And (4) crystallizing CN. The product fractions were collected and the solvent was evaporated. The precipitate was filtered off and dried to yield 0.825 g of compound (27).

Example B.10

Preparation ofCompound (43)

Compound (42) (0.00173 mol) was added to CH₂Cl₂The mixture in (11 ml) was stirred at room temperature. Trifluoroacetic acid (4 ml) and CH were added dropwise₂Cl₂(p.a.) (5 ml) and the reaction mixture was stirred at room temperature for 2 hours. The solvent was evaporated. Dissolving the residue in water/CH₃CN and basified with 1N NaOH. The resulting precipitate was filtered off and dried to give 0.501 g of compound (43).

Example B.11

Preparation ofCompound (45)

1- (2-hydroxyphenyl) ethanone (0.021 mol) was added to a mixture of intermediate (8) (0.013 mol) and pyrrolidine (0.043 mol) in methanol (40 ml). The reaction mixture was stirred at 80 ℃ for 12 hours and then cooled. The solvent was evaporated. Dissolving the residue in CH₂Cl₂(200 ml). The organic solution was washed with water, then brine, then dried (Na)₂SO₄) Filtered and the solvent evaporated in vacuo. The residue is chromatographed on silica gel (eluent: CH)₂Cl₂/CH₃OH 200/1). The product fractions were collected and the solvent was evaporated. Removing the residue from CH₃And (5) recrystallizing CN. The precipitate was filtered off and dried to yield 2.6 g of compound (45).

Compound (103) is similarly prepared using intermediates (8) and (84).

Example B.12

Preparation ofCompound (88)

The reaction is carried out in a microwave oven. Compound (87) (0.00549 mol), Pd (OAc)₂(0.24 g), BINAP (0.27 g) and Cs₂CO₃(0.01097 mol) was dissolved in 1-methyl-2-pyrrolidone (60 ml). Then, tetrahydro-2H-pyran-4-amine (0.01097 moles) was added. The reaction mixture was stirred at 110 ℃ for 50 minutes. The solvent was evaporated. The residue is taken up in CH₂Cl₂Diluted and then washed with water (2 ×), dried (MgSO)₄) Filtered and the solvent evaporated. The residue was purified by preparative high performance liquid chromatography. Collecting the product gradeThe solvent was evaporated. The residue was acidified with HCl/1, 4-dioxane. The salt was filtered off and dried to yield 0.48 g of compound (88).

Example B.13

Preparation ofCompound (91)

Compound (74) (0.0107 mol) was dissolved in methanol (150 ml). Raney nickel (catalytic amount) is added. The mixture was hydrogenated for 12 hours. After absorption of hydrogen (1 eq), the catalyst was filtered off and the filtrate was evaporated. The residue was purified by column chromatography on silica gel (eluent: ethyl acetate). The product fractions were collected and the solvent was evaporated. The residue was dissolved in 1, 4-dioxane and converted to the hydrochloride salt (1: 1) with HCl/2-propanol. The precipitate was filtered off and dried (vacuum) to yield 0.75 g of compound (91).

Example B.14

Preparation ofCompound (93)

The reaction was carried out under a nitrogen atmosphere. Compound (58) (0.0065 mol) was dissolved in CH₂Cl₂Neutralized and cooled to-20 ℃. Adding BBr at-20 deg.C₃(30 ml). The reaction mixture was stirred at room temperature for 12 hours. Water addition (quenching BBr)₃). By adding K₂CO₃The pH was adjusted to pH 7. Adding CH₂Cl₂. The mixture was washed with water (2 ×). The organic layer was separated and the solvent was evaporated under reduced pressure. The residue was purified by high performance liquid chromatography. The product fractions were collected and sodium chloride was added until saturated. Adding K₂CO₃The pH was brought to 9. Adding CH₂Cl₂. The organic layer was separated and the solvent was evaporated under reduced pressure. The residue was dried (vacuum oven) to give 0.350 g of compound (93).

Example B.15

Preparation ofCompound (96)

Compound (83) (0.0110 mol) was dissolved in CH₂Cl₂(400 ml). Ethyl acetate (0.0100 mol) and Et were added₃N (6.8 ml). The reaction mixture was stirred at room temperature for 1 hour. The reaction mixture was washed with water and then saturated Na₂CO₃The aqueous solution was washed and washed again with water. The organic layer was separated, dried, filtered and the solvent was evaporated to give compound (96).

Compound (100) is similarly prepared starting from compound (99).

Example B.16

Preparation ofCompound (97)

Will K₂CO₃(0.0056 mol) and 1-bromobutane (0.0022 mol) were added to a solution of compound (121) (0.0022 mol) in DMF (20 ml) and the reaction mixture was stirred at 80 ℃ for 12 hours. The reaction mixture was poured into water (30 ml). The mixture is treated with CH₂Cl₂And (4) extracting. The separated organic layer was washed with water. The solvent of the separated organic layer was evaporated. The residue was purified by high performance liquid chromatography. The product fractions were collected and the solvent was evaporated, yielding 0.6 g of compound (97).

Example B.17

Preparation ofCompound (151)

Subjecting a molecular sieve to a reaction4(15 g) and then PTSA (0.0013 mol) were added to a solution of intermediate (17) (0.0132 mol) and intermediate (82) (0.0158 mol) in toluene (30 ml) and stirred and refluxed at 50 ℃ for 12 hours. The toluene was evaporated (vacuum). Dissolving the residue in CH₂Cl₂In (1). The mixture was washed with NaOH (2N) and then brine. The separated organic layer was dried (Na)₂SO₄) Filtered and the solvent of the filtrate was evaporated. The residue is purified by column chromatography on silica (eluent: petroleum ether/EtOAc/(7N NH)₃In methanol) 100/100/1 and 0/50/1). The product fractions were collected and the solvent was evaporated. The residue was crystallized from EtOH and the precipitate was filtered off to give 2.2 g of compound (151).

Example B.18

Preparation ofCompound (106)

Compound (87) (0.0005 mol), NH₂A mixture of oh.hcl (0.0009 mol) and NaOAc (0.075 g) in EtOH was stirred and refluxed for 4 hours. After completion of the reaction, the reaction mixture was allowed to cool to room temperature. The precipitate was filtered off and crystallized from methanol to give compound (106).

Example B.19

Preparation ofCompound (108)

Compound (104) (0.0003 mol) was dissolved in THF (10 ml) and 1N HCl (0.8 ml) was added. The reaction mixture was stirred at 70 ℃ overnight. With NaHCO₃The reaction was basified, extracted and washed with water. The mixture was dried over isolute and the solvent was evaporated to give compound (108).

Example B.20

Preparation ofCompound (117)

Compound (108) (0.0015 mol) was hydrogenated in methanol (40 ml) using Pt/C (5%) (0.3 g). After uptake of hydrogen (1 eq) the reaction mixture was filtered. The solvent was evaporated. The product was purified by reverse phase high performance liquid chromatography. The residue is extracted (CH)₂Cl₂/H₂O), dried, filtered and the solvent evaporated to give compound (117).

Example B.21

Preparation ofCompound (121)

A mixture of compound (81) in methanol was hydrogenated over Raney nickel as a catalyst for 12 hours. After uptake of hydrogen (3 equivalents), the catalyst was filtered off and the solvent was evaporated, yielding 0.92 g of compound (121).

Example B.22

Preparation ofCompound (127)

A mixture of compound (120) (0.0012 moles) and iodocyclopentane (0.0050 moles) was heated in a microwave oven at 180 ℃ for 40 minutes. The crude residual reaction mixture is added to CH₂Cl₂(+ methanol) and aqueous ammonia solution. The organic layer was separated and dried (MgSO)₄) Filtered and the solvent evaporated. The reaction was carried out 4 times. The residue was purified by high performance liquid chromatography. The product fractions were collected and part of the solvent was evaporated. Adding the concentrate to CH₂Cl₂And water (containing a drop of ammonia). Will haveSeparating organic layer, drying (MgSO)₄) Filtered and the solvent evaporated. The residue was suspended in DIPE, filtered off and dried in vacuo at 50 ℃ to give 0.09 g of compound (127).

Example B.23

Preparation ofCompound (130)

To a mixture of compound (41) (0.0020 mol), sodium chloride (0.482 g) and oxone (0.0020 mol) in 2-propanone (150 ml) was added water (150 ml). The mixture was stirred at room temperature over the weekend. The solvent was evaporated. The product is treated with CH₂Cl₂Several drops of methanol and NaHCO₃And (4) extracting with an aqueous solution. The organic layer was separated, dried, filtered and the solvent was evaporated. The residue was purified by column Chromatography (CH)₂Cl₂To 90/10CH₂Cl₂/CH₃OH(NH₃) Purification). The desired fractions were collected and the solvent was evaporated to give compound (130).

Example B.24

Preparation ofCompound (141)

A mixture of intermediate (113) (0.0043 mol), intermediate (100) (0.0050 mol) and DIPEA (0.0055 mol) in DMF (25 ml) was stirred at room temperature for 24 hours. The solvent was evaporated. The residue is added to CH₂Cl₂And water. The organic layer was separated and dried (MgSO)₄) Filtered and the solvent evaporated. The residue is chromatographed on silica gel (eluent: CH)₂Cl₂/(CH₃OH/NH₃)97/3) is purified. The product fractions were collected and the solvent was evaporated. The residue was suspended in DIPE, filtered off and dried (vacuum, 40 ℃) to yield 0.4 g of compound (141).

Example B.25

Preparation ofCompound (180)

To a mixture of compound (40), sodium chloride (0.157 g) and oxone (0.0007 mol) in acetone (50 ml) was added water (50 ml). The mixture was stirred at room temperature for 3 hours. The solvent was evaporated. The product is treated with CH₂Cl₂Several drops of methanol and NaHCO₃And (4) extracting. The organic layer was separated, dried, filtered and the solvent was evaporated. The residue was purified by column chromatography (eluent: CH)₂Cl₂To 90/10CH₂Cl₂/CH₃OH(NH₃) Purification). The desired fractions were collected and the solvent was evaporated to give compound (180).

Example B.26

Preparation ofCompound (181)

A solution of compound (123) (0.0016 mol) in DMF (5 ml) was stirred under nitrogen atmosphere. Sodium hydride (0.0016 mol) was added and the reaction mixture was stirred at 50 ℃ for 10 minutes. 1-bromo-3-methoxypropane (0.0027 mol) was added and the resulting reaction mixture was stirred at 70 ℃ for 1 hour. The reaction mixture was allowed to cool. The solvent was evaporated. The residue is added to CH₂Cl₂And water. The organic layer was separated and dried (MgSO)₄) Filtration and evaporation of the solvent gave 0.8 g of compound (181).

Example B.27

Preparation ofCompound (133)

Compound (120) (0.0011 mol) and 1-bromo-3-methoxypropane (0.5 g) were stirred at 180 ℃ for 30 minutes. The mixture was allowed to cool. The mixture is added into CH₂Cl₂(+ methanol) and water. The organic layer was separated and dried (MgSO)₄) Filtered and the solvent evaporated. The residue is chromatographed on silica gel (eluent: CH)₂Cl₂/(CH₃OH/NH₃)98/2) is purified. The product fractions were collected and the solvent was evaporated. The residue was purified by reverse phase high performance liquid chromatography. The product fractions were collected and the solvent was evaporated. The residue was taken up in water (+ NH)₃drop)/CH₂Cl₂Stirring. The organic layer was separated and dried (MgSO)₄) Filtered and the solvent evaporated. The residue was suspended in DIPE and filtered off. Subjecting the fraction to high performance liquid chromatography again. The product fractions were collected and the solvent was evaporated. The residue was crystallized from ethyl acetate as the oxalate salt, then filtered and dried under vacuum at 50 ℃ to give 0.035 g of compound (133).

Example B.28

Preparation ofCompound (208)

Compound (129) (0.4 mmol) was dissolved in CH₃CN (5 ml). N-chlorosuccinimide (0.051 g) was added. The reaction mixture was stirred at 65 ℃ for 8 hours and further stirred at room temperature overnight. The solvent was evaporated and the mixture was extracted (CH)₂Cl₂/H₂O，NaHCO₃). The organic layer was dried, filtered and the solvent was evaporated. The mixture was purified by reverse phase HPLC (method a) to give 39 mg of compound (208).

Example B.29

Preparation ofCompound (189)

Bromo-cyclopentane (4.856 mmol) was added to compound (235) (3.237 mmol), K₂CO₃(4.208 mmol) and potassium iodide (catalyst) in DMF (40 ml). The reaction mixture was stirred at 60 ℃ for 22 hours. More bromo-cyclopentane (0.1 ml) was added and the reaction mixture was stirred for another 3 hours. After cooling to room temperature, water was added. The reaction mixture is extracted (CH)₂Cl₂/H₂O). The organic layer was dried, filtered and the solvent was evaporated. Subjecting the mixture to column Chromatography (CH)₂Cl₂/CH₃OH(NH₃)96/4) is purified. The desired fractions were collected and the solvent was evaporated (1 g). The residue was crystallized from DIPE to yield 530 mg of compound (189).

Example B.30

Preparation ofCompound (239)

Compound (233) (1.4 g) and polyoxymethylene (0.090 g) were placed in CH₃The mixture in OH (40 ml) was hydrogenated with Pt/C5% (0.05 g) as catalyst in the presence of thiophene solution (0.1 ml). After absorption of hydrogen (1 eq), the catalyst was filtered off and the filtrate was evaporated. The residue was purified by high performance column chromatography. The solvent was evaporated. The second fraction needs to be purified again by high performance column chromatography using method a. The solvent was evaporated. The residue was suspended in DIPE, filtered off and dried to yield 77 mg of compound (239).

Example B.31

Preparation ofCompound (222) cis andcompound (199) trans

2, 6-dichloro-N- [2- (4-oxo-1-piperidyl) ethyl]Benzylamide (9 mmol), intermediate (114) (12.485 mmol) and pyrrolidine (18 mmol) in CH₃The mixture in OH (300 ml) was stirred at 80 ℃ for 48 hours and cooled. The solvent was evaporated. The residue is added to CH₂Cl₂/H₂And (4) extracting in O. The organic layer was separated and dried (MgSO)₄) Filtered and the solvent evaporated. The residue was purified by chromatography using method a. Two fractions were collected and the solvent was evaporated. Fraction 1 was crystallized as the oxalate salt in ethyl acetate, filtered and dried under vacuum at 50 ℃ to give 305 mg of compound (222). Fraction 2 was crystallized as the oxalate salt in ethyl acetate, filtered and dried under vacuum at 50 ℃ to give 147 mg of compound (199).

Example B.32

Preparation ofCompound (223)

Reaction in N₂The reaction is carried out in an atmosphere. Compound (226) (5.58 mmol) was dissolved in acetic acid (dry glacial acetic acid, 30 ml) and cyclopentanone (39.06 mmol) was added. The mixture was stirred at 25 ℃ for 2 hours. Add NaBH portion by portion₃CN (33.5 mmol). The reaction mixture was stirred at 25 ℃ for 2 hours. An additional 500 mg of cyclopentanone was added and the reaction was stirred at 25 ℃ for an additional 1 hour. Water (30 ml) and CH were added₂Cl₂(100 ml) and passing over solid Na₂CO₃The solution was adjusted to pH 8. The solution was then filtered to remove inorganic salts. Using CH for the aqueous phase₂Cl₂And extracting again. The organic layers were combined, washed with brine and water, over Na₂SO₄Dried, filtered and evaporated. The residue is chromatographed on RP-18 by preparative high performance liquid chromatography (eluent: CH)₃CN/H₂O from 45/55 to 85/15v/v, 0.1% CF₃COOH) purification. The desired fractions were collected and purified with NaHCO₃Basified to pH 8 and extracted twice with ethyl acetate. The organic layers were combined, washed twice with water and evaporatedPassing through Na₂SO₄Dried, filtered and evaporated to give the pure product. The product was further dried under vacuum at 60 ℃ for 12 hours to give 1.16 g of compound (223).

Example B.33

Preparation ofCompound (205)

The reaction is carried out in a microwave oven. Compound (226) (6.20103 mmol), 1' -oxybis [ 2-bromo-ethane ]]A mixture of (31.0051 mmol), potassium carbonate (12.4021 mmol) and sodium iodide (0.6201 mmol) in DMF (30 ml) was stirred at 140 ℃ for 40 min. The potassium carbonate was filtered off. The filtrate was concentrated and dissolved in CH₃OH and decolorized by activated carbon. The mixture was filtered and concentrated. The residue was purified by preparative high performance liquid Chromatography (CH)₃CN/H₂O from 15/85 to 40/60, containing 0.1% CF₃COOH) purification. The product was taken up with solid Na₂CO₃Basified until pH 9 is reached and CH is added₃CN was evaporated. The resulting precipitate was filtered off and washed with water (2 times 10 ml), filtered and then separated from CH₃And (5) recrystallizing CN. The precipitate was filtered off and dried under vacuum at 80 ℃ for 1 hour to give 563.36 mg of compound (205).

Tables F-1, F-2, F-3, F-4 and F-5 list compounds prepared according to one of the above examples.

TABLE F-1

TABLE F-2:

table F-3:

table F-4:

C₂H₂O₄represents an ethanedioic acid salt

Table F-5:

C. analysis section

C.1 melting Point

The melting points (m.p.) of various compounds were determined. Values are peak or melting ranges and are obtained with experimental uncertainties typically associated with this analytical method. The melting points reported in Table F-6 were obtained using DSC823e (Mettler-Toledo; temperature gradient of 30 ℃/min, max. 400 ℃), Diamond DSC (PerkinElmer; temperature gradient of 10 ℃/min, max. 300 ℃), WRS-2A melting point apparatus (Shanghai Precision and Scientific Instrument Co. Ltd.; linear heat rise 0.2-5.0 ℃/min, max. 300 ℃) or Kofler hot bench (consisting of a hot plate with linear temperature gradient, a sliding pointer and a temperature scale of a thermometer).

Table F-6: melting Point (m.p. is defined as melting point)

(*): measurement Using WRS-2A melting Point apparatus

(**): using Diamond DSC from PerkinElmer and measuring with a temperature gradient of 10 ℃ per minute

C.2 LCMS method

General procedure A

The HPLC measurements were performed using an Alliance HT 2790(Waters) system including a quadrupole pump with degasser, an auto injector, a column oven (set at 40 ℃ unless otherwise specified), a Diode Array Detector (DAD) and columns as indicated in the methods below. The flow from the column was split to the MS spectrometer. The MS detector was equipped with an electrospray ionization source. Mass spectra were acquired by scanning from 100 to 1000 in 1 second using a dwell time of 0.1 second. The capillary needle voltage was 3kV and the source temperature was maintained at 140 ℃. Nitrogen was used as the sparging gas. Data acquisition was performed using a Waters-Micromass MassLynx-Openlynx data system.

LCMS general method B

The HPLC measurements were performed using an Agilent 1100 module, including a pump, a Diode Array Detector (DAD) (wavelength 220 nm used), a column heater, and columns as indicated in the methods below. Flow from the column was split to Agilent MSD Series G1946C and G1956A. The MS detector was equipped with API-ES (atmospheric pressure electrospray ionization). Mass spectra were obtained by scanning from 100 to 1000. The capillary needle voltage was 2500V for positive ionization mode and 3000V for negative ionization mode. The fragmentation voltage (fragmentation voltage) was 50V. The temperature of the drying gas was maintained at 350 ℃ under a flow of 10 l/min.

LCMS general method C

The LC measurements were performed using the Acquisty UPLC (Waters) system, including a binary pump, a sample organizer, a column heater (set at 55 ℃), a Diode Array Detector (DAD), and columns as indicated in the methods below. The flow from the column was split to the MS spectrometer. The MS detector was equipped with an electrospray ionization source. Mass spectra were obtained using a dwell time of 0.02 seconds to scan from 100 to 1000 in 0.18 seconds. The capillary needle voltage was 3.5kV and the source temperature was maintained at 140 ℃. Nitrogen was used as the sparging gas. Data acquisition was performed using a Waters-MicromassMassLynx-Openlynx data system.

LCMS-method 1

In addition to general procedure a: reverse phase HPLC was performed on an Xterra MS C18 column (3.5 μm, 4.6 × 100 mm) with a flow rate of 1.6 ml/min. Gradient conditions were run from 100% A to 1% A, 49% B and 50% C for 6.5 minutes, to 1% A and 99% B for 1 minute using three mobile phases (mobile phase A: 95% 25mM ammonium acetate + 5% acetonitrile; mobile phase B: acetonitrile; mobile phase C: methanol) and these conditions were maintained for 1 minute and equilibrated with 100% A for an additional 1.5 minutes. An injection volume of 10 microliters was used. The cone voltage was 10V for positive ionization mode and 20V for negative ionization mode.

LCMS-method 2

In addition to general procedure a: the column heater was set at 60 ℃. Reverse phase HPLC was performed on an Xterra MS C18 column (3.5 μm, 4.6 × 100 mm) with a flow rate of 1.6 ml/min. Gradient conditions were run from 100% A to 50% B and 50% C for 6.5 minutes using three mobile phases (mobile phase A: 95% 25mM ammonium acetate + 5% acetonitrile; mobile phase B: acetonitrile; mobile phase C: methanol) to 100% B at 0.5 minutes and holding these conditions for 1 minute and equilibrating with 100% A for an additional 1.5 minutes. An injection volume of 10 microliters was used. The cone voltage was 10V for positive ionization mode and 20V for negative ionization mode.

LCMS-method 3

In addition to general procedure a: reverse phase HPLC was performed on an Atlantis C18 column (3.5 μm, 4.6 × 100 mm) with a flow rate of 1.6 ml/min. Two mobile phases were used (mobile phase A: 70% methanol + 30% H)₂O; mobile phase B: at H₂0.1% formic acid in O/methanol 95/5) gradient conditions were run from 100% B to 5% B + 95% a12 min. An injection volume of 10 microliters was used. The cone voltage was 10V for positive ionization mode and 20V for negative ionization mode.

LCMS-method 4

In addition to general procedure B: reverse phase HPLC was performed on YMC-Pack ODS-AQ, 50X2.0 mm 5 μm column with a flow rate of 0.8 ml/min. Two mobile phases (mobile phase A: water with 0.1% TFA; mobile phase B: acetonitrile with 0.05% TFA) were used. First, 100% A was held for 1 minute. Then a gradient was applied to 40% a and 60% B for 4 minutes and held for 2.5 minutes. Typically an injection volume of 2 microliters is used. The furnace temperature was 50 ℃. (MS polarity: Positive).

LCMS-method 5

In addition to general procedure B: reverse phase HPLC was performed on YMC-Pack ODS-AQ, 50X2.0 mm 5 μm column with a flow rate of 0.8 ml/min. Two mobile phases (mobile phase A: water with 0.1% TFA; mobile phase B: acetonitrile with 0.05% TFA) were used. First, 90% A and 10% B were held for 0.8 min. Then a gradient was applied to 20% a and 80% B for 3.7 minutes and held for 3 minutes. Typically an injection volume of 2 microliters is used. The furnace temperature was 50 ℃. (MS polarity: Positive).

LCMS-method 6

In addition to general procedure C: reversed phase uplc (ultra Performance liquid chromatography) was performed on a bridged ethylsiloxane/silica hybrid (hybrid) (BEH) C18 column (1.7 μm, 2.1x50 mm; Waters Acquity) with a flow rate of 0.8 ml/min. Two mobile phases are used (mobile phase A: in H)₂0.1% formic acid in O/methanol 95/5; mobile phase B: methanol) to operate gradient conditions from 95% a and 5% B to 5% a and 95% B for 1.3 minutes and hold for 0.2 minutes. An injection volume of 0.5 microliters was used. The cone voltage was 10V for positive ionization mode and 20V for negative ionization mode.

LCMS-method 9

In addition to general procedure B: reverse phase HPLC was performed on an Ultimate XB-C18, 50X2.1 mm 5 micron column with a flow rate of 0.8 ml/min. Two mobile phases were used (mobile phase C: 10 mmol/l NH)₄HCO₃(ii) a Mobile phase D: acetonitrile). First, 90% C and 10% D were held for 0.8 min. Then a gradient was applied to 20% C and 80% D for 3.7 minutes and held for 3 minutes. Typically an injection volume of 2 microliters is used. The furnace temperature was 50 ℃. (MS polarity: Positive).

LCMS-method 10

In addition to general procedure a: the column heater was set at 45 ℃. Reverse phase HPLC was performed on an Xterra MS C18 column (3.5 μm, 4.6 × 100 mm) with a flow rate of 1.6 ml/min. Three mobile phases were used (mobile phase A: in H)₂0.1% formic acid in O/methanol 95/5; mobile phase B: acetonitrile; mobile phase C: methanol) to operate gradient conditions from 100% a to 1% a, 49% B and 50% C for 7 minutes and maintain these conditions for 1 minute. An injection volume of 10 microliters was used. The cone voltage was 10V for positive ionization mode.

TABLE F-7: LC/MS data-residence time (R in minutes for the principal component)_t)，(MH)⁺Peak (free base), LCMS method

Co.Nr.	Rt	(MH)+	LCMS method
				2	0.82	527	6
3	0.99	525	6
				4	0.88	491	6
5	0.98	517	6
				6	0.85	477	6
9	0.74	463	6
				13	0.95	492	6
14	1.05	518	6
				15	1.14	552	6
16	1.04	526	6
				23	0.63	567	6
26	0.60	581	6
				36	6.43	551	1
41	4.10	448	1
				42	6.13	654	1
52	4.29	493	4
				54	3.46	566	5
55	3.54	666	1
				56	0.99	491	6
57	0.83	493	6
				58	0.86	463	6
59	0.97	502	6
				60	2.77	579	5
61	4.24	579	9
				62	3.69	501	5
66	5.53	540	1
				73	3.89	462	4
82	2.93	502	5
				88	6.34	566	5
90	3.92	554	4
				94	4.37	490	1
99	4.26	566	1
				100	5.36	608	1

Co.Nr.	Rt	(MH)+	LCMS method
				103	6.64	666	1
104	4.58	610	2
				105	4.60	610	2
107	6.22	525	1
				108	5.93	592	1
162	4.60	569	4
				164	4.50	500	4
109	5.45	527	2
				112	6.35	509	2
114	5.91	552	2
				166	4.29	501	4
115	4.59	609	1
				116	6.98	534	1
117	5.64	594	2
				168	3.69	526	4
169	4.77	568	4
				170	4.60	567	4
119	7.67	551	3
				122	6.96	535	1
180	5.32	498	3
				124	6.56	511	1
125	6.22	553	1
				127	4.37	530	10
129	5.81	516	1
				131	5.84	591	1
136	5.78	593	1
				140	6.95	537	1
141	5.40	520	3
				143	6.24	533	1
146	5.18	588	1
				147	5.53	476	3
149	6.37	544	1

D. Pharmacological data

D.1 hENT1-NBMPR-ERYTH assay

The term from Moravek Biochemicals (Brea, CA) was used in the binding assay³H]NBMPR (19.9 Ci/mmol) determines the affinity of a compound for the human ENT1 transporter. Erythrocytes were isolated from freshly isolated EDTA anticoagulated human blood. 4 ml of human whole blood was diluted in 11 ml of wash buffer (20mM MOPS, 130mM NaCl, pH 7.4) and centrifuged at 800g for 5 minutes. The red blood cells were washed twice with wash buffer by centrifugation at 800g for 5 minutes and then resuspended in wash buffer to the original whole blood volume and stored at-80 ℃. Used in assay buffer (20mM Tris, 140mM NaCl, 5mM NaCl, 2mM MgCl)₂0.1mM EDTA, 5mM glucose, pH 7.4) washed red blood cells diluted 1: 200 and 1nM radioligand were subjected to binding experiments at apparent binding equilibrium (incubation for 30 min at room temperature). Will testThe compounds were pre-incubated with red blood cells for 30 minutes at room temperature. Nonspecific binding was estimated in the presence of 1 μ M trifluoxazine. Incubations were stopped by rapid filtration using a Unifilter-96 GF/C filter plate in a 96-well Perkinelmer Filtermate harvester followed by three washes with ice-cold assay buffer. Bound radioactivity was determined by liquid scintillation counter (Topcount, (PerkinElmer)). pIC₅₀＝-log(IC₅₀) The values are given in Table F-8 below.

TABLE F-8: pIC for the inhibition of the ENT1 transporter₅₀

Co.No.	pIC50
		1	7.05
2	8.19
		3	8.37
4	7.91
		5	8.39
6	8.35
		7	7.50
8	7.35
		9	7.28
10	7.73
		11	8.31
12	8.72
		13	8.50
14	8.80
		15	9.10
16	8.90
		18	8.12

Co.No.	pIC50
		19	8.56
20	7.31
		21	6.08
22	9.65
		23	7.99
24	9.10
		25	8.48
26	7.96
		27	9.11
28	8.48
		29	8.19
30	8.73
		31	9.52
32	7.80
		33	9.70
34	7.84
		35	7.08

Co.No.	pIC50
		36	9.00
37	8.69
		38	6.91
39	7.96
		40	7.22
42	9.30
		43	7.05
44	8.11
		45	6.38
46	7.69
		47	7.25
48	6.47
		49	7.82
50	6.60
		51	6.11
52	7.75
		53	6.19

Co.No.	pIC50
		54	8.56
55	8.60
		56	8.79
57	6.40
		58	7.05
59	9.11
		60	7.20
61	7.33
		62	8.36
63	6.81
		64	7.15
65	8.32
		66	8.74
67	7.82
		68	7.38
69	7.26
		70	7.37

Co.No.	pIC50
		71	7.27
72	8.30
		73	7.24
74	8.94
		76	7.23
77	7.50
		78	7.44
79	7.07
		80	8.19
81	6.36
		82	7.88
83	8.17
		85	6.84
86	8.73
		89	8.34
90	9.22
		91	8.54
92	9.17
		93	6.92
94	7.38
		95	8.78
96	9.11
		97	7.92
98	6.83
		99	8.38
100	9.23
		101	9.53
102	8.26
		104	9.09
105	7.87
		108	10.35
109	8.81
		110	7.24
111	7.74
		112	9.73
114	8.55
		115	8.39
116	9.18
		117	9.23
120	6.90
		122	9.29
124	9.12

Co.No.	pIC50
		125	8.87
126	9.33
		127	9.11
130	8.01
		131	9.49
133	8.74
		135	7.59
136	8.92
		137	9.41
138	9.31
		140	10.10
141	8.43
		143	8.54
144	8.74
		145	8.82
146	9.22
		147	8.26
148	8.10
		149	9.46
151	7.64
		152	7.93
153	7.26
		154	7.92
155	7.71
		156	8.69
157	7.96
		158	7.91
161	8.43
		162	8.07
163	9.30
		164	7.91
165	6.49
		186	7.38
167	7.55
		168	6.57
169	8.35
		170	7.65
172	6.91
		177	8.42
178	7.18
		179	8.01
185	8.995

Co.No.	pIC50
		186	7.615
187	8.815
		188	8.07
189	8.59
		190	8.605
193	7.54
		194	7.945
195	8.105
		196	7.73
197	8.335
		198	7.725
199	9.465
		200	8.175
201	8.74
		202	7.39
203	7.52
		204	7.45
205	6.97
		206	8.375
207	8.275
		208	8.82
209	8.91
		210	8.535
211	8.055
		212	7.345
213	8.44
		214	7.91
215	7.115
		217	7.66
218	8.025
		219	8.49
220	8.405
		221	7.375
222	9.275
		223	8.42
224	8.485
		225	8.71
226	7.58
		227
228	7.72
		229	7.16
230	7.18

Co.No.	pIC50
		231	8.77
232	8.06
		233	8
234	7.735
		235	7.005
236	9.335
		238	7.925
239	8.815
		240	8.16
241	7.4
		242	8.28
243	8.215
		244	7.09
245	7.51
		246	8.44
247	7.56
		248	6.6
249	8.31
		250	7.95
251	7.675
		252	7
253	7.29
		254	7.895
255	5.76
		256	6.5
257	7.36
		258	7.33
259	6.29
		260	6.98
261	6.68
		262	6.88
263	7.05
		264	7.905
265	6.22
		266	7.32
267	5.94
		268	6.19
269	7.21
		270	5.42

Claims (12)

1. A compound of the formula (I),

including any stereochemically isomeric form thereof, wherein

-A-B-represents

Wherein n is an integer 0;

R¹³represents a hydroxyl group;

R¹⁴represents hydrogen;

in the divalent groups (a-4), (a-5) and (a-6), any hydrogen atom on the same or different carbon atom may be replaced by a halogen;

R¹and R²Each independently selected from halogen or C_1-6An alkyl group;

R³、R⁴、R⁵and R⁶Each independently selected from hydrogen, halogen, C_1-6Alkyl radical, NO₂Ring of¹Or X-R⁸Wherein X represents O or NR⁹，

Wherein R is⁹Is hydrogen, C_1-6Alkyl or C_1-6Alkoxy radical C_1-6Alkyl radical, and

wherein R is⁸Is hydrogen, C_1-6Alkyl radical, C_2-6Alkenyl radical, C_3-6Cycloalkyl, polyhaloC_1-6Alkyl radical, C_1-6Alkylcarbonyl, polyhaloC_1-6Alkyl carbonyl, ring²、-(C＝O)-(CH₂)_m-Ring²Or

Via hydroxy, C_3-6Cycloalkyl radical, C_1-6Alkoxy-or phenyl-substituted C_1-6An alkyl group;

m is an integer of 0 or 1;

ring (C)¹Is selected from

Wherein R is¹⁰Is hydrogen, C_1-6Alkyl or C_1-6An alkoxycarbonyl group; and is

Ring (C)²Is selected from

Wherein R is¹⁰Is hydrogen, C_1-6Alkyl radical, C_1-6Alkylcarbonyl or C_1-6An alkoxycarbonyl group;

or a pharmaceutically acceptable acid addition salt thereof.

2. The compound of claim 1, wherein R¹And R²Are all halogens.

3. The compound of claim 1, wherein R¹And R²Are all C_1-4An alkyl group.

4. A compound according to claim 1, wherein the-a-B-group represents (a-1).

5. A compound according to claim 1, wherein the-a-B-group represents (a-2).

6. The compound of claim 1, wherein R³Is hydrogen and R⁵Is X-R⁸。

7. The compound of claim 1, wherein the compound is N- (2- {7- [ (1-acetylpiperidin-4-yl) oxy ] -6-chloro-4-oxo-3, 4-dihydro-1 ' H-spiro [ chromene-2, 4 ' -piperidin ] -1 ' -yl } ethyl) -2, 6-dichlorobenzylamide, or a pharmaceutically acceptable acid addition salt thereof.

8. The compound of claim 1, wherein the compound is N- [2- (7-amino-8-chloro-4-oxo-3, 4-dihydro-1' H-spiro [1, 3-benzoOxazine-2, 4' -piperidines]-1' -yl) ethyl]-2, 6-dichlorobenzylamide, or a pharmaceutically acceptable acid addition salt thereof.

9. A pharmaceutical composition comprising a pharmaceutically acceptable carrier and a therapeutically active amount of a compound according to any one of claims 1 to 8.

10. A process for preparing a pharmaceutical composition according to claim 9, wherein a therapeutically active amount of a compound according to any one of claims 1 to 8 is intimately mixed with a pharmaceutically acceptable carrier.

11. Use of a compound according to any one of claims 1 to 8 in the manufacture of a medicament for treating a disease associated with inhibition of ENT1 receptors in an animal.

12. A process for the preparation of a compound of formula (I) as claimed in claim 1, wherein

a) N-alkylating a compound of formula (III) with a compound of formula (II) in a reaction-inert solvent and optionally in the presence of a suitable base

Or; b) reacting a compound of formula (IV) with a compound of formula (V) in a reaction-inert solvent, and optionally in the presence of a suitable base

Or c) converting the compounds of formula (I) with each other according to conversion reactions known in the art; or, if desired, converting a compound of formula (I) into a pharmaceutically acceptable acid addition salt or, conversely, converting an acid addition salt of a compound of formula (I) with a base into the free base form; and, if desired, preparing stereochemically isomeric forms thereof, wherein-A-B-, R¹、R²、R³、R⁴、R⁵And R⁶As claimed in claim1, W is a leaving group.