This document summarizes research on berberine derivatives with antiproliferative activity. Berberine is an alkaloid extracted from plants that has been used in traditional Chinese and Ayurvedic medicine. The researchers synthesized novel berberine derivatives with aromatic groups in the 13 position of the alkaloid skeleton. These derivatives showed improved binding to DNA and greater antiproliferative effects against cancer cell lines compared to berberine. Certain derivatives displayed promising activity against mesothelioma and liver cancer cell lines that warrants further preclinical development studies. The results support berberine's role in inhibiting cancer cell proliferation and its potential as a lead compound for anticancer drug discovery when modified with aromatic groups.

1. Novel 13-(di)arylalkyl berberines with
antiproliferative activity
Gaetano Fiorillo, Franco Buzzetti,
Paolo Lombardi, Tanjia Monir Syeda
via G. Di Vittorio 70, 20026 Novate Milanese, Milano, Italy
Email: staff@naxospharma.eu
Rimini 27-29 October 2015

2. Berberine
Background History
Bitter-tasting isoquinoline quaternary alkaloid extracted
from plants of the genus Berberis, Coptis and others.
In use in the Ayurvedic and Chinese medicines since
hundreds of years.
It shows diverse pharmacological activities:
 Anti-microbial/parasitic,
 Anti-diarrheal, anti-inflammatory,
 Anti-arryhthmic,
 Cholesterol-lowering
 Anticancer

3. The precise molecular basis of its many biological
activities are still debated
Modulation of protein expression by interaction with
nucleic acids is postulated
The interactions between berberine and nucleic acids,
reported since 1962, could lead to its anticancer effect
Mazzini, S. et al, Bioorg Med Chem,
2003, 505–514
(NMR Studies)
Ferraroni, M. et al, Chem. Commun.
2011, 4917-4919
(RX studies)
intercalation
minor groove binding
Berberine
DNA Interaction

4. Berberine represents an interesting and
attractive natural lead compound
Chemical modifications might select more
specific medical indications resulting in
derivatives with better (or different)
biological effects compared to the parent
berberine
Performing rational chemical modifications of
berberine structure led to a new class of
derivatives with antitumour properties
Chemical
Programme
Berberin
e

5. Berberine
Anticancer Properties

6. Since aromatic interactions are ubiquitous in nature, and their geometry is relevant for
the molecular recognition in biological system, that could result in better (or different)
biological effects with respect to the parent Berberine
Chemical
Programme

7. from very low to low yields - better with activated halides or
iodides - berberine back from loss of acetone major by-product
Alkylation of enamine (7,8-dihydroberberine)
Berberine derivatives
Synthetic Methods
from low to moderate yields -
berberine and tetrahydroberberine
from disproportionation of enamine
as major by-products

8. generally from good to very good yields
Uncommon aldehyde-enamine condensation1,2
2 Iwasa, K, et al., Planta Medica, 1997, 1961 Cook, AG, Enamines Synthesis, Structure and reaction, 1988, pag 200-201
Berberine derivatives
Synthetic Methods

9. 1) Commercially available aldehydes
2) Commercially available alcohol followed by oxidation
(PCC or TEMPO)
3) Homologation starting from aldehydes 1) & 2)
Of course, most of starting materials, reagents, solvents,
and disposables are from
Berberine Derivatives
Aldehyde Intermediates

10. Another route was used
Berberine Derivatives
Aldehyde Intermediates

11. Berberine
Derivatives
Binding to DNA
1.77
0.35
2.11
11.01
7.6 7.58
6.8
0
2
4
6
8
10
12
Kix10-5(M-1)
Binding costants of NAXs 1
1.77
0.48 0.51
7.07
10.04
8.90
7.48
0
2
4
6
8
10
12
Kix10-5(M-1)
Binding costants of NAXs2
1D. Bhowmik, M. Hossain, F. Buzzetti, R. D’Auria, P. Lombardi, G.S.Kumar, J. Phys. Chem. B, 2012, 116, 2314−24.
2D. Bhowmik, F. Buzzetti, G. Fiorillo, F. Orzi, T. Syeda Monir, P. Lombardi, G.S. Kumar, Med. Chem. Comm., 2014, 5, 226-31.
n = 3
n = 4

12. Berberine
Derivatives
Binding to DNA
0
2
4
6
8
10
12
Kix10-5(M-1)
Pyridylalkyl derivatives
S. Chatterjee, S. Mallick, F. Buzzetti, G. Fiorillo, T. M. Syeda, Paolo Lombardi, K. Das Saha, G. S. Kumar,
RCS Adv., 2015, 5, 90632
n = 5

13. Berberine Derivatives
Antiproliferative effects
STO, MESOII = peritoneal mesothelima cell lines
MSTO = pleural mesothelioma cell line
0
1
2
3
4
5
6
7
8
9
IC50[mM]
Mesothelioma cell lines
STO
MESOII
MSTO
0
1
2
3
4
5
6
7
8
9
IC50[mM]
Mesothelioma cell lines
STO
MESOII
MSTO

14. Berberine Derivatives
Antiproliferative effects
Human breast adenocarcinoma
cell line
Human liver hepatocellular
carcinoma cell line
S. Chatterjee, S. Mallick, F. Buzzetti, G. Fiorillo, T. M. Syeda, Paolo Lombardi, K. Das Saha, G. S. Kumar,
RCS Adv., 2015, 5, 90632
In the Chinese traditionalmedicine,
Berberine has been used for the
treatment of hepatocellular carcinoma;
Berberine derivatives have an in vitro
better effect.

15. Conclusions
 The promising data obtained on relevant cancer cell lines, shown here and
in other scientific reports, support an active role of BBR in inhibiting cancer cell
proliferation.
 To improve this relevant property, many derivatives (essentially with
(hetero)aromatic groups in the position 13 of the alkaloid skeleton) have been
designed and synthesized.
 In general, derivatives proved to be more efficient than the lead compound,
thus opening new perspectives for drug discovery
 Work performed so far has led to the identification of some candidate
compounds with a profile that would justify the progression to late preclinical
development studies.
Aknowledgements: Financial supports were provided by Ministero dello
Sviluppo Economico (Grant. 01709 ) under the 6th call of the EuroTransBio
initiative, transnational project BERTA (BERberine as antiTumour Agents)