CZ2000937A3 - Bisethers of 1-oxa, aza and thianaphthalen-2-one functioning as inhibitors of phospholamban and pharmaceutical preparations containing thereof - Google Patents

Abstract

The therapeutically active compound of formula (I) or (II) in which it is present R 1 is hydrogen, alkyl, alkenyl, aryl, arylalkyl, hydroxyalkyl. haloalkyl, alkoxy, COR 10, CONR 10 R 11, OR 10, S (O) m R 10, NR 10 COR 6, or NR 10 R 11 wherein R 10 is hydrogen, alkyl, alkenyl, aryl, arylalkyl, hydroxyalkyl, haloalkyl, alkoxy or hydroxy and R 1 is hydrogen, alkyl, aryl, arylalkyl, alkoxy, aryloxy, hydroxy or acyl, or in the case of X is NR n, R 1 may also be carboxyalkyl; R 6 is hydrogen, alkyl, alkenyl, aryl, alkyl; R 2 and R 7 are hydrogen, alkyl, alkyl, arylalkyl, alkenyl, COR 10, CONR 10 R 11, halogen, CF 3, NU or CN; R3 is hydrogen, alkyl, aryl or aralkyl; Is alkyl or substituted alkyl, m is 0-2 and n is 1-3; Y is O, NR n, or S; X is O, NR 11 or S; R4, R5, R5 and R9 means an independent of the following groups (a), (b), ( c), (d), (e), (f), (g), or in the case where X is NR n, may also R 1, R 5, R 8 and R 9 independently represent HOOC-, R 12 OOC-, H2NCCCCO- or HOHNCO-, wherein R12 is alkyl arylalkyl or aryl, and their pharmaceutically acceptable salts and esters. Compounds have phospholamban inhibitory activity and are suitable for the treatment of heart failure.

Description

Ether-Oxa-, aza- and thianaphthalen-2-one bisethers as phospholambane inhibitors and pharmaceutical compositions

Technical field:

The present invention relates to novel therapeutically active compounds and salts and esters thereof, as well as to novel intermediates. The invention also relates to pharmaceutical compositions containing these compounds as active ingredients. The compounds of the invention have phospholambane inhibiting properties and are useful in the treatment of heart failure and myocardial paralysis.

SUMMARY OF THE INVENTION:

The compounds of the present invention have the structure represented by formula (I) or (II):

in which

R ¹ is hydrogen, alkyl, alkenyl, aryl, arylalkyl, hydroxyalkyl, haloalkyl, alkoxy, COR ₁₀ , CONR ₁₀ Rh, OR ₁₀ , S (O) _m R ₁₀ , NR ₁₀ COR 11, or NR 10q H, wherein R ₁₀ is hydrogen, alkyl, alkenyl, aryl, arylalkyl, hydroxyalkyl, haloalkyl, alkoxy or hydroxy, and R _n is hydrogen, alkyl, aryl, • · ·· ···· ". ♦ ···· ··· from ··· · · · Arylalkyl, alkoxy, aryloxy, hydroxy or acyl, or when X is NR _1b , R ₁ may also be carboxyalkyl,

R ⁶ is hydrogen, alkyl, alkenyl, aryl, arylalkyl,

R and R are hydrogen, alkyl, aryl, arylalkyl, alkenyl, COR ₁₀ ,

CONR ₁₀ R ₁₁ , halogen, trifluoromethyl, nitro or cyano, wherein R ₁₀ and R ₁₀ are as defined above,

R ₃ is hydrogen, alkyl, aryl or arylalkyl,

A is alkyl or substituted alkyl, m is 0-2 and n is 1-3,

Y represents O, S or NR h, wherein R _n is the same as defined above,

X represents O, S or NR _N, wherein R is the same _thickness as defined above, R _4, R _5, R _g and R ₉ are each independently one of the following groups:

or in the case where X is NR ₁₁ , R ₄ , R ₅ , R ₈ and R ₉ independently can also be HOC-, R ₁₂ OOC-, H ₂ NCO- or HOHNCO-, wherein R ₁₂ is alkyl, arylalkyl or aryl, and wherein each aryl radical as defined above, alone or as part of another group, may be substituted, and pharmaceutically acceptable salts and esters thereof.

In one class of preferred compounds and pharmaceutically acceptable salts and esters thereof are compounds of formula (I) wherein R 2 is hydrogen. In a subclass of this class of compounds and pharmaceutically acceptable salts and esters thereof, R 1 is hydrogen, C 1-6. ₆ alkyl, c _{2. 6} alkenyl, C ₆ . ₁₀ aryl, C ₇ . ₁₂ arylalkyl, C 1-6 hydroxyalkyl, C 1-6 alkyl; ₆ haloalkyl or C _1-6 alkoxy. In the group of this subclass of compounds and pharmaceutically acceptable salts and esters thereof, Y is O or S, preferably O; and X is O. In another group of this subclass of compounds and pharmaceutically acceptable salts thereof, they are pharmaceutically acceptable. Salts and esters Y is O or S and X is NR 11, where R 11 is hydrogen, C i. ₆ alkyl, C ₆ _ ₁₀ aryl, C ₇ _i ₂ arylalkyl, C. ₆ alkoxy, C _{6. 10} aryloxy, hydroxy, C ₆ alkanoyl or Cj.ékarboxyalkyl. In a subgroup of this group of compounds and pharmaceutically acceptable salts and esters thereof, R3 is hydrogen, C _'6 alkyl, C _{6. 10} aryl or C _{7th 12} arylalkyl, preferably C1-6 alkyl. ₆ alkyl, most preferably methyl. In the family of these subgroups of compounds and their pharmaceutically acceptable salts and esters, A is preferably a straight chain or branched C 1. ₄ alkylene.

In another preferred class of compounds and pharmaceutically acceptable salts and esters thereof, the compounds have the formula (II) wherein R 7 is hydrogen. In a subclass of this class of compounds and their pharmaceutically acceptable salts and esters, R 6 is hydrogen, C 1-6. ₆ alkyl, C _{6. 10} aryl, C ₇ . ₁₂ arylalkyl; and n is 1, 2 or 3, preferably 1 or 2. In the group of this subclass of compounds and pharmaceutically acceptable salts and esters thereof, Y is O or S, preferably O; and X is O. In another group of this subclass of compounds and pharmaceutically acceptable salts and esters thereof, Y is O or S, preferably O; and X is NR 11, wherein R 11 is hydrogen, C 1-6. ₆ alkyl, C _{6. 10} aryl, C ₇ _ ₁₂ aralkyl, C ₆ alkoxy, C _{6. 10} aryloxy, hydroxy, Cj. ₆ alkanoyl or C _1-6 carboxyalkyl. In a subgroup of this group of compounds and pharmaceutically acceptable salts and esters thereof, A is preferably a straight chain or branched C ₁ Series. ₄ alkylene.

Each aryl moiety in each of these preferred classes of compounds, alone or as part of another group, may be substituted with 1 to 3, preferably 1 or 2, most preferably one fluorine, chlorine, bromine, iodine, trifluoromethyl, amino, C 1-4 alkyl, C ₁ . ₄ alkoxy, phenyl, naphthyl, halogenofenylem, halogenonaftylem, benzyl, phenethyl, halogenobenzylem, halogenofenethylem, naphthylmethyl, naphthylethyl, C _{4th 7} cycloalkyl, C _{1st 4} alkyl (C _4.7) cycloalkyl, hydroxy, mono (C _1.4) alkylamino, diíCXJalkylaminoskupinou, C. ₆ alkanoylamino, phenylcarbonylamino, naftylkarbonylaminoskupinou, nitro, cyano, thiol, or C _{first 6} alkylthio.

Description of the drawings:

• 0 • ·

0 0 0 • 0

GIANT. 1 depicts the effect of the compound of Example 1c (50 and 100 µM) on the Ca ²⁺ removal rate in SR SR bladders of the heart muscle.

GIANT. 1B shows the effect of the compound of Example 1c (50 and 100 µM) on the Ca ²⁺ removal rate in SR skeletons of solid skeletal muscle.

GIANT. 2A depicts the development of myocardial paralysis and subsequent decline in left ventricular systolic pressure.

GIANT. 2B depicts complete inhibition of myocardial paralysis by the compound of Example 8g.

The compounds of the present invention can be prepared from 1,3-dihydroxy-substituted heteroaromatics by alkylation of the dihydroxy compounds with suitable alkylating agents, for example, chloroacetonitrile or bromoacetic acid ester according to the following Scheme 1 wherein R 1, R ₂ , R 3, X and Y are as defined above. R 'is a hydroxyl protecting group such as methyl, benzyl or tetrahydropyranyl.

SCHEME I

Cl '

N

'R'

K ₂ CO ₃ , DMF

60-120 ° C, 1-5h

K ₂ CO ₃ , DMF 60-120 ° C, 1-5h

• · • ·

The cyano compound (IV) described above is used to prepare 1,2,4-oxadiazole and 1,2,4-thiadiazole derivatives using the method described in J. Med. Chem. 1996, 39, 5228-5235.

The syntheses are shown in Scheme 2, wherein R 1, R ₂ , R ₃ , X and Y are the same as defined above.

SCHEME 2

NH 3 OHXHCl, DMSO

R2 • 4

4444 ø 4 4 »4 4 4 4

Other heterocyclic compounds such as R ₄ , R ₅ , R ₈ and R ₉ are prepared according to the procedures described in Bioorg. Copper. Chem. Lett., 1994, 4, 45-50.

The dihydroaromatic compounds (III) are made using methods known from the literature. Coumarins (XIV), (XVI) and (XX) are produced using Knoevenagel condensation, or von Pechmann reaction, presented in Schemes 3 and 4, wherein R _15, R ₂ and R ₃ are as defined above,

Z is alkyl, aryl, arylalkyl or alkenyl; and R 'is a hydroxyl protecting group such as methyl, benzyl or tetrahydropyran 1.

SCHEME 3

Zn (Hg) -HCl

T

(XIV) · · toto · * · ♦ ♦ ♦ ♦ ♦ ♦ ♦ · · · · · to to to to · ···· ·· ··· ·· this

SCHEME 4

R3

HO '' OH

Λ-RI

HCl, EtOH

OH R3

(XVI)

Pipendin x DBU

(XVIII)

Quinolines are prepared by a Knorr reaction as described in Scheme 5, wherein R

R _n and R ₃ are as defined above, X is halogen.

Φ ·· • Φ Φ 4 • · 4

ΦΦ *** · »Φ» · · · · · · ·

ΦΦ ΦΦΦ ΦΦ ΦΦ

SCHEME 5

^{+ R1 R1} ο

(XXI)

140-160 ° C

R1 (XXII)

(XXIV) (XXIV)

Pyridine χ HCl

Η

(XXV) fr frfrfrfr fr fr fr fr fr fr fr fr fr fr fr fr fr fr fr fr fr fr fr fr fr fr fr fr fr fr fr fr fr fr fr fr fr fr fr fr fr fr fr fr fr fr fr fr fr fr fr fr fr fr fr fr fr fr fr fr fr fr fr fr fr fr fr fr fr fr fr fr

The cyclic compounds (II) may be prepared correspondingly from compound (XXXI), which may be prepared according to Scheme 6, wherein R _{2 and} R ₆ are as defined above, R 1 is a hydroxyl protecting group such as methyl, benzyl or tetrahydropyranyl.

SCHEME 6 fr

OH

R6 [CHJn

(XXVII) (XXVII)

H

(XXXI)

99

9 9 9

9 9

9 9 9

9 9

9999 • 9999

9 9,999 · · · · 9

9 * 9

9 9

9 9 9

9 9 9

9 9 9

9 '9-9

The quinoline cyclic compounds (II) can be prepared by the corresponding procedure of (XXVI) using Scheme 5.

Salts and esters of the compounds may be prepared by known methods, if applicable. Physiologically acceptable salts are suitable as active drugs, and alkali or alkaline earth metal salts are preferred, physiologically acceptable esters are also useful as active drugs. Examples are esters with aliphatic or aromatic alcohols.

The term "alkyl" as used herein by itself or as part of another group includes straight, branched, or cyclized radicals of up to 18 carbon atoms, preferably 1 to 8 carbon atoms, most preferably 1 to 4 carbon atoms. The term "lower alkyl" as used herein by itself or as part of another group includes straight, branched, or cyclized radicals of 1 to 7 carbon atoms, preferably 1 to 4 carbon atoms, most preferably 1 to 2 carbon atoms. Particular examples of alkyl and lower alkyl radicals are methyl, ethyl, propyl, isopropyl, butyl, tert-butyl, pentyl, cyclopentyl, hexyl, cyclohexyl, octyl, decyl, and dodecyl including their various branched isomers.

The term "acyl" as used herein by itself or as part of another group refers to an alkylcarbonyl or alkenylcarbonyl group, wherein the alkyl and alkenyl groups are as defined above.

The term "aryl" as used herein by itself or as part of another group refers to a monocyclic or bicyclic group containing from 6 to 10 carbon atoms in the ring moiety. Particular examples of aryl groups are phenyl, naphthyl and the like. "Aroyl" refers accordingly to an arylcarbonyl group.

The term "alkoxy," as used herein by itself or as part of another group, includes an alkyl group, as defined above, attached to an oxygen atom. "Aryloxy-" refers accordingly to an aryl group attached to an oxygen atom.

The term "substituted" as used herein in connection with various radicals refers to halogen substituents such as fluorine, chlorine,

0 * »0 ·· * 00 00 • ·· 0 0 00 0 00« «0 0 000 000« 000

Π0 0 0 00 0 000 000 0

0 0 0 0 0 0 * 00

0000 · 0 * ·· * 0 0 0 bromo, iodo or trifluoromethyl, amino, alkyl, alkoxy, aryl, alkyl-aryl, halo-aryl, cycloalkyl, alkylcycloalkyl, hydroxy, alkylamino, alkanoylamino, arylcarbonylamino, nitro, cyano, thio, thio, cyano, thio, cyano alkylthiosubstituted.

The "substituted" groups may contain 1 to 3, preferably 1 or 2, most preferably 1 of the aforementioned substituents.

The compounds of the invention may be administered to a patient in therapeutically effective amounts, which usually range from about 0.1 to 500 mg per day, depending on the age, weight, condition of the patient, route of administration, and phospholambane inhibitor used. The compounds of the invention may be formulated into dosage forms using the principles known in the art. They may be administered to the patient as such, or in combination with suitable pharmaceutical excipients in the form of tablets, dragees, capsules, suppositories, emulsions, suspensions or solutions. The selection of suitable formulation ingredients is routine to those of ordinary skill in the art. It will be appreciated that suitable carriers, solvents, gel-forming additives, dispersion-forming additives, antioxidants, colorants, sweeteners, wetting compounds and other additives commonly used in the art may be used. Compositions containing the active compound may be administered enterally or parenterally, with oral administration being preferred. The content of active compound in the composition is from about 0.5 to 100%, preferably from about 0.5 to about 20%, based on the total weight of the composition.

DETAILED DESCRIPTION OF THE INVENTION

Experiment 1:

Effect on Calcium Removal in SR Bladders Prepared from Heart Muscle and Solid Skeletal Muscle

Inhibitory effects of a given compound on phospholambane can be evidenced by measuring the effect of the compound on calcium depletion in SR vesicles. · · · ······ · · · · · · · · · · · · · · · · · · · · · · · · bladders prepared from solid skeletal muscles (hip and thigh muscles). Both types of SR bladders contain Ca ²⁺ -ATPase, but solid skeletal muscle bladders do not contain phospholambane (Hoh JFY, "Muscle fiber types and function", Current Opinion in Rheumatology, 4: 801-808, 1992). An increase in calcium removal in SR bladders prepared from cardiac tissue but not in SR bladders prepared from solid skeletal muscle indicates that the compound affects the inhibitory effect of phospholambane on SR Ca -ATP-as and thus acts as a phospholambane inhibitor. Since phospholambane limits both the rate, relaxation and contraction of the mammal's heart through its inhibitory effect on cardiac SR Ca -ATPase, compounds affecting these effects are potentially useful for the treatment of heart failure.

Method

Guinea pigs (10-12) were decapitated. Their hearts or lumbar muscles were resected, washed with ice-cold 0.9% NaCl and cut into pieces in a buffer containing 20 mM Tris-maleat, 0.3 M sucrose, pH 7.0. The tissue pieces were then homogenized with Polytron followed by Potter (10 shocks), the homogenates were centrifuged at 100 g for 15 min at 4 ° C. The supernatant was collected and the pellet was resuspended in 5 ml of buffer (20 mM Tris-maleat, 0.3 M sucrose, pH 7.0) and centrifuged again at 1000 g for 10 min at 4 ° C. The supernatant obtained was combined with the previously collected supernatant and centrifuged once more at 10,000 g for 20 min at 4 ° C. The resulting supernatant was filtered into a flask equipped with a magnetic stirrer. KCl was added to the filtered supernatant to a final concentration of 0.6 M (at 4 ° C). The resulting solution was centrifuged at 100,000 g for 60 min at 4 ° C. The pellet was suspended in 5 ml of a buffer containing 20 mM Tris-maleat, 0.3 M sucrose, pH 7.0, and centrifuged at 100,000 g for 60 min at 4 ° C. The obtained pellet was suspended in 5 ml of a buffer containing 20 mM Tris-maleat, 0.3 M sucrose, 0.1 M KCl, pH 7.0, and stored at -80 ° C until use. Protein concentration was also measured to standardize separately prepared vesicle preparations.

In the calcium removal rate assay, a fluorescent indicator, fluo-3, was used to detect a decrease in extravesicular Ca ²⁺ concentration when SR Ca ²⁺ ATPase transfers Ca ²⁺ from the extravesicular space to SR vesicles.

The SR-vesicles obtained above (50 µg protein / ml) were preincubated with or without test compound at 37 ° C for 5 min in assay buffer containing 40 mM imidazole, 95 mM KCl, 5 mM NaN ₃ , 5 mM MgCl ₂ , 0. , 5 mM EGTA, 5 mM potassium oxalate, 2 μΜ ruthenium red, 5 μΜ fluo-3, pH 7.0. Free calcium was adjusted to 0.1 μΜ or 0.04 μΜ with CaCl ₂ . The reaction was initiated by the addition of ATP (5 mM). The resulting reaction volume was

1.5 ml. Fluorescence in the reaction mixture was measured after 3 min using the excitation and emission wavelengths 510 nm and 530 nm.

Results

Giant. IA and 1B illustrate the effect of the compound of Example 1c (50 and 100 μΜ) on the rate of Ca removal in the SR of the bladders of the heart (A) and solid skeletal muscle (B). It can be seen that the compound of the invention accelerated calcium removal in heart SR bladders, but did not alter calcium removal in SR bladders prepared from solid skeletal muscle.

Table 1 shows the effects of various other phospholambane inhibitors 24 * of formula (I) or (II) on the rate of Ca removal in SR of the bladders of heart (A) and solid skeletal muscle (B). The experiments were carried out at a free calcium concentration of 0.1 μΜ and 0.04 μΜ.

TABLE 1: Stimulation (%) of Ca ²⁺ removal in bladder preparations obtained from ventricular myocardium (A) and solid skeletal muscle (B) from guinea pig heart.

• ·

The compound of Example no. (100 μΜ) Stimulation (%) Ca ²⁺ withdrawal AND (B) 3c ** 51 0 2c 26 -1 7c 5 -17 8g * ¹⁸ 0 11b 28 nd 12 32 nd 13d *** 23 nd 14c * 18 nd 18e 13 nd 21 11 nd 23 ** * * 20 May nd

* 10 μΜ, ** 20 μΜ, *** 50 μΜ, **** 5 μΜ nd = not determined

Experiment 2

Effects of derivatives on left ventricular pressure

Method

Guinea pigs of any sex weighing 300-400 g were used for the study. After the guinea pig was killed by a blow to the skull and decapitation, the heart was rapidly resected. Then, the heart was rinsed with cold oxygenated wash buffer. A cannula was inserted into the aorta and secured by ligature. Retrograde washes began as soon as the heart was placed in a thermostatically controlled humid chamber in a Langendorf device. Modified Tyrode's solution (37 ° C), balanced in a thermostatically controlled flask with carbogen (95% O ₂ and 5% CO ₂ ), was used as the wash buffer. Composition • 4 • · ·

Tyrode's solution was (in mM): NaCl 135; MgCl ₂ x 6H ₂ O 1; KCl 5; CaCl ₂ x 2H ₂ O 2; NaHCO ₃ 15; Na ₂ HPO ₄ x 2H ₂ O 1; glucose 10; pH 7.3 - 7.4. The experiments were carried out under constant pressure conditions (50 mm Hg).

After a short pre-stabilization (10 min), a latex balloon (size 4) was carefully inserted into the left ventricle through the left pulmonary vein and left atrium. The latex balloon was connected to a stainless steel cannula connected to a pressure sensor. The latex balloon, cannula, and pressure sensor chamber were filled with ethylene glycol / water (1: 1) to prevent any air bubbles. The isovolumetric pressure of the left ventricular was recorded through a pressure sensor. At the start of the experiment, the balloon volume was adjusted to obtain a diastolic pressure of 5 mmHg on average. Before starting the experiment, the heart was allowed to stabilize for an additional 30 to 50 minutes with vehicle (0.1% DMSO) in wash buffer.

After a 15 min baseline, different concentrations of test compound were added to the wash buffer at 15 min intervals. A concentration range of 0.3 to 30 μΜ was tested. Vehicle concentration (0.1% DMSO) was kept constant throughout the experiment.

Results

EC ₅₀ values and maximum effects (% change from baseline) for the various compounds of the invention on left ventricular systolic pressure are shown in Table 2.

• it

• · · · · · · · ·

TABLE 2: EC ₅₀ values and maximum effects (% change from baseline) on left ventricular systolic pressure

Compound ec ₅₀ maximum effect from example no. (μΜ) (%) lc 9 +52 at 30 μΜ 3c 4 +63 at 10 μΜ 5c . 10 + 14 at 30 μΜ 6c 0.5 +25 at 10 μΜ 7c 2.5 +29 at 10 μΜ 8g 2 +64 at 10 μΜ 9d 5 +50 at 30 μΜ 12 5 +22 at 10 μΜ 13d 10 +48 at 30 μΜ 14c 1.5 +25 at 10 μΜ 15c 3 +37 at 10 μΜ 16c 10 +57 at 30 μΜ 18e 10 +35 at 30 μΜ 19e 6 +39 at 30 μΜ

Experiment 3:

Effects on developing heart weakness in Langendorff guinea pigs isolated heart

Method

Guinea pigs of any sex weighing 300-400 g were used for the study. After the guinea pig was killed by a blow to the skull and decapitation, the heart was rapidly resected. Then, the heart was rinsed with cold oxygenated wash buffer. A cannula was inserted into the aorta and secured by ligature. Retrograde washes began as soon as the heart was placed in a thermostatically controlled humid chamber in a Langendorff apparatus. Modified Tyrode's solution (37 ° C), equilibrated in a thermostatic thermostat, was used as the wash buffer. A controlled oxidant flask with carbogen (95% 0) was used. ₂ and 5% CO ₂ ). The composition of Tyrode's solution was (in mM): NaCl 135; MgCl ₂ x 6H ₂ O 1; KC1 5; CaCl ₂ x 2H ₂ O 2; NaHCO ₃ 15; Na ₂ HPO ₄ x 2H ₂ O 1; glucose 10; pH 7.3 - 7.4. The experiments were performed under constant pressure conditions (50 mm Hg). After a short pre-stabilization (10 min), a latex balloon connected through a stainless steel cannula to a pressure sensor was carefully inserted into the left ventricle through the left pulmonary vein and left atrium. The latex balloon, cannula, and pressure sensor chamber were filled with ethylene glycol / water (1: 1) to prevent any air bubbles. The isovolumetric pressure of the left ventricular was recorded through a pressure sensor. At the start of the experiment, the balloon volume was adjusted to obtain a final diastolic pressure of 5 mmHg on average. Prior to the start of the experiment, the spontaneously beating heart was allowed to stabilize for an additional 30 to 50 minutes with vehicle (0.1% DMSO) in wash buffer.

After 15 min of baseline recording, the compound of Example 8g (10 μ přid) was added to the wash buffer. The heart was exposed 15 minutes later to global ischemia for 8 minutes, followed by reperfusion. This procedure was then repeated twice at 35 minute intervals. Another series of experiments was performed with vehicle instead of the compound of Example 8g. Vehicle concentration (0.1% DMSO) was kept constant during the experiments. The baseline was the average of the two minute recordings obtained just before the compound of Example 8g or the vehicle was added to the wash solution. Preischemia values were the average of two minute readings obtained just before each ischemic period, and reperfusion values were the average of two minute readings obtained at 8 min during each reperfusion period.

The results are shown in Figures 2A and B. Figure 2A shows the development of myocardial paralysis and subsequent decrease in left ventricular systolic pressure in the control group. Figure 2B shows that the phospholamban inhibitor of Example 8g completely inhibits the development of myocardial paralysis. Data are ± SEM of 2-3 experiments.

EXAMPLES

Example 1.

• · · ·

Preparation of 3-benzyl-5,7-bis [(1H-tetrazol-5-yl) methoxy] -4-methyl-2H-1-benzopyran-2-one

a) 3-Benzyl-5,7-dihydroxy-4-methyl-2 H -1-benzopyran-2-one

A solution of floroglucine dihydrate (20 g) and ethyl 2-benzylacetoacetate (27.5 mL) in ethanol (320 mL) was treated with dry HCl at 0 ° C for 5 hours and the solution was kept at this temperature overnight. The yellow solution was concentrated and triturated with water, the solids were filtered, washed with water and dried. The obtained hydrate was evaporated to dryness three times from toluene, triturated with petroleum ether (b.p. 40-60 ° C) and filtered. Yield: 33.4 g (96%). Mp 258-260 ° C.

1 H-NMR (DMSO-d ₆ , 400MHz): 2.525 / s, 3H, CH ₃ ), 3.887 (s, 2H, CH ₂ Ph), 6.171 (d, 1H, J = 2.4 Hz), 6.274 (d, 1H, J = 2.4Hz), 7.167-7.279 (m, 5H, Ph), 10.2 (s, 1H, OH), 10.47 (s, 1H, OH).

b) 3-Benzyl-5,7-bis (cyanomethoxy) -4-methyl-2 H -1-benzopyran-2-one

Chloroacetonitrile (6.86 g), potassium carbonate (23.9 g) and 12.2 g of the product of Example 1a were stirred in 120 mL of DMF at 100 ° C under nitrogen for 2 hours. The reaction mixture was cooled and poured into ice water. The solids • 4 • 4 were filtered and washed with water. Yield 13.8 g (88%). Melting point 147-154 ° C.

1 H-NMR (DMSO-d ₆ , 400MHz): 2.525 (s, 3H, CH ₃ ), 3.969 (s, 2H, CH ₂ Ph), 5.307 (s, 2H, OCH 2 CN), 5.314 (s, 2H, OCH 2 CN), 6.814 (d, 1H, J =

2.5 Hz), 6.940 (d, 1H, J = 2.5 Hz), 7.18-7.292 (m, 5H, Ph).

c) 3-benzyl-5,7-bis [(1H-tetrazol-5-yl) methoxy] -4-methyl-2 H -1-benzopyran-2-one

The product of Example 1b (1 g), sodium azide (0.42 g) and ammonium chloride (0.34 g) were stirred in DMF (5 mL) under nitrogen at 100 ° C for 5 hours. The reaction mixture was allowed to cool and then poured into ice water. The pH of the solution was adjusted to 10-11 and then the solution was either extracted once with ethyl acetate or filtered through Celite. The solution was acidified to pH 2 with hydrochloric acid, stored at 5 ° C and filtered. Yield 0.96 g (81%). Melting point 229-233 ° C.

1 H-NMR (DMSO-d ₆ , 400MHz): 2.468 (s, 3H, CH ₃ ), 3.937 (s, 2H, CH ₂ Ph), 5.596 (s, 2H, OCH ₂ Tet), 5.602 (s, 2H, _OCH2 Tet), 6.832 (d, 1H, J =

2.4 Hz), 6.851 (d, 1H, J = 2.4 Hz), 7.171-7.283 (m, 5H, Ph).

Example 2

Preparation of 7,8,9,10-tetrahydro-1,3-bis [(1H-tetrazol-5-yl) methoxy] -7-phenyl-6H-dibenzo [b, d] pyran-6-one · ·

(a) 7,8,9,10-tetrahydro-1,3-dihydroxy-7-phenyl-6H-dibenzo [b, d] pyran-6-one

A solution of floroglucin (0.7 g) and 2-ethoxycarbonyl-3-phenylcyclohexanone (1.5 g) in ethanol was treated with dry HCl as described in Example 1a. The product was first recrystallized from ethanol-water (1: 1) and then triturated with ether. Yield 0.61 g.

1 H-NMR (DMSO-d ₆ , 400MHz): 1.38-1.52 (m, 1H), 1.57-1.66 (m, 1H), 1.69-1.78 (m, 1H) ), 1.86-1.96 (m, 1H), 2.9-3.02 (m, 1H), 3.3-3.4 (m, 1H), 4.050 (b, 1H), 6.157 ( d, 1H, J = 2.4 Hz), 6.297 (d, 1H, J = 2.4 Hz), 7.076 7.265 (m, 5H), 10.153 (s, 1H), 10.456 (s, 1H).

b) 7,8,9,10-tetrahydro-1,3-bis (cyanomethoxy) -7-phenyl-6H-dibenzo [b, d] pyran-6-one

The product of Example 2a (0.5 g) was treated with chloroacetonitrile (0.25 g) and potassium carbonate (1.12 g) in DMF (5 mL) as described in Example 1b. Yield 0.6 g.

1 H-NMR (DMSO-d ₆ , 400MHz): 1.38-1.58 (m, 1H), 1.6-1.7 (m, 1H), 1.7-1.76 (m, 1H) 1.89 - 1.99 (m, 1H), 2.9 - 3.03 (m, 1H), 3.2 - 3.28 (m, 1H), 4.111 (b, 1H), 5, 314 (s, 2H), 5.349 (s, 2H), 6.840 (d, 1H, J = 2.5Hz), 6.925 (d, 1H, J = 2.5Hz), 7.108-7.274 (m, 5H) .

• · · «· · · · · · · · · · ·

c) 7,8,9,10-tetrahydro-1,3-bis [(1H-tetrazol-5-yl) methoxy] -7-phenyl-6H-

The product of Example 2b (0.6 g) was treated with sodium azide (0.2 g) and ammonium chloride (0.17 g) in DMF (5 mL), as in Example 1c. The product was recrystallized from a mixture of DMF, ethanol and water (approximately 1: 2: 3). Yield 0.41 g. M.p .: 153-154 ° C.

1 H-NMR (DMSO-d ₆ , 400MHz): 1.38-1.5 (m, 1H), 1.5-1.6 (m, 1H), 1.69-1.76 (m, 1H) ), 1.87-1.96 (m, 1H), 2.9-3.05 (m, 1H), 3.2-3.3 (m, 1H), 4.094 (b, 1H), 5.602 ( s, 2H), 5.643 (s, 2H), 6.832 (d, 1H, J = 2.3 Hz), 6.851 (d, 1H, J = 2.3 Hz), 7.089-7.212 (m, 5H).

Example 3

Preparation of 3-benzyl-5,7-bis [(2,5-dihydro-5-oxo-4H-1,2,4-oxadiazol-3-yl) methoxy] -4-methyl-2H-1-benzopyran-2 -onu

a) 3-Benzyl-5,7-bis [(hydroxyamidino) methoxy] -4-methyl-2 H -1-benzopyran-2-one

Triethylamine (1.94 mL) was added to a suspension of hydroxylamine hydrochloride (0.97 g) in DMSO (2 mL) and the resulting mixture was stirred at room temperature for 1 h. ♦ · 4 · 4 · 4 ·

4 9 9 9

9 4 44 4 * 4 4 minutes. The crystals were filtered and washed with THF. The filtrate was concentrated and the product of Example 1b (0.5 g) was added. This solution was maintained at 75 ° C overnight. The reaction mixture was treated with ice water, the pH was adjusted to 11 and the solids were filtered off, washed with water and dried. Yield 0.5 g. M.p. 155-160 ° C.

1 H-NMR (DMSO-d ₆ , 400MHz): 2.56 (s, 3H, CH ₃ ), 3.938 (s, 2H), 4.466 (s, 2H), 4.466 (s, 2H), 5.5675 ( s, H, NH ₂ ), 5.709 (s, 2H, NH ₂ ), 6.658 (d, 1H, J = 2.4 Hz), 6.692 (d, 1H, J = 2.4 Hz), 7.168-7.284 ( m, 5H, Ph), 9.346 (s, 1H, OH), 9.362 (s, 1H, OH).

b) 3-Benzyl-5,7-bis [(ethoxycarbonyloxyamidino) methoxy] -4-methyl-2H-1-benzopyran-2-one

o-Ethyl chloroformate (0.45 mL) was added to a solution of the product of Example 3a (1 g) and pyridine (0.38 mL) in DMF (5 mL) at 0 ° C. The reaction mixture was maintained at this temperature for an additional 30 minutes and then ice water was added. The solids were filtered and washed with water. The yield was 1.63 g. M.p. 87-92 ° C.

1 H-NMR (DMSO-d ₆ , 400mHz): 1.215-1.256 (m, 6H), 2.553 (s, 3H), 3.947 (s, 2H), 4.140-4.198 (m, 4H), 4.566 (s, 2H) ), 4.599 (s, 2H), 6.688 (d, 1H, J = 2.4 Hz), 6.718 (d, 1H, J = 2.4 Hz), 6.792 (b, 2H, NH ₂ ), 7.171-7.285 (m, 5 H).

c) 3-benzyl-5,7-bis [(2,5-dihydro-5-oxo-4H-1,2,4-oxadiazol-3-yl) methoxy] -4-methyl-2H-1-benzopyran- 2-on • 0 09 40 0999 90 '44 · 04 · 900,000

0 9 4 4091 0 44 9

The product of the previous example (1.5 g) and DBU (0.8 mL) in DMF (5 mL) was stirred at room temperature overnight. The reaction mixture was treated with ice and acidified. The solids were filtered and washed with water. The solid obtained was taken up in 0.1 N sodium hydroxide solution, treated with activated carbon and finally acidified. The yield was 0.64 g. Melting point: 130136 ° C.

1 H-NMR (DMSO-d ₆ , 400MHz): 2.524 (s, 3H), 3.954 (s, 2H), 5.187 (s, 2H), 5.215 (s, 2H), 6.748 (d, 1H, J = 2) 4 Hz), 6.834 (d, 1H, J = 2.4 Hz), 7.158-7.289 (m, 5H), 12.8 (b, 2H).

Example 4

Preparation of 7,8,9,10-Tetrahydro-bis [(1H-tetrazol-5-yl) methoxy] -1,3-dihydroxy-6H-dibenzo [b, d] pyran-6-one

(a) 7,8,9,10-Tetrahydro-1,3-dihydroxy-6H-dibenzo [b, d] pyran-6-one

Floroglucinol (1 g) and ethyl 2-oxocyclohexanecarboxylate (1.32 g) were stirred in 75% sulfuric acid (10 mL) overnight, poured into ice water and filtered. Yield: 1.55 g.

9 9 9 9 9

9 9 9

9991 • 9 9,999

97 ► 9 9 9

I

9 99

1 H-NMR (DMSO-d ₆ , 400 MHz): 1.65 (b, 4H), 2.355 (b, 2H), 3.037 (b, 2H), 6.138 (d, 1H, J = 2.4 Hz), 6.245 (d, 1H, J = 2.4Hz), 10.069 (b, 1H, OH), 10.322 (s, 1H, OH).

b) 7,8,9,10-Tetrahydro-bis (cyanomethoxy) -1,3-dihydroxy-6H-dibenzo [b, d] pyran-6-one

The product of the previous example (0.5 g), chloroacetonitrile (0.34 g) and potassium carbonate (1.5 g) in DMF (5 mL) were reacted as in Example 1b. Yield: 0.44 g.

1 H-NMR (DMSO-d 6, 400 MHz): 1.68 (b, 4H), 2.41 (b, 2H), 3.00 (b, 2H), 5.297 (s, 2H), 5.309 (s, 2H) 6.797 (d, 1H, J = 2.4 Hz); 6.899 (d, 1H, J = 2.4 Hz).

c) 7,8,9,10-Tetrahydro-bis [(1H-tetrazol-5-yl) methoxy] -1,3-dihydroxy-6H-dibenzo [b, d] pyran-6-one

The product of the previous example (0.4 g) was treated with sodium azide (0.18 g) and ammonium chloride (0.14 g) in DMF (2.5 mL) as in Example 1c. The product was recrystallized from ethanol-DMF (1: 1). The yield was 0.17 g. Mp 283-286 ° C.

1 H-NMR (DMSO-d 6, 400 MHz): 1.626 (b, 4H), 2.393 (b, 2H), 2.971 (b, 2H), 5.583 (s, 2H), 5.599 (s, 2H), 6.811 (s, 2H) ).

<·· ♦

Example 5

Preparation of 5,7-bis [(1H-tetrazol-5-yl) methoxy] -4-phenyl-2H-1-benzopyran-2-one

a) 5,7-Dihydroxy-4-phenyl-2H-1-benzopyran-2-one;

A solution of floroglucin (2.00 g) and ethyl benzoylacetate (3.05 g) in ethanol (30 mL) was treated with dry HCl as described in Example 1 a. The product was recrystallized from ethanol-water (1: 1). Yield 3.0 g. (75%).

¹ H-NMR (DMSO-d ₆ , 400 MHz): 5.739 (s, 1H, CH = C), 6.15 (d, 1H, J =

2.3 Hz), 6.263 (d, 1H, J = 2.3 Hz), 7.305-7.381 (m, 5H, Ph), 10.084 (s, 1H, OH), 10.368 (s, 1H, OH).

b) 5,7-bis (cyanomethoxy) -4-phenyl-2H-1-benzopyran-2-one

The product of the previous example (1.00 g) was treated with chloroacetonitrile (0.62 g) and potassium carbonate (2.72 g) in DMF (5 mL) as described in Example 1B. The reaction mixture was poured into ice water and the mixture was extracted with ethyl acetate. The ethyl acetate was washed with 1M NaOH, dried over sodium sulfate and evaporated. The product was recrystallized from isopropanol. Yield 0.41 g (31%).

1 H-NMR (DMSO-d 6, 400 MHz): 4.845 (s, 2H, OCH ₂ CN), 5.344 (s, 2H, OCH ₂ CN), 6.086 (s, 1H, CH = C), 6.770 (d, 1H, J) = 2.4 Hz), 7.040 (d, 1H, J =

2.4 Hz), 7.320-7.443 (m, 5H, Ph).

c) 5,7-Bis [(1H-tetrazol-5-yl) methoxy] -4-phenyl-2H-1-benzopyran-2-one

ΦΦΦ φ φ φ φ • • • φ φ φ φ φ φ φ φ φ

ν = ν

The product of the previous example (0.40 g) was treated with sodium azide (0.16 g) and ammonium chloride (0.14 g) in DMF (2 mL) at 100 ° C for 2 hours. The product was isolated as described in Example 1C. Yield: 0.40 g (79%). Melting point 222-224 ° C.

¹ H-NMR (DMSO-d ₆ , 400 MHz): 5.148 (s, 2H, OCH ₂ Tet), 5.649 (s, 2H, OCH ₂ Tet), 5.968 (s, 1H, CH = C), 6.811 (d 1 H, J = 2.3 Hz), 6.962 (d, 1H, J = 2.3 Hz), 6.994-7.185 (m, 5H, Ph).

Example 6

Preparation of 7,8,9,10-Tetrahydro-1,3-bis [(1H-tetrazol-5-yl) methoxy] -8-phenyl-6H-dibenzo [b, d] pyran-6-one

(a) 7,8,9,10-Tetrahydro-1,3-dihydroxy-8-phenyl-6H-dibenzo [b, d] pyran-6-one

A solution of floroglucin (1.56 g) and ethyl 2-oxo-5-phenylcyclohexanecarboxylate (2.52 g) in ethanol (25 mL) was treated with dry HCl as described in Example 1a. The precipitate was filtered off and washed with water and EtOH. The yield was 1.0 g (32%).

1 H-NMR (DMSO-d ₆ , 400 MHz): 1.72-1.82 (m, 1H), 2.01 (b, 1H), 2.3172.387 (m, 1H), 2.707-2.763 (m, 1H), 2.830 (b, 1H), 3.031 (b, 1H), 3.35 and 3.40 to · ··············· · · · · 9 · 9 · · · • · • · · · · · toto toto toto toto toto toto toto toto toto toto toto toto toto (b, 1H), 6.174 (d, 1H, J = 2.3 Hz), 6.277 (d, 1H, J = 2.3 Hz), 7.200-7.350 (m, 5H, Ph), 10.131 (s, 1H) (OH), 10.401 (s, 1H, OH).

b) 7,8,9,10-Tetrahydro-1,3-bis (cyanomethoxy) -8-phenyl-6H-dibenzo [b, d] pyran-

The product of the previous example (1.0 g) was treated with chloroacetonitrile (0.57 g) and potassium carbonate (1.0 g) in DMF (5 mL) as described in Example 1b. The DMF was evaporated and the residue was dissolved in EtOAc. The ethyl acetate was washed with 1 M NaOH, dried with sodium sulfate and evaporated. The product was recrystallized from acetone-isopropanol (1: 3). The yield was 0.50 g (40%).

1 H-NMR (DMSO-d ₆ , 300 MHz): 1.75-1.88 (m, 1H), 2.05 (b, 1H), 2.382.48 (m, 1H), 2.77-2 85 (m, 1H), 2.90 (b, 1H), 3.07 (b, 1H), 3.22 and 3.28 (b, 1H), 5.316 (s, 2H, OCH ₂ CN), 5.331 (s, 2H, _OCH2 CN), 6.829 (d, 1H, J = 2.5 Hz), 6.939 (d, 1H, J = 2.5 Hz), 7.210 to 7.380 (m, 5H, Ph).

c) 7,8,9,10-Tetrahydro-1,3-bis [(1H-tetrazol-5-yl) methoxy] -8-phenyl-6H-dibenzo [b, d] pyran-6-one

The product of the previous example (0.30 g) was treated with sodium azide (0.10 g) and ammonium chloride (0.09 g). in DMF (12 mL) at 100 ° C for 3.5 hours. The product was isolated in the same manner as in Example 1c. The yield was 0.30 g (82%). Mp 235-245 ° C.

* 4 ···· «4 ·· φ 4« 4 4 · 4 · 4 »4 · · · · 44 Φ 4 4 4

4,444 «4444 4

449 9 9 9 9 9 9 9

4444 44 444 44 44 ¹ H-NMR (DMSO-d ₆ , 400 MHz): 1.70-1.80 (m, 1H), 1.96 (b, 1H), 2.382.446 (m, 1H), 2.836 ( m, 2H), 3.052 (b, 1H), 3.252 and 3.301 (b, 1H), 5.604 (s,

2H, _OCH2 CN), 5.632 (s, 2H, _OCH2 CN), 6.827 (d, 1H, J = 2.5 Hz), 6.858 (d,

1H, J = 2.5 Hz), 7.209-7.351 (m, 5H, Ph).

Example 7

Preparation of 5,7-bis [(1H-tetrazol-5-yl) methoxy] -4-methyl-3- (2-phenylethyl) -2H-1-benzopyran-2-one

(a) 5,7-Dihydroxy-4-methyl-3- (2-phenylethyl) -2H-1-benzopyran-2-one

A solution of floroglucin (0.87 g) and ethyl 2- (2-phenylethyl) -acetoacetate (1.62 g) in ethanol (30 ml) was treated with dry HCl as described in Example 1a. The yield was 1.77 g (87%). Mp 248-252 ° C.

1 H-NMR (DMSO-d 6, 300 MHz): 2.413 (s, 3H, CH ₃ ), 2.652-2.782 (m, 4H, CH ₂ CH ₂ ), 6.151 (d, 1H, J = 2.4 Hz), 6.256 (d, 1H, J = 2.4Hz), 7.1837.304 (m, 5H, Ph), 10.137 (s, 1H, OH), 10.369 (s, 1H, OH).

b) 5,7-Bis (cyanomethoxy) -4-methyl-3- (2-phenylethyl) -2H-1-benzopyran-2-one

The product of the previous example (0.90 g) was treated with chloroacetonitrile (0.48 g) and potassium carbonate (2.1 g) in DMF (5 mL) at 100 ° C for 0.5 h. The product was isolated as described in Example 1b. The yield was 1.00 g (88%). M.p. 179-183 ° C.

• · • ·

1 H-NMR (DMSO-d ₆ , 300 MHz): 2.384 (s, 3H, CH ₃ ), 2.699-2.754 (m, 2H, CH ₂ CH ₂ ), 2.805-2.841 (m, 2H, CH ₂ CH ₂₎ ), 5.302 (s, 4H, OCH ₂ CN), 6.790 (d, 1H, J = 2.5 Hz), 6.909 (d, 1H, J = 2.5 Hz), 7.190-7.307 (m, 5H, Ph ).

c) 5,7-Bis [(1H-tetrazol-5-yl) methoxy] -4-methyl-3- (2-phenylethyl) -2H-1-benzopyran-2-one

The product of the previous example (0.40 g) was treated with sodium azide (0.15 g) and ammonium chloride (0.12 g) in DMF (2 mL) at 100 ° C for 2.5 hours. The product was isolated as described in Example 1c. The yield was 0.385 g (78%). Mp 248-250 ° C.

1 H-NMR (DMSO-d 6, 400 MHz): 2.368 (s, 3H, CH ₃ ), 2.688-2.707 (m, 2H, CH ₂ CH ₂ ), 2.783-2.822 (m, 2H, CH ₂ CH ₂ ), 5.593 (s, 2H, OCH ₂ Tet), 5.604 (s, 2H, OCH ₂ Tet), 6.819 (d, 1H, J = 2.3 Hz), 6.834 (d, 1H, J = 2.3 Hz), 7.161 7.291 (m, 5H, Ph).

Example 8

Preparation of 5,7-bis [(1H-tetrazol-5-yl) methoxy] -1,3-dibenzyl-4-methyl-2 (1H) -quinoline

(a) 2-Benzyl-3-oxobutanoic acid 3,5-dimethoxyanilide

3,5-Dimethoxyaniline (5 g) was added in portions to preheated (160 ° C) ethyl 2-benzylacetoacetate (15 mL) under nitrogen and was maintained at this temperature. ···· · · ··· · ····· ···· · · · · · · ·

1Λ ··· ·· · ····

Temperature at 60 minutes. The cooled solution was diluted with heptane ethyl ether and filtered. The yield was 5.2 g (49%).

1 H-NMR (DMSO-d ₆ , 300 MHz): 2.183 (s, 3H), 3.069 (d, 2H, J = 7.2 Hz), 3.923 (t, 1H, J = 7.2 Hz), 6.616 (dd, 1H, J = 2.3 Hz), 6.765 (d, 2H, J = 2.3 Hz), 7.13-7.3 (m, 5H), 10.123 (s, 1H).

b) 3-Benzyl-5,7-dimethoxy-4-methyl-2- (1H) -quinoline

The product of the previous example (1.2 g) was added to preheated (85 ° C) methanesulfonic acid (3.5 mL) and maintained at this temperature for 15 minutes. The solution was allowed to cool and then treated with ice water. The product was filtered, washed with sodium bicarbonate and water. Yield

1.0 g (95%).

1 H-NMR (100 MHz): 2.486 (s, 3H), 3.785 (s, 3H), 3.808 (s, 3H), 3.985 (s, 2H), 6.315 (d, 1H, J = 2.4 Hz), 6.472 (d, 1H, J = 2.4Hz), 7.1-7.3 (m, 5H),

II, 52 (s, 1 H).

c) 3-Benzyl-5,7-dihydroxy-4-methyl-2 (1H) -quinolinone

The product of the previous example (1 g) was refluxed under nitrogen in pyridine hydrochloride (5 g) for 20 minutes. The reaction mixture was treated with water and the product was filtered. The yield was 0.9 g (100%). Melting point: 307-312 ° C.

1 H-NMR (300 MHz): 2.503 (s, 3H), 3.942 (s, 2H), 6.102 (d, 1H, J = 2.3 Hz), 6.187 (d, 1H, J = 2.3 Hz) 7.1-7.25 (m, 5H), 9.725 (s, 1H), 9.984 (s, 1H), 11.285 (s, 1H).

• ·

d) 1,3-Dibenzyl-5,7-dimethoxy-4-methyl-2 (1H) -quinolinone

The product of Example 8b (1 g), potassium tert-butoxide (0.62 g) and benzyl bromide (0.68 mL) were stirred in DMSO (10 mL) at 60 ° C for 4 hours. The reaction mixture was treated with water, extracted with toluene and evaporated. The product was triturated with ethyl ether and filtered. The yield was 0.5 g (39%).

1 H-NMR (400 MHz): 2.537 (s, 3H), 3.708 (s, 3H), 3.826 (s, 3H), 4.124 (s, 2H), 5.56 (b, 2H), 6.413-6.434 ( m, 2H), 7.154-7.332 (m, 10H).

e) 1,3-Dibenzyl-5,7-dihydroxy-4-methyl-2 (1H) -quinolinone

The product of the previous example (2 g) was treated with pyridine hydrochloride (10 g) as described in Example 8c. The product was extracted with ethyl acetate and evaporated. The yield was 1.4 g (75%).

1 H-NMR (400 MHz), 2.570 (s, 3H), 4.076 (s, 2H), 5.450 (b, 2H), 6.135 (d, 1H, J = 2.2 Hz), 6.199 (d, 1H) J = 2.2 Hz), 7.128-7.333 (m, 10H), 9.83 (b, 1H), 10.166 (s, 1H).

f) 5,7-Bis (cyanomethoxy) -1,3-dibenzyl-4-methyl-2 (1H) -quinolinone

N O

The product of the previous example (1.4 g) was treated with chloroacetonitrile (0.76 g) and K ₂ CO ₃ (2.5 g) in DMF (20 mL) as described in Example 1b. The yield was 1.5 g (89%).

1 H-NMR (400 MHz): 2.555 (s, 3H), 4.146 (s, 2H), 5.214 (s, 2H), 5.275 (s, 2H), 5.578 (s, 2H), 6.735 (s, 2H) 7.13-7.33 (m, 10H).

g) 5,7-Bis [(1H-tetrazol-5-yl) methoxy] -1,3-dibenzyl-4-methyl-2 (1H) -quinolinone

The product of the previous example (1.3 g) was treated with sodium azide (0.41 g) and ammonium chloride (0.34 g) as described in Example 1c. The yield was 0.69 g (45%).

¹ H-NMR (400 MHz): 2.471 (s, 3H), 4.113 (s, 2H), 5.477 (s, 2H), 5.55 (b, 2H), 5.574 (s, 2H), 6.670 (d) 1 H, J = 2.1 Hz), 6.775 (d, 1H, J = 2.1 Hz),

7.13-7.32 (m, 10H).

Example 9

Preparation of 5,7-bis [(1H-tetrazol-5-yl) methoxy] -3-benzyl-1,4-dimethyl-2 (1H) -quinolinone

(a) 3-Benzyl-5,7-dimethoxy-1,4-dimethyl-2 (1H) -quinolinone

The product of Example 8b (0.5 g), t-BuOK (0.2 g) and methyl iodide (0.4 mL) were stirred in DMSO (5 mL) at 35 ° C for two days. The reaction mixture was treated with water and extracted with toluene. The product was purified by chromatography on 0 0 0 0 0 0 0 0 0 0

0 · 00000 * ···

0 0 · 0 0 000 000 0

0-1 00 0 0000000 · ««? 00 0000 00 000 ·· 00 column using toluene-ethyl acetate-acetic acid 8: 2: 1 as eluent.

The yield was 0.24 g (46%).

1 H-NMR (300 MHz): 2.51 (s, 3H), 3.632 (s, 2H), 3.846 (s, 3H), 3.896 (s, 3H), 4.047 (s, 2H), 6.468 (d) 1 H, J = 2.3 Hz), 6.558 (d, 1H, J = 2.3 Hz),

7.1-7.26 (m, 5H);

b) 3-Benzyl-5,7-dihydroxy-1,4-dimethyl-2 (1H) -quinolinone

The product of the previous example (0.2 g) was treated with pyridine hydrochloride (2 g) as described in Example 8c, and the product was extracted with ethyl acetate. The yield was 0.16 g (89%).

¹ H-NMR (400 MHz): 2.567 (s, 3H), 3.515 (s, 3H), 4.005 (s, 2H), 6.244 (d, 1H, J = 2.3 Hz), 6.268 (d, 1H) J = 2.3 Hz), 7.08-7.25 (m, 5H), 9.879 (s, 1H), 10.113 (s, 1H).

c) 5,7-Bis (cyanomethoxy) -3-benzyl-1,4-dimethyl-2 (1H) -quinolinone

The product of the previous example (0.15 g), chloroacetonitrile (0.08 g) and K ₂ CO ₃ (0.28 g) were reacted in DMF (2 mL) as described in Example 1b. The yield was 0.16 g (84%).

1 H-NMR (400 MHz): 2.524 (s, 3H), 3.658 (s, 3H), 4.079 (s, 2H), 5.292 (s, 2H), 5.379 (s, 2H), 6.766 (d, 1H) J = 2.3 Hz), 6.855 (d, 1H, J = 2.3 Hz),

7.13-7.24 (m, 5H).

• 9 • 9 • 9

9 9 9 9 9 9 9 9 9 99 99

d) 5,7-bis [(1H-tetrazol-5-yl) methoxy] -3-benzyl-1,4-dimethyl-2 (1H) -quinolinone

The product of the previous example (0.15 g) was treated with NaN ₃ (57 mg) and NH ₄ Cl (47 mg) in DMF (2 mL) as described in Example 1c. The yield was 0.115 g. Melting point: 250-253 ° C.

1 H-NMR (400 MHz): 2.451 (s, 3H), 3.649 (s, 3H), 4.042 (s, 2H), 6.792 (d, 1H, J = 2.2 Hz), 6.833 (d, 1H) J = Hz) 7.1-7.25 (m, 5H).

Example 10

Preparation of 3-Benzyl-5,7-bis [(2-methyl-1H-tetrazol-5-yl) methoxy] -4-methyl-2H-1-benzopyran-2-one and the three isomers

0.07 mL of methyl iodide was added to a solution of 0.2 g of the product of Example 1c and 0.31 g of K ₂ CO ₃ in 2 mL of DMF, and the mixture was stirred at room temperature for 4 hours. The reaction mixture was poured into ice water and filtered. The yield was 0.2 g as a mixture of four regioisomers, m.p. 71-76 ° C.

1 H-NMR (DMSO-d ₆ ; 400 MHz): 2.47 (s, CH ₃ ), 2.48 (s, CH ₃ ), 3.93 (s, CH ₂ Ph) 4.11 (s, NCH ₃ ), 4.15 (s, NCH ₃ ), 4.38 (s, NCH ₃ ), 4.40 (s, NCH ₃ ), 5.51 (s, OCH ₂ ) 5.52 (s, OCH ₂ ), 5.62 (s, OCH ₂ ), 5.67 (s, OCH ₂ ), 6.84-6.91 (m, 2H), 7.16-7, 28 (m, 5H, Ph).

Example 11

Preparation of 3-benzyl-5,7-bis [1- (1H-tetrazol-5-yl) ethoxy] -4-methyl-2H-1-benzopyran-2-one, mixtures of stereoisomers

a) 3-Benzyl-5,7-bis [(1-cyano) ethoxy] -4-methyl-2 H -1-benzopyran-2-one

The product of Example 1a (1 g), 2-chloropropionitrile (0.7 g) and potassium carbonate (2 g) were heated in DMF (15 mL) under nitrogen at 110 ° C for sixty minutes. The mixture was treated with water, filtered and washed with 1 N NaOH and water. The yield was 1.2 g.

1 H-NMR (DMSO-d ₆ , 300 MHz): 1.74-1.78 (t + t, 6H, CH-CH ₃ ), 2.53 (s, 3H), 3.97 (s , 2H), 5.58 to 5.66 (m, 2H, CH-CH _3), 6.87 (m, 1H), 6.99 (d, (1H), 7.18-7.31 (m , 5H).

b) 3-Benzyl-5,7-bis [1- (1H-tetrazol-5-yl) ethoxy] -4-methyl-2H-1-benzopyran-2-one, mixture of stereoisomers

About O · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · ·

The product of the previous example (0.5 g), sodium azide (0.18 g), and ammonium chloride (0.15 g) were heated in DMF (7 mL) at 100 ° C for 90 minutes. The product was treated with water, extracted with ethyl acetate and evaporated. The yield was 0.57 g. Melting point 91-104 ° C.

1 H-NMR (DMSO-d ₆ , 300 MHz): 1.69-1.77 (m, 6H, CH-CH ₃ ), 2.54 (s, 3H), 3.94 (s, 2H) ), 6.10 to 6.17 ((m, 2H, CH-CH _3), 6.65 (dd, IH), 6.74 (dd, IH),

7.13-7.30 (m, 5H).

Example 12

Preparation of 5,7-bis (carboxymethoxy) -1,3-dibenzyl-4-methyl-2 (1H) -quinolinone

The product of Example 8f (0.2 g) was refluxed in a solution of concentrated hydrochloric acid (3 mL) and acetic acid (2 mL) for one hour. The product was filtered at 25 ° C. The yield was 0.14 g.

1 H-NMR (DMSO-d ₆ , 300 MHz): 2.63 (s, CH ₃ ), 4.14 (s, 2H, CH ₂ Ph), 4.66 (s, 2H, OCH ₂ COOH) 4.79 (s, 2H, OCH ₂ COOH), 5.53 (s, 2H, NCH ₂ Ph), 6.41 (d, 1H, J = 2.2 Hz), 6.45 (d, 1H J = 2.2 Hz), 7.13-7.34 (m, 10H, Ph).

Example 13

Preparation of 3-benzyl-5,7-bis [(1H-tetrazol-5-yl) methoxy] -1- (4-fluorobenzyl) -4-methyl-2 (1H) -quinolinone

a) 1-Benzyl-5,7-dimethoxy-3- (4-fluorobenzyl) -4-methyl-2 (1H) -quinolinone

• f • • «« to toto toto toto <<<<<<^ ^ ^ ^ ^ ^ ^ ^ ^ ^ ^ ^ ¹ · · · · · · «· toto ·· ·· ··

The product of Example 8b (2 g), potassium tert-butoxide (0.87 g) and 4-fluorobenzyl chloride (1.12 g) were heated in DMSO (20 mL) at 60 ° C for three hours, as in Example 8d. The yield was 1.28 g.

1 H-NMR (DMSO-d ₆ , 400 MHz): 2.53 (s, 3H), 3.73 (s, 3H), 3.83 (s, 3H),

5.55 (s, 2H), 6.43 (s, 2H), 7.12-7.2 (m, 5H), 7.26-7.28 (m, 4H).

b) 3-Benzyl-5,7-dihydroxy-1- (4-fluorobenzyl) -4-methyl-2 (1H) -quinolinone

The product of the previous example (1.25 g) was heated in pyridine hydrochloride (12.5 g) at about 225 ° C for 9 minutes. The yield was 1 g.

1 H-NMR (DMSO-d ₆ , 300 MHz): 2.56 (s, 3H), 4.07 (s, 2H), 5.4 (b, 2H), 6.13 (d, 1H, J) = 2.1 Hz), 6.20 (d, 1H, J = 2.1 Hz), 7.12-7.28 (m, 9H), 9.88 (s, 1H), 10.22 (s , 1H).

c) 3-Benzyl-5,7-bis (cyanomethoxy) -1- (4-fluorobenzyl) -4-methyl-2 (1H) -quinolinone

The product of the previous example (1 g), ClCH ₂ CN (0.43 g) and K ₂ CO ₃ (1.42 g) were heated in DMF (8 mL) at 120 ° C for one hour. The yield was 0.94 g.

»4,444 • 4 ·

4 4

4 4 4 4 4

4 · 44 4 4 4 4 'η-NMR (DMSO-d _6, 300 MHz): 2.55 (s, 3H), 4.14 (s, 2H), 5.52 (s, 2H), 5, 28 (s, 2H), 5.57 (s, 2H), 6.74 (s, 2H, ArH), 7.1-7.3 (m, 9H).

d) 3-Benzyl-5,7-bis [(1H-tetrazol-5-yl) methoxy] -1- (4-fluorobenzyl) -4-methyl-2 (1H) -quinolinone

The product of the previous example (0.5 g), sodium azide (0.147 g) and ammonium chloride (0.12 g) were heated in DMF (5 mL) at 120 ° C for 90 minutes. The product was triturated with acetonitrile. Yield 0.28 g. M.p .: 126-132 ° C.

1 H-NMR (DMSO-d ₆ , 300 MHz): 2.48 (s, 3H), 4.11 (s, 2H), 5.51 (s, 2H), 5.55 (s, 2H), 5.58 (s, 2H), 6.67 (d, 1H, J = 2.1Hz), 6.78 (d, 1H, J = 2.1Hz).

Example 14

Preparation of 5,7-bis [(1H-tetrazol-5-yl) methoxy] -3- (4-chlorobenzyl) -4-methyl-2H-1-benzopyran-2-one

a) 3- (4-Chlorobenzyl) -5,7-dihydroxy-4-methyl-2 H -1-benzopyran-2-one

A solution of florglucin (1.57 g) and ethyl 2- (4-chlorobenzyl) acetoacetate (3.18 g) in ethanol (25 mL) was treated with dry HCl at 0 ° C for 1.5 hours and the solution was stored at at this temperature overnight. The solvent was evaporated and • · ··

the precipitate was triturated with water. The yield was 3.87 g (98%). Melting point 270-278 ° C.

1 H-NMR (DMSO-d ₆ , 300 MHz): 2.52 (s, 3H, CH ₃ ), 3.87 (s, 2H, CH ₂ ), 6.17 (d, 1H, J = 2, 4 Hz), 6.28 (d, 1H, J = 2.4 Hz), 7.18-7.34 (tn, 4H, Ph), 10.21 (s, 1H, OH), 10.48 ( s, 1H, OH).

b) 5,7-Bis (cyanomethoxy) -3- (4-chlorobenzyl) -4-methyl-2 H -1-benzopyran-2-one

Cl

The product of the previous example (1.00 g), chloroacetonitrile (0.50 g) and potassium carbonate (2.18 g) were heated in DMF (5 mL) at 100 ° C for 30 minutes. The product was isolated as described in Example 1b. The yield was 0.90 g (72%).

1 H-NMR (DMSO-d ₆ , 300 MHz): 2.52 (s, 3H, CH ₃ ), 3.95 (s, 2H, CH ₂ ), 5.308 (s, 2H, OCH ₂ CN), 5.312 (s, 2H, OCH ₂ CN), 6.81 (d, 1H, J = 2.5 Hz),

6.94 (d, 1H, J = 2.5 Hz); 7.22-7.33 (m, 4H, Ph).

c) 5,7-Bis [(1H-tetrazol-5-yl) methoxy] -3- (4-chlorobenzyl) -4-methyl-2H-1-benzopyran-2-one

The product of the previous example (0.40 g), sodium azide (0.14 g) and ammonium chloride (0.11 g) were heated in DMF (2 mL) at 100 ° C for 2 hours. The product was isolated as in Example 1c. The yield was 0.40 g (82%).

00 0 0 · 10

440 4 4 00 0

0 99049 9

9 0 08 0 0 0 0 09 4

490 44 9 0400

0000 00 000 00 00 1 H-NMR (DMSO-d ₆ , 300 MHz): 2.46 (s, 3H, CH ₃ ), 3.92 (s, 2H, CH ₂ ), 5.602 (s, 2H, OCH) ₂ Tet), 5.609 (s, 2H, OCH ₂ Tet), 6.83 (d, 1H, J = 2.5 Hz), 6.85 (d, 1H, J = 2.5 Hz), 7.20 -7.33 (m, 4H, Ph).

Example 15

Preparation of 5,7-bis [(1H-tetrazol-5-yl) methoxy] -3- (4-nitrobenzyl) -4-methyl-2H-1-benzopyran-2-one

(a) 5,7-Dihydroxy-4-methyl-3- (4-nitrobenzyl) -2H-1-benzopyran-2-one

A solution of florglucin (0.48 g) and ethyl 2- (4-nitrobenzyl) acetoacetate (1.00 g) in ethanol (150 mL) was treated with dry HCl at 0 ° C for 7.5 hours, and the solution was stored at this temperature overnight. The solvent was evaporated and the precipitate was washed with water. The yield was 0.63 g (51%). Melting point 280285 ° C.

1 H-NMR (DMSO-d ₆ , 300 MHz): 2.53 (s, 3H, CH ₃ ), 4.03 (s, 2H, CH ₂ ), 6.19 (d, 1H, j = 2, 4 Hz), 6.29 (d, 1H, J = 2.4 Hz), 7.40-7.51 and 8.11 and 8.17 (m, 4H, Ph), 10.25 (s, 1H, OH), 10.52 (s, 1H, OH).

b) 5,7-Bis (cyanomethoxy) -3- (4-nitrobenzyl) -4-methyl-2 H -1-benzopyran-2-one

The product of the previous example (0.57 g), chloroacetonitrile (0.27 g) and potassium carbonate (1.20 g) were heated in DMF (2 mL) at 100 ° C for 50 minutes.

• 9,999 9

9 9 999

9 9 9

9 9 9 «

9999 99 999

9 9 9

9 9 9

9 9 9

9 9 9

99

The product was isolated as described in Example 1b. The yield was 0.47 g (67%). Melting point 178-185 ° C.

1 H-NMR (DMSO-d ₆ , 400 MHz): 2.53 (s, 3H, CH ₃ ), 4.11 (s, 2H, CH ₂ ), 5.319 (s, 2H, OCH ₂ CN), 5.323 (s, 2H, OCH ₂ CN), 6.83 (d, 1H, J = 2.4 Hz),

6.96 (d, 1H, J = 2.4 Hz), 7.48-7.53 and 8.12-8.16 (m, 4H, Ph).

c) 5,7-Bis [(1H-tetrazol-5-yl) methoxy] -3- (4-nitrobenzyl) -4-methyl-2H-1-

The product of the previous example (0.38 g), sodium azide (0.12 g) and ammonium chloride (0.11 g) were heated in DMF (3 mL) at 100 ° C for 2 hours. The product was isolated as described in Example 1c. The yield was 0.25 g (54%). Mp 240-244 ° C.

1 H-NMR (DMSO-d ₆ , 400 MHz): 2.47 (s, 3H, CH ₃ ), 4.08 (s, 2H, CH ₂ ), 5.611 (s, 2H, OCH ₂ Tet), 5.673 (s, 2H, OCH ₂ Tet), 6.85 (d, 1H, J = 2.4 Hz),

6.87 (d, 1H, J = 2.4 Hz), 7.46-7.50 and 8.12 and 8.16 (m, 4H, Ph).

Example 16

Preparation of 5,7-bis [(1H-tetrazol-5-yl) methoxy] -3-cyclopentyl-4-methyl-2H-1-benzopyran-2-one

(a) 3-Cyclopentyl-5,7-dihydroxy-4-methyl-2H-1-benzopyran-2-one

HIM

O

Φ Φ φ φ φ φ φ φ φ φ φ φ φ φ φ φ φ φ φ φ φ φ φ

Λ) φ φ φ φ φ φ * Χ)) Χ

A solution of florglucinol (2.00 g) and ethyl 2-cyclopentylacetoacetate (3.14 g) in ethanol (40 mL) was treated with dry HCl at 0 ° C for 2.5 hours and the solution was kept at this temperature overnight. The solvent was evaporated and the precipitate was purified by flash chromatography washing with toluene / EtOAc-AcOH (8: 1: 1). The yield was 1.22 g (29%).

1 H-NMR (DMSO-d ₆ , 300 MHz): 1.50-1.88 (m, 8H, - (CH ₂ ) ₄ -), 2.57 (s,

3H, _CH3), 3.25 (m, 1H, CH), 6.11 (d, 1H, J = 2.4 Hz), 6.25 (d, 1H, J = 2.4 Hz), 10 25 (b, 2H, OH).

b) 5,7-Bis (cyanomethoxy) -3-cyclopentyl-4-methyl-2H-1-benzopyran-2-one

The product of the previous example (0.50 g), chloroacetonitrile (0.31 g) and potassium carbonate (0.61 g) were heated in DMF (2 mL) at 80 ° C for 40 minutes. The product was isolated as described in Example 1b. The yield was 0.56 g (86%).

1 H-NMR (DMSO-d ₆ , 300 MHz): 1.55-1.90 (m, 8H, - (CH ₂ ) ₄ -), 2.56 (s, 3H, CH ₃ ), 3.37 (m, 1H, CH), 5.29 (s, 2H, OCH ₂ CN), 5.31 (s, 2H, OCH ₂ CN), 6.75 (d, 1H, J = 2.5 Hz), 6.88 (d, 1H, J = 2.5Hz).

c) 5,7-Bis [(1H-tetrazol-5-yl) methoxy] -3-cyclopentyl-4-methyl-2H-1-benzopyran-2-one

The product of the previous example (0.30 g), sodium azide (0.13 g) and ammonium chloride (0.11 g) were heated in DMF (1 mL) at 100 ° C for 1.5 hours.

• ♦ 9 9 9 9

9

9999 99 999

The product was isolated as described in Example 1c. The yield was 0.30 g (80%). Mp 248-252 ° C.

1 H-NMR (DMSO-d ₆ , 400 MHz): 1.53-1.89 (m, 8H, - (CH ₂ ) ₄ -), 2.51 (s, 3H, CH ₃ ), 3.34 (m, 1H, CH), 5.59 (s, 2H, OCH ₂ Tet), 5.61 (s, 2H, OCH ₂ Tet), 6.80 (s, 2H).

Example 17

Preparation of 5,7-bis [(1H-tetrazol-5-yl) methoxy] -4-methyl-3- (1-naphthylmethyl) -2H-1-benzopyran-2-one

a) 5,7-Dihydroxy-4-methyl-3- (1-naphthylmethyl) -2H-1-benzopyran-2-one

A solution of florglucinol (0.47 g) and ethyl 2- (1-naphthylmethyl) acetoacetate (1.00 g) in ethanol (20 mL) was treated with dry HCl at 0 ° C for 3 hours and the solution was kept at this temperature overnight. The solvent was evaporated and the precipitate was washed with water and recrystallized from isopropanol-water (1: 1). The yield was 0.96 g (78%). M.p. 275-280 ° C.

1 H-NMR (DMSO-d ₆ , 400 MHz): 2.45 (s, 3H, CH ₃ ), 4.32 (s, 2H, CH ₂ ), 6.23 (d, 1H, J = 2, 5 Hz), 6.32 (d, 1H, J = 2.5 Hz), 6.97-8.25 (m, 7H, Naph), 10.26 (s, 1H, OH), 10.53 ( s, 1H, OH).

b) 5,7-Bis (cyanomethoxy) -4-methyl-3- (1-naphthylmethyl) -2H-1-benzopyran-2-one

·

444 ·

4 • 444 • 4 44 • 4 4 4 4

4 4 4 4 4 · · 4 4 4 4 4 »··· 44 * 44

The product of the previous example (0.80 g), chloroacetonitrile (0.36 g) and potassium carbonate (0.66 g) were heated in DMF (4 mL) at 100 ° C for 1 hour. The product was isolated as in Example 1b. The yield was 0.30 g (30%).

1 H-NMR (DMSO-d ₆ , 300 MHz): 2.45 (s, 3H, CH ₃ ), 4.40 (s, 2H, CH ₂ ), 5.34 (s, 2H, OCH ₂ CN) , 4.36 (s, 2H, _OCH2 CN), 6.86 (d, 1H, J = 2.5 Hz), 7.010 (d, 1H, J = 2.5 Hz), 7.016 to 8.27 ( m, 7H, Naph).

c) 5,7-bis [(1H-tetrazol-5-yl) methoxy] -4-methyl-3- (1-naphthylmethyl) -2H-1-benzopyran-2-one

The product of the previous example (0.25 g), sodium azide (0.080 g) and ammonium chloride (0.072 g) were heated in DMF (2 mL) at 100 ° C for 2.5 hours. The product was isolated as described in Example 1c. The yield was 0.11 g (36%). Melting point 164-174 ° C.

1 H-NMR (DMSO-d ₆ , 300 MHz): 2.40 (s, 3H, CH ₃ ), 4.37 (s, 2H, CH ₂ ), 5.63 (s, 2H, OCH ₂ Tet) 5.65 (s, 2H, OCH ₂ Tet), 6.87 (d, 1H, J = 2.5 Hz), 6.92 (d, 1H, J = 2.5 Hz), 6.98- 8.26 (m, 7H, Naph).

Example 18

Preparation of 1-benzyl-5,7-bis [(1H-tetrazol-5-yl) methoxy] -4-methyl-2H-1-quinolinone

a) 5,7-Dimethoxy-4-methyl-2 (1H) -quinolinone

Tert-Butylacetoacetate (1.58 g) was heated to 120 ° C and 3,5-dimethoxyaniline (1.53 g) dissolved in xylene (4 mL) was added. The mixture was heated. The mixture was heated to 9 ° C. 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 120 ° C for 20 minutes and then cooled to room temperature.

Methanesulfonic acid (2 mL) was added and the mixture was stirred at ambient temperature for 10 minutes. Water (40 mL) was added and the precipitate was filtered and dried. The yield was 1.31 g (60%).

1 H-NMR (DMSO-d ₆ , 300 MHz): 2.50 (s, 3H, CH ₃ ), 3.79 (s, 3H, OCH ₃ ),

3.83 (s, 3H, _OCH3), 6.03 (s, 1H, CH = C), 6.31 (d, 1H, J = 2.3 Hz), 6.45 (d,

1 H, J = 2.3 Hz), 11.4 (b, 1H, NH).

b) 1-Benzyl-5,7-dimethoxy-4-methyl-2 (1H) -quinolinone

The product of the previous example (1.20 g) was suspended in DMSO (15 mL) and t-BuOK (0.68 g) and benzyl bromide (1.03 g) were added. The reaction mixture was stirred at ambient temperature overnight. Water was added and the product was extracted into EtOAc. The EtOAc was dried and evaporated to dryness. The product was recrystallized from toluene. The yield was 0.80 g (47%).

1 H-NMR (DMSO-d ₆ , 300 MHz): 2.55 (d, 3H, J = 1.1 Hz, CH ₃ ), 3.71 (s, 3H, OCH ₃ ), 3.84 (s 3H, OCH ₃ ), 5.48 (b, 2H, NCH ₂ ), 6.29 (d, 1H, J = 1.1Hz, CH = C), 6.4 (s, 2H), 7, 18-7.33 (m, 5H, Ph).

c) 1-Benzyl-5,7-dihydroxy-4-methyl-2 (H) -quinolinone

• ·

The product of the previous example (0.69 g) was dissolved in CH ₂ Cl ₂ (14 mL) and the reaction mixture was cooled to -20 ° C. BBr ₃ (2.4 g) in CH ₂ Cl ₂ (1M solution) was added and the mixture was allowed to cool to ambient temperature overnight. The precipitate was filtered off, washed with CH ₂ Cl ₂ and dissolved in EtOAc. The EtOAc was washed with dilute HCl, dried and evaporated to dryness. The yield was 0.34 g (54%).

1 H-NMR (DMSO-d ₆ , 300 MHz): 2.56 (d, 3H, J = 1.0 Hz, CH ₃ ), 5.33 (b, 2H, NCH ₂ ), 6.11 (d 1 H, J = 2.1 Hz), 6.13 (d, 1H, J = 1.0 Hz, CH = C), 6.17 (d, 1H, J = 2.1 Hz), 7.12 7.34 (m, 5H, Ph), 9.90 (b, 1H, OH), 10.22 (s, 1H, OH).

d) 1-Benzyl-5,7-bis (cyanomethoxy) -4-methyl-2 (1H) -quinolinone

The product of the previous example (0.34 g), chloroacetonitrile (0.13 g) and potassium carbonate (0.34 g) were heated in DMF (2 mL) at 100 ° C for 1.5 hours. Water was added and the precipitate was filtered off and dried. The product was recrystallized from isopropanol. The yield was 0.20 g (46%).

1 H-NMR (DMSO-d ₆ , 400 MHz): 2.57 (s, 3H, CH ₃ ), 5.22 (s, 2H, OCH ₂ CN), 5.30 (s, 2H, OCH ₂ CN) ), 5.50 (b, 2H, NCH ₂ ), 6.42 (s, 1H, CH = C), 6.70 (d, 1H, J = 2.1 Hz), 6.73 (d, 1H J = 2.1 Hz), 7.21-7.32 (m, 5H, Ph).

e) 1-Benzyl-5,7-bis [(1H-tetrazol-5-yl) methoxy] -4-methyl-2H-1-quinolinone

O'

3 ······. «·. · ·

The product of the previous example (0.20 g), sodium azide (0.072 g) and ammonium chloride (0.060 g) were heated in DMF (2 mL) at 100 ° C for 3 hours. The product was isolated as described in Example 1c. The yield was 0.21 g (85%). Mp 246-249 ° C.

^! 1 H-NMR (DMSO-d ₆ , 400 MHz): 2.50 (s, 3H, CH ₃ ), 5.48 (b, 4H, OCH ₂ Tet, NCH ₂ ), 5.60 (s, 2H, OCH) ₂ Tet), 6.34 (s, 1H, CH = C), 6.64 (d, 1H, J = 1.9 Hz), 6.77 (d, 1H, J = 1.9 Hz), 7 18-7.32 (m, 5H, Ph).

Example 19

Preparation of 1-benzyl-5,7-bis [(1H-tetrazol-5-yl) methoxy] -3- (2-fluorobenzyl) -4-methyl-2 (1H) -quinolinone

a) 5,7-Dimethoxy-3- (2-fluorobenzyl) -4-methyl-2 (1H) -quinolinone

Ethyl 2- (2-fluorobenzyl) acetoacetate (2.5 g) in xylene (1 mL) was heated to 150 ° C and 3,5-dimethoxyaniline (1.46 g) in xylene was added in small portions over 30 minutes. . The reaction mixture was heated to 160 ° C for 3 hours and then cooled to room temperature. Methanesulfonic acid (1.7 mL) was added and the mixture was stirred at ambient temperature for 30 minutes. Water was added and the precipitate was filtered off and dried. The product was washed with hot ethanol. The yield was 0.64 g (21%).

1 H-NMR (DMSO-d ₆ , 300 MHz): 2.45 (s, 3H), 3.79 (s, 3H), 3.82 (s, 3H), 3.97 (s, 2H), 6.33 (d, 1H, J = 2.4 Hz), 6.48 (d, 1H, J = 2.4 Hz), 6.90-7.25 (m, 4H), 11.61 (s , IH).

b) 1-Benzyl-5,7-dimethoxy-3- (2-fluorobenzyl) -4-methyl-2 (1H) quinolinone

The product of the previous example (0.62 g) was treated with t-BuOK (0.23 g) and benzyl bromide (0.36 g) in DMSO (12 mL) at 60 ° C for 2.5 hours. The product was isolated as described in Example 18b. The yield was 0.39 g (49%).

1 H-NMR (DMSO-d ₆ , 300 MHz): 2.51 (s, 3H), 3.72 (s, 3H), 3.84 (s, 3H). 4.11 (s, 2H), 5.55 (b, 2H), 6.433 (d, 1H, J = 2.1 Hz), 6.443 (d, 1H, J = 2.1 Hz), 6.97- 7.33 (m, 9H).

c) 1-Benzyl-5,7-dihydroxy-3- (2-fluorobenzyl) -4-methyl-2 (1H) -quinolinone

The product of the previous example (0.34 g) was treated with BBr ₃ (8.48 g) in CH ₂ Cl ₂ (7 mL) as described in Example 18c. The yield was 0.30 g (82%).

1 H-NMR (DMSO-d ₆ , 400 MHz): 2.55 (s, 3H), 4.06 (s, 2H), 5.40 (b, 2H), 6.13 (d, 1H, J) = 2.1 Hz), 6.97-7.33 (m, 9H), 10.3 (b, 2H).

d) 1-Benzyl-5,7-bis (cyanomethoxy) -3- (2-fluorobenzyl) -4-methyl-2 (1H) -quinolinone

• to to - to • to

The product of the previous example (0.21 g), chloroacetonitrile (0.086 g) and potassium carbonate (0.37 g) were heated in DMF (2 mL) at 100 ° C for 2 hours. The product was isolated as described in Example 1b. The yield was 0.18 g (71%). 1 H-NMR (DMSO-d ₆ , 400 MHz): 2.53 (s, 3H), 4.13 (s, 2H), 2.53 (s, 2H) 5.29 (s, 2H), 5.57 (b, 2H), 6.746 (d, 1H, J = 2.3 Hz), 6.756 (d, 1H, J = 2.3 Hz), 7, 00-7.32 (m, 9 H).

e) 1-Benzyl-5,7-bis [(1H-tetrazol-5-yl) methoxy] -3- (2-fluorobenzyl) -4-methyl-2 (1H) -quinolinone

The product of the previous example (0.17 g), sodium azide (0.051 g) and ammonium chloride (0.042 g) were heated in DMF at 100 ° C for 3 hours, the product was isolated as described in Example 1c. The yield was 0.17 g (85%). Melting point 135-140 ° C.

1 H-NMR (DMSO-d ₆ , 400 MHz): 2.46 (s, 3H), 4.10 (s, 2H), 5.48 (s, 2H), 5.51 (b, 2H), 5.59 (s, 2H), 6.68 (d, 1H, J = 2.2 Hz), 6.79 (d, 1H, J = 2.2 Hz), 6.99-7.32 (m , 9H).

Example 20

Preparation of 1-benzyl-5,7-bis [(1H-tetrazol-5-yl) methoxy] -4-methyl-3- (2-phenylethyl) -2 (1H) -quinolinone

a) 5,7-Dimethoxy-4-methyl-3- (2-phenylethyl) -2 (1H) -quinolinone

O'

O

Ethyl 2- (2-phenylethyl) acetoacetate (2.70 g) in xylene (5 mL) was treated with 3,5-dimethoxyaniline (1.60 g) at 150 ° C as described in Example 19a. Methanesulfonic acid (4.0 mL) was added at room temperature and the mixture was heated to 80 ° C for 1 hour. The product was isolated as described in Example 19a. (Yield 1.38 g (41%)).

1 H-NMR (DMSO-d ₆ , 400 MHz): 2.45 (s, 3H), 2.64-2.68 (m, 2H); 2.822.86 (m, 2H), 3.78 (s, 3H), 3.81 (s, 3H), 6.30 (d, 1H, J = 2.3 Hz), 6.454 (d, 1H, J = 2.3 Hz), 7.18-7.30 (m, 5H), 11.45 (s, 1H).

b) 1-Benzyl-5,7-dimethoxy-4-methyl-3- (2-phenylethyl) -2 (1H) -quinolinone

The product of the previous example (0.61 g), tert-BuOK (0.24 g) and benzyl bromide (0.36 g) were heated in DMSO (12 mL) at 60 ° C for 2 hours. The product was isolated as described in Example 18b. The yield was 0.31 g (40%).

1 H-NMR (DMSO-d ₆ , 400 MHz): 2.51 (s, 3H), 2.73-2.77 (m, 2H), 2,963.00 (m, 2H), 3.70 (s 3H), 3.83 (s, 3H), 5.55 (b, 2H), 6.40 (s, 2H), 7.17-7.33 (m, 10H).

c) 1-Benzyl-5,7-dihydroxy-4-methyl-3- (2-phenylethyl) -2 (1H) -quinolinone

• · · ·

C0

F «· <

F · · I · · ·

The product of the previous example (0.31 g) was treated with BBr ₃ (0.75 g) in CH ₂ Cl ₂ (5 mL), as in Example 18c. The yield was 0.26 g (89%).

1 H-NMR (DMSO-d ₆ , 300 MHz): 2.56 (s, 3H), 2.69-2.75 (m, 2H), 2,902.95 (m, 2H), 5.39 (b (2H), 6.08 (d, 1H, J = 2.0 Hz), 6.19 (d, 1H, J = 2.0 Hz), 7.11-7.33 (m, 10H), 10 2 (b, 2H).

d) 1-Benzyl-5,7-bis (cyanomethoxy) -4-methyl-3- (2-phenylethyl) -2 (1H) -quinolinone

The product from the previous example (0.22 g). chloroacetonitrile (0.091 g) and potassium carbonate (0.39 g) were heated at 100 ° C for 2 hours. The product was isolated as in Example 1b. The yield was 0.20 g (76%).

1 H-NMR (DMSO-d ₆ , 400 MHz): 2.50 (s, 3H), 2.73-2.77 (m, 2H), 2,983.02 (m, 2H), 5.21 (s 2H, 5.29 (s, 2H), 5.56 (b, 2H), 6.70 (d, 1H, J = 2.1 Hz), 6.72 (d, 1H, J = 2, 1 Hz), 7.18-7.33 (m, 10H).

e) 1-Benzyl-5,7-bis [(1H-tetrazol-5-yl) methoxy] -4-methyl-3- (2-phenylethyl) -2 (1H) -quinolinone

The product of the previous example (0.19 g), sodium azide (0.057 g) and ammonium chloride (0.047 g) were heated in DMF at 100 ° C for 3 hours. The product was isolated as described in Example 1c. The yield was 0.18 g (78%). Mp 215-218 ° C.

1 H-NMR (DMSO-d ₆ , 400 MHz): 2.46 (s, 3H), 2.70-2.74 (m, 2H), 2.952.99 (m, 2H), 5.47 (s (2H), 5.54 (b, 2H), 5.57 (s, 2H), 6.64 (d, 1H, J = 2.0 Hz), 6.77 (d, 1H, J = 2, 0 Hz), 7.16-7.33 (m, 10H).

Example 21

Preparation of 5,7-bis (aminocarbonylmethoxy) -1,3-dibenzyl-4-methyl-2 (1H) -quinolinone

A mixture of 5,7-dihydroxy-1,3-dibenzyl-4-methyl-2 (1H) -quinolinone (0.5 g), potassium carbonate (0.9 g) and 2-chloroacetamide (0.25 g) in DMF (6.5 mL) was reacted at 100 ° C for two hours. The reaction mixture was treated with ice water and filtered. The product was washed with hot ethanol. Yield 0.32 g. Mp 252-253 ° C.

1 H-NMR (DMSO-d ₆ , 400 MHz): 2.63 (s, 3H, CH ₃ ), 4.13 (s, 2H, PhCH ₂ ), 4.37 (s, 2H, OCH ₂ ), 4.55 (s, 2H, OCH ₂ ), 5.54 (s, 2H, NCH ₂ Ph), 6.40 (d, 1H, J = 2Hz, ArH), 6.53 (d, 1H, J = 2 Hz, Ar- H), 7.13 to 7.33 (m, 10 H, Ph), 7.44 (d, 2H, J = 65 Hz, CONH _2), 7.47 (d, 2H, J = 68 Hz, CONH _2).

• · ♦ · φ · · φ · · ♦ '

Example 22

Preparation of 5,7-bis (ethoxycarbonylmethoxy) -1,3-dibenzyl-4-methyl-2 (1H) quinolinone

A mixture of 5,7-dihydroxy-1,3-dibenzyl-4-methyl-2 (1H) -quinolinone (1 g), ethyl 2-bromoacetate (0.63 ml) and potassium carbonate (1.49 g) in DMF (5) ml) was heated under nitrogen at 110 ° C for three hours, poured into ice water and filtered. The solid obtained was washed with ether and filtered again. The yield was 1.03 g, mp 113-116 ° C.

1 H-NMR (DMSO-d ₆ , 400 MHz): 1.15 (t, 3H, CH ₃ CH ₂ , J = 7.1 Hz), 1.20 (t, CH ₃ CH ₂ , J = 7, 1 Hz), 2.63 (s, 3H, CH ₃ ), 4.03 (q, 2H, CH ₂ CH ₃ , J = 7.1 Hz), 4.13 (s, 2H, CH ₂ Ph), 4.17 (q, 2H, CH ₂ CH _3, J = 7.1 Hz), 4.78 (s, 2H, _OCH2), 4.90 (s, 2H, _OCH2), 6.41 (d 1 H, J = 2.2 Hz), 6.44 (d, 1H, J = 2.2 Hz), 7.137.33 (m, 10H, Ph).

Example 23

Preparation of 5,7-bis (hydroxyaminocarbonylmethoxy) -1,3-dibenzyl-4-methyl-2 (1H) -quinolinone

OH

··

* «·

The product of the previous example (0.3 g), hydroxylamine hydrochloride (0.32 g) and 5 N NaOH (1.05 ml) were reacted in ethanol (8 ml) at 50 ° C for 6 hours. The reaction mixture was treated with water, adjusted to basic pH (pH 10) and filtered. The filtrate was acidified to pH 2 and filtered. The yield was 0.2 g, mp 121127 ° C.

H-NMR (DMSO-d _6, 400 MHz): tatutomerní forms of hydroxamic acid are apparent in _OCH2 -signal: 2.63 (s, 3H, _CH3), 4.13 (s, 2H, CH ₂ Ph), 4.41 (s, 2H, OCH ₂ ), 4.54 (s, 2H, OCH ₂ ), 4.64 (s, 2H, HON = C (OH) CH ₂ O), 4.65 (s, 2H , HON = C (OH) CH ₂ O), 4.77 (s, 2H, HON = C (OH) CH ₂ O), 4.78 (s, 2H, HON = C (OH) CH ₂ O), 5.54 (s, 2H, NCH ₂ Ph), 6.38-6.54 (m, 2H, ArH), 7.14-7.34 (m, 10H, Ph), 9.05 (b, 2H, NOH), 10.84 (s, 1H, HONHCO), 10.88 (s, 1H, HONHCO).

Example 24

Preparation of 5,7-bis [1- (6-hydroxypyridazinyl)] oxy-1,3-dibenzyl-4-methyl-2 (1H) -quinolinone

(a) 5,7-Bis [1- (6-chloropyridazinyl)] oxy-1,3-dibenzyl-4-methyl-2 (1H) -quinolinone

A mixture of 1,3-dibenzyl-5,7-dihydroxy-4-methyl-2 (1H) -quinolinone (0.5 g), 3,6-dichloropyridazine (0.83 g) and potassium carbonate (0.75 g) in DMF (12.5 mL) was stirred at 80 ° C for 4 hours. The reaction mixture was treated with water at pH 8 and filtered. The solids were recrystallized from ethanol-DMF (2: 1). The yield was 0.5 g. Mp 208-218 ° C.

»* ·» ^! 1 H-NMR (DMSO-d ₆ , 300 MHz): 2.43 (s, 3H, CH ₃ ), 4.16 (s, 2H,

CH ₂ Ph), 5.58 (s, 2H, NCH ₂ Ph), 7.09-7.33 (m, 12H, ArH + Ph), 7.55 (d, 1H, PyridH, J = 9.2 Hz), 7.70 (d, 1H, PyridH, J = 9.2 Hz), 7.93 (d, 1H, PyridH, J = 9.2 Hz), 7.98 (d, 1H, PyridH, J = 9.2 Hz).

b) 5,7-bis [1- (6-hydroxypyridazinyl)] oxy-1,3-dibenzyl-4-methyl-2 (1H) -quinolinone

The product of the previous example (0.2 g) and sodium acetate (0.13 g) in acetic acid (5 ml) were refluxed for 4 hours. The mixture was evaporated, treated with water at pH 10 and filtered. The filtrate was acidified to pH 6 and filtered. The yield was 70 mg.

1 H-NMR (DMSO-d ₆ , 300 MHz): 2.47 (s, 3H, CH ₃ ), 4.15 (s, 2H, CH ₂ Ph), 5.55 (s, 2H, NCH ₂ ) 6.93-7.34 (m, 15H, PyrH + ArH + Ph), 7.47 (d, 1H, J = 10Hz), 12.25 (s, 1H, NH), 12.38 ( s, 1H, NH).

Claims (18)

in which R ¹ is hydrogen, alkyl, alkenyl, aryl, arylalkyl, hydroxyalkyl, haloalkyl, alkoxy, COR _10, CONR ₁₀ R, OR _10, S (O) _m R _10, NR ₁₀ COR _n, or NR ₁₀ R where R ₁₀ is hydrogen, alkyl, alkenyl, aryl, arylalkyl, hydroxyalkyl, haloalkyl, alkoxy or hydroxy, and R _n is hydrogen, alkyl, aryl, aralkyl, alkoxy, aryloxy, hydroxy or acyl, or in case that X is NR _U, may also R! be carboxyalkyl, R ⁶ is hydrogen, alkyl, alkenyl, aryl, arylalkyl, R ² and R ⁷ are hydrogen, alkyl, aryl, arylalkyl, alkenyl, COR ₁₀ , CONR ₁₀ R 11, halogen, trifluoromethyl, nitro or cyano, wherein R _{10 and} R ₁₁ are as defined above, R ₃ is hydrogen, alkyl, aryl or arylalkyl, A is alkyl or substituted alkyl, m is 0-2 and n is 1-3, Y represents O, NR _U or S, wherein R _H is as defined above, X represents O, S or NR j i where R is as defined above, R _4, R _s, R ₈ and R ₉ are each independently one of the following groups: or when X is NR wherein _U may also R _4, R _5, R ₈ and R ₉ independently mean HOOC-, R ₁₂ OOC-, H ₂ NCO- or HOHNCO-, where R ₁₂ is alkyl, arylalkyl or aryl and wherein each aryl radical as defined above, alone or as part of another group, may be substituted, and pharmaceutically acceptable salts and esters thereof. A compound according to claim 1, wherein said compound has formula (I) and R ² is hydrogen. A compound according to claim 2, wherein R ¹ is hydrogen, C 1-6. ₆ alkyl, c _{2. 6} alkenyl, C ₆ _ ₁₀ aryl, C _{7th 12} arylalkyl, Cj.ghydroxyalkyl, C _"6 haloalkyl or alkoxy. The compound of claim 3, wherein Y is O or S, and X is O. The compound of claim 3, wherein Y is O or S; and X is NR 11 where R 11 is hydrogen, C 1-6 alkyl, C ₆ . ₁₀ aryl, C ₇ . ₁₂ arylalkyl, Cj.galkoxyskupina, C _{6. 10} aryloxy, hydroxy, C _1-6 alkanoyl or C ₁ . ₆ carboxyalkyl. 6. A compound according to claim 5 wherein R3 is hydrogen, C ^ alkyl, _C6 .I "aryl or C _{7th 12} arylalkyl. The compound of claim 6, wherein R 3 is C 1-4 alkyl. A compound according to claim 7, wherein A is straight chain or branched. 4 4 C 1-4 alkylene and R ⁴ and R ⁵ are each O / N-N H; or wherein X is X is NR _11, then R ₄ and R ₅ may also be HOOC-, R ₁₂ OOC-, H ₂ NCO- or HOHNCO-, wherein R ₁₂ is C 11. ₆ alkyl, C _{6. 10} aryl, or C _{7th 12} arylalkyl. The compound of claim 1, wherein said compound has the formula (II) and R 7 is hydrogen. A compound according to claim 9, wherein R 6 is hydrogen, C 1-4 alkyl, C ₆ . ₁₀ aryl, C ₇ . ₁₂ arylalkyl; and n is 1, 2 or 3 The compound of claim 10, wherein Y is O or S, and X is O. The compound of claim 10, wherein Y is O or S; and X is NR11, wherein R11 is hydrogen, C1-6alkyl, _C6 . ₁₀ aryl, C ₇ . ₁₂ arylalkyl, Cj.galkoxyskupina, C _{6. 10} aryloxy, hydroxy, or C₁₋₄ Cj.galkanoyl carboxyalkyl. The compound of claim 11, wherein A is a straight chain or branched C 1. ₄ alkylene. A compound according to claim 13, wherein R 4 and R 5 are each ^H 3 ^C X, N N 1 // N-N / • fr 9 9 99 frfrfr • <fr · fr · fr; or wherein X is X is NR _1b then it may also be R ₄ and R 5 _with HOOC-, R ₁₂ OOC-, H ₂ NCO- or HOHNCO-, wherein R ₁₂ is C ₁ . ₆ alkyl, C _{6. 10} aryl, or C _{7th 12} arylalkyl. 15. A pharmaceutical composition comprising a compound of claim 1 as an active ingredient, together with a pharmaceutically acceptable carrier. A method of treating heart failure comprising administering to a subject in need thereof a therapeutically effective amount of a compound of claim 1. A method of treating and preventing myocardial paralysis comprising administering to a subject in need thereof a therapeutically effective amount of a compound of claim 1. 18. Compound of formula (XXV): where R ¹ is hydrogen, alkyl, alkenyl, aryl, arylalkyl, hydroxyalkyl, haloalkyl, alkoxy, CORI _0, CONR ₀ R, OR _10, S (O) _m R _10, NR ₁₀ COR _n, or NR ₁₀ R _n, wherein R ₁₀ is hydrogen, alkyl, alkenyl, aryl, arylalkyl, hydroxyalkyl, haloalkyl, alkoxy or hydroxy, and R _n is hydrogen, alkyl, aryl, aralkyl, alkoxy, aryloxy, hydroxy or acyl, or carboxyalkyl, • 5 R is hydrogen, alkyl, aryl or arylalkyl, R ¹¹ is hydrogen, alkyl, aryl, arylalkyl, alkoxy, aryloxy, hydroxy or acyl, or carboxyalkyl, and wherein each aryl residue as defined above, alone or as part of another group, may be substituted.