DPhG Weihrauch-Symposiumanlässlich des 75. Geburtstages

Prof. Dr. Dr. h.c.mult. Hildebert Wagner – Center of Pharma Research – Department of Pharmacy – University of Munich Folie 1

Prof. Dr. Dr. h.c.mult. Hildebert Wagner

Center of Pharma Research – Department of Pharmacy – University of Munich

e-mail: h.wagner@cup.uni-muenchen.de

von Prof. Dr. H. P. T. Ammon

5. April 2008

Boswellia and the Complement System:A multiherbal drug for multitarget therapy

Prof. Dr. Dr. h.c.mult. Hildebert Wagner – Center of Pharma Research – Department of Pharmacy – University of Munich Folie 2

Prof. Dr. Dr. h.c.mult. Hildebert Wagner – Center of Pharma Research – Department of Pharmacy – University of Munich Folie 3

Komplement-System

Bindeglied zwischen humoralen und zellulären Reaktionsketten in angeborenen und adaptiven Immunantworten

Klassischer Weg Alternativer WegInduktion über Binding an Antikörper auf

pathogenen ZellenInduktion durch Antikörper unabhängige

direkte Bindung an pathogene Zellen

Prof. Dr. Dr. h.c.mult. Hildebert Wagner – Center of Pharma Research – Department of Pharmacy – University of Munich Folie 4

Funktionen und biologische Wirkungen von Komplementfragmenten

n Eliminierung von Pathogenen (Bakterien, Viren, Tumorzellen) durch Opsonierung, Phagozytose und Lyse.

Prof. Dr. Dr. h.c.mult. Hildebert Wagner – Center of Pharma Research – Department of Pharmacy – University of Munich Folie 5

n Inhibierung einer Anaphylotoxin-Bildung (C5a, C3a, C4a-Komplementfaktoren) durch Hemmung der Komplementaktivierung (z.B. durch Boswelliasäuren)

n Stimulierung der Chemotaxis von Granulozyten und Makrophagen Eliminierung von Antigen-Antikörper-Komplexen ("Immunkomplexen")

Boswelliasäuren

H

H CH3

CH3

R2

CH3

CH3H3C

Prof. Dr. Dr. h.c.mult. Hildebert Wagner – Center of Pharma Research – Department of Pharmacy – University of Munich Folie 6

ββββ-Boswelliasäure: R1, R2 : H

Acetyl-ββββ-Boswelliasäure: R1 : – C – CH3, R2 : H

11-Keto-ββββ-Boswelliasäure: R1 : H, R2 : = O

Acetyl-11-Keto-ββββ-Boswelliasäure: R1 : – C – CH3, R2 : = O

O

=

O

=

HCOOH

H3C

R1O

Inhibition of the classical pathway (CP) and the alternative pathway (AP) of the complement system by

triterpenes: reduction of hemolysis (%)

Glycyrrhetic acid Oleanolic acid Ursolic acidCucurbitacin I

Concentration of test compounds

Prof. Dr. Dr. h.c.mult. Hildebert Wagner – Center of Pharma Research – Department of Pharmacy – University of Munich Folie 7

ββββ-Boswellic acid

Crataegolic acidQuillaic acid

50 – 60% Harz5 – 9% Ätherischöl

Inhaltsstoffe von Boswellia carteriGummmiresinol / Olibanum

n 1-Octylacetat

n 1-Octanol

n (Iso)Cembren

n (Iso)-Incensol

n ββββ-Boswelliasäure

n Acetyl- ββββ-Boswelliasäure

n 11-Keto- ββββ-Boswelliasäure

n Acetyl-11-Keto- ββββ-

Prof. Dr. Dr. h.c.mult. Hildebert Wagner – Center of Pharma Research – Department of Pharmacy – University of Munich Folie 8

Polysaccharide

n (Iso)-Incensol

n αααα, ββββ-Pinen

n Cadinen

n Campher

n αααα-Phellandren

n Borneol

n Verbenon

n (+)Carvon u.a.)

n Acetyl-11-Keto- ββββ-Boswelliasäure

n Tirucallsäure

n Homo-Heteroglucane

Fact

Several standardized phytoextract preparations (multidrug mixtures) show

Prof. Dr. Dr. h.c.mult. Hildebert Wagner – Center of Pharma Research – Department of Pharmacy – University of Munich Folie 9

at appropriate indications and dosages therapeutic equivalence with chemotherapeutics but with

less or lacking adverse effects in comparison to chemotherapeutics

What could be the causes ofthis therapy equivalence?

n Synergistic and multitarget actions of bioactive constituents of the extracts

n Enhancement of the solubility / resorption rate and thereby the bioavailability of bioactive compounds by concomitant

Prof. Dr. Dr. h.c.mult. Hildebert Wagner – Center of Pharma Research – Department of Pharmacy – University of Munich Folie 10

n Elimination or "neutralization" of adverse acting substances of extracts

n Interaction with resistance mechanisms of pathogenic microorganisms

the bioavailability of bioactive compounds by concomitant constituents of the extracts

Drug-synergism of phytopharmaceuticals

plant extract

Prof. Dr. Dr. h.c.mult. Hildebert Wagner – Center of Pharma Research – Department of Pharmacy – University of Munich Folie 11

overadditive, potentiated pharmacological effects

additive, agonistic pharmacological effects

different targets(multitargeting)

one target

n A total effect of a combination is greater than expected from the sum of the individual agents

E (da,db) > E (da) + E (db)

Definition of synergy according to Berenbaum (Pharmacol. Rev. 41:93-141,1989)

Prof. Dr. Dr. h.c.mult. Hildebert Wagner – Center of Pharma Research – Department of Pharmacy – University of Munich Folie 12

E = observed effect da and db = doses of agents a and b

n The effect of a combination is greater than that of each of the individual agents

E (da,db) > E (da) and E (da,db) > E (db)

Ginkgolide A [µM]

Ginkgolide B [µM]

14

12

10

8

6

4

2

O O

OH

O

O

O

OO

HO

HO

OH

OH

H

O O

OH

O

O

O

OO

HO

HO

OH

H

H

Isobol-curve for 50% inhibition of a Ginkgolide AB-combination*

Prof. Dr. Dr. h.c.mult. Hildebert Wagner – Center of Pharma Research – Department of Pharmacy – University of Munich Folie 13

IC50 – values for various dose-combinations of PAF-induced thrombocyte aggregation*

GA : GB

3 : 12 : 11 : 11 : 21 : 31 : 10

IC50 [µg/ml]

2.402.201.801.551.401.30

Ginkgolide A [µM]

4.413.602.211.270.880.29

Ginkgolide B [µM]

1.421.722.122.432.572.79

Ginkgolide B [µM]0

0.5 1 1.5 2 2.5 3 3.5

O H

Ginkgo biloba: Ginkgolide mixtures/ Chung et al. (1987)Ginkgo extract

Piper methysticum: Kava lactones/mixtures Singh and Blumental (1997)of Kava lactones and extract fractions

Glycyrrhiza glabra: Licorice extract potentiates Cantelli-Forti et al. (1994)

In vitro and in vivo pharmacological evidences for synergy effects

(according to Williamson, Phytomedicine 8(5):401-409 (2001)

Prof. Dr. Dr. h.c.mult. Hildebert Wagner – Center of Pharma Research – Department of Pharmacy – University of Munich Folie 14

Glycyrrhiza glabra: Licorice extract potentiates Cantelli-Forti et al. (1994)other substances and acts Kimura et al. (1992)as detoxifier Miaorong and Jing (1996)

Cannabis sativa: Cannabis extract / THC Zuardi et al. (1982)Baker et al. (2000)

Valeriana offic.: Valeriana extract/ Hölzl (1997)individual constituents

Zingiber offic.: Zingiber extract/ Beckstrom-Sternberg andmixture of volatile terpenoids Duke (1994)and mixtures

Kava-kava + Passiflora incarn. Capasso and Sorrentino (2005)

Presynaptic neuron

H+

Na++ ?

COMTMAO

?

β-adr.5-HT1A/2ADA2,3,4 NMDA

Scheme of proposed biological targets of Hypericum perforatum according to in vitro

studies

+_

_

_

HyperforinHypericum

FlavonoidsXanthones

Hypericum

Prof. Dr. Dr. h.c.mult. Hildebert Wagner – Center of Pharma Research – Department of Pharmacy – University of Munich Folie 15

_

Postsynaptic neuron

Hypothalamus

Cortisol

IL-6

Adrenal cortexPituitaryACTH

Prolactin

CRF

CRF

TRH

DA2,3,4 NMDA

Opioid

GABAA

BenzoSigma

5-HT6,7

H1,3

NK-I

_

_

HyperforinHypericum

Amentoflavon

HyperforinHypericum

Hypericum

Estrogen

13,118-Biapigenin

Hypericin

Therapy with willow bark extract in comparison with Aspirin

n Indication: osteo-arthritis of the hip and knee,feverish rheumatic disease,low back pain

n Recommended dosage: 1400 mg willow bark extract(ESCOP) @ 240 mg salicylalcohol-derivatives calc. as salicin

Prof. Dr. Dr. h.c.mult. Hildebert Wagner – Center of Pharma Research – Department of Pharmacy – University of Munich Folie 16

derivatives calc. as salicin

n Pharmacokinetic studies: 240 mg salicin ð 10 µMol/l salicylicacid; 1000 mg ASP ð 500 µMol/lsalicylic acid

n Clinical trials: Chrubasik et al. JAMA 109:9-14 (2000)Schmidt et al. Phytother. Res. 15:344-350 (2001) Bieger et al. J. Rheumatol. 31:212-213 (2004)

Synergism between herbal andother drugs I

Synergistic effects of Vitis vinifera seeds (grape seed extract = GSE) with amphotericin B (Amp) against disseminated Candidiasis in mice

Surv

ivors

2

3

4

5

11.4 ± 3.214.4' ± 2.617.6' ± 7.338.4 ± 8.0

MST (Days)

Prof. Dr. Dr. h.c.mult. Hildebert Wagner – Center of Pharma Research – Department of Pharmacy – University of Munich Folie 17

Ref. Yongmoon Han Phytomedicine 2007, in press

n Combination of grape seed extract with Amp. B results in a more than 75% reduction of Amp. B

n The MST value of the mice group which received the combination was greater than MST value from mice group given four times Amp. B dose of 2mg/kg bw.

Results

Days

Surv

ivors

0

1

4010 3020

DPBSAmp aloneGSE aloneAmp plus GSE

healthy tissueimmunostimulants

inhibitors of

Examples of multitarget therapy

Example 1: Cancer therapy

Prof. Dr. Dr. h.c.mult. Hildebert Wagner – Center of Pharma Research – Department of Pharmacy – University of Munich Folie 18

cancer cells

cytostatics

inducers of apoptosis

inhibitors of angiogenesis

stimulants ofoncogen-suppressor

genes

Progress in the field of synergy research and omic technology can

Conclusion

Prof. Dr. Dr. h.c.mult. Hildebert Wagner – Center of Pharma Research – Department of Pharmacy – University of Munich Folie 19

research and omic technology can give phytotherapy a new legitimacy and the possibility to treat diseases which up to now were reserved for

chemotherapy only