PROSPAN ®

1. PROSPAN® A Herbal Preparation with a Proven Mode of Action

2. Chemical products First chemical products derived from herbal pattern Herbal products Long-time experience in phytotherapy Application of plants or parts of plants is historically the basis for any therapy Differences in medicinal products Chemical – Herbal

3. Defined substance(s) as active ingredient(s) This active ingredient is chemically synthetizised and exactly characterized Whole plant extract as active ingredient These extracts are complex multisubstance mixtures Differences in medicinal products Chemical – Herbal Chemical products Herbal products

4. Defined doses of the active ingredient must have equivalent efficacy in every final product Pharmacokinetic studies are easy to perform due to monosubstance character Every substance of the extract may contribute to the efficacy (and tolerability) of the extract Pharmacokinetic studies are nearly impossible to perform due to multisubstance character Differences in medicinal products Chemical – Herbal Chemical products Herbal products

5. Products with the same active ingredient must guarantee the same efficacy proven by studies respective to bioavailability or bioequivalence Products with an extract of the same plant from different manufacturers may differ in efficacy and tolerability Differences in medicinal products Chemical – Herbal Chemical products Herbal products

6. Differences in medicinal products Herbal – Herbal Herbal extracts of the same plant may be different in efficacy and tolerability !

7. Differences in medicinal products Herbal – Herbal Herbal extracts are characterized by: • Kind of extract • Fluid extract • Spissum extract • Dried extract

8. Differences in medicinal products Herbal – Herbal Herbal extracts are characterized by: • Extractive agent • Influences the kind and amount of extracted substances (lipohilic or hydrophilic)

9. Differences in medicinal products Herbal – Herbal Herbal extracts are characterized by: • Drug-Extract-Ratio (DER) • How many drug is used to get 1 g of extract? More or less concentrated!

10. Differences in medicinal products Herbal – Herbal Herbal extracts are characterized by: • Composition of the extract itself • respective to quality and quantity of all contained substances

11. Dependency of the composition of a herbal extract from manufacturing and quality parameters drug extracting agent content of active substance specific extracting agent content of water concentration cutting size amount portion of powder flow rate homogeneity herbal extract extraction time filling quantity extraction pressure extraction temperature filling height / density method of extraction batch size static pressure manufact. process facility

12. Differences in medicinal products Herbal – Herbal • Efficacy and safety of a herbal extract depends on its very special quality! • Extracts from the same part of the same plant can show different clinical properties

13. Differences in medicinal products Herbal – Herbal Results of clinical studies with an extract of manufacturer A are not automatically valid for the extract of manufacturer B The preparation of an extract and the production process for the medicinal product is based on a very special knowledge in each company!

14. PROSPAN® Dried extract of Ivy leaves (DER 5-7.5 : 1) Efficacy and safety for this extract has been proved in various clinical studies secretolytic broncholytic cough relieving

15. Objective (Lung function) vital capacity (VC), forced vital capacity (FVC), - forced expiratory volume/sec. (FEV1), - intrathoracic gas volume (ITGV), - residual volume (RV), airway resistance (RAW, obstruction marker) - peak expiratory flow (PEF) Subjective - coughing frequency - coughing intensity - painful coughing - sputum production - expectoration - dyspnoea - general well-being PROSPAN® Documented improvements

16. „Ivy: mode of action evidenced by cell biological investigations“ In cooperation with Prof. Dr. Hanns Häberlein Physiological Chemistry University of Bonn

17. Main constituents of Hedera helix L. Triterpensaponins hederacoside C: R = 1(β-D-glucose)6 – 1(β-D-glucose)4 – 1(α-L-rhamnose) a-hederin: R = H

18. key role lamellar bodies Ca2+ channel surfactant cAMP cAMP PKA SR [Ca2+i] lung cell unstriated muscle cell Mode of action of Ivy indirect increase of ß2-adrenergic effects ß2-adrenergic receptor

19. Endocytosis Regulation of β2-adrenergic receptor density Accumulation of RL - complexes in „coated pits“ Ligand (L) RL-complex in „lipid rafts“ β2-adrenergic receptor (R) Recycling early endosome Degradation Degradation

20. How to increase ß2-adrenergic effects on living cells? accumulation of receptor-ligand-complexes (coated pit) ligand (L) RL-complex in lipid rafts ß2-adrenergic receptor (R) clathrin recycling endocytosis early endosome degradation degradation α-hederin

21. Fluorescence Correlation Spectroscopy

22. Detection volume Ligand Ligand Cell membrane Alexa-NA (ß2-adrenergic agonist) FCS: Free ligand Diffusion time of freeligand 45 µs

23. Detection volume Receptor-ligandcomplex Cell membrane FCS: Ligand-Receptor-Complex Diffusion time ofligand-receptor-complex3.3 ms

24. Detection volume Accumulated receptor-ligandcomplex Cell membrane FCS: Accumulated Ligand-Receptor-Complex Diffusion time ofaccumulated ligand-receptor-complex95 ms

25. Receptor-ligand-complex Free ligand Accumulated complex

26. control, untreated pretreatment with 1µM -hederin for 24 h, then 10 µM terbutaline 20 min 10µM terbutaline 20 min Inhibition of internalization of 2-adrenergic receptors in pulmonary epithelial cells (A549) by -hederin.

27. Prospan: mode of action -hederin influences regulatory processes of ß2-adrenergic receptors: -hederin inhibits redistributionas well asinternalisation of even redistributed ß2-adrenergic receptors after ligand binding.

28. lamellar bodies surfactant ß2-adrenergic receptor cAMP PKA Ivy: Mode of action – consequences I • An increased ß2-adrenergic receptor density and an increased • signal transduction lead to an increased production of cAMP: • increased exocytosis of surfactant in pulmonary epithelial cells (alveolar type II cells) (secretolytic effect,decrease in mucus viscosity, decrease in coughing intensity and frequency).

29. Ca2+ channel cAMP ß2-adrenergic receptor [Ca2+i] SR Ivy: Mode of action – consequences II • An increased ß2-adrenergic receptor density and an increased • signal transduction lead to an increased production of cAMP: • decrease in intracellular [Ca2+i] with subseeding bronchial muscle relaxation (formation of less active myosin kinase via phosphorylation by phosphokinase A).

30. Ivy - Resorption In vitro (CaCo-2-cells) Transport of Hederacosid C Transport of alpha-hederin Time (min) Time (min)

31. Ivy - Resorption In vivo – first results • alpha-hederin • discovered in blood of treated animals and humans • the amount of hederacosid C given in an extract seems to support the concentration of alpha-hederin in blood (prodrug??) • Actually: Ongoing works on the sensitivity of analytical methods for further clarification

32. -hederin increased β2 –adrenergic stimulation lung epithelium bronchial muscle surfactant- production Ca++ (intracellular) secretolytic broncholytic dilatation of bronchial musculature reduction of mucus viscosity Ivy- mode of action Expectorant

33. Ivy: Effect on ß2-receptors in general In theory -hederin supports indirectly the stimulaton of allß2-receptors but given by the smooth and indirect effect, a result will only be seen in those organs with a pathological condition (e.g. ivy will have no bronchiolytic effect in case of „normal“ bronchial muscles)

34. Thank you very much for your kind attention!