Aug 2005PROSPAN ® 1 A Herbal Preparation with a Proven Mode of Action

Aug 2005PROSPAN ® 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

Aug 2005PROSPAN ® 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 productsHerbal products

Aug 2005PROSPAN ® 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 productsHerbal products

Aug 2005PROSPAN ® 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 productsHerbal products

Aug 2005PROSPAN ® 6 Differences in medicinal products Herbal – Herbal Herbal extracts of the same plant may be different in efficacy and tolerability !

Aug 2005PROSPAN ® 7 Differences in medicinal products Herbal – Herbal Herbal extracts are characterized by:  Kind of extract –Fluid extract –Spissum extract –Dried extract

Aug 2005PROSPAN ® 8 Differences in medicinal products Herbal – Herbal  Extractive agent –Influences the kind and amount of extracted substances (lipohilic or hydrophilic) Herbal extracts are characterized by:

Aug 2005PROSPAN ® 9 Differences in medicinal products Herbal – Herbal  Drug-Extract-Ratio (DER) –How many drug is used to get 1 g of extract? More or less concentrated! Herbal extracts are characterized by:

Aug 2005PROSPAN ® 10 Differences in medicinal products Herbal – Herbal  Composition of the extract itself –respective to quality and quantity of all contained substances Herbal extracts are characterized by:

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

Aug 2005PROSPAN ® 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

Aug 2005PROSPAN ® 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!

Aug 2005PROSPAN ® 14 PROSPAN ® Dried extract of Ivy leaves (DER : 1) Efficacy and safety for this extract has been proved in various clinical studies secretolyticbroncholyticcough relieving

Aug 2005PROSPAN ® 15 PROSPAN ® Objective (Lung function) - vital capacity (VC), - forced vital capacity (FVC), - forced expiratory volume/sec. (FEV 1 ), - 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 Documented improvements

Aug 2005PROSPAN ® 16 „Ivy: mode of action evidenced by cell biological investigations“ In cooperation with Prof. Dr. Hanns Häberlein Physiological Chemistry University of Bonn

Aug 2005PROSPAN ® 17 Main constituents of Hedera helix L.  -hederin: R = H hederacoside C: R = 1(β-D-glucose)6 – 1(β-D-glucose)4 – 1(α-L-rhamnose) Triterpensaponins

Mode of action of Ivy indirect increase of ß 2 -adrenergic effects ß 2 -adrenergic receptor key role lamellar bodies cAMP surfactant PKA lung cell cAMP SR [Ca 2+ i  ] Ca 2+ channel unstriated muscle cell

Endocytosis Regulation of β 2 -adrenergic receptor density Degradation Recycling Ligand (L) Accumulation of RL - complexes in „coated pits“ β 2 -adrenergic receptor (R) RL-complex in „lipid rafts“ Degradation early endosome

endocytosis How to increase ß 2 -adrenergic effects on living cells? degradation recycling ligand (L) accumulation of receptor-ligand- complexes (coated pit) clathrin early endosome ß 2 -adrenergic receptor (R) RL-complex in lipid rafts α-hederin

Aug 2005PROSPAN ® 21 Fluorescence Correlation Spectroscopy

Aug 2005PROSPAN ® 22 FCS: Free ligand Cell membrane Detection volume Ligand Diffusion time of free ligand 45 µs Alexa-NA (ß 2 -adrenergic agonist)

Aug 2005PROSPAN ® 23 FCS: Ligand-Receptor-Complex Diffusion time of ligand-receptor-complex 3.3 ms Cell membrane Detection volumeReceptor-ligand complex

Aug 2005PROSPAN ® 24 Diffusion time of accumulated ligand-receptor-complex 95 ms Cell membrane Detection volume FCS: Accumulated Ligand-Receptor-Complex Accumulated receptor-ligand complex

Aug 2005PROSPAN ® 25 Free ligand Receptor-ligand-complex Accumulated complex

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

Aug 2005PROSPAN ® 27  -hederin influences regulatory processes of ß 2 -adrenergic receptors:  -hederin inhibits redistribution as well as internalisation of even redistributed ß 2 - adrenergic receptors after ligand binding. Prospan: mode of action

Aug 2005PROSPAN ® 28 An increased ß 2 -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). Ivy: Mode of action – consequences I lamellar bodies cAMP surfactant PKA ß 2 -adrenergic receptor

Aug 2005PROSPAN ® 29 An increased ß 2 -adrenergic receptor density and an increased signal transduction lead to an increased production of cAMP:  decrease in intracellular [Ca 2+ i ] with subseeding bronchial muscle relaxation (formation of less active myosin kinase via phosphorylation by phosphokinase A). cAMP SR [Ca 2+ i  ] Ca 2+ channel ß 2 -adrenergic receptor Ivy: Mode of action – consequences II

Aug 2005PROSPAN ® 30 Ivy - Resorption In vitro (CaCo-2-cells) Transport of Hederacosid C Transport of alpha-hederin Time (min)

Aug 2005PROSPAN ® 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

Aug 2005PROSPAN ® 32 Ivy- mode of action  -hederin reduction of mucus viscosity dilatation of bronchial musculature increased β 2 –adrenergic stimulation lung epitheliumbronchial muscle surfactant- production Ca ++ (intracellular) secretolytic broncholytic Expectorant

Aug 2005PROSPAN ® 33 Ivy: Effect on ß 2 -receptors in general In theory  -hederin supports indirectly the stimulaton of all ß 2 -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)

Thank you very much for your kind attention!