DRUG CONTAINING THYME AND PRIMROSE OR THYME AND IVY AS A PDE-4 INHIBITOR

Abstract

The invention relates to an extract from thyme (Thymus L.) in combination with primrose (Primula veris) or ivy (Hedera helix) for use as PDE-4 inhibitor in the prophylaxis and treatment of selected diseases and to a corresponding pharmaceutical and use thereof.

Claims

1-7. (canceled)

8. A method for prophylaxis and treatment of diseases of the asthmatic type which comprises administering the PDE-4 inhibitor as claimed in claim 8 to a person having an asthmatic type disease wherein PDE-4 is present in a galenic formulation which is selected from the group consisting of dragées, tablets, film-coated tablets, powder, capsules, drops, juices and syrups.

9. The method according to claim 8, wherein the disease is selected from the group consisting of atopic asthma, non-atopic asthma, allergic asthma, atopic bronchial IgE-mediated asthma, bronchial asthma, essential asthma, intrinsic asthma caused by pathophysiologic disturbances, extrinsic asthma caused by environmental factors, essential asthma of unknown or unapparent cause, bronchitic asthma, emphysematous asthma, exercise-induced asthma, allergen induced asthma, allergic asthma, cold air induced asthma, occupational asthma, infective asthma caused by bacterial, fungal, protozoal, or viral infection, non-allergic asthma, incipient asthma, and wheezy infant syndrome.

10. The method according to claim 8, wherein the disease is selected from the group consisting of bronchiolytis, chronic or acute bronchoconstriction, small airways obstruction, and emphysema, obstructive or inflammatory airways diseases, etiology, and pathogenesis.

11. The method according to claim 8, wherein the disease is selected from the group consisting of chronic eosinophilic pneumonia, adult respiratory distress syndrome (ARDS), exacerbation of airways hyper-reactivity consequent to other drug therapy and airways disease that is associated with pulmonary hypertension.

12. The method according to claim 8, wherein the disease is selected from the group consisting of diseases of acute laryngotracheal bronchitis, arachidic bronchitis, catarrhal. bronchitis, croupus bronchitis, dry bronchitis, infectious asthmatic bronchitis, productive bronchitis, staphylococcus or streptococcal bronchitis and vesicular bronchitis, acute lung injury, emphysema, post-traumatic pulmonary insufficiency postinfectious cough; etiology, and pathogenesis.

13. The method according to claim 8, wherein the disease is selected from the group consisting of cylindric bronchiectasis, sacculated bronchiectasis, fusiform bronchiectasis, capillary bronchiectasis, cystic bronchiectasis, dry bronchiectasis and follicular bronchiectasis.

14. The method according to claim 8, wherein the disease is selected from the group consisting of systemic lupus erythematosus, polyarteritis nodosa, Goodpasture's syndrome, scleroderma, wegner's granulomatosis, sjögren's syndrome, hamman-rich syndrome, sarcoidosis, dermatitis, dermatitis herpetiformis duhring, atopic dermatitis, psoriasis, and dermatomyositis.

15. The method according to claim 8, wherein said pharmaceutical formulation further comprises a carrier substance.

16. The method according to claim 15, wherein the disease is a postinfectious cough.

17. The method according to claim 16, wherein said pharmaceutical formulation further comprises an active substance, a filler and an extender.

18. The method according to claim 16, wherein the carrier substance is polyethylene glycol, cocoa fat, C.sub.14 alcohol with C.sub.16 fatty acid or mixtures thereof.

19. The method according to claim 16, wherein the carrier substance is animal and plant fat, wax, paraffin, starch, astragalus cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talcum, zinc oxide or mixtures thereof.

20. The method according to claim 16, wherein the carrier substance is lactose, talcum, silica, aluminium hydroxide, calcium silicate and polyamide powder or mixtures thereof.

21. The method according to claim 16, wherein the carrier substance is a solvent, solubilising agent, emulsifier or mixtures thereof.

22. The method according to claim 16, wherein the carrier substance is water, ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, cotton seed oil, groundnut oil, corn oil, olive oil, castor oil, and sesame oil, glycerol, glycerol formal, tetrahydrofurfuryl alcohol, polyethylene glycol, fatty acid esters of sorbitan or mixtures thereof.

23. The method according to claim 16, wherein the carrier is liquid diluent, suspension agent, microcrystalline cellulose, aluminium metahydroxide, bentonite, agar-agar, astragalus or mixtures thereof.

24. The method according to claim 16, wherein the carrier is ethyl alcohol, propylene glycol, ethoxylated isostearyl alcohols, polyoxyethylene sorbitan ester, sorbitan ester, microcrystalline cellulose, aluminium metahydroxide, bentonite, agar-agar, astragalus, or mixtures of thereof.

25. The method according to claim 8, wherein the disease is postinfectious cough and the galenic formulation is a syrup.

26. The method according to claim 8, wherein the individual takes, 1, 2, 3 or 4 doses a day of the PDE-4 inhibitor.

Description

A BRIEF DESCRIPTION OF THE FIGURES

[0026] FIG. 1 illustrates the inhibition of the phosphodiesterase-4 activity,

[0027] FIG. 2 illustrates the treatment with BRO-TP (supra) led from 100 μg/mL to a considerable rise in CBF compared with thyme or primrose alone.

[0028] FIG. 3 illustrates the samples for TNFalpha release in vitro (anti-inflammation).

[0029] FIG. 4 illustrates the Leukotriene formation and 5-LO activity.

[0030] In accordance with the invention it is also advantageous that thyme and ivy or thyme and primrose or a respective extract thereof allow an increase of the mucocillary clearance. Mucocillary clearance is a mechanism in which both foreign particles and mucus from bronchi and the windpipe are transported away upwardly. For this purpose, the epithelial cells are provided with cilia, which beat rhythmically and move the mucus along. A restricted beat frequency (CEF) leads to a weakening of the transporting away of the mucus and is compensated for by coughing. Driving the beat frequency can therefore remedy coughing and reduce the mucus-conveyed obstruction of the airways. Surprisingly, thyme and ivy or thyme and primrose or a respective extract thereof demonstrate an advantageous induction of the ciliary beat frequency, such that the curative and palliative effect of the diseases linked by PDE-4 inhibitors is positively assisted.

[0031] In addition, thyme and ivy or thyme and primrose or a respective extract thereof demonstrate an advantageous secondary effect of protection against inflammation, as evidenced in Example 4 and FIG. 4, which is provided in addition to PDE-4 inhibition and substantiates a supporting rationale for the aforementioned indications according to the invention.

[0032] The specified plant (drugs) according to the invention for the pharmaceutical according to the invention can be obtained, as is conventional for the respective plant drug, from preferred plant parts, such as the leaf, root, etc., as described correspondingly (for example see Ph.Eur., European Pharmacopoeia). In particular, aqueous and/or ethanolic extracts can be used or dry extracts obtainable therefrom. Reference is made to the technical teaching forming the subject matter of EP 1368605B1 and EP 0753306B1 with regard to the production of the extracts according to the invention.

[0033] The pharmaceuticals of the present invention can be produced advantageously and in the manner known per se and can be used in the form of a galenic formulation corresponding to their application.

[0034] The galenic formulations of the pharmaceuticals according to the invention are characterised in that they include oral formulations such as dragées, tablets, film-coated tablets, powder, capsules or liquid dilutions, in particular drops, juices or syrups.

[0035] In the case of topical application or use, sprays, ointments, emulsions, powders, grains, liquid or solid preparations for inhalation, compresses, tamponades, tonsil brush solutions or gargling solutions are suitable in particular.

[0036] The invention also relates to a pharmaceutical formulation containing a pharmaceutical according to the invention. The pharmaceuticals according to the invention can be prepared in the form of pharmaceutical preparations or compositions in metred units. This means that the preparation or composition is present in the form of individual parts, for example tablets, dragées, capsules, pills, suppositories and ampoules, of which the active substance content corresponds to a fraction or a multiple of an individual dose. The metred units may contain, for example, 1, 2, 3 or 4 individual doses or ½, ⅓ or ¼ of an individual dose. An individual dose preferably contains the quantity of active substance which is administered with an application and which usually corresponds to an entire daily dose, half a daily dose, a third of a daily dose or a quarter of a daily dose.

[0037] The term non-toxic, inert pharmaceutically suitable carrier substances is to be understood to mean solid, semi-solid or liquid diluents, fillers and formulation aids of any type.

[0038] The term pharmaceutical also includes agents of identical suitability, such as food supplements or even foodstuffs, provided the advantageous effect according to the invention of the agent is implemented.

[0039] Preferred pharmaceutical formulations are tablets, dragées, capsules, pills, granulates, suppositories, solutions, juice, suspensions and emulsions, pastes, ointments, gels, creams, lotions, dusts and (nasal) sprays. Tablets, dragées, capsules, pills and granulates may contain the active substance(s) in addition to the conventional carrier materials, such as a) fillers and extenders, for example starches, lactose, sucrose, glucose, mannitol and silica, b) binders, for example carboxymethyl cellulose, alginates, gelatins, polyvinyl pyrrolidone, c) humectants, for example glycerol, d) exploders, for example agar-agar, calcium carbonate and sodium carbonate, e) solution retarders, for example paraffin, and f) resorption accelerators, for example quaternary ammonium compounds, g) wetting agents, for example cetyl alcohol, glycerol monostearate, h) adsorption agents, for example kaolin and bentonite, and i) lubricants, for example talcum, calcium stearate and magnesium stearate, and solid polyethylene glycols, or mixtures of the substances specified under a) to i).

[0040] The tablets, dragées, capsules, pills, and granulates may be provided with the usual coatings and casings, which may or may not contain opacifying agents, and may also be composed such that they only or preferably only deliver the active substances in a certain part of the intestinal tract in a delayed manner as necessary, wherein polymer substances and wax for example can be used as embedding compounds. The active substance(s) may also be present in microencapsmlated form, where necessary with one or more of the above-mentioned carrier substances.

[0041] Suppositories, besides the active substance(s), may also contain conventional water-soluble or water-insoluble carrier substances, for example polyethylene glycols, fats, for example cocoa fat and higher esters (for example C14 alcohol with C16 fatty acid) or mixtures of these substances).

[0042] Ointments, pastes, creams and gels, besides the active substance(s), may also contain the conventional carrier substances, for example animal and plant fats, waxes, paraffins, starch, astragalus cellulose derivatives, polyethylene glycols, silicones, bentonites, silica, talcum and zinc oxide or mixtures of these substances.

[0043] Powders and sprays, besides the active substance(s), may also contain the conventional carrier substances, for example lactose, talcum, silica, aluminium hydroxide, calcium silicate and polyamide powder or mixtures of these substances. Sprays may additionally contain the conventional propellants. Solutions and emulsions, besides the active substance(s), may also contain the conventional carrier substances, such as solvents, solubilising agents, and emulsifiers, for example water, ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oils, in particular cotton seed oil, groundnut oil, corn oil, olive oil, castor oil, and sesame oil, glycerol, glycerol formal, tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid esters of sorbitan, or mixtures of these substances.

[0044] Suspensions, besides the active substance(s), may also contain the conventional carrier substances, such as liquid diluents, for example water, ethyl alcohol, propylene glycol, suspension agents, for example ethoxylated isostearyl alcohols, polyoxyethylene sorbitan esters and sorbitan esters, microcrystalline cellulose, aluminium meta hydroxide, bentonite, agar agar, and astragalus, or mixtures of these substances. The specified formulation forms may also contain dyes, preserving agents, and smell- or taste-enhancing additives, for example peppermint oil and eucalyptus oil and sweeteners, for example saccharin.

[0045] Further advantages and features of the present invention will emerge on the basis of the description of exemplary embodiments. The following examples and figures serve to explain the invention, without limiting the invention to these examples.

[0046] 1. Inhibition of the Phosphodiesterase-4 Activity

[0047] In an in vitro standard experiment it could be shown that the mixture of thyme and primrose dry extract (“BRO-TP”) is able to inhibit the activity of the catalytic domain of the PDE4 enzyme in a concentration-dependent manner. IC50 was 21-26 μg/mL. The individual extracts likewise inhibited the activity of the catalytic domain (thyme: approximately 20 μg/mL; primrose: 10-12 μg/mL). In the further experiments the specific effect on the PDE4 subtypes PDE4B and PDE4D was examined. Here too, inhibitory activity could be demonstrated for all extracts: in the case of PDE4B2, the IC50 values of BRO-TP, thyme and primrose were 26, 22 and 51 μg/mL, and for PDE4D3 were 17, 11 and 28 μg/ml, (see FIG. 1).

[0048] Methodology: The catalytic domain (PDE4Cat) and the enzyme subtypes PDE4B2 and -D3 were incubated with 6 concentrations of BRO-TP (3.07-300 μg/mL), and the catalytic activity was determined in a radiometric assay. The active substance rolipram (supra) was used as positive control.

[0049] 2. Induction of Ciliary Beat Frequency (CBF)

[0050] In an ex vivo experiment thin lung slices (Precision-Cut Lung Slices—PCLS) were treated with the thyme/primrose extract mixture, and the individual extracts and the beat frequency of the epithelial cilia were analysed. The treatment with BRO-TP (supra) led from 100 μg/ml, to a considerable rise in CBF compared with thyme or primrose alone (see FIG. 2).

[0051] Methodology: PCLS 300 μm thick were obtained from guinea pig lungs and transferred into 24-well plates with medium. Rising concentrations of the extracts were added to the medium (10 μg/mL, 32 μg/mL, 100 μg/mL, 320 μg/mL, and 1000 μg/mL) and the beat frequency at the ciliary edge of the airways was analysed by video microscope. By way of comparison, the influence of the vehicle (0.005-0.5% ethanol) was determined. The maximum frequency rise was determined by addition of methacholine (10 μM).

[0052] It could be shown that the anti-inflammatory effect of the thyme/primrose dry extract mixture is conveyed inter alia via an inhibition of the PDE4 enzyme activity. In addition, an ex vivo cilial beat-stimulating effect is produced by the thyme extract and can be caused by the extract mixture. The results were ascertained in models that are relevant for the pathogenesis and therapy of the claimed indications and therefore provide sufficient proof that in particular the combination of thyme and primrose (dry) extract, as contained in Bronchipret®, is also effective in the treatment of patients.

[0053] 3. BRO-TP—TNFalpha Release In Vitro (Anti-Inflammation)

[0054] Model: TNFalpha (tumour necrosis factor alpha) release from LPS (lipopolysaccharide)-stimulated immune cells

[0055] Protocol—LPS (dexamethasone): Human PBMC in 96-well plates in culture medium (RPMI 1640, 10% FBS, 1% Pen/Strep, 2 mM L-alanyl-L-glutamine), cell density: 5E.sup.4 cells/well (cytokine plate), 2E.sup.5 cells/well (alamarBlue plate). Incubation of the cells for 1 h at 37° C. in 5% CO.sub.2, addition of the components and dexamethasone (vehicle control=0.15% EtOH), incubation of the plates for 1 h at 37° C. in 5% CO.sub.2, subsequent addition of LPS and incubation of the plates for 24 h at 37° C. in 5% CO.sub.2, centrifugation (1,000 rpm, 10 min.), collection of the supernatants and freezing at −80° C.

[0056] The samples were assessed and the data is presented in FIG. 3.

[0057] Summary; BRO-TP inhibits the release of the pro-inflammatory cytokine TNFa from UPS-stimulated human immune cells

[0058] 4. Leukotriene Formation and 5-LO Activity

[0059] Freshly isolated human neutrophils (5×10.sup.6 cells/ml for LTB.sub.4 formation) or monocytes (1×10.sup.6 cells/ml for cys-LT formation) were incubated in PGC buffer for 10 min at 37° C. (with vehicle (0.025% EtOH)), THY/PRI, THY, PHI (0.4-100 μg/mL in each case) or zileuton (3 μM) and then stimulated for 10 min at 37° C. with ionophore A23187 (2.5 μM). The reaction was stopped with ice, and suitable reagents for LTB.sub.4 analysis or for cys-LT analysis were added. LTB.sub.4 was analysed using HPLC. Cys-LTs was analysed using ELISA. The results are presented in FIG. 4.

[0060] 5-LO was expressed in transformed E. coli B121 and purified using ATP affinity chromatography. In order to determine the 5-LO activity, enzymes were added in a reaction mixture to 5-LO. Samples were incubated for 10 min at 4° C. (with vehicle (0.25% Et0H)), and THY/PRI, THY, PRI (0.4-100 μg/ml in each ease), or zileuton (3 μM), preheated at 37° C., and CaCl.sub.2 and arachidonic acid (20 μM) were added. The reaction was stopped after 10 min at 37° C., and prostaglandin B1 (internal standard) was added. Obtained metabolites were analysed using HPLC, more specifically 5-LO products inc. LTB.sub.4 all-trans isomers and 5-H(P)ETE. The results are presented in FIG. 4.

[0061] Summary: BRO-TP demonstrates improved anti-inflammatory effects (5-LO) compared with thyme and primrose alone.