EP1786447A2

EP1786447A2 - Method for producing a ginger fraction and the use thereof for inhibiting human cyp enzymes

Info

Publication number: EP1786447A2
Application number: EP05776543A
Authority: EP
Inventors: Thomas Ebner; Eva Ludwig-Schwellinger; Stefan Matthias Blech
Original assignee: Boehringer Ingelheim International GmbH; Boehringer Ingelheim Pharma GmbH and Co KG
Current assignee: Boehringer Ingelheim International GmbH; Boehringer Ingelheim Pharma GmbH and Co KG
Priority date: 2004-08-28
Filing date: 2005-08-20
Publication date: 2007-05-23
Also published as: DE102004041716A1; WO2006024414A2; WO2006024414A3; JP2008511566A; US20060051437A1; CA2575797A1

Abstract

The invention relates to a method for producing a ginger fraction, to the fraction produced by using this method, and to its use alone or in conjunction with medicaments for inhibiting human cytochrome P450 (CYP) enzymes (in particular, cytochrome P450 3A4, CYP3A4) in order to positively influence the oral bioavailability and pharmacokinetics of active ingredients.

Description

Process for producing a ginger fraction and its use for inhibiting human CYP enzymes

The present invention is a process for producing a ginger fraction, the fraction produced by this process and their use alone or in combination with drugs for inhibiting human cytochrome P450 (CYP) enzymes (especially cytochrome P450 3A4, CYP3A4) to positively influence the oral bioavailability and pharmacokinetics of active substances.

Background of the invention

Cytochrome P450 (CYP) enzymes play a central role in drug metabolism. They are found mainly in the liver, but also in the intestinal wall, lungs, kidneys and other extrahepatic organs. Orally-dosed active ingredients may show only poor bioavailability due to the so-called "first-pass effect", for example those active substances which are susceptible to metabolism in the intestinal wall or the liver before the systemic blood circulation is reached. If the first-pass metabolism is inhibited, a marked increase in the bioavailability of orally dosed drugs can be achieved (Gibbs, Megan A. and Hosea, Natalie A .: Factors affecting the clinical development of cytochrome P450 3A substrates; Clin. Pharmacokinet 2003, 42 (11), 969-984). Many examples of drug-drug interactions that result in a higher bioavailability than the drug administered are based on such effects. Here, the first-pass metabolism of the drug is inhibited by another co-administered drug.

By inhibiting the first-pass metabolism, in addition to increasing the bioavailability of an active ingredient, the variability of the bioavailability, which according to experience increases with decreasing absolute bioavailability, can be significantly reduced. By reducing the variability, the bioavailability of the therapeutic success of oral drug therapy is significantly improved because it is less likely to occur too high (risk of unwanted side effects) or far too low (risk of treatment failure) drug exposure. Such effects may provide several benefits in drug therapy. For example, the HIV drug lopinavir shows inadequate bioavailability due to first-pass metabolism by CYP3A4. When given in a fixed-dose combination with ritonavir, which is a potent inhibitor of CYP3A4, significantly higher oral bioavailability of lopinavir is achieved.

However, the possibility of combining a drug with poor oral bioavailability with another drug or drug-like substance to reduce the first-pass effect is limited. This is mainly due to. the mode of action of the other active substance. Thus, e.g. For reasons of ethical reasons, an excessively low bioavailability of a cardiovascular drug is not increased by the simultaneous administration of an anti-retroviral active substance (indication HIV infection) to non-HIV-infected patients. become. Also, approved drugs are not registered for the purpose of inhibiting drug metabolizing enzymes. Instead, drug supplements can be helpful in this regard. For example, ingredients of grapefruit juice are potent inhibitors of CYP3A4 and drug transporters in the intestinal wall. There are numerous examples in the art which demonstrate that drug intake, along with grapefruit juice, has dramatic effects on the pharmacokinetics, safety and efficacy of orally-dosed drugs, such as, e.g. Simvastatin, cyclosporin A, terfenadine etc. (Ameer, Barbara and Weintraub, Randy A .: Drug interactions with grapefruit juice; Clin Pharmacokinet. 1997; 33 (2): 103-121).

Description of the invention

Ginger (Zingiber officinalis) is a common food ingredient in much of the world and is also used as a phyto-medicated for various purposes. For example, the powder of ginger root is available as a means of preventing seasickness. It contains about 5 to 8% of a viscous balsam (oleoresin), which contains a non-steam-volatile pungent fraction and a volatile volatile oil fraction.

The pale yellow essential oil occupies about 20 to 25% of the oleoresin. Depending on the origin of the composition of the essential oil is subject to strong fluctuations. It contains as main component sesquiterpene hydrocarbons of the bisabolone type, in particular (-) - α-zingiberene in addition to (-) - β-sesquiphellandrene, (-) - β-bisabolene, (+) - ar-curcumen and the acyclic α-farnesene (Deutsche Apothekerzeitung 1997, 137 (47), 40-46). The main component of the sharp fraction with a share of about 25% of the oleoresin is the homologous series of gingerols (HagerROM 2002: Zingiberis rhizoma, Springer Verlag, Heidelberg).

Surprisingly, in vitro experiments to inhibit CYP by various drugs and other compounds have shown that potent inhibition of various human CYP's can be achieved by a ginger fraction obtained by an extraction method of the present invention. This fraction shows a higher inhibitory potency (IC ₅₀ in the range below 1 μg / ml) both in comparison to the commercially available total ginger extract (the so-called oleoresin) and in comparison to the volatile fraction of the essential ginger oil (IC ₅₀ about 23 μg / ml), which is separated in the first extraction step.

The fraction obtained here is poorly soluble in hexane and differs by this property from the fraction of the essential oil, for the already inhibition of CYP3A4 has been shown (US 5,665,386).

The process according to the invention starts from a commercially available oleoresin and comprises several extraction steps with organic and aqueous solvents.

A first subject of the present invention is thus a process for the isolation of a ginger fraction with separation of the essential oil, comprising the steps

(a) extraction of an oleoresin with a non-polar organic solvent;

(b) extraction of the combined residues from step (a) with warm water and rejection of the supernatant. The residues thus obtained have an IC ₅₀ value of 0.9 μg / ml for CYP3A4. This value is obtained with the experiment described in the experimental part with human liver microsomes.

In a preferred embodiment, the residue thus obtained is further purified by a process comprising the steps

(c) extraction of the combined residues from step (b) with warm alcohol and

(d) concentrating the combined supernatants from step (c).

The fraction obtained in step (d) may be dissolved in an alcohol, preferably methanol or ethanol, and optionally further fractionated, for example by means of solid phase extraction and stepwise elution.

According to the invention, nonpolar organic solvents for use in step (a) are low-boiling alkane solvents such as hexane, heptane, octane, pentane or cyclohexane, petrochemical distillates, fuels and solvents such as, for example, gasoline, kerosene, petroleum ether, petroleum and other low-boiling, Volatile and non-polar solvents such as diethyl ether, ferric butyl methyl ether, tetrahydrofuran, benzene, toluene, xylene, with hexane is preferably used. The alcohol used in steps (c) and (e) may be selected from the group consisting of methanol, ethanol, isopropanol, n-propanol, n-butanol and other positionally isomeric butanols, n-pentanol and other positionally isomeric pentanols and may be the same or be different. Preferably, methanol is used. The extractant is used in each case in amounts of 4 to 10 ml / g, preferably 4 to 7 ml / g, of the oleoresin used.

The aqueous extractions are preferably carried out at a temperature of 50 to 80 ^° C., more preferably 65 to 75 ^° C.

As an alternative to this method, extractions with suitable aqueous organic acids or instead of a liquid-liquid extraction can also be used With organic solvents and solid phase extractions are used with suitable non-polar sorbents.

The extractions carried out in steps (a), (b) and (c) can be carried out one or more times, the phase comprising the desired product being combined from the various extractions of one working step. Preferably, the extraction of each operation is performed three times and the phases containing the product are combined. The combined phases are then processed further.

A second subject of the present invention is the ginger fraction according to the invention, obtainable by one of the processes according to the invention.

Preference is given to a ginger fraction which contains at least one compound of the general formulas

- (V)

wherein

n is the number 1, 2 or 3,

R ^{1 is} H, CH ₃ ,

R ^{2 is} H, CH ₃ ,

R ^{3 is} H, OH, OCH ₃ ,

R ^{4 is} H, O, OH, OCH ₃ , OC (O) CH ₃ and

R ^{5 is} H, O, OH, OCH ₃ , OC (O) CH ₃ ,

one of which contains enantiomers or diastereomers.

As particularly preferred compounds of the general formulas I to VI mentioned above, the following may be mentioned:

(2)

(10)

their enantiomers and their diastereomers.

The compounds of the general formulas I to VI were identified from the ginger fraction obtained according to the invention. To further characterize this ginger fraction and elucidate its ingredients, it was further suitably purified with the goal of isolating purified fractions of individual ingredients.

For this purpose, the ginger fraction obtained according to the invention was further purified by solid phase extraction on a C18 phase. The eluent of the solid phase extraction was brought to dryness and further analyzed by means of semipreparative high pressure liquid chromatography (HPLC). For this purpose, smaller aliquots of 5 to 10 mg were injected into the semipreparative HPLC system. The eluent of the HPLC column was then collected in 60 to 65 individual fractions and each of the resulting fractions was examined for their inhibitory effect on various P450 test reactions. The result of these investigations showed clearly demarcated zones (peaks) of higher inhibitory potency. •

To clarify the chemical structure of the constituents of the individual fractions, selected samples were further purified by repeated HPLC and concentrated and then analyzed by mass spectrometry and NMR spectroscopy.

The compounds (1) to (12) identified according to the invention are the typical ingredients of the ginger nonvolatile pungent fraction already sufficiently described in the literature. In addition to various modification products of the gingerole and its various homologues, a well-known main ingredient of ginger, the [8] -gingerol (12), and [6] -shogaol (9), the decomposition product of the quantitatively largest main ingredient [6] -gingerol , found. This was to confirm that the ginger fraction prepared by the described method is derived from the non-volatile high-energy fraction and the inhibition of the CYP enzymes is effected by ingredients of the gingerol type as well as its structural modifications and decomposition products.

A third aspect of the present invention is the use of the ginger fraction according to the invention and one or more of the compounds of the general formulas (I) to (VI) isolated therefrom for the production of a medicament for inhibiting cytochrome P450 enzymes, in particular cytochrome P450 3A4, 1A2, 2C19 and 2C9.

More preferably, the cytochromes P450 1A2, P450 2C19 and P450 2C9 are inhibited.

A fourth object of the present invention is the use of one or more compounds of the general formulas

wherein

n is the number 1, 2 or 3, R ^{1 is} H, CH ₃ ,

R ² H ₁ CH ₃ ,

R ^{3 is} H, OH, OCH ₃ ,

R ^{4 is} H, O, OH, OCH ₃ , OC (O) CH ₃ and

R ^{5 is} H, O, OH is ₁ OCH ₃ , OC (O) CH ₃ ,

their enantiomers or diastereomers for the preparation of a pharmaceutical composition for the inhibition of cytochrome P450 1A2, 2C9 and 2C19.

Particular preference may be given to the following compounds of general formulas I to IV:

their enantiomers and their diastereomers.

A fifth subject of the present invention is the use of the ginger fraction according to the invention and one or more of the compounds of the general formulas (I) to (VI) isolated therefrom in combination with a medicament for the production of a medicament for inhibiting human cytochrome P450 (CYP ) Enzymes, in particular cytochrome P450 3A4, 1A2, 2C19 and 2C9, for positively influencing the oral bioavailability and pharmacokinetics of active substances. More preferably, the cytochromes P450 1A2, 2C19 and 2C9 are inhibited.

Many drugs have low oral bioavailability due to so-called first-pass metabolism. This is the metabolic degradation of orally administered drugs in the small intestine or in the liver, even before they can reach the bloodstream to their active organ. In this case, the above-mentioned active substances, ie the pharmacologically active constituents of medicaments, may be selected from a group from medicines for influencing the cardiovascular system in the broadest sense, including substances that influence the composition of the blood (eg blood lipids); Medicaments with effect on the central nervous system; Medicines for the treatment of metabolic diseases (eg diabetes mellitus); Medicines for the treatment of inflammatory processes in the broadest sense; Medicines for influencing the immune system; Medicines for the treatment of infections by bacteria, unicellular organisms, multicellular parasites, viruses, fungi, or prions; Medicaments for the remedy or improvement of degenerative processes of various organs, in particular of the brain, and medicaments for the treatment of cancer.

The term "pharmaceuticals" means substances and preparations of substances which are intended, by application on or in the human or animal body, to heal, alleviate, prevent or treat diseases, ailments, physical injuries or pathological complaints detect;

2. to reveal the nature, condition or functions of the body or mental states;

3. replace active substances or body fluids produced by the human or animal body;

4. to ward off, eliminate or neutralize pathogens, parasites or foreign substances, or

5. to influence the nature, condition or functions of the body or mental states. ^'

Cytochrome P450 (CYP) enzymes are enzymes from the family of cytochrome P450 monooxygenases that are known to be involved in the metabolism of drugs. In particular, these are all P450 enzymes of the families CYP1A, CYP1B, CYP2A, CYP2B, CYP2C, CYP2D, CYP2E, CYP2F, CYP2J, CYP3A, CYP4A.

A sixth aspect of the present invention comprises a process for the preparation of a medicament for increasing the bioavailability of an oral administering a pharmaceutical compound comprising the oral administration of the pharmaceutical compound together with a ginger fraction according to any one of claims 8 to 10 to a person in need of such treatment, wherein the ginger fraction is administered in an amount necessary to increase the bioavailability of the pharmaceutical compound over the sole administration of the pharmaceutical compound.

The pharmaceutical compound is characterized by being metabolized by cytochrome P450 enzymes, preferably by P450 3A4, 1A2, 2C9 and 2C19.

A seventh subject of the present invention comprises a pharmaceutical formulation comprising the ginger fraction obtainable according to the invention or at least one compound of the general formulas I to VI, their enantiomers or their diastereomers, in addition to optionally one or more pharmaceutically acceptable excipients and / or diluents for improvement oral bioavailability and pharmacokinetics of drugs.

Preferably, such a pharmaceutical composition consists of two or more, optionally physically separate components and comprises:

(a) a first component consisting of the ginger fraction of the invention and one or more pharmaceutically acceptable diluents and / or carriers; and

(b) a second component comprising a pharmaceutical composition comprising a pharmaceutical compound metabolised by cytochrome P450 enzymes and one or more pharmaceutically acceptable diluents and / or carriers.

In a preferred pharmaceutical composition, the first component consists of at least one compound of the general formulas I to VI, their enantiomers or their diastereomers. In a further preferred pharmaceutical composition, the first component consists of at least one compound of the formulas (1) to (12), their enantiomers or their diastereomers.

The drug contained in the second component is preferably metabolized by the enzymes cytochrome P450 1A2, 3A4, 2C9 and 2C19.

As pharmaceutically acceptable carriers and / or diluents, corn starch, lactose, cane sugar, microcrystalline cellulose, magnesium stearate, polyvinyl pyrrolidone, citric acid, tartaric acid, water, water / ethanol, water / glycerol, water / sorbitol, water / polyethylene glycol, propylene glycol, cetylstearyl alcohol, Carboxymethylcellulose or fatty substances such as Hart¬ fat or their suitable mixtures are incorporated into common pharmaceutical preparations such as tablets, dragees, capsules, powders, suspensions, solutions, metered aerosols or suppositories.

Experimental part

10 g of an oleoresin (from Eramex Aromatics GmbH) are extracted by shaking three times with 50 ml of hexane each time and the supernatant (organic phase) is discarded. The

Residues are combined and shaken three times with 40 mL each 70 ⁰ C warm water. The supernatant is discarded again and the supernatant

Extract the residue three times with 40 mL of 70 ⁰ C warm methanol. The residue is discarded. The resulting supernatant is concentrated in a rotary evaporator and dissolved again in methanol.

Scheme 1 shows an overview of the extraction process of the ginger fraction of oleoresin according to the invention with removal of the essential oils.

Subsequently, the ginger fraction thus obtained was further purified by solid phase extraction on a C18 phase. The eluent of the solid phase extraction was brought to dryness and further investigated by means of semipreparative high-pressure liquid chromatography (HPLC). For this purpose, smaller aliquots of 5 to 10 mg were injected into the semipreparative HPLC system. The eluent of the HPLC column was then collected in 60 to 65 individual fractions and each of those obtained Fractions examined for their inhibitory effect on various P450 test reactions. The results of these investigations (Figures 1 to 4) showed clearly demarcated zones (peaks) of higher inhibitory potency.

Experiments with human liver microsomes

Test (A):

As a specific test reaction for CYP 3A4 erythromycin / V-demethylation is used.

The ginger fraction or different fractions of the extraction process are examined for their inhibition of CYP. To this end, 100 μg of human liver microsomes are incubated with 0.01 to 100 μg of ginger fraction and 7.34 μg (10 nmol) of erythromycin at pH 7.4 in the presence of NADPH. The inhibition of CYP activity is determined by comparison with control incubations without ginger extract (with equal solvent concentration).

Test B: As a specific test reaction for CYP 1A2, the phenacetin O-deethylation is used.

The ginger fraction or different fractions of the extraction process are examined for their inhibition of CYP. For this purpose, 125 mg of human liver microsomes are incubated with 0.01 to 100 mg of ginger fraction and 5 nmol of phenacetin at pH 7.4 in the presence of NADPH (1 mM) in a total volume of 250 μl. The inhibition of CYP activity is determined by comparison with control incubations without ginger extract (with equal solvent concentration).

Test C:

As a specific test reaction for CYP 2C19, S-mephenytoin 4 "hydroxylation is used. The ginger fraction or the various fractions of the extraction process are examined for their inhibition of CYP. For this purpose, 125 mg of human liver microsomes are incubated with 0.01 to 100 mg of ginger fraction and 12.5 nmol of S-mephenytoin at pH 7.4 in the presence of NADPH (1 mM) in a total volume of 250 μl. The inhibition of CYP activity is determined by comparison with control incubations without ginger extract (with the same solvent concentration).

Test D:

As a specific test reaction for CYP 2C9, tolbutamide hydroxylation is used.

The ginger fraction or different fractions of the extraction process are examined for their inhibition of CYP. For this purpose, 125 mg of human liver microsomes are incubated with 0.01 to 100 mg ginger fraction and 37.5 nmol tolbutamide at pH 7.4 in the presence of NADPH (1 mM) in a total volume of 250 μl. The inhibition of CYP activity is determined by comparison with control incubations without ginger extract (with the same solvent concentration).

Brief description of the pictures

Scheme 1 shows an overview of the extraction process of the ginger fraction according to the invention of an oleoresin with removal of the essential oils.

Figure 1 shows the HPLC separation of a ginger extract and the determination of the inhibitory potency against CYP3A4 for the eluted HPLC fractions (collection interval: 1 minute).

Figure 2 shows the HPLC separation of a ginger extract and the determination of the inhibitory potency against CYP1A2 for the eluted HPLC fractions (collection interval: 1 minute). Figure 3 shows the HPLC separation of a ginger extract and the determination of the inhibitory potency against CYP2C19 for the eluted HPLC fractions (collection interval: 1 minute).

Figure 4 shows the HPLC separation of a ginger extract and the determination of the inhibitory potency against CYP2C9 for the eluted HPLC fractions (collection interval: 1 minute).

Scheme 1: Overview of the extraction process of the ginger fraction of an oleoresin according to the invention with removal of the essential oils

Oleoresin (10 g)

Shaking with hexane (3x50 ml)

Backlog supernatant discarded

Shaking with water (IC ₅₀ : 22.6 μg / ml) (3 × 40 ml, 70 ^° C.)

Residue supernatant

(IC ₅₀ : 0.9 μg / ml) discarded (IC ₅₀ : 13.3 μg / ml)

Shake with methanol (3x 40 ml, 70 ⁰ C)

Residue rejected (no inhibition)

Claims

claims

A process for isolating a ginger fraction to remove the essential oil, comprising the steps

(a) extraction of an oleoresin with a nonpolar organic solvent;

(b) extraction of the combined residues from step (a) with warm water and rejection of the supernatant.

2. Process according to claim 1, characterized in that the combined residues obtained in step (b) are further purified by a process comprising the steps

(c) extraction with warm alcohol and

(d) concentrating the combined supernatants from step (c).

3. The method according to claim 1, characterized in that in step (a) as a non-polar organic solvent, a low-boiling alkane solvent, a petrochemical distillate, a fuel or other low-boiling, volatile and non-polar solvent is used.

4. The method according to claim 1, characterized in that in step (a) is used as a nonpolar organic solvent hexane.

5. The method according to claim 2, characterized in that in step (c) as alcohol methanol, ethanol, isopropanol, / 7-propanol, n-butanol or another positionally isomeric butanol, n-pentanol or a different position isomeric pentanol is used.

6. The method according to claim 2, characterized in that in step (c) is used as the alcohol methanol.

7. The method according to any one of claims 1 to 6, characterized in that the extractant used in steps (a), (b) and (c) is used in each case in amounts of 4 to 10 mL / g of the oleoresin used.

8. ginger fraction obtainable by a process according to any one of claims 1 to 7.

9. ginger fraction according to claim 8, characterized in that it comprises at least one compound of the general formulas

wherein n is the number 1, 2 or 3,

R ^{1 is} H, CH ₃ ,

R ² H ₁ CH ₃ ,

R ^{3 is} H, OH, OCH ₃ ,

R ^{4 is} H, O, OH, OCH ₃ , OC (O) CH ₃ and

R ^{5 is} H, O, OH, OCH ₃ , OC (O) CH ₃ ,

one of which contains enantiomers or diastereomers.

Ginger fraction according to claim 8 or 9, characterized in that it contains at least one of the following compounds:

their enantiomers and their diastereomers.

11. Use of the ginger fraction according to any one of claims 8 to 10 for the preparation of a pharmaceutical composition for the inhibition of cytochrome P450 enzymes.

12. Use of one or more compounds according to any one of claims 9 or 10, their enantiomers or their diastereomers for the preparation of a pharmaceutical composition for inhibiting human cytochrome P450 enzymes.

13. Use according to any one of claims 11 or 12, characterized in that the cytochromes P450 3A4, 1A2, 2C19 and 2C9 are inhibited.

14. Use according to one of claims 11 or 12, characterized in that the cytochromes P450 1A2, 2C9 and 2C19 are inhibited.

15. Use of one or more compounds of the general formulas

wherein n is the number 1, 2 or 3,

R ^{2 is} H, CH ₃ ,

R ^{3 is} H, OH, OCH ₃ ,

R ^{4 is} H, O, OH, OCH ₃ , OC (O) CH ₃ and

R ^{5 is} H, O, OH, OCH ₃ , OC (O) CH ₃ ,

16. Use according to claim 15, characterized in that the compounds of the general formulas I to IV are selected from

their enantiomers and their diastereomers.

17. Use of the ginger fraction according to any one of claims 8 to 10 in combination with a medicament for the preparation of a pharmaceutical composition for inhibiting human cytochrome P450 (CYP) enzymes.

18. Use of one or more compounds according to any one of claims 9 or 10, their enantiomers or their diastereomers in combination with a pharmaceutical composition for the preparation of a pharmaceutical composition for inhibiting human cytochrome P450 enzymes.

19. Use according to any one of claims 17 or 18, characterized gekenn¬ characterized in that the enzyme is cytochrome P450 3A4, 1 A2, 2C19 or 2C9.

20. Use according to one of claims 17 or 18, characterized in that the enzyme is cytochrome P450 1A2, 2C19 or 2C9.

21. A process for the preparation of a medicament for increasing the bioavailability of an orally administrable pharmaceutical compound comprising the oral

Application of the pharmaceutical compound together with a ginger fraction according to any one of claims 8 to 10 to a person in need of such treatment, wherein the ginger fraction is administered in an amount necessary to control the bioavailability of the pharmaceutical compound over the sole administration of the pharmaceutical compound.

22. The method according to claim 21, characterized in that the pharmaceutical compound is metabolized by cytochrome P450 3A4 enzymes.

23. The method according to claim 21, characterized in that the pharma¬ ceutical compound is metabolized by cytochrome P450 1A2 enzymes.

24. The method according to claim 21, characterized in that the pharmaceutical compound is metabolized by cytochrome P450 2C9 enzymes.

25. The method according to claim 21, characterized in that the pharma ceutical compound is metabolized by cytochrome P450 2C19 enzymes.

26. A pharmaceutical formulation comprising at least one compound of the general formulas I to VI, their enantiomers or their diastereomers, optionally together with one or more inert carriers and / or diluents.

27. Pharmaceutical formulation containing at least one compound of formulas (1) to (12), their enantiomers or their diastereomers, where appropriate, one or more inert carriers and / or diluents.

28. A pharmaceutical composition of two or more, optionally physically separated components, comprising: (a) a first component consisting of the ginger fraction according to any one of claims 8 to 10 and one or more pharmaceutically acceptable diluents and / or carriers; and

29. A pharmaceutical composition according to claim 28, characterized gekenn¬ characterized in that the first component contains at least one compound according to any one of claims 9 or 10, their enantiomers or their diastereomers.

30. A pharmaceutical composition according to any one of claims 28 or 29, characterized in that the pharmaceutical compound of the second component is metabolized by the enzymes cytochrome P450 1A2, 3A4, 2C9 or 2C19.