RU2418786C2 - Enantiomerically pure flavone derivatives for treatment of proliferative disorders and methods of their obtaining - Google Patents

Abstract

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention relates to enantiomerically pure compound, represented by the following formula (I) or to its pharmaceutically acceptable salt

where R₁ represents phenyl, which is substituted with 2 different substituents selected from: halogen, fluoromethyl, difluoromethyl and trifluoromethyl, on condition that if one of substituents on phenyl group represents halogen, the other substituent is not halogen; R₂ and R₃ each is independently selected from: hydroxyl and OR₈; where R₈ represents substituted or unsubstituted C₁-C₁₀-alkyl, C₁-C₄-alkanoyl, substituted or unsubstituted aroyl; R₄ represents C₁-C₄-alkylene hydroxyl; and R₉ represents hydrogen or C₁-C₄-alkyl, to its enantiomerically pure intermediates, to methods of obtaining enantiomerically pure compound (I) and its intermediates, and to pharmaceutical composition, which includes enantiomerically pure compound of formula (I), as well as to application of formula (I) compound for manufacturing medication for cancer treatment.

EFFECT: obtained and described is novel enantiomerically pure compound, which can be usefull in treatment of diseases or disorders, mediated by inhibiting of cyclin-dependent kinase, for instance, cancer.

9 cl, 98 ex, 2 tbl, 3 dwg

Description

Field of Invention

This invention relates to enantiomerically pure (+) - trans-enantiomer of pyrrolidines substituted with flavones represented by formula (I), its enantiomerically pure intermediates, methods for their corresponding preparation, their use as active ingredients in pharmaceutical preparations, in particular for the treatment of proliferative disorders, for example cancer, and pharmaceutical compositions including them.

BACKGROUND OF THE INVENTION

Cell cycle regulators have gained widespread importance in proliferative diseases. Cyclin-dependent kinases (CDKs) are a family of enzymes that become activated in specific phases of the cell cycle.

A wide variety of diseases is characterized by uncontrolled cell proliferation, which results from some damage in regulatory pathways in the cell cycle [for example, overexpression of cyclins or deletion of genes encoding CKI (CDK inhibitory proteins)]. Overexpression of cyclin D1 leads to deregulation of CDK4-D1 kinase activity and, thus, contributes to uncontrolled cell proliferation. With knowledge of the role of CDK in cell cycle regulation and the discovery that approximately 90% of all neoplasias are associated with hyperactivation of CDK leading to inactivation of the Rb pathway (retinoblastoma), CDKs are beneficial targets for the development of anticancer drugs.

The prominent role of CDK / cyclin kinase complexes, in particular CDK4 / cyclin D1 kinase complexes, in inducing cell proliferation and their deregulation in tumors makes them ideal targets for the development of highly specific antiproliferative agents.

Flavopiridol and its analogues are well known as effective CDK inhibitors and provide a possible approach to antiproliferative therapy.

U.S. Patent Application Published Pending, No. 2004/0106581, describes novel compounds used in the inhibition of CDK and having good selectivity and cytotoxicity compared to various proliferative cell lines. This patent application is included in this description with reference in its entirety. The new compounds, which are disclosed in the aforementioned patent application, have two chiral centers and, therefore, can exist as four enantiomers, for example, (+) - trans, (-) - trans, (+) - cis and (-) - cis .

It is well known in the art that the enantiomers of a given chemical compound, despite the division of identical chemical compositions, can have different effects when placed in biological systems. It often happens that one enantiomer provides beneficial effects, while the other enantiomer can be harmful or inert. Thus, the benefits that are associated with the introduction of the racemic mixture can be maintained by using a single enantiomer of the compound without associated harmful side effects.

In particular, the studies found that the inhibitory activity of CDK and antiproliferative activity are due to the (+) - trans-enantiomer of pyrrolidines substituted with flavones represented by formula (I), and not (-) - trans-enantiomer.

It will be desirable for the (+) - trans enantiomer of pyrrolidine substituted flavones, compounds of formula (I), to be available in a form substantially free of its (-) - trans enantiomer to provide a safe and effective way to treat diseases and disorders mediated inhibiting cyclin-dependent kinases (CDKs); and also treating diseases associated with excessive cell proliferation, such as cancer.

In view of these findings, these inventors have developed a new method for producing enantiomerically pure (+) - trans-enantiomer of pyrrolidines substituted with flavones represented by formula (I).

SUMMARY OF THE INVENTION

This invention relates to an enantiomerically pure (+) - trans-enantiomer of pyrrolidines substituted with flavones represented by the following formula (I) (hereinafter referred to as a compound of formula (I)) and a metabolite, prodrug, pharmaceutically acceptable salt, pharmaceutically acceptable solvate or polymorph thereof:

,

,

where the definitions of R ¹ , R ² , R ³ , R ⁴ and R ⁹ are given further in the detailed description.

This invention also relates to a method for producing an enantiomerically pure compound of formula (I), or a metabolite, prodrug, pharmaceutically acceptable salt or a pharmaceutically acceptable solvate thereof. The method produces (+) - trans-enantiomer with high chemical and enantiomeric purity and in high yield.

This invention also relates to intermediates useful in a method for preparing an enantiomerically pure compound of formula (I).

This invention additionally relates to a method for re-dissolving a compound of formula (VIA) and its derivatives, this compound are key intermediates for obtaining an enantiomerically pure compound of formula (I).

The present invention further relates to a pharmaceutical composition for treating a disease or disorder in a mammal mediated by inhibition of CDK, comprising a therapeutically effective amount of an enantiomerically pure compound of formula (I) or a salt, solvate or prodrug thereof as an active ingredient and a pharmaceutically acceptable carrier.

The invention further relates to a pharmaceutical composition for treating a disease or disorder in a mammal associated with excessive cell proliferation, comprising a therapeutically effective amount of an enantiomerically pure compound of formula (I) or a salt, solvate or prodrug thereof as an active ingredient and a pharmaceutically acceptable carrier.

The present invention further relates to a method for treating a disease or disorder mediated by inhibition of CDK in a mammal, if necessary, comprising administering to said mammal an effective amount of an enantiomerically pure (+) trans-enantiomer of a compound of formula (I) or a metabolite, prodrug, pharmaceutically acceptable salt, pharmaceutically acceptable MES or its polymorph.

This invention further relates to a method for treating a disease or disorder in a mammal associated with excessive cell proliferation, comprising administering a therapeutically effective amount of an enantiomerically pure compound of formula (I) or a salt, solvate or prodrug thereof as an active ingredient and a pharmaceutically acceptable carrier.

BRIEF DESCRIPTION OF THE FIGURES

Figure 1: Chiral HPLC (high performance liquid chromatography) (Chiralcel OD-H column (250 × 4.6 mm)) gave (-) - trans- [1-methyl-3- (2,4,6-trimethoxyphenyl) -pyrrolidine -2-yl] methanol as described in example 5.

Figure 2: Chiral HPLC (Chiralcel OD-H column (250 × 4.6 mm)) obtained (-) - trans- [1-methyl-3- (2,4,6-trimethoxyphenyl) pyrrolidin-2-yl] -methanol as described in example 98 (prior art method).

Figure 3: Chiral HPLC (Chiralcel OD-H column (250 × 4.6 mm)) obtained (+) - trans-2- (2-chlorophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1- methylpyrrolidin-3-yl) -chromen-4-one hydrochloride as described in Example 10.

DETAILED DESCRIPTION OF THE INVENTION

The compounds of formula (I) according to this invention are used in the inhibition of CDK, in particular CDK4 / cyclin D1 complexes and are used in antiproliferative therapies for diseases characterized by excessive cell growth, for example cancer, cardiovascular disorders, inflammation and arthritis, nephrological disorders, parasitology, psoriasis, Alzheimer's disease, immunological disorders, including unwanted leukocyte proliferation, restenosis and other proliferative smooth muscle disorders, viral infections and fungal infections ii.

The present invention is particularly directed to an enantiomerically pure compound of formula (I) and to a metabolite, prodrug, pharmaceutically acceptable salt, pharmaceutically acceptable solvate or polymorph thereof. As mentioned above, the compounds that are disclosed in U.S. Patent Application Published Pending Application No. 2004/0106581 have two chiral centers and, therefore, can exist as four enantiomers, namely (+) trans, (- ) -trans, (+) - cis and (-) - cis. More particularly, this invention relates to an enantiomerically pure (+) - trans-enantiomer of pyrrolidines substituted with flavones represented by formula (I).

The effectiveness of the racemic compound disclosed in published US patent application No. 2004/0106581 and its individual enantiomers have been studied in detail by these inventors. It was observed that only the (+) - trans enantiomer is active. This invention represents the position of the enantiomerically pure (+) - trans enantiomer represented by formula (I), possibly thereby making it possible to reduce the required dose of the drug and its side effects.

Described below are the definitions of various expressions used to describe the compounds of this invention. These definitions apply to expressions because they are applied throughout the description (unless they are otherwise limited in meaning) either individually or as part of a larger group. They should not be interpreted literally. They are not basic definitions and are relevant only for this application.

The expression "flavon" or "chromon" or their analogues mean compounds that can be represented by the following basic structure:

As used herein, the term “alkyl” refers to a radical of saturated aliphatic groups, including straight chain and branched chain alkyl groups. Moreover, unless otherwise specified, the term “alkyl” includes unsubstituted alkyl groups as well as alkyl groups which are substituted with one or more different substituents.

Examples of alkyl residues containing from 1 to 20 carbon atoms are: methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl, dodecyl, tetradecyl, hexadecyl, octadecyl and eicosyl, n-isomers of all of these residues, isopropyl, isobutyl, 1-methylbutyl, isopentyl, neopentyl, 2,2-dimethylbutyl, 2-methylpentyl, 3-methylpentyl, isohexyl, 2,3,4-trimethylhexyl, isodecyl, sec-butyl or tert-butyl.

The expression “alkoxy,” as used herein, refers to an alkyl group, as defined above, having an oxygen radical attached to it. Representative alkoxyl groups include methoxy, ethoxy, propoxy, t-butoxy and the like.

The expression "alkenyl" refers to unsaturated analogs of aliphatic groups in length and possible substitution of the alkyls described above, but which contain at least one double, for example, 1, 2 or 3 double bonds, provided that the double bonds are not located in within the cyclic alkyl group in such a way that leads to an aromatic system. Examples of alkenyl groups include vinyl, 1-propenyl, 2-propenyl, 2-butenyl, 2-methyl-1-propenyl or 3-methyl-2-butenyl.

Moreover, unless otherwise indicated, the expressions "alkenyl" include unsubstituted and substituted alkenyl groups.

The expression “aryl,” as used herein, refers to monocyclic or polycyclic hydrocarbon groups having up to 14 ring carbon atoms in which there is at least one carbocyclic ring that has a conjugated pi-electronic system. Examples of (C ₆ -C ₁₄ ) -aryl residues are phenyl, naphthyl, biphenyl, fluorenyl or anthracenyl. Examples of (C ₆ -C ₁₀ ) -aryl residues are phenyl or naphthyl. Unless otherwise specified, and regardless of any specific substituents associated with aryl groups that are indicated in the definition of compounds of formula (I), aryl residues, for example phenyl, naphthyl or fluorenyl, can mainly be unsubstituted or substituted with one or more for example 1, 2, 3, 4 or 5, identical or different substituents. Unless otherwise specified, substituents that may be present on substituted aryl groups are: halogen, alkyl, alkenyl, alkynyl, fluoromethyl, difluoromethyl, trifluoromethyl, hydroxyl, aryloxy, amino, cyano, nitro, thiol, imine, amide or carbonyl (e.g. , carboxyl, formate, urea, ester, ketone or aldehyde), sulfhydryl, silyl ether, thiocarbonyl (e.g., thia complex, thioacetate or thioformate), sulfonyl, amino acid ester or heterocycle which is saturated, partially unsaturated or aromatic. Aryl residues may be linked in the required position, and in substituted aryl residues the substituent may be placed in the required position.

The term “heterocycle” refers to a saturated, partially unsaturated or aromatic ring containing five or six ring atoms, of which 1, 2 or 3 are identical or different heteroatoms selected from: nitrogen, oxygen and sulfur, and where the heterocycle is substituted or unsubstituted 1, 2, or 3 identical or different substituents selected from: halogen, nitro, cyano, SR ⁷ , C ₁ -C ₄ alkyl, fluoromethyl, difluoromethyl, trifluoromethyl, C ₁ -C ₄ alkoxy, hydroxyl, carboxy, C ₁ -C ₄ alkoxycarbonyl, C ₁ -C ₄ alkylene hydroxyl, CONH ₂ , CONR ₅ R ₆ , SO ₂ NR ₅ R ₆ , cycloalkyl, NR ₅ R ₆ .

The expression “heteroatom”, as used herein, means an atom of any element, with the exception of carbon or hydrogen. Preferred heteroatoms are nitrogen, oxygen, sulfur and phosphorus.

Halogen is fluoro, chloro, bromo or iodo, preferably fluoro, chloro or bromo.

It will be understood that “substitution” or “substituted” includes the implicit condition that such substitution is in accordance with the allowed valency of the substituted atom and substituent, and also represents a stable compound that does not undergo rapid transformation, for example, rearrangement, cyclization, cleavage, and etc.

It should be noted that any heteroatom with an unsaturated valency is believed to have a hydrogen atom to saturate the valency.

The expression "racemic" or "racemate" and other similar expressions refer mainly to equimolar proportions of the (+) - enantiomer and (-) - enantiomer of the compound in the composition.

The expression "enantiomerically pure" refers to a compound or compounds that or which are in enantiomeric excess of more than 95%. Preferably, the enantiomeric excess is greater than 97%. More preferably, the enantiomeric excess is greater than 99%.

The expression "enantiomeric excess" refers to the difference between the amount of one enantiomer and the amount of another enantiomer that are present in the product mixture. Thus, for example, an enantiomeric excess of 96% refers to a product mixture having 98% of one enantiomer and 2% of another enantiomer.

The expression “substantially free” means that the amount of (+) - trans enantiomer predominates in the composition compared to the (-) - trans enantiomer of the compound of formula (I). More specifically, this means that the amount of (+) - trans enantiomer compared to (-) - trans enantiomer by weight is at least about 95%, more preferably more than 97%.

As used herein, the term “prodrugs” refers to forms of a compound that are transformed in vivo to the parent compound of this invention, for example, by hydrolysis in blood. Thus, prodrugs are compounds that carry groups that are removed by biotransformation before illustration from the pharmacological action. Such groups include portions that are rapidly cleaved in vivo from a compound carrying them; this compound, after cleavage, remains or becomes pharmacologically active. Such metabolically cleavable groups form a class well known to a medical technician.

The expression "pharmaceutically acceptable salt (s)", as used herein, unless otherwise indicated, includes salts of the basic or acidic groups of the compounds of the invention, these groups are capable of forming salts. In the case where the compounds of formula (I) contain one or more acidic or basic groups, the invention also includes their corresponding pharmaceutically or toxicologically acceptable salts. Compounds of formula (I) that contain one or more basic groups, i.e. groups that can attach a proton, and can be used according to this invention in the form of their additional salts with non-toxic inorganic or organic acids. Examples of suitable inorganic acids include: boric acid, perchloric acid, hydrochloric acid, hydrobromic acid, sulfuric acid, sulfamic acid, phosphoric acid, nitric acid and other inorganic acids known to the person skilled in the art. Examples of suitable organic acids include: acetic acid, propionic acid, succinic acid, glycolic acid, gluconic acid, stearic acid, lactic acid, malic acid, tartaric acid, citric acid, ascorbic acid, pamic acid, maleic acid, hydroxy maleic acid, hydroxy maleic acid , glutamic acid, benzoic acid, salicylic acid, sulfanilic acid, 2-acetoxybenzoic acid, fumaric acid, toluenesulfonic acid, methanesulfonic acid, ethanedisulfonic acid, oxalic acid, isethionic acid, ketoglutaric acid, benzenesulfonic acid, glycerophosphoric acid and other organic acids known to the person skilled in the art. Compounds of formula (I) which contain acid groups can be used according to the invention, for example, as alkali metal salts, such as lithium (Li), sodium (Na) and potassium (K) salts. The pharmaceutically acceptable salts of this invention can be synthesized from a test compound that contains a basic or acidic moiety by conventional chemical methods. Typically, salts are prepared by contacting the free base or acid with stoichiometric amounts or with an excess of the necessary salt-forming inorganic or organic acid or base in a suitable solvent or dispersant, or anion exchange, or cation exchange with other salts. Suitable solvents are, for example, ethyl acetate, ether, alcohols, acetone, THF (tetrahydrofuran), dioxane or mixtures of these solvents.

The invention furthermore includes all solvates of compounds of formula (I), for example hydrates or adducts with alcohols, as well as derivatives and prodrugs of compounds of formula (I) that contain physiologically acceptable and cleavable groups, for example esters and amides.

Various polymorphs of compounds of formula (I) forming part of the invention can be obtained by crystallization of compounds of formula (I) under various conditions. For example, various commonly used solvents or mixtures thereof are used for crystallization; crystallization at various temperatures; various types of cooling, ranging from very fast to very slow cooling during crystallization. Polymorphs can also be obtained by heating or melting the compound, followed by gradual or rapid cooling. The presence of polymorphs can be determined by IR spectroscopy, solid state NMR, differential scanning calorimetry, powder X-ray diffraction, or other similar techniques.

Where stereochemistry is depicted in structures, it is represented relatively, and not in absolute configuration.

This invention relates to an enantiomerically pure compound of formula (I), to a metabolite, prodrug, pharmaceutically acceptable salt, pharmaceutically acceptable solvate or polymorph thereof.

,

,

Where

R ₁ represents phenyl which is substituted or unsubstituted with 1, 2, or 3 identical or different substituents selected from: halogen, nitro, cyano, C ₁ -C ₄ alkyl, fluoromethyl, difluoromethyl, trifluoromethyl, C ₁ -C ₄ - alkoxy, hydroxyl, carboxy, C ₁ -C ₄ alkoxycarbonyl, C ₁ -C ₄ alkylene hydroxyl, CONH ₂ , CONR ₅ R ₆ , SO ₂ NR ₅ R ₆ , cycloalkyl, NR ₅ R ₆ , SR ₇ ; or is a heterocycle that is a saturated, partially unsaturated or aromatic ring containing five or six ring atoms, of which 1, 2 or 3 are identical or different heteroatoms selected from: nitrogen, oxygen and sulfur, and where the heterocycle is substituted or unsubstituted 1, 2 or 3 identical or different substituents selected from: halogen, nitro, cyano, C ₁ -C ₄ alkyl, fluoromethyl, difluoromethyl, trifluoromethyl, C ₁ -C ₄ alkoxy, hydroxyl, carboxy, C ₁ -C _4- alkoxycarbonyl, C ₁ -C ₄ -alkylene oxyl, CONH ₂ , CONR ₅ R ₆ , SO ₂ NR ₅ R ₆ , cycloalkyl, NR ₅ R ₆ and SR ₇ ;

where R ₅ and R _{6 are} each independently selected from: hydrogen, C ₁ -C ₄ alkyl, C ₁ -C ₄ alkoxycarbonyl, C ₁ -C ₄ alkylacarbonyl, aryl; or R ₅ and R ₆ together with the nitrogen atom to which they are bonded may form a five or six membered ring, which may optionally contain an additional heteroatom; and

R ₇ may be selected from hydrogen, C ₁ -C ₄ alkyl, aryl, SR ₁₀ , where R ₁₀ may be selected from C ₁ -C ₄ alkyl or aryl;

R ₂ and R _{3 are} each independently selected from: halogen, hydroxyl and OR ₈ ; where R ₈ represents a substituted or unsubstituted C ₁ -C ₁₀ alkyl, C ₁ -C ₄ alkanoyl, substituted or unsubstituted aroyl;

R ₄ is C ₁ -C _4. Alkylene hydroxyl; and

R ₉ represents hydrogen or C ₁ -C ₄ alkyl.

In alternative preferred embodiments of the enantiomerically pure compounds of formula (I), the groups R ₁ -R ₃ and R ₉ , independently of one another, have the preferred meanings given below:

R ¹ represents phenyl which is substituted or unsubstituted with 1, 2 or 3 identical or different substituents selected from: halogen, nitro, cyano; C ₁ -C ₄ alkyl, fluoromethyl, difluoromethyl, trifluoromethyl, C ₁ -C ₄ alkoxy, hydroxyl, carboxy, C ₁ -C ₄ alkoxycarbonyl, C ₁ -C ₄ alkylene hydroxyl, CONH ₂ , CONR ₅ R ₆ , SO ₂ NR ₅ R ₆ , cycloalkyl, NR ₅ R ₆ and SR ₇ ;

R ₂ and R ₃ are OR ₈ , where R _{8 is} as defined and is the same or different from R ₂ and R ₃ ;

R ⁹ represents C ₁ -C ₄ alkyl;

or a metabolite, prodrug, pharmaceutically acceptable salt, pharmaceutically acceptable solvate or polymorph of preferred embodiments.

According to this invention, the most preferred enantiomerically pure compound of formula (I), or a salt or prodrug thereof, is selected from:

(+) - trans-2- (2-Chlorophenyl) -8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -5,7-dimethoxychromen-4-one;

(+) - trans-2- (2-chlorophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one;

(+) - trans-2- (2-chlorophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one hydrochloride;

(+) - trans-2- (2-Chloro-5-fluorophenyl) -8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -5,7-dimethoxychromen-4-one;

(+) - trans-2- (2-Chloro-5-fluorophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one;

(+) - trans-2- (2-Chloro-5-fluorophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one hydrochloride;

(+) - trans-2- (2-Bromo-5-fluorophenyl) -8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -5,7-dimethoxychromen-4-one;

(+) - trans-2- (2-Bromo-5-fluorophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one;

(+) - trans-2- (2-bromo-5-fluorophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one hydrochloride;

(+) - trans-2- (2-bromo-5-fluorophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one methanesulfonate;

(+) - trans-2- (2,4-dichloro-5-fluorophenyl) -8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -5,7-dimethoxychromen-4-one;

(+) - trans-2- (2,4-dichloro-5-fluorophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one;

(+) - trans-2- (2,4-Dichloro-5-fluorophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one hydrochloride;

(+) - trans-2- (2,4-dichloro-5-fluorophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one methanesulfonate;

(+) - trans-2- (2,4-Dichloro-5-fluorophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one citrate;

(+) - trans-2- (2,4-dichloro-5-fluorophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one tartrate;

(+) - trans-2- (2,4-dichloro-5-fluorophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one maleate;

(+) - trans-2- (2,4-dichloro-5-fluorophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one acetate;

(+) - trans-2- (2,4-Dichloro-5-fluorophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one sulfate;

(+) - trans-2- (2,4-dichloro-5-fluorophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one nitrate;

(+) - trans-2- (2,4-dichlorophenyl) -8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -5,7-dimethoxychromen-4-one;

(+) - trans-2- (2,4-dichlorophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one;

(+) - trans-2- (2,4-dichlorophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one hydrochloride;

(+) - trans-2- (4-bromo-2-chlorophenyl) -8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -5,7-dimethoxychromen-4-one;

(+) - trans-2- (4-bromo-2-chlorophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one;

(+) - trans-2- (4-bromo-2-chlorophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrlidin-3-yl) -chromen-4-one hydrochloride;

(+) - trans-2- (2-Chloro-4-cyanophenyl) -8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -5,7-dimethoxychromen-4-one;

(+) - trans-2- (2-Chloro-4-cyanophenyl) -8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -5,7-dimethoxychromen-4-one hydrochloride;

(+) - trans-2- (2-Chloro-4-cyanophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one;

(+) - trans-2- (2-Chloro-4-cyanophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one hydrochloride;

(+) - trans-2- (2-Chloro-4-trifluoromethylphenyl) -8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -5,7-dimethoxychromen-4-one;

(+) - trans-2- (2-Chloro-4-trifluoromethylphenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one;

(+) - trans-2- (2-Chloro-4-trifluoromethylphenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one hydrochloride;

(+) - trans-2- (2-Chloro-4-nitrophenyl) -8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -5,7-dimethoxychromen-4-one;

(+) - trans-2- (2-Chloro-4-nitrophenyl) -8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -5,7-dimethoxychromen-4-one hydrochloride;

(+) - trans-2- (2-Chloro-4-nitrophenyl) -8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -5,7-dimethoxychromen-4-one gluconate;

(+) - trans-2- (2-Chloro-4-nitrophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one;

(+) - trans-2- (2-Chloro-4-nitrophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one hydrochloride;

(+) - trans-2- (4-amino-2-chlorophenyl) -8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -5,7-dimethoxychromen-4-one;

(+) - trans-2- (4-amino-2-chlorophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one;

(+) - trans-2- (2-Chloro-4-methylaminophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one;

(+) - trans-2- (4-amino-2-chlorophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one hydrochloride;

(+) - trans-2- (2-Chloro-4-methylaminophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one hydrochloride;

(+) - trans-2- (2-Chloro-4-pyrrolidin-1-yl-phenyl) -8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -5,7-dimethoxychromen-4-one;

(+) - trans-2- (2-Chloro-4-pyrrolidin-1-yl-phenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chrome-4- she;

(+) - trans-2- (2-Chloro-4-pyrrolidin-1-yl-phenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chrome-4- it is hydrochloride;

(+) - trans-2- (2-Chloro-4-isopropylaminophenyl) -8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -5,7-dimethoxychromen-4-one;

(+) - trans-2- (2-Chloro-4-isopropylaminophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one;

(+) - trans-2- (2-Chloro-4-isopropylaminophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one hydrochloride;

(+) - trans-2- (2,4-dibromophenyl) -8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -5,7-dimethoxychromen-4-one;

(+) - trans-2- (2,4-dibromophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one;

(+) - trans-2- (2,4-dibromophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one hydrochloride;

(+) - trans-2- (2-Bromo-4-chlorophenyl) -8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -5,7-dimethoxychromen-4-one;

(+) - trans-2- (2-Bromo-4-chlorophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one;

(+) - trans-2- (2-bromo-4-chlorophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one hydrochloride;

(+) - trans-2- (2-Chlorophenyl) -5-hydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -7-methoxychromen-4-one;

(+) - trans-2- (2-chlorophenyl) -5-hydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -7-methoxychromen-4-one hydrochloride;

(+) - trans-2- (2-chlorophenyl) -5-hydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -7-methoxychromen-4-one gluconate;

(+) - trans-2- (2-Chlorophenyl) -5-hydroxy-8- (2-hydroxymethyl) -1-methylpyrrolidin-3-yl) -7- (2-methoxyethoxymethoxy) -chromen-4-one;

(+) - trans-2- (2-Chlorophenyl) -5-hydroxy-8- (2-hydroxymethyl) -1-methylpyrrolidin-3-yl) -7- (2-methoxyethoxymethoxy) -chromen-4-one gluconate;

(+) - trans-2- (2-chlorophenyl) -8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -5,7-dimethoxychromen-4-one hydrochloride;

(+) - trans-2- (2-chlorophenyl) -8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -5,7-dimethoxychromen-4-one citrate;

(+) - trans-2- (2-chlorophenyl) -8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -5,7-dimethoxychromen-4-one tartrate;

(+) - trans-2- (2-chlorophenyl) -8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -5,7-dimethoxychromen-4-one glutamate;

(+) - trans-2- (2-chlorophenyl) -8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -5,7-dimethoxychromen-4-one maleate;

(+) - trans-2- (2-chlorophenyl) -8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -5,7-dimethoxychromen-4-one nitrate;

(+) - trans-2- (2-chlorophenyl) -8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -5,7-dimethoxychromen-4-one acetate;

(+) - trans-2- (2-chlorophenyl) -8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -5,7-dimethoxychromen-4-one gluconate;

(+) - trans-acetic acid 3- [2- (2-chlorophenyl) -5,7-dihydroxy-4-oxo-4H-chromen-8-yl] -1-methylpyrrolidin-2-yl methyl ester;

(+) - trans-acetic acid 3- [2- (2-chlorophenyl) -5,7-dihydroxy-4-oxo-4H-chromen-8-yl] -1-methylpyrrolidin-2-yl methyl hydrochloride ester;

(+) - trans-Acetic acid of 8- (2-acetoxymethylpyrrolidin-3-yl) -2- (2-chlorophenyl) -5-hydroxy-4-oxo-4H-chromene-7-yl ester;

(+) - trans-Benzoic acid 8- (2-benzoyloxymethylpyrrolidin-3-yl) -2- (2-chloro phenyl) -5-hydroxy-4-oxo-4H-chromene-7-yl ester;

(+) - trans-Benzoic acid 8- (2-benzoyloxymethylpyrrolidin-3-yl) -2- (2-chloro-phenyl) -5-hydroxy-4-oxo-4H-chromen-7-yl hydrochloride ester;

(+) - trans-Octanoic acid of 2- (2-chlorophenyl) -5-hydroxy-8- (1-methyl-2-octanoyloxymethylpyrrolidin-3-yl) -4-oxo-4H-chromene-7-yl ester;

(+) - trans-Acetic acid 3- [2- (2-Chlorophenyl) -5-hydroxy-7-methoxy-4-oxo-4H-chromen-8-yl] -1-methylpyrrolidin-2-yl-methyl complex ether; and

(+) - trans-acetic acid 3- [2- (2-chlorophenyl) -5-hydroxy-7- (2-methoxy ethoxymethoxy) -4-oxo-4H-chromen-8-yl] -1-methylpyrrolidin-2 -yl-methyl ester.

In another embodiment of the present invention, a method for producing an enantiomerically pure compound of formula (I) is provided. One approach known in the art provides one enantiomer by resolving the racemic mixture of the compound at the end of a series of reactions. This approach is intended to separate enantiomers several steps earlier in the row and, thus, to provide an effective method. This inventor has established a racemic compound represented by the formula (VIA) as ideal for resolution.

,

,

where R ₂ , R ₃ , R ₄ and R ₉ are as defined, or a pharmaceutically acceptable salt thereof.

The re-dissolution method of the present invention involves the reaction of a racemic compound represented by the formula ((±) -VIA) with a chiral additive in the presence of a simple solvent to obtain the corresponding mixture of diastereomeric salts of (+) - and (-) - enantiomers of the compound of formula (VIA), separating the corresponding diastereomeric salts and treating the diastereomeric salt of the (-) - enantiomer with a base to obtain the free base of the necessary (-) - enantiomer of the compound represented by the following formula.

In the re-dissolution method of this invention, the chiral additive is selected from: (-) - dibenzoyl tartaric acid ((-) - DBTA), (+) - dibutyl tartaric acid, (-) - dibutyl tartaric acid, (+) - ketopinic acids, (-) - ketopinic acid, (+) - camphor-10-sulfonic acid, (-) - camphor-10-sulfonic acid, (+) camphoric acid and (-) - camphoric acid. The most preferred chiral additive is (-) - dibenzoyl tartaric acid ((-) - DBTA).

The simple solvent used in the re-dissolution step can be selected from: methanol, isopropanol, diisopropyl ether, ethyl acetate and chloroform. The most preferred solvent is methanol.

The base may be selected from: sodium bicarbonate, sodium carbonate and potassium carbonate. The most preferred base is sodium carbonate.

More particularly, a method was provided for re-dissolving a racemic compound of formula (VIA), where R ₂ and R ₃ are methoxy, R ₉ is methyl and R ₄ is hydroxymethyl, the compound is chemically known as (±) -trans- [1-methyl -3- (2,4,6-trimethoxyphenyl) pyrrolidin-2-yl] methanol (hereinafter defined as compound A).

A method for re-dissolving Compound A is described in U.S. Patent Application Laid-Open No. 2004/010658. This method includes many stages of crystallization and recrystallization. The dissolved compound (-) - trans- [1-methyl-3- (2,4,6-trimethoxyphenyl) -pyrrolidin-2-yl] -methanol (defined here as compound B) was obtained in 22-30% yield and reported optical rotation of -17.7 ° (c = 1.1, methanol).

In a preferred embodiment of the invention, a method comprising re-dissolving an intermediate of a compound of formula (VIA), especially compound A, involves reacting compound A with (-) - DBTA ((-) - dibenzoyl tartaric acid) as a chiral auxiliary methanol to obtain crystallized salts (+) - and (-) - trans- [1-methyl-3- (2,4,6-trimethoxyphenyl) pyrrolidin-2-yl] methanol dibenzoyltartrate. A preferred salt of the (-) - dibenzoyl tartrate of compound B is then converted to its free base by treating the diastereomeric salt with sodium carbonate.

The enantiomeric purity of compound B obtained by the method of this invention was evaluated and compared to that obtained using the re-dissolution method reported in the aforementioned published US patent application No. 2004/0106581 using chiral HPLC. It was proved that compound B obtained by the method of the present invention is one isomer with 100% e.i. (enantiomeric excess), while compound B was obtained with 88.3% e.i., when obtained by the method disclosed in US patent application No. 2004/0106581.

Thus, not only the simple enantiomers obtained by the method according to this invention have greater enantiomeric purity compared to simple enantiomers obtained by known methods, but the method according to this invention is simpler and more economical than the known methods, since it involves one step reactions to produce a diastereomeric salt, namely a dibenzoyl tartrate salt. This method includes a single crystallization. The use of a chiral additive (-) - DBTA has the additional advantage that it is relatively cheaper than (+) - DBTA, which are used in the method described in published patent application US No. 2004/0106581, thereby reducing the total cost of production .

The racemic compound of formula (VIA), key intermediates in the preparation of the compounds of formula (I), can be prepared as set forth in the following Scheme 1.

The production steps according to the compound of formula (V), starting from the compound of formula (II), are described in US-A-4900727, which is incorporated herein by reference. In converting the compound of formula (V) to formula (VIA) in the above scheme, the hydroxyl function of the piperidine ring can be converted to a leaving group, such as tosyl, mesyl, triflate or halide, by treatment with an appropriate reagent such as p-toluenesulfonyl chloride, methanesulfonyl chloride, trifluoromethanesulfonic anhydride or phosphorus pentachloride in the presence of oxygen nucleophiles, such as triethylamine, pyridine, potassium carbonate or sodium carbonate, followed by narrowing of the ring in the presence of oxygen nucleophiles, such as ac sodium acetate or potassium acetate in an alcohol solvent such as isopropanol, ethanol or propanol. The narrowing of the ring included in this step may be performed before the formation of the flavon, as shown in the above diagram, or may be completed after the formation of the flavon with the necessary substituents.

One embodiment of the present invention is an enantiomerically pure (-) - trans enantiomer of a compound of formula (VIA) as defined above, wherein enantiomeric purity is as defined.

Another embodiment of the present invention is the use of an enantiomerically pure (-) - trans enantiomer of an intermediate of a compound of formula (VIA) as defined, to produce an enantiomerically pure compound of formula (1) as defined. Using an intermediate having high enantiomeric purity as the starting compound in the method, the resulting (+) - trans enantiomer of pyrrolidines substituted with flavone, represented by formula (1) obtained by the method has correspondingly high enantiomeric purity.

According to a preferred embodiment of the present invention, there is provided a method for producing an enantiomerically pure (+) - trans enantiomer of a compound of formula (I), or a pharmaceutically acceptable salt thereof, from a dissolved enantiomerically pure (-) - trans enantiomer of an intermediate of a compound of formula (VIA), the method comprises stages in which:

(a) treating the dissolved enantiomerically pure (-) - trans enantiomer of the intermediate of the compound of formula (VIA),

,

,

where R ₂ , R ₃ , R ₄ and R ₉ are, as defined, acetic anhydride in the presence of a Lewis acid catalyst to obtain a dissolved acetylated compound of formula (VIIA),

,

,

where R ₂ , R ₃ and R ₉ are as defined, and R ₄ represents CH ₂ OS (O) CH ₃ ;

(b) reacting the dissolved acetylated compound of formula (VIIA) with an acid of formula R ₄ COOH or an acid chloride of formula R ₁ COCl or an acid anhydride of formula (R ₁ CO) ₂ O or an ester of formula R ₁ COOCH ₃ , where R _{1 is as} defined above in the presence of a base and a solvent to obtain a dissolved compound of formula (VIIIA);

,

,

where R ₁ , R ₂ , R ₃ and R ₉ are as defined above, and R ₄ represents CH ₂ OS (O) CH ₃ ;

(c) treating the dissolved compound of formula (VIIIA) with a base in a suitable solvent to obtain the corresponding dissolved β-diketone compound of formula (IXA);

,

,

where R ₁ , R ₂ , R ₃ and R ₉ are as defined above, and R ₄ represents CH ₂ OS (O) CH ₃ ;

(d) treating the dissolved β-diketone compound of formula (IXA) with an acid, such as hydrochloric acid, to obtain the corresponding cyclic compound of formula (XA),

,

,

where R ₁ , R ₂ , R ₃ and R ₉ are as defined above, R ₄ represents CH ₂ OH;

(e) subjecting the compound of formula (XA) to dealkylation by heating it with a dealkylating agent at a temperature in the range of 120-180 ° C. to obtain the (+) - trans enantiomer of the compound of formula (I) (where R ₂ and R ₃ are hydroxy) and, optionally converting the tested compound to its pharmaceutically acceptable salt.

The Lewis acid catalyst used in step (a) above may be selected from: BF ₃ . Et ₂ O, zinc chloride, aluminum chloride and titanium chloride.

The base used in step (b) may be selected from triethylamine, pyridine and a combination of DCC-DMAP (dicyclohexylcarbodiimide-dimethylaminopyridine).

It will be apparent to those skilled in the art that rearrangement of a compound of formula (VIIIA) into the corresponding β-diketone compound of formula (IXA) is known as Baker-Venkataraman rearrangement (J. Chem. Soc., 1381 (1933) and Curr. Sci., 4,214 (1933 )).

The base used in step (c) may be selected from: lithium hexamethyldisiazides, sodium hexamethyldisiazides, potassium hexamethyldisiazides, sodium hydride and potassium hydride. The most preferred base is lithium hexamethyldisiazide.

The dalkylating agent used in step (e) to dealkylate a compound of formula (IXA) may be selected from: pyridine hydrochloride, boron tribromide, boron trifluoride etherate and aluminum trichloride. The most preferred dealkylating agent is pyridine hydrochloride.

An alternative method of obtaining a compound of formula (I) includes the steps of:

(i) hydrolyzing the dissolved acetylated compound of formula (VIIA) (obtained in step (a) of the method described above) with a base in the presence of a solvent to obtain the corresponding dissolved alcohol of formula (XIA);

,

,

where R ₂ , R ₃ and R ₉ are as defined above;

(ii) treating the dissolved compound of formula XIA with an acid of formula R ₁ COOH, or with an acid chloride of formula R ₁ COCl, or with an acid anhydride of formula (R ₁ CO) ₂ O, or an ester of formula R ₁ COOCH ₃ , where R _{1 is as} defined above, in the presence of a base and a suitable solvent in a nitrogen atmosphere, followed by acid-catalyzed cyclization to produce a compound of formula (XA);

i) subjecting the dissolved compound of formula (XA) to dealkylation by heating it with a dealkylating agent at a temperature in the range of 120-180 ° C. to obtain the dissolved (+) enantiomer of the compound of formula (I) (where R ₂ and R ₃ represent a hydroxyl group) and optionally converting the resulting compound into its pharmaceutically acceptable salt.

In step (i) of an alternative method, the base may be selected from sodium hydroxide and potassium hydroxide and the solvent may be selected from: methanol and ethanol.

In step (ii) of an alternative method, the base can be selected from sodium hydride and potassium hydride and the solvent can be selected from: dimethylformamide, tetrahydrofuran and 1,4-dioxane.

This invention also relates to all dissolved intermediates that are essential for the synthesis of the enantiomerically pure (+) - trans-enantiomer of a compound of general formula (I).

More particularly, enantiomerically pure intermediates that are included in the scope of this invention are compounds of formula (VIA), (VIIA), (VIIIA), (IXA), (XA) and (XIA):

In the compounds of formula (VIA), (VIIA), (VIIIA), (IXA), (XA) and (XIA), the substituents R ₁ , R ₂ , R ₃ , R ₄ and R ₉ are as defined above.

In a specific embodiment of the invention, selective demethylation of a compound of formula (XA) [where R ₁ is phenyl substituted with R ₁₀ and R ₁₁ , where R ₁₀ and R _{11 are} independently represented by a group selected from: halogen, nitro, cyano, C ₁ -C ₄ -alkyl, fluoromethyl, difluoromethyl, trifluoromethyl, C ₁ -C ₄ -alkoxy, hydroxyl, carboxy, C ₁ -C ₄ -alkoxycarbonyl, C ₁ -C ₄ -alkylene hydroxy, CONH ₂ , CONR ₅ R ₆ , SO ₂ NR ₅ R ₆ , cycloalkyl, NR ₅ R ₆ , SR ₇ ; and R ₄ represents CH ₂ OH] carried out as shown in the following scheme 2.

As illustrated in Scheme 2 above, the hydroxy group in the compound of formula (XA) is protected by a suitable protecting group, such as an acetyl group, by treating said compound of formula (XA) with an appropriate reagent, such as acetyl chloride or acetic anhydride, in the presence of a base selected from: sodium hydride potassium hydride, pyridine, triethylamine and dimethylaminopyridine (DMAP) to obtain an acetylated compound of formula XIIA. A compound of formula XIIA is then treated with a demethylating agent, such as boron trifluoride etherate, to give a compound of formula XIIIA.

The present invention encompasses prodrugs of enantiomerically pure compounds of formula (I) and pharmaceutical compositions containing them and a method of treatment by administering such prodrugs.

Another aspect of the present invention provides a process for preparing a prodrug of a dissolved compound of formula (I), the process comprising the steps of treating a dissolved compound of formula (I), wherein R ₂ and R ₃ are hydroxyl, acid or acid chloride in the presence of a base to produce a prodrug of a dissolved compound formula (1), where R ₂ represents hydroxy, R ₃ represents hydroxy, OCOR or OR, and R ₄ represents alkylene OH or alkylene OS (O) R; where R represents alkyl or aryl. One such prodrug is a compound of formula (XIVA).

An example of obtaining a prodrug of an enantiomerically pure compound of formula (I) is depicted in the following scheme 3

A method comprising reacting an enantiomerically pure compound of formula (I), wherein R ₂ and R ₃ are hydroxy, with an acid or acid chloride in the presence of a base selected from: sodium hydride, potassium hydride, pyridine, trimethylamine and DMAP to obtain enantiomerically pure ( +) - the trans-enantiomer of a prodrug of formula (XIVA), where R ₂ is hydroxy,

R ₃ is OC (O) R, R ₄ is alkylene O (O) R; where R represents alkyl or aryl. In the compounds of formula XIVA and I, R ₁₀ and R ₁₁ are as defined above.

According to this invention, the compounds of general formula (I) are inhibitors of CDK, in particular CDK / cyclin complexes and are used in antiproliferative therapies for diseases that are characterized by excessive cell growth, such as cancer, cardiovascular disorders, nephrological disorders, psoriasis, parasitology, Alzheimer's disease, inflammation and arthritis, immunological disorders, including unwanted proliferation of leukocytes, restenosis and other proliferative disorders of the smooth muscle, viral infections and fungal infections.

The present invention thus relates to enantiomerically pure compounds of formula (I), and / or their pharmaceutically acceptable salts, and / or their prodrugs for use as medicaments (or medicines), for use of enantiomerically pure compounds of formula (I), and / or their pharmacologically acceptable salts and / or their prodrugs for the manufacture of medicaments for inhibiting cell proliferation or for the treatment and prophylaxis of the diseases mentioned above, for example, for the manufacture of medicaments for those FIPA and prevention of cancer, inflammation and arthritis, psoriasis, bone diseases, mycotic or viral infections, cardiovascular disease, Alzheimer's disease, etc., and to methods of treatment aiming at such purposes including methods for said therapies and prophylaxis. The invention further relates to pharmaceutical compositions which contain an effective amount of at least one enantiomerically pure compound of formula (I) and / or its physiologically acceptable salts and / or its prodrugs, in addition to a conventional pharmaceutically acceptable carrier, optionally, with other active antiproliferative agents, and to a process for the preparation of a medicament which comprises introducing at least one enantiomerically pure compound of formula (I) in a suitable administration form, using a pharmaceutically acceptable and physiologically suitable carrier and, if appropriate, additionally suitable active compounds and / or additives. These compositions can be prepared using known techniques in the art, such as those described in Remington's Pharmaceutical Sciences, published by Mack Publishing Co. or "Pharmaceutical Dosage Form and Drug Delivery Systems" (sixth edition) published by Williams & Wilkins, (1995), each incorporated by reference in this description.

This invention also relates to a method for producing a medicament for the treatment or prevention of diseases associated with excessive cell proliferation, which is characterized in that at least one enantiomerically pure compound of the general formula (I) is used as a pharmaceutically active substance.

Pharmaceutical compositions typically contain from about 1 to 99%, preferably from about 5 to 70%, most preferably from about 10 to about 30% by weight of enantiomerically pure compounds of formula (I) and / or from physiologically suitable salts and / or their prodrugs. The amount of the active ingredient of the enantiomerically pure compound of formula (I) and / or its physiologically suitable salts and / or its prodrugs in pharmaceutical compositions is usually about 5 to 500 mg. The dosage of the enantiomerically pure compounds of this invention that are administered may span a wide range. The dose to be administered daily is selected to satisfy the desired effect. About 20 to 1000 mg are preferably administered daily to the patient. If necessary, you can also enter a higher or lower dose. Actual dose levels of the active ingredients in the pharmaceutical compositions of this invention may vary so as to obtain an amount of the active ingredient that is effective to achieve the desired therapeutic response for a particular patient, composition and administration without poisoning the patient.

The dose level chosen will depend on various factors, including the activity of the particular compound of this invention or of the ester, salt or amide thereof, route of administration, time of administration, rate of excretion of the particular compound used, duration of treatment, other drugs, compounds and / or materials used in combination with the individual compounds used, the patient’s age, gender, weight, condition, general health, and preliminary medical history of treated, and like factors well known in medicine.

Medications can be administered orally, including sublingually, for example in the form of pills, tablets, coated tablets, capsules, granules, powders, syrups or elixirs. The introduction, however, can also be carried out: rectally or vaginally, for example in the form of suppositories; parenterally, for example, intravenously, intramuscularly or subcutaneously, including the use of infusion techniques in the form of injectable sterile solutions or suspensions; topically, for example, in the form of creams, ointments, lotions, foams, gels, emulsions, solutions, tinctures, chatters or transdermal patches; or in other ways, for example ophthalmically, optically, nasally or in other forms, for example aerosols, inhalers or nasal sprays.

The pharmaceutical compositions of this invention are prepared by a method known per se and known to one skilled in the art. Pharmaceutically acceptable inert inorganic and / or organic carriers and / or additives can be used in addition to the compound (s) of formula (I) and / or its (their) physiologically suitable salts and / or its (their) prodrugs. To obtain pills, tablets, coated tablets and hard gelatin capsules, it is possible to use, for example, lactose, corn starch or its derivatives, gum arabic, magnesia or glucose, etc. Carriers for soft gelatin capsules and suppositories are, for example, fats, waxes, natural or hydrogenated oils, etc. Suitable carriers for the manufacture of solutions are, for example, injection solutions, or emulsions, or syrups, for example water, physiological saline, or alcohols, for example ethanol, propanol or glycerol, sugar solutions, such as glucose solutions or mannitol solutions, or mixtures of various solvents that were mentioned. Additives that can be used are, for example, fillers, antioxidants, dispersants, emulsifiers, anti-foaming agents, aroma modifiers, preservatives, solubilizers or colorants.

Commonly used pharmaceutically acceptable carriers and additives that can be used as intended to formulate a composition for its intended route of administration are well known in the art and include those carriers and additives described in US Patent Application Laid-Open No. 2004/0106581.

The pharmaceutical compositions may also contain two or more enantiomerically pure compounds of the formula (I), and / or their physiologically suitable salts, and / or their prodrugs. Moreover, in addition to at least one enantiomerically pure compound of formula (I) and / or its physiologically suitable salts and / or its prodrugs, the pharmaceutical compositions may also contain one or more other therapeutically or prophylactically active ingredients. Thus, the compounds of this invention can be used as drugs in the treatment of proliferative disorders, either singly or as part of combination therapies. For example, enantiomerically pure compounds of the present invention can be used in combination with known anti-cancer, cytostatic and cytotoxic agents. If formulated as a fixed dose, such combination products use the compounds of this invention within the dosage range described above, and another pharmaceutically active agent within their permitted dosage range. For example, it has been found that the CDK inhibitor olomucin acts synergistically with known cytotoxic agents in inducing apoptosis (J. Cell Sci., 1995, 108, 2897). Enantiomerically pure compounds of the general formula (I) can be used sequentially with known drugs, such as anti-cancer or cytotoxic agents, when the combination is not suitable.

Optional antiproliferative agents that can be added to the composition include, but are not limited to, compounds that are described in cancer chemotherapy drug regimens in the 13th edition of the Merck Index, (1996) (NB or corrected), which are described herein incorporated by reference, such as asparaginase, bleomycin, carboplatin, carmustine, chlorambucil, cisplatin, colaspase, cyclophosphamide, cytarabine, dacarbazine, dactinomycin, daunorubicin, doxorubicin, epirubicin, etoposylamine, hydroxymera, fluorouracil y, ifosfamide, leucovorin, lomustine, mechlorethamine, 6-mercaptopurine, mesna, methotrexate, mitomycin C, mitoxantrone, prednisolone, prednisone, procarbazine, streptozocin, tamoxifen, thioguanine, vinblastine, vincincinine.

Other antiproliferative agents suitable for use with the composition of the invention include, but are not limited to, those recognized for use in the treatment of neoplasms in Goodman and Gilman's The Pharmacological Basis of Therapeutics (ninth edition), editor Molinoff et al., Publ. McGraw-Hill, pp. 1225-1287, (1996), which are incorporated herein by reference, such as aminoglutethimide, L-asparaginase, azathioprine, 5-azacytidine, cladribine, bisulfan, diethylstilbestrol, 2 ', 2'-difluorodeoxycytidine, docetaxel , eritrogidroksinoniladenin, ethinyl estradiol, 5-fluorodeoxyuridine, 5-fluorodeoxyuridine monophosphate, fludarabinfosfat, fluoxymestrone, flutamide, gidroksiprogesteronkaproat, idarubicin, interferon, medroksipogesteronatsetat, megestrol acetate, melphalan, mitotane, paclitaxel, pentostatin, N-fosfonoatsetil-L-aspartate (PALA), plicamycin , this Springsteen, teniposide, testosterone propionate, thiotepa, trimetilmelamin, uridine, vinorelbine.

Optional cyclin-dependent kinase (CDK) inhibitory agents that can be added to the composition include, but are not limited to, alsterpaullon, butyrolactone I, CDK2 inhibitor, CDK2 / cyclin inhibitory peptide I, CDK2 / cyclin inhibitory peptide II, 2- (2- hydroxyethylamino) -6- (3-chloroaniline) -9-isopropylpurine, indirubin-3'-monoxime, kenpaullon, olomucin, isolomucin, N ⁹ -isopropyl-olomucin, purvalanol A, roskovitin, the (S) -isomer of rosquitin and WHI-P180 [4- (3'-hydroxyphenyl) amino-6,7-dimethoxyquinazoline] (from Calbiochem Signal Transduction Catalog & Technical Resource 2001).

It is understood that modifications that do not significantly affect the activity of various embodiments of the invention are included within the scope of the disclosed invention. Accordingly, the following examples illustrate but do not limit the invention.

Examples

The following abbreviations are used in this description:

BF ₃ : Boron trifluoride diethyl ether

Conc .: Concentrated

THF: Tetrahydrofuran

EtOAc: Ethyl Acetate

MeOH: Methanol

IPA: Isopropanol

DBTA: Dibenzoyl tartaric acid

DMF: N, N-dimethylformamide

DCC: N, N-Dicyclohexylcarbodiimide

DMAP: (N, N-dimethylamino) pyridine

HCl: Hydrochloric acid

NaBH ₄ : Sodium borohydride

NaOH: Sodium Hydroxide

Na ₂ CO ₃ : Sodium Carbonate

Example 1:

1-Methyl-4- (2,4,6-trimethoxyphenyl) -1,2,3,6-tetrahydropyridine

1-methyl-4-piperidone (340 g, 3.0 mol) was slowly added to a solution of 1,3,5-trimethoxybenzene (500 g, 2.97 mol) in glacial acetic acid (600 ml), keeping the temperature of the reaction mixture below 40 ° FROM. Conc. HCl (450 ml) was added after 20 minutes. The temperature was raised to 85-90 ° C and the reaction mixture was stirred for 3.5 hours. Cooled to 40 ° C, poured onto crushed ice (4 kg) and stirred for 20 minutes. The precipitate of unreacted 1,3,5-trimethoxybenzene was filtered. The filtrate was made alkaline below 10 ° C to pH 11-12 using a 50% aqueous NaOH solution. The resulting whitish solid was filtered, washed with water and dried to obtain 1-methyl-4- (2,4,6-trimethoxyphenyl) -1,2,3,6-tetrahydropyridine compound.

Yield: 580 g (74%);

mp (melting point): 112-114 ° C;

IR (infrared radiation) (KBr): 3045, 2900, 2837, 1600, 1585 cm ^-1 ;

¹ H NMR (CDCl ₃ , 300 MHz): δ 6.15 (s, 2H), 5.55 (s, 1H), 3.85 (s, 3H), 3.75 (s, 6H), 3, 10 (d, 2H), 2.55 (t, 2H), 2.40 (s, 3H), 2.35 (m, 2H);

MS (EI): m / z 263 (M +).

Example 2:

(±) -trans-1-methyl-4- (2,4,6-trimethoxyphenyl) piperidin-3-ol

Boron trifluoride diethyl ether (300 ml, 2.36 mol) was slowly added with stirring in a nitrogen atmosphere at 0 ° C to a solution of the compound of formula (1) (300 g, 1.14 mol) and NaBH ₄ (75 g, 1.97 mol) in dry THF (2.25 L). The temperature of the reaction mixture was slowly raised to 55 ° C and stirred for 1.5 hours. It was cooled to 30 ° C. Ice water (100 ml) was slowly added, followed by acidification of conc. HCl (375 ml). The reaction mixture was stirred for 1 hour at 50-55 ° C. It was cooled to 30 ° C and alkalinized using a 50% aqueous solution of NaOH to pH 11-12. Hydrogen peroxide (30%, 225 ml) was added after 0.5 hours. The reaction mixture was stirred at 55-60 ° C for 1.5 hours. It was cooled to 30 ° C and sufficient water was added to dissolve the precipitated salts. The organic layer was separated and the aqueous portion was extracted with ethyl acetate (2 × 1 L). The organic extracts were dried (anhydrous Na ₂ SO ₄ ) and concentrated. The resulting crude viscous brown oil was treated with 4N HCl (1.2 L) and extracted with ethyl acetate (2 × 500 ml). The aqueous portion was cooled, alkalinized with a 50% aqueous sodium hydroxide solution and extracted using ethyl acetate (2 × 1 L). The organic extract was dried (anhydrous Na ₂ SO ₄ ) and concentrated to obtain the compound (±) -trans-1-methyl-4- (2,4,6-trimethoxyphenyl) -piperidin-3-ol.

Yield: 210 g (65.6%);

mp: 96-97 ° C;

IR (KBr): 3582, 3374, 3017 cm ^-1 ;

¹ H NMR (CDCl ₃ , 300 MHz): δ 6.15 (s, 2H), 4.40 (m, 1H), 3.79 (s, 3H), 3.74 (s, 6H), 3, 20 (dd, 1H), 3.00 (m, 1H), 2.80 (m, 1H), 2.40 (m, 1H), 2.37 (s, 3H), 2.00 (m, 1H ), 1.90 (t, 1H), 1.52 (m, 1H);

MS (CI): m / z 282 (M + 1).

Example 3:

(±) -trans-acetic acid-1-methyl-3- (2,4,6-trimethoxyphenyl) pyrrolidin-2-yl methyl ester

Methanesulfonyl chloride (30.27 ml, 44.8 g, 0.4 mol) was added dropwise to a cooled and stirred solution of the compound of formula (2) (100 g, 0.35 mol) and triethylamine (71.88 g, 0, 7 mol) in dry THF (1.0 L). The reaction mixture was further stirred for 45 minutes at 0 ° C. The precipitate of triethylamine HCl was filtered and washed with dry THF (2 × 100 ml). The filtrate was added dropwise to a suspension of sodium acetate, refluxing (115 g, 1.40 mol) in 2-propanol (1.0 L). The reaction mixture was refluxed for an additional 15 minutes, diluted with EtOAc (1.0 L) and the salts were filtered. The mixture of salts was washed with EtOAc (2 × 100 ml). The combined filtrate was concentrated to obtain gum. Water (50 ml) was added to the gum with stirring to obtain a solid, which was filtered and dried to obtain the compound, (±) -trans-acetic acid 1-methyl-3- (2,4,6-trimethoxyphenyl) -pyrrolidin-2 -yl methyl ester.

Yield: 90 g (81%);

mp: 74-77 ° C;

¹ H NMR (CDCl ₃ , 300 MHz): δ 6.13 (s, 2H), 4.00 (m, 2H), 3.81 (m, 1H), 3.79 (s, 3H), 3, 76 (s, 6H), 3.20 (m, 1H), 2.75 (m, 1H), 2.69 (m, 1H), 2.47 (s, 3H), 2.00 (m, 2H) ), 1.99 (s, 3H).

Example 4:

(±) -trans- [1-Methyl-3- (2,4,6-trimethoxyphenyl) pyrrolidin-2-yl] methanol

A 10% aqueous solution of NaOH (596 ml) was added to a solution of the compound of Example (3) (241 g, 0.75 mol) in methanol (596 ml). The reaction mixture was stirred at 50 ° C for 45 minutes. It was concentrated to gum and then poured into ice water (2 L). The resulting solid was filtered to obtain the compound (±) -trans- [1-methyl-3- (2,4,6-trimethoxyphenyl) pyrrolidin-2-yl] methanol.

Yield: 198 g (94%);

mp: 82-85 ° C;

IR (KBr): 3421, 3009, 1607 cm ^-1 ;

¹ H NMR (CDCl ₃ , 300 MHz): δ 6.15 (s, 2H), 3.92 (m, 1H), 3.80 (s, 9H), 3.60 (dd, 1H), 3, 45 (d, 1H), 3.20 (m, 1H), 2.78 (m, 1H), 2.50 (m, 1H), 2.42 (s, 3H), 2.00 (m, 1H ), 1.92 (m, 1H);

MS (ES +): m / z 282 (M + 1).

Example 5:

(-) - trans- [1-Methyl-3- (2,4,6-trimethoxyphenyl) pyrrolidin-2-yl] methanol

(-) - DBTA (321.7 g, 897.7 mmol) was added to the compound of formula (4) (250 g, 889.6 mmol), followed by the addition of methanol (1715 ml). The mixture was heated under reflux for 10 minutes, slowly stirred at room temperature for 3 hours, the crystallized salt was filtered and dried.

Yield: 185 g (30%);

mp: 102-105 ° C;

[α] _D ²⁵ = -82.66 ° (c = 07, methanol).

The salt was mixed with a 10% aqueous solution of Na ₂ CO ₃ (765 ml) and EtOAc (200 × 3 ml) to obtain the free base in an EtOAc layer. The EtOAc layer was concentrated to obtain the compound (-) - trans- [1-methyl-3- (2,4,6-trimethoxyphenyl) pyrrolidin-2-yl] methanol.

Yield: 80 g (98.3%);

[α] _D ²⁵ = -20.0 ° (c = 0.7, methanol);

¹ H NMR (CDCl ₃ , 300 MHz): δ 6.13 (s, 2H), 3.90 (m, 1H), 3.79 (s, 9H), 3.57 (dd, 1H), 3, 38 (d, 1H), 3.13 (m, 1H), 2.69 (m, 1H), 2.47 (m, 1H), 2.34 (s, 3H), 2.00 (m, 1H ), 1.93 (m, 1H).

This compound was subjected to chiral HPLC. Chiral HPLC was performed using a Chiralcel OD-H column (250 × 4.6 mm) and a hexane: ethanol solvent system (92:08) with TFA (0.4%). The results were recorded at 264 nm with a solvent flow rate of 1 ml / min. As depicted in figure 1, chiral HPLC shows 100% e.i. compounds, (-) - trans- [1-Methyl-3- (2,4,6-trimethoxyphenyl) pyrrolidin-2-yl] methanol.

Example 6:

(-) - trans-acetic acid-3- (3-acetyl-2-hydroxy-4,6-dimethoxyphenyl) -1-methylpyrrolidin-2-yl methyl ester

BF ₃ etherate (25.2 g, 178 mmol) was added dropwise with stirring at 0 ° C, in an N ₂ atmosphere, to a solution of the compound of example (5) (10 g, 35.58 mmol) in acetic anhydride (19.48 ml, 176 mmol). The reaction mixture was stirred at room temperature for 2 hours. It was poured onto crushed ice (1 kg), alkalized using a saturated aqueous solution of Na ₂ CO ₃ and extracted using EtOAc (3 × 200 ml). The organic extract was washed with brine, dried (anhydrous Na ₂ SO ₄ ) and concentrated to give 3- (3-acetyl-2-hydroxy-4,6-dimethoxy-phenyl) -1-methyl-pyrrolidine- (-) -trans-acetic acid compound 2-yl methyl ester.

Yield: 10 g (83%);

¹ H NMR (CDCl ₃ , 300 MHz): δ 14.20 (s, 1H), 5.96 (s, 1H), 4.10 (d, 2H), 3.90 (s, 3H), 3, 89 (s, 3H), 3.85 (m, 1H), 3.26 (m, 1H), 2.82 (m, 1H), 2.74 (m, 1H), 2.66 (s, 3H ), 2.52 (s, 3H), 2.21 (m, 2H), 2.10 (s, 3H).

Example 7

(-) - trans-1- [2-Hydroxy-3- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -4,6-dimethoxyphenyl) ethanone

To a solution of the compound of Example (6) (10 g, 28.4 mmol) in methanol (25 ml) was added with stirring at room temperature a 10% aqueous solution of NaOH (25 ml). The temperature of the reaction mixture was raised to 50 ° C in 45 minutes. It was cooled to room temperature, acidified using conc. HCl, and concentrated to remove methanol. It was alkalinized using a saturated aqueous solution of Na ₂ CO ₃ . The compound (-) - trans-1- [2-hydroxy-3- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -4,6-dimethoxyphenyl) ethanone was filtered off, washed with water and dried.

Yield: 7.14 g (82%);

IR (KBr): 3400, 3121, 3001, 1629, 1590 cm ^-1 ;

¹ H NMR (CDCl ₃ , 300 MHz): δ 5.96 (s, 1H), 3.93 (m, 1H), 3.90 (s 3H), 3.88 (s, 3H), 3.59 (dd, 1H), 3.37 (d, 1H), 3.13 (m, 1H), 2.75 (m, 1H), 2.61 (s, 3H), 2.59 (m, 1H) 2.37 (s, 3H); 2.00 (m, 2H);

MS (ES +): m / z 310 (M + 1).

Example 8:

(+) - trans-2- (2-Chlorophenyl) -8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -5,7-dimethoxychromen-4-one

Sodium hydride (50%, 0.54 g, 11.25 mmol) was added in portions to a solution of the compound of example (7) (0.7 g, 2.2 mmol) in dry DMF (15 ml) at 0 ° C. in atmosphere nitrogen and with stirring. After 10 minutes, methyl 2-chlorobenzoate (1.15 g, 6.75 mmol) was added. The reaction mixture was stirred at 25 ° C for 2 hours. Methanol was carefully added at below 20 ° C. The reaction mixture was poured onto crushed ice (300 g), acidified with 1: 1 HCl (pH 2) and extracted using EtOAc (2 × 100 ml). The aqueous layer was alkalinized using saturated Na ₂ CO ₃ (pH 10) and extracted using CHCl ₃ (3 × 200 ml). The organic layer was dried (anhydrous Na ₂ SO ₄ ) and concentrated. To the residue was added conc. HCl (25 ml) and stirred at room temperature for 2 hours. The reaction mixture was poured onto crushed ice (300 g) and made basic using a saturated aqueous solution of Na ₂ CO ₃ . The mixture was extracted using CHCl ₃ (3 × 200 ml). The organic extract was washed with water, dried (anhydrous Na ₂ SO ₄ ) and concentrated to obtain the compound (+) - trans-2- (2-chlorophenyl) -8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -5, 7-dimethoxychromen-4-one.

Yield: 0.67 g (64%);

mp: 91-93 ° C;

[α] _D ²⁵ = + 5.8 ° (c = 0.7, methanol);

IR (KBr): 3431, 1648, 1598, 1571 cm ^-1 ;

¹ H NMR (CDCl ₃ , 300 MHz): δ 7.70 (dd, 1H), 7.52 (m, 1H), 7.45 (m, 2H), 6.50 (s, 1H), 6, 44 (s, 1H), 4.17 (m, 1H), 4.00 (s, 3H), 3.97 (s, 3H), 3.64 (dd, 1H), 3.40 (d, 1H) ), 3.15 (m, 1H), 2.74 (d, 1H), 2.52 (m, 1H), 2.32 (s, 3H), 2.00 (m, 2H);

MS (ES +): m / z 430 (M + 1).

Example 9:

(+) - trans-2- (2-Chlorophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one

Molten pyridine hydrochloride (4.1 g, 35.6 mmol) was added to the compound of formula (8) (0.4 g, 0.9 mmol) and heated at 180 ° C. for 1.5 hours. The reaction mixture was cooled to 25 ° C., diluted with MeOH (10 ml) and made basic using Na ₂ CO ₃ to pH 10. The mixture was filtered and the organic layer was concentrated. The residue was suspended in water (5 ml), stirred for 30 minutes, filtered and dried to obtain compound (+) - trans-2- (2-chlorophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1- methylpyrrolidin-3-yl) -chromen-4-one.

Yield: 0.25 g (70%);

IR (KBr): 3422, 3135, 1664, 1623, 1559 cm ^-1 ;

¹ H NMR (CDCl ₃ , 300 MHz): δ 7.56 (d, 1H), 7.36 (m, 3H), 6.36 (s, 1H), 6.20 (s, 1H), 4, 02 (m, 1H), 3.70 (m, 2H), 3.15 (m, 2H), 2.88 (m, 1H), 2.58 (s, 3H), 2.35 (m, 1H ), 1.88 (m, 1H);

MS (ES +): m / z 402 (M + 1);

Study: C ₂₁ H ₂₀ ClNO ₅ C, 62.24 (62.71); H, 5.07 (4.97); N, 3.60 (3.48); Cl, 9.01 (8.83).

Example 10:

(+) - trans-2- (2-chlorophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one hydrochloride

The compound of formula (9) (0.2 g, 0.48 mmol) was suspended in IPA (isopropyl alcohol) (5 ml) and 3.5% HCl (25 ml) was added. The suspension was heated until a clear solution was obtained. The solution was cooled and the solid was filtered to obtain the compound (+) - trans-2- (2-Chlorophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one hydrochloride.

Yield: 0.21 g (97%);

mp: 188-192 ° C;

[α] _D ²⁵ = + 21.3 ° (c = 0.2, methanol);

¹ H NMR (CD ₃ OD, 300 MHz): δ 7.80 (d, 1H), 7.60 (m, 3H), 6.53 (s, 1H), 6.37 (s, 1H), 4 23 (m, 1H), 3.89 (m, 2H), 3.63 (m, 1H), 3.59 (dd, 1H), 3.38 (m, 1H), 2.90 (s, 3H); 2.45 (m, 1H); 2.35 (m, 1H);

MS (ES +): m / z 402 (M + 1) (free base).

This compound was subjected to chiral HPLC. Chiral HPLC was performed using a Chiralcel OD-H column (250 × 4.6 mm) and a hexane: ethanol solvent system (92:08) with TFA (0.4%). The results were recorded at 264 nm with a solvent flow rate of 1 ml / min. As depicted in FIG. 3, chiral HPLC shows 100% e.i. compounds, (+) - trans-2- (2-chlorophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one hydrochloride.

Example 11:

(+) - trans-2- (2-Chloro-5-fluorophenyl) -8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -5,7-dimethoxychromen-4-one

Sodium hydride (50%, 0.776 g, 16 mmol) was added in portions to a solution of the compound of formula (7) (1.0 g, 3.2 mmol) in dry DMF (25 ml) at 0 ° C., under nitrogen and by stirring. After 10 minutes, methyl 2-chloro-5-fluorobenzoate (1.22 g, 6.4 mmol) was added. The reaction mixture was stirred at 25 ° C for 2 hours. Methanol was carefully added at below 20 ° C. The reaction mixture was poured onto crushed ice (300 g), acidified with 1: 1 HCl (pH 2) and extracted using EtOAc (2 × 100 ml). The aqueous layer was alkalinized using saturated Na ₂ CO ₃ (pH 10) and extracted using CHCl ₃ (3 × 200 ml). The organic layer was dried (anhydrous Na ₂ SO ₄ ) and concentrated. To the residue was added conc. HCl (25 ml) and stirred at room temperature for 2 hours. The reaction mixture was poured onto crushed ice (300 g) and made basic using a saturated aqueous solution of Na ₂ CO ₃ . The mixture was extracted using CHCl ₃ (3 × 200 ml). The organic extract was washed with water, dried (anhydrous Na ₂ SO ₄ ) and concentrated to obtain the compound (+) - trans-2- (2-chloro-5-fluorophenyl) -8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl ) -5,7-dimethoxychromen-4-one.

Yield: 0.9 g (63%);

¹ H NMR (CDCl ₃ , 300 MHz): δ 7.57 (m, 1H), 7.46 (m, 1H), 7.16 (m, 1H), 6.58 (s, 1H), 6, 45 (s 1H), 4.15 (m, 1H), 4.00 (s, 3H), 3.97 (s, 3H), 3.63 (dd, 1H), 3.32 (d, 1H) 3.13 (m, 1H); 2.61 (m, 1H); 2.53 (m, 1H); 2.29 (s, 3H); 2.00 (m, 2H); MS (CI):

m / z 448 (M + 1).

Example 12:

(+) - trans-2- (2-Chloro-5-fluorophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one

Molten pyridine hydrochloride (8.0 g, 69.5 mmol) was added to the compound of formula (11) (0.8 g, 1.78 mmol) and heated at 180 ° C. for 1.5 hours. The reaction mixture was cooled to 25 ° C., diluted with MeOH (10 ml) and made basic using Na ₂ CO ₃ to pH 10. The mixture was filtered and the organic layer was concentrated. The residue was suspended in water (10 ml), stirred for 30 minutes, filtered and dried to give compound (+) - trans-2- (2-chloro-5-fluorophenyl) -5,7-dihydroxy-8- (2- hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one.

Yield: 0.45 g (60%);

mp: 253-254 ° C;

IR (KBr): 3450, 1665 cm ^-1 ;

¹ H NMR (DMSO d ₆ , 300 MHz): δ 12.70 (s, 1H), 7.75 (m, 2H), 7.54 (m, 1H), 6.55 (s, 1H), 6 14 (s, 1H), 3.80 (m, 1H), 3.51 (m, 3H), 2.94 (m, 2H), 2.46 (s, 3H), 2.15 (m, 1H), 1.86 (m, 1H);

MS (ES +): m / z 420 (M + 1);

Study: C ₂₁ H ₁₉ ClFNO ₅ C, 60.2 (60.08); H, 4.53 (4.56); N, 3.86 (3.34); Cl, 8.17 (8.44).

Example 13:

(+) - trans-2- (2-Chloro-5-fluorophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one hydrochloride

The compound of formula (12) (0.1 g, 0.244 mmol) was suspended in IPA (5 ml) and 3.5% HCl (25 ml) was added. The suspension was heated until a clear solution was obtained. The solution was cooled and the solid was filtered to obtain the compound (+) - trans-2- (2-chloro-5-fluorophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chrome -4-she hydrochloride.

Yield: 0.108 g (100%);

[α] _D ²⁵ = + 18.05 ° (c = 0.7, methanol);

¹ H NMR (CD ₃ OD, 300 MHz): δ 7.67 (m, 2H), 7.37 (m, 1H), 6.60 (s, 1H), 6.37 (s, 1H), 4 25 (m, 1H), 3.90 (m, 2H), 3.69 (m, 1H), 3.60 (dd, 1H), 3.45 (m, 1H), 2.98 (s, 3H); 2.52 (m, 1H); 2.29 (m, 1H);

MS (ES +): m / z 420 (M-36.5).

Example 14:

(+) - trans-2- (2-Bromo-5-fluorophenyl) -8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -5,7-dimethoxychromen-4-one

The compound of formula (7) (6 g, 19.42 mmol) in dry DMF (60 ml) was reacted with methyl 2-bromo-5-fluorobenzoate (6.7 g, 28.75 mmol) in the presence of NaH (50%, 3 88 g, 80.8 mmol) at 0 ° C in a nitrogen atmosphere. The reaction mixture was stirred at 25 ° C for 2 hours. Methanol was carefully added at below 20 ° C.

The reaction mixture was poured onto crushed ice (300 g), acidified with 1: 1 HCl (pH 2) and extracted using EtOAc (2 × 100 ml). The aqueous layer was alkalinized using saturated Na ₂ CO ₃ (pH 10) and extracted using CHCl ₃ (3 × 200 ml). The organic layer was dried (anhydrous Na ₂ SO ₄ ) and concentrated. To the residue was added conc. HCl (25 ml) and stirred at room temperature for 2 hours. The reaction mixture was poured onto crushed ice (300 g) and made basic using a saturated aqueous solution of Na ₂ CO ₃ . The mixture was extracted using CHCl ₃ (3 × 200 ml). The organic extract was washed with water, dried (anhydrous Na ₂ SO ₄ ) and concentrated to obtain the compound (+) - trans-2- (2-bromo-5-fluorophenyl) -8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl ) -5,7-dimethoxychromen-4-one.

Yield: 3.94 g (41.2%);

¹ H NMR (CDCl ₃ , 300 MHz): δ 7.65 (m, 1H), 7.45 (m, 1H), 7.10 (m, 1H), 6.47 (s, 1H), 6.45 (s, 1H), 4.15 (m , 1H), 4.00 (s, 3H), 3.97 (s, 3H), 3.64 (dd, 1H), 3.35 (d, 1H), 3.10 (m, 1H), 2.64 (m, 1H), 2.45 (m, 1H), 2.27 (s, 3H), 2.00 (m, 2H); MS (ES +): m / z 493 (M + 1).

Example 15:

(+) - trans-2- (2-Bromo-5-fluorophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one

Molten pyridine hydrochloride (39 g, 339 mmol) was added to the compound of formula (14) (3.9 g, 7.92 mmol) and heated at 180 ° C. for 1.5 hours. The reaction mixture was cooled to 25 ° C., diluted with MeOH (10 ml) and made basic using Na ₂ CO ₃ to pH 10. The mixture was filtered and the organic layer was concentrated. The residue was suspended in water (40 ml), stirred for 30 minutes, filtered and dried to obtain the compound (+) - trans-2- (2-bromo-5-fluorophenyl) -5,7-dihydroxy-8- (2- hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one.

Yield: 1.4 g (38.14%);

mp: 145-147 ° C;

IR (KBr): 3650, 1640 cm ^-1 ;

¹ H NMR (CDCl ₃ + TFA, 300 MHz): δ 12.40 (s, 1H), 7.55 (m, 1H), 7.28 (m, 1H), 7.00 (m, 1H), 6.31 (s, 1H), 6.28 (s, 1H), 3.98 (m, 1H), 3.68 (m, 2H), 3.50 (m, 2H), 3.15 (m , 1H), 2.80 (s, 3H), 2.30 (m, 1H), 2.08 (m, 1H);

MS (ES +): m / z 465 (M + 1).

Example 16:

(+) - trans-2- (2-Bromo-5-fluorophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one hydrochloride

The compound of formula (15) (1.0 g) was converted to its hydrochloride salt by suspension in IPA (5 ml) and 3.5% HCl (25 ml). The suspension was heated until a clear solution was obtained. The solution was cooled and the solid was filtered to obtain the compound (+) - trans-2- (2-bromo-5-fluorophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chrome -4-she hydrochloride.

Yield: 1.0 g (93%); MS (ES +): m / z 465 (M + 1) (free base).

Example 17:

(+) - trans-2- (2-Bromo-5-fluorophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one methanesulfonate

The compound of formula (15) (1.97 g, 4.25 mmol) in methanol (5 ml) was treated with a solution of methanesulfonic acid in methanol (0.408 g, 4.25 mmol). The resulting clear solution was concentrated to obtain the title compound (+) - trans-2- (2-bromo-5-fluorophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chrome- 4-methanesulfonate.

Yield: 2.3 g (96.63%);

¹ H NMR (D ₂ O, 300 MHz): δ 7.58 (m, 1H), 7.20 (m, 1H), 7.11 (m, 1H), 6.30 (s, 1H), 6 02 (s, 1H), 4.02 (m, 1H), 3.85 (m, 2H), 3.45 (m, 2H), 3.35 (m, 1H), 2.83 (s, 3H), 2.70 (s, 3H), 2.20 (m, 2H);

Study: C ₂₂ H ₂₃ BrNFO ₈ S · H ₂ O, C, 46.08 (45.68); H, 4.61 (4.35); N, 2.63 (2.42), Br, 14.73 (13.81); S, 4.99 (5.54);

MS (ES +): m / z 465 (M + 1), free base.

Example 18:

(+) - trans-2- (2,4-Dichloro-5-fluorophenyl) -8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -5,7-dimethoxychromen-4-one

The compound of formula (7) (0.8 g, 2.58 mmol) in dry DMF (10 ml) was reacted with 2,4-dichloro-5-fluoro-benzoyl chloride (0.887 g, 3.9 mmol) in the presence of NaH (50 %, 0.62 g, 12.9 mmol) at 0 ° C in a nitrogen atmosphere. The reaction mixture was stirred at 25 ° C for 2 hours. Methanol was carefully added at below 20 ° C. The reaction mixture was poured onto crushed ice (100 g), acidified with 1: 1 HCl (pH 2) and extracted using EtOAc (2 × 50 ml). The aqueous layer was made alkaline using saturated Na ₂ CO ₃ (pH 10) and extracted using CHCl ₃ (3 × 100 ml). The organic layer was dried (anhydrous Na ₂ SO ₄ ) and concentrated. To the residue was added conc. HCl (10 ml) and stirred at room temperature for 2 hours. The reaction mixture was poured onto crushed ice (100 g) and made basic using a saturated aqueous solution of Na ₂ CO ₃ . The mixture was extracted using CHCl ₃ (3 × 100 ml). The organic extract was washed with water, dried (anhydrous Na ₂ SO ₄ ) and concentrated to obtain the title compound (+) - trans-2- (2,4-dichloro-5-fluorophenyl) -8- (2-hydroxymethyl-1-methylpyrrolidine- 3-yl) -5,7-dimethoxychromen-4-one.

Yield: 0.54 g (43.4%);

¹ H NMR (CDCl ₃ , 300 MHz): δ 7.75 (d, 1H), 7.57 (d, 1H), 6.60 (s, 1H), 6.45 (s, 1H), 4, 20 (m, 1H), 3.99 (s, 3H), 3.97 (s, 3H), 3.65 (dd, 1H), 3.36 (d, 1H), 3.20 (m, 1H ), 2.65 (m, 2H), 2.38 (s, 3H), 2.10 (m, 2H);

MS (ES +): m / z 482 (M + 1).

Example 19:

(+) - trans-2- (2,4-Dichloro-5-fluorophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one

Molten pyridine hydrochloride (5.5 g, 47.6 mmol) was added to the compound of formula (18) (0.53 g, 1.1 mmol) and heated at 180 ° C. for 1.5 hours. The reaction mixture was cooled to 25 ° C., diluted with MeOH (10 ml) and made basic using Na ₂ CO ₃ to pH 10. The mixture was filtered and the organic layer was concentrated. The residue was suspended in water (5 ml), stirred for 30 minutes, filtered and dried to give compound (+) - trans-2- (2,4-dichloro-5-fluorophenyl) -5,7-dihydroxy-8- ( 2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one.

Yield: 0.29 g (58%);

IR (KBr): 3422, 1664, 1618, 1401 cm ^-1 ;

¹ H NMR (CDCl ₃ + DMSO d ₆ , 300 MHz): δ 7.50 (m, 2H), 6.42 (s, 1H), 6.19 (s, 1H), 4.04 (m, 1H ), 3.71 (m, 2H), 3.16 (m, 2H), 2.88 (m, 1H), 2.58 (s, 3H), 2.36 (m, 1H), 1.92 (m, 1H);

MS (ES +): m / z 454 (M + 1).

Example 20:

(+) - trans-2- (2,4-Dichloro-5-fluorophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one hydrochloride

The compound of formula (19) (0.89 g, 1.96 mmol) was converted to its hydrochloride salt by suspension in IPA (5 ml) and 3.5% HCl (25 ml). The suspension was heated until a clear solution was obtained. The solution was cooled and the solid was filtered to obtain the compound (+) - trans-2- (2,4-dichloro-5-fluorophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chrome-4-one hydrochloride.

Yield: 0.91 g (92%);

¹ H NMR (CD ₃ OD, 300 MHz): δ 7.86 (m, 2H), 6.65 (s, 1H), 6.38 (s, 1H), 4.26 (q, 1H), 3 87 (m, 2H), 3.71 (m, 1H), 3.60 (dd, 1H), 3.56 (q, 1H), 2.99 (s, 3H), 2.55 (m, 1H); 2.28 (m, 1H); IR (KBr): 3386, 1657 cm ^-1 .

MS (ES +): m / z 454 (M-36.5);

Study: C ₂₁ H ₁₉ Cl ₃ FNO ₅ , C, 51.08 (51.35); H, 4.26 (3.87); N, 3.13 (2.85), Cl, 21.99 (21.70).

Example 21:

(+) - trans-2- (2,4-Dichloro-5-fluorophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one methanesulfonate

Methanesulfonic acid (0.012 g, 0.125 mmol) was added to a suspension of the compound of formula (19) (0.05 g, 0.11 mmol) in methanol (2 ml). It was stirred for 5 minutes at 25 ° C until a clear solution was obtained. The solvent was removed under reduced pressure to obtain a residue. The residue was washed twice with dry ether (2 × 5 ml) and dried under high vacuum (0.1 mm) to obtain compound (+) - trans-2- (2,4-dichloro-5-fluorophenyl) -5,7-dihydroxy -8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one methanesulfonate.

Yield: 0.054 g (90%);

Study: C ₂₂ H ₂₂ Cl ₂ FNO ₈ S · 2.5 H ₂ O C, 44.26 (44.34); H, 4.42 (4.53); N, 2.58 (2.35); Cl, 12.11 (11.92).

Example 22:

(+) - trans-2- (2,4-Dichloro-5-fluorophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one citrate

Citric acid monohydrate (0.023 g, 0.11 mmol) was added to a suspension of the compound of formula (19) (0.05 g, 0.11 mmol) in methanol (2 ml). It was stirred for 10 minutes at 50-55 ° C. The reaction mixture was concentrated and the solid was dried to give compound (+) - trans-2- (2,4-dichloro-5-fluorophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl ) -chrome-4-one citrate.

Yield: 0.065 g (91.5%);

Study: C ₂₇ H ₂₆ Cl ₂ FNO ₁₂ C, 49.75 (50.17); H, 4.25 (4.05); N, 2.60 (2.17); Cl, 10.69 (10.97).

Example 23:

(+) - trans-2- (2,4-Dichloro-5-fluorophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one tartrate

Tartaric acid (0.016 g, 0.11 mmol) was added to a suspension of the compound of formula (19) (0.05 g, 0.11 mmol) in methanol (2 ml). It was stirred for 10 minutes at 50-55 ° C. The reaction mixture was concentrated and the solid was dried to give compound (+) - trans-2- (2,4-dichloro-5-fluorophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl ) -chrome-4-one of tartrate.

Yield: 0.06 g (90.19%);

Study: C ₂₅ H ₂₄ Cl ₂ FNO ₁₁ · 2.0 H ₂ O C, 47.43 (46.84); H, 4.13 (4.37); N, 2.40 (2.18); Cl, 10.77 (11.08).

Example 24:

(+) - trans-2- (2,4-Dichloro-5-fluorophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one maleate

Maleic acid (0.0127 g, 0.11 mmol) was added to a suspension of the compound of formula (19) (0.05 g, 0.11 mmol) in methanol (2 ml). It was stirred for 5 minutes at 50 ° C. until a clear solution was obtained. The reaction mixture was concentrated and the solid was dried to give compound (+) - trans-2- (2,4-dichloro-5-fluorophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl ) -chrome-4-one maleate.

Yield: 0.059 g (90.19%);

Study: C ₂₅ H ₂₂ Cl ₂ FNO ₉ · 1.5 H ₂ O C, 50.54 (50.23); H, 4.08 (4.18); N, 2.31 (2.34); Cl, 11.83 (11.88).

Example 25:

(+) - trans-2- (2,4-Dichloro-5-fluorophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one acetate

Acetic acid (0.0065 ml, 0.0068 g, 0.11 mmol) was added to a suspension of the compound of formula (19) (0.05 g, 0.11 mmol) in methanol (2 ml). It was stirred for 5 minutes at 50-55 ° C until a clear solution was obtained. The reaction mixture was concentrated and the solid was dried to obtain the title compound.

Yield: 0.05 g (88.33%);

Study: C ₂₃ H ₂₂ Cl ₂ FNO ₉ C, 53.67 (53.71); H, 4.63 (4.31); N, 3.08 (2.72); Cl, 13.93 (13.79).

Example 26:

(+) - trans-2- (2,4-Dichloro-5-fluorophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one sulfate

A solution of H ₂ SO ₄ (5% methanol solution, 60 μl, 0.055 mmol) was added to a suspension of the compound of formula (19) (0.025 g, 0.055 mmol) in methanol (2 ml). It was stirred for 5 minutes at 50-55 ° C. The reaction mixture was concentrated and the solid was dried to obtain compound (+) - trans-2- (2,4-dichloro-5-fluoro phenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3- il) -chromen-4-one sulfate.

Yield: 0.025 g (82.24%);

MS (ES -): m / z 550 (M-1).

Example 27:

(+) - trans-2- (2,4-Dichloro-5-fluorophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one nitrate

A solution of HNO ₃ (5% methanol solution, 70 μl, 0.055 mmol) was added to a suspension of the compound of formula (19) (0.025 g, 0.055 mmol) in methanol (2 ml). It was stirred for 5 minutes at 50-55 ° C until a clear solution was obtained. The reaction mixture was concentrated and the solid was dried to give compound (+) - trans-2- (2,4-dichloro-5-fluorophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl ) -chrome-4-one nitrate.

Yield: 0.025 g (87.81%);

MS (ES-): m / z 515 (M-1).

Example 28:

(+) - trans-2,4-Dichlorobenzoic acid 2- (2-acetoxymethyl-1-methylpyrrolidin-3-yl) -6-acetyl-3,5-dimethoxyphenyl ester

A mixture of a compound of formula (6) (3 g, 8.9 mmol), 2,4-dichlorobenzoic acid (1.79 g, 9.3 mmol), DCC (3.87 g, 18.7 mmol) and DMAP (1,148 g, 9.3 mmol) was dissolved in dichloromethane (50 ml) and stirred at room temperature for 12 hours. The reaction mixture was cooled to 0 ° C. The precipitated dicyclohexylurea was filtered off, the organic layer was concentrated, and the residue was purified by column chromatography with 1% methanol in chloroform and 0.01% ammonia as an eluent to obtain the compound (+) - trans-2,4-dichlorobenzoic acid 2- (2-acetoxymethyl-1- methylpyrrolidin-3-yl) -6-acetyl-3,5-dimethoxyphenyl ester, which was used in the subsequent reaction without characterization.

Yield: 4.48 g (100%)

Example 29:

(+) - trans-2- (2,4-dichlorophenyl) -8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -5,7-dimethoxychromen-4-one

To a solution of n-BuLi (1.6 M in hexane, 10.67 ml, 17 mmol) in THF (25 ml), which was maintained at 0 ° C. under a nitrogen atmosphere, hexamethyldisilazane (3.56 ml, 17 mmol) was added with dropwise and stirred for 15 minutes. To this was added dropwise a solution of the compound of example (28) (4.48 g, 8.5 mmol) in THF (25 ml), maintaining the temperature at 0 ° C. After addition, the reaction was warmed to room temperature and stirred for 2.5 hours. The reaction mixture was acidified with diluted HCl and made basic with 10% sodium bicarbonate to pH 8-9. The aqueous layer was extracted with chloroform (3 × 50 ml). The organic layer was washed with water (25 ml), brine (25 ml) and dried over anhydrous Na ₂ SO ₄ . The organic layer was concentrated under reduced pressure and dried in vacuo to give 3- {3- [3- (2,4-dichlorophenyl) -3-ochopropionyl] -2-hydroxy-4,6-dimethoxyphenyl} -1-methylpyrrolidine- 2-yl-methyl ester as a brown solid (4.6 g), this ester was dissolved in conc. HCl (50 ml) and stirred for 3 hours to perform cyclization. At the end of 3 hours, the reaction mixture was basified with solid NaHCO ₃ to pH 8-9. The aqueous layer was extracted with chloroform (50 × 3 ml) and washed with water (25 ml) and brine (25 ml). The organic layer was dried over anhydrous Na ₂ SO ₄ ; concentrated under reduced pressure and dried under vacuum. The residue was purified by column chromatography with 3% methanol in chloroform and 0.01% ammonia as an eluent to obtain the compound, (+) - trans-2- (2,4-dichlorophenyl) -8- (2-hydroxymethyl-1-methylpyrrolidin-3 -yl) -5,7-dimethoxychromen-4-one as a yellow solid.

Yield: 3.2 g (81%);

¹ H NMR (CDCl ₃ , 300 MHz): δ 7.74 (d, 1H), 7.55 (d, 1H), 7.42 (dd, 1H), 6.56 (s, 1H), 6, 47 (s, 1H), 4.2 (m, 1H), 4.02 (s, 3H), 3.99 (s, 3H), 3.68 (dd, 1H), 3.39 (dd, 1H ), 3.23 (m, 1H), 2.75 (m, 1H), 2.63 (m, 1H), 2.40 (s, 3H), 2.07 (m, 2H); MS (ES +): m / z 464 (M + 1).

Example 30:

(+) - trans-2- (2,4-dichlorophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one

A mixture of the compound of Example (29) (3.2 g, 7.35 mmol), pyridine hydrochloride (3.2 g, 6.91 mmol) and a catalytic amount of quinoline was heated at 180 ° C. for a period of 2.5 hours. The reaction mixture was diluted with methanol (50 ml) and basified with solid Na ₂ CO ₃ to pH 10. The reaction mixture was filtered and washed with methanol. The organic layer was concentrated and the residue was purified by column chromatography using 0.01% ammonia and 3% methanol in chloroform as an eluent to obtain the compound (+) - trans-2- (2,4-dichlorophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one as a yellow solid.

Yield: 0.98 g (30%);

¹ H NMR (CDCl ₃ , 300 MHz): δ 12.5 (s, 1H), 7.54 (m, 2H), 7.40 (m, 1H), 6.40 (s, 1H), 6, 28 (s, 1H), 4.08 (m, 1H), 3.89 (m, 2H), 3.28 (m, 2H), 2.84 (m, 1H), 2.65 (s, 3H ), 2.45 (m, 1H); 1.95 (m, 1H);

MS (ES +): m / z 436 (M + 1).

Example 31:

(+) - trans-2- (2,4-dichlorophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one hydrochloride

The compound of formula (30) (0.4 g, 0.9 mmol) was suspended in methanol (2 ml) and treated with ethereal HCl and the organic solvent was evaporated to obtain the compound (+) - trans-2- (2,4-dichlorophenyl) - 5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one hydrochloride.

Yield: 0.4 g (92%);

¹ H NMR (CD ₃ OD, 300 MHz): δ 7.80 (d, 1H), 7.30 (d, 1H), 7.54 (dd, 1H), 6.58 (s, 1H), 6 , 37 (s, 1H), 4.25 (m, 1H), 3.90 (m, 2H), 3.59 (m, 2H), 3.41 (m, 1H), 2.98 (s, 3H); 2.52 (m, 1H); 2.29 (m, 1H);

MS (ES-): m / z 470 (M-1).

Example 32:

(+) - tract-4-bromo-2-chlorobenzoic acid 2- (2-acetoxymethyl-1-methylpyrrolidin-3-yl) -6-acetyl-3,5-dimethoxyphenyl ester

A mixture of the compound of example (6) (1.95 g, 5.5 mmol), 4-bromo-2-chlorobenzoic acid (1.44 g, 6.1 mmol), DCC (2.29 g, 11 mmol) and DMAP (0.68 g, 5.5 mmol) was dissolved in dichloromethane (50 ml) and stirred at room temperature for 12 hours. The reaction mixture was cooled to 0 ° C, the precipitated dicyclohexylurea was filtered off, the organic layer was concentrated, and the residue was purified by column chromatography with 1% methanol in chloroform and 0.01% ammonia as an eluent to obtain compound (+) - trans-4-bromo-2-chlorobenzoic 2- (2-acetoxymethyl-1-methylpyrrolidin-3-yl) -6-acetyl-3,5-dimethoxyphenyl ester acids.

Yield: 2.8 g (88.7%);

MS (ES +): m / z 569 (M + 1).

Example 33:

(+) - trans-2- (4-Bromo-2-chlorophenyl) -8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -5,7-dimethoxychromen-4-one

To a solution of n-BuLi (15% solution in hexane, 4.23 ml, 10 mmol) in THF (25 ml), which was maintained at 0 ° C. under nitrogen, hexamethyldisilazane (2.1 ml, 10 mmol) was added dropwise and stirred for 15 minutes. To this was added dropwise a solution of the compound of Example (32) (2.8 g, 5 mmol) in THF (25 ml), keeping the temperature at 0 ° C. After addition, the reaction was warmed to room temperature and stirred for 2.5 hours. The reaction mixture was acidified with dilute HCl and made basic with 10% sodium bicarbonate to pH 8-9. The aqueous layer was extracted with chloroform (3 × 50 ml). The organic layer was washed with water (25 ml), brine (25 ml) and dried over anhydrous Na ₂ SO ₄ . The organic layer was concentrated under reduced pressure and dried under vacuum to obtain 3- {3- [3- (4-bromo-2-chlorophenyl) -3-oxopropionyl] -2-hydroxy-4,6-dimethoxyphenyl} -1- methylpyrrolidin-2-yl-methyl ester as a brown solid (2.65 g). This ester was dissolved in conc. HCl (20 ml) and stirred for 3 hours to complete cyclization. At the end of 3 hours, the reaction mixture was basified with solid NaHCO ₃ to pH 8-9. The aqueous layer was extracted with chloroform (50 × 3 ml) and washed with water (25 ml) and brine (25 ml). The organic layer was dried over anhydrous Na ₂ SO ₄ , concentrated under reduced pressure, and dried under vacuum. The residue was purified by column chromatography with 3% methanol in chloroform and 0.01% ammonia as an eluent to obtain the compound, (+) - trans-2- (4-bromo-2-chlorophenyl) -8- (2-hydroxymethyl-1-methylpyrrolidine -3-yl) -5,7-dimethoxychromen-4-one as a yellow solid.

Yield: 1.4 g (59%);

¹ H NMR (CDCl ₃ , 300 MHz): δ 7.69 (d, 1H), 7.66 (d, 1H), 7.55 (dd, 1H), 6.55 (s, 1H), 6, 45 (s, 1H), 4.19 (m, 1H), 4.00 (s, 3H), 3.98 (s, 3H), 3.66 (dd, 1H), 3.38 (m, 1H ), 3.23 (m, 1H), 2.75 (m, 1H), 2.63 (m, 1H), 2.40 (s, 3H), 2.10 (m, 2H);

MS (ES +): m / z 509 (M + 1).

Example 34:

(+) - trans-2- (4-Bromo-2-chlorophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one

A mixture of the compound of Example (33) (1.4 g, 2.75 mmol), pyridine hydrochloride (1.4 g, 12 mmol) and a catalytic amount of quinoline was heated at 180 ° C. for a period of 2.5 hours. The reaction mixture was diluted with methanol (50 ml) and basified with solid Na ₂ CO ₃ to pH 10. The reaction mixture was filtered and washed with methanol. The organic layer was concentrated and the residue was purified by column chromatography using 0.01% ammonia and 3% methanol in chloroform as an eluent to obtain the compound (+) - trans-2- (4-bromo-2-chlorophenyl) -5,7-dihydroxy- 8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one as a yellow solid.

Yield: 0.88 g (60.6%);

¹ H NMR (CDCl ₃ ; 300 MHz): δ 7.91 (m, 1H), 7.87 (m, 1H), 7.69 (m, 1H), 6.41 (s, 1H), 6, 15 (s, 1H), 4.03 (m, 1H), 3.77 (dd, 1H), 3.66 (dd, 1H), 3.47 (m, 1H), 3.33 (m, 1H ), 3.18 (m, 1H), 2.76 (s, 3H), 2.24 (m, 2H);

MS (ES +): m / z 481 (M + 1).

Example 35:

(+) - trans-2- (4-bromo-2-chlorophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one hydrochloride

The compound of formula (34) (0.48 g, 1.0 mmol) was suspended in methanol (2 ml) and treated with ethereal HCl and the organic solvent was evaporated to give the compound (+) - trans-2- (4-bromo-2-chlorophenyl) ) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one hydrochloride.

Yield: 0.4 g (93%);

¹ H NMR (CD ₃ OD, 300 MHz): δ 7.87 (m, 1H), 7.71 (m, 2H), 6.58 (s, 1H), 6.38 (s, 1H), 4 28 (m, 1H), 3.90 (m, 2H), 3.73 (m, 1H), 3.59 (m, 1H), 3.45 (m, 1H), 2.99 (s, 3H); 2.58 (m, 1H); 2.28 (m, 1H);

MS (ES-): m / z 515 (M-1).

Example 36:

(+) - trans-2-Chloro-4-cyanobenzoic acid 2- (2-acetoxymethyl-1-methylpyrrolidin-3-yl) -6-acetyl-3,5-dimethoxyphenyl ester

A mixture of the compound of example (6) (3.84 g, 10 mmol), 2-chloro-4-cyanobenzoic acid (2.48 g, 13 mmol), DCC (4.51 g, 21 mmol) and DMAP (1.34 g, 10 mmol) was dissolved in dichloromethane (50 ml) and stirred at room temperature for 12 hours. The reaction mixture was cooled to 0 ° C, the precipitated dicyclohexylurea was filtered off, the organic layer was concentrated, and the residue was purified by column chromatography with 1% methanol in chloroform and 0.01% ammonia as an eluent to obtain the compound (+) - trans-2-chloro-4-cyanobenzoic 2- (2-acetoxymethyl-1-methylpyrrolidin-3-yl) -6-acetyl-3,5-dimethoxyphenyl ester acids.

Yield: 3.5 g (62%);

¹ H NMR (CDCl ₃ , 300 MHz): δ 8.19 (d, 1H), 7.79 (s, 1H), 7.69 (d, 1H), 6.45 (s, 1H), 4, 10 (m, 1H), 3.93 (s, 6H), 3.48 (m, 2H), 3.11 (m, 1H), 2.72 (m, 1H), 2.48 (s, 3H ), 2.35 (s, 3H), 2.10 (m, 1H), 1.92 (m, 2H), 1.74 (s, 3H);

MS (ES +): m / z 515 (M + 1).

Example 37:

(+) - trans-2- (2-Chloro-4-cyanophenyl) -8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -5,7-dimethoxychromen-4-one

To a solution of n-BuLi (1.6 M solution in hexane, 17 ml, 27 mmol) in THF (50 ml), which was maintained at 0 ° C. under nitrogen, hexamethyldisilazane (5.7 ml, 27 mmol) was added dropwise and mixed for 15 minutes. To this was added dropwise a solution of the compound of example (36) (5.6 g, 10 mmol) in THF (50 ml), maintaining the temperature at 0 ° C. After addition, the reaction was warmed to room temperature and stirred for 2.5 hours. The reaction mixture was acidified with dilute HCl and made basic with 10% sodium bicarbonate to pH 8-9. The aqueous layer was extracted with chloroform (3 × 75 ml). The organic layer was washed with water (50 ml), brine (50 ml) and dried over anhydrous Na ₂ SO ₄ . The organic layer was concentrated under reduced pressure and dried under vacuum to obtain 3- {3- [3- (2-chloro-4-cyanophenyl) -3-oxopropionyl] -2-hydroxy-4,6-dimethoxyphenyl} -1- methylpyrrolidin-2-yl-methyl ester as an oil (6.1 g). This ester was dissolved in conc. HCl (60 ml) and stirred for 3 hours to complete cyclization. At the end of 3 hours, the reaction mixture was basified with solid NaHCO ₃ to pH 8-9. The aqueous layer was extracted with chloroform (50 × 3 ml) and washed with water (25 ml) and brine (25 ml). The organic layer was dried over anhydrous Na ₂ SO ₄ , concentrated under reduced pressure, and dried under vacuum. The residue was purified by column chromatography with 3% methanol in chloroform and 0.01% ammonia as an eluent to obtain the compound (+) - trans-2- (2-chloro-4-cyanophenyl) -8- (2-hydroxymethyl-1-methylpyrrolidine- 3-yl) -5,7-dimethoxychromen-4-one as a yellow solid.

Yield: 0.75 g (15%);

IR (KBr): 3431, 2233, 1648, 1599 cm ^-1 ;

¹ H NMR (CDCl ₃ , 300 MHz): δ 8.20 (d, 1H), 8.04 (s, 1H), 7.88 (d, 1H), 6.65 (s, 1H), 6, 46 (s, 1H), 4.25 (m, 1H), 4.01 (s, 3H), 3.99 (s, 3H), 3.70 (m, 1H), 3.35 (m, 2H ), 2.75 (m, 2H), 2.43 (s, 3H), 2.10 (m, 2H);

MS (ES +): m / z 455 (M + 1).

Example 38:

(+) - trans-2- (2-Chloro-4-cyanophenyl) -8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -5,7-dimethoxychromen-4-one hydrochloride

The compound of formula (37) (20 mg, 0.046 mmol) was suspended in methanol (2 ml) and treated with ethereal HCl and the organic solvent was evaporated to give compound (+) - trans-2- (2-chloro-4-cyanophenyl) -8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -5,7-dimethoxychromen-4-one hydrochloride.

Yield: 20 mg (98%);

¹ H NMR (CD ₃ OD, 300 MHz): δ 8.08 (m, 1H), 7.97 (m, 1H), 7.86 (m, 1H), 6.78 (s, 1H), 6 58 (s, 1H), 4.32 (m, 1H), 4.10 (s, 3H), 4.01 (s, 3H), 3.84 (m, 2H), 3.75 (m, 1H), 3.57 (m, 1H), 3.41 (m, 1H), 2.98 (s, 3H), 2.43 (m, 1H), 2.31 (m, 1H); MS (ES-): m / z 489 (M-1)

Example 39:

(+) - trans-2- (2-Chloro-4-cyanophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one

A mixture of the compound of Example (37) (0.25 g, 0.55 mmol), pyridine hydrochloride (0.25 g, 2.2 mmol) and a catalytic amount of quinoline was heated at 180 ° C. for a period of 2.5 hours. The reaction mixture was diluted with methanol (50 ml) and basified with solid Na ₂ CO ₃ to pH 10. The reaction mixture was filtered and washed with methanol. The organic layer was concentrated and the residue was purified by column chromatography using 0.01% ammonia and 4.5% methanol in chloroform as an eluent to obtain compound (+) - trans-2- (2-chloro-4-cyanophenyl) -5,7- dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one as a yellow solid.

Yield: 0.11 g (49.5%);

IR (KBr): 3396, 2234, 1656, 1615 cm ^-1 ;

¹ H NMR (CD ₃ OD, 300 MHz): δ 8.05 (s, 1H), 7.90 (m, 2H), 6.45 (s, 1H), 6.11 (s, 1H), 4 02 (m, 1H), 3.75 (dd, 2H), 3.49 (m, 2H), 3.21 (m, 1H), 2.76 (s, 3H), 2.23 (m, 2H);

MS (ES +): m / z 427 (M + 1)

Example 40:

(+) - trans-2- (2-Chloro-4-cyanophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one hydrochloride

The compound of formula (39) (0.1 g, 0.23 mmol) was suspended in methanol (2 ml) and treated with ethereal HCl and the organic solvent was evaporated to obtain the compound (+) - trans-2- (2-chloro-4-cyanophenyl ) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one hydrochloride.

Yield: 0.1 g (92.6%);

MS (ES-): m / z 461 (M-1).

Example 41:

(+) - trans-2-chloro-4-trifluoromethylbenzoic acid 2- (2-acetoxymethyl-1-methylpyrrolidin-3-yl) -6-acetyl-3,5-dimethoxyphenyl ester

A mixture of the compound of example (6) (1.16 g, 3.2 mmol), 2-chloro-4-trifluoromethylbenzoic acid (0.88 g, 4 mmol), DCC (1.35 g, 6.5 mmol) and DMAP (0.4 g, 3.27 mmol) was dissolved in dichloromethane (50 ml) and stirred at room temperature for 12 hours. The reaction mixture was cooled to 0 ° C, the precipitated dicyclohexylurea was filtered off, the organic layer was concentrated, and the residue was purified by column chromatography with 1% methanol in chloroform and 0.01% ammonia as an eluent to obtain the compound (+) - trans-2-chloro-4-trifluoromethylbenzene 2- (2-acetoxymethyl-1-methylpyrrolidin-3-yl) -6-acetyl-3,5-dimethoxyphenyl ester acids.

Yield: 1.44 g (78.8%);

¹ H NMR (CDCl ₃ , 300 MHz): δ 8.19 (d, 1H), 7.79 (s, 1H), 7.69 (d, 1H), 6.45 (s, 1H), 4, 14 (m, 1H), 3.93 (s, 6H), 3.52 (m, 1H), 3.17 (m, 1H), 2.63 (m, 1H), 2.48 (s, 3H ), 2.41 (s, 3H), 2.15 (m, 1H), 2.05 (m, 1H), 1.81 (s, 3H), 1.62 (m, 2H);

MS (ES +): m / z 558 (M + 1)

Example 42:

(+) - trans-2- (2-Chloro-4-trifluoromethylphenyl) -8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -5,7-dimethoxychromen-4-one

To a solution of n-BuLi (15% solution in hexane, 2.2 ml, 5 mmol) in THF (10 ml), which was maintained at 0 ° C. under a nitrogen atmosphere, hexamethyldisilazane (1.08 ml, 5.1 mmol) was added dropwise and stirred for 15 minutes To this was added dropwise a solution of the compound of example (41) (1.44 g, 2.5 mmol) in THF (10 ml), maintaining the temperature at 0 ° C. After addition, the reaction was warmed to room temperature and stirred for 2.5 hours. The reaction mixture was acidified with dilute HCl and made basic with 10% sodium bicarbonate to pH 8-9. The aqueous layer was extracted with chloroform (3 × 25 ml). The organic layer was washed with water (25 ml), brine (25 ml) and dried over anhydrous Na ₂ SO ₄ . The organic layer was concentrated under reduced pressure and dried in vacuo to give acetic acid 3- {3- [3- (2-chloro-4-trifluoromethylphenyl) -3-oxopropionyl] -2-hydroxy-4,6-dimethoxyphenyl} -1- methylpyrrolidin-2-yl-methyl ester as an oil (1.3 g, 90.2%). This ester was dissolved in conc. HCl (10 ml) and stirred for 3 hours to perform cyclization. At the end of 3 hours, the reaction mixture was basified with solid NaHCO ₃ to pH 8-9. The aqueous layer was extracted with chloroform (25 × 3 ml) and washed with water (25 ml) and brine (25 ml). The organic layer was dried over anhydrous Na ₂ SO ₄ ; concentrated under reduced pressure and dried under vacuum. The residue was purified by column chromatography with 3% methanol in chloroform and 0.1% ammonia as an eluent to obtain the compound (+) - trans-2- (2-chloro-4-trifluoromethylphenyl) -8- (2-hydroxymethyl-1-methylpyrrolidine- 3-yl) -5,7-dimethoxychromen-4-one as a yellow solid.

Yield: 0.56 g (48.2%);

¹ H NMR (CDCl ₃ , 300 MHz): δ 7.95 (d, 1H), 7.78 (s, 1H), 7.69 (d, 1H), 6.61 (s, 1H), 6, 46 (s, 1H), 4.21 (m, 1H), 4.01 (s, 3H), 3.93 (s, 3H), 3.71 (dd, 1H), 3.41 (d, 1H ), 3.26 (m, 1H), 2.84 (m, 1H), 2.70 (m, 1H), 2.44 (s, 3H), 2.10 (m, 2H); MS (ES +): m / z 497 (M + 1)

Example 43:

(+) - trans-2- (2-Chloro-4-trifluoromethyl-phenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one

A mixture of the compound of Example (42) (0.25 g, 0.5 mmol), pyridine hydrochloride (0.25 g, 2.16 mmol) and a catalytic amount of quinoline was heated at 180 ° C. for a period of 2.5 hours. The reaction mixture was diluted with methanol (25 ml) and basified with solid Na ₂ CO ₃ to pH 10. The reaction mixture was filtered and washed with methanol. The organic layer was concentrated and the residue was purified by column chromatography using 0.1% ammonia and 4.5% methanol in chloroform as an eluent to obtain compound (+) - trans-2- (2-chloro-4-trifluoromethylphenyl) -5,7- dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one as a yellow solid.

Yield: 0.15 g (63.7%);

¹ H NMR (CDCl ₃ , 300 MHz): δ 7.99 (m, 2H), 7.83 (d, 1H), 6.65 (s, 1H), 6.41 (s, 1H), 4, 24 (m, 1H), 3.90 (m, 2H), 3.70 (m, 1H), 3.60 (m, 1H), 3.41 (m, 1H), 2.99 (s, 3H ), 2.54 (m, 1H); 2.28 (m, 1H);

MS (ES +): m / z 470 (M + 1)

Example 44:

(+) - trans-2- (2-Chloro-4-trifluoromethylphenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one hydrochloride

The compound of formula (43) (0.1 g, 0.2 mmol) was suspended in methanol (2 ml) and treated with ethereal HCl and the organic solvent was evaporated to give the compound (+) - trans-2- (2-chloro-4-trifluoromethyl) -phenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one hydrochloride.

Yield: 0.1 g (92.8%);

¹ H NMR (CDCl ₃ , 300 MHz): δ 8.02 (d, 2H), 7.83 (d, 1H), 6.64 (s, 1H), 6.41 (s, 1H), 4, 23 (m, 1H), 3.73 (m, 2H), 3.68 (m, 1H), 3.51 (m, 1H), 3.39 (m, 1H), 2.99 (s, 3H ), 2.54 (m, 1H), 2.31 (m, 1H).

Example 45:

(+) - trans-2-chloro-4-nitrobenzoic acid 2- (2-acetoxymethyl-1-methylpyrrolidin-3-yl) -6-acetyl-3,5-dimethoxyphenyl ester

2-Chloro-4-nitrobenzoic acid (13 g, 64 mmol) and thionyl chloride (40 ml) were heated together at 85 ° C for 3 hours. The excess thionyl chloride was evaporated under reduced pressure, and the residue was dried in vacuo and dissolved in THF (100 ml). This was added dropwise to a solution of the compound of Example (6) (15 g, 42 mmol) and triethylamine (29.5 ml, 213 mmol) in THF (100 ml), which was maintained at 15-20 ° C. The reaction mixture was warmed to room temperature and stirred for 12 hours. At the end of 12 hours, the reaction mixture was concentrated to remove THF, acidified with dilute HCl, basified with 10% NaHCO ₃ solution to pH 10, extracted with EtOAc (3 × 100 ml), washed with water (50 ml), brine (50 ml) and the organic layer dried over anhydrous Na ₂ SO ₄ . The organic layer was concentrated under reduced pressure to obtain the compound (+) - trans-2-chloro-4-nitrobenzoic acid 2- (2-acetoxymethyl-1-methylpyrrolidin-3-yl) -6-acetyl-3,5-dimethoxyphenyl ester as an oil, which was used by itself in the next reaction without purification.

Yield: 24.8 g (71.7%);

¹ H NMR (CDCl ₃ , 300 MHz): δ 8.35 (s, 1H), 8.25 (m, 2H), 6.47 (s, 1H), 4.17 (m, 1H), 3, 94 (s, 6H), 3.59 (m, 1H), 3.17 (m, 1H), 2.80 (m, 1H), 2.49 (s, 6H), 2.46 (m, 2H ), 2.17 (m, 1H), 1.95 (m, 1H), 1.83 (s, 3H);

MS (ES +): m / z 535 (M + 1).

Example 46:

(+) - trans-2- (2-Chloro-4-nitrophenyl) -8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -5,7-dimethoxychromen-4-one

A solution of the compound of example (45) (20 g, 37.5 mmol) in dioxane (150 ml) was added dropwise to a suspension of sodium hydride (60% dispersion, 5.68 g, 142 mmol) in dioxane (50 ml) and the reaction mixture stirred at room temperature for 12 hours. The reaction mixture was quenched by the addition of methanol dropwise and the solvents were evaporated under reduced pressure. The residue was acidified with dilute HCl, basified with 10% NaHCO ₃ solution to pH 8-9 and extracted with EtOAc (3 × 250 ml). The organic layer was washed with water (50 ml), brine (50 ml), dried over anhydrous Na ₂ SO ₄ and concentrated under reduced pressure to obtain 3- {3- [3- (2-chloro-4-nitrophenyl) -3 acetic acid -oxopropionyl] -2-hydroxy-4,6-dimethoxyphenyl} -1-methylpyrrolidin-2-yl-methyl ester as an oil (18.2 g). This ester was dissolved in conc. HCl (50 ml) and stirred at room temperature for 3 hours. At the end of 3 hours, the reaction mixture was basified with solid NaHCO ₃ to pH 8-9. The aqueous layer was extracted with chloroform (3 × 250 ml) and washed with water (50 ml) and brine (50 ml). The organic layer was dried over anhydrous Na ₂ SO ₄ ; concentrated under reduced pressure and dried under vacuum. The residue was purified by column chromatography with 3% methanol in chloroform and 0.1%) ammonia as an eluent to obtain the compound (+) - trans-2- (2-chloro-4-nitrophenyl) -8- (2-hydroxymethyl-1-methylpyrrolidine -3-yl) -5,7-dimethoxychromen-4-one as a pale yellow solid.

Yield: 9.5 g (53.5%);

IR (KBr): 3447, 1648, 1600, 1570 cm ^-1 ;

¹ H NMR (CDCl ₃ , 300 MHz): δ 8.37 (d, 1H), 8.26 (dd, 1H), 8.09 (d, 1H), 6.67 (s, 1H), 6, 46 (s, 1H), 4.18 (m, 1H), 4.01 (s, 3H), 3.98 (s, 3H), 3.67 (m, 1H), 3.32 (m, 1H ), 3.19 (m, 1H), 2.62 (m, 2H), 2.36 (s, 3H), 2.04 (m, 2H);

MS (ES +): m / z 475 (M + 1)

Example 47:

(+) - trans-2- (2-Chloro-4-nitrophenyl) -8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -5,7-dimethoxychromen-4-one hydrochloride

The compound of formula (46) (0.05 g, 0.105 mmol) was suspended in methanol (2 ml) and treated with ethereal HCl and the organic solvent was evaporated to obtain the compound (+) - trans-2- (2-chloro-4-nitrophenyl) - 8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -5,7-dimethoxychromen-4-one hydrochloride.

Yield: 0.05 g (53.7%); ¹ H NMR (CD ₃ OD, 300 MHz): δ 8.50 (d, 1H), 8.27 (dd, 1H), 8.09 (d, 1H), 6.79 (s, 1H), 6 62 (s, 1H), 4.28 (m, 1H), 4.11 (s, 3H), 4.02 (s, 3H), 3.88 (m, 2H), 3.72 (m, 1H), 3.50 (m, 2H), 2.99 (s, 3H), 2.49 (m, 1H), 2.26 (m, 1H); MS (ES +): m / z 511 (M + 1).

Example 48:

(+) - trans-2- (2-Chloro-4-nitrophenyl) -8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -5,7-dimethoxychromen-4-one gluconate

The compound of formula (46) (6.4 g, 13.4 mmol) was suspended in methanol (300 ml) and treated with gluconic acid (2.63 g, 13.4 mmol) and the organic solvent was evaporated to obtain the compound (+) - trans-2 - (2-chloro-4-nitro phenyl) -8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -5,7-dimethoxychromen-4-one gluconate.

Yield: 8.5 g (93.9%);

m.p .: 137-140 ° C;

IR (KBr): 3404, 1648, 1601, 1524 cm ^-1 ;

¹ H NMR (CD ₃ OD, 300 MHz): δ 8.50 (d, 1H), 8.33 (dd, 1H), 8.09 (d, 1H), 6.78 (s, 1H), 6 61 (s, 1H), 4.31 (m, 1H), 4.15 (s, 3H), 4.01 (s, 3H), 3.72 (m, 11H), 2.95 (s, 3H); 2.4 (m, 1H); 2.30 (m, 1H); [α] _D ²⁵ : + 11.76 ° (c = 0.76, MeOH);

MS (ES +): m / z 475 (M + 1), free base.

Example 49:

(+) - trans-2- (2-Chloro-4-nitrophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one

A mixture of the compound of Example (46) (4.2 g, 8.8 mmol), pyridine hydrochloride (4.2 g, 36 mmol) and a catalytic amount of quinoline was heated at 180 ° C. for a period of 2 hours. The reaction mixture was cooled and diluted with methanol (50 ml) and made basic with solid Na ₂ CO ₃ to pH 10. The reaction mixture was filtered and washed with methanol. The organic layer was concentrated and the residue was purified by column chromatography using 0.1% ammonia and 3% methanol in chloroform as an eluent to obtain the compound (+) - trans-2- (2-chloro-4-nitrophenyl) -5,7-dihydroxy- 8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one as a yellow solid.

Yield: 2.8 g (70.88%);

¹ H NMR (CDCl ₃ , 300 MHz): δ 8.48 (d, 1H), 8.34 (dd, 1H), 8.02 (d, 1H), 6.48 (s, 1H), 6, 12 (s, 1H), 4.03 (m, 1H), 3.74 (m, 2H), 3.64 (m, 1H), 3.60 (m, 1H), 3.22 (m, 1H ), 2.77 (s, 3H); 2.22 (m, 2H);

MS (ES +): m / z 447 (M + 1)

Example 50:

(+) - trans-2- (2-Chloro-4-nitrophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one hydrochloride

The compound of formula (49) (2.8 g, 6.2 mmol) was suspended in methanol (10 ml) and treated with ethereal HCl and the organic solvent was evaporated to obtain the compound (+) - trans-2- (2-chloro-4-nitrophenyl) - 5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one hydrochloride.

Yield: 2.7 g (92.5%);

¹ H NMR (CD ₃ OD, 300 MHz): δ 8.51 (d, 1H), 8.36 (dd, 1H), 8.09 (d, 1H), 6.69 (s, 1H), 6 41 (s, 1H), 4.25 (m, 1H), 3.91 (m, 2H), 3.73 (m, 1H), 3.70 (m, 1H), 3.63 (m, 1H), 3.00 (s, 3H), 2.55 (m, 1H), 2.32 (m, 1H);

MS (ES +): m / z 483 (M + 1);

[α] _D ²⁵ : + 13.0 ° (c = 0.2, methanol.

Example 51:

(+) - trans-2- (4-amino-2-chlorophenyl) -8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -5,7-dimethoxychromen-4-one

The compound of formula (46) (1.0 g, 2.1 mmol), tin chloride (1.9 g, 8.4 mmol) and EtOAc (5 ml) were stirred together at room temperature for 12 hours. At the end of 12 hours, the reaction mixture was basified with 10% NaOH to pH 10 and extracted with EtOAc (3 × 10 ml). The organic layers were combined, washed with water (10 ml), brine (10 ml), dried over anhydrous Na ₂ SO ₄ and concentrated under reduced pressure to obtain the compound (+) - trans-2- (4-amino-2-chlorophenyl) - 8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -5,7-dimethoxychromen-4-one as a solid.

Yield: 0.74 g (79%);

MS (ES +): 445 (M + 1).

Example 52:

(+) - trans-2- (4-amino-2-chlorophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one (compound-1)

(+) - trans-2- (2-Chloro-4-methylaminophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one (compound-2)

A mixture of the compound of Example (51) (0.7 g, 1.57 mmol), pyridine hydrochloride (0.7 g, 6 mmol) and a catalytic amount of quinoline was heated at 180 ° C. for a period of 2.5 hours. The reaction mixture was cooled and diluted with methanol (25 ml) and basified with solid Na ₂ CO ₃ to pH 10. The reaction mixture was filtered and washed with methanol. The organic layer was concentrated and the residue was purified by column chromatography using 0.1% ammonia and 3% methanol in chloroform as an eluent to obtain compounds (+) - trans-2- (4-amino-2-chlorophenyl) -5,7-dihydroxy- 8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one (compound-1) and (+) - trans-2- (2-chloro-4-methylaminophenyl) -5,7-dihydroxy -8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one (compound-2) as yellow solids.

Compound-1: Yield: 0.25 g (38%); MS (ES +): 417 (M + 1).

Compound-2: Yield: 0.13 g (19%); MS (ES +): 431 (M + 1).

Example 53:

(+) - trans-2- (4-amino-2-chlorophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one hydrochloride

Compound (+) - trans-2- (4-amino-2-chlorophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one (0.25 g, 0.6 mmol) was suspended in methanol (2 ml) and treated with ethereal HCl and the organic solvent was evaporated to obtain the compound (+) - trans-2- (4-amino-2-chloro phenyl) -5,7-dihydroxy- 8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one hydrochloride.

Yield: 0.26 g (96%);

¹ H NMR (CD ₃ OD, 300 MHz): δ 7.78 (d, 1H), 7.26 (d, 1H), 7.13 (dd, 1H), 6.56 (s, 1H), 6 36 (s, 1H), 4.30 (m, 1H), 3.91 (m, 2H), 3.80 (m, 1H), 3.60 (dd, 1H), 3.46 (m, 1H), 3.00 (s, 3H), 2.52 (m, 1H), 2.29 (m, 1H);

MS (ES +): 417 (M + 1), free base; MS (ES-): 451 (M-1)

Example 54:

(+) - trans-2- (2-Chloro-4-methylaminophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one hydrochloride

Compound (+) - trans-2- (2-Chloro-4-methylaminophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one (0.13 g, 0.3 mmol) was suspended in methanol (2 ml) and treated with ethereal HCl and the organic solvent was evaporated to give the salt (+) - trans-2- (2-chloro-4-methylaminophenyl) -5,7-dihydroxy-8 - (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one hydrochloride.

Yield: 0.13 g (96%);

¹ H NMR (CD ₃ OD, 300 MHz): δ 7.67 (d, 1H), 6.86 (d, 1H), 6.80 (dd, 1H), 6.54 (s, 1H), 6 34 (s, 1H), 4.32 (m, 1H), 3.94 (m, 2H), 3.64 (m, 1H), 3.58 (dd, 1H), 3.44 (m, 1H), 3.00 (s, 3H), 2.86 (s, 3H), 2.53 (m, 1H), 2.33 (m, 1H);

MS (ES +): 431 (M + 1), free base; MS (ES-): 465 (M-1).

Example 55:

(+) - trans-2-chloro-4-pyrrolidin-1-yl-benzoic acid 2- (2-acetoxymethyl-1-methylpyrrolidin-3-yl) -6-acetyl-3,5-dimethoxyphenyl ester

2-Chloro-4-pyrrolidin-1-yl-benzoic acid (1.0 g, 4.4 mmol), thionyl chloride (0.35 ml, 4.8 mmol), N-methylpyrrolidin-2-one (0.1 ml) and dichloromethane (20 ml) was stirred together at room temperature for 12 hours. To this solution was added a compound of formula (6) (0.86 g, 2.4 mmol) and the mixture was cooled to 0-5 ° C. After cooling, triethylamine (1.7 ml, 12.7 mmol) was added and the reaction mixture was warmed to room temperature and stirred for 4 hours. The mixture was diluted with chloroform (25 ml), washed with 10% NaHCO ₃ (10 ml), water (10 ml), brine (10 ml) and the organic layer was dried over anhydrous Na ₂ SO ₄ . The organic layer was concentrated under reduced pressure to obtain the compound (+) - trans-2-chloro-4-pyrrolidin-1-ylbenzoic acid 2- (2-acetoxymethyl-1-methylpyrrolidin-3-yl) -6-acetyl-3,5 β-dimethoxyphenyl ester as a solid, which was used as such in the next reaction without purification.

Yield: 1.37 g (100%);

MS (ES +): 559 (M + 1).

Example 56:

(+) - trans-2- (2-Chloro-4-pyrrolidin-1-yl-phenyl) -8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -5,7-dimethoxychromen-4-one

To a solution of n-BuLi (1.6 M in hexane, 3 ml, 4.8 mmol) in THF (10 ml), which was maintained at 0 ° C. under a nitrogen atmosphere, hexamethyldisilazane (1 ml, 4.75 mmol) was added with dropwise and stirred for 15 minutes. To this was added dropwise a solution of the compound of Example (55) (1.37 g, 2.45 mmol) in THF (5 ml), maintaining the temperature at 0 ° C. After addition, the reaction mixture was warmed to room temperature and stirred for 2.5 hours. The reaction mixture was acidified with dilute HCl and made basic with 10% sodium bicarbonate to pH 8-9. The aqueous layer was extracted with chloroform (3 × 50 ml). The organic layer was washed with water (25 ml), brine (25 ml) and dried over anhydrous Na ₂ SO ₄ . The organic layer was concentrated under reduced pressure and dried in vacuo to give acetic acid 3- {3- [3- (2-chloro-4-pyrrolidin-1-yl-phenyl) -3-oxopropionyl] -2-hydroxy-4,6 -dimethoxyphenyl} -1-methylpyrrolidin-2-yl-methyl ester as a brown solid (0.97 g, 70.8%). This ester was dissolved in 5 ml conc. HCl and stirred for 3 hours to perform cyclization. At the end of 3 hours, the reaction mixture was basified with solid Na ₂ CO ₃ to pH 10. The aqueous layer was extracted with chloroform (3 × 25 ml) and washed with water (10 ml) and brine (10 ml). The organic layer was dried over anhydrous Na ₂ SO ₄ ; concentrated under reduced pressure and dried under vacuum. The residue was purified by column chromatography with 3% methanol in chloroform and 0.1% ammonia as an eluent to obtain the compound (+) - trans-2- (2-chloro-4-pyrrolidin-1-yl-phenyl) -8- (2- hydroxymethyl-1-methylpyrrolidin-3-yl) -5,7-dimethoxychromen-4-one as a brown solid.

Yield: 0.4 g (46.5%);

¹ H NMR (CDCl ₃ , 300 MHz): δ 7.62 (d, 1H), 6.58 (m, 2H), 6.50 (m, 1H), 6.42 (s, 1H), 4, 24 (m, 1H), 3.99 (s, 3H), 3.96 (s, 3H), 3.68 (m, 1H), 3.48 (m, 1H), 3.32 (m, 5H ), 2.88 (m, 1H), 2.77 (m, 1H), 2.53 (s, 3H), 2.05 (m, 6H);

MS (ES +): 499 (M + 1).

Example 57:

(+) - trans-2- (2-Chloro-4-pyrrolidin-1-yl-phenyl) -5,7-dihydroxy-8- (2-hydroxy methyl-1-methylpyrrolidin-3-yl) -chrome-4 -it

A mixture of the compound of Example (56) (0.4 g, 0.8 mmol), pyridine hydrochloride (0.4 g, 3.46 mmol) and a catalytic amount of quinoline was heated at 180 ° C. for a period of 2.5 hours. The reaction mixture was diluted with methanol (50 ml) and basified with solid Na ₂ CO ₃ to pH 10. The reaction mixture was filtered and washed with methanol. The organic layer was concentrated and the residue was purified by column chromatography using 0.1% ammonia and 3% methanol in chloroform as an eluent to obtain the compound (+) - trans-2- (2-chloro-4-pyrrolidin-1-yl-phenyl) - 5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one as a yellow solid.

Yield: 23 mg (6.6%);

¹ H NMR (CDCl ₃ , 300 MHz): δ 12.82 (s, 1H), 7.47 (d, 1H), 6.62 (d, 1H), 6.52 (dd, 1H), 6, 45 (s, 1H), 6.30 (s, 1H), 4.2 (m, 1H), 3.88 (m, 2H), 3.34 (m, 5H), 2.96 (m, 1H ), 2.68 (s, 3H), 2.40 (m, 1H), 2.09 (m, 6H);

MS (ES +): 471 (M + 1).

Example 58:

(+) - trans-2- (2-Chloro-4-pyrrolidin-1-yl-phenyl) -5,7-dihydroxy-8- (2-hydroxy methyl-1-methylpyrrolidin-3-yl) -chrome-4 -one hydrochloride

The compound of formula (57) (23 mg, 0.049 mmol) was suspended in methanol (2 ml) and treated with ethereal HCl and the organic solvent was evaporated to give the salt (+) - trans-2- (2-chloro-4-pyrrolidin-1-yl phenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one hydrochloride.

Yield: 23 mg (92.8%);

¹ H NMR (CD ₃ OD, 300 MHz): δ 7.66 (d, 1H), 6.69 (d, 1H), 6.62 (dd, 1H), 6.54 (s, 1H), 6 36 (s, 1H), 4.32 (m, 1H), 3.90 (m, 2H), 3.71 (m, 1H), 3.57 (m, 1H), 3.61 (m, 1H), 3.37 (m, 4H), 3.06 (s, 3H), 2.54 (m, 1H), 2.29 (m, 1H)), 2.06 (m, 4H); MS (ES-): 505 (M-1).

Example 59:

(+) - trans-2-chloro-4-isopropylaminobenzoic acid 2- (2-acetoxymethyl-1-methylpyrrolidin-3-yl) -6-acetyl-3,5-dimethoxyphenyl ester

2-Chloro-4-isopropylaminobenzoic acid (1.0 g, 4.6 mmol), thionyl chloride (0.38 ml, 5.1 mmol), N-methylpyrrolidin-2-one (0.1 ml) and dichloromethane (20 ml) was stirred together at room temperature for 12 hours. To this solution was added a compound of formula (6) (0.91 g, 2.5 mmol) and the mixture was cooled to 0-5 ° C. After cooling, triethylamine (1.7 ml, 12 mmol) was added, the reaction was warmed to room temperature and stirred for 4 hours. The reaction mixture was filtered to remove precipitated triethylamine hydrochloride, and the organic layer was concentrated under reduced pressure to obtain the title compound as a solid, which was used as such in the next reaction without purification.

Yield: 0.95 g (67%); MS (ES +): 547 (M + 1)

Example 60:

(+) - trans-2- (2-Chloro-4-isopropylaminophenyl) -8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -5,7-dimethoxychromen-4-one

To a solution of n-BuLi (1.6 M in hexane, 2.22 ml, 3.56 mmol) in THF (10 ml), which was maintained at 0 ° C. under a nitrogen atmosphere, hexamethyldisilazane (0.74 ml, 3.56 mmol) was added dropwise and stirred for 15 minutes. To this was added dropwise a solution of the compound of example (59) (0.95 g, 1.78 mmol) in THF (5 ml), maintaining the temperature at 0 ° C. After addition, the reaction was warmed to room temperature and stirred for 2.5 hours. The reaction mixture was acidified with dilute HCl and made basic with 10% sodium bicarbonate to pH 8-9. The aqueous layer was extracted with chloroform (3 × 50 ml). The organic layer was washed with water (25 ml), brine (25 ml) and dried over anhydrous Na ₂ SO ₄ . The organic layer was concentrated under reduced pressure and dried under vacuum to obtain acetic acid 3- {3- [3- (2-chloro-4-isopropylaminophenyl) -3-oxo-propionyl] -2-hydroxy-4,6-dimethoxyphenyl} - 1-methylpyrrolidin-2-yl-methyl ester as a brown solid (0.95 g). This ester was dissolved in conc. HCl (5 ml) and stirred for 3 hours to complete cyclization. At the end of 3 hours, the reaction mixture was basified with solid NaHCO ₃ to pH 8-9. The aqueous layer was extracted with chloroform (3 × 25 ml) and washed with water (10 ml) and brine (10 ml). The organic layer was dried over anhydrous Na ₂ SO ₄ , concentrated under reduced pressure, and dried under vacuum to obtain the compound (+) - trans-2- (2-chloro-4-isopropylaminophenyl) -8- (2-hydroxymethyl-1-methylpyrrolidine- 3-yl) -5,7-dimethoxychromen-4-one as a brown solid, which was taken further without further purification.

Yield: 0.7 g (92.5%)

Example 61:

(+) - trans-2- (2-Chloro-4-isopropylaminophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one

A mixture of the compound of Example (60) (0.7 g, 1.4 mmol), pyridine hydrochloride (0.7 g, 6 mmol) and a catalytic amount of quinoline was heated at 180 ° C. for a period of 2.5 hours. The reaction mixture was diluted with methanol (25 ml) and basified with solid Na ₂ CO ₃ to pH 10. The reaction mixture was filtered and washed with methanol. The organic layer was concentrated and the residue was purified by column chromatography using 0.1% ammonia and 3% methanol in chloroform as an eluent to obtain the compound (+) - trans-2- (2-chloro-4-isopropylaminophenyl) -5,7-dihydroxy- 8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one as a yellow solid.

Yield: 20 mg (3%);

¹ H NMR (CDCl ₃ , 300 MHz): δ 7.38 (d, 1H), 6.63 (d, 1H), 6.51 (dd, 1H), 6.39 (s, 1H), 6, 27 (s, 1H), 4.15 (m, 1H), 3.94 (m, 2H), 3.67 (m, 1H), 3.27 (m, 1H), 2.9 (m, 2H ), 2.65 (s, 3H), 2.43 (m, 2H), 1.25 (d, 6H);

MS (ES +): 459 (M + 1).

Example 62:

(+) - trans-2- (2-Chloro-4-isopropylaminophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one hydrochloride

The compound of formula (61) (20 mg, 0.043 mmol) was suspended in methanol (2 ml) and treated with ethereal HCl and the organic solvent was evaporated to give the salt (+) - trans-2- (2-chloro-4-isopropylaminophenyl) -5, 7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one hydrochloride.

Yield: 20 mg (95%);

¹ H NMR (CD ₃ OD, 300 MHz): δ 7.72 (d, 1H), 7.04 (d, 1H), 6.92 (dd, 1H), 6.58 (s, 1H), 6 , 37 (s, 1H), 4.35 (m, 1H), 3.95 (m, 1H), 3.8 (m, 1H), 3.75 (m, 2H), 3.65 (dd, 1H), 3.40 (m, 1H), 3.0 (s, 3H), 2.53 (m, 1H), 2.32 (m, 1H), 1.29 (d, 6H);

MS (ES-): 493 (M-1).

Example 63:

(+) - trans-2,4-Dibromobenzoic acid 2- (2-acetoxymethyl-1-methylpyrrolidin-3-yl) -6-acetyl-3,5-dimethoxyphenyl ester

A mixture of the compound of example (6) (0.84 g, 2.3 mmol), 2,4-dibromobenzoic acid (0.8 g, 2.8 mmol), DCC (0.98 g, 4.7 mmol) and DMAP (0.3 g, 2.3 mmol) was dissolved in dichloromethane (25 ml) and stirred at room temperature for 12 hours. The reaction mixture was cooled to 0 ° C, the precipitated dicyclohexylurea was filtered and the organic layer was concentrated and the residue was purified by column chromatography with 1% methanol in chloroform and 0.1% ammonia as an eluent to obtain the compound (+) - trans-2,4-dibromo benzoic acid 2- (2-acetoxymethyl-1-methylpyrrolidin-3-yl) -6-acetyl-3,5-dimethoxyphenyl ester.

Yield: 1.14 g (78%);

¹ H NMR (CDCl ₃ ; 300 MHz): δ 8.19 (s, 1H), 7.55 (m, 2H), 6.44 (s, 1H), 4.2 (m, 2H), 3, 91 (s, 6H), 3.60 (m, 1H), 3.18 (m, 1H), 2.84 (m, 1H), 2.60 (s, 3H), 2.48 (s, 3H ), 2.13 (m, 1H), 1.94 (m, 2H), 1.80 (s, 3H);

MS (ES +): 614 (M + 1).

Example 64:

(+) - trans-2- (2,4-dibromophenyl) -8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -5,7-dimethoxychromen-4-one

To a solution of n-BuLi (15%) in hexane, 2.38 ml, 5.6 mmol) in THF (10 ml), which was maintained at 0 ° C in a nitrogen atmosphere, hexamethyldisilazane (1.67 ml, 8 mmol) - added dropwise and stirred for 15 minutes. To this was added dropwise a solution of the compound of Example (63) (1.14 g, 1.8 mmol) in THF (10 ml), maintaining the temperature at 0 ° C. After addition, the reaction was warmed to room temperature and stirred for 2.5 hours. The reaction mixture was acidified with dilute HCl and made basic with 10% sodium bicarbonate to pH 8-9. The aqueous layer was extracted with chloroform (3 × 25 ml). The organic layer was washed with water (25 ml), brine (25 ml) and dried over anhydrous Na ₂ SO ₄ . The organic layer was concentrated under reduced pressure and dried in vacuo to give 3- {3- [3- (2,4-dibromophenyl) -3-oxopropionyl] -2-hydroxy-4,6-dimethoxyphenyl} -1-methylpyrrolidine- 2-yl-methyl ester as a brown solid (1.2 g). This ester was dissolved in conc. HCl (10 ml) and stirred for 3 hours to perform cyclization. At the end of 3 hours, the reaction mixture was basified with solid NaHCO ₃ to pH 10. The aqueous layer was extracted with chloroform (25 × 3 ml) and washed with water (25 ml) and brine (25 ml). The organic layer was dried over anhydrous Na ₂ SO ₄ ), concentrated under reduced pressure, and dried under vacuum. The residue was purified by column chromatography with 3% methanol in chloroform and 0.1% ammonia as an eluent to obtain the compound (+) - trans-2- (2,4-dibromophenyl) -8- (2-hydroxymethyl-1-methylpyrrolidin-3- il) -5,7-dimethoxychromen-4-one as a yellow solid.

Yield: 0.6 g (58%);

¹ H NMR (CDCl ₃ , 300 MHz): δ 7.81 (s, 1H), 7.59 (d, 1H), 7.51 (dd, 1H), 6.48 (s, 1H), 6, 44 (s, 1H), 4.21 (m, 1H), 4.03 (s, 3H), 4.01 (s, 3H), 3.73 (m, 1H), 3.50 (m, 1H ), 3.33 (m, 1H), 2.99 (m, 1H), 2.66 (m, 1H), 2.50 (s, 3H), 2.15 (m, 2H); MS (ES +): 554 (M + 1).

Example 65:

(+) - trans-2- (2,4-dibromophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one

A mixture of the compound of Example (64) (0.6 g, 1.08 mmol), pyridine hydrochloride (0.6 g, 5.19 mmol) and a catalytic amount of quinoline was heated at 180 ° C. for a period of 2.5 hours. The reaction mixture was diluted with methanol (50 ml) and basified with solid Na ₂ CO ₃ to pH 10. The reaction mixture was filtered and washed with methanol. The organic layer was concentrated and the residue was purified by column chromatography using 0.1% ammonia and% methanol in chloroform as an eluent to obtain the title compound as a yellow solid.

Yield: 0.2 g (22%); MS (ES +): 526 (M + 1).

Example 66:

(+) - trans-2- (2,4-dibromophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one hydrochloride

The compound of formula (65) (25 mg, 0.047 mmol) was suspended in methanol (2 ml) and treated with ethereal HCl and the organic solvent was evaporated to give the salt (+) - trans-2- (2,4-dibromophenyl) -5,7- dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one hydrochloride.

Yield: 25 mg (93.5%);

¹ H NMR (CD ₃ OD, 300 MHz): δ 7.95 (m, 1H), 7.73 (m, 2H), 6.52 (s, 1H), 6.38 (s, 1H), 3 90 (m, 1H), 3.85 (m, 2H), 3.60 (m, 2H), 3.4 (m, 1H), 2.97 (s, 3H), 2.53 (m, 1H); 2.28 (m, 1H);

MS (ES-): 560 (M-1) ⁺

Example 67:

(+) - trans-2-bromo-4-chlorobenzoic acid 2- (2-acetoxymethyl-1-methylpyrrolidin-3-yl) -6-acetyl-3,5-dimethoxyphenyl ester

A mixture of the compound of example (6) (1.12 g, 3.2 mmol), 2-bromo-4-chlorobenzoic acid (0.9 g, 3.8 mmol), DCC (1.32 g, 6.4 mmol) and DMAP (0.4 g, 3.2 mmol) was dissolved in dichloromethane (25 ml) and stirred at room temperature for 12 hours. The reaction mixture was cooled to 0 ° C, the precipitated dicyclohexylurea was filtered off and the organic layer was concentrated and the residue was purified by column chromatography with 1% methanol in chloroform and 0.1%) ammonia as an eluent to obtain the title compound (+) - trans-2-bromo-4 -chloro-benzoic acid 2- (2-acetoxymethyl-1-methylpyrrolidin-3-yl) -6-acetyl-3,5-dimethoxyphenyl ester.

Yield: 0.6 g (33%); MS (ES +): 569 (M + 1).

Example 68:

(+) - trans-2- (2-Bromo-4-chlorophenyl) -8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -5,7-dimethoxychromen-4-one

To a solution of n-BuLi (15% in hexane, 0.9 ml, 2 mmol) in THF (10 ml), which was maintained at 0 ° C. under nitrogen, hexamethyldisilazane (0.44 ml, 2 mmol) was added dropwise and stirred for 15 minutes. To this was added dropwise a solution of the compound of Example (67) (0.6 g, 1 mmol) in THF (10 ml), maintaining the temperature at 0 ° C. After addition, the reaction was warmed to room temperature and stirred for 2.5 hours. The reaction mixture was acidified with dilute HCl and made basic with 10% sodium bicarbonate to pH 8-9. The aqueous layer was extracted with chloroform (3 × 25 ml). The organic layer was washed with water (25 ml), brine (25 ml) and dried over anhydrous Na ₂ SO ₄ . The organic layer was concentrated under reduced pressure and dried under vacuum to obtain 3- {3- [3- (2-bromo-4-chlorophenyl) -3-oxopropionyl] -2-hydroxy-4,6-dimethoxyphenyl} -1- methylpyrrolidin-2-yl-methyl ester as a brown solid (1.2 g). This ester was dissolved in conc. HCl (10 ml) and stirred for 3 hours to perform cyclization. At the end of 3 hours, the reaction mixture was basified with solid NaHCO ₃ to pH 8-9. The aqueous layer was extracted with chloroform (25 × 3 ml) and washed with water (25 ml) and brine (25 ml). The organic layer was dried over anhydrous Na ₂ SO ₄ ; concentrated under reduced pressure and dried under vacuum. The residue was purified by column chromatography with 3% methanol in chloroform and 0.1% ammonia as an eluent to obtain the compound (+) - trans-2- (2-bromo-4-chlorophenyl) -8- (2-hydroxymethyl-1-methylpyrrolidine- 3-yl) -5,7-dimethoxychromen-4-one as a yellow solid.

Yield: 0.3 g (53.6%);

MS (ES +): 509 (M + 1).

Example 69:

(+) - trans-2- (2-Bromo-4-chlorophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one

A mixture of the compound of Example (68) (0.3 g, 0.59 mmol), pyridine hydrochloride (0.3 g, 2.6 mmol) and a catalytic amount of quinoline was heated at 180 ° C. for a period of 2.5 hours. The reaction mixture was diluted with methanol (25 ml) and basified with solid Na ₂ CO ₃ to pH 10. The reaction mixture was filtered and washed with methanol. The organic layer was concentrated and the residue was purified by column chromatography using 0.1% ammonia and 3% methanol in chloroform as an eluent to obtain the compound (+) - trans-2- (2-bromo-4-chlorophenyl) -5,7-dihydroxy- 8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one as a yellow solid.

Yield: 0.04 g (14%);

MS (ES +): 481 (M + 1).

Example 70:

(+) - trans-2- (2-Bromo-4-chlorophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one hydrochloride

The compound of formula (69) (34 mg, 0.07 mmol) was suspended in methanol (2 ml) and treated with ethereal HCl and the organic solvent was evaporated to give the salt (+) - trans-2- (2-bromo-4-chlorophenyl) - 5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one hydrochloride.

Yield: 34 mg (93%);

¹ HNMR (CD ₃ OD, 300 MHz): δ 7.98 (m, 1H), 7.8 (m, 2H), 6.5 (s, 1H), 6.37 (s, 1H), 4, 17 (m, 1H), 3.88 (m, 2H), 3.59 (m, 2H), 3.43 (m, 1H), 3.00 (s, 3H), 2.49 (m, 1H ), 2.28 (m, 1H);

MS (ES-): 515 (M-1).

Example 71:

(±) -trans-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -5,7-dimethoxy-2- (2-methoxyphenyl) -chromen-4-one

To a suspension of 50% sodium hydride in mineral oil (1.39 g, 29.1 mmol) in N, N-dimethylformamide (60 ml), maintained in a nitrogen atmosphere and cooled to 0 ° C, (±) -trans-1- [2-Hydroxy-3- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -4,6-dimethoxyphenyl] ethanone (3 g, 9.7 mmol) was added portionwise and stirred for 15 minutes. At the end of 15 minutes, 2-methoxy methyl benzoate (4.8 g, 29 mmol) was added dropwise and the reaction was stirred at 0 ° C. for 1 hour. Excess sodium hydride was destroyed by careful addition of methanol. The reaction mixture was acidified with diluted HCl and made basic with 10% sodium bicarbonate to pH 9. The reaction mixture was concentrated under reduced pressure and dried in vacuo to give 1- [2-hydroxy-3- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) - 4,6-dimethoxyphenyl] -3- (2-methoxyphenyl) propane-1,3-dione as an oil (1.4 g). This (β-diketone was dissolved in conc. HCl (50 ml) and stirred for 3 hours to perform cyclization. At the end of 3 hours, the reaction mixture was basified with solid NaHCO ₃ to pH 10. The aqueous layer was extracted with chloroform (50 × 3 ml) and washed with water (25 ml) and brine (25 ml). The organic layer was dried over anhydrous Na ₂ SO ₄ , concentrated under reduced pressure and dried in vacuo. The residue was purified by column chromatography with 3% methanol in chloroform and 0.1% ammonia as eluent to obtain the compound (±) -trans-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -5,7-dimet hydroxy-2- (2-methoxyphenyl) -chromen-4-one.

Yield: 1.4 g (34%);

¹ H NMR (CDCl ₃ , 300 MHz): δ 7.97 (d, 1H), 7.44 (t, 1H), 7.09 (t, 1H), 7.01 (d, 1H), 6, 91 (s, 1H), 6.43 (s, 1H), 4.27 (m, 1H), 4.00 (s, 3H), 3.96 (s, 3H), 3.90 (s, 3H ), 3.70 (dd, 1H), 3.42 (d, 1H), 3.23 (m, 1H), 2.72 (m, 2H), 2.41 (s, 3H), 2.08 (m, 2H);

MS (ES +): 426 (M + 1).

Example 72:

(±) -trans-5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -2- (2-hydroxyphenyl) -chromen-4-one (compound-1);

(±) -trans-5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -2- (2-methoxyphenyl) -chromen-4-one (compound-2);

(±) -trans-5-hydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -7-methoxy-2- (2-methoxyphenyl) -chromen-4-one (compound-3);

A mixture of the compound of Example (71) (70 mg, 0.165 mmol), pyridine hydrochloride (700 mg, 6.09 mmol) was heated at 180 ° C. for a period of 2.5 hours. The reaction mixture was diluted with methanol (50 ml) and basified with solid Na ₂ CO ₃ to pH 10. The reaction mixture was filtered and washed with methanol. The organic layer was concentrated and the residue was purified by column chromatography using 0.1% ammonia and 3% methanol in chloroform as an eluent to obtain (±) -trans-5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3 compounds -yl) -2- (2-hydroxyphenyl) -chromen-4-one (compound-1), (±) -trans-5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -2- (2-methoxyphenyl) -chromen-4-one (compound-2) and (±) -trans-5-hydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -7-methoxy- 2- (2-methoxyphenyl) -chromen-4-one (compound-3).

(Compound-1): Yield: 17 mg (27%);

¹ H NMR (CDCl ₃ , 300 MHz): δ 7.90 (dd, 1H), 7.36 (m, 1H), 7.00 (s, 1H), 6.96 (m, 2H), 6, 18 (s, 1H), 4.20 (m, 1H), 3.79 (dd, 1H), 3.65 (dd, 1H), 3.55 (m, 1H), 3.45 (m, 2H ), 2.81 (s, 3H); 2.34 (m, 2H);

MS (ES +): 384 (M + 1).

(Compound-2): Yield: 20 mg (30%);

¹ H NMR (CDCl ₃ , 300 MHz): δ 12.77 (s, 1H), 7.82 (dd, 1H), 7.44 (m, 1H), 7.05 (m, 2H), 6, 88 (s, 1H), 6.27 (s, 1H), 4.20 (m, 1H), 3.95 (m, 1H), 3.91 (s, 3H), 3.80 (m, 1H ), 3.30 (m, 2H), 2.84 (m, 1H), 2.67 (s, 3H), 2.50 (m, 1H), 2.12 (m, 1H); MS (ES +): 398 (M + 1).

(Compound-3): Yield: 12 mg (18%);

¹ H NMR (CDCl ₃ , 300 MHz): δ 13.12 (s, 1H), 8.03 (m, 1H), 7.48 (m, 1H), 7.11 (m, 1H), 7, 02 (m, 1H), 6.99 (s, 1H), 6.42 (s, 1H), 4.21 (m, 1H), 3.93 (s, 3H), 3.91 (s, 3H ), 3.58 (m, 2H), 3.11 (m, 1H), 2.70 (m, 2H), 2.39 (s, 3H), 2.05 (m, 2H); MS (ES +): 412 (M + 1).

Example 73:

(±) -trans-5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -2- (2-hydroxyphenyl) -chromen-4-one hydrochloride

Compound-1 of Example 72 (15 mg, 0.039 mmol) was suspended in methanol (2 ml) and treated with ethereal HCl and the organic solvent was evaporated to obtain the salt of (±) -trans-5,7-dihydroxy-8- (2-hydroxymethyl-1 methyl pyrrolidin-3-yl) -2- (2-hydroxyphenyl) -chromen-4-one hydrochloride.

Yield: 14 mg (85%);

¹ H NMR (CD ₃ OD, 300 MHz): δ 7.97 (dd, 1H), 7.38 (m, 1H), 7.11 (s, 1H), 7.00 (m, 2H), 6 34 (s, 1H), 4.42 (m, 1H), 3.90 (m, 2H), 3.79 (m, 1H), 3.63 (dd, 1H), 3.49 (m, 1H), 3.01 (s, 3H), 2.58 (m, 1H), 2.36 (m, 1H);

MS (ES +): 420 (M + 1) ⁺

Example 74:

(+) Acetic acid 3- [2- (2-chlorophenyl) -5,7-dihydroxy-4-oxo-4H-chromen-8-yl] -1-methylpyrrolidin-2-yl-methyl ester

The compound of formula (9) (2 g, 5 mmol) was stirred in dry dichloromethane (40 ml) with acetic anhydride (2.50 ml, 26.4 mmol) and a catalytic amount of dimethylaminopyridine at 25 ° C. After 1 hour, a saturated sodium carbonate solution (30 ml) was added to the reaction mixture and stirred for 15 minutes. The organic layer was separated and the aqueous layer was extracted with CHCl ₃ (3 × 50 ml). The combined organic layer was concentrated and the resulting product was purified using a silica gel column and 3% methanol in CHCl ₃ + 1% NH ₃ liquor as eluent to obtain 3- [2- (2-chlorophenyl) -5.7 (+) acetic acid compound -dihydroxy-4-oxo-4H-chromen-8-yl] -1-methylpyrrolidin-2-yl-methyl ester.

Yield: 1.3 g (60%);

IR (KBr): 3421, 3069, 2953, 1742, 1659 cm ^-1 ;

¹ H NMR (CDCl ₃ , 300 MHz): δ 12.60 (s, 1H), 7.54 (m, 2H), 7.45 (m, 2H), 6.44 (s, 1H), 6, 31 (s, 1H), 4.30 (m, 1H), 4.20 (m, 1H), 4.02 (m, 1H), 3.42 (t, 1H), 3.32 (t, 1H ), 2.77 (m, 1H), 2.62 (s, 3H), 2.43 (m, 1H), 2.00 (m, 1H), 1.90 (s, 3H).

Example 75:

(+) Acetic acid 3- [2- (2-chlorophenyl) -5,7-dihydroxy-4-oxo-4H-chromen-8-yl] -1-methylpyrrolidin-2-yl-methyl ester hydrochloride

The compound of formula (74) (0.2 g, 0.38 mmol) was suspended in IPA (5 ml) and 3.5% HCl (25 ml) was added. The suspension was heated until a clear solution was obtained. The solution was cooled and the solid was filtered to give the title compound (+) acetic acid 3- [2- (2-chlorophenyl) -5,7-dihydroxy-4-oxo-4H-chromen-8-yl] -1-methylpyrrolidin-2 methyl yl hydrochloride ester.

Yield: 1.00 g (93%); MS (ES +): m / z 444 (M + 1), free base.

Example 76:

(+) - trans-Acetic acid 8- (2-acetoxymethylpyrrolidin-3-yl) -2- (2-chlorophenyl) -5-hydroxy-4-oxo-4H-chromene-7-yl ester

A solution of the compound of example (10) (10 g, 22 mmol) in pyridine (100 ml) was cooled to 0 ° C. Acetyl chloride (5.7 ml, 80 mmol) was added dropwise and the reaction mixture was warmed to room temperature, followed by heating under reflux for 12 hours. The mixture was concentrated under reduced pressure and the residue was purified by column chromatography using 5% methanol in chloroform as an eluent to obtain the title compound (+) - trans-acetic acid 8- (2-acetoxymethylpyrrolidin-3-yl) -2- (2-chlorophenyl) -5-hydroxy-4-oxo-4H-chromen-7-yl ester.

Yield: 5 g (45%);

¹ H NMR (CDCl ₃ , 300 MHz): δ 7.54 (d, 1H), 7.44 (m, 3H), 6.53 (s, 1H), 6.44 (s, 1H), 4, 35 (m, 1H), 4.17 (m, 2H), 3.48 (m, 1H), 3.35 (m, 1H), 2.79 (m, 1H), 2.62 (s, 3H ), 2.44 (m, 1H), 2.42 (s, 3H), 1.92 (m, 1H), 2.0 (s, 3H);

MS (ES +): m / z 486 (M + 1).

Example 77:

(+) - trans-Benzoic acid 8- (2-benzoyloxymethylpyrrolidin-3-yl) -2- (2-chloro phenyl) -5-hydroxy-4-oxo-4H-chromene-7-yl ester

A solution of the compound of example (10) (0.5 g, 1.1 mmol) in pyridine (3 ml) was cooled to 0 ° C. Benzoyl chloride (1 ml, 8.62 mmol) was added dropwise and the reaction mixture was warmed to room temperature, followed by heating under reflux for 12 hours. The mixture was concentrated under reduced pressure and the residue was purified by column chromatography using 5% methanol in chloroform as an eluent to obtain the compound (+) - trans-benzoic acid 8- (2-benzoyloxymethylpyrrolidin-3-yl) -2- (2-chlorophenyl) - 5-hydroxy-4-oxo-4H-chromen-7-yl ester.

Yield: 0.2 g (28%);

¹ H NMR (CDCl ₃ , 300 MHz): δ 8.25 (m, 1H), 8.09 (m, 1H), 7.91 (m, 1H), 7.50 (m, 11 H), 6 68 (s, 1H), 6.34 (s, 1H), 4.55 (m, 2H), 4.32 (dd, 1H), 3.69 (m, 1H), 3.40 (m, 1H), 2.90 (m, 1H), 2.72 (s, 3H), 2.58 (m, 1H), 2.12 (m, 1H);

MS (ES +): m / z 610 (M + 1).

Example 78:

(+) - trans-Benzoic acid 8- (2-benzoyloxymethylpyrrolidin-3-yl) -2- (2-chloro phenyl) -5-hydroxy-4-oxo-4H-chromene-7-yl ester hydrochloride

The compound of formula (77) (0.2 g, 0.328 mmol) was suspended in methanol (2 ml) and treated with ethereal HCl and the organic solvent was evaporated to give the salt of (+) - trans-benzoic acid 8- (2-benzoyloxymethylpyrrolidin-3-yl ) -2- (2-chlorophenyl) -5-hydroxy-4-oxo-4H-chromen-7-yl hydrochloride ester.

Yield: 0.2 g (94.5%);

¹ H NMR (CD ₃ OD, 300 MHz): δ 8.2 (m, 1H), 8.02 (m, 1H), 7.81 (m, 1H), 7.49 (m, 11H), 6 85 (s, 1H), 6.29 (s, 1H), 4.69 (m, 1H), 4.5 (m, 3H), 3.89 (m, 1H), 3.62 (m, 1H), 3.12 (s, 3H), 2.74 (m, 1H), 2.48 (m, 1H);

MS (ES +): m / z 610 (M + 1), free base.

Example 79:

(+) - trais-octanoic acid 2- (2-chlorophenyl) -5-hydroxy-8- (1-methyl-2-octanoyloxymethylpyrrolidin-3-yl) -4-oxo-4H-chromene-7-yl ester

A solution of compound (10) (0.5 g, 1.2 mmol) in pyridine (3 ml) was cooled to 0 ° C. Octanoyl chloride obtained by heating octanoic acid under reflux (0.41 ml, 3.12 mmol) in thionyl chloride (1 ml) over 30 minutes, followed by evaporation to dryness, was added dropwise and the reaction mixture was heated to room temperature, followed by heating with reflux for 12 hours. The mixture was concentrated under reduced pressure and the residue was purified by column chromatography using 5% methanol in chloroform as an eluent to obtain the title compound.

Yield: 0.2 g (30.5%); ¹ H NMR (CDCl ₃ , 300 MHz): δ 12.60 (s, 1H), 7.55 (m, 4H), 6.44 (s, 1H), 6.31 (s, 1H), 4, 31 (m, 1H), 4.18 (m, 1H), 4.03 (m, 1H), 3.42 (m, 1H), 3.32 (m, 1H), 2.76 (m, 1H) ), 2.62 (s, 3H), 2.45 (m, 1H), 2.34 (t, 2H), 2.17 (m, 2H), 1.95 (m, 1H), 1.63 (m, 2H), 1.48 (m, 2H), 1.27 (m, 16H), 0.86 (m, 6H).

Example 80:

(+) - trans-acetic acid 3- [2- (2-chlorophenyl) -5-hydroxy-7-methoxy-4-oxo-4H-chromen-8-yl] -1-methylpyrrolidin-2-yl-methyl complex ether

To a solution of the compound of formula (8) (2.0 g, 4.65 mmol) and dimethylaminopyridine (10 mg) in dichloromethane (20 ml), acetic anhydride (0.48 ml, 5.07 mmol) was added dropwise and the reaction mixture was stirred for 15 minutes. At the end of 15 minutes, the reaction mixture was diluted with chloroform (20 ml), the organic layer was washed with 10% NaHCO ₃ , water (10 ml) and brine (10 ml). The organic layer was dried over anhydrous Na ₂ SO ₄ and concentrated under reduced pressure. The residue was dissolved in dichloromethane (20 ml), followed by the addition of boron trifluoride diethyl ether (1.72 ml, 13.9 mmol) and dimethyl sulfide (1.1 ml, 14 mmol) and stirred at room temperature for 1 hour. The reaction mixture was poured onto ice, basified with 10% Na ₂ CO ₃ and extracted with chloroform (2 × 50 ml). The organic layers were combined, washed with water (10 ml), brine (10 ml), dried over anhydrous Na ₂ SO ₄ and concentrated under reduced pressure. The residue was purified by column chromatography using 20% petroleum ether in chloroform and 0.1% ammonia as an eluent to obtain 3- [2- (2-chlorophenyl) -5-hydroxy-7-methoxy- (+) - trans-acetic acid compound 4-oxo-4H-chromen-8-yl] -1-methylpyrrolidin-2-yl-methyl ester.

Yield: 1.22 g (58%);

¹ H NMR (CDCl ₃ , 300 MHz): δ 12.88 (s, 1H), 7.50 (m, 4H), 6.47 (s, 1H), 6.44 (s, 1H), 4, 04 (m, 2H), 3.95 (m, 1H), 3.90 (s, 3H), 3.13 (m, 1H), 2.74 (m, 1H), 2.50 (m, 1H ), 2.36 (s, 3H), 2.13 (m, 1H), 1.95 (m, 1H), 1.72 (s, 3H);

MS (ES +): m / z 458 (M + 1).

Example 81:

(+) - trans-2- (2-Chlorophenyl) -5-hydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -7-methoxychromen-4-one

Method A

The compound of formula (80) (1.4 g, 3.0 mmol) was dissolved in methanol (6 ml), to which a 2% aqueous NaOH solution (6 ml) was added, and the mixture was heated with stirring at 50-60 ° C for 1 hour At the end of this period, the mixture was acidified with dilute HCl, followed by basification with 10% Na ₂ CO ₃ solution to pH 9. The mixture was extracted with EtOAc (3 × 25 ml) and the organic layers were washed with water (10 ml), brine (10 ml), dried over anhydrous Na ₂ SO ₄ was concentrated under reduced pressure and the residue was purified by column chromatography using 1% isopropanol in chloroform and 0.1% ammonia as eluent to obtain compound (+) - trans-2- (2-chlorophenyl) -5- hydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -7-methoxychromen-4-one. Yield: 1.2 g (97%);

IR (KBr): 3446, 1656, 1608 cm ^-1 ;

¹ H NMR (CDCl ₃ , 300 MHz): δ 12.90 (s, 1H), 7.74 (m, 1H), 7.48 (m, 3H), 6.57 (s, 1H), 6, 44 (s, 1H), 4.11 (m, 1H), 3.91 (s, 3H), 3.63 (dd, 1H), 3.36 (d, 1H), 3.14 (m, 1H) ), 2.72 (m, 1H), 2.55 (m, 1H), 2.33 (s, 3H), 2.01 (m, 2H);

MS (ES +): m / z 416 (M + 1) ⁺ .

Method B

The compound of formula (8) (0.1 g, 0.23 mmol) was treated with HBr (5 ml, 47%) at 100 ° C. for 3.5 hours. The reaction mixture was poured onto ice, made basic with a solution of Na ₂ CO ₃ (10%, 40 ml), extracted with ethyl acetate and concentrated. The resulting product was purified by column chromatography and 5% methanol + 1% solution of NH ₃ in CHCl ₃ as eluent to obtain the title compound.

Yield: 0.085 g (84%); ¹ H NMR (CDCl ₃ , 300 MHz): δ 12.90 (s, 1H), 7.74 (m, 1H), 7.48 (m, 3H), 6.57 (s, 1H), 6, 44 (s, 1H), 4.11 (m, 1H), 3.91 (s, 3H), 3.63 (dd, 1H), 3.36 (d, 1H), 3.14 (m, 1H ), 2.72 (m, 1H), 2.55 (m, 1H), 2.33 (s, 3H), 2.01 (m, 2H); MS (ES +): m / z 416 (M + 1).

Example 82:

(+) - trans-2- (2-Chlorophenyl) -5-hydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -7-methoxychromen-4-one hydrochloride

The compound of formula (81) (50 mg, 0.11 mol) was suspended in methanol (2 ml), treated with ethereal HCl and the organic solvent was evaporated to give the salt (+) - trans-2- (2-chlorophenyl) -5-hydroxy- 8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -7-methoxychromen-4-one hydrochloride.

Yield: 53 mg (100%);

IR (KBr): 3297, 1663, 1608, 1581 cm ^-1 ;

¹ H NMR (CD ₃ OD, 300 MHz): δ 8.12 (d, 1H), 7.91 (m, 3H), 6.96 (s, 1H), 6.89 (s, 1H), 4 62 (m, 1H), 4.34 (s, 3H), 4.17 (m, 2H), 4.03 (m, 1H), 3.90 (m, 1H), 3.71 (m, 1H), 3.31 (s, 3H), 2.75 (m, 1H), 2.62 (m, 1H); MS (ES +): m / z 416 (M-36.5);

[α] _D ²⁵ : + 15.32 ° (c = 0.27, methanol).

Example 83:

(+) - trans-2- (2-Chlorophenyl) -5-hydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -7-methoxychromen-4-one gluconate

The compound of formula (81) (50 mg, 0.12 mmol) was suspended in methanol (1 ml) and water (1 ml) and stirred. To this was added gluconic acid (24 mg, 0.12 mmol), stirred and the solution was evaporated dry to obtain the salt (+) - trax-2- (2-chlorophenyl) -5-hydroxy-8- (2-hydroxymethyl-1- methylpyrrolidin-3-yl) -7-methoxychromen-4-one gluconate.

Yield: 74 mg (100%); IR (KBr): 3445, 1654, 1609 cm ^-1 ; ¹ H NMR (CD ₃ OD, 300 MHz): δ 7.80 (d, 1H), 7.59 (m, 3H), 6.64 (s, 1H), 6.57 (s, 1H), 4 , 27 (m, 1H), 4.07 (m, 2H), 4.02 (s, 3H), 3.76 (m, 7H), 2.92 (s, 3H), 2.33 (m, 2H); [α] _D ²⁵ : + 18.17 ° (c = 0.73, methanol)

Example 84:

(+) - trans-Acetic acid-3- [2- (2-chlorophenyl) -5,7-dihydroxy-4-oxo-4H-chromen-8-yl] -1-methylpyrrolidin-2-yl-methyl ester

To a solution of the compound of Example (10) (1 g, 2.28 mmol) and triethylamine (1.6 ml, 11.4 mmol) in methylene chloride (10 ml) was added acetic anhydride (0.7 ml, 7.4 mmol) and stirred for 30 minutes at ambient temperature. The reaction mixture was quenched with saturated sodium carbonate (10 ml) and stirred for 30 minutes. The organic layer was separated, washed with aqueous sodium carbonate (10 ml), water (10 ml), dried over anhydrous sodium sulfate and evaporated under reduced pressure to obtain compounds (+) - trans-acetic acid-3- [2- (2-chlorophenyl) -5,7-dihydroxy-4-oxo-4H-chromen-8-yl] -1-methylpyrrolidin-2-yl-methyl complex ether, which was used as such without further purification.

Yield: 1.1 g (99%);

¹ H NMR (CDCl ₃ , 300 MHz) δ 12.59 (s, 1H), 7.45 (m, 4H), 6.44 (s, 1H), 6.31 (s, 1H), 4.32 (m, 1H), 4.19 (m, 1H), 4.05 (m, 1H), 3.44 (m, 1H), 3.30 (m, 1H), 2.79 (m, 1H) 2.62 (s, 3H); 2.42 (m, 1H); 1.98 (m, 1H); 1.92 (s, 3H);

MS (ES +): 444 (M + 1).

Example 85:

(+) - trans-acetic acid 3- [2- (2-chlorophenyl) -5-hydroxy-7- (2-methoxyethoxy methoxy) -4-oxo-4H-chromen-8-yl] -1-methylpyrrolidin-2 methyl ester

The compound of formula (84) ((1.11 g, 2.50 mmol) was dissolved in dry DMF (15 ml) under nitrogen and cooled to 0 ° C. Sodium hydride (50%, 0.14 g, 2.9 mmol) ) was added and the mixture was stirred for 30 minutes A solution of 2-methoxyethoxymethyl chloride (0.33 ml, 2.9 mmol) in dry DMF was added dropwise and the reaction was maintained at 10 ° C. overnight.The reaction was poured into ice water (10 ml), acidified with acetic acid to pH 5, made basic with aqueous Na ₂ CO ₃ (pH 9) and extracted with ethyl acetate (2 × 25 ml). The organic layer was separated, washed with water (15 ml), brine (15 ml), dried over anhydrous sodium sulfate and concentrated under reduced pressure to dryness to obtain the compound (+) - trans-acetic acid-3- [2- (2-chlorophenyl) -5-hydroxy-7- (2-methoxyethoxymethoxy) -4-oxo-4H-chromene -8-yl] -1-methylpyrrolidin-2-yl-methyl ester.

Yield: 1.1 g (92%);

¹ H NMR (CDCl ₃ , 300 MHz): δ 12.78 (s, 1H), 7.56 (m, 4H), 6.68 (s, 1H), 6.48 (s, 1H), 5, 39 (s, 2H), 4.20 (m, 2H), 4.05 (m, 1H), 3.83 (m, 2H), 3.58 (m, 2H), 3.38 (s, 3H ), 2.56 (m, 3H), 2.16 (s, 3H), 1.81 (s, 3H), 1.62 (m, 2H);

MS (ES +): 532 (M + 1).

Example 86:

(+) - trans-2- (2-Chlorophenyl) -5-hydroxy-8- (2-hydroxymethyl) -1-methylpyrrolidin-3-yl) -7- (2-methoxyethoxymethoxy) -chromen-4-one

The compound of formula (85) (1.1 g, 2.07 mmol) was dissolved in 15 ml of a 1% sodium hydroxide solution in a 1: 1 mixture of methanol-water and stirred at room temperature for 1 hour. At the end of 1 hour, the reaction mixture was diluted with ice water, acidified with acetic acid (pH 5), alkalized with aqueous Na ₂ CO ₃ (pH 9) and extracted with ethyl acetate (2 × 25 ml). The organic layer was separated, washed with water (15 ml), brine (15 ml), dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by column chromatography using 1% isopropanol in chloroform and 0.1% ammonia as an eluent to obtain the title compound (+) - trans-2- (2-chlorophenyl) -5-hydroxy-8- (2-hydroxymethyl) -1 methyl pyrrolidin-3-yl) -7- (2-methoxyethoxymethoxy) -chromen-4-one.

Yield: 0.43 g (38.7%);

¹ H NMR (CDCl ₃ , 300 MHz): δ 12.84 (s, 1H), 7.73 (d, 1H), 7.50 (m, 3H), 6.62 (s, 1H), 6, 57 (s, 1H), 5.41 (s, 2H), 4.30 (q, 1H), 3.85 (m, 3H), 3.59 (m, 4H), 3.37 (m, 1H ), 3.35 (s, 3H), 2.96 (m, 1H), 2.72 (s, 3H), 2.39 (m, 1H), 2.24 (m, 1H); MS (ES +): 490 (M + 1).

Example 87:

(+) - trans-2- (2-Chlorophenyl) -5-hydroxy-8- (2-hydroxymethyl) -1-methylpyrrolidin-3-yl) -7- (2-methoxyethoxymethoxy) -chromen-4-one gluconate

To a solution of the compound of Example (86) (0.55 g, 1.12 mmol) in methanol (5 ml) was added (+) - gluconic acid (0.22 g, 1.12 mmol) and the mixture was stirred at room temperature for 30 minutes. The solvent was removed under reduced pressure to obtain the salt of (+) - trans-2- (2-chlorophenyl) -5-hydroxy-8- (2-hydroxymethyl) -1-methylpyrrolidin-3-yl) -7- (2-methoxyethoxymethoxy) -chrome-4-one gluconate.

Yield: 0.77 g (100%);

IR (KBr): 3385, 1656, 1606 cm ^-1 ;

¹ H NMR (CD ₃ OD, 300 MHz): δ 7.62 (d, 1H), 7.54 (m, 3H), 6.74 (s, 1H), 6.57 (s, 1H), 5 49 (s, 2H), 4.29 (m, 1H), 4.07 (s, 2H), 3.76 (m, 13H), 3.34 (s, 3H), 2.90 (s , 3H); 2.38 (m, 2H);

MS (ES +): 490 (M + 1) ⁺ , free base;

[α] _D ²⁵ : + 15.33 ° (c = 0.6, methanol).

Example 88:

(+) - trans-2- (2-chlorophenyl) -8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -5,7-dimethoxychromen-4-one hydrochloride

The compound of formula (8) (50 mg, 0.116 mmol) was suspended in methanol (2 ml) and stirred. To this was added ethereal HCl, the solution was stirred and evaporated to give (+) - trans-2- (2-chlorophenyl) -8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -5,7-dimethoxychromene salt -one hydrochloride.

Yield: 50 mg (93%);

IR (KBr): 3421, 1647, 1599, 1471 cm ^-1 ;

¹ H NMR (CD ₃ OD, 300 MHz): δ 7.95 (d, 1H), 7.56 (m, 3H), 6.76 (s, 1H), 6.47 (s, 1H), 4 36 (m, 1H), 3.95 (s, 3H), 3.85 (s, 3H), 3.82 (m, 2H), 3.68 (m, 2H), 3.35 (m, 1H), 2.99 (s, 3H); 2.36 (m, 2H);

MS (ES +): m / z 430 (M + 1), free base;

[α] _D ²⁵ : + 3.76 ° (c = 0.28, methanol)

Example 89:

(+) - trans-2- (2-Chlorophenyl) -8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -5,7-dimethoxychromen-4-one citrate

The compound of formula (8) (50 mg, 0.116 mmol) was suspended in methanol (2 ml) and stirred. To this was added citric acid (23 mg, 0.12 mmol), the solution was stirred and evaporated to obtain the title salt (+) - trans-2- (2-chlorophenyl) -8- (2-hydroxymethyl-1-methylpyrrolidin-3- il) -5,7-dimethoxychromen-4-one citrate.

Yield: 73 mg (100%);

¹ H NMR (CD ₃ OD, 300 MHz): δ 7.79 (d, 1H), 7.57 (m, 3H), 6.77 (s, 1H), 6.48 (s, 1H), 4 35 (m, 1H), 4.10 (s, 3H), 4.02 (s, 3H), 3.98 (m, 2H), 3.69 (m, 2H), 3.31 (m, 1H), 3.01 (s, 3H), 2.76 (m, 4H), 2.29 (m, 2H).

Example 90:

(+) - trans-2- (2-Chlorophenyl) -8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -5,7-dimethoxychromen-4-one tartrate

The compound of formula (8) (50 mg, 0.116 mmol) was suspended in methanol (2 ml) and stirred. Tartaric acid (17 mg, 0.113 mmol) was added to this, the solution was stirred and evaporated to give the salt (+) - trans-2- (2-chlorophenyl) -8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) - 5,7-dimethoxychromen-4-one tartrate.

Yield: 7 mg (100%);

¹ H NMR (CD ₃ OD, 300 MHz): δ 7.76 (d, 1H), 7.57 (m, 3H), 6.75 (s, 1H), 6.46 (s, 1H), 4 22 (m, 1H), 4.08 (s, 3H), 4.01 (s, 3H), 3.77 (m, 1H), 3.49 (m, 3H), 3.31 (m, 2H), 3.05 (m, 1H), 2.78 (s, 3H), 2.26 (m, 2H);

MS (ES +): m / z 430 (M), free base;

[α] _D ²⁵ : + 5.35 ° (c = 0.21, methanol)

Example 91:

(+) - trans-2- (2-Chlorophenyl) -8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -5,7-dimethoxychromen-4-one glutamate

The compound of formula (8) (50 mg, 0.116 mmol) was suspended in methanol (2 ml) and stirred. Glutamic acid (17 mg, 0.115 mmol) was added to this, the solution was stirred and evaporated to give the salt (+) - trans-2- (2-chlorophenyl) -8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) - 5,7-dimethoxychromen-4-one glutamate.

Yield: 67 mg (100%);

¹ H NMR (CD ₃ OD, 300 MHz): δ 7.79 (d, 1H), 7.57 (m, 3H), 6.77 (s, 1H), 6.48 (s, 1H), 4 41 (m, 4H), 4.10 (s, 3H), 4.02 (s, 3H), 3.98 (m, 1H), 3.85 (m, 2H), 3.69 (m, 4H), 2.95 (s, 3H), 2.38 (m, 2H).

Example 92:

(+) - trans-2- (2-chlorophenyl) -8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -5,7-dimethoxychromen-4-one maleate

The compound of formula (8) (50 mg, 0.116 mmol) was suspended in methanol (2 ml) and stirred. To this was added maleic acid (14 mg, 0.116 mmol), the solution was stirred and evaporated to give the salt (+) - trans-2- (2-chlorophenyl) -8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) - 5,7-dimethoxychromen-4-one maleate.

Yield: 5 mg (78%);

¹ H NMR (CD ₃ OD, 300 MHz): δ 7.78 (d, 1H), 7.62 (d, 1H), 7.56 (t, 1H), 7.49 (t, 1H), 6 76 (s, 1H), 6.47 (s, 1H), 6.25 (s, 2H), 4.35 (m, 1H), 4.09 (s, 3H), 4.01 (s, 3H), 3.83 (m, 2H), 3.69 (m, 1H), 3.56 (m, 1H), 3.32 (m, 1H), 2.95 (s, 3H), 2, 41 (m, 1H); 2.28 (m, 1H).

Example 93:

(+) - trans-2- (2-Chlorophenyl) -8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -5,7-dimethoxychromen-4-one nitrate

The compound of formula (8) (50 mg, 0.116 mmol) was suspended in methanol (2 ml) and stirred. To this was added nitric acid (7.1 mg, 0.113 mmol), the solution was stirred and evaporated to obtain the title salt of (+) - trans-2- (2-chlorophenyl) -8- (2-hydroxymethyl-1-methylpyrrolidin-3- il) -5,7-dimethoxychromen-4-one nitrate.

Yield: 57 mg (99%);

¹ H NMR (CD ₃ OD, 300 MHz): δ 7.79 (d, 1H), 7.53 (m, 3H), 6.77 (s, 1H), 6.49 (s, 1H), 4 35 (m, 1H), 4.10 (s, 3H), 4.02 (s, 3H), 3.86 (m, 2H), 3.74 (m, 1H), 3.57 (m, 1H), 3.39 (m, 1H), 2.96 (s, 3H), 2.43 (m, 1H), 2.32 (m, 1H).

Example 94:

(+) - trans-2- (2-Chlorophenyl) -8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -5,7-dimethoxychromen-4-one acetate

The compound of formula (8) (50 mg, 0.116 mmol) was suspended in methanol (2 ml) and stirred. Acetic acid (7 mg, 0.11 mmol) was added to this, the solution was stirred and evaporated to give the title salt (+) - trans-2- (2-chlorophenyl) -8- (2-hydroxymethyl-1-methylpyrrolidin-3- il) -5,7-dimethoxychromen-4-one acetate.

Yield: 56 mg (100%);

¹ H NMR (CD ₃ OD, 300 MHz): δ 7.79 (d, 1H), 7.57 (m, 3H), 6.77 (s, 1H), 6.48 (s, 1H), 4 35 (m, 1H), 4.17 (s, 3H), 4.02 (s, 3H), 3.98 (m, 1H), 3.86 (m, 2H), 3.70 (m, 1H), 3.57 (m, 1H), 3.31 (s, 3H), 2.95 (s, 3H), 2.43 (m, 1H), 2.30 (m, 1H); [α] _D ²⁵ : + 2.85 ° (c = 0.14, methanol).

Example 95:

(+) - trans-2- (2-Chlorophenyl) -8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -5,7-dimethoxychromen-4-one gluconate

The compound of formula (8) (50 mg, 0.116 mmol) was suspended in methanol (2 ml) and stirred. To this was added gluconic acid (21 mg, 0.11 mmol), the solution was stirred and evaporated to obtain the salt (+) - trans-2- (2-chlorophenyl) -8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl ) -5,7-dimethoxychromen-4-one gluconate.

Yield: 71 mg (100%); IR (KBr): 3445.1648, 1560 cm ^-1 ;

¹ H NMR (CD ₃ OD, 300 MHz): δ 7.61 (d, 1H), 7.59 (m, 3H), 6.76 (s, 1H), 6.73 (s, 1H), 4 75 (m, 2H), 4.09 (s, 3H), 3.82 (s, 3H), 3.70 (m, 9H), 3.30 (m, 1H), 2.89 (s, 3H), 2.34 (m, 2H);

MS (ES +): m / z 430 (M + 1), free base;

[α] _D ²⁵ : + 2.85 ° (c = 0.14, methanol);

Study: (C ₂₉ H ₃₆ ClN ₁₂ O · H ₂ O) C 54.31 (54.08), H 5.90 (5.94), N 2.49 (2.17), Cl 5.62 ( 5.34), water 3.30 (2.79).

Example 96:

(-) - trans-2- (2-chlorophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one hydrochloride

(This compound is the (-) - trans enantiomer of the compound illustrated in Example 10. (-) - trans Enantiomer was obtained to compare its activity with its corresponding (+) - trans enantiomer.).

(A) (+) - trans- [1-Methyl-3- (2,4,6-trimethoxyphenyl) pyrrolidin-2-yl] methanol

(+) - DBTA (112.7 g, 314.0 mmol) was added to the compound of formula (4) (88.5 g, 314 mmol), followed by the addition of methanol (600 ml). The mixture was heated under reflux for 10 minutes, slowly stirred at room temperature for 3 hours, the crystallized salt was filtered and dried.

Yield: 80.5 g (40%);

[α] _D ²⁵ = + 82.11 ° (c = 0.7, methanol).

The salt was mixed with a 10% aqueous solution of Na ₂ CO ₃ (255 ml) and EtOAc (200x3 ml) to obtain the free base in an EtOAc layer. The EtOAc layer was concentrated to obtain the title compound.

Yield: 30 g (98.5%); [α] _D ²⁵ = + 20.2 ° (c = 0.7, methanol); ¹ H NMR (CDCl ₃ , 300 MHz): δ 6.13 (s, 2H), 3.90 (m, 1H), 3.79 (s, 9H), 3.57 (dd, 1H), 3, 38 (d, 1H), 3.13 (m, 1H), 2.69 (m, 1H), 2.47 (m, 1H), 2.34 (s, 3H), 2.00 (m, 1H ), 1.93 (m, 1H).

(B) (+) - trans-acetic acid 3- (3-acetyl-2-hydroxy-4,6-dimethoxyphenyl) -1-methylpyrrolidin-2-yl methyl ester

BF ₃ etherate (36.8 g, 259 mmol) was added dropwise with stirring at 0 ° C. under N ₂ to a solution of the compound of formula ((96- (A)) (14.6 g, 51.9 mmol) in acetic anhydride (24.26 ml, 26.2 g, 256 mmol). The reaction mixture was stirred at room temperature for 2 hours. It was poured onto crushed ice (1 kg), made alkaline using a saturated aqueous solution of Na ₂ CO ₃ and extracted using EtOAc (3 × 200 ml) The organic extract was washed with brine, dried (anhydrous Na ₂ SO ₄ ) and concentrated to obtain the title compound.

Yield: 17.8 g (98%); ¹ H NMR (CDCl ₃ , 300 MHz): δ 6.1 (s, 1H), 4.22 (m, 2H), 4.15 (m, 1H), 3.94 (s, 3H), 3, 92 (s, 3H), 3.82 (m, 1H), 3.57 (m, 1H), 3.23 (m, 1H), 2.85 (s, 3H), 2.64 (s, 3H) ), 2.28 (m, 2H), 2.01 (s, 3H).

(C) (+) - trans-1- [2-Hydroxy-3- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -4,6-dimethoxyphenyl) ethanone

To a solution of the compound of formula (96- (B)) (17.8 g, 50.7 mmol) in methanol (40 ml) was added with stirring, at room temperature, a 10% aqueous solution of NaOH (40 ml). The temperature of the reaction mixture was raised to 50 ° C. over 45 minutes. It was cooled to room temperature, acidified using conc. HCl, and concentrated to remove methanol. It was alkalinized using a saturated aqueous solution of Na ₂ CO ₃ . The precipitated title compound was filtered off, washed with water and dried.

Yield: 10.2 g (65%); mp: 173-174 ° C; [α] _D ²⁵ = + 4.12 ° (c = 0.7, methanol);

¹ H NMR (CDCl ₃ , 300 MHz): δ 5.96 (s, 1H), 3.93 (m, 1H), 3.91 (s 3H), 3.90 (s, 3H), 3.58 (dd, 1H), 3.42 (d, 1H), 3.23 (m, 1H), 2.80 (m, 1H), 2.63 (s, 3H), 2.62 (m, 1H) 2.40 (s, 3H); 2.00 (m, 2H); MS (ES +): m / z 310 (M + 1).

(D) (-) - trans-2- (2-Chlorophenyl) -8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -5,7-dimethoxychromen-4-one

Sodium hydride (50%, 0.54 g, 11.25 mmol) was added portionwise to a solution of the compound of formula (96- (C)) (0.7 g, 2.2 mmol) in dry DMF (15 ml) at 0 ° C, in a nitrogen atmosphere and with stirring. After 10 minutes, methyl 2-chlorobenzoate (1.15 g, 6.75 mmol) was added. The reaction mixture was stirred at 25 ° C for 2 hours. Methanol was carefully added at below 20 ° C. The reaction mixture was poured onto crushed ice (300 g), acidified with 1: 1 HCl (pH 2) and extracted using EtOAc (2 × 100 ml). The aqueous layer was alkalinized using saturated Na ₂ CO ₃ (pH 10) and extracted using CHCl ₃ (3 × 200 ml). The organic layer was dried (anhydrous Na ₂ SO ₄ ) and concentrated. To the residue was added conc. HCl (25 ml) and stirred at room temperature for 2 hours. The reaction mixture was poured onto crushed ice (300 g) and made basic using a saturated aqueous solution of Na ₂ CO ₃ . The mixture was extracted using CHCl ₃ (3 × 200 ml). The organic extract was washed with water, dried (anhydrous Na ₂ SO ₄ ) and concentrated to obtain the title compound.

Yield: 0.625 g (65%); mp: 95-97 ° C; IR (KBr): 3431, 1648, 1598, 1571 cm ^-1 ; [α] _D ²⁵ = -4.95 ° (c = 0.7, methanol); ¹ H NMR (CDCl ₃ , 300 MHz): δ 7.70 (dd, 1H), 7.68 (m, 3H), 6.50 (s, 1H), 6.44 (s, 1H), 4, 20 (m, 1H), 4.00 (s, 3H), 3.96 (s, 3H), 3.66 (dd, 1H), 3.37 (d, 1H), 3.15 (m, 1H ), 2.77 (d, 1H), 2.54 (m, 1H), 2.33 (s, 3H), 2.05 (m, 2H); MS (ES +): m / z 430 (M + 1).

(E) (-) - trans-2- (2-Chlorophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one

Molten pyridine hydrochloride (2 g, 17.3 mmol) was added to the compound of formula (96- (D)) (0.2 g, 0.46 mmol) and heated at 180 ° C. for 1.5 hours. The reaction mixture was cooled to 25 ° C., diluted with MeOH (10 ml) and made basic using Na ₂ CO ₃ to pH 10. The mixture was filtered and the organic layer was concentrated. The residue was suspended in water (5 ml), stirred for 30 minutes, filtered and dried to obtain the title compound.

Yield: 0.125 g (67.6%); mp: 228-230 ° C; IR (KBr): 3422, 3135, 1664, 1623, 1559 cm ^-1 ; MS (ES +): m / z 402 (M + 1); Study: C ₂₁ H ₂₀ ClNO ₅ · 0.5 H ₂ O C, 61.81 (61.33); H, 4.62 (5.00); N, 3.75 (3.33); C1, 8.47 (8.45).

(F) (-) - trans-2- (2-Chlorophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one hydrochloride

The compound of formula (96- (E)) (4.09 g, 10.18 mmol) was suspended in IPA (5 ml) and 3.5% HCl (25 ml) was added. The suspension was heated until a clear solution was obtained. The solution was cooled and the solid was filtered to obtain the title compound.

Yield: 4.47 g (99%); mp: 180-184 ° C; [α] _D ²⁵ = -20.2 ° (c = 0.4, methanol);

¹ H NMR (CD ₃ OD, 300 MHz): δ 7.80 (d, 1H), 7.60 (m, 3H), 6.54 (s, 1H), 6.37 (s, 1H), 4 23 (m, 1H), 3.89 (m, 2H), 3.63 (m, 1H), 3.59 (dd, 1H), 3.38 (m, 1H), 2.97 (s, 3H); 2.51 (m, 1H); 2.30 (m, 1H); MS (ES +): m / z 402 (M-36.5); Study: C ₂₁ H ₂₁ Cl ₂ NO ₅ C, 57.2 (57.55); H, 4.92 (4.83); N, 2.98 (3.20); Cl, 15.74 (16.18).

Example 97:

(-) - trans-2- (2-Chloro-4-nitrophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one hydrochloride

(This compound is the (-) - trans enantiomer of the compound illustrated in Example 50. The (-) - trans Enantiomer was obtained to compare its activity with its corresponding (+) - trans enantiomer.).

(BUT). (-) - trans-2-chloro-4-nitrobenzoic acid 2- (2-acetoxymethyl-1-methylpyrrolidin-3-yl) -6-acetyl-3,5-dimethoxyphenyl ester

2-Chloro-4-nitrobenzoic acid (3.55 g, 17 mmol) and thionyl chloride (2 ml) were heated together at 85 ° C for 3 hours. The excess thionyl chloride was evaporated under reduced pressure, and the residue was dried in vacuo and dissolved in THF (50 ml). It was added dropwise to a solution of the compound of Example (96- (B)) (4 g, 11 mmol) and triethylamine (15 ml, 108 mmol) in THF (100 ml), which was maintained at 15-20 ° C. The reaction mixture was warmed to room temperature and stirred for 12 hours. At the end of 12 hours, the reaction mixture was concentrated to remove THF, acidified with dilute HCl, basified with 10% NaHCO ₃ solution to pH 10, extracted with EtOAc (3 × 100 ml), washed with water (50 ml), brine (50 ml) and the organic layer dried over anhydrous Na ₂ SO ₄ . The organic layer was concentrated under reduced pressure to obtain the title compound as an oil, which was used as such in the next reaction without purification.

Yield: 1.62 g (25%); ¹ H NMR (CDCl ₃ , 300 MHz): δ 8.35 (d, 1H), 8.25 (m, 2H), 6.45 (s, 1H), 4.10 (m, 1H), 3, 94 (s, 6H), 3.59 (m, 1H), 3.17 (m, 1H), 2.70 (m, 1H), 2.49 (s, 3H), 2.46 (m, 2H ), 2.35 (s, 3H), 2.17 (m, 1H), 1.95 (m, 1H), 1.83 (s, 3H); MS (ES +): m / z 535 (M + 1).

(AT). (-) - trans-2- (2-Chloro-4-nitrophenyl) -8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -5,7-dimethoxychromen-4-one

Hexamethyldisilazane (1.25 ml, 5.8 mmol) was added dropwise to a solution of n-BuLi (15% in hexane, 2.54 ml, 5.8 mmol) in THF (25 ml), which was maintained at 0 ° C under a nitrogen atmosphere. ) and mixed for 15 minutes. To this was added dropwise a solution of (-) - trans-2-chloro-4-nitrobenzoic acid 2- (2-acetoxymethyl-1-methylpyrrolidin-3-yl) -6-acetyl-3,5-dimethoxyphenyl ester (1, 6 g, 3 mmol) in THF (25 ml), maintaining the temperature at 0 ° C. After addition, the reaction was warmed to room temperature and stirred for 2.5 hours. The reaction mixture was acidified with dilute HCl, made basic with 10% NaHCO ₃ solution to pH 8-9. The aqueous layer was extracted with chloroform (3 × 50 ml). The organic layer was washed with water (25 ml), brine (25 ml), dried over anhydrous Na ₂ SO ₄ . The organic layer was concentrated under reduced pressure and under vacuum to obtain 3- {3- [3- (2-chloro-4-nitrophenyl) -3-oxopropionyl] -2-hydroxy-4,6-dimethoxyphenyl} -1-methylpyrrolidine acetic acid -2-yl-methyl ester as a yellow solid (1.6 g). This ester was dissolved in conc. HCl (16 ml) and stirred at room temperature for 3 hours to perform cyclization. At the end of 3 hours, the reaction mixture was basified with solid NaHCO ₃ to pH 8-9. The aqueous layer was extracted with chloroform (3 × 50 ml) and washed with water (25 ml) and brine (25 ml). The organic layer was dried over anhydrous Na ₂ SO ₄ ; concentrated under reduced pressure and dried under vacuum. The residue was purified by column chromatography with 3% methanol in chloroform and 0.1% ammonia as an eluent to obtain the title compound as a yellow solid.

Yield: 0.71 g (50%); IR (KBr): 3447, 1648, 1600, 1570 cm ^-1 ; ¹ H NMR (CDCl ₃ ): δ 8.37 (d, 1H), 8.26 (dd, 1H), 8.09 (d, 1H), 6.67 (s, 1H), 6.46 (s , 1H), 4.18 (m, 1H), 4.01 (s, 3H), 3.98 (s, 3H), 3.67 (m, 1H), 3.32 (m, 1H), 3 19 (m, 1H); 2.62 (m, 2H); 2.36 (s, 3H); 2.04 (m, 2H); MS (ES +): m / z 475 (M + 1).

(C) (-) - trans-2- (2-Chloro-4-nitrophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one

A mixture of (-) - trans-2- (2-chloro-4-nitrophenyl) -8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -5,7-dimethoxychromen-4-one (0.4 g, 0.8 mmol), pyridine hydrochloride (0.48 g, 4.1 mmol) and a catalytic amount of quinoline were heated at 180 ° C for a period of 2 hours. The reaction mixture was cooled and diluted with methanol (50 ml) and made basic with solid Na ₂ CO ₃ to pH 10. The reaction mixture was filtered and washed with methanol. The organic layer was concentrated and the residue was purified by column chromatography using 0.1% ammonia and 3% methanol in chloroform as an eluent to obtain the title compound as a yellow solid.

Yield: 0.24 g (65%); MS (ES +): m / z 447 (M + 1)

(D) (-) - trans-2- (2-Chloro-4-nitrophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one hydrochloride

(-) - trans-2- (2-Chloro-4-nitrophenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one (0.2 g 0.4 mmol) was suspended in methanol (10 ml), treated with ethereal HCl and the organic solvent was evaporated to obtain the title salt.

Yield: 0.18 g (85%); ¹ H NMR (CD ₃ OD, 300 MHz): δ 8.51 (d, 1H), 8, (dd, 1H), 8.09 (d, 1H), 6.69 (s, 1H), 6, 41 (s, 1H), 4.25 (m, 1H), 3.91 (m, 2H), 3.73 (m, 1H), 3.70 (m, 1H), 3.63 (m, 1H ), 3.00 (s, 3H), 2.55 (m, 1H), 2.32 (m, 1H); MS (ES +): m / z 483 (M + 1).

Example 98: (Comparative example: obtained by the method described in published patent application US No. 2004/0106581)

(-) - trans- [1-Methyl-3- (2,4,6-trimethoxyphenyl) pyrrolidin-2-yl] methanol

The compound of formula 4 (27.3 g, 97.1 mmol) was dissolved in methanol (100 ml) and heated to 70 ° C. To this was added a hot solution of (+) DBTA (36.51 g, 101.9 mmol) and heating was continued for 10 minutes. It was concentrated to obtain a solid (63.81 g), which was crystallized using methanol (45 ml) and isopropanol (319 ml). Filtration and washing with isopropanol, followed by drying, provided the tartrate crystalline salt (13.14 g), [α] _D ²⁵ = + 55.34 ° (c = 1.14, methanol). This product was recrystallized using methanol (10 ml) and isopropanol (40 ml). It was isolated as described above, yield: 9.04 g, [α] _D ²⁵ = + 49.67 ° (c = 1.248, methanol). The free base was obtained from this product as follows.

Salt (9 g) was suspended in ethyl acetate (100 ml). To this suspension was added a 5% aqueous solution of NaHCO ₃ (100 ml) and the mixture was stirred for 30 minutes. The organic portion was separated and the aqueous portion was further extracted using ethyl acetate (2 × 50 ml). The organic parts were combined and concentrated to give the compound (-) - trans- [1-Methyl-3- (2,4,6-trimethoxyphenyl) pyrrolidin-2-yl] methanol.

Yield: 3.6 g (26.3%).

[α] _D ²⁵ = -17.6 ° (c = 1.1, methanol).

¹ H NMR (CDCl ₃ ): δ 6.13 (s, 2H), 3.90 (m, 1H), 3.79 (s, 9H), 3.51 (dd, 1H), 3.38 (d , 1H), 3.13 (m, 1H), 2.69 (m, 1H), 2.47 (m, 1H), 2.34 (s, 3H), 2.00 (m, 1H), 1 93 (m, 1H).

MS (ES +): m / z 282 (M + 1).

This compound was subjected to chiral HPLC. Chiral HPLC was performed using a Chiralcel OD-H column (250 × 4.6 mm) and a hexane: ethanol solvent system (92:08) with TFA (0.4%). The results were recorded at 264 nm with a solvent flow rate of 1 ml / min. As depicted in FIG. 2, chiral HPLC shows 88.3% e.i. compounds of (-) - trans- [1-Methyl-3- (2,4,6-trimethoxyphenyl) pyrrolidin-2-yl] methanol.

Biological research:

The effectiveness of the (+) - trans enantiomers of compounds of formula I in inhibiting the activity of cyclin-dependent kinases can be studied by a variety of pharmacological studies well known in the art, such as those described below or, for example, in Losiewics, M.D., et al. Biochem. Biophys. Res. Commun., 1994, 201, 589. The kinases, cyclins, and substrates used in the in vitro kinase study can be proteins isolated from mammalian cells, or alternatively, they can be recombinantly generated proteins. Illustrated pharmacological studies, as described below in this description, were carried out with the compounds of this invention and their salts.

CDK4 / cyclin D1 kinase assay and CDK2 / cyclin E kinase assay

The study measures the phosphorylation of retinoblastoma protein (Rb) CDK4 or CDK2 when activated by cyclin D1 or cyclin E, respectively, by transferring (γ ³² P) - phosphate from γ ³² P-ATP in a study in a 96-well filter plate.

Materials:

CDK4 or CDK2 were co-expressed with cyclin D1 or cyclin E, respectively, by the baculovirus expression system in insect cells. For this, 1 × 10 ⁷ Sf9 cells were co-infected with baculoviruses containing human CDK-4 or 2 and cyclin D1 or E genes, and after 72 hours the cells were lysed in 500 μl of lysis buffer (50 mM HEPES (N-2-hydroxyethylpiperazine-N- 2-ethanesulfonic acid) (pH 7.5), 10 mM MgCl ₂ , 1 mM DTT (dithiothreitol), 5 μg / ml aprotinin, 5 μg / ml leupeptin, 0.1 mm NaF, 0.2 mm phenylmethyl sulfonyl fluoride (PMSF) and sodium orthovanadate. The centrifuged lysate was purified on a GST (glutamine cyclotransferase) -sepharose column. Protein purity was applied using SDS-PAGE followed by Western blots using specific antibodies (Santacruz Biotec, USA) for CDK4 or CDK2.

The GST retinoblastoma (Rb) fusion protein (aa 776-928) was expressed in E. coli bacteria and purified by GSH Sepharose affinity chromatography. GST-Rb bound to these granules served as a substrate in the study.

Reading:

Quantitative analysis was carried out by scintillation determination of ( ³² P) -GST-Rb in 96-well filter plates using a 96-well Top Count scintillation counter (Packard, USA).

Methodology:

The study of the enzyme CDK4 or CDK2 was carried out in a 96-well form using multi-sieve filter plates with an ultrafine filter. All stages of the study were carried out in a tablet with one filter (tablets Unifilter, Packard, USA). The filter wells were pre-wetted with kinase buffer (100 □ L / well) and the solution was then removed using vacuum, with a filter plate on a vacuum manifold, and the vacuum was turned on. 50 μl of GST-Rb bound to GSH-Sepharose granules in kinase buffer (0.5 □ g GST-Rb / 50 μl) was added to each well and a vacuum was applied to remove the buffer. An additional 25 μl of the reaction mixture containing ATP (cold + hot) and phosphatase inhibitors diluted in kinase buffer was added to each well, followed by the addition of the test compound (4 × final concentration in kinase buffer) or kinase buffer (control) in an additional 25 μl volume. A final 50 μl (100 ng) of human CDK-4 / D1 or CDK-2 / E enzyme in kinase buffer was added to each well to initiate the reaction. The reaction was incubated for 30 minutes at 30 ° C. After completion of the reaction, a vacuum was applied and the plate was washed with wash buffer (TNEN buffer) three times. The filter plate was air dried and placed in a multi-sieve connection plate. 30 μl Packard Microscint-O cocktail was added to each well and the plate was coated with Top-Seal A. The plate was read in a Packard Top Count Scintillation scintillation counter for 10 minutes. Flavopiridol was used as a standard inhibitor in all experiments.

The concentration of the compound at which 50% of the phosphokinase activity of CDK4-cyclin D1 and CDK2-cyclin E was inhibited (IC ₅₀ ) was calculated for representative compounds and their pharmaceutically acceptable salts described in the examples. The results are shown in table 1.

Table 1: Compound (s) IC ₅₀ (µM) CDK4-CYCLINE D1 CDK2-CYCLIN E The compound of formula 10 0,065 2,8 The compound of formula 96- (F) NA 21 The compound of formula 50 0,045 1.7 The compound of formula 97- (D) one > 10 Flavopiridol 60 360

In vitro cell proliferation and cytotoxicity assays:

Used exponentially growing cultures of ten lines of human cancer cells (Calu1 lung, A-549 lung, NT-29 colon, HL-60 promyelocytic leukemia, PC-3 prostate, H-460 lung, MDA-MB-231 breast, MCF- 7 breast, HeLa of the cervix, Colo-205 colon, H9 lymphoma (T Cells), U-937 histiocytic lymphoma (monocytes) and CaCO-2 Colon) obtained from NCCS, Pune, India. In vitro cell proliferation (NCI, USA protocol) and cytotoxicity studies were performed using standard techniques, namely ³ H-thymidine uptake and cytotoxicity using MTT (tetrazolium (3- [4,5-dimethylthiazol-2-yl] -2 , 5-diphenyltetrazolium bromide), respectively (For ³ H-thymidine uptake: Cell Biology, A Laboratory Handbook, 1998, Vol 1 Ed Julio E. Celis, and for MTT studies: Promega Protocol, USA, 2000). In Absorption Study ³ H-thymidine cells were harvested after 72 hours on GF / B unifilter plates (Packard, USA) using a Packard Filtermate Universal cell collector and plates were read at 96- unochnom liquid scintillation counter Packard TopCount. The concentration of compound at which 50% of proliferative activity was inhibited, (IC _50), and the degree of toxicity of compound were calculated for representative compounds and their pharmaceutically acceptable salts described in the examples. The results are shown in Table 2 below.

Table 2: Compound (s) Thymidine Test (IC ₅₀ μM) MTT cytotoxicity (IC ₅₀ μM) Calu1 (Light) A-549 (Light) NT-29 (Large Intestine) Calu1 (Light) A-549 (Light) NT-29 (Large Intestine) The compound of formula 10 <0.3 0.18 0.68 0.88 0.7 0.58 The compound of formula 96- (F) * NA 10 25 m 16.3 16 17.9 The compound of formula 50 <0.3 1,2 1,5 1.4 1,2 1,1 The compound of formula 97- (D) Nd * NA 10 Nd * NA * NA * no statically significant inhibition was observed NA - Inactive ND - Not finished

The results show that the (+) trans-enantiomer of the compounds of formula (I) is a stronger inhibitor than its (-) trans-enantiomer. Thus, the administration of the (+) - trans-enantiomer of the compounds of formula (I), mainly the free (-) - trans-enantiomer, reduces the required dose of the drug.

Claims (9)

1. An enantiomerically pure compound represented by the following formula (I), or a pharmaceutically acceptable salt thereof,

where R ₁ represents phenyl, which is substituted by 2 different substituents selected from: halogen, fluoromethyl, difluoromethyl and trifluoromethyl, provided that if one of the substituents on the phenyl group is halogen, the other substituent is not halogen;
R ₂ and R _{3 are} each independently selected from: hydroxyl and OR ₈ ; where R ₈ represents a substituted or unsubstituted C ₁ -C ₁₀ alkyl, C ₁ -C ₄ alkanoyl, substituted or unsubstituted aroyl;
R ₄ represents a C ₁ -C ₄ alkylene hydroxyl; and
R ₉ represents hydrogen or C ₁ -C ₄ alkyl. 2. The enantiomerically pure compound of formula I according to claim 1, or a pharmaceutically acceptable salt thereof, selected from
(+) - trans-2- (2-chloro-4-trifluoromethylphenyl) -8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -5,7-dimethoxychromen-4-one;
(+) - trans-2- (2-chloro-4-trifluoromethylphenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one;
(+) - trans-2- (2-chloro-4-trifluoromethylphenyl) -5,7-dihydroxy-8- (2-hydroxymethyl-1-methylpyrrolidin-3-yl) -chromen-4-one hydrochloride. 3. Enantiomerically pure dissolved compound represented by the formula (VIIIA)

where R ₁ represents phenyl substituted with two different substituents selected from halogen, fluoromethyl, difluoromethyl and trifluoromethyl, provided that if one of the substituents on the phenyl group is halogen, the other substituent is not halogen;
R ₂ and R _{3 are} each independently selected from hydroxyl and OR ₈ , where R ₈ is substituted or unsubstituted C ₁ -C ₁₀ alkyl, C ₁ -C ₄ alkanoyl, substituted or unsubstituted aroyl;
R ₄ represents CH ₂ OS (O) CH ₃ ; and
R ₉ represents hydrogen or C ₁ -C ₄ alkyl. 4. Enantiomerically pure dissolved compound represented by the formula (IXA)

where R ₁ represents phenyl substituted with two different substituents selected from halogen, fluoromethyl, difluoromethyl and trifluoromethyl, provided that if one of the substituents on the phenyl group is halogen, the other substituent is not halogen;
R ₂ and R _{3 are} each independently selected from hydroxyl and OR ₈ , where R ₈ is substituted or unsubstituted C ₁ -C ₁₀ alkyl, C ₁ -C ₄ alkanoyl, substituted or unsubstituted aroyl;
R ₄ represents CH ₂ OS (O) CH ₃ ; and
R ₉ represents hydrogen or C ₁ -C ₄ alkyl. 5. Enantiomerically pure dissolved compound represented by the formula (XA)

where R ₁ represents phenyl substituted with two different substituents selected from halogen, fluoromethyl, difluoromethyl and trifluoromethyl, provided that if one of the substituents on the phenyl group is halogen, the other substituent is not halogen;
R ₂ and R _{3 are} each independently selected from hydroxyl and OR ₈ , R ₈ is substituted or unsubstituted C ₁ -C ₁₀ alkyl, C ₁ -C ₄ alkanoyl, substituted or unsubstituted aroyl;
R ₄ represents CH ₂ OH; and
R ₉ represents hydrogen or C ₁ -C ₄ alkyl. 6. A method of obtaining an enantiomerically pure compound of the formula (VIA)

comprising the step of reacting the racemate of a compound of formula (VIA) with (-) - dibenzoyl tartaric acid ((-) - DBTA) as a chiral additive in methanol as a solvent to obtain a mixture of diastereomeric salts (+) - and (- ) -trans enantiomers of the compounds of formula (VIA), separating the corresponding diastereomeric salts and treating the diastereomeric salt of the (-) - trans enantiomer with a base. 7. A method of obtaining enantiomerically pure (+) - trans enantiomer of a compound of formula (I) according to claim 1 or claim 2, comprising the steps of
(a) treating the enantiomerically pure dissolved (-) - trans enantiomer of the compound represented by formula (VIA)

where R ₂ , R ₃ R ₄ and R ₉ are as in claim 4, with acetic anhydride in the presence of a catalyst of a Lewis acid of boron trifluoride, zinc chloride, aluminum chloride and titanium chloride selected from diethyl ether to obtain a dissolved acetylated compound of the formula (VIIA )

where R ₂ , R ₃ and R ₉ are as defined, and R ₄ represents CH ₂ OC (O) CH ₃ ;
(b) reacting the dissolved acetylated compound of formula (VIIA) of step (a) with an acid of formula R ₁ COOH, or an acid chloride of formula R ₁ COCl,
or an acid anhydride of the formula (R ₁ CO) ₂ O, or an ester of the formula R ₁ COOCH ₃ , where R _{1 is} defined above, in the presence of a base and a solvent to obtain a compound of formula (VIIIA)

where R ₁ , R ₂ , R ₃ and R ₉ are as defined above, and R ₄ represents CH ₂ OC (O) CH ₃ ;
(c) treating the dissolved compound of formula (VIIIA) with a base selected from lithium hexamethyldisilazide, sodium hexamethyldisilazide, potassium hexamethyldisilazide, sodium hydride and potassium hydride to obtain the corresponding dissolved β-diketone compound of formula (IXA)

where R ₁ , R ₂ , R ₃ and R ₉ are as defined above, and R ₄ represents CH ₂ OC (O) CH ₃ ;
(d) treating the compound of β-diketone of formula (IXA) with acid to obtain the corresponding dissolved cyclic compound of formula (XA)

where R ₁ , R ₂ , R ₃ and R ₉ are as defined above, R ₄ represents CH ₂ OH;
(e) treating the dissolved compound of formula (XA) with a dealkylating agent selected from pyridine hydrochloride, boron tribromide, boron trifluoride etherate and aluminum trichloride for the dealkylation action at a temperature in the range of 120-180 ° C. to obtain enantiomerically pure (+) trans enantiomer compounds of formula (I). 8. A pharmaceutical composition for treating cancer comprising a therapeutically effective amount of an enantiomerically pure compound of formula (I) according to claim 1 or 2, or a pharmacologically acceptable salt thereof, and a pharmaceutically acceptable carrier and, optionally, an additive. 9. The use of the enantiomerically pure compound of formula (I) according to claim 1 or 2, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for treating cancer in a mammal.

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