WO2008007210A2 - Nitrogenous n-substituted 4-spiro-heterocyclic 2, 2 -dimethylchromane derivatives - Google Patents

Abstract

Compounds having the following general formula : wherein R0, R1, R2, A and Y are as defined in the specification. The compounds can be used for therapeutic or prophylactic treatments of biological tissues of anatomical parts, such as heart, the pancreatic beta cells, the smooth muscle, the kidney and the central nervous system, involved with ischemic episodes.

Description

TITLE

NITROGENOUS N-SUBSTITUTED 4-SPIRO-HETEROCYCLIC 2,2-DIMETHYLCHROMANE DERIVATIVES

DESCRIPTION Ambito of the invention

The present invention relates to a class of compounds that can be used for therapeutic or prophylactic treatments of biological tissues of anatomical parts, such as heart, the pancreatic beta cells, the smooth muscle, the kidney and the central nervous system, involved with ischemic episodes. Description of the prior art

As known, myocardial ischemia is caused by a substantial reduction of the blood flow and a subsequent insufficient oxygen supply, owing to phenomena such as a constriction of coronary artery vessels, or to the production of clots, or still to the production of atherosclerotic plaques.

The myocardial ischemia determines, in particular, pathological cellular phenomena, which cause tissue damage and, eventually, cell necrosis. In an early stage, when the tissue damage is still reversible, a further and irreversible damage can be prevented with the use of surgical or pharmacologic treatments, for the reinstatement of the blood flow. However, this reinstatement can as such cause a tissue damage, the so-called "reperfusion injury", due to negative effects of oxygen radicalic reagents species and to calcium overload. It is also known that with the expression "ischemic preconditioning" the capacity is commonly described of the myocardium of detecting, after one or more short ischemic episodes, an augmented tolerance with respect to a successive ischemic episode. An augmented tolerance to temporary hypoxia has been observed, in particular, by detecting a minor ventricular disfunction and a rise of the myocardial ischemic threshold in the presence of an equivalent flow reduction. Therefore, researches are being made for a pharmacological induction of this phenomenon.

In particular, it has been widely shown that selective agonists of the δ-opioid receptor are involved in ischemic preconditioning phenomena. This has brought to the formulation of different pharmaceutical compositions having cardioprotective effects mediated by agonists of δ-opioid receptors that induce pharmacologically a ischemic preconditioning. An example of a composition of this type is described in US 6103722 in the name of Schultz and Gross.

However, such compositions have different side effects, in particular they can cause convulsions whenever they pass the blood-brain barrier.

Among the studies for determining possible pharmacologic treatments for reduction of the ischemic myocardial damage and the reperfusion injury, it has been also found that diazoxide, a known activator of ATP and KATP sensitive potassium channels, has this pharmacodynamic profile.

The KATP channels, widely expressed in many organs and tissues, such as heart, the pancreatic beta cells, the smooth muscle, the kidney and the central nervous system, have been considered as an interesting pharmacological target for development of different therapeutic classes, and presently many chemically heterogeneous KATP-activator classes have been described (Mannhold, 2006, Curr. Topics Med. Chem. 6, 1031-1047) .

However, such compositions do not have a significant selectivity and, concerning their use as anti-ischemic drugs, their cardio-protective activity is assocyanided with a wide variety of undesirable side effects, that involve many other anatomical . zones .

Summary of the invention It is therefore according to the present invention to provide a pharmaceutical composition, for therapeutic or prophylactic treatments of biological tissues involved with ischemic episodes, which has not the typical side effects of the cardioprotective drugs of the prior art and that pharmacologicically induce an ischemic preconditioning.

It is another feature of the present invention to provide such a pharmaceutical composition that is capable of considerably reducing the tissue damage caused to a biological tissue by a long obstruction of the blood perfusion.

It is also according to the present invention to provide such a pharmaceutical composition that is much more selective for myocardium than the drugs of prior art, and that in particular has a partial but significant action at vascular level and without or low secondary activity concerning other biologically relevant functions.

These and other features are accomplished with one exemplary compound, according to the invention, whose main feature is to have the following general formula (X) :

wherein

— * represents a chiral center.

— Ro is selected from the group comprised of: a carbonyl or thiocarbonyl group, an alkyl group (methylene, ethylene, or methylcarbonyl or methyl- thiocarbonyl group.

— Y is selected from the group comprised of: a CH₂ group, a C=O group, a C=S group, a C=NH group.

— A is selected from the group comprised of: a - CONH- group, a -COO- group, a -CO- group, an alkyl group (C1-C3) , an alkylcarbonyl group, a carbonyl group, a thiocarbonyl group, an alkylthiocarbonyl group, a sulphonic group, an alkylsulphonic group.

— Ri is selected from the group comprised of: a hydrogen atom, an alkyl group, (methyl, ethyl, propyl, isopropyl, butyl, iso-butyl or tert-butyl, an alkoxy group (methoxy, ethoxy, n-propyloxy, iso-propyloxy) , an halide atom (F, Cl, Br, I), a trifluoromethyl group, a cyanide group, a nitro group, a hydroxy group, an amine group, an alkylamine group, an alkylamide group (acetamide, trifluoroacetamide, propionamide) or alkyl- sulphonamide (methane-sulphonamide, ethane- sulphonamide) . — R₂ is selected from the group comprised of: a hydrogen atom, an alkyl group with C1-C4 carbon atoms (methyl, ethyl, propyl, isopropyl, butyl, isobutyl or terbutyl) , a carboxyl group, an alkoxy group (methoxy, ethoxy, n-propyloxy, iso- propyloxy) , an halide atom- (F, Cl, Br, I) , a cyanide group, a nitro group, a trifluoromethyl group, a hydroxy group, a thioalkyl group (Cl, C2,

C3) ; an amine group of NR3R4 type where R3 and R4 can be indifferently a hydrogen atom, an alkyl, alkylsulphonic (methanesulphonic, ethanesulphonic) , acyl (acetyl, propionyl) , trifluoroacetyl group.

In particular, R₀ can be a -CH₂CO- group and Y a -

CH₂- group. Alternatively, Ro can be a - (CH₂) ₂- group, and Y a

-CH₂- type group.

In a further exemplary embodiment, Ro can be a C=O group and Y selected from the group comprised of: -

CH₂- group, C=O group, C=NH group. In particular, Y can comprise one of the above described nitrogen substituted groups for a substituting group selected from the group comprised of: an alkyl group, an acyl group, a benzyl group.

More in detail, also the substitute groups can be in turn substituted with groups such as alkyl, amine or alkylamine (C1-C4), amide, N-alkylsulphonamide or N- alkylamide, halide (Cl, Br, F, I), alkyl halide

(trifluoromethyl) , or electron withdrawing groups

(nitro or cyanide) . In particular, the above described compounds can comprise not only a racemic mixture but also single enantiomers .

According to another aspect of the invention a pharmaceutical composition for treatment of biological tissues involved with ischemic episodes comprises a measured amount of at least one compound having general formula (X) as above shown and described.

Advantageously, in addition to at least one of the above described compounds the pharmaceutical composition comprises also pharmacologically acceptable excipients.

In particular, the pharmaceutical composition is adapted to induce a ischemic preconditioning through the activation of KATP channels.

Hereafter examples are given of a possible synthesis process of the compounds according to the present invention. In particular, the compounds having above general formula can be prepared through the following reaction scheme:

[[ IU

stepd

VII VIII

IX X wherein

— Rl can be, similarly to the general formula, a hydrogen atom, an alkyl group (methyl, ethyl, propyl, isopropyl, butyl, isobutyl or tertbutyl, an alkoxy group (methoxy, ethoxy, n-, iso-propyloxy) , an halide atom (F, Cl, Br, I) , a cyanide group, a nitro group, a hydroxy group, an amine group, an alkylamine group, an alkylamide group (acetamide, trifluoroacetamide, propionamide) or alkylsulphonamide (methane- sulphonamide, ethane-sulphonamide) . — R can be a cyanide or a thiocyanide group.

— R₅ and R₆ can be indifferently hydrogen atoms, alkyl groups (C1-C4), alkylsulphonic groups, alkylarylsulphonic, acyl groups and carbamic groups. All these groups can also be substituted with not substituted aromatic groups or suitably substituted aromatic groups for a group R₂ that is the same as above described with reference to the general formula.

— Xi represents a hydrogen atom, a suitable leaving group (trimethylsilyl, mesyl, tosyl) or an alkaline or alkaline earth metal.

— Y can be a CH₂, C=O, C=S, C=NH group.

— B represents a carbonyl group, an alkylcarbonyl group, or an alkyl-thiocarbonyl group; it can, furthermore, be the same as Ro described in the general formula.

— Z and Z' can be indifferently an halide atom (Cl, Br, I) or an alkoxy group (methoxy, ethoxy) or suitable leaving groups, among these groups, an alkansulphonyloxy or arylsulphonyloxy group.

— Ro represents one of the groups already defined in the general formula.

In short, the reaction scheme given above begins with the reaction of compound I with acetone. This causes the production of a suitably substituted chromanne derivative II, which owing to nucleophilic addition on the carbonyl group provides a derivative III that is then reduced to derivative IV. This is then subject to acylation, or to alkylation, with a suitable V agent in the presence of a base. The compound VI thus obtained is subject to a reaction of cyclization and following reduction to compound VII.

Hereinafter the different steps are described in the detail, i.e. steps a-h, of the reaction scheme above indicated. Step a

In particular, the reaction of compound I with acetone that causes the production of compound II can be done in a wide temperature range, since this process parameter is not critical for the reaction. In general, the reaction can be carried out at a temperature comprised in a range between 0 and 200⁰C. The reaction, furthermore, is preferably carried out in the presence of solvents such as nitriles, for example acetonitrile; aromatic hydrocarbons, for example toluene, benzene or xylene; amides such as dimethylacetamide, ethers such as tetrahydrofuran, dioxane or ethyl ether, ketones such as acetone.

The reaction is carried out in the presence of a base such as an alkaline hydroxide, such as sodium hydroxide or potassium hydroxide; an amide, such as sodium amide or potassium amide; an alkoxide such as sodium methoxide, sodium ethoxide or potassium t- butoxide; an organic base such as triethylamine, the pyrrolidine, the N-methylpyrrolidine, the pyridine.

The temperature of reaction, the nature of the solvents and reagents used affect the reaction time that can be adjusted from 30 minutes to 24 hours.

At the end of the reaction the desired compound of formula II can be extracted from the reaction mixture by conventional methods. For example, an extraction procedure is shown by filtering any insoluble material (if present) and evaporating the solvent at low pressure to obtain the desired product. An alternative embodiment provides the evaporation of the solvent at low pressure, and the residue material is treated with a water immiscible organic solvent (For example ethyl acetate, dichloromethane) and washed with water. The organic phases are gathered are dried with a dehydrating agent such as sodium sulphate and finally the solvent has evaporated. The residue, if necessary, can be further purified by conventional methods such as crystallization or chromatographic techniques. Step b

The carbonyl group of compound II is then subject to a reaction of nucleophilic addition to obtain compound III. This reaction is preferably carried out in the presence of a solvent. Examples of solvents that can be used are the aliphatic halogenated hydrocarbons such as dichloromethane, chloroform, ethers, such as ethyl ether, tetrahydrofuran or dioxane, aromatic hydrocarbons such as toluene, benzene or xylene; aliphatic hydrocarbons such as hexane or the cyclohexane. a mixture can also be used of one or more than the above cited solvents.

The reaction is carried out in the presence of a catalyst, preferably a Lewis acid such as zinc iodide, the aluminium trichloride or lithium perchlorate. Even this reaction can be carried out in a wide temperature range. In particular, the reaction can be carried out at a temperature set between 20 and 80⁰C. The time required for the reaction depends on the nature of the reagents and of the solvents used. Normally, in the conditions above described, a time set between 30 minutes and 48 hours is sufficient.

At the end of the reaction the desired compound of formula III can be extracted from the reaction mixture by conventional methods. For example, an extraction procedure is shown by filtering insoluble materials

(if present) and then evaporating the solvent at low pressure to provide the desired product. An alternative embodiment provides the evaporation of the solvent at low pressure, and the residual matter is treated with a water immiscible organic solvent (For example ethyl acetate, dichloromethane) and washed with water. The gathered organic phases are dried with a dehydrating agent such as sodium sulphate and finally the solvent has evaporated. The residue, if necessary, can be further purified by conventional methods such as crystallization or chromatographic techniques. Step c

The reaction indicated in this step can be a hydrolysis reaction, or alternatively, a reduction reaction. In the first case, the hydrolysis reaction can be carried out in the presence of solvents for example alkohols such as methanol, ethanol, isopropanol; ethers such as tetrahydrofuran or dioxane, nitriles, such as acetonitrile . The reaction is carried out in the presence of protic acids such as hydrochloric acid, acetic acid or Lewis acid such as boron trifluoride. The reaction can be carried out within a temperature range between 0 to 100⁰C. in the latter case, the reduction reaction can be made by means of catalytic hydrogenation, carried out at a pressure set between the atmospheric pressure and 10 atmospheres, or using reducing reagents such as hydrides (sodium borohydride, lithium aluminium hydride, diborane) . The reaction is preferably carried out in the presence of a solvent, for example alkohols, such as methanol, ethanol, or isopropanol; ethers, such as ethyl ether, tetrahydrofuran or dioxane; aromatic hydrocarbons, such as toluene, benzene or xylene; aliphatic hydrocarbons, such as hexane or cyclohexane; esters, such as ethyl acetate and fatty acids such as acetic acid. A mixture can also be used of one or more than the above cited solvents. The reaction can be carried out in a wide temperature range. Generally, it is much more preferable to carry out the reaction in conditions of temperature set between 20 and 80⁰C, for example at room temperature or at 40°, 60⁰C. The time required for the reaction depends on the nature of the reagents and solvents used. Normally in the conditions above described, a time set between 30 minutes and 48 hours, for example 1, 3, 5, 10, 15, 20 or 30 hours is sufficient .

At the end of the reaction the desired compound of formula IV can be extracted from the reaction mixture by conventional methods. For example, an extraction procedure is shown by filtering insoluble materials

(if present) and then evaporating the solvent at low pressure to provide the desired product. An alternative embodiment is the evaporation of the solvent at low pressure, and the residual matter is treated with water and extracted with a water immiscible organic solvent (For example ethyl acetate, dichloromethane) . The gathered organic phases are dried with a dehydrating agent such as sodium sulphate and finally the solvent has evaporated. The residue, if necessary, can be further purified by conventional methods such as crystallization or chromatographic techniques . Step d

A mole of compound IV is caused to react with a base (From 1 to 3 moles) and with a alkylating, or acylating V agent (1-3 moles) depending on whether they the desired reaction is a reaction of alkylation, or acylation respectively. The reaction can be carried out in solvents as dimethylformamide, dimethylacetamide, dimethylsulphoxide, acetonitryl, acetone, ethylmethylketone, and the like. The bases that can be used are sodium hydride, potassium t- butoxide, potassium carbonate, sodium carbonate and the like. Examples of alkylating agents can be alkyl halides (chlorides, bromides, iodides) . The conditions of reaction can be adjusted and depend in general by the combination of the agent and of the base used. The reaction is carried out at a temperature that varies between 0-100⁰C, for example 5°, 10°, 20°, 40°, 60°, 80⁰C and depends on the nature of the reagents, of the solvent and of the base used. The time of reaction is influenced by the type of reagents and solvent used and can be adjusted between 1-48 hours.

At the end of the reaction the desired compound of formula VI can be extracted from the reaction mixture by conventional methods. For example, an extraction procedure is shown by filtering insoluble materials

(if present) and then evaporating the solvent at low pressure to provide the desired product. An alternative embodiment is the evaporation of the solvent at low pressure, and the residual matter is treated with water and extracted with a water immiscible organic solvent (For example ethyl acetate, dichloromethane) . The gathered organic phases are dried with a dehydrating agent such as sodium sulphate and finally the solvent has evaporated. The residue, if necessary, can be further purified by conventional methods such as crystallization or chromatographic techniques . Step e The reaction described in this passage provides that the intermediate product of reaction VI is subject to a reaction of cyclization, or alternatively, to a reaction of nucleophilic substitution. In the first case, the reaction of cyclization is normally carried out in the presence of inert solvents and in the presence of a base. The base may be of many- kinds; the more commonly used are carbonates of alkaline metals, such as sodium carbonate, potassium carbonate; hydrogencarbonates of alkaline metals, such as hydrogen sodium carbonate or hydrogen potassium carbonate; fluorides of alkaline metals, such as sodium fluoride and potassium fluoride; hydrides such as sodium hydride, potassium hydride or lithium hydride; alkoxides as sodium methoxide, sodium ethoxide, potassium t-butoxide, lithium ethoxide; organic amines such as pyridine, picoline, triethylamine, N-methylmorpholine, or 4- dimethylaminopyridine . The reaction is preferably carried out in the presence of a solvent. Examples of solvents used in this reaction are hydrocarbons, such as hexane, toluene or benzene; halogenated hydrocarbons, preferably aliphatic, such as methylene chloride, chloroform, 1, 2-dichloroethane; ethers, such as diethyl ether, tetrahydrofuran, dioxane; ketones, such as acetone, methylethylketone; nitriles, such as acetonitrile; amides, such as dimethylformamide, dimethylacetamide, and sulphoxydes, such as dimethylsulphoxide. Only one of the above cited solvents or a mixture thereof can be used. The reaction is carried out in a wide temperature range set between 0°C and 100⁰C. The time of the reaction can be adjusted and depends on different factors among which the temperature of reaction, the nature of the reagents and of the solvents used. The time of reaction in the conditions above cited can be adjusted in a range set between 30 minutes and the 48 hours.

If an alkyl halide VI, instead, is subject to a reaction of nucleophilic substitution the conditions are similar to the above described for step d and, in particular, the reaction can be carried out using the same reagents and the same conditions of reaction.

Step f The reaction indicated in this step is a reduction reaction that can be carried out with reducing agents or by means of catalytic hydrogenation. The reaction can be carried out using the same reagents and the same conditions of reaction described with reference to the reduction reaction of the step c.

Step g

In step g, a compound having formula X has been obtained for the reaction of compound VIII with a compound of structure IX. This reaction is essentially similar to that described by step d and can be carried out using the same types of reagents and the same conditions of reaction.

Step h

The compound of formula X is transformed in compound XI by a reduction reaction and following alkylation and/or acylation. Alternatively, to the reduction reaction a reaction of nucleophilic substitution can be provided.

In case of a reduction reaction, the same steps are followed as described for step b and for step d. In particular, the same types of reagents and the same conditions of reaction can be used.

In the case, instead, of a nucleophilic substitution reaction, it is carried out in a way similar to that described for step d. The reaction is preferably carried out in the presence of a solvent, not exist particular restrictions concerning the type of solvent used referring to the insolubility of the reagents involved in the reaction. Examples of solvents used are hydrocarbons, such as hexane, benzene or toluene; halogenated hydrocarbons, such as methylene chloride, chloroform, 1, 2-dichloroethane; ethers, such as ethyl ether, tetrahydrofuran, dioxane; ketones, such as acetone, ethylmethylketone; nitriles, such as acetonitrile; amides, such as dimethylacetamide, dimethylformamide, N-methyl-2- pyrrolidone; sulphoxydes, such as dimethylsulphoxide; and water. Only one of the above cited solvents or a mixture thereof can be used. The reaction is carried out in a wide temperature range set between -10⁰C and 100⁰C. The time of the reaction can be adjusted and depends on different factors among which the temperature of reaction, the nature of the reagents and of the solvents used. The time of reaction in the conditions above cited can be adjusted in a range set between 30 minutes and 48 hours.

At the end of the reaction, the desired compound can be extracted from the reaction mixture by conventional methods among which the evaporation of the solvent, or the extraction the product by means of water immiscible organic solvents. The extracted products are then dried with dehydrating agents such as magnesium sulphate or sodium sulphate and evaporated. If necessary, the product can be purified by conventional methods such as crystallization, precipitation or chromatographic techniques. In table 1 some examples of compound are given, according to the invention. For each compound the following are shown: reference code and respective groups R₀, Y, Ri and R₂.

TABLE 1

Other compounds, according to the present invention, are the following: i) 4' - (4-methanesulphonamidobenzyl) -6-bromo-2, 2- dimethyl-2 , 3-dihydro-5' H-spiro [chromen-4 , 2' - [l,4]oxazinane] (F134) . ii) 4' - (4-methanesulphonamidobenzyl) -6-bromo-2, 2, - dimethyl-2, 3-dihydro-5' H-spiro [chromen-4, 2' - [1, 4] oxazinan] -5' -one (F95) . iii) 4' - (4-acetamidobenzyl) -2,2, -dimethy1-2, 3- dihydro-5' H-spiro [chroitιen-4 , 2' - [1, 4] oxazinan] - 5' -one (F163) .

Hereinafter some examples are given of methods of Synthesis of above described compounds and of the intermediates necessary to prepare the same according to the reaction scheme above shown.

EXAMPLE 1: Synthesis of 2 ,2-dimethyl-2 , 3-dihydro- 4H-chromen-4-one (intermediate II)

To a solution of 2-hydroxy acetophenone (1Og, 73.4 mmoles) substituted for toluene (30 ml), acetone (8.11 ml, 0.11 mol) and pyrrolidine (1.84 ml, 23 mmoles) have been added. The solution has been stirred at AT for Ih and then to a reflux for 24 h. After this period, the solvent has been removed at low pressure and the residual matter treated with ethyl acetate. The organic phase has been subject to washings with HCl 6N, NaOH 2N and water. The organic phase, dried, filtered and evaporated to p.r., has provided an oil that has not been subject to further purifications. Yield: 60%

¹H-NMR (CDCl₃) δ: 1.46 (s, 6H); 2.72 (s, 2H); 6.95 (m, 2H); 7.46 (m, IH); 7.85 (m, IH). MS (ffi/z) : 177 (M⁺, 100%)

Elementary analysis:

EXAMPLE 2 : Synthesis of 2 ,2-dimethyl-6-bromo-2 , 3- dihydro-4h-chromen-4-one (intermediate ii) The product has been synthesized as described in the example 1 and has been used in the next reaction without being subject to further purifications.

Yield: 81%

¹H-NMR (CDCl₃) δ: 1.45 (s, 6H); 2.71 (s, 2H); 6.82 (d, IH, J=8.8 Hz); 7.53 (dd, IH, J= 8.8, 2.5Hz); 7.95 (d, IH, J = 2.5 Hz) .

EXAMPLE 3: Synthesis of 4- (trimethylsyliloxy) -2 ,2- dimethylchroman-4-carbonitrile (compound III)

To a solution of 2, 2-dimethyl-2, 3-dihydro-4H- chromen-4-one (4 g, 22 mmoles) in CH₂Cl₂ trimethylsylilcyanide (4.4 ml, 33 mmoles) and ZnI₂ (1.05 g, 3 mmoles) have been added. The mixture has been stirred at AT for 4 h. The solution is then diluted with CH₂Cl₂ and washed with H₂O. The organic phase is dried and evaporated at low pressure to provide a dark raw that corresponds to th₎e _LT desired product .

Yield: (βg, 98%);

¹H-NMR (CDCl₃) δ: 0.27 (s, 9H); 1.44 (s, 3H); 1.48 (s, 3H); 2.35 (d, IH, J=14 Hz); 2.47 (d, IH, J=14 Hz); 6.82-6.86 (m, IH); 6.97-7.05 (m, IH); 7.24-7.32 (m, IH) ; 7.54-7.58 (m, IH) .

Elementary analysis:

Ci₅H_2INO₂Si C H N

Calc% 65 . 42 7 . 69 09

Found% 65 . 15 7 . 71 4 . 78

EXAMPLE 4 : Synthesis of 4- (aminomethyl) -6-bromo- 2 ,2-dimethylchroman-4-ol (compound iv)

In a solution of LiAlH₄ (IM in THF, 31 mmoles) at O⁰C a solution is dropped of 4-hydroxy-6-bromo-2, 2- dimethylchroman-4-carbonitrile (4.23g, 15 mmoles) dissolved in a minimum amount of THF. The solution has been stirred at 0⁰C for 2 h.

After this period to the reaction mixture H₂O and NaOH have been added slowly. After filtering the mixture on a septum, the solution has evaporated at low pressure; the raw oil obtained corresponds essentially to the desired product and is used without further purifications.

Yield: (75%);

¹H-NMR (CDCl₃) δ: 1.35 (s, 3H); 1.42 (s, 3H); 1.98 (s, 2H); 2.78 (d, IH, J=12.8 Hz); 2.96 (d, IH, J=12.8 Hz); 6.71 (d, IH, J=8.8 Hz); 7.26 (dd, IH, J=8.8 Hz, J=2.4 Hz); 7.53 (d, IH, J=2.4 Hz).

MS (m/z) : 286 (M⁺, 16%); 255 (M⁺- 30, 100%).

Elementary analysis

Ci₂Hi₆NO₂Br C H N

Calc% 50 . 37 5 . 64 4 . 89

Found% 50 . 62 5 . 90 4 . 68

EXAMPLE 5: Synthesis of N- [( 4-hydroxy-2 ,2- dimethy1-3,4-dihydro-2H-chromen-4-y1)methy1] -2- chloroacetamide (compound VI)

To a solution of 4- (aminomethyl) -2, 2- dimethylchroman-4-ol (1.64 g, 8 mmoles) in CH₂Cl₂ (24 ml), NaOH (0.38 g, 9.50 mmoles) and H₂O (17 ml) have been added: the mixture has been brought to 0⁰C and chlorideacetylchloride (1.06 ml, 13 mmoles) has been added drop by drop. The mixture has been stirred at AT for about 2 h. The two phases are separated and the organic phase has been washed many times with HCl IN. The gathered organic phases are dried and evaporated; a raw is obtained that corresponds to the desired product.

Yield: 89%

¹H-NMR (CDCl₃) δ: 1.36 (s, 3H); 1.42 (s, 3H); 2.02

(s, 2H); 3.56 (dd, IH, J-13.7 Hz, J=7.2 Hz); 3.73 (dd, IH, J=13.7 Hz, J=5 Hz); 4.08 (s, 2H); 6.82-6.87 (m, IH); 6.92-7.0 (m, IH); 7.18-7.25 (m, IH); 7.41-7.46 (m, IH) .

Elementary analysis

C₁₄Hi₈NO₃Cl C H N

Calc% 59. 26 6. 39 4. 94

Found% 59. 52 6. 18 5. 10

EXAMPLE 6: Synthesis of 2 , 2-dimethyl-2 , 3-dihydro- 5 ' H-spiro [chromen-4 , 2 ' - [1 , 4] oxazinan] -5 ' -one (compound VIII)

To a solution of N- [( 4-hydroxy-2, 2-dimethyl-3, 4- dihydro-2H-chromen-4-yl) methyl] -2-chloroacetamide (2g, 7.05 mmoles) in toluene (100 ml) t-BuOK (4.11 g, 36.69 mrαoles) has been added in portions in the time of 1 h, and the mixture has been stirred at AT for another hour. The solvent has been removed at p.r. and the residual matter treated with ethyl acetate. The organic phase has been washed with H₂O, dried and evaporated, a yellow solid is obtained that has been purified by grinding with ethyl ether.

Yield: 59%

¹H-NMR (CDCl₃) δ: 1.40 (s,3H); 1.42 (s, 3H); 2.04 (d, IH, J=14.6 Hz); 2.42 (d, IH, J=14.6 Hz); 3.25 (dd, IH, J=12.4 Hz, J=4.3 Hz); 3.93 (d, IH, J=12.4 Hz); 4.22 (d, IH, J=17.5 Hz); 4.34 (d, IH, J=17.5 Hz); 6.84-6.92 (m, IH); 6.95-7.0 (m, IH); 7.21-7.29 (m, IH) ; 7.44-7.48 (m, IH) . MS (m/z) : 248 (M⁺, 3%; Elementary analysis:

C₁₄Hi₇NO₃ N

C H

Calc% 68 . 00 6 . 93 5 . 66

Found% 67 . 89 7 . 22 5 . 39

EXAMPLE 7 : Synthesis of 4 ' -benzyl-2,2-dimethyl- 2 , 3-dihydro-5 ' H-spiro [chromen-4 , 2 ' - [1 , 4] oxazinan] -5 ' - one (F77)

To a suspension of NaH (0.27 g, 13 mmoles, dispersed in mineral oil at 60%) in anhydrous DMF (10 ml) in nitrogen environment a 2, 2-dimethyl-2, 3- dihydro-5 ' ff-spiro [chromen-4 , 2 ' - [1, 4] oxazinan] -5 ' -one

(1 g, 4 mmoles) has been added. The mixture has been left at AT for 30 min, then has been brought to 0⁰C and a solution of benzylbromide (0.57 ml, 5 mmoles) in DMF (2 ml) has been added drop by drop. The mixture has been left at AT for 1 h, then diluted with H₂O and AcOEt. The organic phase has been washed many times with ice and with a saturated solution of NaCl, dried and evaporated. The final product has been purified for crystallization in ethyl ether.

Yield: 87% -

¹H-NMR (CDCl₃) δ: 1.09 (s,3H); 1.31 (s, 3H); 1.23 (d, IH, J=14.6 Hz); 1.71 (d, IH, J=14.6 Hz); 3.07(d,lH, 3=12. A Hz); 3.8 (d, IH, J=12.4 Hz); 4.21 (d,lH, J=14.2 Hz); 4.29 (d,lH, J=17.2 Hz); 4.40 (d,lH, J=17.2 Hz); 5.05 (d, IH, J=14.2 Hz); 6.77-6.81 (m, IH); 6.86-6.93 (m, IH); 7.15-7.38 (m, 7H).

Elementary analysis: C₂IH₂₃NO₃ C H N

Calc% 74 . 75 6 . 87 4 . 15

Found% 74 . 56 6 . 80 3 . 88

Example 8 : Synthesis of 4' - (4-nitrobenzyl) -2 ,2- d-^^nethyl-2 , 3-dihydro-5 ' H-spiro [chromen-4 , 2 ' - [l,4]oxazinan]-5' -one (F102)

The compound has been synthesized like in 7 and has been purified by a chromatographic column using a mixture hexane/AcOEt (1:1) as eluent.

Yield: 67%

¹H-NMR (CDCl₃) δ: 1.24 (s, 3H); 1.33 (s, 3H); 1.74 (d, IH, J=14.6 Hz); 2.31 (d, IH, J=14.6 Hz); 3.06 (d, IH, J=12.4 Hz); 3.83 (d, IH, J=12.4 Hz); 4.31 (d, IH, J=Il Hz); 4.40 (d, IH, J=Il Hz); 4.53 (d, IH, J=14.8 Hz); 4.90 (d, IH, J=14.8 Hz); 6.78-6.98 (m, IH); 7.16- 7.24 (in, IH); 7.31-7.36 (m, IH); 7.47 (d, 2H, J=8.8 Hz) ; 8.20 (d, 2H, J=8.8 Hz) .

MS (m/z) : 382 (M⁺, 60%); 177 (100%)

Elementary analysis:

C_2IH₂₂N₂Os C H N

Calc% 65 . 96 5 . 80 7 . 33

Found% 65 . 89 5 . 65 7 . 21

Example 9 : Synthesis of 4'-(4- trifluoromethylbenzyl) -2 , 2-dimethyl-2 , 3-dihydro-5 ' H- spiro [chromen-4 , 2 ' - [1 , 4] oxazinan] -5 ' -one (F161)

The compound has been synthesized like in example 8 and has been purified by grinding with ethyl ether

Yield: 87% 1H-NMR (CDCl₃) δ: 1.18 (s, 3H); 1.33 (s, 3H); 1.71 (d, IH, J=14.6 Hz); 2.27 (d, IH, J=14.6 Hz); 3.05 (d, IH, J=12.4 Hz) ; 3.82 (d, IH, J=12.4 Hz) ; 4.26-4.47 (m, 3H) ; 4.96 (d, IH, J=14.4 Hz) ; 6.79-6.94 (m, 2H) ; 7.20 (dd, IH, J=8.2 Hz, J=I.6 Hz) ; 7.35 (dd, IH, J=7.7 Hz, J=I.6 Hz) ; 7.43 (d, 2H, J=8.2 Hz) ; 7.61 (d, 2H, J=8.2 Hz) .

MS jm/z) : 405 (M⁺, 2%) ; 159 (47%)

Elementary analysis:

C₂₂H₂₂NO₃F₃ C H N

Calc% 65. 18 5. 47 3. 45

Found% 65. 02 5. 50 3. 56

Example 10 : Synthesis of 4 ' -benzyl-6-bromo-2 ,2- dimethyl-2 , 3-dihydro-5 ' H-spiro [chromen-4 , 2 ' - [l,4]oxazinan]-5' -one (F-36)

The compound has been synthesized like in example 9 and has been purified by grinding with ethyl ether.

Yield: 67%

¹H-NMR (CDCl₃) δ: 1.06 (s, 3H); 1.29 (s, 3H); 1.70 (d, IH, J=14.6 Hz); 2.2 (d, IH, J=14.6 Hz); 3.06 (d, IH, J=12.6 Hz); 3.75 (d, IH, J=12.6 Hz); 4.24 (d, IH, J=14.2 Hz); 4.27 (d, IH, J=17.4 Hz); 4.41 (d, IH, J=17.4 Hz); 5.03 (d, IH, J=14.2 Hz); 6.68 (d, IH, J=8.8 Hz); 7.27-7.35 (1, 6H); 7.49 (d, IH, J=2.5 Hz).

MS (m/z) : 417 (M⁺, 22%); 91 (100%)

Elementary analysis:

C₂iH₂₂NO₃Br C H N

Calc% 60. 59 5. 33 3. 36

Found% 60. 27 5. 30 3. 52

Example 11 : Synthesis of 4 ' - (4-nitrobenzyl) -6- bromo-2 , 2-dimethyl-2 , 3-dihydro-5 ' H-spiro [chromen-4 , 2 ' - [l,4]oxazinan]-5'-one (F-106) The compound has been synthesized like in example 10 and has been purified by chromatography and filtering using as eluent a mixture of hexane/acetate

(1:1) •

Yield: 54%

¹H-NMR (CDCl₃) δ: 1.22 (s, 3H); 1.31 (s, 3H); 1.71 (d, IH, J=14.6 Hz); 2.28 (d, IH, J=14.6 Hz); 3.06 (d, IH, J=12.4 Hz); 3.77 (d, IH, J=12.4 Hz); 4.26 (d, IH, J= 17.7Hz); 4.40 (d, IH, J=17.7 Hz); 4.60 (d, IH, J=14.6 Hz); 4.81 (d, IH, J=14.6 Hz); 6.69 (d, IH, J=8.8 Hz); 7.28 (dd, IH, J=8.8 Hz, J=2.4 Hz); 7.42 (d, IH, J=2.4 Hz); 7.48 (d, 2H, J=8.8 Hz); 8.2 (d, 2H, J=8.8 Hz) .

MS (m/z) : 462 (M⁺, 2%); 146 (100%)

Elementary analysis:

C₂iH₂₂NO₃Br C H N

Calc% 54 . 68 4 . 59 6 . 07

Found% 54 . 50 4 . 42 6 . 02

Example 12 : Synthesis of 4'-(4- trifluoromethylbenzyl) -6-bromo-2 , 2-dimethyl-2 , 3- dihydro-5 ' H-spiro [chromen-4 ,2 ' - [1 , 4] oxazinan] -5 ' -one (F165)

The compound has been synthesized like in example 11 and has been purified by grinding with ethyl ether. Yield: 60%

¹H-NMR (CDCl₃) δ: 1.15 (s, 3H); 1.30 (s, 3H); 1.69 (d, IH, J= 14.6 Hz); 2.24 (d, IH, J=14.6 Hz); 3.05 (d,

IH, J=12.4 Hz); 3.77 (d, IH, J=12.4 Hz); 4.28 (d, IH,

J=17.6 Hz); 4.41 (d, IH, J=17.6 Hz); 4.44 (d, IH,

J=14.4 Hz); 4.91 (d, IH, J=14.4 Hz); 6.69 (d, IH,

J=8.6 Hz); 7.27-7.32 (m, IH); 7.41-7. 49 (m, 3H); 7.62 (d, 2H, J=8.0 Hz) . MS (m/z) : 484 (M⁺, 22%); 159 (100%; Elementary analysis:

C₂₂H₂iNO₃F₃Br C H N

Calc% 54 . 56 4 . 37 2 . 89

Found% 54 . 90 4 . 40 57

Example 13 : Synthesis of 4 ' -benzyl-2 ,2-dimethy1- 2 , 3-dihydrospiro [chromen-4 , 2 ' - [1 , 4] oxazinane] (Final, step f) (F-79)

To a IM solution of LiAlH₄ in THF (8.3 mmoles) the compound 4 ' -benzyl-2, 2-dimethyl-2, 3-dihydro-5 ' H- spiro [chromen-4 , 2 ' - [1, 4 ] oxazinan] -5 ' -one (0.7 g, 2.07 mmoles) has been added dissolved in a minimum amount of THF. The mixture has been subject to reflux stirring for 2 hours, then diluted with H₂O (0.15 ml), NaOH (0.15 ml) and H₂O (0.45 ml). The lithi ^ClNum salts have been filtered and washed many times with THF. The organic phase has been evaporated at low pressure for obtaining transparent oil, which has been purified by forming a corresponding clorhydrate. Yield: (450 mg, 60%) . 1H-NMR (CDCl₃) δ: 1.25 (s, 3H); 1.36 (s, 3H); 2.15 (d, IH, J=14.6 Hz); 2.41-2.60 (m, 4H); 2.71-2.77 (m,

IH); 3.37 (d, IH, J=13 Hz); 3.60 (d, IH, J=13 Hz);

3.71-3.80 (m, IH); 3.90-4.02 (m, IH) ; 6.77 (d, IH,

J=8.05 Hz); 6.88-6.96 (m, IH); 7.12-7.20 (m, IH);

^"7.22-7.32 (m, 5H); 7.61 (d, IH, J=7.7 Hz). Elementary analysis:

C21H25NO2 . HCl C H N

Calc% 70 . 08 7 . 28 3 . 89

Found% 70 . 15 7 . 11 4 . 02 Example 14 : Synthesis of 4'-(4- trifluoromethylbenzyl) -2 , 2-dimethyl-2 , 3- dihydrospiro [chromen-4 , 2 ' - [1 , 4] oxazinane] (Fl62)

The compound has been synthesized like in example 13 and has been purified by transformation into clorhydrate and crystallization by MeOH.

Yield: 45%

¹H-NMR (CD₃OD) δ: 1.29 (s, 3H); 1.38 (s, 3H); 2.04 (d, IH, J=15 Hz); 2.72 (d, IH, J=15 Hz); 3.23-3.65 (m, 4H); 3.95-4.18 (m, 2H); 4.46-4.64 (m, 2H); 6.80 (d, IH, J=8.2 Hz); 6.95-7.03 (m, IH); 7.21-7.30 (m, IH); 7.57 (d, IH, J=7.8 Hz); 7.81 (s, 4H).

MS (m/z) : 391 (M⁺,);

Elementary analysis:

Example 15 : Synthesis of 4 ' -benzyl-6-bromo-2 ,2- dimethyl-2 , 3-dihydrospiro [chromen-4 , 2 ' - [1 , 4] oxazinane] ( F-112)

To a solution of 4 ' -benzyl-6-bromo-2, 2-dimethyl- 2, 3-dihydro-5 ' fl-spiro [chromen-4, 2 ' - [1, 4 ] oxazinan] -5 ' - one (0.18 g, 0.43 mmoles) in THF (2 ml) a 2M solution of BH₃ SMe₂ complex in THF (0.15 ml, 1.72 mmoles) has been added.

The mixture has been subject to irradiation with microwaves at 5OW and 70⁰C for 20 min.

After cooling, the solution has been diluted with H₂O and the THF removed at low pressure. The aqueous phase has been acidified, neutralised and finally extracted with EtOAc. The gathered organic phases have been dried and evaporated to provide a dark oil that has been purified by forming a corresponding clorhydrate.

Yield: (87 mg, 46%) .

¹H-NMR (DMSO) δ: 1.22 (s, 3H); 1.25 (s, 3H); 2.40 (d, IH, J=15 Hz); 2.63 (d, IH, J=15Hz); 3.05-3.27 (m, 2H); 3.56-3.66 (m, 2H); 3.88-4.08 (m, 2H); 3.75-3.98 (m, 2H); 4.22-4.46 (m, 2H); 6.74 (d, IH, J=8.7Hz); 7.33-7.51 (m, 4H); 7.67-7.75 (m, 3H).

Elementary analysis:

Example 16 : Synthesis of 4 ' - (4-nitrobenzyl) -6- bromo-2 , 2-dimethyl-2 , 3-dihydrospiro [chromen-4 , 2 ' - [l,4]oxazinane] (F167) The compound has been synthesized like in example 15 and has been purified by transformation into clorhydrate and crystallization by isopropyl ether. Yield: 47% 1H-NMR (CDCl₃) δ: 1.28 (s, 3H); 1.34 (s, 3H); 2.01 (d, IH, J=14.4 Hz); 2.4-2.5 (m, 4H); 2.66-2.73 (m, IH); 3.5 (d, IH, J=13.5 Hz); 3.64 (d, IH, J=13.5 Hz); 3.75-3.98 (m, 2H); 6.66 (d, IH, J=8.6 Hz); 7.23-7.27 (m, IH); 7.52 (d, 2H, J=8.05 Hz); 7.81 (s, IH); 8.18 (d, 2H, J=8.0 Hz) . MS (ffl/z) : 446 (M⁺, 11%); 191 (100%)

Elementary analysis:

EXAMPLE 17 : Synthesis of 4 ' - (4-aminobenzγl) -2 ,2- dimethyl-2 , 3-dihydro-5 ' ff-spiro [chromen-4 ,2 ' - [l,4]oxazinan]-5' -one (step h) (F81) To a solution of 4 ' - (4-nitrobenzyl) -2, 2-dimethyl- 2, 3-dihydro-5 ' H-spiro [chromen-4, 2 ' - [1, 4] oxazinan] -5 ' - one (1.5 g, 4 mmoles) in MeOH (50 ml) coal (0.216 g) and FeCl₃ (a tip of a knife) have been added. The reaction mixture has been brought to 66°C, then hydrated hydrazine (3.22 ml, 66 mmoles) has been added slowly. The mixture has been subject to reflux stirring for one night, then filtered on celite washing many times with MeOH.

The solvent has been concentred, the residual matter treated with chloroform, dried and evaporated, obtaining a yellow solid (1.17 g, 85%). Yield: 85%

¹H-NMR (CDCl₃) δ: 1.14 (s, 3H); 1.31 (s, 3H); 1.71 (d, IH, J=14.6 Hz); 2.22 (d, IH, J=14.6 Hz); 3.06 (d, IH, J=12.6 Hz); 3.74 (d, IH, J=12.6 Hz); 4.11 (d, IH, J=14 Hz); 4.25 (d, IH, J=17.5 Hz); 4.35 (d, IH, J=17.5 Hz); 4.90 (d, IH, J=14 Hz); 6.63 (d, 2H, J=8.4 Hz); 6.77-6.95 (m, 2H); 7.07 (d, 2H, J=8.4 Hz); 7.16-7.21 (m, IH); 7.34-7.38 (m, IH). MS jm/z) : 352 (M⁺, 29%); 106 (100%)

Elementary analysis:

C₂₁H₂₄N₂O₃ C H N

Calc% 71 . 57 6 . 86 7 . 95

Found% 71 . 45 6 . 60 7 . 78

EXAMPLE 18 : Synthesis of 4 ' - (4-aminobenzyl) -6- bromo-2 , 2-dimethyl-2 , 3-dihydro-5 ' h-spiro [chromen-4 ,2 ' - [1,4] oxazinan] -5' -one (scheme d, step h) (F94) The compound has been synthesized like in example 17 and has been purified by grinding in hexane .

Yield: 87%

¹H-NMR (DMSO) δ: 1.10 (s, 3H); 1.23 (s, 3H); 1.64 (d, IH, J=14.6 Hz); 2.30 (d, IH, J=14.6 Hz); 3.03 (d, IH, J=12.6 Hz); 3.78 (d, IH, J=12.8 Hz); 4.08 (d, IH, J=14 Hz); 4.22 (s, 2H); 4.63 (d, IH, J=14 Hz); 6.52 (d, 2H, J=8.2 Hz); 6.80-6.87 (m, IH),-6.95 (d, 2H, J=8 Hz); 7.36 (dd, IH, J=8.4 Hz, J=2,4 Hz); 7.57 (d, IH, J=2.4 Hz) .

Elementary analysis:

C21H23N2O3B3? C H N

Calc% 58 . 48 5 . 37 6 . 4 9

Found% 58 . 27 5 . 50 6 . 60

Example 19: Synthesis of 4 ' - (4-aminobenzyl) -2,2- dimethyl-2 , 3-dihydro-5 ' h-spiro [chromen-4 , 2 ' - [l,4]oxazinane] (scheme b, step f) (F75)

An IM solution in THF of LiAlH₄ (7.95 mraoles) has been added to a solution of 4 '- (4-aminobenzyl) -2, 2- dimethyl-2, 3-dihydro-5 ' H-spiro [chromen-4, 2 ' - [1, 4 ] oxazinan] -5 ' -one (0.70 g, 1.98 mmoles) dissolved in a minimum amount of THF. The mixture has been subject to stirring at 70⁰C for about 2h and then water (0.14 ml), NaOH (0.14 ml) and water (0.43 ml) have been added. The lithium salts have been filtered on a septum and washed (3 times) with THF. The organic phase has been then evaporated to p.r. obtaining a transparent oil that has been purified by transformation into clorhydrate. Yield: 85% 1H-NMR (CDCl₃) δ: 1.26 (s, 3H); 1.35 (s, 3H); 2.12 (d, IH, J=14.6 Hz); 2.43-2.59 (m, IH); 2.70 (d, IH, J=14.6 Hz) ; 3.24 (d, IH, J=12.6 Hz) ; 3.49 (d, IH, J=12.6 Hz) ; 3.60 (s broad, 2H) ; 3.71-3.78 (m, IH) ; 3.87-4.0 (m, IH) ; 6.62 (d, 2H, J=8.2 Hz) ; 6.75-6.79 (m, IH) ; 6.87-6.95 (m, IH) ; 7.08 (d, 2H, J=8.2 Hz) ; 7.15-7.20 (m, IH) ; 7.59-7.62 (m, IH) . MS (m/z) : 338 (M⁺, 4%) ;

Elementary analysis:

C_2IH₂₆N₂O₂ HCl H

C N

Calc% 67 .28 7. 26 1 .47

Found% 67 .52 7. 39 1 .71

Example 20: Synthesis of 4'-(4- methanesulphonamidobenzyl) -2 , 2-dimethyl-2 , 3-dihydro-

5 'H-spiro [chromen-4 ,2 ' - [1, 4] oxazinan] -5' -one (step h) (F82)

To a solution of 4 ' - (4-aminobenzyl) -2, 2-dimethyl- 2, 3-dihydro-5 ' H-spiro [chromen-4, 2 ' - [1, 4] oxazinan] -5' - one (352 g, 1 mmoles) in anhydrous dioxane (10.8 ml) in nitrogen environment pyridine (1.08 ml) has been added, then the solution has been brought to 0⁰C and methanesulphonylchloride (0.1 ml, 1.3 mmoles) has been added slowly. The solution has been reflux stirred for 1 h, then acidified and extracted with EtOAc. The organic phase has been dried and evaporated. The obtained raw product has been purified by grinding with hexane .

Yield: 64% 1H-NMR (CDCl₃) δ: 1.19 (s, 3H); 1.33 (s, 3H); 1.73 (d, IH, J=14.6 Hz); 2.28 (d, IH, J=14.6 Hz); 3.01 (s, 3H); 3.07 (d, IH, J=12.4 Hz); 3.79 (d, IH, J=12.4 Hz); 4.31-4.39 (m, 3H); 4.85 (d, IH, J=14.2 Hz); 6.78-6.94 (m, 2H); 7.17-7.27 (m, 5H); 7.31-7.37 (m, IH). MS (m/z) : 430 (M⁺, 7%); 184 (100%) Elementary analysis:

C₂₂H₂₆N₂O₅S C H N

Calc% 61 . 38 6 . 09 6 . 51

Found% 61 . 52 6 . 00 6 . 42

Example 21: Syn-thesis of 4'-(4- methanesulphonamidobenzyl) -6-bromo-2 , 2-dimethyl-2 , 3- dihydro-5 ' H-spiro [chromen-4 , 2 ' - [1 , 4] oxazinan] -5' -one (scheme d, step h) (F95)

The compound has been synthesized like in example 20 and has been purified by a chromatographic column using CHC13/MeOH (9.5:0.5) as eluent.

Yield: 47%

¹H-NMR (DMSO) δ: 1.13 (s, 3H); 1.24 (s, 3H); 1.68 (d, IH, J=14.6 Hz); 2.35 (d, IH, J=14.6 Hz); 2.96 (s, 3H); 3.09 (d, IH, J=12.8 Hz); 3.90 (d, IH, J=12.8 Hz); 4.25 (s, 2H); 4.32 (d, IH, J=14.6 Hz); 4.67 (d, IH, J=14.6 Hz); 6.74 (d, IH, J=8.8 Hz); 7.17 (d, 2H, J08.4 Hz); 7.29 (d, 2H, J=8.4 Hz); 7.36 (dd, IH, J=8.8 Hz, J=2.4 Hz); 7.59 (d, IH, J=2.4 Hz).

MS (m/z) : 510 (M⁺, 5%); 184 (100%) Elementary analysis:

C₂₂H_2SN₂O₅SBr C H N

Calc% 51 . 87 4 . 95 5 . 50

Foiand% 51 . 62 5 . 02 5 . 88

Example 22 : Synthesis of 4'-(4- methanesulphonamidobenzyl) -2 , 2-dimethyl-2 , 3-dihydro- 5 'H-spiro[chromen-4, 2 ' - [1 ,4] oxazinane] (scheme d, step h) (F65) The compound has been synthesized like in example 20 and has been purified by means of trsformation into clorhydrate and crystallization by i-PrOH.

Yield: 25%

¹H-NMR (DMSO) δ: 1.25 (s, 6H); 2.32 (d, IH, J=15.5

Hz); 2.64 (d, IH, J=15 Hz); 3.01 (s, 3H) 3.26 (s broad, 2H); 3.53 (s broad, 2H), 3.85-4.1 (m, 2H); 4.15-4.45 (m, 2H); 6.77 (d, IH, J=8.05 Hz); 6.92-7.0 (m, IH); 7.21-7.25 (m 3H); 7.53-7.61 (m, 3H). MS (m/z) : 416 (M⁺, 5%); 184 (100%)

Elementary analysis:

Example 23: Synthesis of 4 ' - (4-methane- sulphonamidobenzyl) -6-bromo-2 , 2-dimethyl-2 , 3-dihydro- 5 ' H-spiro [chromen-4 ,2 ' - [1 , 4] oxazinane] (scheme d, step h) (F134)

The compound has been synthesized like in example 20 and has been purified by means of trsformation into clorhydrate and crystallization by EtOH.

Yield: 45%

¹H-NMR (DMSO) δ: 1.25 (s, 6H); 2.41 (d, IH, J=15

Hz); 2.63 (d, IH, J=15 Hz); 3.01 (s, 3H); 3.20-3.35

(m, IH); 3.56-3.94 (m, 4H), 4.17-4.43 (m, IH); 6.75

(d, IH, J=8.7 Hz); 7.23 (d, 2H, J= 8.2Hz); 7.4 (dd,

IH, J= 8.7, 2.3Hz); 7.62 (d, 2H, J= 8.2Hz); 7.75 (d, IH, J=2.3 Hz) .

Elementary analysis:

Calc% 49 . 68 5 . 31 5 . 27

Found% 49. 50 5 . 0 4 . 95

Example 24: Synthesis of 4 ' - (4-acetamidobenzyl) - 2 , 2-dimethyl-2 , 3-dihydro-5 ' H-spiro [chromen-4 , 2 ' - [1 , 4] oxazinan] -5-one (scheme d, step h) (F163) To a solution of 4 ' - (4-aminobenzyl) -2, 2-dimethyl- 2, 3-dihydro-5 ' H-spiro [chromen-4 , 2 ' - [1, 4] oxazinan] -5- one (lδlmg, 0.514 mmoles) in acetone (3 mL) K₂CO₃ (106.5 mg, l.lδmmoles) and Ac₂O (0.1 mL, 0.514 mmoles) have been added. The reaction mixture has been stirred at AT for 30 minutes. After this period, the solvent has been evaporated and the residual matter treated with AcOEt and washed with water. The dried organic phase has been then evaporated to provide a raw that has been purified by grinding with ethyl ether. Yield: 97%

¹H-NMR (CDCl₃) δ: 1.18 (s, 3H); 1.32 (s, 3H); 1.73 (d, IH, J=14.6 Hz); 2.17 (s, 3H); 2.26 (d, IH, J=14.6 Hz); 3.06 (d, IH, J=12.6 Hz); 3.78 (d, IH, J=12.6 Hz); 4.23-4.44 (m, 3H); 4.88 (d, IH, J=14.2Hz); 6.78-6.94 (m, 2H); 7.16-7.26 (m, 3H); 7.33-7.37 (m, IH); 7.47- 7.51 (m, 2H) .

MS (m/z) : 394 (M⁺, 12%); 146 (100%).

Elementary analysis:

C₂₃H₂₆N₂O₄ C H N

Calc% 70 . 03 6 . 64 7 . 10

Found% 70 . 40 6 . 31 6 . 94

Example 25 : Synthesis of 4 '- (4-acetamidobenzyl) -6- bromo-2 , 2-dimethyl-2 , 3-dihydro-5 ' H-spiro [chromen-4 , 2 ' - [1,4] oxazinan] -5-one (scheme d, step h) (F166) The compound has been synthesized following the same synthetic procedure indicated for example 24. The raw obtained has been purified by grinding with ethyl ether.

Yield: 70%

¹H-NMR (CDCl₃) δ: 1.16 (s, 3H); 1.31 (s, 3H); 1.71 (d, IH, J=14.4 Hz); 2.08-2.27 (m, 4H); 3.06 (d, IH, J=12.6 Hz); 3.73 (d, IH, J=12.6 Hz); 4.23-4.44 (m, 3H); 4.84 (d, IH, J=14.2 Hz); 6.69 (d, IH, J=8.7 Hz); 7.21-7.32 (m, 3H); 7.46-7.53 (m, 2H); 7.69 (s, IH).

Elementary analysis:

C₂₃H₂₅N₂θ₄-3r C H N

Calc% 58 . 36 5 . 32 5 . 92

Found% 58 . 01 5 . 20 5 . 84

Example 26: Synthesis of 4 ' - (4-acetamidobenzyl) - 2 , 2-dimethyl-2 , 3-dihydro-5 ' H-spiro [chromen-4 , 2 ' - [l,4]oxazinane] (scheme d, step h) (F164)

The compound has been synthesed following the same synthetic procedure indicated for example 24. The raw obtained has been purified by transformation into clorhydrate and crystallization by i-PrOH. Yield: 68%

¹H-NMR (DMSO) δ: 1.25 (s, 6H); 2.04 (s, 3H, COCH₃); 2.32 (d, IH, J=15 Hz); 2.65 (d, IH, J=15 Hz); 3.02- 3.08 (m, IH); 3.26 (s broad, 2H); 3.46-3.59 (m, IH); 3.86-4.13 (m, 2H); 4.19-4.34 (m, 2H); 6.75-6.79 (m, IH); 6.92-7.0 (m, IH); 7.20-7.28 (m, IH), 7.52-7.66 (m, 5H) .

Elementary analysis:

Example 27 : Synthesis of 3 ' -benzyl-4 ' -imino-2 ,2- dimethyl-2 , 3-dihydro-2 ' H-spiro [chromen-4 , 5 ' - [l,3]oxazolidin] -2 ' -one (scheme b, step d) (F141) To a solution of carbonyldiimidazole (3.2 mmoles; 515.8 mg) in anhydrous CH₂Cl₂ (6.0 ml) at 0⁰C and in N₂ environment, 4-hydroxy-2, 2-dimethylchroman-4- carbonitrile (589mg, 2.9 mmoles) dissolved in anhydrous CH₂Cl₂ (8.7 ml) has been added drop by drop. The reaction mixture has been stirred at room temperature for 20 minutes. After this period, to the solution benzylamine (310 mg, 2.9 mmoles) has been added and the reaction mixture has been stirred at room temperature for 2 h. After this period, the solution has been washed many times with IN HCl and H₂O, dried and evaporated to p.r. The obtained solid raw has been purified by crystallization by Et₂O. Yield: 27%. 1H-NMR (CDCl₃) δ: 1.39 (s, 3H, CH₃); 1.48 (s, 3H, CH₃); 2.20 (d, IH, J = 15.0 Hz, CH₂); 2.34 (d, IH, J = 15.0 Hz, CH₂); 4.84 (s, 2H, CH₂N); 6.81 - 6.86 (m, 2H, Ar); 7.06 - 7.12 (m, 2H, Ar); 7.21 - 7.49 (m, 5H, Ar); 7.65 (s, IH, NH) ppm.

Melting point: 125 - 130⁰C MS (m/z) : 336 (M⁺, 81%); 292 (-CO₂, 48%);

ELEMENTARY ANALYSIS:

Example 28: Synthesis of 3 ' -benzyl-4 ' -imino-6- bromo-2 , 2-dimethy1-2 , 3-dihydro-2 ' H-spiro [chromen-4 , 5 ' - [l,3]oxazolidin] -2 ' -one (scheme b, step d) (F222)

The product has been synthesized following the same synthetic procedure indicated for example 27. The solid raw has been purified by a chromatographic column using as eluent hexane/AcOEt (7:3).

Yield: 20%.

¹H-NMR (CDCl₃) δ: 1.41 (s, 3H, CH₃); 1.49 (s, 3H, CH₃); 2.17 - 2.39 (m, 2H, CH₂); 4.87 (s, 2H, CH₂N); 6.75 (d, IH, J = 8.9 Hz, Ar); 6.90 (d, IH, J = 2.2 Hz, Ar); 7.32 - 7.45 (m, 6H, Ar) ppm.

Melting point: 126-130⁰C

MS (m/z) : 416 (M⁺, 27%); 372 (M⁺-CO₂, 7%).

Elementary analysis:

C₂oHigθ₃N₂Br C H N

Calc% 57 . 84 4 . 61 6 . 75

Found% 58 . 0 4 . 37 6 . 72

Example 29: Synthesis of 3'-(4- trifluoromethylbenzyl) -4 ' -imino-2 , 2-dimethyl-2 , 3- dihydro-2 ' H-spiro [chromen-4 , 5 ' - [1 , 3] oxazolidin] -2 ' -one (scheme b, step d) (F223)

The product has been synthesized following the same synthetic procedure indicated for example 27. The solid raw has been purified by grinding with Et₂O. Yield: 25%. 1H-NMR (CDCl₃) δ: 1.41 (s, 3H, CH₃); 1.50 (s, 3H, CH₃); 2.22 (d, IH, J = 15.0 Hz, CH₂); 2.37 (d, IH, J = 15.0 Hz, CH₂); 4.93 (s, 2H, CH₂N); 6.76 - 6.91 (m, 3H, Ar); 7.28 - 7.34 (m, IH, Ar); 7.39 (d, 2H, J = 8.0 Hz, Ar); 7.57 (d, 2H, J = 8.0 Hz, Ar) ppm. Melting point: 133 - 136°C. MS (m/z) : 404 (M⁺, 8i8%) ; 360 (-CO₂, 43%) ; Elementary analysis:

C21H19O₃N₂ F3 C H N

Calc% 62 . 37 4 . 74 6 . 93

Found% ^' 62 . 40 4 . 68 7 . 20

Example 30 : Synthesis of 3 ' - (4-aminobenzyl) -4 ' - imino-6-bromo-2 , 2-dimethyl-2 , 3-dihydro-2 ' H- spiro[chromen-4,5 ' - [1 , 3] oxazolidin] -2 ' -one (scheme b, step d) (F226)

The product has been synthesized following the same synthetic procedure indicated for example 27. The solid raw has been purified by a chromatographic column using hexane/AcOEt (6:4) as eluent. Yield: 32%.

¹H-NMR (CDCl₃) δ: 1.38 (s, 3H, CH₃); 1.48 (s, 3H, CH₃); 2.18 (d, IH, J = 15 Hz, CH₂); 2.35 (d, IH, J = 15 Hz, CH₂); 4.75 (s, 2H, CH₂N); 6.68 (d, 2H, J = 8.4 Hz, Ar); 6.74 (d, IH, J = 8.7 Hz, Ar); 6.86 (d, IH, J = 2.3 Hz, Ar); 7.26 - 7.30 (m, 2H, Ar); 7.34 (dd, IH, J = 2.3, 8.7 Hz, Ar) ppm.

Melting point: 155-158°C. MS (m/z): 431 (M⁺, 20%); 387 (-CO₂, 99%).

ELEMENTARY ANALYSIS:

C₂₀H₂₀O₃N₃Br C H N

Calc% 55 . 83 4 . 68 9 . 77

Found% 56 . 01 4 . 77 9 . 80

Example 31: Synthesis of 3'-(4- methanesulphonamidobenzyl) -4 ' -imino -2 , 2-dimethyl-2 , 3- dihydro-2 ' H-spiro [chromen-4 , 5 ' - [1 , 3] oxazolidin] -2 ' -one (scheme d, step h) (F227) To a solution of 3 ' - (4-aminobenzyl) -4 ' -imino-2, 2- dimethyl-2 , 3-dihydro-2 ' ff-spiro [chromen-4 , 5 ' - [1, 3] oxazolidin] -2 ' -one (183 mg, 0.52 mmoles) in anhydrous dioxane (5.5 ml) in nitrogen environment pyridine (0.53 ml) has been added, then the solution has been brought to 0⁰C and methanesulphonylchloride (77.4 mg, 0.68 mmoles) has been added slowly. The solution has been reflux stirred for 1 h, then acidified and extracted with EtOAc. The organic phase has been dried and evaporated. The solid raw has been purified by a chromatographic column using as eluent hexane/AcOEt (1:1). Yield: 32%. 1H-NMR (CDCl₃) δ: 1.40 (s, 3H, CH₃); 1.49 (s, 3H, CH₃); ^2.23 (d, IH, J = 14.8 Hz, CH₂); 2.35 (d, IH, J = 15 Hz, CH₂); 3.01 (s, 3H, CH₃); 4.83 (s, 2H, CH₂N); 6.79 - 6.90 (m, 3H, Ar); 7.20 (d, 2H, J = 8.4 Hz, Ar); 7.24-7.32 (m, IH, Ar); 7.46 (d, 2H, J = 8.4 Hz, Ar) ppm. Melting point: 87 - 9O⁰C.

MS (m/z) : 429 (M⁺, 12%); 385 (-CO₂, 100%);

Elementary analysis:

C₂₁H₂₃O₅N₃S C H N

Calc% 55 . 83 4 . 68 9 . 77

Found.% 56 . 10 4 . 71 9 . 82

Example 32: Synthesis of 3'-(4- methanesulphonamidobenzyl) -4 ' -imino-6-bromo-2 , 2- dimethyl-2 , 3-dihydro-2 ' H-spiro [chromen-4 , 5 ' - [1, 3] oxazolidin] -2 ' -one (scheme d, step h) (F245)

The product has been synthesized following the same synthetic procedure indicated for example 31. The solid raw has been purified by means of precipitation by acOEt/hexane.

Yield: 28%.

¹H-NMR (CDCl₃) δ: 1.38 (s, 3H, CH₃); 1.50 (s, 3H, CH₃); 2.14 (d, IH, J = 15.2 Hz, CH₂); 2.56 (d, IH, J = 15.0 Hz, CH₂); 3.02 (s, 3H, CH₃); 4.89 (s, 2H, CH₂N); 6.69 - 6.81 (m, 3H, Ar); 7.21-7.31 (m, IH, Ar); 7.32- 7.38 (m, IH, Ar); 7.44-7.53 (m, 2H, Ar) ppm.

Melting point: 85 - 88°C.

MS (m/z) : 508 (M⁺, 5%); 465 (M⁺-CO₂, 16%);

Elementary analysis:

C₂iH₂₂θ₅N₃SBr C H N

Calc% 49 . 61 4 . 36 8 . 27

Found% 49 . 38 4 . 69 7 . 95

Example 33 : Synthesis of 3' - (4-acetamidobenzyl) - 4 • -imino-2 , 2-dimethyl-2 , 3-dihydro-2 ' H-spiro [chromen- 4,5'-[l,3]oxazolidin]-2'-one (scheme d, step h) (F246)

To a solution of the suitable imino-oxazolidinone 3 ' - (4-aminobenzyl) -4 ' -imino-2, 2-dimethyl-2, 3-dihydro- 2 ' H-spiro [chromen-4, 5 ' - [1, 3] oxazolidin] -2 ' -one (210mg, 0.6 ramoles) in acetone, K₂CO₃ (118.9 mg; 0.8 mmoles) and acetic anhydride (0.6 mmoles; 0.05 ml) have been added. The reaction mixture has been stirred at room temperature for 30 minutes. After this period, the solvent has evaporated. The residual matter has been treated with AcOEt, washed many times with H₂O, dried and evaporated at low pressure. The raw obtained has been purified by a chromatographic column using hexane/AcOEt (1:4) as eluent. Yield: 45%. 1H-NMR (DMSO) δ: 1.30 (s, 3H, CH₃); 1.44 (s, 3H, CH₃); 2.06 (s, 3H, COCH₃); 2.35 (d, IH, J = 15.0 Hz, CH₂) ; 2.61 (d, IH, J = 15.0 Hz, CH₂) ; 4.68 (s, 2H, CH₂N) ; 6.84 - 6.94 (m, 3H, Ar) ; 7.26 - 7.32 (m, 3H, Ar) ; 7.56 (d, 2H, J = 8.4 Hz, Ar) ppm.

Melting point: 190 - 193⁰C.

MS (m/z) : 393 (M⁺, 14%); 349 (M⁺-COCH₃, 90%);

Elementary analysis:

C22^H23θ4N3 C H N

Calc% 67 .16 5. 89 10. 68

Found% 67 .22 5. 93 10. 32

Example 34 : synthesis of 3 ' - (4-acetamidobenzyl) - 4 ' -imino-6-bromo-2 , 2-dimethyl-2 , 3-dihydro-2 ' H- spiro [chromen-4 ,5 ' - [1 , 3] oxazolidin] -2 ' -one (scheme d, step h) (F247)

The product has been synthesized following the same synthetic procedure indicated for example 33. The solid raw has been purified by a chromatographic column using hexane/AcOEt (3:7) as eluent.

Yield: 17%.

¹H-NMR (CDCl₃) δ: 1.37 (s, 3H, CH₃); 1.48 (s, 3H, CH₃); 2.15 - 2.24 (m, 5H, CH₂, CH₃); 4.83 (s, 2H, CH₂N); 6.75 (d, IH, J = 8.9 Hz, Ar); 6.86 - 6.92 (m, IH, Ar); 7.14 - 7.18 (m, IH, Ar); 7.32 - 7.54 (m, 4H, Ar) ppm.

Melting point: 110 - 113°C.

MS (m/z): 471 (M⁺, 13%); 429 (M⁺-COCH₃, 66%);

Elementary analysis:

^22^2204^3-317 C H N

Calc% 55. 94 4. 69 8. 90

Found% 56. 32 5. 02 8. 58 Example 35: synthesis of 2 ,2-dimethyl-2 , 3-dihydro- 2 ' ff-spiro [chromen-4 , 5 ' - [1 , 3] oxazolidin] -2 ' -one (step d) .

The 4- (aminomethyl) -2 , 2-dimethylchroman-4-ol (1.3Og, 6.33 ramoles) has been solubilised in THF (11 ml) , then this solution has been added drop by drop to a suspension of CDI (1.02 g; 6.33 mmoles) in THF (11 ml) at 0⁰C. The mixture has been stirred at room temperature for 2 hours. After this period the solvent has evaporated; the residual matter has been treated with ethyl acetate and washed many times with diluted

HCl and saturated solution of K₂CO₃. Finally the organic phase has been dried and evaporated. The solid raw has been subject to the next reaction without further purifications.

Yield: 78%.

¹H-NMR (CDCl₃) δ: 1.42 (s, 6H); 2.17 (d, IH, J =

14.6 Hz); 2.46 (d, IH, J = 14.6 Hz); 3.63 (d, IH, J =

8.8 Hz) ; 3.89 (d, IH, J = 8.9Hz) ; 6.83 (d, IH, J = 8.2 Hz); 6.93 - 7.01 (m, IH); 7.22 - 7.29 (m, IH); 7.49

(dd, IH, J= 7.8, 1.3 Hz) ppm.

Example 36 : Synthesis of 6-bromo-2 ,2-dimethyl-2 ,3- dihydro-2 ' H-spiro [chromen-4 , 5 ' - [1 , 3] oxazolidin] -2 ' -one (step d) The product has been synthesized following the same synthetic procedure indicated for example 35. Yield: 84%

¹H-NMR (CDCl₃) δ: 1.41 (s, 6H); 2.14 (d, IH, J =

14.6 Hz); 2.42 (d, IH, J = 14.6 Hz); 3.64 (d, IH, J = 9.0 Hz); 3.86 (d, IH, J = 8.9 Hz); 6.72 (d, IH, J =

8.8 Hz); 7.33 (dd, IH, J = 8.8, 2.4 Hz); 7.58 (d, IH,

J = 2.4 Hz) . Example 37 : synthesis of 3 ' -benzyl-2 , 2-dimethyl- 2 , 3-dihydro-2 ' ff-spiro [chromen-4 , 5 ' - [1 , 3] oxazolidin] - 2 '-one (step f) (F180)

To a suspension of NaH (dispersed in mineral oil at 60%; 13.52 mmoles) in DMF (30 ml), in N₂ environment, 2, 2-dimethyl-2, 3-dihydro-2 ' H- spiro [chromen-4, 5 '- [1, 3] oxazolidin] -2 ' -one (1.0 g,

4.51 mmoles) has been added and the mixture has been stirred at room temperature for about 30 minutes. After this period the mixture has been cooled to 0⁰C and benzyl bromide (1.17 g; 6.8 mmoles) has been added drop by drop, and eventually solubilized in a minimum amount of DMF. The mixture has been brought back to room temperature and stirred for 1 hour. After this period it has been brought to 0⁰C and water has been added. The mixture has been diluted with ethyl acetate and washed with water and a saturated solution of

NaCl. The organic phase is then dried and evaporated.

The raw obtained has been purified by grinding with hexane.

Yield: 50%.

¹H-NMR (CDCl₃) δ: 1.38 (s, 3H); 1.41 (s, 3H); 2.07 (d, IH, J = 14.6 Hz); 2.39 (d, IH, J = 14.6 Hz); 3.40 (d, IH, J = 9.1 Hz); 3.67 (d, IH, J = 9.1 Hz); 4.50 (d, IH, J = 14.8 Hz); 4.58 (d, IH, J = 14.8 Hz); 6.78 - 6.82 (m,lH); 6.86 - 6.94 (m, IH); 7.18 - 7.26 (m, 2H) ; 7.31 - 7.40 (m, 4H) .

Elementary analysis:

C₂₀H₂₁O₃N C H N

Calc% 74 . 28 6 . 55 4 . 33

Found% 74 . 0 6. 27 3 . 98 Example 38 : Synthesis of 3 ' -benzyl-6-bromo-2 ,2- dimethyl-2 , 3-dihydro-2 ' H-spiro [chromen-4 , 5 ' - [l,3]oxazolidin]-2'-one (step f) (F139)

The product has been synthesized following the synthetic procedure indicated for example 37. The solid raw has been purified by chromatography using hexane/AcOEt (7:3) as eluent.

Yield: 63%.

¹H-NMR (CDCl₃) δ: 1.36 (s, 3H); 1.39 (s, 3H); 2.02 (d, IH, J = 14.6 Hz); 2.34 (d, IH, J = 14.6 Hz); 3.39

(d, IH, J = 9.1 Hz); 3.60 (d, IH, J = 9.1 Hz) 4.44 (d, IH, J = 14.8 Hz); 4.63 (d, IH, J = 14.8 Hz) 6.67 (d, IH, J= 8.6 Hz); 7.21 - 7.42 (m, 6H) .

Elementary analysis

C2oH2θθ3NB^r C H N

Calc% 59 . 71 5 . 01 3 . 48

Found% 60 . 02 5 : 24 3 . 64

Example 39: Synthesis of 3 ' - (4-nitrobenzyl) -2,2- dimethyl-2 , 3-dihydro-2 ' H-spiro [chromen-4 , 5 ' - [l,3]oxazolidin]-2'-one (step f) (F155)

The product has been synthesized following the synthetic procedure indicated for example 37. The solid raw has been purified by a chromatographic column using a mixture hexane/AcOEt (1:1) as eluent. Yield: 49%.

¹H-NMR (CDCl₃) δ: 1.39 (s, 3H); 1.41 (s, 3H); 2.08 (d, IH, J = 14.6 Hz); 2.40 (d, IH, J = 14.6 Hz); 3.42 (d, IH, J = 8.9 Hz); 3.71 (d, IH, J = 8.8 Hz); 4.64 (s, 2H); 6.79 - 6.84 (m, IH); 6.88 - 6.95 (m, IH); 7.20 - 7.30 (m, IH); 7.53 (d, 2H, J = 8.6 Hz); 7.69 (d, IH, J= 8.8 Hz); 8.27 (d, 2H, J= 8.6 Hz). Melting point: 78-81°C. Elementary analysis:

C20H20O5N2 C H N

Calc% 65 . 21 5 . 47 7 . 60

Found% 65 . 07 5 . 69 7 . 29

Example 40 : Synthesis of 3 ' - (4-nitrobenzyl) -6- bromo-2 , 2-dimethyl-2 , 3-dihydro-2 ' H-spiro [chromen-4 , 5 ' - [l,3]oxazolidin]-2'-one (step f) (F178)

The product has been synthesized following the synthetic procedure indicated for example 37. The solid raw has been purified by a chromatographic column using a mixture hexane/AcOEt (1:1) as eluent. Yield: 40%.

¹H-NMR (CDCl₃) δ: 1.38 (s, 3H); 1.40 (s, 3H); 2.04 (d, IH, J = 14.6 Hz); 2.38 (d, IH, J = 14.6 Hz); 3.44 (d, IH, J = 9.2 Hz); 3.68 (d, IH, J = 9.0 Hz); 4.56 (d, IH, J = 15.2 Hz); 4.71 (d, IH, J = 15.4 Hz); 6.70 (d, IH, J = 7.7 Hz); 7.26 - 7.33 (m, 2H); 7.55 (d, 2H, J= 8.6 Hz); 8.29 (d, 2H, J= 8.6 Hz).

Elementary analysis:

Example 41 : Synthesis of 3'-(4- trifluoromethylbenzyl) -6-bromo-2 , 2-dimethyl-2 , 3- dihydro-2 ' H-spiro [chromen-4 , 5 ' - [1 , 3] oxazolidin] -2 ' -one (step f) (F172)

The product has been synthesized following the synthetic procedure indicated for example 37. The solid raw has been purified by grinding with ethyl ether. Yield: 61%.

¹H-NMR (CDCl₃) δ: 1.38 (s, 3H) ; 1.39 (s, 3H) ; 2.04

(d, IH, J = 14.6 Hz) ; 2.36 (d, IH, J = 14.6 Hz) ; 3.40

(d, IH, J = 9.2 Hz) ; 3.63 (d, IH, J = 9.2 Hz) ; 4.53

(d, IH, J = 15.2 Hz) ; 4.65 (d, IH, J = 15.0 Hz) ; 6.69 (d, IH, J = 9.5 Hz) ; 7.26 - 7.31 (m, 2H) ; 7.48 (d, 2H, J = 7.9 Hz) ; 7.68 (d, 2H, J = 8.0 Hz) ppm.

Melting point: 138-141°C.

Elementary analysis:

C₂iH₁₉O₃NF₃Br C H N

Calc% 57. 45 5. 08 6. 09

Found% 57. 64 4. 98 6. 25

Example 42 : Synthesis of 3 ' - (4-aminobenzyl) -2 , 2- dimethyl-2 , 3-dihydro-2 ' H-spiro [chromen-4 , 5 ' - [l,3]oxazolidin] -2' -one (scheme d, step h) (F157)

To a solution of 3 ' - (4-nitrobenzyl) -2, 2-dimethyl- 2, 3-dihydro-2 ' H-spiro [chromen-4, 5 ' - [1, 3] oxazolidin] - 2 ' -one (645mg, 1.75mmoles) dissolved in methanol (19mL) coal (50mg) and an amount catalytic of FeCl₃ have been added. The reaction mixture has been reflux heated and hydrated hydrazine (1.4mL, 29.7 mmoles) has been added. The mixture has been reflux stirred for 12h. After this period, the suspension has been filtered on celite and the solvent evaporated. The residual matter has been purified by grinding in i- Pr₂O.

Yield: 92%.

¹H-NMR (CDCl₃) δ: 1.37 (s, 3H); 1.39 (s, 3H); 2.03 (d, IH, J = 14.6 Hz); 2.34 (d, IH, J = 14.6 Hz); 3.36 (d, IH, J = 9.2 Hz); 3.62 (d, IH, J = 9.2 Hz); 4.96 (d, IH, J = 14.4 Hz); 4.45 (d, . IH, J = 14.4 Hz); 6.67 (d, 2H, J = 8.4 Hz); 6.78 (d, IH, J = 8.2 Hz); 6.84 - 6.92 (m, IH) ; 7.12 (d, 2H, J = 8.4 Hz) ; 7.20 - 7.26 (m, 2H) .

Elementary analysis:

C20H22O3N2 C H N

Calc% 70. 99 6. 55 8. 28

Found% 70. 75 6. 37 8. 09

Example 43 : Synthesis of 3 ' - (4-acetamidobenzyl) -6- bromo-2 , 2-dimethyl-2 , 3-dihydro-2 ' H-spiro [chromen-4 , 5 ' - [l,3]oxazolidin]-2'-one (step h) (F279)

The product has been synthesized following the same synthetic procedure indicated for example 33. The solid raw has been purified by a chromatographic column using as eluent hexane/AcOEt (3:7).

Yield: 50%.

¹H-NMR (CDCl₃) δ: 1.36 (s, 3H); 1.37 (s, 3H); 2.02 (d, IH, 14.6 Hz); 2.17 (s, 3H); 2.31 (d, IH, J = 14.6 Hz); 3.39 (d, IH, J = 9.3 Hz) 3.60 (d, IH, J = 9.1 Hz); 4.41 (d, IH, J = 14.8 Hz) 4.54 (d, IH, J = 14.8 Hz) ; 6.67 (d, 2H, J = 8.7 Hz) 7.20 - 7.29 (m, 3H); 7.51- 7.61 (m, 2H) ppm.

Melting point: 79-82°C.

Elementary analysis:

C₂₂H₂₃O₄N₂Br C H N

Calc% 57. 53 5. 05 6. 10

Found% 57. 15 5. 31 5. 92

Example 44 : Synthesis of 3 ' - (4-methane- sulphonamidobenzyl) -2 , 2-dimethyl-2 , 3-dihydro-2 ' H- spiro[chromen-4, 5' - [1 , 3] oxazolidin] -2' -one (step h) (F276) The product has been synthesized following the same synthetic procedure indicated for example 31. The solid raw has been^' purified by grinding with ethyl ether .

Yield: 95%.

¹H-NMR (DMSO) δ: 1.32 (s, 6H); 2.22 (d, IH, J = 14.8 Hz); 2.35 (d, IH, J = 14.7 Hz); 2.99 (s, 3H); 3.52 (d, IH, J = 9.5 Hz); 3.66 (d, IH, J = 9.3 Hz); 4.42 (s, 2H); 6.78 (d, IH, J = 8.2 Hz); 6.88 - 6.96 (m, IH); 7.12 -7.33 (m, 6H) ppm.

Melting point: 120-122°C.

ELEMENTARY ANALYSIS:

C₂₁H₂₄O₅N₂S C H N

Calc% 60 . 56 5 . 81 6 . 73

Found% 60 . 32 5 . 77 6 . 89

Example 45 : Synthesis of 3 ' -benzyl-2 ,2-dimethyl- 2 , 3-dihydro-2 ' H, 4 ' H-spiro [chromen-4 , 5 ' -

[l,3]oxazolidin]-2' , 4 ' -dione (step f) (F298)

To a solution of 3 ' -benzyl-4 ' -imino-2, 2-dimethyl- 2, 3-dihydro-2 ' iϊ-spiro [chromen-4 , 5 ' - [1, 3] oxazolidin] - 2'-one (74mg, 0.22 moles) in THF, HCl IN (0.4 ml) has been added at 0⁰C. It is stirred at room temperature for about 2 hours. After this period the solvent has evaporated and the acid aqueous phase is extracted with ethyl acetate. The organic phase is dried and evaporated. The raw corresponding to the desired product is chopped with ethyl ether. Yield: 30%.

¹H-NMR (CD₃OD) δ: 1.41 (s, 3H); 1.55 (s, 3H); 2.54 (d, IH, J = 15.2 Hz); 2.74 (d, IH, J = 15.2 Hz); 5.09 (s, 2H); 6.87 - 6.98 (m, 3H); 7.36 - 7.50 (m, 5H) ppm. Melting point: 148-150⁰C. Elementary analysis:

C₂₀Hi₉O₄N C H N

Calc% 71 . 20 68 4 . 15

Found% 71 . 0 5 . 42 3 . 98

Example 46 : Synthesis of 3 ' -benzyl-6-bromo-2,2- dimethyl-2 , 3-dihydro-2 ' H₁4 ' H-spiro [chromen-4 , 5 ' - [l,3]oxazolidin]-2' , 4 ' -dione (step f) (F299)

The product has been synthesized following the same synthetic procedure indicated for example 45. The solid raw has been purified by means of AcOEt/hexane precipitation. Yield: 73%. ^{) I}T

¹H-NMR (CDCl₃) δ: 1.36 (s, 3H); 1.49 (s, 3H); 2.13 (d, IH, J = 15.0 Hz); 2.56 (d, IH, J = 15.0 Hz); 4.79 (s, IH); 4.80 (s, IH); 6.74 - 6.78 (m, 2H); 7.32 (d, IH, J= 2.4 Hz); 7.36 - 7.46 (m, 5H) ppm. Melting point: 148-151°C.

MS (m/s) : 416 (M⁺, 97%); 266 (100%).

Elementary analysis:

C₂oHi₈θ₄NBr C H N

Calc% 57 . 71 4 . 36 3 . 36

Found% 58 . 02 5 . 05 3 . 34

Experimental results

All the experimental procedures described hereafter have been carried out according with the guidelines of CEE Directive 86-609, relative to experimentation on animals

Male albine mice of the Wistar type (260-350 g) have been treated with an intraperitoneal injection (i.p.) of the drug of reference, i.e. with diazoxide (40 mg/Kg) , as well as with F 95 (40 mg/Kg) , or F 134 (40 mg/Kg) or F 163 (40 mg/Kg), which are molecules selected according to the invention. Both the diazoxide and the molecules according to the invention, have been dissolved in the dimethylsulphoxide (DMSO) vehicle. A mice control group has been treated instead with an i.p. injection only with the DMSO vehicle. Two hours after supplying the drugs or the vehicle, the animals have been anaesthetized with sodium pentobarbital (100 mg/Kg i.p.) and treated with heparin (100 UI, i.p.). After thoracotomy, the heart has been explanted and immersed in a Krebs solution (mM composition: NaHCO₃ 25.0, NaCl 118.1, KCl 4.8, MgSO₄ 1.2, CaCl₂.2H₂O 1.6, KH₂PO₄ 1.2, glucose 11.5) cooled to 4° C and bubbled with a gaseous mixture of O₂ (95%) and CO₂ (5%) . Within two minutes from the explantation, the heart has been perfused at fixed pressure (70-80 mm/Hg) through catheterization of the ascending portion of the aortic arch with Krebs solution, heated to 37° C and bubbled with a gaseous mixture of O₂ (95%) and CO₂ (5%), and housed in a thermostated chamber at 37° C, according to the Langendorff method. For determining the systolic pressure developed by the left ventricle

(LVDP) and the cardiac frequency (HR) , a balloon of latex rubber, connected to a pressure transducer, has been inserted through the left ventricle mitralic valve, and then it has been filled with water up to reaching a diastolic pressure of 5-10 mm/Hg. The functional parameter of inotropism has been calculated as product of frequency and pressure (RPP = LVDP x HR) . To avoid that the correct analysis of the functional parameter is influenced by a physiological decrease of contractile activity reconducible to the long reperfusion period (where the isolated organ could suffer indirectly - or in any case not exclusively - from ischemia) , in the data analysis data in addition to RPP value observed at the last minute of the reperfusion period (RPP-120' ) , also an RPP value detected at the 30^th min of the reperfusion period (RPP-30' ) has been considered. The parameters RPP-120' and RPP-30' have been expressed as % with respect RPP value recorded at the last minute of the pre-ischemic period. The coronary flow has been monitored volumetrically, as ml of perfusate exiting from the isolated heart for minute. After 30 min of pre-ischemic period, necessary to allow the heart to reach a situation of balance, the perfusion has been stopped and the heart has been exposed to a period of 30 min of global ischemia. The hearts that in the pre- ischemic period have shown arrhythmias or have not achieved a situation of balance in the functional parameters of LVDP and HR, have been discarded by the experiment. At the end of the ischemic period, the perfusion has been reactivated and the heart has been then exposed to a period of 120 min of reperfusion. With method spectrophotometric, lactate dehydrogenase enzyme (LDH, a biochemical marker of ischemic injury) has been measured in the perfusate collected in the last 5 min of the pre-ischemic period and in that collected every 5 min, during all the reperfusion step. The amount of LDH released has been expressed as enzymatic mU released in the 120 min of reperfusion (by subtracting the possible low amount detected in the pre-ischemic period) , calculated by means of analysis of the AUC (area under the curve in the cartesian chart of the LDH amounts detected at the predetermined intervals VS time) and compared to 1 g of weight of the heart. At the end of the reperfusion period, the hearts have been removed by the chamber and the left ventricle has been cut into transversal slices of about 1 mm of thickness, which have been immersed for 20 min in a 10% aqueous solution of triphenyltetrazolium chloride (TTC) and then in a 20% aqueous solution of formaldehyde. After 24 hours, the ventricular slices have been photographed and analyseste for showing the necrotic areas damaged by ischemia (white or pale pink coloured) and the sound areas (coloured intense red by the TTC) and to calculate then the % of ischemic area with respect to the total myocardial area. Vehicle The control isolated hearts (animals pretreated with the vehicle) , subject to the ischemia/reperfusion cycle (30 min/120 min) according to the described protocol, have allowed measuring a substantive damage induced by the 30 min of global ischemia.

In particular, the functional parameter of inotropism has result significantly compromised in the post ischemic period (RPP-120' = 23; RPP-30' = 23) . During the 120 min of reperfusion the hearts of control have released 8766 mU/g of LDH. At the end of the reperfusion period, the hearts of control have shown the presence of wide zones of tissue damaged by the ischemia and the % of ischemic area with respect to total myocardial area has risulted the same as 35.

Diazoxide The hearts isolated from the animals pretreated with the drug of reference diazoxide, and subject to the cycle ischemia/reperfusion (30 min/120 min) according to the described protocol, have shown a functional parameter of post-ischemic inotropism higher than the control (RPP-120' = 34; RPP-30' = 86). During the 120 min of reperfusion these hearts have released 2091 mU/g of LDH, a value less than the control. At the end of the reperfusion period, the hearts of the animals pretreated with the diazoxide have shown a presence of zones of tissue damaged by the ischemia reduced with respect to the control, with a % of ischemic area with respect to total myocardial area the same as 17. F 95 The hearts isolated from the animals pretreated with F 95, subject to the cycle ischemia/reperfusion (30 min/ 120 min) according to the described protocol, have shown a functional parameter of post-ischemic inotropism higher than the control (RPP-120' = 57; RPP-30' = 75) . During the 120 min of reperfusion these hearts have released 2992 mU/g of LDH, a value less than the control. At the end of the reperfusion period, the hearts of the animals pretreated with F 95 have shown a presence of zones of tissue damaged by the ischemia reduced with respect to the control, with a % of ischemic area with respect to total myocardial area the same as 13. F 134 The hearts isolated from the animals pretreated 7

- 55 - with F 134, subject to the cycle ischemia/reperfusion (30 min / 120 min) according to the described protocol, have shown a functional parameter of post- ischemic inotropism higher than the control (RPP-120' = 77; RPP-30' > 100) . During the 120 min of reperfusion these hearts have released 6870 mU/g of LDH, a value less than the control. At the end of the reperfusion period, the hearts of the animals pretreated with F 134 have shown a presence of zones of tissue damaged by the ischemia reduced with respect to the control, with a % of ischemic area with respect to total myocardial area the same as 14. F 163 The hearts isolated from the animals pretreated with F 163, subject to the cycle ischemia/reperfusion (30 min/ 120 min) according to the described protocol, have shown a functional parameter of post-ischemic inotropism higher than the control (RPP-120' = 43; RPP-30' > 100) . During the 120 min of reperfusion these hearts have released 5978 mU/g of LDH, a value less than the control. At the end of the reperfusion period, the hearts of the animals pretreated with F163 have shown a presence of zones of tissue damaged by the ischemia reduced with respect to the control, with a % of ischemic area with respect to total myocardial area the same as 20.

The results relative to the tests above described are given in figures from A to C.

In figure A the recover levels of the contractile cardiac function, recorded at the 30° minute of reperfusion after a period of global ischemia of 30 minutes are shown, in hearts isolated from Wistar mice pretreated with the vehicle, with the diazoxide (Diaz) drug of reference or with three of the molecules according to the invention (F 95, F 134 and F 163) . The contractile function, shown as RPP (product of the cardiac frequency multiplied per the pressure of the left ventricle) is expressed as a ratio with respect to the values of RPP recorded in the pre-ischemic period.

In figure B the levels are shown of the enzyme lactate dehydrogenase (LDH) released during the 120 minutes of reperfusion subsequent to a period of global ischemia of 30 minutes, from hearts isolated from Wistar mice pretreated with the vehicle, with the diazoxide (Diaz) drug of reference or with three of the molecules according to the invention (F 95, F 134 and F 163) . The amount of LDH released is expressed in mU for Ig of weight of the heart.

In figure C the ratio of myocardial area damaged by the global ischemia with respect to total area is shown. The ischemic zones have been shown by the observation of slices of left ventricle, obtained from hearts isolated from Wistar mice pretreated with the vehicle, with the diazoxide (Diaz) drug of reference or with three of the molecules according to the invention (F 95, F 134 and F 163), subject to a cycle of 30 minutes of ischemia and 120 minutes of reperfusion and then exposed to treatment with triphenyltetrazolium chloride.

Claims

- 58 -

CLAIMS 1. Eterocyclic compound characterised in that it has the following general formula (X) : wherein

— * represents a chiral center.

— R₀ is selected from the group comprised of: a carbonyl or thiocarbonyl group, an alkyl group (methylene, ethylene, or methylcarbonyl or methyl- thiocarbonyl group.

— Y is selected from the group comprised of: a CH₂ group/ a C=O group, a C=S group, a C=NH group.

— A is selected from the group comprised of: a - CONH- group, a -COO- group, a -CO- group, an alkyl group (C1-C3), alkylcarbonyl group, a carbonyl group, a thiocarbonyl group, an alkylthiocarbonyl group, a sulphonic group, an alkylsulphonic group.

— Ri is selected from the group comprised of: a hydrogen atom, an alkyl group, (methyl, ethyl, propyl, isopropyl, butyl, iso-butyl or tert-butyl, an alkoxy group (methoxy, ethoxy, n-, iso- propyloxy) , an halide atom (F, Cl, Br, I) , a trifluoromethyl group, a cyanide group, a nitro group, a hydroxy group, an amine group, an alkylamine group, an alkylamide group (acetamide, trifluoroacetamide, propionamide) or - 59 - alkylsulphonamide (methanesulphonamide, ethane- sulphonamide) .

— R₂ is selected from the group comprised of: a hydrogen atom, an alkyl group with C1-C4 carbon atoms (methyl, ethyl, propyl, isopropyl, butyl, isobutyl or terbutyl) , a carboxyl group, an alkoxy group (methoxy, ethoxy, n-propyloxy, iso- propyloxy) , an halide atom (F, Cl, Br, I), a cyanide group, a nitro group, a trifluoromethyl group, a hydroxy group, a thioalkyl group (Cl, C2, C3) ; an amine group of NR3R4 type, where R3 and R4 can be indifferently a hydrogen atom, an alkyl, alkylsulphonic (methane-sulphonic, ethane- sulphonic) , acyl (acetyl, propionyl) , trifluoro- alkyl group.

2. A compound, according to claim 1 wherein: Ro is a -CH₂CO- group and Y is a-CH₂~ group.

3. A compound, according to claim 1, wherein Ro is a -(CH₂) 2- group and Y is a group for the -CH₂- type.

4. A compound, according to claim 1, wherein Ro is a C=O group and Y is selected from the group comprised of: -CH₂-, C=O, C=NH.

5. A compound, according to one of the claims from 2 to 4, wherein Y is nitrogen substituted with a substituting group selected from the group comprised of: an alkyl group, an acyl group, a benzyl group.

6. A compound, according to claim 5, wherein said substituting group is in turn substituted by alkyl, amine or alkylamine (C1-C4), amide, N- alkylsulphonamide or N-alkylamide, halide (Cl, Br, F, I), alkyl halide (trifluoromethyl) groups, or - 60 - electron withdrawing groups (nitro or cyanide) .

7. A compound, according to claims from 1 to 6, wherein said general formula (X) comprises at least one among: — enantiomer (R) ,

— enantiomer (S) ,

— racemic mixture of said enantiomer (R) and of said enantiomer (S) .

8. A pharmaceutical composition for treatment of biological tissues involved with ischemic episodes characterised in that it comprises a measured amount of at least one compound having general formula (X), according to claims from 1 to 7.

9. A composition, according to claim 8, characterised in that it induces a ischemic preconditioning through the activation of KATP channels .