RU2282628C2 - Derivatives of diphenylazethidinone, pharmaceutical composition based on thereof and method for its preparing - Google Patents

Abstract

FIELD: organic chemistry, pharmacy.

SUBSTANCE: invention relates to novel derivatives of diphenylazethidinone of the formula (I):

wherein R1 means hydrogen atom; R₂ means -(CH₂)_0-1-NH-C(O)-(C₃-C₁₂)-alkylene-C(O)NH-L; R₃ and R₆ mean hydrogen atom; R₄ and R₅ mean atoms F, Cl, Br, J; L means compound of the formula:

wherein R_x, R_y and R_z mean hydrogen atom (H), and to their pharmaceutically acceptable salts also. Compounds of the formula (I) elicit cholesterol-declining activity and can be used in medicine.

EFFECT: improved preparing method, valuable medicinal properties of compounds and composition.

4 cl, 9 ex

Description

The invention relates to substituted diphenylazetidinones, their physiologically compatible salts, as well as physiologically functional derivatives.

Diphenylazetidinones (for example, ezetimibe), as well as their use for the treatment of hyperlipidemia, as well as arteriosclerosis and hypercholesterolemia, have already been described [see Drugs of the Future 2000, 25 (7): 679-685].

The objective of the invention is to obtain new compounds with therapeutically suitable lipid-lowering effect. The objective of the invention mainly consisted in the search for new compounds, which, compared with compounds known from the prior art, differ by very little resorption. By very slight resorption is meant intestinal resorption of less than 10%, mainly less than or equal to 5%. In particular, new compounds should have lower resorption than ezetimibe.

With low resorption, pharmaceutically active substances, as a rule, exhibit significantly fewer side effects.

The invention relates to compounds of formula I

in which mean

-N((C₁-C₆)алкил)- или -NH-,R1, R2, R3, R4, R5, R6 are independently (C ₀ -C ₃₀ ) alkylene-L, and one or more C-atoms of the alkylene residue may be substituted with -O-, - (C = O) - , -CH = CH-,

-N ((C ₁ -C ₆ ) alkyl) - or -NH-,

H, F, Cl, Br, I, CF ₃ , NO ₂ , CN, COOH, COO (C ₁ -C ₆ ) alkyl, CONH ₂ , CONH (C ₁ -C ₆ ) alkyl, CON [(C ₁ -C ₆ ) alkyl] ₂ , (C ₁ -C ₆ ) alkyl, (C ₂ -C ₆ ) alkenyl, (C ₂ -C ₆ ) alkynyl, O- (C ₁ -C ₆ ) alkyl, with one in the alkyl residue, some or all of the hydrogen atoms may be substituted by fluorine;

SO ₂ -NH ₂ , SO ₂ NH (C ₁ -C ₆ ) alkyl, SO ₂ N [(C ₁ -C ₆ ) alkyl] ₂ , S- (C ₁ -C ₆ ) alkyl, S- (CH ₂ ) _n- phenyl, SO- (C ₁ -C ₆ ) alkyl, SO- (CH ₂ ) _n- phenyl, SO ₂ - (C ₁ -C ₆ ) alkyl, SO ₂ - (CH ₂ ) _n- phenyl, and n = 0-6 and the phenyl residue can be substituted once or twice by F, Cl, Br, OH, CF ₃ , NO ₂ , CN, OCF ₃ , O (C ₁ -C ₆ ) alkyl, (C ₁ -C ₆ ) alkyl , NH ₂ group,

NH ₂ , NH- (C ₁ -C ₆ ) alkyl, N ((C ₁ -C ₆ ) alkyl) ₂ , NH (C ₁ -C ₇ ) acyl, phenyl, O- (CH ₂ ) _n- phenyl, moreover the phenyl ring may be substituted once, twice or three times with F, Cl, Br, I, OH, CF ₃ , NO ₂ , CN, OCF ₃ , O- (C ₁ -C ₆ ) alkyl, (C ₁ -C ₆ ) alkyl, NH ₂ , NH (C ₁ -C ₆ ) alkyl, N ((C ₁ -C ₆ ) alkyl) ₂ , SO ₂ -CH ₃ , COOH, COO- (C ₁ -C ₆ ) alkyl, CONH ₂ ;

Rx, Ry, Rz are independently H, F, Cl, Br, I, CF ₃ , NO ₂ , CN, COOH, COO (C ₁ -C ₆ ) alkyl, CONH ₂ , CONH (C ₁ -C ₆ ) alkyl, CON [(C ₁ -C ₆ ) alkyl] ₂ , (C ₁ -C ₆ ) alkyl, (C ₂ -C ₆ ) alkenyl, (C ₂ -C ₆ ) alkynyl, O- (C ₁ -C ₆ ) alkyl, and in the alkyl residue one, several or all hydrogen atoms can be substituted by fluorine;

SO ₂ -NH ₂ , SO ₂ NH (C ₁ -C ₆ ) alkyl, SO ₂ N [(C ₁ -C ₆ ) alkyl] ₂ , S- (C ₁ -C ₆ ) alkyl, S- (CH ₂ ) _n- phenyl, SO- (C ₁ -C ₆ ) alkyl, SO- (CH ₂ ) _n- phenyl, SO ₂ - (C ₁ -C ₆ ) alkyl, SO ₂ - (CH ₂ ) _n- phenyl, and n = 0-6 and the phenyl residue can be substituted once or twice by F, Cl, Br, OH, CF ₃ , NO ₂ , CN, OCF ₃ , O- (C ₁ -C ₆ ) alkyl, (C ₁ -C ₆ ) alkyl, NH ₂ ;

NH ₂ , NH (C ₁ -C ₆ ) alkyl, N ((C ₁ -C ₆ ) alkyl) ₂ , NH (C ₁ -C ₇ ) acyl, phenyl, O- (CH ₂ ) _n- phenyl, with n = 0-6, moreover, the phenyl ring one, two or three times can be substituted by F, Cl, Br, I, OH, CF ₃ , NO ₂ , CN, OCF ₃ , O- (C ₁ -C ₆ ) alkyl, (C ₁ -C ₆ ) alkyl, NH ₂ , NH (C ₁ -C ₆ ) alkyl, N ((C ₁ -C ₆ ) alkyl) ₂ , SO ₂ -CH ₃ , COOH, COO- (C ₁ -C ₆ ) alkyl, CONH ₂ ;

-N((C₁-C₆)алкил)- или -NH-,moreover, at least one of the radicals R1-R6 must always have the value (C ₀ -C ₃₀ ) alkylene-L, moreover, one or more C-atoms of the alkylene residue can be substituted by groups -O-, - (C = O) -, -CH = CH-,

-N ((C ₁ -C ₆ ) alkyl) - or -NH-,

and their pharmaceutically acceptable salts.

Preferred are compounds of formula I in which at least one of the radicals R1-R6 has the meaning (C ₀ -C ₃₀ ) alkylene-L, wherein one or more C-atoms of the alkylene residue can be substituted with the groups —O—, - (C = O) - or -NH-,

Particularly preferred are compounds of formula I in which one of the radicals R1 or R3 has the meaning (C ₀ -C ₃₀ ) alkylene-L, wherein one or more C-atoms of the alkylene radical can be substituted with the groups —O—, - (C = O) - or -NH-.

And completely preferred are compounds of formula I in which one of the radicals R1 or R3 has the meanings - (CH ₂ ) _0-1 -NH- (C = O) _0-1 - (C ₃ -C ₂₅ ) alkylene- (C = O ) _0-1- NH-L, wherein one or more C-atoms of the alkylene moiety may be substituted with an O-atom.

The binding of one of the residues R1-R6 to the L-residue occurs mainly in the meta-position of the ring C of the L-group.

Pharmaceutically acceptable salts, due to their high solubility in water compared to the starting or basic compounds, are particularly suitable for medical use. These salts must contain pharmaceutically compatible anions and cations. Pharmaceutically acceptable complex acid salts of the compounds of the invention are salts of inorganic acids such as hydrochloric acid, hydrobromic, phosphoric, metaphosphoric, nitric, sulfonic and sulfuric acids, as well as organic acids such as, for example, acetic, benzenesulfonic, benzoic , citric, ethanesulfonic, fumaric, gluconic, glycolic, isothionic, lactic, lactobionic, maleic, malic, methanesulfonic, succinic, p-toluenesulfonic, tartaric and trifluoroacetic acids. For medical purposes, a method in which chlorine salt is used is particularly preferred. Suitable pharmaceutically acceptable base salts are ammonium salts, alkali metal salts (such as sodium and potassium salts) and alkaline earth metal salts (such as magnesium and calcium salts).

Salts with a pharmaceutically unacceptable anion also belong to this invention as useful intermediates for the preparation or purification of pharmaceutically acceptable salts and / or for use for non-therapeutic purposes, for example, for in vitro use.

As used herein, “physiologically functional derivatives” means each physiologically acceptable derivative of a compound of the invention, for example, an ester which, when administered to a mammal, such as a human, (directly or indirectly) forms such a compound or active metabolite.

One of the following objects of this invention are prodrugs (Prodrugs) of the compounds according to the invention. Such prodrugs can be metabolized in vivo to the compounds of the invention. Such prodrugs alone may or may not be effective.

The compounds of the invention can also exist in various polymorphic forms, for example, in the form of amorphous and crystalline polymorphic forms. All polymorphic forms of the compounds according to the invention relate to this invention and are objects of the invention.

All subsequent references to “compound (s) according to formula (I)” extend to “compound (s) according to formula (I)” as described previously, as well as their salts, solvates and physiologically functional derivatives, as described above.

The compounds of formula I and their pharmaceutically acceptable salts and physiologically functional derivatives are ideal drugs for the treatment of metabolic disorders of lipid substances, especially hyperlipidemia. The compounds of formula I are also suitable for regulating serum cholesterol, as well as for the prevention and treatment of arteriosclerotic phenomena.

The compound (s) of formula (I) may be taken in conjunction with other active substances.

The amount of the compound of formula (I) that is necessary to achieve the desired biological effect depends on a number of factors, for example, on the particular compound selected, purpose of use, route of administration and clinical condition of the patient. Basically, the daily dose is in the range from 0.1 to 100 mg (usually 0.1-50 mg) per day per kilogram of body weight, for example, 0.1-10 mg / kg / day. Tablets and capsules may contain, for example, from 0.01 to 100 mg, usually 0.02-50 mg. In the case of pharmaceutically acceptable salts, the named weight amounts refer to the weight of the benzotiepine ion included in the salt. For the prevention and treatment of the above conditions, the compounds of formula (I) can be used in pure form, however, it is preferable to use them together with acceptable carriers in the form of a pharmaceutical composition. The carrier should naturally be acceptable in the sense that it must be compatible with other components of the composition and not adversely affect the health of the patient. The carrier may be a solid or liquid, or in the form of both, and is preferably used together with the compound to produce a single unit dose, for example, in the form of a tablet, which may contain from 0.05 to 95% of the active substance. Other pharmaceutically active substances may be present, including other compounds of formula (I). The pharmaceutical compositions according to the invention can be manufactured by known pharmaceutical methods, which mainly consist in the fact that the components are mixed with pharmacologically acceptable carriers and / or excipients.

The pharmaceutical compositions according to the invention are suitable for oral and oral (e.g., sublingual) administration, although the suitable route of administration depends in each individual case on the type and severity of the disease being treated and on the type of the compound of formula (I) used. The subject of the invention is also dragee pharmaceutical preparations and dragee pharmaceutical preparations of prolonged action. Acidic and gastric juice resistant formulations are preferred. Suitable gastric juice resistant coatings are cellulose acetate phthalate, polyvinyl acetate phthalate, hydroxypropyl methyl cellulose phthalate and anionic polymers of methacrylic acid and methyl methacrylate.

Suitable pharmaceutical compounds for oral administration may be in unit dosage form, for example, in the form of capsules, cachet capsules, sucking tablets or tablets that contain specific amounts of a compound of formula (I); in the form of powder or granules; in the form of a solution or suspension in an aqueous or non-aqueous liquid; or as an oil-in-water emulsion or water-in-oil emulsion. Such compositions, as already mentioned, can be prepared by any suitable pharmaceutical method, which includes the stages in which the active substance and the carrier (which may consist of one or more components) are brought into contact. In general, compositions are prepared by uniformly and homogeneously mixing the active substance with a liquid and / or finely divided solid carrier, after which the product is formed, if necessary. Thus, for example, a tablet can be made, during which the powder or granulate of the compound is compressed or molded, if necessary with one or more additional components. Compressed tablets may be made by tabletting the compound in a free-flowing form, such as, for example, in the form of powder or granules, optionally mixed with a binder, a glidant, inert diluents and / or one or more surfactant / dispersant in special apparatuses. Molded tablets can be made in special apparatuses by molding a powdered compound moistened with an inert liquid diluent.

Pharmaceutical formulations suitable for oral (sublingual) administration include sucking tablets containing a compound of formula (I) with flavoring agents, usually sucrose and gum arabic or tragacanth, and lozenges that contain an inert base such as gelatin and glycerin or sucrose and gum arabic .

Suitable other active ingredients for combination preparations are: all antidiabetic agents listed in the 2001 red list, chapter 12. They can be combined with the compounds of formula I according to the invention, especially for synergistic improvement of the effect. Reception of combined preparations can be carried out both by separate administration of active substances to the patient, and in the form of combined preparations in which several active substances are in the same pharmaceutical preparation.

Antidiabetic agents include insulin and insulin derivatives, such as, for example, Lantus® or HMR 1964, GLP-1 derivatives, as, for example, published in WO 98/08871 from Novo Nordisk A / S, as well as oral hypoglycemic actives.

Oral hypoglycemic actives include predominantly sulfonylurea, biguadine, meglitinide, oxadiazolidinone, thiazolidinedione, glucosidase inhibitors, glucagon antagonists, GLP-1 agonists, potassium channel openers, such as, for example, those published in patent applications WO 97/26502 Nordisk A / S, insulin-sensitizing substances, liver enzyme inhibitors that are involved in stimulating gluconeogenesis and / or glucogenolysis, glucose uptake modulators, lipid-modifying compounds such as antihyperlipidemic skie active ingredients and antilipidemic active ingredients, compounds which reduce the absorption of food, PPAR- and PXR-agonists and active agents affecting on the ATP-dependent potassium channel of the beta cells.

In one embodiment of the invention, the compounds of formula I are administered in combination with HMGCoA reductase inhibitors such as simvastatin, fluvastatin, pravastatin, lovastatin, atorvastatin, cerivastatin, rosuvastatin.

In one embodiment of the invention, the compounds of formula I are administered in combination with a cholesterol resorption inhibitor, such as, for example, ezetimibe, tikvesides, pamakvesides.

In one embodiment of the invention, the compounds of formula I are used in combination with a PPAR gamma agonist, such as, for example, rosiglitazone, pioglitazone, JTT-501, Gl 262570.

In one embodiment of the invention, the compounds of formula I are used in combination with a PPAR alpha agonist, such as, for example, GW 9578, GW 7647.

In one embodiment of the invention, the compounds of formula I are used in combination with a PPAR alpha / gamma agonist, such as, for example, GW 1536, AVE 8042, AVE 8134, AVE 0847.

In one embodiment of the invention, the compounds of formula I are used in combination with fibrate, such as fenofibrate, clofibrate, bezafibrate.

In one embodiment of the invention, the compounds of formula I are used in combination with an MTP inhibitor, such as, for example, Bay 13-9952, BMS-201038, R-103757.

In one embodiment of the invention, the compounds of formula I are used in combination with a bile acid resorption inhibitor, such as, for example, HMR 1453.

In one embodiment of the invention, the compounds of formula I are used in combination with a CETP inhibitor, such as, for example, Bay 194789.

In one embodiment of the invention, the compounds of formula I are used in combination with a polymeric bile acid absorber, such as, for example, cholestyramine, cholavol.

In one embodiment of the invention, the compounds of formula I are used in combination with an LDL receptor inducer, such as, for example, HMR1171, HMR1586.

In one embodiment of the invention, the compounds of formula I are used in combination with an ACAT inhibitor, such as, for example, avazimib.

In one embodiment of the invention, the compounds of formula I are used in combination with an antioxidant, such as, for example, OPC-14117.

In one embodiment of the invention, the compounds of formula I are administered in combination with a lipoprotein lipase inhibitor, such as, for example, NO-1886.

In one embodiment of the invention, the compounds of formula I are administered in combination with an ATP-citrate lyase inhibitor, such as, for example, SB-204990.

In one embodiment of the invention, the compounds of formula I are administered in combination with a squalene synthetase inhibitor, such as, for example, BMS-188494.

In one embodiment of the invention, the compounds of formula I are used in combination with a lipoprotein antagonist, such as, for example, Cl-1027 or nicotinic acid.

In one embodiment of the invention, the compounds of formula I are used in combination with a lipase inhibitor, such as orlistat.

In one embodiment of the invention, the compounds of formula I are used in combination with insulin.

In one embodiment of the invention, the compounds of formula I are used in combination with a sulfonylurea, such as tolbutamide, glibenclamide, glipizide or glycazide.

In one embodiment of the invention, the compounds of formula I are used in combination with a biguanide, such as metformin.

In one embodiment of the invention, the compounds of formula I are used in combination with meglitinide, such as, for example, repaglinide.

Research Foundation, особенно 5-[[4-[(3,4-дигидро-3-метил-4-оксо-2-хиназолинилметокси]фенил]метил]-2,4-тиазолидиндион.In one embodiment of the invention, the compounds of formula I are used in combination with thiazolidinedione, such as troglitazone, ciglitazone, pioglitazone, rosiglitazone or with the compounds disclosed in WO 97/41097 Dr.

Research Foundation, especially 5 - [[4 - [(3,4-dihydro-3-methyl-4-oxo-2-quinazolinylmethoxy] phenyl] methyl] -2,4-thiazolidinedione.

In one embodiment of the invention, the compounds of formula I are used in combination with an α-glucosidase inhibitor, such as, for example, miglitol or acarbose.

In one embodiment of the invention, the compounds of formula I are used in combination with an active substance that acts on the ATP-dependent potassium channel of beta cells, such as tolbutamide, glibenclamide, glipizide, gliazide or repaglinide.

In one embodiment of the invention, the compounds of formula I are administered in combination with more than one of the above compounds, such as, for example, in combination with sulfonylurea and metformin, sulfonylurea and acarbose, repaglinide and metformin, insulin and sulfonylurea, insulin and metformin, insulin and troglitazone, insulin and lovastatin, etc.

In other embodiments, the compounds of formula I are administered in combination with CART agonists, NPY agonists, MC4 agonists, orexin agonists, H3 agonists, TNF agonists, CRF agonists, CRF BP antagonists, urocortin agonists, β3 agonists, melanocyte-stimulating hormone (MSH) agonists, CCK agonists, serotonin recovery inhibitors, mixed serotonin and noradrenergic compounds, 5HT agonists, bombesin agonists, galanin antagonists, growth hormones, growth hormone releasing compounds, H hormone releasing compounds protein-forming 2- or 3-modulators, leptin agonists, DA agonists (bromocreptin, doprexin), lipase / amylase inhibitors, PPAR modulators, RXR modulators or TR-β agonists.

In one embodiment of the invention, the additional active substance is leptin.

In one embodiment of the invention, the additional active substance is dexamphetamine or amphetamine.

In one embodiment of the invention, the additional active substance is fenfluramine or dexenfluramine.

In another embodiment of the invention, the additional active substance is sibutramine.

In one embodiment of the invention, the additional active substance is orlistat.

In one embodiment of the invention, the additional active substance is mazindol or phentermine.

In one embodiment of the invention, the compounds of formula I are used in combination with ballast materials, mainly insoluble ballast substances, such as, for example, Caromax®. The combination with Caromax® can be in the same dosage form or administered by separate administration of the compounds of formula I and Caromax®. Caromax® can also be administered in the form of food products, such as, for example, in baked goods or cereal flakes. The combination of compounds of Formula I with Caromax®, along with an improved action, is particularly distinguished by a decrease in LDL cholesterol compared to individual active substances, as well as its improved tolerance.

It should be borne in mind that each suitable combination of compounds according to the invention with one or more of the above compounds and optionally one or more other pharmacologically active substances are the subject of protection of this invention.

The invention further relates to stereoisomeric mixtures of formula I, as well as pure stereoisomers of formula I, as well as diastereomeric mixtures of formula I and pure diastereomers. The separation of mixtures occurs by chromatographic method.

Racemic as well as enantiomerically pure compounds of formula I of the following structure are preferred:

Further preferred are compounds of formula I in which the L moieties are as follows:

The invention further relates to a process for preparing compounds of general formula I, characterized in that compounds of formula I are prepared according to the following reaction scheme

-N((C₁-C₆)алкил)- или -NH.R4 ^" means (C ₀ -C ₃₀ ) alkylene, wherein one or more C-atoms of the alkylene residue may be substituted with -O-, - (C = O) -, -CH = CH-,

-N ((C ₁ -C ₆ ) alkyl) - or -NH.

Alternatively, binding to the L group occurs via ring A or ring C.

The following examples illustrate the invention in more detail, but do not limit it to the products described in the examples and the forms of execution.

Example IV

4- {1- (4-Fluorophenyl) -3- [3- (4-fluorophenyl) -3-hydroxypropyl] -4-oxoazetidin-2-yl} benzylamide {2- [3-hydroxy-3-phenyl-2- pyridin-2-yl-1- (pyridin-2-ylamino) propyl] phenyl} hexane acid amide ( 14 )

a) 5- {2- [3-Hydroxy-3-phenyl-2-pyridin-2-yl-1- (pyridin-2-ylamino) propyl] phenylcarbamoyl} pentanoic acid ( 13 )

This compound is prepared from 1.5 g of 3- (2-aminophenyl) -1-phenyl-2-pyridin-2-yl-3- (pyridin-2-ylamino) propan-1-ol, 3.4 g of hexane acid, 1 04 g of dicyclohexylcarbodiimide and 640 mg of benzotriazol-1-ol, analogously to example II f). A product with a molecular weight of 524.6 is obtained. (C ₃₁ H ₃₂ N ₄ O ₄ ); MS (ESI +): 525 (M + H ⁺ ): (ESI-): 523 (MH ⁺ ).

b) 4- {1- (4-Fluorophenyl) -3- [3- (4-fluorophenyl) -3-hydroxypropyl] -4-oxoazetidin-2-yl} benzylamide {2- [3-hydroxy-3-phenyl- 2-pyridin-2-yl-1- (pyridin-2-ylamino) propyl] phenyl} hexane acid amide ( 14 )

The product is obtained from 300 mg of 4- (4-aminomethylphenyl) -1- (4-fluorophenyl) -3- [3- (4-fluorophenyl) -3-hydroxypropyl] azetidin-2-one, 372 mg 5- {2- [ 3-hydroxy-3-phenyl-2-pyridin-2-yl-1- (pyridin-2-ylamino) propyl] phenylcarbamoyl} pentanoic acid, 155 mg of dicyclohexylcarbodiimide and 120 mg of benzotriazole-1-ol in 25 ml of methahydrofuran analogously to example III chromatography: SiO ₂ , dichloromethane / methanol = 10: 1; the molecular weight of the product is 929.1. (C ₅₆ H ₅₄ F ₂ N ₆ O ₅ ); MS (ESI +): 929 (M + H ⁺ ).

Example XIII

(1- {2- [8- (4- {1- (4-Fluorophenyl) -3- [3- (4-fluorophenyl) -3-hydroxypropyl] -4-oxoazetidin-2-yl} benzylcarbamoyl) octanoylamino] phenyl } -3-hydroxy-3-phenyl-2-pyridin-2-ylpropyl) pyridin-2-ylammonium trifluoroacetate ( 27 )

a) 8- {2- [3-Hydroxy-3-phenyl-2-pyridin-2-yl-1- (pyridin-2-ylamino) propyl] phenylcarbamoyl} octanoic acid ( 26 )

C ₃₄ H ₃₈ N ₄ O ₄ (566.72) MS (ESI) 567 (M + H).

b) (1- {2- [8- (4- {1- (4-Fluorophenyl) -3- [3- (4-fluorophenyl) -3-hydroxypropyl] -4-oxoazetidin-2-yl} benzylcarbamoyl) octanoylamino ] phenyl} -3-hydroxy-3-phenyl-2-pyridin-2-ylpropyl) pyridin-2-ylammonium trifluoroacetate ( 27 ).

C ₆₁ H ₆₁ F ₅ N ₆ O ₇ (1085.19) MS (ESI) 971 (M + H).

Example XIV

(1- {2- [8- (3- {1- (4-Fluorophenyl) -3- [3- (4-fluorophenyl) -3-hydroxypropyl] -4-oxoazetidin-2-yl} benzylcarbamoyl) octanoylamino] phenyl } -3-hydroxy-3-phenyl-2-pyridin-2-ylpropyl) pyridin-2-ylammonium trifluoroacetate ( 28 )

C ₆₁ H ₆₁ F ₅ N ₆ O ₇ (1085.19) MS (ESI) 971 (M + H).

Example XV

(1- {2- [11- (4- {1- (4-Fluorophenyl) -3- [3- (4-fluorophenyl) -3-hydroxypropyl] -4-oxoazetidin-2-yl} benzylcarbamoyl) undecanoylamino] phenyl } -3-hydroxy-3-phenyl-2-pyridin-2-ylpropyl) pyridin-2-ylammonium trifluoroacetate ( 30 )

a) 11- {2- [3-Hydroxy-3-phenyl-2-pyridin-2-yl-1- (pyridin-2-ylamino) propyl] phenylcarbamoyl} undecanoic acid ( 29 )

C ₃₇ H ₄₄ N ₄ O ₄ (608.82) MS (ESI) 609 (M + H).

b) (1- {2- [11- (4- {1- (4-Fluorophenyl) -3- [3- (4-fluorophenyl) -3-hydroxypropyl] -4-oxoazetidin-2-yl} benzylcarbamoyl) undecanoylamino ] phenyl} -3-hydroxy-3-phenyl-2-pyridin-2-ylpropyl) pyridin-2-ylammonium trifluoroacetate ( 30 )

C ₆₄ H ₆₇ F ₅ N ₆ O ₇ (1127.28) MS (ESI) 1013 (M + H).

Example XVI

(1- {2- [11- (3- {1- (4-Fluorophenyl) -3- [3- (4-fluorophenyl) -3-hydroxypropyl] -4-oxoazetidin-2-yl} benzylcarbamoyl) undecanoylamino] phenyl } -3-hydroxy-3-phenyl-2-pyridin-2-ylpropyl) pyridin-2-ylammonium trifluoroacetate ( 31 )

C ₆₄ H ₆₇ F ₅ N ₆ O ₇ (1127.28) MS (ESI) 1013 (M + H).

Example XVII

(1- {2- [2- (2- {2 - [(4- {1- (4-Fluorophenyl) -3- [3- (4-fluorophenyl) -3-hydroxypropyl] -4-oxoazetidine-2- yl} benzylcarbamoyl) methoxy] ethoxy} ethoxy) acetylamino] phenyl} -3-hydroxy-3-phenyl-2-pyridin-2-ylpropyl) pyridin-2-ylammonium trifluoroacetate ( 33 )

a) {2- [2 - ({2- [3-Hydroxy-3-phenyl-2-pyridin-2-yl-1- (pyridin-2-ylamino) propyl] phenylcarbamoyl} methoxy) ethoxy] ethoxy} acetic acid ( 32 )

C ₃₃ H ₃₆ N ₄ O ₇ (600.68) MS (ESI) 591 (M + H).

b) (1- {2- [2- (2- {2 - [(4- {1- (4-Fluorophenyl) -3- [3- (4-fluorophenyl) -3-hydroxypropyl] -4-oxoazetidine- 2-yl} benzylcarbamoyl) methoxy] ethoxy} ethoxy) acetylamino] phenyl} -3-hydroxy-3-phenyl-2-pyridin-2-ylpropyl) pyridin-2-ylammonium trifluoroacetate ( 33 )

C ₆₀ H ₅₉ F ₅ N ₆ O ₁₀ (1119.17) MS (ESI) 1005 (M + H).

Example XVIII

(1- {2- [2- (2- {2 - [(3- {1- (4-Fluorophenyl) -3- [3- (4-fluorophenyl) -3-hydroxypropyl] -4-oxoazetidine-2- yl} benzylcarbamoyl) methoxy] ethoxy} ethoxy) acetylamino] phenyl} -3-hydroxy-3-phenyl-2-pyridin-2-ylpropyl) pyridin-2-ylammonium trifluoroacetate ( 34 )

C ₆₀ H ₅₉ F ₅ N ₆ O ₁₀ (1119.17) MS (ESI) 1005 (M + H).

Example XIX

(1- [2- (2- {2 - [(4- {1- (4-Fluorophenyl) -3- [3- (4-fluorophenyl) -3-hydroxypropyl] -4-oxoazetidin-2-yl} benzylcarbamoyl ) methoxy] ethoxy} acetylamino) phenyl] -3-hydroxy-3-phenyl-2-pyridin-2-ylpropyl} pyridin-2-ylammonium trifluoroacetate ( 36 )

C ₃₁ H ₃₂ N ₄ O ₆ (556.62) MS (ESI) 557 (M + H).

a) 2 - ({2- [3-Hydroxy-3-phenyl-2-pyridin-2-yl-1- (pyridin-2-ylamino) propyl] phenylcarbamoyl} methoxy) ethoxy] acetic acid ( 35 )

b) {1- [2- (2- {2 - [(4- {1- (4-Fluorophenyl) -3- [3- (4-fluorophenyl) -3-hydroxypropyl] -4-oxoazetidin-2-yl } benzylcarbamoyl) methoxy] ethoxy} acetylamino) phenyl] -3-hydroxy-3-phenyl-2-pyridin-2-ylpropyl} pyridin-2-ylammonium trifluoroacetate ( 36 )

C ₅₈ H ₅₅ F ₅ N ₆ O ₉ (1075.11) MS (ESI) 961 (M + H).

Example XX

{1- [2- (2- {2 - [(3- {1- (4-Fluorophenyl) -3- [3- (4-fluorophenyl) -3-hydroxypropyl] -4-oxoazetidin-2-yl} benzylcarbamoyl ) methoxy] ethoxy} acetylamino) phenyl] -3-hydroxy-3-phenyl-2-pyridin-2-ylpropyl} pyridin-2-ylammonium trifluoroacetate ( 37 )

C ₅₈ H ₅₅ F ₅ N ₆ O ₉ (1075.11) MS (ESI) 961 (M + H).

The activity of the compounds of formula I according to the invention was tested by the methods described below.

Effect of cholesterol absorption + ³ H-excretion of taurocholic acid by fecal excretion of mice, rats or hamsters

NMRI mice, Wistar rats, or Golden Syrian hamsters (in groups n = 4-6) were kept on a standard diet (altromin, Lage (Lippe)) in cells. In the afternoon, before giving the radioactive labeled atoms ( ¹⁴ C-cholesterol), the animals were kept on an empty stomach and adapted on the grate.

An additional 24 hours before oral administration of the control feed ( ¹⁴ C-cholesterol in Intralipid® 20, Pharmacia-Upjohn) the animals were labeled with ³ H-TCA (taurocholic acid) (for example, 1 μCi for mouse up to 5 μCi for rat).

Cholesterol absorption test: 0.25 ml of Intralipid® 20 (Pharmacia-Upjohn) (oral injection of 0.25 μCi ¹⁴ C-cholesterol in 0.1 mg of cholesterol) was administered orally to one mouse using a gastric tube.

Test substances were separately dissolved in 0.5% / (methyl cellulose (Sigma) / 5% Solutol (BASF, Ludwigshafen) or added to a suitable base.

The volume of test substance administered was 0.5 ml per mouse. The test substance was administered immediately before the control feed (Intralipid with labile ¹⁴ C-cholesterol) (cholesterol absorption test).

Feces were collected within 24 hours: fecal excretion after 24 hours was analyzed for ¹⁴ C-cholesterol and ³ N-taurocholic acid (TCA).

The liver was removed, homogenized, and an aliquot was burned in oximeate (Model 307, Packard) to determine the amount of ¹⁴ C-cholesterol absorbed / resorbed.

Evaluation of the results:

Samples of feces :

The total weight was determined, adjusted to a certain volume with water, then homogenized, an aliquot was dried and burned in an oximate (model 307, Packard for burning labeled radioactive samples): the amount of radioactive ³ H-H ₂ O and ¹⁴ C-CO _{2 were} calculated on the allocated amount ³ N-taurocholic acid or ¹⁴ C-cholesterol (binary isotope method). The ED ₂₀₀ values were interpolated from the dose efficiency curve as a dose that doubles the release of TCA or cholesterol, based on the simultaneously treated control group.

Liver tests:

The absorbed amount of ¹⁴ C-cholesterol in the liver was converted to the administered dose. The ED ₅₀ values were interpolated from the dose efficiency curve as a dose that halves (50%) the absorption of ¹⁴ C-cholesterol in the liver in terms of the control group.

The following ED ₅₀ values prove the activity of the compounds of formula I according to the invention

Example ED ₅₀ (liver) [mg / mouse] XVI 0,03 XVIII 0.3 XIX 0.1

From the table it can be seen that the compounds of formula I have a very good cholesterol lowering effect.

Suction

The absorption of the compounds of formula I was tested on a Saso-cell model (A.R. Hilgers et al., Caco-2 cell monolayers as a model for drug transport across the intestinal mucosa, Pharm. Res. 1990, 7, 902).

From the above data it can be seen that the compounds of formula I according to the invention, compared with the compounds known from the prior art (comparison structure), exhibit a significantly lower resorption.

Known Compound Example Apparent separation coefficient P _arr [cm / sec] (according to Hilgers) 4.88CH10 ^-06

Estimated human resorption 100%

Known Compound: Ezetimibe

Claims (4)

1. The compounds of formula I

wherein R1 is hydrogen; R2 means - (CH ₂ ) _0-1- NH-C (O) - (C ₃ -C ₁₂ ) alkylene-C (O) NH-L, where one or more C-atoms in the alkylene residue may be substituted with an O-atom; R3 and R6 are independently hydrogen, R4 and R5 are independently F, Cl, Br, I,

Rx, Ry, Rz mean H, and their pharmaceutically acceptable salts. 2. A pharmaceutical composition exhibiting cholesterol lowering activity and containing one or more compounds according to paragraph 1. 3. A method of obtaining a pharmaceutical composition containing one or more compounds according to claim 1, characterized in that the active substance is mixed with a pharmaceutically acceptable carrier and this mixture is converted into a usable form. 4. The use of compounds according to claim 1 for the preparation of a pharmaceutical composition exhibiting cholesterol lowering activity.