AU667009B2

AU667009B2 - Bile acid derivatives, processes for their preparation and the use of these compounds as medicaments

Info

Publication number: AU667009B2
Application number: AU61948/94A
Authority: AU
Inventors: Alfons Enhsen; Heiner Glombik; Werner Kramer; Gunther Wess
Original assignee: Hoechst AG
Current assignee: Hoechst AG
Priority date: 1993-05-08
Filing date: 1994-05-06
Publication date: 1996-02-29
Anticipated expiration: 2014-05-06
Also published as: NO941679L; HUT67522A; CZ113694A3; FI942076A0; FI942076A; AU6194894A; EP0624593A3; NZ260470A; NO941679D0; HU9401411D0; CA2123050A1; EP0624593A2; JPH07304792A; IL109579A0

Description

N

Regulation 3.2(2)

AUSTRALIA

Patents Act 1990 6 00

ORIGINAL

COMPLETE SPECIFICATION STANDARD PATENT

S

a a. a Application Number: Lodged: Invention Title: BILE ACID DERIVATIVES, PROCESSES FOR THEIR PREPARATION AND THE USE OF THESE COMPOUNDS AS MEDICAMENTS a a, The following statement is a full description of this invention, including the best method of performing it known to us HOECHST AKTIENGESELLSCHAFT HOE 93/F 131 Dr.D/sch Description Bile acid derivatives, processes for their preparation and the use of these compounds as medicaments The invention relates to novel bile acid derivatives, processes for their preparation, pharmaceutical preparations based on these compounds and the use of the bile acid derivatives as medicaments.

Bile acids have an important physiological function in lipolysis, for example as cofactors of pancreatic lipases and as natural detergents for solubilizing fats and fatsoluble vitamins. As the end product of cholesterol metabolism, they are synthesized in the liver, stored in the gall bladder and secreted from this by contraction into the small intestine, where they display their physiological action. The greatest proportion of the bile acids secreted is recovered via the enterohepatic circulation. They return to the liver via the mesenterial :veins of the small intestine and the portal vein system.

20 Both active and passive transportation processes play a role in reabsorption in the intestine. Most of the bile acids is reabsorbed at the end of the small intestine, the terminal ileum, by a specific Na-dependent transportation system, and returns to the liver with the mesenterial vein blood via the portal vein, to be secreted by the liver cells again into the bile. The bile acids appear in the enterohepatic circulation both as free acids and in the form of glycine conjugates and taurine conjugates.

Non-absorbable, insoluble, basic, crosslinked polymers have been used for many years for binding bile acids and utilized therapeutically on the basis of these properties. Bile acid derivatives described in Patent Application EP-A-0 489 423 have a high affinity for the i L -e 2 intestinal bile acid transportation system and therefore allow specific inhibition of the enterohepatic circulation. All diseases in which inhibition of bile acid resorption in the intestine, in particular in the small intestine, seems desirable are regarded as the therapeutic object. For example, the biligenic diarrhea following ileum resection or increased blood cholesterol levels are treated in this manner. In the case of increased blood cholesterol level, a reduction in this level can be achieved by intervention in the enterohepatic circulation. The corresponding new synthesis of bile acids from cholesterol in the liver is caused by lowering the bile acid pool in the enterohepatic circulation. The LDL-cholesterol in the blood circulation is resorted to in order to meet the cholesterol requirement in the liver, the hepatic LDL receptors increasingly being used. The acceleration of LDL metabolism which has thus occurred takes effect by reducing the atherogenic cholesterol content in the 20 blood.

The object was to discover novel medicaments which are capable of reducing the atherogenic cholesterol content in the blood or of influencing the enterohepatic circulation in respect of increased excretion of bile acid and consequent reducticn in the cholesterol level.

This object is achieved by the bile acid derivatives according to the invention.

EP-A-0 489 423 relates to dimeric bile acid derivatives of the formula .:oo 30 G1-X-G2 in which Gl and G2 are linked in positions 3, 7 or 12 or by the side chain via the linker X. Bile acid derivatives in which G1 is bonded to X via positions 7 or 12 and G2 is bonded to X via positions 3, 7 or 12 or the side chain LslY IIII 3 are not described in the examples of the European Patent Application cited.

The invention therefore relates to bile acid derivatives of the formula I G1 X G2 I in which G1 is a radical of the formula II

R

4 H R in which Y has the following meaning: OKa, in which Ka is an *o alkali metal, alkaline earth metal or quaternary 1 0 ammonium ion, -OL, -NHL, -NL 2 an amino acid or aminosulfonic acid bonded via the amino group, such as, for example

-NHCH

2 COOH, -NHCH 2 CH SO 3 H, -NCHgCOOH, -NCH 2

CH

2

SO

3

H

:I

I

CH

3 CH 3 thereof, and in which L is H, an alkyl or alkenyl radical having up to carbon atoms, which is branched or unbranched, a cycloalkyl radical having 3 to 8 carbon atoms or a phenyl or benzyl radical, which are unsubstituted or mono- to trisubstituted by F, Cl, Br, (Ci-C 4 )-alkyl

_I

4 or (Cz-C 4 )-alkoxy,

R

1 is H, an alkyl or alkenyl rae'.cal having up to carbon atoms, which is branched or unbranched, a cycloalkyl radical having 3 to 8 carbon atoms, a benzyl radical, a biphenylmethyl or a triphenylmethyl radical, in which the nuclei are unsubstituted or mono- to trisubstituted by F, Cl, Br, (Cz-C 4 )-alkyl or

(C,-C

4 )-alkoxy, or a radical o 0 I 11 0 -P-OL, -S-OL or f 11 II

-C-L

0 0 in which L has the abovementioned meaning,

R

2 to R 5

R

2 and R 3 or R 4 and R 5 in each case together being the oxygen of a carbonyl group, or indi- 15 vidually and in each case independently of one another being 0 0 0 OL 0 :II II II I II H, -OT, -ST, -NHT, O-C-T, -NH-C-T -OS-T, -T I II 0 0 in which T has the meaning of L or is a free valency for bonding the group X, and in which y one free valency starts 20 from G1 for bonding the group X, X is a single bond or a group of the formula III 0 0

CH

2 )qC (111), I I I L L L 3 in which

~I~

5 A and B are alkylene chains, which are branched or unbranched, it being possible for the chains to be optionally interrupted by or L, L 2 and L' are identical or different and have the meaning of L and q is zero to r is zero or 1, s is zero or 1 and t is zero or 1 and G2 is a radical of the formula IV R' Rio S Iv) H H in which Z is a free valency to the group X or has the meaning given under Y,

R

6 is a free valency to the group X or has the meaning 15 given under R 1 and

R

7 to R" 1 have the meaning given under R 2 to R 5 and in which a=by one free valency starts from G2 to the group X.

The compounds according to the invention have a high affinity for the specific bile acid transportation system of the small intestine and inhibit bile acid absorption in a concentration-dependent and competitive manner. The compounds according to the invention furthermore are not themselves absorbed and thus do not enter the blood circulation. The enterohepatic circulation can be interrupted very specifically and efficiently by application of this principle of action.

By using the compounds according to the invention, it is possible to reduce the amount of bile acids in the 6 enterohepatic circulation such that a reduction of the cholesterol level in the serum occurs. Avitaminoses are just as unlikely during their use as effects on the absorption of other medicaments or an adverse effect on the intestinal flora. Furthermore, the side-effects known of polymers (constipation, steratorrhea) are not found, i.e. lipolysis is not adversely influenced. Because of the high affinity for the specific bile acid transportation system of the small intestine, low daily doses are sufficient, so that acceptance of such medicaments by the doctor and patient will be very high.

Particularly preferred compounds of the formula I are those in which G1 is a radical of the formula II

R

4 H Df Y

*H

in which G ot Y OH, 0-(C 1

-C

4 )-AlkyI, -NHCH 2

COOH,

-NCH

2 COOH, -NHCH 2

CH

2

SO

3 H, -NCH 2

CH

2

SO

3

H

CH

3

CH

3 15 R' is H, benzyl, biphenylmethyl, formyl or acetyl,

R

2 to R 5

R

2 and R 3 or R 4 and R 5 in each case together being the oxygen of a carbonyl group, or individually and in each case independently of one another being o 0 II 11 H, -OT, -NHT, -NH-C-T, -T in which T is H, a branched or unbranched (Ci-C 4 )-alkyl radical or L I I II 7 a free valency to bridge group X, and in which a.

t&=bah e one free valency starts from G1 for bonding the group X, X is a bond,

H

-CH

2

CH

2

NH-

-CH

2

CH

2

CH

2

NH-

0 0 II II

-(CH

2 )n-N-C-(CH 2 )m-C-N-(CH 2 o H H .o s 1

I

s where n is 2 or G2 is a radical of 3, m is 1 to 4 and o is 2 or 3, and the formula IV R9 RI 0 0 e eo *0 0 oe eeoc eeee I V in which Z is a free valency to group X or hac the meaning given above under Y,

R

6 is a free valency to group X or has the meaning given above under R 1 and

R

7 to R 10 have the meaning given above under R 2 to R s and in which only one free valency starts from G2 to the group X.

The invention furthermore relates to a process for the Spreparation of compounds of the formula I, which 8 comprises a) in the case where X is a single bond, reacting suitable forms of Gl and G2 with one another by processes which are known in principle, or b) in the case where X is a bridge group, reacting a) reactive forms of G1-X with G2 or P) reactive forms of G2-X with G1 by processes which are known in principle, or c) preparing compounds of the formula I (G1-X-G2) from G1-Xl and X2-G2 by processes which are known or, where they are not known, by the processes described below in more detail, X being formed from Xl and X2 by formation of a covalent bond, in particular within a condensation or substitution reaction.

S 15 a) X is a single bond *.The bile acids Gl are employed either in the free o: form or in protected form. After linking with G2, which is likewise present in a free or protected form, the protective groups are split off, if appropriate, and the C-24 carboxyl function is converted into a derivative, if appropriate. Suitable protective groups for the alcohol groups are expediently formyl, acetyl, tetrahydropyranyl or t-butyldimethylsilyl. Various alkyl or benzyl 25 esters, and also, for example, orthoesters, are suitable protective groups for the C-24 carboxyl group.

For example, bile acid preferentially reacts at position 3, but also at position 7, with activated forms of carboxylic acids, such as acid chlorides or mixed anhydrides, with addition of bases, such as trialkylamire or pyridine, but also NaOH, at room temperature in suitable solvents, such as tetrahydrofuran, iethylene chloride or ethyl acetate, but also dimethylformamide (DMF) or dimethoxyethane

(DME).

9 The various isomers can be separated, for example by chromatography. The reaction can be carried out selectively by using suitable protective groups.

The corresponding amino-bile acids can be converted into corresponding amides analogously. Here also, the reaction can be carried out either with protected or with free bile acids.

Other compounds according to the invention can be linked analogously by known standard processes.

b) X is a bridge group The processes specified under a) are also used to carry out the linking of G1-X with G2 or G1 with X-G2. Here also, the bile acid portion is expediently employed either in protected or in 15 unprotected form.

A preferred preparation process comprises reacting reactive forms of G1 with reactive forms of X-G2. If appropriate, the linking reaction is followed by splitting-off of protective groups and conversion of C-24 carboxyl into derivatives.

The preparation of reactive bile acid units Gl-X and X-G2 is shown in the following equation.

0 OH

H

(V)

10 0 'o me H H C H2 0

H

v I H 2

H

H He R 0 RI *ei* 0* *0 0

H

R 0\o 00 I I I I I x) 11 H0 (x xI R H, formyl or acetyl, R' H or OH, R" formy. or acetyl a.

VHI0 (X I I) (XI I I a 12 X or X II R 0 (xIv) 0- (CH 2

-NH

2 e a H 0b X I or XI I I H2N- CH2)O nO H 0/ OMe \H H R 0 R

H

RO

(XV)

DT-r- N- CH2),I C ou Hl

D

aeo e r s so o o

D

ro n o (xv I)

(XVII)

R H, formyl or acetyl, R' H or OH, n 2 or 3 Compounds of the type V in which the 3-position is protected are reacted with allyl bromide/HUnig base or triethylamine. If the compound V has one OH group, the alkylation is unambiguous; if two free OH groups are present, monoalkylation takes place at positions 7 and 12 in approximately equal proportions and only traces of the dialkylated product are formed. The protective group in the 3-position can either be split off with sodium methylate or retained for further reactions. The -I I~LJII- ~I 13 monoalkylated compounds VI and VII can be split with ozone or with OsO 4 /NaIO 4 to give the aldehydes VIII and IX. The 7-and 12-hydroxyethyl compounds X and XI are readily accessible from these by simple reduction, for example with NaBH 4 The corresponding 7- and 12-hydroxypropyl derivatives XII and XIII can be synthesized from the allyl compounds VI and VII by hydroboration. The aminoalkyl derivatives XIV and XV can be prepared from the hydroxyalkyl compounds of the type X to XIII by a reaction sequence which is known in principle (mesylation of the primary OH group with methanesulfonyl chloride/ pyridine, azide exchange with NaN 3 in dir.ethylformamide, reduction of the azide function with hydrogen under catalytic conditions). Further reaction of the amino 15 functions of these compounds with succinic anhydride Sgives bile acid units of the type XVI and XVII. Suitable bile acid units furthermore are described in EP-A- 0 489 423.

The invention furthermore relates to the use of the compounds according to the invention for the preparation of a medicine. For this, the compounds of the formula I are dissolvel or suspended in pharmacologically acceptable organic solvents, such as mono- or polyhydric "alcohols, such as, for example, ethanol or glycerol, or 25 in triacetin, oils, such as, for example, sunflower oil or cod-liver oil, ethers, such as, for example, diethyl- 'ene glycol dimethyl ether, or also polyathers, such as, oi' for example, polyethylene glycol, or also in the presence of other pharmacologically acceptable polymeric carriers, such as, for example, polyvinylpyrrolidone, or other pharmaceutically acceptable additives, such as starch, cyclodextrin or polysaccharides. The compounds according to the invention furthermore can be administered in combination with other medicaments.

The compounds of the formula I are administered in various dosage forms, preferably orally in the form of tablets, capsules or liquids. The daily dose varies in I~13 14 the range from 3 mg to 5000 mg, but preferably in the dose range from 10 to 1000 mg, depending on the body weight and constitution of the patient.

On the basis of their pharmacological properties, the compounds are particularly suitable as hypolipidemic agents.

The invention therefore also relates to medicaments based on the compounds of the formula and to the use of the compounds as medicaments, in particular for lowering the cholesterol level.

The compounds according to the invention were tested biologically by determination of the inhibition of 3

H]-

taurocholate uptake in brush border membrane vesicles of the ileum in rabbits. The inhibition test was carried out 15 as follows: 1. Preparation of brush border membrane vesicles from the ileum of rabbits The brush border membrane vesicles from the intestinal cells of the small intestine were prepared by the 20 so-called Mg 2 precipitation method. Male New Zealand rabbits (2 to 2.5 kg body weight) were sacrificed by intravenous injection of 0.5 ml of an aqueous solution of 2.5 mg of tetracaine HC1, 100 T 61" and 25 mg of mebezonium iodide. The small intestine was removed and flushed with ice-cold physiological saline solution. The terminal 7/10 of the small intestine (measured in the oral-rectal direction, i.e. the terminal ileum which contains the active Na+-dependent bile acid transportation system) was used for preparation of the brush border membrane vesicles. The intestines were frozen in plastic bags under nitrogen at -80°C. To prepare the membrane vesicles, the frozen intestines were thawed at in a water-bath. The mucosa was scraped off and suspended in 60 ml of ice-cold 12 mM Tris/HCl buffer (pH 15 7.1)/300 mM mannitol, 5 mM EGTA/10 mg/l of phenylmethylsulfonyl fluoride/1 mg/l of trypsin inhibitor from soybeans (32 U/mg)/0.5 mg/l of trypsin inhibitor from bovine lung (193 U/mg)/5 mg/l of bacitracin. After dilution to 300 ml with ice-cold distilled water, the mixture was homogenized with an Ultraturrax (18-rod, IKA Werk Staufen, FRG) for 3 m.'nutes at 75% of the maximum output, while cooling with ice. After addition of 3 ml of 1 M MgCl 2 solution (final concentration 10 mM), the mixture was left to stand at 0 C for exactly 1 minute. By addition of Mg 2 the cell, membranes aggregate and precipitate with the exception of the brush border membranes. After centrifugation at 3000 x g (5000 rpm, SS-34 rotor) for 15 minutes, the precipitate is discarded and the supernatant, which contains the brush border membranes, is centrifuged at 267000 x g (15000 rpm, SS-34 ":rotor) for 30 minutes. The supernatant was discarded and the precipitate was rehomogenized in 60 ml of 12 mM Tris/HC1 buffer (pH 7.1)/60 mM mannitol, 5 mM EGTA using a Potter Elvejhem homogenizer (Braun, Melsungen, 900 rpm, strokes). After addition of 0.1 ml of 1 M MgCl 2 solution and an incubation time of 15 minutes at 0°C, the S. mixture was centrifuged again at 3000 x g for 15 minutes.

The supernatant was then centrifuged again at 46000 x g 25 (15000 rpm, SS-34 rotor) for 30 minutes. The precipitate was taken up in 30 ml of 10 mM Tris/Hepes buffer (pH 7.4)/300 mM mannitol and resuspended homogeneously by strokes in a Potter Elvejhem homogenizer at 1000 rpm.

After centrifugation at 48000 x g (20000 rpm, SS-34 rotor) for 30 minutes, the precipitate was taken up in to 2 ml of Tris/Hepes buffer (pH 7.4)/280 mM mannitol (final concentration 20 mg/ml) and resuspended with the aid of a tuberculin syringe with a 27 gauge needle. The vesicles were either used for transportation studies immediately after preparation or stored in 4 mg portions in liquid nitrogen at -196 0

C.

2. Inhibition of Na-dependent uptake of 3 H]-taurocholate in the brush border membrane vesicles of the 16 ileum The uptake of substrates in the brush border membrane vesicles described above was determined by means of the so-called membrane filtration technique. 10 pl of the vesicle suspension (100 pg of protein) were pipetted as drops onto the wall of a polystyrene incubation tube (11 x 70 mm) which contained the incubation medium with the corresponding ligands (90 pl). The incubation medium contained 0.75 pl 0.75 pCi 3 H(G)]-taurocholate (specific activity: 2.1 Ci/mmol)/0.5 pV of 10 mM taurocholate/8.75 pl of sodium transportation buffer (10 mM Tris/Hepes (pH 7.4)/100 M mannitol/100 mM NaCI) (Na-T-P) or 8.75 il of potassium transportation buffer (10 mM o Tris/Hepes (pH 7.4)/100 mM mannitol/100 mM KC1) (K-T-P) 15 and 80 il of the inhibitor solution in question as a solution in Na-T buffer or K-T buffer, depending on the experiment. The incubation medium was filtered through a polyvinylidene fluoride membrane filter (SYHV LO 4NS, 0.45 pm, 4 mm 0, Millipore, Eschborn, FRG). The transportion measurement was started by mixing the vesicles with the incubation medium. The concentration of taurocholate in the incubation batch was 50 iM. After the desired incubation time (usually 1 minute), the transportation was stopped by addition of 1 ml of ice-cold stopping 25 solution (10 mM Tris/Hepes (pH 7.4)/150 mM KC1).

The mixture formed was immediately filtered off with suction over a membrane filter of cellulose nitrate (ME 0.45 pm, 25 mm diameter, Schleicher Schuell, Dassell, FRG) under a vacuum of 25 to 35 mbar. The filter was rinsed with 5 ml of ice-cold stopping solution.

To measure the uptake of radioactively labeled taurocholate, the membrane filter was dissolved with 4 ml of the scintillator Quickszint 361 (Zinsser Analytik GmbH, Frankfurt, FRG) and the radioactivity was measured by liquid scintillation measurement in a TriCarb 2500 measuring instrument (Canberra Packard GmbH, Frankfurt, 17 FRG). After calibration of the instrument with the aid of standard samples and after correction for any chemiluminescence present, the values measured were obtained as dpm (decompositions per minute).

The control values were determined in each case in Na-T-P and K-T-P, The difference between the uptake in Na-T-P and K-T-P gave the Na+-dependent transportation content.

The concentration of inhibitor at which the Na+-dependent transportation content was inhibited by 50% based on the control was designated the IC.

0 Na'.

The pharmacological data include a test series in which the interaction of the compounds according to the invention with the intestinal bile acid transportation system in the terminal small intestine was investigated. The oe« 15 results are summarized in Table 11.

S

Examples 1 and 2 *fee H N o~ 0W, HO H M Ac H Ace, H H H H Ac f oHI'H .ey o c i 'B 1 esp. 2 o*up. i *r 150 g (0.32 mol) of methyl 3-acetyl-cholate, 500 ml of off* 0 *dimethylformamide, 125 ml of N-ethyl-diisopropylamine and ml of allyl bromide are heated under reflux for 16 hours. New allyl bromide (25 ml) is added every 2 hours.

The reaction solution is evaporated on a rotary evaporator. The residue is partitioned between water/methylene chloride and the organic phase is separated off and dried with magnesium sulfate. After column chromatography (ethyl acetate/cyclohexane 1:2, silica gel 70-200 pm), the product fractions are evaporated on a rotary evaporator.

Yield 92.2 g of 7-/12-allyl mixture.

~n 18

C

30

H

4 a0 6 (504), MS 511 (M Li The mixture was separated by fractional crystallization with n-heptane.

Example 3 'OMe Ac0 50 g (0.1 mol) of Example 1, 250 ml of diethyl ether and 250 ml of water are initially introduced into the reaction vessel, while stirring vigorously. 503 mg (0.002 mol) of osmium tetroxide are added. The mixture is stirred at room temperature for 15 minutes. 53 g (0.25 mol) of sodium periodate are added in portions over the course of 1 hour, and the mixture is subsequently stirred for 8 hours, while stirring vigorously. The ether phase is separated off, dried with magnesium sulfate and evaporated on a rotary evaporator.

Yield: 47 g of crude C 29

H

46 0 7 (506), MS 513 (M Li') Example 3 is further reacted without additional purification.

Example 4 H0

OM'

H

Ac 0 0 SH0

H

4.2 g (0.11 mol) of sodium borohydride are added in portions to 47 g (0.093 mol) of Example 3 and 250 ml of I 19 methanol at 0 C. After 2 hours at 0°C, the reaction solution is poured onto saturated ammonium chloride solution, the mixture is extracted 3 times with ethyl acetate and the combined organic phases are dried with magnesium sulfate and evaporated on a rotary evaporator.

After column chromatography (ethyl acetate/cyclohexane 1.5:1, silica gel 35 70 pm), the product fractions are evaporated on a rotary evaporator and the residue is crystallized with diisopropyl ether. Yield: 25 g of

C

29

H

48 0 7 (508), MS 515 (M Li Example HO HO H4

O

M

e H 0 OMe

HO

A H' Ac' aCIo 0'O0H

H

10 g (0.02 mol) of Example 1 and 250 ml of tetrahydrofuran were initially introduced into a reaction vessel at room temperature, and 40 ml (0.04 mol) of borane-tetra- 15 hydrofuran complex (1 molar) were added dropwise at room temperature. The mixture was subsequently stirred at room temperature for 2 hours, and 25 ml of water, 25 ml of 2 N sodium hydroxide solution and 25 ml of 35% strength hydrogen peroxide solution were added dropwise in succession. The mixture was subsequently stirred at room temperature for a further 15 minutes. The reaction solution was poured onto water, the mixture was extracted 3 times with diethyl ether and the combined organic S 5 phases were dried with magnesium sulfate and evaporated 25 on a rotary evaporator.

Yield: 8.5 g of C 30 Hs0 7 (522), MS 529 (M Li Example 5 was further reacted without additional purification.

20 Example 6 HO

HO

H oH OMe A c0o/.,0 H A c N- H 2 Ac~~ H 1 °0

H

g (0.02 mol) of Example 4 and 100 ml of pyridine are initially introduced into a reaction vessel at 0°C.

1.7 ml (0.022 mol) of methanesulfonyl chloride are added dropwise at 0°C and the mixture is subsequently stirred at 0°C for a further 30 minutes and at room temperature for 2 hours. The reaction solution is poured onto water, the mixture is extracted 3 times with ethyl acetate, and the combined organic phases are dried with magnesium sulfate and evaporated on a rotary evaporator. The residue is dissolved in 100 ml of dimethylformamide, 1.4 g (0.022 mol) of sodium azide are added and the mixture is stirred at 80 0 °C for 2 hours. The reaction solution is poured onto water and the mixture is worked 15 up as described above. The residue is dissolved in 100 ml of methanol, 100 mg of palladium-on-charcoal are added and hydrogenation is carried out under normal pressure for 2 hours. The catalyst is filtered off and the filtrate is evaporated on a rotary evaporator. After 20 column chromatography (ethyl acetate/ MeOH/Et 3 N 10:1:1, silica gel 70-200 im), Example 6 is obtained.

Yield 7.3 g of C 29

H

49

NO

6 (507), MS 514 (M Li Example 7 HO

HO

o* u O/ 0 A n H H H H H 0 L-L_ LI 21 98.6 mg (0.001 mol) of succinic anhydride are added to 500 mg (0.001 mol) of amino compound, 20 ml of tetrahydrofuran and 4 ml of triethylamine at room temperature.

The mixture is subsequently stirred at room temperature for 1 hour. The reaction solution is poured onto strength sodium dihydrogen phosphate solution, the mixture is extracted 3 times with ethyl acetate and the organic phase is dried with magnesium sulfate and evaporated on a rotary evaporator.

Yield: 580 mg of C 33

H

53 NOg (607), MS 614 (M Li') Example 7 was further reacted without additional purification.

Examples to 7.

8 to 12 were prepared analogously to Examples 3 Examples 8-12 o r o AcO o r s Example R" MS 8 -CH2CHO 513 (M Li 9 -CH 2

CH

2 0H 515 (M Li' -CHCHCHOH 529 (M Li 11 -CH 2

CH

2

NH

2 514 (M Li 12 -CH 2 CH2NHCOCH 2

CH

2 COOH 614 (M Li 22 Example 13 HO

HT

cholic ai -H aci

H

H H

HOO

300 mg ('0.73 mmol) of cholic acid, 330 mg (0.78 mmol) of methyl 7p-amino-3a, 12a-dihydroxy-50-'cholanate (Redel, Bull. Soc. Chim. Fr., page 877, 1949), 240 mg (0.97 mmol) of EEDQ and 0.25 ml of diisopropylethylamine are stirred in 20 ml of DMF at 90*C for 4 hours. After cooling, the reaction mixture is concentrated and the residue is chromatographed over silica gel (CH 2 Cl 2 /Me-OH C 49

H

81 N0 8 (812) 819 (M The two bile acid derivatives can also be linked with triethylamine in methylene chloride or with dicyclohexylcarbodiimide,, hydroxybenzotr,- Lzole or triethylamine in tetrahydrofuran.

The compounds of Table 1 were prepared analogously to Example 13.

Table 1

HO

H0 OMe H 13 14

RR

23 Example R'1 2 R 1 3

R

14 MS (FAB, 3-NBA/Licl) 14 ce-OH H -OH C 49

H

8 1

NO

7 (796) 803 (M Li+) P1-OH H -OH C 49 HqjNO 7 (796) 803 (M Li-) 16 H H -OCHO C 50

H

81 N0 7 (808.5) 809.5 The examples of Table 2 were obtained analogously to Example 13 from Examples 7 and 8.

.*1 .00.C &so*0 a a.

a.. a 0 *0 a a a. a a a a. a a a *a a a a 0.S**O* a~ a he 0t* P S Table 2 H0 Example V 3 MS (FAB, 3-NBA/LiCl) 17 -NH- H C 53

H

87 N0 10 (898) 905 (M Li-) 18 -NH- diphenylmethyl C 66 sH 97 N0 10 (1064) 1071 (M Li+) 19 -NHCO (CH 2 2 CQNH (CH 2 3 NH- H C 60

H

99

N

3 0 12 (1054) 1061 (M LiL+) -NHCO (CH 2 2 CONH (CH 2 3 NH- diphenylmethyl C 73

H

109

N

3 0 12 (1220) 1227 (M Li+) The examples of Tables 3 and 4 were likewise obtained analogously to Example 13.

Table 3 HO0 M e Ac H0 at0 as* Example R' 6 MS (FAB, 3-NBA/LiCl) 21 0 C 62

H

100

N

2 0 14 (1097) 1104

HO

22 C 6

H

58 2 0 1 (155)110 (MLe

HH

0 0 a *000, 00*0 Does 900* 26 Table 4

H

0000 *090 Example R 1 7 MS (FAB, 3-NBA/Lidl) 23 0 C 53

H

87 N0 10 (898) 905

-O

H 0*"0 H

H

24 0 C 62

H

10

N

2 0 14 (1097)1104 0 H

H

C

60

H

98 N0 13 (1055) 1062 0 1 j I 27 Example 26 H O 250 mg (0.31 mmol) of Example 13 are dissolved in 20 ml of ethanol, 2 ml of IN NaOH solution are added and the mixture is stirred at room temperature for 16 hours. For working up, the mixture is concentrated, the residue is dissolved in H 2 0, the pH is brought to 1-2 with 2N HC1 and the mixture is concentrated again. The residue is chromatographed over silica gel (CHC1 3 /MeOH 220 mg of free acid are obtained MS (FAB, 3-NBA/LiCl) C 4

,H,,NO

8 (798) 805 (M Li The examples of Tables 5 to 8 are obtained analogously to Example 26.

Table e• r oe o

H

R1 3

R

1 2

I

28 it..

6 .4

I.

0 0 Table 6 'q 0 c H 2~ c -X

R

1 Example VXR 3

MS

-NH- H C 5

,H

85 N0 10 (860) 867 (M Li-) 31 -NH- diphenylmethyl C 6

S

3

H.

95 N0 10 (1026) 1033 (M Li+) 32 -NHCO (CH 2 1-cOQNH (CH 2 3 NH- H C 5 7

H

9 7 N1 3 0 12 (1016) 1023 (M Li+) -NHCO (CH 2 2 CQNH (CH 2 3 NH- diphenylinethyl C 70

H

10

D

7

N

3 0 12 (1182) 1189 (M Li+) 30 Table 7 *9 Example R' 6

MS

34 HO 10 1 C 5 6

H

9 2 2 0 1 2 (985) 992 (M+Li+)

HOOH

H

31 Table 8 R17

H

HH

H t I/ H ai r r r r o a Example R 17 MS (FAB, 3-NBA/LiCl) 36 o C.

5

OH

3

ANO

9 (842) 849 (M+Li')

HO

H

37~tH

C

5 6

H

9 2 N0 1 2 (985) 992 (Ml+Li+) HO "0 H

H

38 0 C 56

H

9 2

N

2 0 1 2 (985) 992 (M+Li-)

OK

HH

0

H

The following glycine conjugates and taurine conjugates were obtained analogously to synthesis processeo which have already been described (EP 489 423).

0 0 Table 9 HO0

H

Example R 2R 3 MS (F,7BJ, 3-NBA/LiCI 39 a-OH ct-OH C 50

H

84

N

2 0 10 S (905) 918 (M 2Li+-H+) ct-OH Hi C 50

HS

4

N

2 0 9 S (889) 890 (M H+) 41 P-OH H C 5

,H

84

N

2 0 9 S (889) 912 (M Nae) 42 H C 50

H

84

N

2 0 8 S (872.5) 895.5 (M Na+) S S Table 0 3

H

CH

2 C H 2

-X

R 1 Example X3R1 5

MS

43 -NH- H C 52

]E

90

N

2 0 1 2 S (.967) 974 (M Li--) 44 -NH- diphenylmethyl C 6 5 Hl 00

N

2 0 12 S (1133) 1140 (M Li+) -NHCO (CH 2 2 CONH (CH 2 3 NH- B CS!A 10 2 N4 4 0 1 4 S (1123) 1130 (M Li+) 46 -NUCO (CH 2 2 CONH (CU 2 3 NH- diphenylmethyl C 72

HI

0 2

N

4 0 1 4 S (1289) 1296 (M Li+) 34 Example 47 H 0 3

H

MS (FAB, 3-NBA/LiCl) C 60

HIOIN

3 0 14 S (1092) 1099 (M Li+) Example 48 ,J e..

C.

P P

OC**

MS (FAB, 3-NBA/Lidl) C 72

H

110

N

4 0 13 (1239) 1246 (M Li+) Table 11 shows measurement values for the inhibition of the uptake of 3 H]-taurocholate in brush border membrane vesicles from the ileum of rabbits. The quotients of the and IC,( 0 Na values of the reference substance taurochenodeoxycholate (TCDC) and of the particular test substance are stated.

35 Table 11 Compound 1C 50 -TCDC ji.mol] IC 5 Ona-TCDC pmol] from Example substance pmol] IC50N- substance( mol] 0.00 0.12 26 0.00 0.29 27 0.64 0.44 28 0.54 0.43 29 0.23 0.17 0.93 0.85 32 1.00 0.80 39 0.92 1.05 40 0.54 0.52 43 1.18 0.96 470.35 0.26 48 0.75 0.71 a a.

S a.

a

Claims

2. A bile acid derivative of the formula I as claimed in claim 1, in which G1 is a radical of the formula II R 4 R K 0 n sc r r Pi Pi i e r o o r r 39 in which Y LS OH, O-(C 1 -C 4 )-Alkyl, .NHCH 2 COOH, -NCHCOH NCHCH S 3 H, .NCHZCH 2 S ZCOIHH S I S3 CH 3 C 3 RI is H, benzyl, biphenylmz':thyl, formyl or acetyl, R 2 to R 5 ,r R 2 and RI or RI and in each case together being the oxygen of a carbonyl group, or individually and in each case independently of one another being o 0 11 11 H, -OT, -NHT, -T in which T is 10 H, a branched or unbranched (Cl-C,)-alkyl radical or a free valency to bridge group X, and in which *a btai2. f one free valency starts from Gi for bonding the group X, X is a bond, H -CH2H2N* -CH 2 CH 2 CNH- 0 0 -(CH 2 ).CC 2 )C'C 2 )o 0 H H where n is 2 or 3, m is I to 4 and o is 2 or 3, and G2 is a radical of the formula IV I R9 H Z 0 H H (IV) R 8 H R7 in which Z is a free valency to group X or has the meaning given above under Y, R6 is a free valency to group X or has the meaning given above under R and R7 to Rio have the meaning given above under R2 to R5, and in which only one free valency starts from G2 to the group X.
3. A pharmaceutical comprising an effective amount of a bile acid derivative as claimed in claim 1 or claim 2 in adjunct with suitable carriers and/or excipients.
4. A hypolipidemic agent comprising an effective amount of a bile acid derivative as claimed in claim 1 or claim 2 in adjunct with suitable carriers S* and/or excipients. .0
5. A method of lowering cholesterol levels comprising administering to a patient requiring such treatment an effective amount of a bile acid derivative as claimed in claim 1. DATED this 12th day of December, 1995 HOECHST AKTIENGESELLSCHAFT WATERMARK PATENT TRADEMARK ATTORNEYS 290 BURWOOD ROAD HAWTHORN VICTORIA 3122 AUSTRALIA KJS/CJH/SH DOC 6 AU6194894.WPC c I 1 HOE 93/F 131 Abstract Bile acid derivatives, processes for their preparation and the use of these compounds as medicaments Bile acid derivatives of the formula I Gl X G2 in which X has the meaning given, G1 is a radical of the formula II (1 1) a 0@ 0. 0000 0 00 0.0. and G2 is a radical of the formula IV 0000 o0I 0 *o p Sr 0e*0 (iv) RO^ and in which the radicals RI to R 9 Y and Z occurring in the formulae II and IV have the meanings given, and processes for the preparation of these compounds are described. The bile acid derivatives of the formula I have useful pharmacological properties and can therefore be used as medicaments.