CZ20002556A3 - Hypolipidemic benzothiazepine compounds - Google Patents

Abstract

)Annotation: The solution relates to new hypolipidemic compounds of general formula I, where R1 is H or methyl, and their salts, solvates or physiologically functional derivatives, processes and novel intermediates for their preparation, pharmaceutical compositions containing them, and their use in medicine, especially in the prophylaxis and treatment of hyperlipidemic conditions and associated conditions, such as atherosclerosis.

Description

Hypolipidemic benzothiazepine compounds

Technical field

The invention relates to novel hypolipidemic compounds, processes and novel intermediates for their preparation, pharmaceutical compositions containing them and their use in medicine, in particular in the prophylaxis and treatment of hyperlipidemic conditions and associated conditions such as atherosclerosis.

State of the art

Hyperlipidemic states are often associated with increased plasma concentrations of low-density lipoprotein cholesterol, LDL. These concentrations can be reduced by reducing the absorption of bile acids from the intestine. One way to achieve this is to inhibit the active uptake of bile acids in the terminal part of the small intestine. This inhibition promotes the conversion of cholesterol to bile acids in the liver, resulting in an increased need for cholesterol and thus increased removal of LDL-cholesterol from the blood plasma or serum.

The compounds of the present invention reduce the concentration of LDL cholesterol in plasma or blood serum and are therefore particularly useful as hypolipidemic agents. By reducing the concentration of cholesterol and its esters in plasma, the compounds of the invention slow down the formation of sclerotic lesions and thereby reduce the incidence of symptoms associated with coronary heart disease. The incidence of cardiac events is dependent on the increased concentration of cholesterol and its esters in plasma or serum.

International Patent Application No. PCT/GB/0300328 describes 1,4-benzothiazepine compounds which have hypolipidemic activity. International Patent Application No. PCT/GB/95/02700 (published as WO/9616051) describes 1,5-benzothiazepine compounds which also have hypolipidemic activity. A group of substituted 1,5-benzothiazepine compounds have been discovered which have surprising hypolipidemic activity over the compounds described in the prior art.

• · · ···· ····

The essence of the invention

The present invention provides a compound of general formula I

where

R ¹ is H or methyl; or a salt, solvate or physiologically functional derivative thereof.

Preferably, R ¹ is hydrogen.

Suitable compounds of general formula I are selected from the group consisting of:

(±)-2,3,4,5-Tetrahydro-3-ethyl-3-butyl-5-phenyl-7-chloro-8-hydroxy-1,5-benzothiazepine 1,1-dioxide;

(3S)-2,3,4,5-Tetrahydro-3-ethyl-3-butyl-5-phenyl-7-chloro-8-hydroxy-1,5-benzothiazepine-1,1-dioxide; and (±)-2,3,4,5-Tetrahydro-3-ethyl-3-butyl-5-phenyl-7-chloro-8-methoxy-1,5-benzothiazepine-1,1-dioxide; or a salt, solvate or physiologically functional derivative thereof.

Pharmaceutically acceptable salts are particularly suitable for medical use because they are more soluble in water than the parent, i.e. basic, compounds. Such salts must have a pharmaceutically acceptable anion or cation. Suitable pharmaceutically acceptable acid addition salts of the compounds of the present invention include salts derived from inorganic acids such as hydrochloric, hydrobromic, phosphoric, metaphosphoric, nitric, sulfonic and sulfuric acids, and from organic acids such as acetic, benzenesulfonic, benzoic, citric, ethanesulfonic, fumaric, gluconic, glycolic, isothionic, lactic, lactobionic, maleic, malic, methanesulfonic, succinic, p-toluenesulfonic, tartaric and trifluoroacetic acids. The chloride is particularly preferred for medical use. Suitable pharmaceutically acceptable basic salts include ammonium salts, alkali metal salts such as sodium and potassium salts, and alkaline earth element salts such as magnesium and calcium salts.

Salts with an anion that is not pharmaceutically acceptable are subject to the invention as useful intermediates for the preparation or purification of pharmaceutically acceptable salts and/or for use in non-therapeutic applications, for example in vitro.

Any reference to "compounds of formula I", "compounds of the present invention", "compounds of the invention", etc. refers to compounds of formula I as described above or salts, solvates or physiologically functional derivatives thereof.

The term "physiologically functional derivative" refers herein to any physiologically acceptable derivative of a compound of the present invention, for example an ester, which upon administration to a mammal, such as a human, is capable of providing (directly or indirectly) such compound or an active metabolite thereof. Such derivatives are known to those skilled in the art and are referenced in Burger's Medicinal Chemistry And Drug Discovery, 5th Edition, Volume 1: Principles And Practice, incorporated herein by reference.

Physiologically functional derivatives which are capable of providing (directly or indirectly) upon administration to a mammal such as a human the compound of the invention or its active metabolite are generally referred to as prodrugs. Prodrugs may or may not be active in themselves. Suitable prodrugs of the present invention are formed at the R ¹ position to form a C 1-6 ester or C 1-6 alkoxy group.

Active metabolites are those metabolites that can be generated in vivo by metabolism of the compounds of the present invention and include, for example, glucuronides.

The compounds of the present invention may also exist in various polymorphic forms, for example, amorphous and crystalline amorphous forms. All polymorphic forms of the compounds of the present invention are within the scope of the invention and are a further aspect thereof.

Compounds of formula I exist in forms where the central carbon atom C(Et)(n-Bu)- is chiral. The present invention includes within its scope each possible optical isomer substantially free, i.e. accompanied by less than 5% of any other optical isomer(s), and mixtures of one or more optical isomers in any proportions, including racemic mixtures. The (S)-isomer is preferred.

According to further aspects of the invention, there are also provided:

(a) compounds of formula I and their pharmaceutically acceptable salts, solvates and physiologically functional derivatives for use as therapeutic agents, especially in the prevention and treatment of clinical conditions for which a bile acid absorption inhibitor is indicated, for example a hyperlipemic condition such as atherosclerosis;

(b) pharmaceutical compositions comprising a compound of formula I or pharmaceutically acceptable salts, solvates or physiologically functional derivatives thereof, at least one pharmaceutically acceptable carrier and optionally one or more other physiologically active agents;

(c) the use of a compound of formula I or a pharmaceutically acceptable salt, solvate or physiologically functional derivative thereof for the manufacture of a composition for the prevention and treatment of clinical conditions for which a bile acid absorption inhibitor is indicated, for example a hyperlipidemic condition such as atherosclerosis;

(d) a method of inhibiting bile acid absorption from the intestine of a mammal, such as a human, comprising administering to the mammal an effective amount of a bile acid absorption inhibitor of a compound of formula I or a pharmaceutically acceptable salt, solvate or physiologically functional derivative thereof;

(e) a method of reducing LDL and VLDL cholesterol concentrations in blood plasma or serum in a mammal, such as a human, comprising administering to the mammal a cholesterol-lowering effective amount of a compound of formula I, or a pharmaceutically acceptable salt, solvate, or physiologically functional derivative thereof;

• · • · · · • · · · • · · · · · ·· ···· « · · • · · · · ·· ····· ·· ·· ·· ·· ·· ·· · (f) a method of reducing the concentrations of cholesterol and cholesterol esters in blood plasma or serum in a mammal, such as a human, comprising administering to the mammal a cholesterol or cholesterol ester-lowering effective amount of a compound of formula I or a pharmaceutically acceptable salt, solvate or physiologically functional derivative thereof;

(g) a method of increasing fecal bile acid excretion in a mammal, such as a human, comprising administering to the mammal an effective amount of a compound of formula I or a pharmaceutically acceptable salt, solvate or physiologically functional derivative thereof;

(h) a method of preventing or treating a clinical condition in a mammal, such as a human, for which a bile acid absorption inhibitor is indicated, for example a hyperlipidemic condition such as atherosclerosis, which comprises administering to the mammal a therapeutically effective amount of a compound of formula I or a pharmaceutically acceptable salt, solvate or physiologically functional derivative thereof;

(i) a method of reducing the incidence of coronary heart disease-related events in a mammal, such as a human, comprising administering to the mammal an effective amount of a compound of formula I, or a pharmaceutically acceptable salt, solvate, or physiologically functional derivative thereof, to the mammal, such as a human, comprising administering to the mammal an effective amount of a compound of formula I, or a pharmaceutically acceptable salt, solvate, or physiologically functional derivative thereof;

(j) a method of reducing the concentration of cholesterol in blood plasma or serum in a mammal, such as a human, comprising administering a cholesterol-lowering effective amount of a compound of formula I;

(k) a process for the preparation of compounds of formula I (including salts, solvates or physiologically functional derivatives as defined herein); and (I) novel chemical intermediates in the preparation of compounds of formula I.

• ··· ····

The compounds of the present invention may be administered in conjunction with other physiologically active agents, including hypolipidemic agents such as bile acid sequestrants, fibric acid derivatives, or HMG-CoA reductase inhibitors (competitive inhibitors of 3-hydroxy-3-methylglutaryl coenzyme A reductase), for example statins such as pravastatin, lovastatin, fluvastatin, or simvastatin.

The amount of compound of formula I required to achieve the desired biological effect will, of course, depend on a number of factors, for example, the particular compound selected, the intended use, the route of administration and the clinical condition of the recipient. In general, the daily dose is in the range of 0.001 mg to 100 mg (typically 0.01 mg to 50 mg) per day per kilogram of body weight, for example 0.01-10 mg/kg/day. An oral dosage unit formulation, such as a tablet or capsule, may contain, for example, from 0.1 mg to 100 mg/kg, preferably 0.1 to 5 mg.

In the case of a pharmaceutically acceptable salt, the weights indicated above are based on the weight of the benzothiazepine ion derived from the salt.

For the prevention or treatment of the above conditions, the compounds of formula I may be used as compounds per se, but are preferably presented with an acceptable carrier in the form of a pharmaceutical composition. The carrier must, of course, be acceptable in the sense of compatibility with the other ingredients of the composition and must not be deleterious to the recipient. The carrier may be solid, liquid or both, and is preferably used with the compound as a unit dose formulation, for example a tablet, which may contain from 0.05 to 95% by weight of the active compound. Other pharmacologically active compounds may also be present, including other compounds of formula I. The pharmaceutical compositions of the invention may be prepared by any of the known pharmaceutical procedures which essentially involve mixing the ingredients.

When a compound of formula I is used in combination with one or more physiologically active agents as described above, the amount of such additional physiologically active agents to achieve the desired physiological effect will also depend on a number of factors. The specific dosage and dosage regimen will be readily determined by one of ordinary skill in the art. Generally, the dosage used will be that approved for use in humans.

The pharmaceutical compositions of the present invention include compositions suitable for oral, rectal, topical, buccal (e.g. sublingual) and parenteral (e.g. subcutaneous, intramuscular, intradermal or intravenous) administration, although the most suitable route in any given case will depend on the nature and severity of the condition being treated and the nature of the particular compound of formula I being used. Also within the scope of the invention are enteric and controlled release enteric formulations. Suitable coatings for enteric formulations include cellulose acetate phthalate, polyvinyl acetate phthalate, hydroxypropyl methylcellulose phthalate and anionic polymers of methacrylic acid and methacrylic acid methyl ester. Suitable enteric and controlled release enteric formulations include tablets and capsules.

Pharmaceutical compositions suitable for oral administration may be presented in discrete units such as capsules, cachets, troches or tablets, each containing a predetermined amount of the compound of formula I; as a powder or granules; as a solution or suspension in an aqueous or non-aqueous liquid; or as an oil-in-water or water-in-oil emulsion. As indicated, such compositions may be prepared by any suitable pharmacological process which includes the step of bringing the active compound into association with the carrier (which may contain one or more accessory ingredients). In general, the compositions are prepared by homogeneously and intimately mixing the active compound with a liquid or finely divided solid carrier or both, and then, if necessary, shaping the product. For example, a tablet may be prepared by compressing or molding a powdered or granular compound, optionally with one or more other ingredients. Compressed tablets may be prepared by compressing the compound in a free-flowing state, such as powder or granules, in a suitable machine, optionally mixed together with a binder, lubricant, inert diluent and/or surfactant/dispersant(s). Molded tablets may be prepared by molding the powdered compound, moistened with an inert liquid diluent, in a suitable machine. Controlled-release tablets are prepared in a similar manner and with the addition of, for example, hydroxypropylmethylcellulose.

Enteric-coated tablets can be prepared by coating the tablets with an enteric polymer such as cellulose acetate phthalate, polyvinyl acetate phthalate, phthalate ···· · ···

hydroxypropylmethylcellulose or anionic polymers of methacrylic acid and its methyl ester (Eudragit L). In addition to Eudragit L, these polymers should also contain 10% by weight (based on the amount of polymer used) of a plasticizer to prevent the membrane from cracking during administration or storage. Suitable plasticizers include diethyl phthalate, tributyl citrate and triacetin.

Enteric-coated controlled-release tablets can be prepared by coating the tablets with an enteric polymer such as cellulose acetate phthalate, polyvinyl acetate phthalate, hydroxypropyl methylcellulose phthalate or anionic polymers of methacrylic acid and its methyl ester (Eudragit L). In addition to Eudragit L, these polymers should contain 10% by weight (based on the amount of polymer used) of a plasticizer to prevent the membrane from cracking during administration or storage. Suitable plasticizers include diethyl phthalate, tributyl citrate and triacetin.

Capsules can be prepared by mixing a compound of formula I with, for example, magnesium stearate, pregelatinized starch, sodium starch glycolate and/or magnesium stearate and filling the resulting mixture into two-piece hard gelatin capsules.

Controlled release capsules can be prepared by mixing a compound of formula I with, for example, microcrystalline cellulose and/or lactose, the mixture is then extruded using an extruder, and the extrudate is then spheronized and dried. The dried pellets are provided with a controlled release coating, for example, of ethyl cellulose, and then filled into two-piece hard gelatin capsules.

Enteric-coated capsules can be prepared by mixing a compound of formula I with, for example, microcrystalline cellulose and/or lactose, the mixture is then extruded using an extruder, and the extrudate is then spheronized and dried. The dried pellets are provided with an enteric-coated membrane, for example, of cellulose acetate phthalate containing a plasticizer, for example, diethyl phthalate, and then filled into two-piece hard gelatin capsules.

Pharmaceutical compositions suitable for buccal (sublingual) administration include lozenges containing a compound of formula I in a flavored base, usually sucrose or tragacanth, and lozenges containing the compound in an inert base, such as gelatin and glycerin or sucrose and acacia.

Pharmaceutical compositions suitable for parenteral administration will generally comprise sterile aqueous compositions of the compound of formula I, preferably isotonic with the blood of the recipient. These compositions are preferably administered intravenously, although administration may also be effected by subcutaneous, intramuscular or intradermal injection. Such compositions may conveniently be prepared by mixing the compound with water and adjusting to a sterile solution, isotonic with the blood. The injectable compositions of the invention will generally contain from 0.1 to 5% by weight of the active compound.

Pharmaceutical compositions suitable for rectal administration are usually in the form of suppositories containing a unit dose. They may be prepared by admixing a compound of formula I with one or more conventional solid carriers, for example cocoa butter, and then shaping the resulting mixture.

Transdermal administration is also possible. Pharmaceutical compositions suitable for transdermal administration may be provided as individual patches adapted to be maintained in contact with the epidermis of the recipient for a prolonged period of time. Such patches preferably contain the active compound in an aqueous solution, optionally buffered, dissolved or dispersed in an adhesive or dispersed in a polymer. A suitable concentration of the active compound is about 1 to 35%, preferably about 3 to 15%. A particular example is the possibility of treatment by absorption of the active substance from the patch by the action of an electric current or iontophoresis, for example according to the publication Pharmaceutical Research, 3(6), 318 (1986).

Compounds of general formula I can be prepared by conventional methods known to those skilled in the art or by analogous procedures, the principle of which is described in the literature.

For example, compounds of general formula I, where R ¹ is H, can be prepared from compounds of general formula II • ·« · ·

where R ^1a is an alkyl moiety (for example C 1-4 alkyl, preferably methyl), by dealkylation with a suitable reagent such as boron tribromide in a suitable organic solvent, for example methylene chloride.

According to the second process (B), a compound of general formula I, wherein R ¹ is methyl, or a salt, solvate or physiologically functional derivative thereof, can be prepared from a compound of general formula III

(III) by oxidation of sulfur, for example with a mixture of osmium tetroxide and N-methylmorpholine-N-oxide.

Compounds of formula II or III can be prepared from compounds of formula IV, where R ^1a has the meaning given above, by methods known in the art, in particular those described in WO96/16051.

(IV) • · » ···» ···· • ·

Compounds of formula IV can be prepared by the methods described in WO96/16051 or by reacting compounds of formula V with compounds of formula Va.

where Z is a suitable leaving group, for example halo, by first reacting a compound of formula V with a base, for example aqueous potassium hydroxide at an elevated temperature, for example 100°C, cooling and then adding a compound of formula Va.

Compounds of general formula V can be prepared from compounds of general formula VI

by reaction with ammonium thiocyanate and bromine in a suitable solvent such as acetic acid.

Compounds of formula VI are commercially available or can be prepared by methods well known to those skilled in the art.

Compounds of general formula Va can be prepared from compounds of general formula VII

^R2O

Et

HOn>Bu (VII) * ·« * 9 9 t 9 · 9 4 V • 4 » · · · » p í ·* · · · * · · · 4 9 *· · · · · * · * · * ·

Φ··4 4··4 ·· >· 9 9 9 9 where R ² is a suitable hydroxy protecting group, for example tert-butyldimethylsilyl, by oxidation of a compound of general formula VII, for example with sodium periodate and ruthenium chloride in a suitable solvent, such as carbon tetrachloride/acetonitrile/water. Following oxidation, the R ² hydroxy protecting group is removed and converted to a suitable leaving group Z in a known manner, for example with HBr.

Compounds of formula VII can be prepared from the corresponding diols by methods well known or available to one skilled in the art. The diols are commercially available or can be prepared by methods known or available to one skilled in the art.

Compounds of formula I, substantially free of other optical isomers, can be prepared by chiral synthesis, for example using appropriate chiral starting materials, for example chiral compounds of formula Va, or by resolution of products obtained by achiral synthesis, for example by chiral HPLC, enzymatic resolution or classical resolution using chiral acids.

Optional conversion of a compound of formula I, optionally containing a basic substituent, into a corresponding acid addition salt can be carried out by reaction with a solution of the appropriate acid, for example with one of the acids mentioned above. Optional conversion of a compound of formula I containing an acidic substituent into a corresponding base salt can be carried out by reaction with a solution of the appropriate base, for example sodium hydroxide. Optional conversion into a physiologically functional derivative, for example an ester, can be carried out by methods known from the literature.

Furthermore, it is possible to convert derivatives of the general formula I into other derivatives of the general formula I by conventional methods, for example by methylation of the hydroxyl group.

In general, in the following examples, the proton magnetic resonance spectrum is always recorded at 300 MHz. Mass spectra were recorded at atmospheric pressure under chemical ionization, APCI, or on an LCMS or were performed by Oneida Research Services, Inc. under chemical ionization conditions of Cl using methane as the reaction gas. Elemental analysis was performed by Atlantic Microlab, Inc. All reactions were carried out under nitrogen. For thin layer chromatography • · » · · · ···· • · · · ····· ·

........ ·* *· ” ” TLC layer Whatman MK6F silica gel 60 plates were used, spots were visualized by UV light. Column chromatography was performed using EM Science silica gel 60 with a particle diameter of 230 to 400 mesh. Unless otherwise stated, Aldrich Chemical Co. reagents were used without further purification. In addition, Aldrich Chemical Co. anhydrous solvents were used.

The following examples are provided to illustrate the invention, but do not limit the scope of the invention in any way.

Examples of embodiments of the invention

Example 1

Preparation of (3S)-2,3,4,5-tetrahydro-3-ethyl-3-butyl-5-phenyl-7-chloro-8-methoxy-1,5benzothiazepine-1,1-dioxide and (3S)-2,3,4,5-tetrahydro-3-ethyl-3-butyl-5-phenyl-7-chloro-8-hydroxy-1,5-benzothiazepine-1,1-dioxide (±)-2-((Tert.butyldimethylsilyl)oxy)methylethylhexanol (1)

To a slurry of 60% NaH (21.2 g) in 800 mL THF was added 2-n-butyl-2ethyl-1,3-propanediol (85.0 g) in 3 portions and the mixture was stirred for 1 hour. The mixture was cooled to 0 °C.

To the resulting gum was added a 1 M solution of tert-butyldimethylsilyl chloride in THF (530 mL) and the mixture was stirred overnight at room temperature, allowing the temperature to reach room temperature. The solvent was evaporated and the residue was partitioned between water (400 mL) and ether (300 mL). The ether layer was washed with bicarbonate solution and brine and concentrated. Column chromatography (5% ethyl acetate/petroleum ether) gave 1 as a colorless oil (142.6 g).

MS Da/e = 275 (MH ⁺ ).

Calcd for _C15H34O2Si : C, _65.63 ; _H , 12.48. Found; C, 65.87; H, 12.47.

(±)-2-(Bromomethyl)-2-ethylhexanoic acid (2)

To a solution of 1 (142.6 g) in 300 mL _CCl4 , 300 mL _CH3CN , and 450 mL _H2O at 0 °C was added _Na1O4 (283 g) and _RuCl3 (2.0 g) and the mixture was stirred for 20 h while the reaction mixture was warmed to room temperature. The reaction mixture was filtered through Celite and the filtrate was concentrated on a rotary evaporator. The residue was transferred to a separatory funnel and partitioned between _H2O and CH2Cl2. The aqueous phase was extracted three times with _CH2Cl2 , _dried , and concentrated. The residue was taken up in 48% HBr (500 mL) and heated at reflux for 24 h. After cooling to room temperature, it was transferred to a separatory funnel, extracted three times with ethyl ether, washed once with brine, dried over Na ₂ SO ₄ and concentrated. The product was purified by column chromatography on silica gel eluting with 20% ethyl acetate/petroleum ether to give 2 (111 g).

MS Da/e = 157 (M - Br), 237, 239 (M + 1).

Calcd for C8H18ChBr; C, 45.59; H, 7.23; Br, 33.70. Found: C, 46.27; H, 7.17; Br, 32.94.

2-Amino-5-chloro-6-methoxybenzothiazole (4)

To a solution of 3-chloro-p-anisidine (3, 58.4 g) in acetic acid (400 ml) was added ammonium thiocyanate and the mixture was stirred for 30 minutes at room temperature. The reaction mixture was cooled to 15 ^°C in an ice bath. To the cooled reaction mixture was added a solution of bromine (17.2 ml) in acetic acid (200 ml) over 10 minutes. After stirring for 10 minutes, the reaction mixture was allowed to warm to room temperature. The mixture was stirred for 3.5 hours, the reaction mixture was filtered and the solid was collected on filter paper. The solid was transferred to a container and 200 ml of water was added. The suspension was stirred vigorously and 30 ml of aqueous NaOH was added. The mixture was filtered and the product was collected on filter paper. The powder was vacuum dried in an oven at 120 °C to give 4 (51.0 g).

MS Da/e = 215 (M + 1).

Calcd for _C8H7N2SOCl : C, _44.72 ; H, 3.29; N, 13.05; S, 14.93. Found: C, 44.63; H, 3.30; N, 12.96; S, 14.83.

(±)-2-(((2-Amino-4-chloro-5-methoxyphenyl)thio)methyl)-2-hexanoic acid (5)

To a suspension of 4 (20.0 g) in H ₂ O (200 mL) was added KOH (100 g). The slurry was heated at reflux for 7 h and allowed to cool to room temperature. To the dark solution was added 2 (33.2 g) in one portion. The reaction mixture was stirred for 18 h and the pH was adjusted to 4 with HCl. The mixture was transferred to a separatory funnel and extracted three times with ethyl acetate. The organic layer was dried and concentrated. The product was purified by column chromatography on silica gel eluting with 10% ethyl acetate/petroleum ether to give 5 (30.1 g).

./

MS Da/e = 368 (M + Na).

(±)-2,3-Dihydro-3-ethyl-3-butyl-5H-7-chloro-8-methoxy-1,5-benzothiazepin-4-one (6)

Toluenesulfonic acid (3.2 g) was added to a suspension of 5 (72.0 g) in tetradecane (900 ml). The mixture was heated at reflux for 15 min and 4 ml of water was collected in a Dean-Stark trap. The solution was cooled and transferred to a 1 L Erlenmeyer flask and allowed to stand for 18 h at 4 °C. The tetradecane was decanted and the solids obtained were recrystallized from methanol/water. The mother liquors and tetradecane solution were subjected to column chromatography on silica gel and the remaining product was eluted with 20% ethyl acetate/petroleum ether. The chromatographic material was combined with the recrystallized material to give 6 (52.6 g).

MS Da/e = 350 (M + Na).

Calcd for C16H22NSO2Cl: C, 58.61; H, 6.76; N, 4.27; S, 9.78. Found: C,

58.70; H, 6.82; N, 4.23; S, 9.82.

(3R)-2,3-Dihydro-3-ethyl-3-butyl~5H-7-chloro-8-methoxy-1,5-benzothiazepin-4-one (7)

Racemic 6 (50 g) was resolved on a CIRALPAK AD™ 10 x 50 cm column eluting with 100% methanol at 25 °C. The s-isomer eluted first, the r-isomer eluted second. After the concentration of the second peak decreased, 7 (23.28 g, 99% ee) was isolated.

¹ H NMR (CDCb) d 7.82 (s, 1H), 7.03 (s, 1H), 6.99 (s, 1H), 3.88 (s, 3H), 2.95 (s, 2H),

1.85 - 1.45 (m, 4H), 1.25 (m, 4H), 0.86 (m, 6H).

(3R)-2,3-Dihydro-3-ethyl-3-butyl-5-phenyl-7-chloro-8-methoxy-1,5-benzothiazepin-4-one (8)

To a solution of 7 (10.0 g) in iodobenzene (75 mL) was added copper (I) iodide (0.30 g) and potassium carbonate (4.23 g). The mixture was heated at reflux for 5.5 h and then allowed to cool to room temperature. The reaction mixture was transferred directly to a silica gel column. The iodobenzene was eluted with petroleum ether and the product was eluted with 15% ethyl acetate/petroleum ether to give 8 (10.9 g).

MS Da/e = 404 (M + 1), 426 (M + Na).

Calcd for C22H26NSO2Cl; C, 65.41; H, 6.49; N, 3.47; S, 7.94. Found: C, 65.15; H, 6.59; N, 3.34; S, 7.72.

• · · · · ♦ • · · · · ····· · · · · • ' ·· ·····

.......... ·· ·· ·· (3S)-2,3,4,5-Tetrahydro-3-ethyl-3-butyl-5-phenyl-7-chloro-8-methoxy-1,5benzothiazepine-1,1-dioxide (9)

To a 1 H solution of lithium aluminum hydride in ethyl ether (91.5 mL) was added dropwise at 0 °C a 7.2 M solution of sulfuric acid in THF (6.4 mL) and the mixture was stirred at 0 °C for 1 h. To the mixture was added 8 (10.9 g) in THF (75 mL) at 0 °C. The reaction mixture was allowed to warm to room temperature and stirred for 3.5 h at room temperature and then cooled back to 0 °C and a 30% (v/v) solution of HO in THF was added dropwise. Then 1 N NaOH solution (15 mL) was added. The reaction mixture was filtered through a fritted funnel to remove aluminum oxides. The filtrate was transferred to a separatory funnel and partitioned between water and ethyl ether. The aqueous layer was extracted three times with ether. The organic phase was dried (Na2SO4) and concentrated. The resulting oil was transferred to THF (175 mL). To the KTHF solution was added tert-butanol (60 mL), N-methylmorpholine-N-oxide (10.7 g) and osmium tetroxide (2.5 wt% in tert-butanol, 7.6 mL). The reaction mixture was stirred for 18 h at room temperature. The reaction mixture was transferred to a separatory funnel and partitioned between brine and ethyl acetate. The aqueous layer was extracted three times with ethyl acetate. The organic layer was dried, concentrated and the residue was applied to a silica gel column. The product was eluted with 10% ethyl acetate/petroleum ether to give 9 (10.92 g). mp 147.5 °C.

' ’ MS Da/e = 422 (M + 1), 444 (M + Na).

Calcd for C22H28NSO3Cl: C, 62.62; H, 6.69; N, 3.32; S, 7.60. Found: C, 62.53; H, 6.62; N, 3.32; S, 7.53.

(3S)-2,3,4,5-Tetrahydro-3-ethyl-3-butyl-5-phenyl-7-chloro-8-hydroxy-1,5-benzothiazepine-1,1-dioxide (Example 1)

To a solution of 9 (10.92 g) in methylene chloride (150 ml) at 0 °C was added a 1 M solution of boron tribromide in methylene chloride (36.3 ml). The reaction mixture was slowly warmed to room temperature and stirred for 18 hours before being cooled back to 0 °C and water (100 ml) was added dropwise. The mixture was transferred to a separatory funnel and extracted three times with methylene chloride. The organic extracts were dried, concentrated and the residue was applied to a silica gel column. The product was eluted with 30% ethyl acetate/petroleum ether to give the compound of Example 1 (10.12 g).

Melting point 179.6 to 180.2 °C.

MS Da/e = 406 (M - 1, negative ion mode).

···· · · ·«

Calcd for C12H26NSO3Cl: C, 61.83; H, 6.42; N, 3.43; S, 7.86. Found: C,

61.76; H, 6.47; N, 3.37; S, 7.76.

Biological test

Inhibition of bile acid reabsorption in vivo

Male Sprague-Dawley rats (CD; Charles River); weighing 220.0-260.0 g, were housed in individual cages and fed a standard diet. The rats were administered test compounds (1 ml/100 g body weight) as a suspension in 0.5% methylcellulose by oral gavage at 9.0 a.m. and 3.30 p.m. for 2 days. The control group of rats was administered 0.5% methylcellulose alone. On the second day, 2.0 hours after the morning dose, the rats were orally administered a trace amount (1.30 nmol) of ^23.25-75 Se tauromonocholate ( ⁷⁵ SeHCAT) in 1 ml saline. A synthetic bile acid analogue, ⁷⁵ SeHCAT, which emits gamma rays and is absorbed by a bile acid absorption system similar to that of taurochloric acid, is used clinically to monitor bile acid absorption in the intestinal tract. Feces were collected for 24 hours after administration of ⁷⁵ SeHCAT. The content of ⁷⁵ SeHCAT in the feces was determined using a gamma-particle counter (Packard Auto-Gamma 5000 series). The percentage inhibition of bile acid reabsorption is calculated according to the following formula:

% inhibition = total ⁷⁵ SeHCAT content - excluded ⁷⁵ SeHCAT content in the sample minus---------------------------------------------------------------------------------------------------- x 100 total ⁷⁵ SeHCAT content - excluded ⁷⁵ SeHCAT content in the control

The percentage inhibition of bile acid reabsorption in rats using ⁷⁵ SeHCAT was used to determine the ED30 (dose required for 30% inhibition of bile acid absorption).

• » · · · · · · · · · · • · ···· · · · « · · · ······ · · · · • » ·· ·····

2,3,4,5-Tetrahydro-3-ethyl-3-butyl-5-phenyl-7-chloro-8-hydroxy-1,5-benzothiazepine-1,1-dioxide (Example 1 of the present invention) and the corresponding bromo compound, 2,3,4,5tetrahydro-3-ethyl-3-butyl-5-phenyl-7-bromo-8-hydroxy-1,5-benzothiazepine-1,1-dioxide (Example A, as described in PCT/GB/02700) were tested directly in two series of experiments with 6 rats in each set (n = 12 in total).

^ED30 (mg/kg)

Example 1 0 048

Example A 0.17 (II) Reduction of percentage cholesterol in rats

Hypercholesterolemia was induced in Spraque-Dawley rats (CD, Charles River, weighing 200 to 300 g) by feeding a diet enriched with cholesterol and cholic acid. The diet was prepared from Wayne Laboratory Lab Blocks by grinding into the diet and mechanically mixing with powdered cholesterol and cholic acid to a final concentration (by weight) of 1% and 0.5%, respectively. Before feeding, blood was collected by cardiac puncture under halothane anesthesia to determine baseline lipid levels. The serum obtained was used to determine total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), and dextran-coagulated lipoprotein cholesterol (VLDL+LDL). Rats were divided into groups so that each group had similar baseline serum lipid levels. Five days after the initial serial lipid sampling, rats were fed a diet enriched with cholesterol and cholic acid ad libitum and administration of the test compound began. The compound was administered by gavage as a suspension in 0.5% methylcellulose (1 ml/100 g body weight) twice daily at 9:00 AM and 3:00 PM for 3 days and at 9:00 AM on the fourth day. Control animals received 0.5% cellulose alone. Blood was collected from rats 4 hours after the last dose for determination of serum lipid content. All collections were made after a 4-hour fast. Serum TC concentrations were determined enzymatically using reagents obtained from Seragen Diagnostics (2). Serum HDL-C was determined after selective precipitation of VLDL and LDL with dextran sulfate and magnesium sulfate, reagents obtained from Seragen (3). HDL-C was determined in the supernatant. VLDL + LDL cholesterol was determined as the difference between total and HDL-C. The following results were obtained for Example 1 and Example A defined above.

0.3 Dose (mg/kg) 0.1 0.03 Example 1 81% 56% 48% Example A 53% 17% 14%

The results clearly demonstrate the unexpected cholesterol-lowering properties of the compounds of the invention.

Examples of pharmaceuticals

In the following examples, the active compound may be any compound of formula I and/or a pharmaceutically acceptable salt, solvate or physiologically functional derivative thereof.

(I) Composition of tablets

The following compositions A and B can be prepared by wet granulation of components (a) to (c) and (a) to (d) with a povidone solution, followed by addition of magnesium stearate and compression.

Means A

(a) Active ingredient mq/tablet 250 mg/tablet 250 (b) Lactose B.P. 210 26 (c) Sodium starch glycolate 20 12 (d) Povidone B.P. 15 9 (e) Magnesium stearate 5 3 500 300

• · · « ·

Means B

mq/tablet mq/tablet (a) Active ingredient 250 250 (b) Lactose 150 150 - (c) Avicel PH 101 60 26 (d) Sodium starch glycolate 20 12 (e) Povidone B.P. 15 9 (f) Magnesium stearate 5 3 500 300 Method C mq/tablet Active ingredient 100 Lactose 200 Starch 50 Povidone 5 Magnesium stearate 4

359

Active ingredient Magnesium stearate Pregelatinized starch NF15

The following compositions D and E can be prepared by direct compression of the mixed ingredients. The lactose used in composition E is of the type intended for direct compression.

Agent D mg/tablet

250 4

146,400

Method E mo/tablet Active ingredient 250 Magnesium stearate 5 Lactose 145 Avicel 100 500 Formula F (controlled release formulation) mg/tablet (a) Active ingredient 500 (b) Hydroxypropylmethylcellulose (Methocel K4M Premium) 112 (c) Lactose B.P. 53 (d) Povidone B.P.C. 28 (e) Magnesium stearate 7 700

The compositions must be prepared by wet granulation of components (a) to (c) with a povidone solution, followed by addition of magnesium stearate and compression.

Preparation G (enteric-coated tablet)

Enteric coated tablets of Formulation C can be prepared by coating the tablets with a suitable enteric polymer such as cellulose acetate phthalate, polyvinyl acetate phthalate, hydroxypropyl methylcellulose phthalate or anionic polymers of methacrylic acid and methacrylic acid methyl esters (Eudragit L) in an amount of 25 mg/tablet. In addition to Eudragit L, these polymers should also include 10% (by weight based on the amount of polymer used) of a plasticizer to prevent cracking of the coating during administration or storage. Suitable plasticizers include diethyl phthalate, tributyl citrate and triacetin.

Formulation H (enteric-coated controlled-release tablet) • · · · · · · · · · · ·

Enteric coated tablets of Formulation F can be prepared by coating the tablets with a suitable enteric polymer such as cellulose acetate phthalate, polyvinyl acetate phthalate, hydroxypropyl methylcellulose phthalate or anionic polymers of methacrylic acid and methacrylic acid methyl ester (Eudragit L) in an amount of 50 mg/tablet. In addition to Eudragit L, these polymers should also include 10% (by weight based on the amount of polymer used) of a plasticizer to prevent cracking of the coating during administration or storage. Suitable plasticizers include diethyl phthalate, tributyl citrate and triacetin.

(II) Capsule preparations

Means A

Capsules can be prepared by mixing the ingredients of Formulation D above and filling two-piece hard gelatin capsules with the resulting mixture. Formulation B (infra) can be prepared in a similar manner.

Formulation B mg/capsule (a) Active ingredient 250 (b) Lactose B.P. 143 (c) Sodium stearate 25 (d) Magnesium stearate 2

420

Formulation C mq/capsule (a) Active ingredient 250 (b) Macrogol 4000 BP 350

600

Capsules can be prepared by melting Macrogol 4000 BP, dispersing the active ingredient in the melt and filling two-piece hard gelatin capsules with the resulting mixture.

• · ·

Agent D mg/capsule

Active ingredient 250

Lecithin 100

Peanut oil 100

450

Capsules can be prepared by dispersing the active ingredient in lecithin and peanut oil and filling soft, elastic gelatin capsules with this dispersion.

Formulation E (controlled release capsules) mg/capsule (a) Active ingredient 250 (b) Microcrystalline cellulose 125 (c) Lactose BP 125 (d) Ethylcellulose 13

513

The controlled release capsule formulation can be prepared by extruding the mixed components (a) to (c) using an extruder, then spheronizing and drying the extrudate. The dried beads are coated with a release controlling coating (d) and filled into two-piece hard gelatin capsules.

Formulation F (Enteric-coated capsules) mg/capsule (a) Active ingredient 250 (b) Microcrystalline cellulose 125 (c) Lactose BP 125 (d) Cellulose acetate phthalate 50 (e) Diethyl phthalate 5

555

The enteric capsule formulation can be prepared by extruding the mixed components (a) to (c) using an extruder, then rounding and drying the extrudate. The dried beads are coated with an enteric coating (d) containing a plasticizer (e) and filled into two-piece hard gelatin capsules.

«··· · · · ·

Formulation G (enteric-coated controlled-release capsules)

Enteric-coated capsules of Formulation E can be prepared by coating controlled-release beads with enteric polymers such as cellulose acetate phthalate, polyvinyl acetate phthalate, hydroxypropyl methylcellulose phthalate or anionic polymers of methacrylic acid and methacrylic acid methyl ester (Eudragit L) at 50 mg/capsule. In addition to Eudragit L, these polymers should also include 10% (w/w based on the amount of polymer used) of a plasticizer to prevent cracking of the coating during administration or storage. Suitable plasticizers include diethyl phthalate, tributyl citrate and triacetin.

(III) Intravenous injection device

Active ingredient 0.200 g

Sterile, pyrogen-free phosphate buffer (pH 9.0) up to 10 ml

The active ingredient is dissolved in most of the phosphate buffer at 35 to 40 °C, made up to the required volume and filtered through a sterile microporous filter into sterile 10 ml glass vials (type 1), which are closed with sterile closures and caps.

(IV) Intramuscular injection device

Active ingredient 0.20 g

Benzyl alcohol 0.10 g

Glycofurol 75 1.45 g

Water for injections up to 3.00 ml

The active ingredient is dissolved in glycofurol. Benzyl alcohol is then added and dissolved and water is added to make up to 3 ml. The mixture is filtered through a sterile microporous filter and sealed in sterile 3 ml glass vials (type 1).

• · · · · * · ·· · ····· ·· ···· · · · · (V) Syrup

Active ingredient 0.25 g

Sorbitol solution 1.50 g

Glycerol 1.00 g

Sodium benzoate 0.005 g

Flavorings 0.0125 ml

Purified water q.s. up to 5.0 ml

Sodium benzoate is dissolved in a portion of purified water and sorbitol solution is added. The active ingredient is added and dissolved. The resulting solution is mixed with glycerol and made up to the desired volume with purified water.

(VI) Suppository mq/suppository

Active ingredient 250

Hard Fat, BP (Witepsol H15-1770 Dynamit NoBel) 1770 2020

One fifth of the Witepso H15 fat is melted in a pan with a steam jacket at a maximum of 45°C. The active ingredient is sieved through a 200 µm sieve and added to the molten base while stirring with a Silverson mixer with a cutting head until a smooth dispersion is obtained. While maintaining the temperature at 45°C, the remaining fat is added to the suspension with constant stirring until a homogeneous mixture is obtained. The suspension is then passed through a 250 µm stainless steel sieve and allowed to cool to 40°C with constant stirring. At a temperature of 38 to 40°C, portions of the mixture weighing 2.02 g are filled into suitable plastic moulds and the suppositories are allowed to cool to room temperature.

(VII) Pessary

Active ingredient (63 Im) mg/pesary 250 Dextrose Anhydrous 380 Potato starch 363 Magnesium stearate 7 1000

• · · · ····

The listed ingredients are mixed directly and the pessaries are prepared by pressing the resulting mixture.

vii) Transdermal device

Active ingredient 200 mg

Alcohol USP 0.1 ml

Hydroxyethylcellulose

The active ingredient and alcohol USP are gelled with hydroxyethylcellulose and packaged in a transdermal applicator with a surface area of 10 cm ² .

?V AooO - 2SSI

Claims (11)

PATENT REQUIREMENTS where R ¹ is H or methyl; or a salt, solvate or physiologically functional derivative thereof. 2. A compound of formula I selected from the group consisting of: (±) -2,3,4,5-Tetrahydro-3-ethyl-3-butyl-5-phenyl-7-chloro-8-hydroxy-1,5-benzothiazepine-1,1-dioxide; (3S) -2,3,4,5-Tetrahydro-3-ethyl-3-butyl-5-phenyl-7-chloro-8-hydroxy-1,5-benzothiazepine-1,1-dioxide; and (±) -2,3,4,5-Tetrahydro-3-ethyl-3-butyl-5-phenyl-7-chloro-8-methoxy-1,5-benzothiazepine-1,1-dioxide; or a salt, solvate or physiologically functional derivative thereof. 3. (±) -2,3,4,5-Tetrahydro-3-ethyl-3-butyl-5-phenyl-7-chloro-8-hydroxy-1,5-benzothiazepine-1,1-dioxide or a salt thereof, solvate or a physiologically functional derivative. 4. (3S) -2,3,4,5-Tetrahydro-3-ethyl-3-butyl-5-phenyl-7-chloro-8-hydroxy-1,5-benzothiazepine-1,1-dioxide or a salt thereof, solvate or a physiologically functional derivative. 5. A method of treating hyperlipidemic conditions in a mammal comprising administering to the mammal an amount effective for hypelipidemic treatment of a compound of formula I as claimed in any one of claims 1 to 4, or a pharmaceutically acceptable salt, solvate or physiologically functional derivative thereof. • · · · · · · · · · · · · · · The method of claim 5, wherein the hyperlipidemic condition is atherosclerosis. A compound of formula I as claimed in any one of claims 1 to 4, or a pharmaceutically acceptable salt, solvate or physiologically functional derivative thereof for use in medicine. A compound of formula I as claimed in any one of claims 1 to 4, or a pharmaceutically acceptable salt, solvate or physiologically functional derivative thereof, for use in the prophylaxis or treatment of a hyperlipidemic condition. A compound of formula I as claimed in any one of claims 1 to 4 or a pharmaceutically acceptable salt, solvate or physiologically functional derivative thereof for use in the manufacture of a medicament for the treatment of a hyperlipidemic condition. 10. A pharmaceutical composition comprising a compound of formula I as claimed in any one of claims 1 to 4, or a pharmaceutically acceptable salt, solvate or physiologically functional derivative thereof, at least one carrier and optionally one or more physiologically active agents. 11. A process for the preparation of a compound of formula (I) or a salt, solvate or physiologically functional derivative thereof comprising: (A) when R ¹ is H, dealkylation of a compound of Formula II wherein R ^1a is an alkyl moiety; or (II) (B) when R ¹ is methyl, oxidation of a compound of formula III (III)