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International Workshop on metabolomics in the context of systems
biology: a rational approach to search for lead molecules from nature.
São Paulo, 24-25 February Brazil
Brazilian biodiversity as a source
of new drugs
Emerson F. Queiroz
Department of Research and Development
Aché Laboratórios Farmacêuticos, São Paulo, Brasil
14 %
10 %3%
12 %
5 %
Derivated natural
Products (semisynthesis)
23%
Total synthesis
33%
Total Synthesis
Based in natural product
(Mod. of an existen agent)Vaccine
Total Synthesis
using natural product
pharmacophore
Natural product
Biological
New chemical entities, 1981-2002.
Newman, D. J., Cragg, G. M. and Snader, K. M. (2003). J. Nat. Prod., 66, 1022.
NP are 52%
of the all
chemical
entities !!
Exenatide (INN, marketed as Byetta) is one of a
new class of medications (incretin mimetics)
approved (Apr 2005) for the treatment of
diabetes mellitus type 2)
The Gila Monster and the development of Byetta®
D. Drucker, J. Buse, K. Taylor, D. Kendall, M. Trautmann, D. Zhuang, L. Porter. The Lancet, Volume
372, Pages 1240-1250
Heloderma suspectum (Helodermatidae) 39-amino-acid peptide
The Cone snail and the development of Prialt®
Prommer E (2006). "Ziconotide: a new option for refractory pain". Drugs Today 42 (6): 369–78
Ziconotide
Sirolimus (INN) is a relatively new immunosuppressant drug used to prevent rejection in
organ transplantation, and is especially useful in kidney transplants.
Rapamycin
The development of Rapamune® (sirolimus®)
from Streptomyces hygroscopicus
Rapa Nui Streptomyces hygroscopicus
C. Vézina et al., J. Antibiot. 28, 721 (1975)
S. Sehgal et al., J. Antibiot. 28, 727 (1975)
Tiotropium bromide an anticholinergic drug for the
treatment of chronic obstructive pulmonary disease
Kato M, Komamura K, Kitakaze M (December 2006). "Tiotropium, a novel muscarinic M3 receptor antagonist, improved
symptoms of chronic obstructive pulmonary disease complicated by chronic heart failure". Circ. J. 70 (12): 1658–60.
Atropa belladonna L., Solonaceae
Reconsidering natural products
Pure active constituent(s)Extract(s) Toxicology
Structure
modification
Bio-assay
FractionsMedicinal plants
in vivo assays in vitro assays
HPLC/UV/MS
HPLC/UV/RMN
Chemical screening
O
OH 3 C O
H O
H OO C H 3
H O
O H
O
OO
O
synthesis
structure elucidation
Contemporary methodological approaches in the search for new
lead compounds from higher plants.
Find a Needle in the Haytack
Pedro Álvares Cabral (c.1467-c.1520)
The discovery of Brazil
Brazil was first "discovered" by Europeans
On April 22, 1500.
The Captivity of Hans Staden of Hesse, in A.D. 1547-1555, Among the Wild Tribes of Eastern Brazil in Marburg in 1557.
The early explorers
The Captivity of Hans Staden of Hesse, in A.D. 1547-1555,
Among the Wild Tribes of Eastern Brazil in Marburg in 1557.
The origin of the name “Brazil”
Brazilwood : Caesalpinia echinata Lam. (Leguminosiae)
The source of a perfect red dye
Bueno, E. (1998) Náufragos, Traficantes e Degredados: As Primeiras Expedições ao Brasil, Coleção Terra Brasilis Vol II, Cap. 3, A Terra do Brasil.
Brazilwood : Caesalpinia echinata Lam. (Leguminoseae)
Sir Robert Robinson
The Nobel Prize in Chemistry 1947
Holme, J.; Developments in the Chemistry and Technology of Organic Dyes; J. Griffiths Ed.; Blackwell Scient. Publ.; Oxford, 1984
“the discovery of brazilwood pigment”
Brazil and the Biodiversity
Brazil has 1/5 of the world
biodiversity, with 50 000 species of
plants, 5 000 of vertebrates, 10-15
millions of insects, and millions of
microorganisms.
Brazilian’s Ecosystems
http://www.biodiversityhotspots.org
http://www.biodiversityhotspots.org
Pilocarpine from Pilocarpus ssp. (Rutaceae)
Pilocarpine
Rosin, A. Pilocarpine. A miotic of choice in the treatment of glaucoma
has passed 110 years of use. Oftalmologia. 1991, 35(1):53-5
Cavalheiro E. A. I. J Neurol Sci, 16:33-37, 1995
The Pilocarpine model of epilepsy
Tubocurarin from Amazonia « curare »
Chondrodendron tomentosum
(Menispermaceae)
D-TUBOCURARIN
Curare and its successors.A 50-year's evolution Buzello W, Diefenbach C.Anasthesiol Intensivmed Notfallmed Schmerzther. 1992, 27(5):290
«The introduction of tubocurarine
into anaesthetic practice, by Griffith
and Johnson,in 1942, caused
profound changes in the efficacy
and safety of anaesthesiology »
N
H3CO
CH3
CH3
OHO
N
OCH3
H3C
H3C
O
HO
"the most fruitful basis
for the discovery of a
new drug is to start
with an old drug".
Prof. James W. Black
Nobel Price of medicine in 1988
N
H3CO
CH3
CH3
OHO
N
OCH3
H3C
H3C
O
HO
D-TUBOCURARIN
Atracurium a synthetic derivative based in the structure of Tubocurarin
N
H3COCH3
OCH3
H3CO
H3CO
O O
O O
N
OCH3H3C
OCH3
OCH3
OCH3
ATRACURIUM
(TRACRIUM®)
Conner, C.S. Atracurium and vecuronium: two unique neuromuscular blocking agents.
Drug Intell Clin Pharm. 1984 Sep;18(9):714-6
Bothrops jararaca snake and captropril development
Prof. Sergio Henrique Ferreira
angiotensin-converting enzyme inhibitor
N
HO
OHN
NH
HN
H2N
O
O
NH
O
NH2
O
OHO
N
HO
O
HS
O
N
HO
OHN
O
OO
BPP5
Captopril®
Enalopril®
The stevioside from Stevia rebaudiana Bertoni (Asteraceae)
Crammer, B, and Ikan, R (1986) Sweet glycosides from the stevia plant. Chemistry in Britain 22: 915-916
- 250–300 times sweeter than sucrose
- Heat stable, pH stable, and non-
fermentable.
“Guaraná”
“Açaí”
Paullinia cupana Kunth (Sapindaceae)
Euterpe oleracea Mart (Arecaceae)
Brazilian fruits with high economic value
“Cajá”
Spondias mombin L.(Anarcadeaceae)
Spondias tuberosa Arr. (Anarcadeaceae)
“Umbu”
“Pitanga”
Eugenia uniflora L.(Myrtaceae)
Myrciaria jaboticaba (Vell.) Berg, (Myrtaceae_
“Jaboticaba”
Feijoa sellowiana Berg., Myrtaceae
Review on chemical composition and constituents of some important exotic fruits. Z Lebensm Unters
Forsch. 1981;173(1):47-60
Brazilian Pharmaceutical Industry
Brazilian Pharmaceutical Industry
Brazilian Pharmaceutical Industry
Hydrochoerus hydrochaeris, Capiviidae
The Especifico Pessoa: An Antidote againstBothrops atrox snake venon
Bothops atrox
The Especifico Pessoa: An Antidote againstBothrops atrox snake venon
Prof. Koji Nakanishi
Columbian University
The active compounds isolated fromEspecifico pessoa
Cabenegrin A-I
Cabenegrin A-II
• Injection of 2.5mg/kg venon to 9kg
beagle dog leads to hypothention,
respiratory and cardiac arrest.
•Injection of 1 mg/kg of A-I, 15 min
before or after venon injection
restoures or reverses effect in 90 min.
Nakagawa et al., 1982.Tetrahedron letters, 3855-3858, 1982.
El total de queimadas en la región amazónica
en el periodo de 1999-2005 (km2)
Fuente : Instituto de pesquisa ambiental da Amazônia
The arrival of soy bean in Amazon
The Inovation Law
Ministry of Science and TechnologySecretary of Policy and programs
On research and development
Brasília-2006
The Brazilian Government Initiative
BRAZIL has a Great Potential in Research
Masterdegree and
PhD per year
1987 1988 19890
2.000
4.000
6.000
8.000
10.000
12.000
14.000
16.000
18.000
20.000
1990 1991 1992 1993 1994 1995 1996 1997 1998 1999 2000
Master degree PhD
2001
10 thousand PhD’s annually
Fonte: Capes / MEC
0
2.000
4.000
6.000
8.000
10.000
12.000
81 82 83 84 85 86 87 88 89 90 91 92 93 94 95 96 97 98 99 200020010,00
0,20
0,40
0,60
0,80
1,00
1,20
1,40
1,60
The Brazilian scientific production
represents 1.8% of the word
production
(17º position in 2005)
Fonte: CNPq / MCT
Number of articles
published
in international
journals
Ph.D researchers
University x Company
Aché Laboratories
• The biggest pharmaceutical company in Latin America
• Benefices 2005: U$ 800 millions (Ethics synthetic lines and OTC)
• production : 150 millions units/year
• Number of workers : 5.000
• Sales force : 1,200 representatives visiting 130,000 physicians / month
Modern drug production
Aché’s Laboratory Capacity
Products:
1. Solids ( 72 millions units/year)
2. Liquids (60 millions units/year)
3. Semisolids (12 millions units/year)
Aché’s R&D Pipeline
Projects Indication Preclinical
Clinical Essays
Phase II
Pharmaceutical
form
development
Anvisa
approval MarketingPhase I Phase III
ACH 02 Anti-inflammatory
*ACH 06 Anxiolytic and sedative
ACH 04 Anti-diarrhea
ACH 05 Cognitive disorder
ACH 09 Metabolic syndrome
ACH 22 Hepatic steatosis
ACH 01 Anti-inflammatory
ACH 11 Antifungal
ACH 16 Diabetes Mellitus
ACH 17 Erectile dysfunction
ACH 24 Vitiligo
ACH22 Hepatic esteatosis
Aché partners
UFRGS
UFSC
UERJ
UNB
UFPB
UNICAMP
CPQBA
Nanocore
UNIFESPUNESP
UNICAMP
BIOAGRI
INCOR
Centroflora
UFCE
UFRJ
UFPA
University of Geneva, Switzerland
University of Barcelona, Spain
Acheflan®
The first drug developed in Brazil
Cordia Verbenacea D.C., Boraginaceae
C. Verbenaceae could be found in the
Atlantic rain forest
1 8 9 105 6 72 3 4
11 12 13 14
Zingiberene
-copaene -humulene
trans-cariophylene
-cadinene
bisabolol
-cubebene
-elemene
Fraction 1
(monoterpenes)
GC/MS analysis of C. verbenaceae essential oil
Effect of systemic treatment with α-humulene and trans-caryophyllene
from Cordia verbenacea on bradykinin (BK)-induced paw oedema in miceP
aw
vo
lum
e
(l)
Time after injection (min)
BK (3 nmol/paw)
Humulene (50 mg/kg, p.o., 1 h) BK (3 nmol/paw)
Caryophyllene (50 mg/kg, p.o., 1 h)
Calixto et coll., Journal of ethnopharmacology, 110, 323-333 (2007).
Effect of systemic treatment with α-humulene and trans-caryophyllene from Cordia
verbenacea on platelet-activating factor (PAF)-induced paw oedema in miceP
aw
vo
lum
e (
l)
Time after injection (min)
PAF (3 nmol/paw)
Humulene (50 mg/kg, p.o., 1 h) PAF (3 nmol/paw)
Caryophyllene (50 mg/kg, p.o., 1 h)
Calixto et coll., Journal of ethnopharmacology, 110, 323-333 (2007).
Acheflan’s industrial production
Selection of plants Industrial collection
Extraction of the essential oil
Cultivation
- Formulation
- Quality control, ...
Brazilian Market of Topical Anti-inflammatory Drugs 2003-2007
0,0%
5,0%
10,0%
15,0%
20,0%
25,0%
30,0%
35,0%
40,0%
45,0%
Acheflan® Aché lab. 0,0% 0,0% 7,1% 27,9% 40,7%Cataflam® Novartis 15,7% 14,9% 18,5% 15,7% 12,9%Biofenacaero® Aché lab. 26,4% 27,5% 21,0% 18,6% 14,3%Scaflamgel® Mantecorp 6,3% 6,8% 7,1% 6,6% 6,6%Feldenegel® Pfaizer 7,4% 6,6% 4,5% 4,0% 2,7%Profenidgel® S. Aventis 3,9% 4,0% 4,3% 3,6% 3,7%Generics” 3,1% 4,2% 8,0% 6,1% 5,7%
2003 2004 2005 2006 2007
Med
ical
pre
sc
rip
tio
n
Acheflan ®
Year0,0%
5,0%
10,0%
15,0%
20,0%
25,0%
30,0%
35,0%
40,0%
45,0%
Acheflan® Aché lab. 0,0% 0,0% 7,1% 27,9%Cataflam® Novartis 15,7% 14,9% 18,5% 15,7% 12,9%Biofenacaero® Aché lab. 26,4% 27,5% 21,0% 18,6% 14,3%Scaflamgel® Mantecorp 6,3% 6,8% 7,1% 6,6% 6,6%Feldenegel® Pfaizer 7,4% 6,6% 4,5% 4,0% 2,7%Profenidgel® S. Aventis 3,9% 4,0% 4,3% 3,6% 3,7%Generics” 3,1% 4,2% 8,0% 6,1% 5,7%
2003 2004 2005 2006 2007
Med
ical
pre
sc
rip
tio
n
Acheflan ®
Year
Acheflan® sales in 2009
Acheflan® has 49% of the medical prescriptions
sales 80.000 unit monthly
Cordia verbenacea PROJECT
New indications and formulations:
Oral
Path
Osteoarthritis
Dermatological affection
The "Passion" in "passion flower" does
not refer to sex and love however, but to
the passion of Jesus Christ. In the 15th
and 16th centuries, Spanish Christian
missionaries adopted the unique physical
structures of this plant, particularly the
numbers of its various flower parts, as
symbols of the last days of Jesus Christ
and especially the Crucifixion.
MARZELL, HEINRICH (1927): Deutsches Wörterbuch der Pflanzennamen ["German Plant Name Dictionary"].
Leipzig.
Passiflora incarnata L. (Passifloraceae)
Passiflora incarnata L. (Passifloraceae) and the first world war
During the first wolrld war the preparations with Passiflora was used in
soldiers to treat the shellshock-combat stress reaction.
MARZELL, HEINRICH (1927): Deutsches Wörterbuch der Pflanzennamen ["German Plant Name Dictionary"]. Leipzig.
Passiflora incarnata L.
Strategy for development of a new
Passiflora incarnata L. extract
C-glicosilados flavonoids
Indolic alcaloids
Classical extraction
method
Passiflora incarnata L.
7% of total
flavonoids
Formulation
Extraction
method*
Purification
Method*
*methodology has
been patent by Aché
Strategy for development of a new
Passiflora incarnata L. extract
Phytochemical analysis C-glicosylated flavonoids
254 nm
1 23 6
4
58
7
min0 5 10 15 20 25 30 35
mAU
0
50
100
150
200
Pharmacological assays
21 mg of
flavonoids/caps
Standarization of the ACH6 extract
254 nm
12
36
4
58
7
min0 5 10 15 20 25 30 35
mAU
0
50
100
150
200
nm200 250 300 350 400 450
mAU
0
10
20
30
40
50
60
DAD1, 8.009 (72.4 mAU, - ) of PASSILC1.D
nm200 250 300 350 400 450
mAU
0
10
20
30
40
50
60
70
80
DAD1, 11.156 (87.4 mAU,Apx) of PASSILC1.D
nm200 250 300 350 400 450
mAU
0
20
40
60
80
100
120
140
160
DAD1, 17.010 (165 mAU, - ) of PASSILC1.D
nm200 250 300 350 400 450
mAU
0
20
40
60
80
100
120
140
DAD1, 17.490 (155 mAU, - ) of PASSILC1.D
nm200 250 300 350 400 450
mAU
0
50
100
150
200
250
300
350
DAD1, 20.676 (364 mAU, - ) of PASSILC1.D
nm200 250 300 350 400 450
mAU
0
25
50
75
100
125
150
175
DAD1, 22.649 (193 mAU, - ) of PASSILC1.D
nm200 250 300 350 400 450
mAU
0
20
40
60
80
100
120
140
160
DAD1, 23.996 (168 mAU,Apx) of PASSILC1.D
nm200 250 300 350 400 450
mAU
0
50
100
150
200
250
300
350
400
DAD1, 27.103 (439 mAU, - ) of PASSILC1.D
1 2 3
6 7
4
5 8
ACH06 extract
LC/UV/DAD analysis of the extract of Passiflora incarnata
Uv of flavonoids
Phytochemical study of the ACH06 extract
254 nm
1 23 6
4
5 8
7
min0 5 10 15 20 25 30 35
mAU
0
50
100
150
200
Extrato a 7%
de flav. totais
Semiprep-HPLC
RMN 1H
RMN 13C
Isolamento
RMN 1D e 2D
RMN 500 MHz
Chemical Formula: C21H20O10
Molecular Weight: 432,38
Isovitexin
Compounds identified from the ACH06 extract
254 nm
1 23 64
58
7
min0 5 10 15 20 25 30 35
mAU
0
50
100
150
200
vitexin Isovitexin isoschafitoside schafitoside
isovitexin-2’’-O--
sophoroseisorientin-2’’-O--
sophorose
saponarine vicenina 2’
Thank you for
your attention
Prof. Bejamin Gilbert
Prof. Walter Mors
Prof. Otto Gottilieb
