HK1095093B - Nitrooxy derivatives of losartan, valsatan, candesartan, telmisartan, eprosartan and olmesartan as angiotensin-ii receptor blockers for the treatment of cardiovascular diseases - Google Patents

Description

Nitrooxy derivatives of losartan, valsartan, candesartan, telmisartan, eprosartan and olmesartan as angiotensin II receptor blockers for the treatment of cardiovascular diseases

The present invention relates to angiotensin II receptor blocker (ARB) derivatives. More particularly, the present invention relates to ARB nitro derivatives, pharmaceutical compositions containing them and their use in the treatment of cardiovascular, renal and chronic liver disease, inflammation (inflammatory processes) and metabolic syndrome.

Angiotensin II receptor blockers refer to a class of compounds that mainly includes losartan, EXP3174, candesartan, telmisartan, valsartan, eprosartan, irbesartan, and olmesartan.

ARBs have only been approved for the treatment of hypertension, the antihypertensive activity of which is due primarily to the selective blockade of AT₁The receptor, and thus the angiotensin II, produces a hypotensive effect. Angiotensin II stimulates the synthesis and secretion of aldosterone and raises blood pressure through potent direct vasoconstriction.

Angiotensin II receptor blockers have been reported to have side effects such as hypotension, hyperkalemia, myalgia, respiratory diseases, nephropathy, backache, gastrointestinal disorders, fatigue and neutropenia (Martindale, 33 rd edition, page 921).

It is an object of the present invention to provide novel ARBs derivatives which are not only capable of eliminating or at least reducing the side effects associated with their parent compounds, but also possess high pharmacological activity. Unexpectedly, it was found that the angiotensin II receptor blocker nitro derivatives have a broader range of pharmacological activity and increased tolerability and a significantly improved overall profile compared to the parent (native) compound.

In particular, it has been found that the nitro derivatives of the angiotensin II receptor blockers of the present invention exhibit strong anti-inflammatory, antithrombotic and antiplatelet activities and are therefore useful for the treatment or prevention of heart failure, myocardial infarction, ischemic stroke, atherosclerosis, ocular and pulmonary hypertension, diabetic nephropathy, peripheral vascular disease, left ventricular dysfunction and hypertrophy, liver fibrosis, portal hypertension and metabolic syndrome.

Accordingly, an object of the present invention is the nitro derivatives of angiotensin II receptor blockers of general formula (I) and the pharmaceutically acceptable salts or stereoisomers thereof:

R-(Y-ONO₂)_s (I)

wherein:

s is an integer equal to 1 or 2;

r is selected from the group consisting of angiotensin II receptor blocker residues of the following formula (II) or (III):

wherein:

R₀is composed of

Or a group-N capable of binding to Y₀Which has one of the following meanings:

-COO-, -O-, -CONH-, -OCO-, -OCOO-or

Wherein R 'and R' are the same or different and are H or straight or branched C₁-C₄An alkyl group;

R₁selected from:

or

Wherein m is an integer equal to 0 or 1, N₀The definition is the same as above;

wherein N is₁Meaning of (A) and N₀Same or equal to-COOH;

provided that at least one N₁The group is equal to-COO-or-CONH-, i.e. it is a group capable of binding to Y;

y is a divalent group having the following meaning:

a)

-straight or branched C₁-C₂₀Alkylene, preferably C₁-C₁₀Optionally substituted with one or more substituents selected from: halogen atom, hydroxy group, -ONO₂Or T₀Wherein T is₀is-OC (O) (C)₁-C₁₀Alkyl) -ONO₂or-O (C)₁-C₁₀Alkyl) -ONO₂；

Cycloalkylene having 5 to 7 carbon atoms in the cycloalkylene ring, which ring is optionally substituted by a side chain T, where T is a linear or branched alkyl group having 1 to 10 carbon atoms, preferably CH₃；

Wherein n is an integer of 0 to 20, n¹Is an integer of 1 to 20;

d)

wherein:

n¹is as defined above, n²Is an integer of 0 to 2;

X₁-OCO-or-COO-, R²Is H or CH₃；

e)

Wherein:

n¹、n²、R²and X₁The definition is the same as above;

Y¹is-CH₂-CH₂-or-CH ═ CH- (CH)₂)_n ²-；

Wherein:

n¹and R²As defined above, R³Is H or-COCH₃；

Provided that-ONO is present when Y is selected from the divalent radicals mentioned under b) -f)₂Group and- (CH)₂)_n ¹Group attachment;

wherein X₂is-O-or-S-, n³Is an integer from 1 to 6, preferably from 1 to 4, R²The definition is the same as above;

wherein:

n⁴is an integer of 0 to 10;

n⁵is an integer of 1 to 10;

R⁴、R⁵、R⁶、R⁷identical or different and is H or straight-chain or branched C₁-C₄Alkyl, preferably R⁴、R⁵、R⁶、R⁷Is H;

wherein-ONO₂The radical being attached to

Wherein n is⁵The definition is the same as above;

Y²is a saturated, unsaturated or aromatic 5-or 6-membered heterocyclic ring containing one or more heteroatoms selected from nitrogen, oxygen, sulphur,

and is selected from

The term "C" as used herein₁-C₂₀Alkylene "means branched or straight chain C₁-C₂₀The hydrocarbon group preferably has 1 to 10 carbon atoms, such as methylene, ethylene, propylene, isopropylene, n-butylene, pentylene, n-hexylene, and the like.

The term "C" as used herein₁-C₁₀Alkyl "refers to a branched or straight chain alkyl group containing 1 to 10 carbon atoms and includes methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, pentylAlkyl, hexyl, octyl, and the like.

The term "cycloalkylene" as used herein refers to a ring having 5 to 7 carbon atoms, including but not limited to, optionally substituted with a side chain such as straight or branched (C)₁-C₁₀) -alkyl, preferably CH₃Substituted cyclopentylene, cyclohexylene.

The term "heterocycle" as used herein refers to a saturated, unsaturated or aromatic 5-or 6-membered ring containing one or more heteroatoms selected from nitrogen, oxygen, sulfur, such as pyridine, pyrazine, pyrimidine, pyrrolidine, morpholine, imidazole and the like.

Another aspect of the present invention provides the use of a compound of formula (I) in combination with at least one compound for the treatment of cardiovascular disease selected from the group consisting of: ACE inhibitors, HMGCoA reductase inhibitors, beta-adrenergic blockers, calcium channel blockers, diuretics, antithrombotic agents such as aspirin, nitrated (nitrosated) ACE inhibitors, nitrated HMGCoA reductase inhibitors, nitrated beta-adrenergic blockers, nitrated aspirin, and nitrated diuretics.

Suitable ACE inhibitors, HMGCoA reductase inhibitors, beta-adrenergic blockers, calcium channel blockers, antithrombotic agents and diuretics are described in The literature, for example, The Merck Index (13 th edition).

Suitable nitrated compounds are disclosed in WO 98/21193, WO 97/16405 and WO 98/09948.

It is reported that the above compounds can be administered simultaneously or sequentially.

The invention also provides a kit comprising one or more containers filled with one or more compounds and/or compositions of the invention and one or more of the above compounds for use in the treatment of cardiovascular disease.

As mentioned above, the invention also includes pharmaceutically acceptable salts of the compounds of formula (I) and stereoisomers thereof.

Examples of pharmaceutically acceptable salts are those formed with inorganic bases such as sodium, potassium, calcium and aluminum hydroxides, or with organic bases such as lysine, arginine, triethylamine, dibenzylamine, piperidine and other acceptable organic amines.

When the compounds of the present invention contain a salifiable nitrogen atom in the molecule, they may be converted to the corresponding salts by reaction with the corresponding organic or inorganic acid in an organic solvent such as acetonitrile, tetrahydrofuran.

Examples of organic acids are: oxalic acid, tartaric acid, maleic acid, succinic acid, citric acid. Examples of mineral acids are: nitric acid, hydrochloric acid, sulfuric acid, phosphoric acid. Salts with nitric acid are preferred.

Compounds of the invention having one or more asymmetric carbon atoms may exist as optically pure enantiomers, pure diastereomers, mixtures of enantiomers, mixtures of diastereomers, racemic mixtures of enantiomers, racemates or racemic mixtures. All possible isomers, stereoisomers and mixtures thereof of the compounds of formula (I) are within the scope of the present invention.

Preferred compounds are those of formula (I) wherein:

s and R are as defined above;

y is a divalent group having the following meaning:

a)

-straight or branched C₁-C₁₀Alkylene, optionally substituted by T₀And (3) substitution: wherein T is₀The definition is the same as above;

b)

wherein n is an integer equal to 0 or 1, n¹Is an integer equal to 1; provided that-ONO₂Group and- (CH)₂)_n ¹Radical is linked toConnecting;

wherein X₂is-O-or-S-, n³Is an integer equal to 1, R²Is H;

the following are preferred compounds of the invention:

as mentioned above, pharmaceutical compositions comprising at least one compound of formula (I) according to the invention together with non-toxic auxiliary substances and/or carriers conventionally used in the pharmaceutical field are also an object of the present invention.

The daily dose of active ingredient to be administered may be a single dose, or may be an effective amount divided into several smaller doses administered over the course of a day. Generally, the total daily dose may be an amount of preferably 50-500 mg. The dosage regimen and frequency of administration for the treatment of the above-mentioned diseases with the compounds of the present invention and/or the pharmaceutical compositions of the present invention will be selected in accordance with a variety of factors including, for example, the age, weight, sex and medical condition of the patient and the severity of the disease, the route of administration, pharmacological factors and, optionally, (eventual) combination therapy with other drugs. In some cases, dosage levels below or above the aforementioned ranges and/or more frequent may be appropriate and this is within the logical judgment of the physician and depends on the disease state.

The compounds of the invention may optionally be administered orally, parenterally, rectally or topically, by inhalation or aerosol formulation with conventional non-toxic pharmaceutically acceptable carriers, adjuvants and vehicles as required. Topical administration also involves the use of transdermal administration such as transdermal patches or iontophoresis devices. The term "parenteral" as used herein includes subcutaneous injections, intravenous, intramuscular, intrasternal injection or infusion techniques.

Injectable preparations may be formulated according to the known art using suitable dispersing or wetting agents and suspending agents, for example, sterile injectable aqueous or oleaginous suspensions. The sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally-acceptable diluent or solvent. Among the acceptable solvents and solvents are water, ringer's solution and isotonic sodium chloride. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose, any bland fixed oil may be employed including synthetic mono-and diglycerides, and in addition fatty acids such as oleic acid may be used in the preparation of injectables.

Suppositories for rectal administration of the active ingredient can be prepared by mixing the active ingredient with suitable non-irritating excipients such as cocoa butter and polyethylene glycols.

Solid dosage forms for oral administration may include capsules, tablets, pills, powders, granules and gels. In such solid dosage forms, the active compound may be mixed with at least one inert diluent, for example sucrose, lactose or starch. Such dosage forms may also contain, in accordance with common practice, other substances in addition to inert diluents, for example lubricating agents such as magnesium stearate. In the case of capsules, tablets and pills, the dosage forms may also comprise buffering agents. In addition, tablets and pills can be prepared with an enteric coating.

Liquid dosage forms for oral administration may include pharmaceutically acceptable emulsions, solutions, suspensions, syrups and elixirs containing inert diluents commonly used in the art, such as water. Such compositions may also contain adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and the like.

The compounds of the invention can be synthesized as follows:

A) when R is a residue of formula (II), a compound of general formula (I) as defined above or a pharmaceutically acceptable salt thereof:

R-(Y-ONO₂)_s (I)

obtainable by a process comprising:

i) reacting a compound of formula (IV):

R₂-(Y-Hal)_s (IV)

wherein s is 1, R₂Is a residue of formula (IIA):

wherein R is₃Is a radical of the formula (VA)

Wherein A ═ H or W, W is a tetrazole protecting group such as trityl, tert-Butoxycarbonyl (BOC) and ethoxycarbonyl, or R₃Is a-COO-group capable of binding to Y;

R₁selected from the groups (IIa) to (IIe) as defined above, wherein N₀Is a group capable of binding to Y;

y is as defined above and Hal is a halogen atom, preferably Cl, Br or I;

with AgNO in a suitable organic solvent such as acetonitrile or Tetrahydrofuran (THF) under nitrogen in the dark at a temperature in the range of 20 deg.C to 80 deg.C₃Carrying out reaction; alternatively, AgNO can be reacted with AgNO in a solvent such as acetonitrile or THF under microwave irradiation at a temperature in the range of 100-₃Reacting in a short time (1-60 min); and

ii) optionally acid hydrolysis of the tetrazole protecting group W as known in the art, e.g., as described in T.W. Greene, "Protective groups in organic synthesis", Harvard university Press, 1980; and

iii) if desired, converting the resulting compound of formula (I) into a pharmaceutically acceptable salt thereof.

A compound of formula (IV) may be prepared by reacting a compound of formula (V):

wherein R is₅Is a group of the formula (VA) or-COOH, R₄With R₁Are the same, and N₀(ii) a group of compounds selected from-COOH or-OH,

i.1) when R is₅When it is a (VA) group, R₄＝R₁And R is₁Is a group (IIa) in which m is 1, N₀＝-OH，

With a compound of formula (VI) or (VII):

Hal-Y-COAct (VI)

Hal-Y-OCOAct (VII)

where Hal and Y are as defined above, Act is Hal or a carboxylic acid activating group for peptide chemistry, such as:

the reaction is usually carried out in the presence of an inorganic or organic base in an aprotic polar/nonpolar solvent such as DMF, THF or CH₂Cl₂In the temperature range of 0-65 ℃; or in H₂O/Et₂In an O two-phase system, the reaction is carried out at the temperature of between 20 and 40 ℃;

compounds of formula (VI) in which Act ═ Hal are commercially available, or can be prepared from the corresponding acids of formula (VIII) by well-known reactions, for example by reaction with thionyl chloride or oxalyl chloride, P in inert solvents such as toluene, chloroform, DMF and the like^IIIOr P^vTo give:

Hal-Y-COOH (VIII)

the corresponding acid is a commercially available compound.

Compounds of formula (VI) wherein Act is other than Hal can be obtained from the corresponding compounds of formula (VI) wherein Act is Hal by reaction with N-hydroxysuccinimide or an appropriately substituted phenol in the presence of a base known from the literature.

Compounds of the formula (VII) in which Act Hal are commercially available or can be obtained from the corresponding alcohol of the formula (IX) by reaction with triphosgene in the presence of an organic base:

Hal-Y-OH (IX)

compounds of formula (VII) wherein Act is other than Hal can be obtained from the corresponding compounds (VII) wherein Act is Hal by reaction with N-hydroxysuccinimide or an appropriately substituted phenol in the presence of a base known from the literature.

Alternatively, compounds of formula (IV) can be obtained by reacting a compound of formula (V) as defined in i.1) with a commercially available compound of formula (VIII) as defined above in the presence of a condensing agent such as Dicyclohexylcarbodiimide (DCC), EDAC, in the presence of a catalytic amount of DMAP or an activating agent such as N, N' -Carbodiimidazole (CDI), in a solvent such as DMF, THF, chloroform at a temperature in the range of-5 ℃ to 50 ℃;

i.2) when R is₅Is a (VA) group or-COOH, R₄＝R₁And R is₁Is selected from the group consisting of where m is 0, N₀When the groups (IIa) to (IId) of — COOH are present,

in a solvent such as DMF, THF, chloroform at a temperature in the range of-5 ℃ to 50 ℃ with a compound of formula (IX) as defined above in the presence of a condensing agent such as Dicyclohexylcarbodiimide (DCC), EDAC, in the presence of catalytic amounts of DMAP or activation of the carboxyl group with a reagent such as N, N' -Carbodiimidazole (CDI).

The compounds of formula (IX) are commercially available. Alternatively, the-COOH group is converted to an activated acid chloride or to another group suitable for esterification according to methods well known in the literature and in the presence of an organic or inorganic base in an aprotic polar/non-polar solvent such as DMF, THF or CH₂Cl₂In the temperature range of 0-65 ℃; or in H₂O/Et₂In an O two-phase system, the esterification reaction is carried out at the temperature of 20-40 ℃;

al) or, when R is a residue of formula (II), a compound of formula (I) as defined above can be obtained by reacting a compound of formula (V) as defined above with:

1.1) when R is₅Is a (VA) group, R₄＝R₁And R is₁Is a group (IIa) in which m ═ 1 and N₀When the compound is represented by-OH,

with a compound of formula (X):

O₂NO-Y-COZ (X)

wherein Y is as defined above and Z is OH or a defined Act group,

the reaction is carried out using the most suitable synthetic route, for example in a solvent such as DMF, THF, chloroform, at a temperature in the range from-5 ℃ to 50 ℃ and/or in the presence of an organic or inorganic base, in the presence of a condensing agent such as Dicyclohexylcarbodiimide (DCC) or EDAC, or activated with N, N' -Carbonyldiimidazole (CDI).

The corresponding halogen derivatives of the formula (VI) or (VIII) can be reacted with AgNO from the corresponding alcohols by reaction with nitric acid and acetic anhydride at temperatures in the range from-50 ℃ to 0 ℃ or as described above₃Reacting to obtain the compound of formula (X).

i.2.1) when R₅Is a (VA) or-COOH group, R₄＝R₁And R is₁Is selected from the group consisting of wherein m is 0 and N₀When the groups (IIa) to (IId) of — COOH are present,

in a temperature range of-5 deg.C to 50 deg.C in a solvent such as DMF, T_HF. In chloroform, in the presence of a condensing agent such as Dicyclohexylcarbodiimide (DCC) or EDAC or an activating agent such as N, N' -Carbyldiimidazole (CDI),

with a compound of formula (XI):

O₂NO-Y-OH (XI)

wherein Y is as defined above.

A compound of formula (IX) may be prepared by reacting a compound of formula (IX) with AgNO in a suitable organic solvent such as acetonitrile or THF under nitrogen at a temperature in the range 20 ℃ to 80 ℃₃Reacting to obtain a compound of formula (XI); alternatively, AgNO may be reacted with AgNO in a solvent such as acetonitrile or THF under microwave irradiation at a temperature in the range of 100-₃The reaction is carried out in a short time (1-60 min).

Or, when R is₅Is a (VA) or-COOH group, R₄＝R₁And R is₁Is selected from the group consisting of wherein m is 0 and N₀When the group (IIa) to (IIe) of-COOH is present, the group (IIa) to (IIe) can react withWith a compound of formula (XI.1) in the presence of an inorganic or organic base salified by carboxyl groups:

O₂NO-Y-Hal (XI.1)

wherein Y and Hal are as defined above.

B) Can be prepared by reacting a compound of formula (XII):

R⁶-(Y-Hal)_s (XII)

wherein s is 2, R₆Is a residue (III), N₁is-COO-, Y and Hal are as defined above,

as described above with AgNO₃The reaction gives a compound of general formula (I) wherein R is a residue of formula (III).

Can be prepared by reacting a compound of formula (XIII):

as previously described in the temperature range of-5 ℃ to 50 ℃ in the presence of a condensing agent such as Dicyclohexylcarbodiimide (DCC) or EDAC or an activating agent such as N, N' -Carbodiimidazole (CDI) in a solvent such as DMF, THF, chloroform,

with a compound of formula (IX) as defined above to give a compound of formula (XII).

Or converting the-COOH group into an active acid chloride or into another group suitable for esterification, according to methods well known in the literature, and in the presence of an organic or inorganic base in an aprotic polar/nonpolar solvent such as THF or CH₂Cl₂In the temperature range of 0 ℃ to 65 ℃, or in a two-phase system.

B1) Alternatively, when R is a residue of formula (III), a compound of formula (I) as hereinbefore defined may be obtained by reacting a compound of formula (XIII) with a compound of formula (XI) as hereinbefore defined in the presence of a condensing agent or activating agent as hereinbefore defined.

Alternatively, the-COOH groups can be converted into salts using inorganic or organic bases according to methods well known in the literature and then reacted with:

O₂NO-Y-Hal (XI.1).

C) When s ═ 1, R is a residue of formula (II) wherein R is₀Is a tetrazolyl group, R₁Is a group (IIa) in which m ═ 1 and N₀Is composed of

Wherein R 'and R' are as defined above,

a compound of formula (I) as hereinbefore defined may be prepared by reacting a compound of formula (IVa):

R₂-(CR′R＂-Hal)_s (IVa)

wherein s is 1, R₂And Hal are as defined above, R₃Is a (VA) group, R₁Wherein m is 1 and N₀A (IIa) group which is-OCOO-,

with a compound of formula (X) as defined above in the presence of an organic or inorganic base in a polar solvent such as DMF, THF, acetonitrile at a temperature in the range of-5 ℃ to 60 ℃ or in a two-phase system known in the literature.

By reacting a compound of formula (V) as defined above, wherein R₅Is a (VA) group, R₄＝R₁，R₁Wherein m is 1 and N₀The (IIa) group of — OH,

with a compound of formula (VIIa) in the same manner as described for compound (IV):

Hal-CR′R＂-OCOAct (VIIa)

wherein Act has the same meaning as described in (VII) above;

the tetrazole protecting group is optionally hydrolyzed as described above to provide compound (IVa).

D) A compound of formula (I) as hereinbefore defined wherein when s ═ 1, R is a residue of formula (II) wherein R is₀Is a tetrazolyl group, R₁Selected from the group consisting of (IIa) - (IIc), wherein m ═ 0 and N₀Is composed of

Wherein R 'and R' are as defined above,

by reacting a compound of formula (I) wherein R is₅Is a (VA) group; r₄＝R₁And R is₁In which N is₀Compounds of formula (V) of the (IIc) group of ═ COOH in the presence of organic or inorganic bases, in polar solvents such as DMF, THF, acetonitrile, in the temperature range from-5 ℃ to 60 ℃ or in two-phase systems known in the literature,

with a compound of formula (XIV):

Hal-CR′R＂-OCOO-Y-ONO₂ (XIV)

wherein Hal, Y, R 'and R' are as defined above.

The compound of formula (XIV) can be obtained by reacting compound (XI) with compound (VIIa) as defined above. Usually in the presence of a base in an aprotic polar/nonpolar solvent such as THF or CH₂Cl₂In the temperature range of 0-65 ℃; or in H₂O/Et₂In an O two-phase system, the reaction is carried out at a temperature ranging from 20 ℃ to 40 ℃; optionally acid hydrolyzing the tetrazole protecting group as described above.

E) When s ═ 1, R is a residue of formula (II) wherein R is₀Is a tetrazolyl group, R₁Compounds of formula (I) as defined above, selected from the group consisting of (IIa) to (IIc), can also be obtained by reacting a compound of formula (XV) with a commercially available compound of formula (XVI):

wherein R is₇Are residues (IIa) - (IIc), R₃Hal is as defined above for the (VA) radical. Usually in the presence of a base in an aprotic polar/nonpolar solvent such as DMF, THF or CH₂Cl₂At a temperature in the range of-15 ℃ to 80 ℃; or in H₂O/Et₂In an O two-phase system, the reaction is carried out at a temperature ranging from 20 ℃ to 40 ℃; and optionally acid hydrolyzing the tetrazole protecting group as described above.

Can be prepared by reacting a compound of formula (XVII):

R₈-(Y-Hal)

(XVII)

wherein R is₈Is a residue of formula (IIa.1), (IIb.1) or (IIc.1):

wherein PG is an N-protecting group such as BOC or trityl,

as described above with AgNO₃Reaction, optionally hydrolysis of the N-protecting group, gives the compound of formula (XV).

Wherein R is₈Is (IIa.1); wherein m is 1 and N₀Compound (XVII) which is ═ OCO-can be obtained by reacting the corresponding alcohol with the aforementioned compound of formula (VI) or (VII).

The alcohols defined above are obtained from commercially available compounds of the formula (IIa.2) by known protection and reduction reactions:

wherein m is a number of 0 s,N₀₀is-CHO.

Wherein R is₈Is m ═ 0 and N₀(IIa.1) or R of ═ -COO-₈Is N₀Compounds (XVII) of formula (iib.1) or (iic.1) (-COO-) can be obtained by reacting the corresponding acid with the aforementioned compounds of formula (IX).

The corresponding acids of (IIa.1) as defined above may be prepared from those in which m is 0, N₀₀The compound (IIa.2) which is-CHO is obtained by known protection and oxidation reactions.

The corresponding acids as defined above for (IIb.1) and (IIc.1) are obtained from commercially available (IIb.2) and (IIc.2) by known protection reactions:

wherein N is₀is-COOH.

The following examples further illustrate the invention and do not limit it.

Example 1

4- (Nitropoxymethyl) benzoic acid 2-butyl-4-chloro-1- [ [2 '- (1H-tetrazol-5-yl) [1, 1' -biphenyl ] -4-yl ] methyl ] -1H-imidazol-5-methyl ester (corresponding to Compound (4))

Triphenylmethyl chloride (4.68g, 16.8mmol) was added portionwise to a solution of losartan potassium salt (7.0 g; 15.2mmol) in THF (150 ml). The resulting mixture was stirred at room temperature for 24 hours. The reaction was then adsorbed onto silica gel and purified by flash chromatography (n-hexane/AcOEt 6:4) to give 2-butyl-4-chloro-1- [ [2 '- (1-trityltetrazol-5-yl) [1, 1' -biphenyl ] -4-yl ] methyl ] -1H-imidazole-5-methanol (6.7g, 66%).

The title compound (4) can be obtained from this compound by two different synthetic methods:

synthesis Process A

To 2-butyl-4-chloro-1- [ [2 '- (1-trityltetrazol-5-yl) [1, 1' -biphenyl ] at 0 deg.C]-4-yl]Methyl radical]-1H-imidazole-5-methanol (1.7g, 2.6mmol), 4- (nitrooxymethyl) benzoic acid (0.66g, 3.38mmol) and N, N-dimethylaminopyridine (0.049g, 0.4mmol) in CH₂Cl₂(20ml) and THF (6ml) solution dicyclohexylcarbodiimide (0.722g, 3.50mmol) in CH was added slowly₂Cl₂(5ml) solution, the reaction was stirred at room temperature for 24 h. The dicyclohexylurea formed is then filtered off and the organic phase is concentrated. Purifying the crude material by silica gel chromatography (n-hexane/AcOEt 75:25) to obtain 4- (nitrooxymethyl) benzoic acid 2-butyl-4-chloro-1- [ [2 '- (1-trityltetrazol-5-yl) [1, 1' -biphenyl]-4-yl]Methyl radical]-1H-imidazole-5-methyl ester (1.2g, 55%) as a white solid.

2-butyl-4-chloro-1- [ [2 '- (1-trityltetrazol-5-yl) [1, 1' -biphenyl ] 4- (nitrooxymethyl) benzoic acid]-4-yl]Methyl radical]-1H-imidazole-5-methyl ester (1.2g, 1.42mmol) in CH₂Cl₂(10ml), HCl was bubbled through the solution for 20 min. The mixture was then concentrated and subjected to flash Chromatography (CH)₂Cl₂Acetone 8:2, followed by acetone) purification to give crude compound, which is dissolved in H₂O/CH₃CN, freeze drying to obtain 4- (nitrooxymethyl) benzoic acid 2-butyl-4-chloro-1- [ [2 '- (1H-tetrazol-5-yl) [1, 1' -biphenyl]-4-yl]Methyl radical]-1H-imidazole-5-methyl ester as a white solid (0.304g, 36%).

¹H-NMR(DMSO-d₆)：7.73-7.56(7H，m)；7.24(1H，d)；7.00(4H，m)；5.60(2H，s)；5.39(2H，s)；5.28(2H，s)；2.61(2H，t)；1.53(2H，m)；1.28(2H，m)；0.82(3H，t).

Synthesis method B

To 2-butyl-4-chloro-1- [ [2 '- (1-trityltetrazol-5-yl) [1, 1' -biphenyl ] at 0 deg.C]-4-yl]Methyl radical]-1H-imidazole-5-methanol (1.7g, 2.6mmol), 4- (chloromethyl) benzoic acid (0.571g, 3.35mmol) and N, N-dimethylaminoPyridine (0.049g, 0.4mmol) in CH₂Cl₂To a solution of (20ml) and THF (6ml) was added slowly dicyclohexylcarbodiimide (0.644g, 3.12mmol) and the reaction was stirred at room temperature for 24 h. The dicyclohexylurea formed is then filtered off and the organic phase is concentrated. The crude material was purified by flash chromatography (n-hexane/AcOEt 75:25) to give 4- (chloromethyl) benzoic acid 2-butyl-4-chloro-1- [ [2 '- (1-trityltetrazol-5-yl) [1, 1' -biphenyl]-4-yl]Methyl radical]-1H-imidazole-5-methyl ester (1.56g, yield 73%).

2-butyl-4-chloro-1- [ [2 '- (1-trityltetrazol-5-yl) [1, 1' -biphenyl ] 4- (chloromethyl) benzoic acid]-4-yl]Methyl radical]-1H-imidazole-5-methyl ester (0.807g,. 0.98mmol) in CH₃CN (15ml), under nitrogen, AgNO was added in the dark₃(0.305g, 1.8 mmol). The mixture was stirred at 60 ℃ for 6 hours. The silver salt precipitate is then filtered off, the organic phase is diluted with ACOEt and NaH is used₂PO₄(5%, 2X 10ml) and brine (2X 10ml) and washed with Na₂SO₄Drying and concentrating. The crude product was purified by flash chromatography (n-hexane/AcOEt 75:25) to give 4- (nitrooxymethyl) benzoic acid 2-butyl-4-chloro-1- [ [2 '- (1-trityltetrazol-5-yl) [1, 1' -biphenyl]-4-yl]Methyl radical]-1H-imidazole-5-methyl ester (0.553g, 66%).

Acid hydrolysis of 4- (nitrooxymethyl) benzoic acid 2-butyl-4-chloro-1- [ [2 '- (1-trityltetrazol-5-yl) [1, 1' -biphenyl ] -4-yl ] methyl ] -1H-imidazol-5-methyl ester by the method described in method A gave the title compound 4- (nitrooxymethyl) benzoic acid 2-butyl-4-chloro-1- [ [2 '- (1H-tetrazol-5-yl) [1, 1' -biphenyl ] -4-yl ] methyl ] -1H-imidazol-5-methyl ester.

Example 2

4-Nitropoxybutyric acid 2-butyl-4-chloro-1- [ [2 '- (1H-tetrazol-5-yl) [1, 1' -biphenyl ] -4-yl ] methyl ] -1H-imidazol-5-methyl ester (corresponding to Compound (2))

The compound can be obtained by 4 different synthetic methods:

synthesis ofMethod A

To 2-butyl-4-chloro-1- [ [2 '- (1-trityltetrazol-5-yl) [1, 1' -biphenyl ] at 0 deg.C]-4-yl]Methyl radical]-1H-imidazole-5-methanol (1.7g, 2.6mmol) (from example 1), 4-nitrooxybutyric acid (0.536g, 3.6mmol) and N, N-dimethylaminopyridine (0.05g, 0.4mmol) in CH₂Cl₂(20m]) And THF (6ml) was added slowly Dicyclohexylcarbodiimide (DCC) (0.722g, 3.50mmol) in CH₂Cl₂(5ml) solution, the reaction was stirred at room temperature for 24 h. The dicyclohexylurea formed is then filtered off and the organic phase is concentrated. The crude material was purified by flash chromatography (n-hexane/EtOAc 7:3) to give 4-nitrooxybutyric acid 2-butyl-4-chloro-1- [ [2 '- (1-trityltetrazol-5-yl) [1, 1' -biphenyl [ ]]-4-yl]Methyl radical]-1H-imidazole-5-methyl ester (1.45g, 70%).

4-nitrooxybutyric acid 2-butyl-4-chloro-1- [ [2 '- (1-trityltetrazol-5-yl) [1, 1' -biphenyl ] ester]-4-yl]Methyl radical]-1H-imidazole-5-methyl ester (1.0g, 1.25mmol) in CH₂Cl₂(20ml), HCl was bubbled through the solution for 20 min. The reaction was then concentrated and subjected to flash Chromatography (CH)₂Cl₂Acetone 8:2, then acetone) to give the crude compound as a white foam. Dissolving it in H₂O/CH₃CN, and freeze drying to obtain 4-nitrooxybutyric acid 2-butyl-4-chloro-1- [ [2 '- (1H-tetrazol-5-yl) [1, 1' -biphenyl]-4-yl]Methyl radical]-1H-imidazole-5-methyl ester (0.507g, yield 71%) as a white solid.

¹H-NMR(DMSO-d₆)：7.66(2H，d)；7.57(1H，d)；7.49(1H，d)；7.09(2H，d)；6.95(2H，d)；5.25(2H，s)；4.99(2H，s)；4.49(2H，t)；2.54(2H，t)；2.01(2H，t)；1.60(2H，m)；1.49(2H，m)；1.32(4H，m)；0.84(3H，t).

Synthesis method B

To 2-butyl-4-chloro-1- [ [2 '- (1-trityltetrazol-5-yl) [1, 1' -biphenyl ] at 0 deg.C]-4-yl]Methyl radical]-1H-imidazole-5-methanol (from example 1) (1.7g, 2.6mmol), 4-bromobutyric acid (0.561g, 3.36mmol) and N, N-dimethylaminopyridine (0.05g, 0.4mmol) in CH₂Cl₂(20ml) and THF (6ml) solution dicyclohexylcarbodiimide (0.722g, 3.50mmol) in CH was added slowly₂Cl₂(5ml) solution, the reaction was stirred at room temperature for 24 h. The dicyclohexylurea formed is then filtered off and the organic phase is concentrated. Purifying the crude material by silica gel chromatography (n-hexane/ETOAc 75:25) to obtain 4-bromobutyric acid 2-butyl-4-chloro-1- [ [2 '- (1-trityltetrazol-5-yl) [1, 1' -biphenyl]-4-yl]Methyl radical]-1H-imidazole-5-methyl ester (1.27g, yield 60%).

4-bromobutyric acid 2-butyl-4-chloro-1- [ [2 '- (1-trityltetrazol-5-yl) [1, 1' -biphenyl]-4-yl]Methyl radical]-1H-imidazole-5-methyl ester (1.2g, 1.47mmol) in CH₃CN (20ml), under nitrogen, AgNO was added in the dark₃(0.475g, 2.8 mmol). The mixture was stirred at 60 ℃ for 8 hours. It was then partitioned between EtOAc and phosphate buffer (pH 3, 40 ml). The organic phase was washed with phosphate buffer (pH 3, 2 × 25ml), brine (3 × 25ml) and Na₂SO₄Drying and concentrating. The crude material was purified by flash chromatography (n-hexane/AcOEt 7:3) to give 4-nitrooxybutyric acid 2-butyl-4-chloro-1- [ [2 '- (1-trityltetrazol-5-yl) [1, 1' -biphenyl]-4-yl]Methyl radical]-1H-imidazole-5-methyl ester (0.819g, yield 70%) as a foam.

Acid hydrolysis of 4-nitrooxybutyric acid 2-butyl-4-chloro-1- [ [2 '- (1-trityltetrazol-5-yl) [1, 1' -biphenyl ] -4-yl ] methyl ] -1H-imidazol-5-methyl ester by the method described in example 2, method A gave the title compound 4-nitrooxybutyric acid 2-butyl-4-chloro-1- [ [2 '- (1H-tetrazol-5-yl) [1, 1' -biphenyl ] -4-yl ] methyl ] -1H-imidazol-5-methyl ester (0.507g, 71%).

Synthesis method C

To 2-butyl-4-chloro-1- [ [2 '- (1H-tetrazol-5-yl) [1, 1' -biphenyl ] at 0 ℃ under nitrogen]-4-yl]Methyl radical]To a solution of (3.6g, 8.5mmol) of (E) -1H-imidazole-5-methanol, N-dimethylaminopyridine (0.1g, 0.85mmol) and TEA (1.18ml, 0.85mmol) in THF (60ml) was slowly addedA solution of 4-bromobutyryl chloride (0.98ml, 8.5mmol) in THF (1ml) was added and the reaction was stirred at room temperature for 1.5 h. It was then partitioned between EtOAc and phosphate buffer (pH 3, 40ml) and extracted with EtOAc (3 × 15 ml). The organic phase is passed through Na₂SO₄Drying and concentrating. Subjecting the crude material to flash Chromatography (CH)₂Cl₂Acetone 8:2) to obtain 4-bromobutyric acid 2-butyl-4-chloro-1- [ [2 '- (1H-tetrazol-5-yl) [1, 1' -biphenyl]-4-yl]Methyl radical]-1H-imidazole-5-methyl ester (2.5g, yield 51%) as a white solid.

4-bromobutyric acid 2-butyl-4-chloro-1- [ [2 '- (1H-tetrazol-5-yl) [1, 1' -biphenyl]-4-yl]Methyl radical]-1H-imidazole-5-methyl ester (0.56g, 0.98mmol) in CH₃CN (15ml), under nitrogen, AgNO was added in the dark₃(0.83g, 4.9 mmol). The mixture was stirred at 60 ℃ for 8 hours. Then, it was cooled and poured into a phosphate buffer (pH 3, 40 ml). NaCl solid was added and the mixture was extracted with EtOAc. The organic phase was washed with phosphate buffer (pH 3, 2 × 25ml), brine (3 × 25ml) and Na₂SO₄Drying and concentrating. Subjecting the crude material to flash Chromatography (CH)₂Cl₂Acetone 8:2, then acetone) to give the crude compound as a white foam. Dissolving it in H₂O/CH₃CN, and freeze drying to obtain 4-nitrooxybutyric acid 2-butyl-4-chloro-1- [ [2 '- (1H-tetrazol-5-yl) [1, 1' -biphenyl]-4-yl]Methyl radical]-1H-imidazole-5-methyl ester (0.3g, yield 55%) as a white solid.

Synthesis method D

To CH 4-bromobutyric acid (0.91g, 5.4mmol), pentafluorophenol (1.00g, 5.4mmol) and DMAP (0.13g, 1.1mmol) under nitrogen at 0 deg.C₂Cl₂To the solution (10ml) was added N, N-dicyclohexylcarbodiimide (1.70g, 8.1mmol) in portions. After 1h, the reaction was slowly warmed to room temperature and stirred for 5 h. Dicyclohexylurea was filtered off, the mother liquor was concentrated and purified by flash chromatography (n-hexane/EtOAc 98:2) to give 4-bromobutyric acid pentafluorophenyl ester as a colorless oil (1.40g, 78%).

In the microwaveIn (1, 9) reacting pentafluorophenyl 4-bromobutyrate (0.65 g) with AgNO₃(0.83g, 4.9mmol) in CH₃The mixture in CN (8ml) was heated at 70 ℃ for 20 minutes. The salt formed was filtered off, the solvent was concentrated and the residue was purified by flash chromatography (n-hexane/EtOAc 95:5) to give 4-nitrooxybutyric acid pentafluorophenyl ester as a clear oil (0.38g, 62%).

To 2-butyl-4-chloro-1- [ [2 '- (1H-tetrazol-5-yl) [1, 1' -biphenyl ] at 0 deg.C]-4-yl]Methyl radical]To a solution of-1H-imidazole-5-methanol (0.48g, 1.1mmol), TEA (0.16ml, 1.1mmol) and DMAP (0.14mg, 1.1mmol) in DMF (3ml) was added a solution of 4-nitrooxybutyric pentafluorophenyl ester (0.36g, 1.1mmol) in DMF (3 ml). The reaction was slowly warmed to room temperature and stirred for 3 hours. The solvent was then evaporated under reduced pressure. The residue was dissolved in EtOAc (10ml) and washed sequentially with buffer (pH 3) and brine. The organic layer was washed with Na₂SO₄Drying, concentrating, and performing flash Chromatography (CH)₂Cl₂MeOH 98:2) to yield the title compound (0.41g, 66%).

Example 3

11-Nitropoxy undecanoic acid 2-butyl-4-chloro-1- [ [2 '- (1H-tetrazol-5-yl) [1, 1' -biphenyl ] -4-yl ] methyl ] -1H-imidazol-5-methyl ester (corresponding to Compound (68))

Using method A as described in example 2, but starting from 2-butyl-4-chloro-1- [ [2 '- (1-trityltetrazol-5-yl) [1, 1' -biphenyl ] -4-yl ] methyl ] -1H-imidazole-5-methanol (1.7g, 2.6mmol) and 11-nitrooxydecanoic acid (0.78g, 3.36mmol), we obtained 11-nitrooxydecanoic acid 2-butyl-4-chloro-1- [ [2 '- (1-trityltetrazol-5-yl) [1, 1' -biphenyl ] -4-yl ] methyl ] -1H-imidazole-5-methyl ester (1.65g, 80%).

This compound (1.6g, 2.0mmol) was hydrolyzed with acid and Et₂Crystallization of O/n-hexane gives 11-nitrooxydiarylic acid 2-butyl-4-chloro-1- [ [2 '- (1-trityltetrazol-5-yl) [1, 1' -biphenyl]-4-yl]Methyl radical]-1H-imidazole-5-methyl ester (0.91g, 70%).

(DMSO)：7.66(2H，d)；7.57(1H，d)；7.59(1H，d)；7.09(2H，d)；6.95(2H，d)；5.25(2H，s)；4.99(2H，s)；4.49(2H，t)；2.54(2H，t)；2.01(2H，t)；1.62(2H，m)；1.49(2H，m)；1.35-1.14(16H，m)；0.84(3H，t).

Example 4

3- (Nitropoxymethyl) benzoic acid 2-butyl-4-chloro-1- [ [2 '- (1H-tetrazol-5-yl) [1, 1' -biphenyl ] -4-yl ] methyl ] -1H-imidazol-5-methyl ester (corresponding to Compound (5))

2-butyl-4-chloro-1- [ [2 '- (1-trityl-tetrazol-5-yl) [1, 1' -biphenyl ] is used as the raw material]-4-yl]Methyl radical]-1H-imidazole-5-methanol (prepared from example 1) (1.0g, 1.5mmol), triethylamine (0.42ml, 3.0mmol) and N, N-dimethylaminopyridine (36mg, 0.30mmol) were dissolved in CH₂Cl₂(10 ml). 3- (chloromethyl) benzoyl chloride (0.24ml, 1.7mmol) was then added and the reaction stirred at room temperature for 4 h. The mixture was diluted with EtOAC (50ml) and the organic phase was washed with NaH₂PO₄(5％，2×25ml)、NaHCO₃(5%, 2X 25ml), brine (2X 25ml) and Na₂SO₄Drying and concentrating. The crude material was purified by flash chromatography (n-hexane/EtOAC 75:25) to give 3- (chloromethyl) benzoic acid 2-butyl-4-chloro-1- [ [2 '- (1-trityl-tetrazol-5-yl) [1, 1' -biphenyl]-4-yl]Methyl radical]-1H-imidazole-5-methyl ester (1.0g, 81%) as an oil.

2-butyl-4-chloro-1- [ [2 '- (1-trityl-tetrazol-5-yl) [1, 1' -biphenyl ] 3- (chloromethyl) benzoic acid]-4-yl]Methyl radical]-1H-imidazole-5-methyl ester (0.66g, 0.20mmol) was suspended in CH₃CN (10ml), NaI (0.24g, 1.6mmol) was added. The reaction was refluxed for 1 hour and then diluted with EtOAc (25 mL). H for organic phase₂O (3X 25ml) and NaSO₄Drying and concentrating. Dissolving the crude material in CH₃CN (4ml), AgNO was added under nitrogen in the dark₃(0.34g, 2 mmol). The reaction was stirred at room temperature for 2 hours and then diluted with EtOAC (10 ml). NaH for organic phase₂PO₄(5%, 2X 10ml), brine (2X 10ml) and washed with NaSO₄DryingAnd (4) concentrating. The crude material was purified by flash chromatography (hexanes/EtOAc 75:25) to give 2-butyl-4-chloro-1- [ [2 '- (1-trityl-tetrazol-5-yl) [1, 1' -biphenyl ] 3- (nitrooxymethyl) benzoic acid]-4-yl]Methyl radical]-1H-imidazole-5-methyl ester (230mg, 33%).

2-butyl-4-chloro-1- [ [2 '- (1-trityl-tetrazol-5-yl) [1, 1' -biphenyl ] 3- (nitrooxymethyl) benzoic acid]-4-yl]Methyl radical]-1H-imidazole-5-methyl ester (O.23g, 0.27mmol) in CH₂Cl₂(5ml), HCl was bubbled through the solution. After 10min, the reaction was concentrated and flash Chromatographed (CH)₂Cl₂Acetone 8:2, then acetone). The resulting yellow foam was treated with decolorizing carbon and dissolved in H₂O/CH₃CN, and freeze drying to obtain the m-nitrobenzylbenzoic acid 2-butyl-4-chloro-1- [ [2 '- (1H-tetrazol-5-yl) [1, 1' -biphenyl]-4-yl]Methyl radical]-1H-imidazole-5-methyl ester as a white solid (0.11g, 63%).

(CDCl₃)：7.90(2H，m)；7.78(1H，d)；7.56(3H，m)；7.40(1H，m)；7.19(1H，d)；7.06(2H，d)；6.83(2H，d)；5.40(2H，s)；5.24(2H，s)；5.14(2H，s)；2.47(2H，t)；1.61(2H，m)；1.32(2H，m)；0.87(3H，m).

Example 5

6-Nitropoxyhexanoic acid 2-butyl-4-chloro-1- [ [2 '- (1H-tetrazol-5-yl) [1, 1' -biphenyl ] -4-yl ] methyl ] -1H-imidazol-5-methyl ester (corresponding to compound (69)

2-butyl-4-chloro-1- [ [2 '- (1-trityl-tetrazol-5-yl) [1, 1' -biphenyl ] is used as the raw material]-4-yl]Methyl radical]-1H-imidazole-5-methanol (prepared from example 1) (2.0g, 3.0mmol), 6-bromohexanoic acid (0.90g, 4.6mmol), N-dimethylaminopyridine (38mg, 03mmol), triethylamine (1.3ml, 9.3mmol) in CH₂Cl₂(20ml), the solution was cooled to 0 ℃. 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (EDAC) (0.94g, 9.3mmol) was then added and the reaction was slowly warmed to room temperature and stirred overnight. NaH for organic phase₂PO₄(5%, 20ml), brine (20ml)Washing with Na₂SO₄Drying and purification by flash chromatography (n-hexane/EtOAc 7:3) to give 2-butyl-4-chloro-1- [ [2 '- (1-trityl-tetrazol-5-yl) [1, 1' -biphenyl ] 6-bromohexanoic acid]-4-yl]Methyl radical]-1H-imidazole-5-methyl ester as an oil (1.94g, 76%).

2-butyl-4-chloro-1- [ [2 '- (1-trityl-tetrazol-5-yl) [1, 1' -biphenyl ] 6-bromohexanoic acid]-4-yl]Methyl radical]-1H-imidazole-5-methyl ester (0.77g, 0.90mmol) and NaI (0.30g, 2.0mmol) in CH₃CN (10ml), the mixture was refluxed for 1 hour. Then diluted with EtOAc (50mL) and the organic phase was taken with H₂O (2X 25ml) and Na₂SO₄Drying and concentrating. The crude product was suspended in CH₃CN (7ml), AgNO was added₃(0.60g, 3.5 mmol). The reaction was stirred at room temperature under nitrogen in the dark for 3 hours. It was then partitioned between EtOAc (30ml) and phosphate buffer (pH 3, 25 ml). The organic phase was washed with phosphate buffer (pH 3, 2 × 25ml) and brine (3 × 25ml) and washed with Na₂SO₄Drying and concentrating. The crude material was purified by flash chromatography (n-hexane/EtOAc 7:3) to give 6-nitrooxycaproic acid 2-butyl-4-chloro-1- [ [2 '- (1-trityl-tetrazol-5-yl) [1, 1' -biphenyl]-4-yl]Methyl radical]-1H-imidazole-5-methyl ester as foam (0.69g, 64%).

2-butyl-4-chloro-1- [ [2 '- (1-trityl-tetrazol-5-yl) [1, 1' -biphenyl ] 6-nitrooxycaproic acid]-4-yl]Methyl radical]-1H-imidazole-5-methyl ester (0.88g) in CH₂Cl₂(20ml) HCl was bubbled through the solution for 20 minutes. The mixture was then concentrated and subjected to flash Chromatography (CH)₂Cl₂Acetone 8:2, then acetone) to give the product as a yellow foam. Treating with decolorizing carbon, dissolving in H₂O/CH₃CN, and freeze drying to obtain the product 6-nitrooxyhexanoic acid 2-butyl-4-chloro-1- [ [2 '- (1H-tetrazole-5-yl) [1, 1' -biphenyl ]]-4-yl]Methyl radical]-1H-imidazole-5-methyl ester as a white solid (0.41g, 68%).

(CDCl₃)：7.79(1H，d)；7.63-7.49(2H，m)；7.41(1H，d)；7.08(2H，d)；6.77(2H，d)；5.14(2H，s)；4.88(2H，s)；4.38(2H，t)；2.38(2H，t)；2.06(2H，m)；1.70-1.50(6H，m)；1.37-1.30(4H，m)；0.85(3H，t)，

Example 6

2-butyl-4-chloro-1- [ [2 '- (1H-tetrazol-5-yl) [1, 1' -biphenyl ] -4-yl ] methyl ] -1H-imidazole-5-carboxylic acid (3-nitrooxy) propyl ester (corresponding to compound (7))

To a solution of 2-butyl-4-chloro-5-formylimidazole (1.2g, 6.4mmol) in t-ButOH (35ml) and 5% Na₂HPO₄KMnO was added to an aqueous solution (25ml)₄(6.1g, 38.6mmol) of water (40 ml). After 6 minutes at room temperature, 40% NaHSO was added₃The aqueous solution quenches the mixture. The suspension is filtered and washed with H₂O washing, and freeze drying the filtrate. Dissolving the residue in H₂O (50ml), acidified to pH 2.5 with 3N HCl and extracted with EtOAc (3X 70 ml). Combining the organic extracts, passing over Na₂SO₄Drying and evaporation to dryness gave 2-butyl-4-chloro-imidazole-5-carboxylic acid (1.07g, 83%) as a white solid.

Dicyclohexylcarbodiimide (0.91g, 4.4mmol) was added slowly in portions to a solution of 2-butyl-4-chloro-imidazole-5-carboxylic acid (0.61g, 3mmol), 3-bromopropanol (0.52g, 3.74mmol) and N, N-dimethylaminopyridine (0.08g, 0.65mmol) in THF (12ml) at 0 deg.C and the reaction stirred at room temperature for 4 hours. The dicyclohexylurea formed is then filtered off and the organic phase is concentrated. The crude material was purified by silica gel chromatography (n-hexane/AcOEt 8:2) to give 3-bromopropyl 2-butyl-4-chloro-imidazole-5-carboxylate (0.5g, 50%) as a white foam.

2-butyl-4-chloro-imidazole-5-carboxylic acid 3-bromopropyl ester (0.807g, 2.47mmol) was dissolved in CH₃CN (15ml), AgNO was added₃(0.63g, 3.7 mmol). The mixture was stirred at room temperature for 8 h. The silver salt precipitate is then filtered off, the organic phase is diluted with ACOEt and NaH is used₂PO₄(5%, 2X 10ml) and brine (2X 10ml) and washed with Na₂SO₄Drying and concentrating. The crude material was purified by flash chromatography (n-hexane/AcOEt 70:30) to give 2-butyl-4-chloro-imidazole-5-carboxylic acid 3-nitrooxypropyl ester (0.377g, 50%).

To a solution of 3-nitrooxypropyl 2-butyl-4-chloro-imidazole-5-carboxylate (0.76g, 2.5mmol) in Dimethylacetamide (DMA) (13ml) was slowly added potassium tert-butoxide (0.28g, 2.5mmol) in portions at 0 ℃ under nitrogen. After stirring for 10min, a solution of N- (trityl) -5- (4' -bromomethylbiphenyl-2-yl-) tetrazole (1.7g, 3mmol) in DMA (10ml) was added and the mixture stirred at room temperature for 1 h. The mixture was then partitioned between water and EtOAc. Separating the organic phase over Na₂SO₄Drying and concentrating. The crude material was purified by flash chromatography (n-hexane/EtOAc 7:3) to afford 2-butyl-4-chloro-1- [ [2 '- (1-trityltetrazol-5-yl) [1, 1' -biphenyl]-4-yl]Methyl radical]-1H-imidazole-5-carboxylic acid 3-nitrooxypropyl ester (1.56g, 80%).

Acid hydrolysis of 3-nitrooxypropyl 2-butyl-4-chloro-1- [ [2 '- (1-trityltetrazol-5-yl) [1, 1' -biphenyl ] -4-yl ] methyl ] -1H-imidazole-5-carboxylate (1g, 1.28mmol) by a method analogous to that described for method A in example 1, to give the title compound (white solid) (0.28g, 40%).

¹H-NMR(DMSO-d₆)：7.60-7.20(4H，m)；7.12(2H，d)；6.92(2H，d)；5.72(2H，s)；4.58(2H，t)；4.50(2H，t)；2.54(2H，t)；2.31(2H，m)；1.49(2H，m)；1.32(2H，m)；0.84(3H，t).

Investigation of vascular tone (tone)

To compare the ability of the nitro derivative of ARB to induce vasodilation with the parent ARB, ex vivo rabbit thoracic aortic preparations (preparations) were tested in vitro (Wanstall J.C. et al, Br.J.Pharmacol., 134: 463-472, 2001). Male New Zealand rabbits were anesthetized with thiopentan sodium (50mg/kg, iv), sacrificed by exsanguination, and then the aorta was excised by thoracotomy. Aortic ring specimens (4mm long) were prepared in small organ chambers (5ml) in Physiological Saline Solution (PSS) at 37 ℃. The composition of PSS is (mM): NaCl 130, NaHCO₃ 14.9，KH₂PO₄ 1.2，MgSO₄ 1.2，HEPES10，CaCl₂Ascorbic acid 170 and glucose 1.1 (95% O)₂/5％CO₂(ii) a pH 7.4). Each ring was subjected to 2g of passive tension. Isometric tension was recorded with a Grass sensor (Grass FT03) connected to a BIOPAC MP150 system. The specimen was allowed to equilibrate for 1h, then the next largest contraction was given with norepinephrine (NA, 1 μ M), and after the contraction stabilized, acetylcholine (ACh, 10 μ M) was added. The relaxation response to ACh indicates the presence of functional endothelium. Vessels that failed to contract NA or did not relax in Ach stimulation were discarded. When stable presystraction was achieved, a cumulative concentration-response curve for either vascular relaxant was obtained in the presence of functional endothelium. Each arterial loop was exposed to only one combination of inhibitor and vascular relaxation agent. In addition, the effect of the compound on vascular relaxation caused by the soluble guanylate cyclase inhibitor ODQ (1-H- (1, 2, 4) -oxadiazole (4, 3-a) quinoxalin-1-one) was examined by preincubating the aortic ring with ODQ (10. mu.M) for 20 min.

The response to the relaxant is expressed as a percentage of residual shrinkage and plotted against the concentration of the test compound. By interpolation, the IC is obtained from these figures₅₀Value (wherein IC)_5oIs the concentration of test compound that produces 50% of the maximum relaxation).

During the experiment, in these aortic rings, plateau values obtained with NA were stable, with no significant spontaneous loss of contraction. Under these experimental conditions, any tested concentration of ARB losartan did not produce a relaxation effect, and the curve was not different from that established in the presence of the vehicle alone.

As shown in Table 1, the nitro derivatives of the present invention produced relaxation in a concentration-dependent manner. Furthermore, in experiments performed in the presence of ODQ (10 μ M), the vascular relaxation response to the test compound was inhibited.

TABLE 1

Compound (I)	IC(μM)±sem
		Losartan	Up to 100. mu.M no effect
Example 1 Compounds	33±12
		Example 2 Compounds	15±3
Example 4 Compounds	54±16
		Example 5 Compounds	18±6

IC₅₀Is the concentration inhibiting 50% of the reaction

Effect of losartan Nitro derivatives on inflammatory pathways in vitro

The experiment was performed using the RAW 264.7 monocyte macrophage cell line. Cells were stimulated for 16h in the presence of Lipopolysaccharide (LPS) (1. mu.g/ml). At the end of the incubation, the medium was collected and analyzed for nitrite content using a standard Griess reaction.

The results are shown in table 2, expressed as% nitrite content per treatment relative to LPS treated samples.

TABLE 2

As shown in table 2, unlike the parent compound, the nitro derivative (compound of example 4) inhibited nitrite accumulation by LPS.

Research on in vitro anti-platelet activity of losartan nitro derivative

The ability of losartan nitro derivatives to inhibit human platelet aggregation in vitro was evaluated. Platelet aggregation was measured according to the Born method (Gresele P, Arnout J, Deckmyn H et al, J Clin invest.1987; 80: 1435-45) using 0.25ml of a platelet-action (reach) plasma (PRP) sampleAnd (4) collecting. Based on the fact that this class of agonists is sensitive to the action of nitric oxide, TxA is used₂Analog U46619 as a coagulant. The compound was incubated at 37 ℃ for 2min, and then the clotting agent was added. Aggregation was performed for 5min and the maximum amplitude (cm) was measured. DMSO (0.05% final concentration) was used as vehicle. The concentration of test compound was 10-100. mu.M.

TABLE 3

As shown in table 3, the nitro derivatives were able to significantly inhibit platelet aggregation caused by U46619. Losartan showed a weak effect.

Research on in vivo antihypertensive activity of losartan nitro derivative

The ability of the losartan nitro derivative (compound of example 2) to lower blood pressure was evaluated in conscious Spontaneously Hypertensive Rats (SHRs). Two groups of SHRs (250-300g) were given daily oral doses of losartan (10mg/kg po) or losartan nitro derivative (equimolar dose) for 3 consecutive days. After administration, Systolic Blood Pressure (SBP) and heart rate were monitored at different time points by teleassays.

TABLE 4

As shown in table 4, the nitro derivative (compound of example 2) was able to produce significant reductions in blood pressure levels over the treatment period, unlike the parent compound.

Claims (14)

1. A compound of general formula (I) or a pharmaceutically acceptable salt or stereoisomer thereof:

R-(Y-ONO₂)_s (I)

wherein:

s is an integer equal to 1 or 2;

r is selected from the group consisting of angiotensin II receptor blocker residues of the following formula (II) or (III):

wherein:

R₀is composed of

Or a group-N capable of binding to Y₀Which has one of the following meanings:

-COO-, -O-, -CONH-, -OCO-, -OCOO-or

Wherein R 'and R' are the same or different and are H or straight or branched C₁-C₄An alkyl group;

R₁selected from:

or

Wherein m is an integer equal to 0 or 1, and N₀The definition is the same as above;

wherein N is₁Meaning of (A) and N₀Same or equal to-COOH; provided that at least one N₁The group is equal to-COO-or-CONH-, i.e. it is a group capable of binding to Y;

y is a divalent group having the following meaning:

a)

-straight or branched C₁-C₂₀Alkylene, preferably C₁-C₁₀Optionally substituted with one or more substituents selected from: halogen atom, hydroxy group, -ONO₂Or T₀Wherein T is₀is-OC (O) (C)₁-C₁₀Alkyl) -ONO₂or-O (C)₁-C₁₀Alkyl) -ONO₂；

Cycloalkylene having 5 to 7 carbon atoms in the cycloalkylene ring, said ring being optionally substituted with a side chain T, wherein T is a linear or branched alkyl group having 1 to 10 carbon atoms, preferably CH₃；

b)

c)

Wherein n is an integer of 0 to 20, and n¹Is an integer of 1 to 20;

d)

wherein:

n¹is as defined above, and n²Is an integer of 0 to 2;

X₁is-OCO-or-COO-, and R²Is H or CH₃；

e)

Wherein:

n¹、n²、R²and X₁The definition is the same as above;

Y¹is-CH₂-CH₂-or-CH ═ CH- (CH)₂)_n ²-；

f)

Wherein:

n¹and R²As defined above, R³Is H or-COCH₃；

Provided that-ONO is present when Y is selected from the divalent radicals mentioned under b) -f)₂Group and- (CH)₂)_n ¹Group attachment;

g)

wherein X₂is-O-or-S-, n³Is an integer from 1 to 6, preferably from 1 to 4, R²The definition is the same as above;

h)

wherein:

n⁴is an integer of 0 to 10;

n⁵is an integer of 1 to 10;

R⁴、R⁵、R⁶、R⁷are identical or different and are H or straight-chain or branched C₁-C₄Alkyl, preferably R⁴、R⁵、R⁶、R⁷Is H;

wherein-ONO₂The radical being attached to

Wherein n is⁵The definition is the same as above;

Y²is a saturated, unsaturated or aromatic 5-or 6-membered heterocyclic ring containing one or more heteroatoms selected from nitrogen, oxygen, sulphur,

and is selected from

2. A compound of general formula (I) according to claim 1, or a pharmaceutically acceptable salt or a stereoisomer thereof, wherein Y is a divalent group having the following meaning:

a) straight or branched C₁-C₁₀Alkylene, optionally substituted by T₀Is substituted, wherein T₀The definition is the same as above;

b)

wherein n is an integer equal to 0 or 1, and n¹Is an integer equal to 1; provided that-ONO₂Group and- (CH)₂)_n ¹Group attachment;

g)

wherein X₂is-O-or-S-, n³Is an integer equal to 1, and R²Is H.

3. A compound according to claims 1-2 selected from

4. A compound of general formula (I) according to claims 1-3 for use as a medicament.

5. Use of a compound according to claims 1-3 for the manufacture of a medicament having anti-inflammatory, antithrombotic and antiplatelet activity.

6. Use of a compound according to claims 1-3 for the preparation of a medicament useful for the treatment or prevention of cardiovascular diseases, renal diseases and chronic liver diseases, inflammation and metabolic syndrome.

7. Use of a compound of claim 6 in the manufacture of a medicament useful for treating or preventing: heart failure, myocardial infarction, ischemic stroke, atherosclerosis, ocular hypertension and pulmonary hypertension, diabetic nephropathy, peripheral vascular disease, left ventricular dysfunction and hypertrophy, liver fibrosis and portal hypertension.

8. A pharmaceutical composition comprising a pharmaceutically acceptable carrier and a pharmaceutically effective amount of a compound of general formula (I) according to claims 1-3 or a salt or stereoisomer thereof.

9. The pharmaceutical composition of claim 8, which is administered by inhalation spray or aerosol or iontophoresis device in a form suitable for oral, parenteral, rectal, topical and transdermal administration.

10. A liquid or solid pharmaceutical composition in the form of optionally enteric coated tablets, capsules and pills, powders, granules, gels, emulsions, solutions, suspensions, syrups, elixirs, injections, suppositories, transdermal patches or liposomes for oral, parenteral, rectal, topical and transdermal administration or for administration by inhalation, which comprises a compound of formula (I) or a salt or stereoisomer thereof according to claims 1 to 3 and a pharmaceutically acceptable carrier.

11. A pharmaceutical composition comprising a compound of formula (I), at least one compound useful for treating cardiovascular disease, and a pharmaceutically acceptable carrier.

12. The pharmaceutical composition of claim 11, wherein the compound for treating cardiovascular disease is selected from the group consisting of: ACE inhibitors, HMGCoA reductase inhibitors, beta-adrenergic blockers, calcium channel blockers, diuretics, antithrombotic agents such as aspirin, nitrated ACE inhibitors, nitrated HMGCoA reductase inhibitors, nitrated beta-adrenergic blockers, nitrated aspirin and nitrated diuretics.

13. A kit comprising a compound of general formula (I) as defined in claim 1, a compound for use in the treatment of cardiovascular diseases, for simultaneous, separate or sequential use as a combined preparation in the treatment of cardiovascular diseases.

14. The kit of claim 13, wherein said compound for treating cardiovascular disease is selected from the group consisting of: ACE inhibitors, HMGCoA reductase inhibitors, beta-adrenergic blockers, calcium channel blockers, diuretics, antithrombotic agents such as aspirin, nitrated ACE inhibitors, nitrated HMGCoA reductase inhibitors, nitrated beta-adrenergic blockers, nitrated aspirin and nitrated diuretics.