"Advanced Topics in Natural Product "

Massuo J. Kato (IQUSP) (QFL-5972)

Goals

To introduce the fundamentals of natural products chemistry

including biosynthetic origin, evolutionary role,
biological functions and classic and current tools for the
study of natural products in several aspects.
 Discovery of first organic compounds
HO

Atropine
O
(Atropa belladona)
NMe
Mein, 1831.
HO
Quinine
(Cinchona)
Morphine Pelletier & Magendie, 1820
(Papaver somniferum)
Sertürner, 1805
Scopolamine
(Hyoscyamus niger)
Cocaine Landenburg, 1881.
(Erythroxylum coca)
Wöhler, 1859
Papaverine
Merck, 1848.

Tubocurarine
Ephedrine (Ephedra sinica) Insulin (pancreas) (Chondrodendron tomentosum)
Nagai, 1885. Abel, 1929. Boehm , 1895.
 Advanced Topics in Natural Products: main topics

Definitions
and theories on
evolutionary role

Economical
Chemical Natural Products importance
ecology throughout history

Methods for Bioprospecting

isolation and
analysis Biosynthetic
pathways
"Advanced Topics in Natural Products (QFL-5972)"

Atividades

4 h/semana:
2 h de apresentações
Busca na literatura sobre temas em discussão .

Avaliações
1) Discussão de temas

2) Redação de um review

2) Elaborar material educativo (textos, videos etc)

Advanced Topics in Natural Products (QFL-5972 – 2022; Tentative program)

Dates Subject and activities Speaker

(14:00-18:00 h)
12 September Introduction MJK

19 Polyketides MJK

26 Shikimic acid, phenylpropanoids and lignoids MJK

31 Terpenoids MJK
03 October Alkaloids MJK
10 Evaluation
17 Chemical ecology
24 Metabolome tools
31 Tools in bioprospecting
07 November Plant transcriptome to unravel biosynthetic pathways Leandro F. de Oliveira (IB-USP)
14 Synthetic biology Joaquim Vogt Marques
(GinkgoBioworks)
21 Discussion  
28 Evaluation  
 Grupo de pesquisa
Diversidade Molecular de Produtos Naturais
dgp.cnpq.br/

Linhas de pesquisas:
Química de Produtos Naturais.
Isolamento, identificação, bioatividade e variabilidade química de
metabólitos secundários.

Biossíntese de metabólitos secundários

Ecologia química e Interações multitróficas

Filogenia molecular de espécies de Piperales

LABORATÓRIO DE QUÍMICA DE PRODUTOS NATURAIS

INSTITUTO DE QUÍMICA

UNIVERSIDADE DE SÃO PAULO

Fundação 1967 (FAPESP)

Otto R. Gottlieb
Massayoshi Yoshida, Nidia F. Roque,
Zenaide S. Ferreira,
Marden A. Alvarenga, Mario Motidome.

Vicente de P. Emerenciano
Massuo J. Kato
7
 Otto R. Gottlieb – um naturalista

Massuo Jorge Kato - IQUSP

15 de outubro de 2020
BIOTA FAPESP
CENTENÁRIO DE OTTO R. GOTTLIEB

Quimiossistemática e Evolução Micromolecular  Vale do São Francisco, 1958

 Origens e influências

Otto R. Gottlieb (1920-2011)

https://www.youtube.com/watch?v=UyOlO-XHfj0&app=desktop
Early phytochemical studies on Brazilian Lauraceae species

anibine 4-methoxyparacotoin

MORS WB, GOTTLIEB OR, DJERASSI C (1957) The chemistry of rosewood -

isolation and structure of anibine and 4-methoxyparacotoin. JACS 79 , 4507.

GOTTLIEB OR, MORS WB (1958) The chemistry of rosewood. 2. Isolation and

identification of cotoin and pinocembrin. JACS 80, 2263.

GONÇALVES NB, CORREA JC, GOTTLIEB OR (1958) Analeptic action of anibine.

Nature 182, 938.

GOTTLIEB OR, MAGALHAES MT (1958) Isolation of piperonylic

acid from Ocotea pretiosa. Nature 182, 742.
 10 years postponing the “Modern-phytochemistry”
Age in Brazil
Academic career
1945: Otto Gottlieb, major in chemistry at Universidade do Brasil
1955: Otto Gottlieb joined IQA (fellowship from CNPq); early work with Aniba rosiodora

10 years working in his grandfather’s essential

oils company, Ornstein & Co - major business was
the exportation of rosewood oil to Europe.
Otto Richard Gottlieb no IQUSP: 1967-1990
(277 artigos em 23 anos)

1990

Massayoshi Yoshida – fotoquímico e fitoquímico;

especialista em elucidação estrutural de lignoides;
66 artigos com Otto Gottlieb.
 Some early steps in the evolution of
Phytochemistry in Brazil

1955: Walter Mors and Carl Djerassi; Introduction of IR; dawn of modern organic chemistry

1958: IQA, Benjamin Gilbert, Keith Brown Jr, Roderick Barnes

1959-1960: Otto Gottlieb – Weissman Institute; 30 MHz NMR; Introduction of NMR

1962: IQA ended activities (Decreto 1477; october 26th)
 Roberto A. Salmeron
"A Universidade Interrompida:
Brasília 1964-1965",
Ed. UnB, 1999, p. 236-237

1964 – 1965: UnB

Instituto Central de Química:

Profs.: Otto R. Gottlieb (coordenador), Ary Coelho da Silva, Jorge de Oliveira

Meditsch, Mauro Taveira Magalhães
Profs. visitantes:  Luís F. de Carvalho, Jaswant Rai Mahajan e William B. Eyton

Alunos: Afrânio A. Craveiro, Alaíde Braga de Oliveira, Eldenor A. Pimentel,

Geovane Geraldo de Oliveira, Gouvan C. de Magalhães, Hugo Clemente de A.
Pimentel, Jamil Corrêa Mourão, Maria Auxiliadora C. Kaplan, Nídia Cavalcante
Franca, Raimundo Braz Filho, Raimundo G. C. Corrêa e Roberto A. de Lima
 Evolution of phytochemistry in Brazil
U. COSTA RICA UNAL U. PORTO-PORTUGAL
UFAM INPA UFPA UFCE UFMA UFPB
UNIFESP UFSE
UFSP UNESP UFBA
"Química de produtos naturais -
USP-SCar UFRPE 100 anos de Otto Gottlieb"
UFPR UFES 22ª Webinar da Academia
UFSCar UFMG Brasileira de Ciências (ABC)
FCF-RP
"Conhecer para entender: O
IAC IBt FIOCRUZ
mundo a partir do coronavírus".
UFF
USP-LQPN UFRJ-NPPN
(Walter Mors,
Benjamin Gilbert)

Massayoshi Yoshida
Nídia F. Roque
Zenaide S. Ferreira
Mario Motidome
Marden A. Alvarenga
 Schultes and Amazonian Myristicaceae

IDENTIFICATION OF 2 NEW BETA-CARBOLINE ALKALOIDS IN SOUTH AMERICAN HALLUCINOGENIC

PLANTS. Agurell S, Holmsted.B, Lindgren JE, et al. (1968) Biochem. Pharm.,17, 2487.

DE-PLANTIS-TOXICARIIS-E-MUNDO-NOVOTROPICALE-COMMENTATIONES-XIII - FURTHER NOTES ON

VIROLA AS AN ORALLY-ADMINISTERED HALLUCINOGEN.
Schultes RE, Swain T (1976) J. Psychedelic Drugs, 8, 317-324.

15 YEARS OF STUDY OF PSYCHOACTIVE SNUFFS OF SOUTH-AMERICA - 1967-1982 – A REVIEW

SCHULTES RE (1984) J. Ethnopharm., 11, 17-32.
EVOLUTION OF THE IDENTIFICATION OF THE MYRISTICACEOUS
HALLUCINOGENS OF SOUTH-AMERICA

Schultes RE (1979) J. Ethnopharm., 1, 211-239.

Virola surinamensis (Rol). Warb.

O
R
OH
MeO virolina R=H
OMe surinamensina R=OH

OMe

1 1
MeO HO O R R

1 1
R R 1
1 R
R OMe
O OH O HO O
O 1
R
MeO OMe 1
R
OMe OMe 1
R O
OMe

BARATA et al., (1978) Neolignans of Virola surinamensis. Phytochemistry,17, 783.

 Neolignans are
dimers of allyl/propenyl
phenols !!!

Otto Richard Gottlieb

MeO OMe

HO OH
E-isoeugenol eugenol

OMe

O O O
O
burchelin
(Aniba burchelli)
Araújo Lima et al. (1972)
 Química e ecologia química de Myristicaceae
Prof. Dr. Hipólito Ferreira Paulino Filho (IQ-UNESP) (1945-1987)
Ramphastos tucanos Cuvieri

Um tucano ‘disfarçado’
Entre 1979 e 1984, Hipólito
observou na região do rio Guaporé
(Rondônia), um fenômeno raro de
mimetismo entre aves.

Ciência Hoje setembro de 2008

Ramphastos vitellinus Ariel 

Prof. Dr. Hipólito Ferreira

Paulino Neto (IB-UEMG)
Lab Ecologia da Polinização
Isto É, 11/07/1984
https://www.instagram.com/lepec.ecologia/
www.facebook.com/lepececologia
Paulino Filho, H.F. (1985) Ecologia química da família Myristicaceae.
Tese de doutorado (IQUSP) Orientador: Otto R. Gottlieb.
 PROJETO RONDON (Integrar para não entregar)
CNPq (Humaitá, abril-maio de 1980)

Bolsa de desenvolvimento científico e tecnológico regional (DCR)

CNPq (1985-1987)

INPA (INSTITUTO NACIONAL DE PESQUISAS DA AMAZÔNIA) – 1952

Instituição de referência mundial em Biologia Tropical

Biosfera-Atmosfera na Amazônia (LBA),

Programa de Pesquisa em Biodiversidade (PPBio),
Projeto Dinâmica Biológica de Fragmentos Florestais (PDBFF) e
07 Projetos da Rede de Institutos Nacionais de Ciência e Tecnologia – INCT
Fruto maduro de Virola surinamensis
Sementes de Virola (ricos em ácidos miristicos)
 Distribution of secondary compounds
in fruits of Virola elongata

Kato MJ, Yoshida M and Gottlieb OR (1990) Lignoids and arylalkanones from fruits of Virola elongata. Phytochemistry 29:1799-1810.
 Possible compartimentalized biosynthesis of lignans
and neolignans in Virola elongata
Kato MJ, Paulino Fo HF, Yoshida M and Gottlieb OR (1985)
Neolignans from fruits of Virola elongata. Phytochemistry 25:279-280.

O
MeO

MeO
R aryltetralones

O O
O

O
O MeO O R
OH
NH2 O
HO O O Ar
L-phenylalanine E-isoeugenol

Ar
O
MeO
OH furofurans
HO
O
coniferyl alcohol
Ar
O
Kato MJ, Yoshida M and Gottlieb OR (1990) Lignoids and
arylalkanones from fruits of Virola elongata. Ar
Phytochemistry 29:1799-1810. dibenzylbutyrolactones
Seedling chemistry of Virola species
mAbs
HPLC profile in extracts of V. sebifera O O
MeO
I.S.
200 leaves O
OH
MeO
OH
O

1 O 2 O
3 O O
100
1

O
0 OH
O
mAbs
0 10 20
min
3 O
400 O

seeds 2 O

200 I.S. 1 6
O
O

5 O
O
3
0
4 O sesamina

0 10
20
min

Danelutte, A. P., Cavalheiro, A. J., Kato, M. J., 2000. Lignoids in seedlings of Virola sebifera. Phytochemical Analysis 1
Micromolecular Evolution, Systematics and Ecology
- An essay into a Novel Botanical Discipline

Foreword by
Klaus Kubitzki

“It is indeed embarrassing for plant

taxonomists that an outsider has
achieved what systematists
themselves have always claimed as
forming the basis for systematics
and taxonomy:

to bring together in a systematic

fashion the available data from
1982 different fields of study”.
 Otto R. Gottlieb´s scientific contribution
24

1990
541 papers:

1954

2009
221 in Phytochemistry

190

6631 citations
(H= 37)

400 collaborators
 Biblioteca Otto Richard Gottlieb
(~2 mil livros, separatas, documentos, fotos...)
Germoplasm conservation
~100 Piperaceae species (Piper and Peperomia)
few Lauraceae and Myristicaceae species)

31
 Phenylethylamines in cell suspension cultures
of Piper cernuum

O OR
O OH O OMe

OMe
OMe MeO OMe OMe
OH OH
PC-4 R=H
dihydroferulic acid methyl dihydrosinapic acid PC-5 R=Me

OH
O
O
O
cubebin

O
O
Costantin et al. (2000)
 O O- O O- O O- O O- O OMe
PAL/C-4H C3H/OMT F5H/OMT
NH 3+
OMe MeO OMe MeO OMe
OH OH OH OH
L-phenylalanine p-coumarate ferulate
sinapate methyl
dihydrosinapate

-
4CL
O O O OH O OR
NH3+
CCR
OMe OMe
OH NH 2 OH OMe
L-tyrosine CAD dihydroferulic R=H, Me
acid
TYDC OH
OH OH OH
NH 2 dopamine O
O
O
cubebin
? OMe
OH
OH O
tyramine coniferyl alcohol O
CELL CULTURES DIFFERENTIATED PLANT
Biosynthesis of benzofuran neolignans in P. regnellii

OH
HO 1
p-coumaric alcohol
?
HO 2

OH
. OH
.
. O
O O O
4 5 HO
3

O
HO 6
(+)-conocarpan

Benevides et al. (1999); Sartorelli, et al. (2000)

Figure 5: Chiral HPLC separations of (+/-)-conocarpan

Synthetic (+/-)-conocarpan

Natural cis/trans conocarpan

from P. regnellii

Conocarpan enzymatically obtained.

HPLC conditions: Chiracel, OD (250x4.6 mm)

column eluted with hexanes:EtOH (95:5) at flow
rate of 1.0 ml/min and detection at 280 nm.
 Shuttle STS 80
Taxus brevifolia (taxol)
Catharanthus roseus (vincristina e vimblastina

brócolis (estudo da lignificação)

Pinus radiata (estudo da lignificação)
Piper aduncum (dilapiol)
Partition of coniferyl alcohol between O

OH
lignin and dillapiol in Piper aduncum NH2

growing under microgravity L-phenylalanine

OH

HO
coniferyl alcohol
OMe

microgravity

MeO
LIGNIN
HO
eugenol

O
safrol

O OMe

dillapiol OMe
 Linhas de pesquisas no

LQPN

Bioprospeção Biossíntese

Ecologia química
Chemically mediated multi-trophic interaction diversity
across tropical gradients
(FAPESP/NSF, 2014-2021, UNR – USP)

Chemical cues Piperaceae

(South-Central
parasitoids Herbivores Americas) seedlings

Taxonomy/Molecular Phylogeny HPLC-ESI and

1
H NMR data
and analysis
Sequestration/Biotransformations

Specificity of plant-herbivores-parasitoids relationship

40
 Chemically mediated multi-trophic interaction diversity across tropical
gradients
(FAPESP/NSF, 2014-2022, UNR – USP)

IQUSP
Massuo J. Kato Biosciences Institute USP
Lydia F. Yamaguchi Antonio Salatino
Vivian E. Cornelio Maria L. Salatino
Mariana A. Stanton Anary P. Egydio
Dimitre A. Ivanov Eny I.S. Floh, Diego T. Souza,
Andrés Camilo Guerrero Paula Elbl, Sergio A. Vanin
Fabio Miyashiro Diego Demarco
Mariana Ingegneri
Elsie F. Guimarães (JBRJ)

Maysa Furlan (UNESP) and team

USA team
UNICAMP Chris B. Jeffrey, Lee Dyer,
Andre V.L. Freitas (Baku) Angela Smilanich,
Leandro G. Cosmo, Lora Richards,
Patricia A. Machado, Thomas Parchman (UNR)
Andre Rangel Eric Tepe (U. Cincinnatti)
Simeão S. Moraes Tara J. Massad (U Memphis)
Why Piperaceae species?
 Basal Angiosperms

1 1
90
100
75
75-105
4090
2858
2929

~250,000 species
Angiosperm Phylogeny Group

http://www.mobot.org/MOBOT/
43
research/APWeb/
 Amides from Piperaceae species
Piper arboreum (PA) P. corcovadensis (PC) Peperomia oreophila (PO)
P. hispidum (PH) P. tuberculatum (PT)
P. scutifolium (PS)

O O O O
MeO O N O
N N N
H O H H
MeO H O
PT-9 PT-8
R PT-6 (R=H)
OMe PO-2
O PO-2 (R= OMe
O O
O
N
O N N
O H
PS-1 PH-4 H
O
MeO PS-2

O O O
N N O N O N O
O O N
O O O O PAR-1 O PH-3
O OCH3 O PAR-3 PH-2
PH-1
PAR-2

O O O
O O O MeO
MeO
O N O MeO N
N N
MeO O N
O PT-5 MeO
O MeO OMe PT-4
PT-7 O OMe
OMe PT-2
PT-3

Ramos et al., (2019); Cotinguiba et al. (2017); Regasini et al. (2009); Alécio et al., (1998); Navickiene et al., (2000);
Silva et al., (2002); Navickiene et al., (2003); Marques et al., (2009). 44
Eois (Geometridae): 600 estimated species

Piper: ~2000 species

Piper malacophyllum
46
 8/9 Neotropical Eois
clades are specific to
Piper species

Two Eois species are

specific to Peperomia
species

Strutzenberger et al., (2010) Zoologica Scripta 39, 603.

 Relação de especificidade
Piper-Eois-parasitoides

1) Determinar a especificidade das relações entre os parasitoides de Piper-

Eois e a química envolvida;

2) Decifrar as respostas (VOC) emitidas pelas plantas após os danos

causados ​pelos herbívoros e suas implicações para a composição da
comunidade de herbívoros e parasitoides;

3) Investigar processos de biotransformação por herbívoros especialistas

versus generalistas.

4) Caracterizar o padrão de diferenciação química durante a ontogenia e suas

implicações para os herbívoros;

48
 120
556
1064 P. caldense OH
HO
1188 COOH

1201
1485 COOH

1486
14 1489
1525 P. carniconnectivum
12 alata
arifolia 6' O
10 blanda HO 4'

galioides H3CO O 1'

OH 7 2 2'
8 glabella
MeO O
hispidula H3CO 5 4
incana
6 OH O
nitida

i
OH O
obtusifolia
4 OMe
oreophilla HO O
pellucida
2
PGN

0
rhombea P. bowiei OH O
sandersii
sincorana P. permucronatum
-2 tetraphylla
20 18 16 14 12 10 8 6 4 2 0 trineura
ppm urocarpa

Inter- and intraspecific variability

?
Naupactus bipes
Eois spp

Heraclydes brasiliensis

Plant insect
Ontogeny
relationships
 The secondary metabolism for amides-producing
Piper species during ontogey
P. amalago
P. tuberculatum
O O O
MeO O
N N

MeO O
OMe 7
5

λ = 250 Adult λ = 280 Adult

0 100200300400
nm leaves nm leaves
200 400

mAU
mAU
0

Seedling Seedling
100

leaves 400 leaves

mAU
mAU
50

200
0
0

0 5 10 15 20 25 30 35 40 45 0 5 10 15 20 25 30 35 40
Retention time (min) Retention time (min)
 Metabolomic analysis during ontogeny of Piper gaudichaudianum
HO O HO O
HO O

NH2 OMe
MeO

OMe OMe
O
OMe 1a O
OMe
OH O
OH OH O apiole dillapiole
L-phe eugenol
p-coumaric ferulic
acid O acid P. regnellii P. solmsianum P. gaudichaudianum
HO2C HO2C

OH O
O
MeO 2C

O
P. gaudichaudianum
O
conocarpan
HO P. solmsianum

MeO OMe
O
MeO OMe MeO OMe OMe
MeO
OH OMe OMe OMe
P. regnellii grandisin
OH E-isoelemicin

PCA-NMR of
P. gaudichaudianum

O
O O O O
HO2C HO2C
HO HO

OH O [OH
H] O
O
51
2 3a MeO 2C

O
Gaia, A.M., Yamaguchi, L.F., Jeffrey, C.S., and Kato,
O
O M.J. (2014). Age-dependent changes
OH from allylphenol to
P. gaudichaudianum O

prenylated benzoic acid production in Piper gaudichaudianum

HO conocarpan Kunth. Phytochemistry 106, 86-93.
- H 2O HO
 Major ontogenetic changes in secondary compounds
of Piper species
R3 R4
R1O
HO2C O
O R2O
O
HO 15a R=H
15b R=OMe O
6 R
N O
P. gaudichaudianum 7 13a-13d O
22
CO2H O P. richardiaefolium
HO
14 P. regnellii O lignoids
OH O N
9

P. hemmendorffii O OMe
OH
20
CO2H MeO OMe MeO O
O

MeO OMe
P. reticulatum
CO2H 16
HO OMe 12 OMe OH O
OH
P. solmsianum P. permucronatum
11
P. caldense

prenylated benzoic acids lignoids Group II

Adult flavonoids Adult

Group I O Group III

O N
O
21
O
O
R2 O 5 P. amalago O
N
O OMe H N
18
O
P. solmsianum H
R1 O O
OMe O O 18
MeO MeO
MeO N
1 (R1=H; R2= OMe) N
2 (R1=OMe, R2=H) MeO 3 MeO
MeO 17
OMe OMe CO2Me
OMe 17 O
P. gandichaudianum P. regnellii
P. hemmendorffii O O
MeO P. tuberculatum O
23
P. solmsianum N
P. regnellii MeO OMe 4 O 19

P. caldense Seedling P. richardiaefolium P. permucronatum

phenylpropanoids amides amides amides-ester

Seedling Adult Seedling

Gaia, AM, Yamaguchi, LF, Guerrero-Perilla C, Kato MJ (2021) Plants 10(6), 1085 (doi: 10.3390/plants10061085).
52
Secondary compounds during ontogeny of
Peperomia pellucida
MeO O

MeO
OMe O OMe OMe
7

MeO OMe
OMe OMe
5

O OMe
OMe
4

53
 Geometridae

P. pellucida leaves
UV
under different
treatments
jasmonic acid

Monoplatus
54

control and
insects
MeO O

MeO
OMe O
7

OMe OMe

Group I Group II
ØControl MeO OMe
ØUV light
jasmonic acid
Ø Monoplatus
O OMe OMe
Ø Geometridae
ØDrought
5
O OMe ØTemperature
Group III
OMe
4
ØEdessa meditabunda
ØDarnkess UV
ØJasmonic
. acid ØMechanical injury

dillapiol

Moraes MM, Kato MJ (2021) Biosynthesis of Pellucidin A in Peperomia pellucida (L.) HBK.
Frontiers in Plant Science 12 (413). doi:10.3389/fpls.2021.641717
 Biosynthesis of Pellucidin A in Peperomia pellucida

O HO2C CO2H
O
MeO MeO OMe
MeO
OH
MeO OMe
OH
x
HO UV365
ferulic acid MeO OMe
2,4,5-trimethoxycinnamic acid OMe MeO

I -CO2

-CO2

MeO OMe
O MeO

OH
L-Phe MeO OMe
MeO OMe
2,4,5-trimethoxystyrene UV365 OMe MeO
cinnamic acid
pellucidin A

HO OH
O
-CO2 HO
HO
OH
HO OH
x
HO UV365 HO OH
OH HO
caffeic acid 2,4,5-trihydroxystyrene
II

Moraes MM, Kato MJ (2021) Biosynthesis of Pellucidin A in Peperomia pellucida (L.) HBK.
Frontiers in Plant Science 12 (413). doi:10.3389/fpls.2021.641717 55
 Biosynthesis of pellucidin A in P. pellucida

MM 238 MM 194 * MM 388

MeO OMe

O
O * *
MeO OMe
* OH MeO
* MeO
OH * MeO OMe
NH2 OMe MeO
MeO OMe MeO OMe
389 [M+1] +. MeO OMe
L-phenylalanine OMe MeO
2,4,5-trimethoxycinnamic acid 2,4,5-trimethoxystyrene
pellucidin A
390 [M+2] +.

% %
A 100.0 100.0
194 [M]+.
% natural
abundance
238 [M]+.
0.40
75.0 75.0 388 [M]+.
0.30
50.0 50.0
0.20
239 [M+1]+. 389 [M+1]+.
25.0 25.0 195 [M+1] +. 0.10 390 [M+2]+.
0.0 A1 0.0 A2 0.00 A3
m/z m/z m/z

B
% % % 13C-enriched
100.0 100.0
194 [M]+. 0.40
238 [M]+.
75.0 75.0
O 0.30 388 [M]+.
50.0
195 [M+1]+.
50.0 +
* OH 0.20 389 [M+1] .
NH2 239 [M+1] +.
25.0 25.0 0.10 390 [M+2]+.
L-phenylalanine 0.0 B1 0.0 B2 0.00 B3
m/z m/z m/z

%
C 100.0 % 13C-enriched
194 [M]+. 0.40
75.0
195 [M+1]+. 0.30 389 [M+1]+.
50.0
0.20 [M] +. 56
25.0
388
0.10 390 [M+2] +.
 MM 238 MM 194 * MM 388
MeO OMe

O
O * *
MeO OMe
* OH MeO
* MeO
OH * MeO OMe
NH2 OMe MeO
MeO OMe MeO OMe
389 [M+1] +. MeO OMe
L-phenylalanine OMe MeO
2,4,5-trimethoxycinnamic acid 2,4,5-trimethoxystyrene
pellucidin A
390 [M+2] +.

% %
A 100.0 100.0
194 [M]+.
% natural
abundance
238 [M] +.
0.40
75.0 75.0 388 [M]+.
0.30
50.0 50.0
0.20
239 [M+1]+. 389 [M+1]+.
25.0 25.0 195 [M+1] +. 0.10 390 [M+2]+.
0.0 A1 0.0 A2 0.00 A3
m/z m/z m/z

B
% % % 13C-enriched
100.0 100.0
194 [M]+. 0.40
238 [M]+.
75.0 75.0
O 0.30 388 [M]+.
50.0
195 [M+1]+.
50.0 +
* OH 0.20 389 [M+1] .
NH2 239 [M+1] +.
25.0 25.0 0.10 390 [M+2]+.
L-phenylalanine 0.0 B1 0.0 B2 0.00 B3
m/z m/z m/z

%
C 100.0 % 13C-enriched
194 [M]+. 0.40
75.0
O
195 [M+1]+. 0.30 389 [M+1]+.

Incorporation
MeO
* OH 50.0
0.20 [M] +.
MeO OMe
25.0 388
0.10 390 [M+2] +.

of 13C-
2,4,5-trimethoxycinnamic acid C3
0.0 C2 0.00
m/z m/z

precursors in D 0.20
%

[M+1]+.
13C-enriched

P. pellucida
0.15
MeO 389 390 [M+2]+.
* 0.10 [M] +.
MeO OMe 388
0.05
2,4,5-trimethoxystyrene
0.00 D3
m/z

Moraes MM, Kato MJ (2021) Biosynthesis of Pellucidin A in Peperomia pellucida (L.) HBK.
57
Frontiers in Plant Science 12 (413). doi:10.3389/fpls.2021.641717
Biosynthetic pathways involved in the formation of major secondary compounds in
Piper species with transcriptome analysis

O
(S)-canadine
PKS; acyl-CoA ligase; acyl-CoA transferase + lysine O Potentially operating
N
O
OMe
Ar
m n N
H in Piper species
R
SCoA
OMe O O O Ar RO O O
BBE R O Ar OH Ar OH Ar
Ar OH CHI PLR Ar
CHS O
O
O O OH O
Ar O Ar O Ar HO Ar
N R
N pinoresinol lariciresinol secoisolariciresinol matairesinol

Ar

H PKS III
Mannich Ar OPP
CO2H
O O P. gaudichaudianum
aristolactam
(via aristolochic acid) geranyl-PP
Ar HO
NH2
OH
O adults
p-hydroxybenzoic acid IGS
via L-tyrosine (?) gaudichaudianic acid
(TAL)
O OMe
OH isoeugenol
O OH O CoA O H OCR OH
O OH O OH

C4H O
PAL C4H CAD CAD
NH2
O OMe
C4L OMT-SAM EGS1
OMe seedlings
OH OH OMe
OMe OMe dillapiol
OH OH OH
OH OH OH
cinnamic coniferyl alcohol eugenol
L-phenylalanine acid p-coumaric acid caffeic acid-CoA caffealdehyde

MeO
MeO MeO P. malacophyllum
Acyl-CoA transferase
(seedling and adult) BBE: berberine bridge enzyme
PKS III O O HO
O alkenylphenols CAD: coniferyl alcohol
OMe piperolides O dehydrogenase
O CES: chavicol/eugenol synthase
CHI: chalcone isomerase
NH2
HO CHS: chalcone sintase
L-lysine O C4H: cinnamoyl-4-hydroxylase
H2N L-lysine O O O
decarboxylase EGS: eugenol synthase
O N O N N N IGS, isoeugenol synthase
O OH H H OH OMT: O-methyltransferase
PKS III O O PAL: phenylalanine ammonia lyase
pirrolidil amides P. arboreum (adult) PLR: pinoresinol–lariciresinol
O reductase
PKS: Polyketide synthase III
NH2 L-ornitine
HO SAM: S-adenosylmethionine
decarboxylase TAL: tyrosine ammonia lyase
O O O O
H2N
MeO O N
L-ornitine N N
O H
MeO
OMe piplartine piperidyl amides P. tuberculatum (adult)
Selection and validation of reference genes for measuring gene expression in
Piper species at different life stages using RT-qPCR analysis

Study of Piper species in two developmental stages

(seedlings and adults) by RNASeq

Oliveira et al., (2021). Plant Physiology and Biochemistry 171, 201-212.

59
 Perspectivas
1) Especificidade: Estudos de eletroantenografia Eois x VOCs de Piper

2) Linguagem quimica (VOCs): Estudos de comportamento e

transcriptoma de antenas dos insetos

3) Ontogenia: Transcriptoma comparada de plântulas x plantas

adultas e ensaios biologicos.

4) Biotransformação e especificidade: requer estudos comparados

entre insetos modelos e os herbivoros especialistas de Piper.

Ecología química como linha de pesquisas:

como funciona a natureza???
60
Por que estudar Produtos Naturais?

Qual a composição química?

Qual a atividade biológica e

relação estrutura-atividade?

Há potencial econômico ?

61
Como e por que buscar novas moléculas
na natureza?

Congresso IQ 2021

Agenda 2030 e o Desenvolvimento Sustentável

Massuo Jorge Kato 62

Objetivos de desenvolvimento sustentável (ODS)
1. Erradicação da pobreza
2. Fome zero e agricultura sustentável
3. Saúde e bem estar
4. Educação de qualidade
5. Igualdade de gênero
6. Água potável e saneamento
7. Energia limpa e acessível
8. Trabalho decente e crescimento econômico
9. industria, inovação e infraestrutura
10. Redução de desigualdades
11. Cidades e comunidades sustentáveis
12. Consumo e produção responsáveis
13. Ação contra a mudança global do clima
14. Vida na água
15. Vida Terrestre
16. Paz, justiça e instituições eficazes
17. Parcerias e meios de implementação 63
 Cenário mundial durante a COP26
• Explosão populacional (7.9 bi em 2021-> 8,9 bi em 2035)

https://www.worldometers.info/world-population/world-population-projections/ 64
 Cenário mundial durante a COP26

• Explosão populacional (7.9 bi em 2021-> 8,9 bi em 2035)

• Devastação florestal e perda da biodiversidade;
• Aumento do consumo de combustíveis fósseis e
aquecimento global;
• Novas moléstias ainda sem curas (SARS, gripe aviária,
corona virus, cancer, etc);
• Aumento da resistencia de microorganismos (MRSA,
VRSA);

[entre os 1556 novos medicamentos aprovados nos últimos 30 anos

pelo FDA, somente 1,3% (20) foram para doenças do terceiro mundo]

65
 Detecção de superbactérias
resistentes a antibióticos triplica no
Brasil durante a pandemia
Em 2019 Fiocruz recebeu mil amostras; em 2021, até outubro, já são mais de
3,7 mil. O processo tem sido acelerado pelo consumo excessivo – e muitas
vezes incorreto – de remédios na pandemia

Roberta Jansen, O Estado de S. Paulo

06 de dezembro de 2021.

66
 Principais causas de mortes no mundo (2016)
56,9 milhões em 2016; 54% resultam dessas 10 causas.
milhões

1. Cardiopatias isquêmicas (estilo de vida) 15,2

2. AVC (acidentes vasculares cerebrais) Covid-19
6,2 6,4 mi
3. Doenças pulmorar obstrutiva crônica (2022)
3,2
4. Infecções das vias respiratórias inferiors 3,0
5. Alzheimer e outras demências 2,0
6. Cancer de pulmão, traqueia e brônquios (85% fumantes) 1,7
7. Diabetes mellitus 1,6
8. Doenças diarreicas 1,5
9. Tuberculose (9 milhões de casos/ano) 1,4
10. Acidentes de trânsito 1,3

Baseado em dados da World Health Organisation (WHO) (2018)

https://www.proclinical.com/blogs/2017-5/the-leading-causes-of-death-in-the-world-can-they-be-cured
 Desenvolvimento de resistência

Antibiótico Ano de Ano em que

introdução se detectou
resistência
Sulfonamidas 1930 1940
Penicilina 1943 1946
Estreptomicina 1943 1959
Cloranfenicol 1947 1959
Tetraciclinas 1948 1953
Eritromicina 1952 1988
Vancomicina 1956 1988
Meticilina 1960 1961
Ampicilina 1961 1973
Cefalosporina anos 60 final de 60
 The Drug Discovery Process

100,000
Samples Extracts

10 Bioassays
Years
(High throughoutput screening)

1
Product
“Bioactive product”
All new approved drugs 1981-2014, by source (N = 1562)

Vaccine large (> 45 residues) peptide or protein

Derived from
natural
Totally products
synthetic
drug

"B": Biological; usually a large (> 45 residues) peptide or protein either isolated from an organism/cell line or
produced by biotechnological means in a surrogate host.
"N": Natural product.
"ND": Derived from a natural product and is usually a semisynthetic modification.
"S": Totally synthetic drug, often found by random screening/modification of an existing agent.
"S*": Made by total synthesis, but the pharmacophore is/was from a natural product.
"V": Vaccine.
"NM": Natural Product Mimic.

Newman and Cragg (2016) Natural Products as Sources of New Drugs

from 1981 to 2014. J. Nat. Prod., 79 (3), pp 629–661.
Medicamentos (40% a base de produtos naturais)

Hipocondria
ou
necessidade
dos tempos
modernos?
 MeO HO AcO
MeO

O O O
O
NMe NMe NMe
NMe
OH
HO HO AcO
O heroine
oxycodone codeine morphine
semi-sintético semi-sintético
(obtido a partir da
tebaina)
natural
z z

Mais de 15 milhões de
pessoas consomem opiáceos
de forma ílicita (opio, morfina
ou heroina).
Mercado mundial de morfina:
9.6 Bilhões de dólares em 2020.

ópio (papoula - Papaver somniferum) Art: André Kato

 Tabaco
Primeira exploração commercial de um produto natural maléfico à saúde humana .

1559 – Embaixador francês em Portugal, Jean Nicot

enviou sementes de tabaco dos Andes.
Sec 17, relatos de usos por ameríndios na Virgínia.
Sec 18, Aborígenes da Austrália usando N. bethamania.
Sec 19, relatos de usos de cigarros de forma social na África
Nicotiana tabacum
Baixo potencial econômico como cigarro:
Planta de fácil adaptação, fácil processamento e armazenamento.
Formas de consumo: cachimbos, mastigação, como rapé.

1660 – 25% dos impostos do Reino Unido oriundos do tabaco (~ 25 % dos homens
fumavam mais de 3 x semana).

1856: Philip Morris

1860 – 1960: Uso de papel para a confeção de cigarettes;

1875 -1880: consumo annual nos EUA, 42 - 500 milhões de cigarros; marketing.
 Mortes por overdose: mais de 750 mil pessoas
morreram por overdose de drogas nos EUA desde
1999.

Aumento de mortes por opioides sintéticos (como o fentanil ilegal),

com alta de 38,4% sobre os 12 meses anteriores.
"O aumento nas mortes por overdose de drogas foi acelerado
pela pandemia de covid-19“.

fentanil carfentanil

100 vezes mais 100 vezes mais

morfina potente que a morfina. potente que o fentanil.

•Alessandra Corrêa
•De Washington (EUA) para a BBC News Brasil, 31 dezembro 2020
https://www.drugabuse.gov/drug-topics/trends-statistics/overdose-death-rates
 The tools:
Spectroscopy
and spectrometric
methods

Metabolomic
profiling

Metabolomics for Functional Genomics,

Systems Biology, and Biotechnology
Saito & Matsuda (2010)
Annu. Rev. Plant Biol. 61:463–89
Additional tools: METABOLOMICS DATA SETS

METLIN
Metabolite & 
MS/MS Database
 General scheme for isolation and
structural elucidation of natural compounds
Fractionation and
Plant tissue, Extraction by Chromatographic
purification
microorganism stem distillation, Pure profile, melting point,
(solvent extractions,
and etc organic solvent compound boiling point,
column chromatography,
or CO2 refraction index, etc)
HSCC, etc)

Determination of
molecular formula
(elemental analysis
or HRMS)

Determination of Determination of
absolute configuration functional groups
(optical rotation, (IR, UV,
circular dichroisms and R-X) 1
H and 13C NMR)

Determination of Determination of types of carbons

tridimensional structures
or spatial relationships
(J3 1H-1H;
13
C NMR: NOESY, TOCSY)
Determination of
(CH3, CH2, CH, C)
connectivities and planar
structures (J3 1H-1H; and sub-structures
13
1
( H NMR: multiplicities and integration;
C NMR: HSQC; HMBC)
13
C NMR: chemical shifts and DEPT 135)
Por que estudar Produtos Naturais?

Qual a composição química?

Qual a atividade biológica e

relação estrutura-atividade?

Há potencial econômico ?

Estudo da
biossíntese
79
 Natural products
(Secondary metabolites)
"Whereas primary metabolites are present in every plant cell
that is capable of division, secondary metabolites are present only
accidentally and are not essential for plant life"
Albrech Kossel (1891)

Some characteristics
Restricted occurrence,
Low molecular mass,
Structurally diverse,
Component of a complex metabolic sequence.
 CO2
+
hv CARBOHYDRATES
H 2O
glycolysis COOH

PHENYLPROPANOIDS
HO OH
piruvic HO O OH
acid shikimic acid
OH

acetate/
malonate
pathway O
FATTY CH3CSCoA
TCA
AMINOACIDS ALKALOIDS
ACIDS acetil-CoA
acetate/
mevalonate
POLYKETIDES pathway PROTEINS
(aliphatic and aromatic) NUCLEIC
ACIDS
OPP
OPP IPP
DMAPP

TERPENOIDS STEROIDS
 Vias biossintéticas

Enzimas
específicas G I
A B C E

Enzimas
inespecíficas
H K

D F
 Variabilidade do metabolismo secundário
Fase de desenvolvimento,
condições edáficas, sazonalidade, etc

100

plântulas % frutos

0 5.00 10.00 15.00 20.00 25.00 30.00 35.00 Time

40.00

Culturas de células
 Variação circadiana da expressão
fenilpropanoídica
em Arabidopsis thaliana

Harmer et al. (2000) Science, 290, 2110.

 Prenylated benzoic acids and chromenes from
Piper aduncum

O O

MeO RO
O O O
OH O
RO
3 MeO
O 5 (R=H)
6 (R=Me)
O 1 (R=H); 2 (R=Me) 4 OH

HO O
O
R1 O
OH RO
OR2 O O
OMe
7 (R1=R2=H )
HO
8 (R1=H; R2=Me) 12 (R=H)
9 (R1=Me, R2=H) 13 (R=Me)
OH
10 (R1=R2=Me)

11

Lago et al., (2004) J. Nat. Prod. 67, 1783.

Baldoqui et al (1999) Phytochemistry, 51, 899.
Parmar et al (1997) Phytochemistry, 46, 597.
 Prenylation of chromenes in Piper aduncum
has circadian rhythm
Content of chromenes

HPLC chromatogram obtained from the assay of

prenyltransferase activity using a protein extract
from leaves of P. aduncum
Prenyltransferase activity
O
O
HO
HO
O
O

1 2

Morandim et al., (2005) Phytochem. Anal., 16, 282.

Classification based on chemical structure

OH OH

thymol geraniol

Thymus vulgaris Primary component of rose

(common thyme) oil, palmarosa oil, and citronella oil.

aromatic compound aliphatic compounds

cyclic compound acyclic compound

 Classification based on
the occurrence (taxonomy)

Ergot alkaloids, and etc

Classification based on the bioactivity

Vitamins Antibiotics

Toxins Allelopathic
Classification based on the biosynthesis

acetate-malonate aliphatic and aromatic

polyketides

isoprenoid C10, C15, C20,

(mevalonate – MEP) C25, 30, 27, C40, few
aromatic, polymers

Shikimate C6-C3, lignans, lignins

Alkaloid-derived phe, tyr, lys, orn-

compounds derived alkaloids

Mixed-origin Flavonoids, alkaloids

 Acetate-malonate pathway
Understanding the biosynthesis (biogenesis)
O
O O O O O O O O
CH3 C SR
-O C CH C SR Claisen
2 OH
OH
acetil-SR (1) malonil-SR (3) O O

O O
O
SCoA aldol
OH
O O HO OH
ácido orselínico

O O OH O
CoAS
Claisen

O O HO OH
floroacetofenona

92
Understanding the biosynthesis (biogenesis)

O
O O O O O O O O
CH3 C SR
-O C CH C SR Claisen
2 OH
OH
acetil-SR (1) malonil-SR (3) O O

O O
O
SCoA aldol
OH
O O HO OH
ácido orselínico

O O OH O
CoAS
Claisen

O O HO OH
floroacetofenona

93
Alguns monoterpenos constituintes
de óleos essenciais

O
O
CHO

cânfora
carvona citronelal

OH

linalol eucaliptol
(óleo de pau-rosa) (1,8-cineol)
 Terpenos:
Sub-classe No de átomos No de Exemplos
de carbono isoprenos  

Constituintes de óleos essenciais como

Monoterpenos 10 2
geraniol, linalol, mentol, etc.

Constituintes de óleo essenciais como farnesol;

Sesquiterpenos 15 3 ácido absisico, lactonas sesquiterpênicas,
hormônios juvenilizantes, piretrinas,
artemisina.
Constituintes de resinas de árvores (ácido
Diterpenos 20 4
abiético); fitol, vit. A, taxol.

Triterpenos 30 6 Esqualeno, trit. pentacíclicos

Esteróides 28 6 Colesterol, testosterona, androsterona,

progesterona, ácido cólico.
Tetraterpenos 40 8 -caroteno (cenoura), licopeno (tomate)

Politerpenos n>40 n>8 Borracha natural, dolicóides

 Encadeamento cabeça-cauda através de reações de SN1
preniltransferases
prenyltransferases are
pirofosfato
all homodimers of
H de isopentenila
70 to 80 kD,
OPP
OPP
require either Mg2+
isomerase
MONOTERPENOS or Mn2+ with a
OPP
pirofosfato
pirofosfato
de geranila
stoichiometry
de 3,3-dimetilalila of two metal ions per
FPP sintase catalytic site.
GGPP
sintase OPP
OPP pirofosfato SESQUITERPENOS
de farnesila
pirofosfato
de geranilgeranila 2x

2x

DITERPENOS TETRATERPENOS

esqualeno

ciclase

HO ESTERÓIDES HO
TRITERPENOS
Envolvimento de sintases nas etapas-chave de
formação de terpenos
 Major biosynthetic steps required for the formation of membrane constituens
(fatty acids and fatty acids and terpenoids by MVA and MEP pathways

MEMBRANE CONSTITUENTS TERPENES

OH OH

OH OH OH STEROIDS
HO
diplopterol tetrahydroxybacteriohopane HO HO TRITERPENES
cholesterol (animals) -sitosterol (plants)
(prokaryotes) (prokaryotes) (C30)
RO

O O GlO

O O
corynexanthin (Gl: glucose) TETRATERPENES squalene
OR glyceroyltetraterpene eter (Archaea)
(prokaryotes) OH 2x (C40)
O 2x
O
O NH4+ O
O P O OPP PT OPP
O O -
phosphatidylethanolamine DITERPENES
O (C20) IPP SESQUITERPENES
(cephalin - major phospholipid in bacteria) farnesyl
O geranylgeranyl (C15)
OR triglycerides (esters, common in animals and plants) diphosphate
OR diphosphate
NH3+ (FPP)
O O + PT (GGPP)
O N(CH3)4 glyceroyl diterpene eters (Archaea) n IPP
O P OH O P O CO2- O P O
R= PT IPP
O - O- O- POLYISOPRENES
phosphatidylcholine
phosphatidic acid phosphatidylserine (lecithin - major phospholipid in animals) OH
(plant and animals)
n m
dolichols (n=2, m=10-16) OPP
geranyl
diphosphate MONOTERPENES
(GPP) (C10)
O FATTY ACIDS
PROSTAGLANDINS 5 8 11 14 n n
LEUCOTRIENES 20 gutta-percha (n= ~1500) rubber (n=10.000-20.000)
HO 1

THROMBOXANES arachidonic acid (C20:4; cis,cis,cis,cis-5,8,11,14)

PROSTACYCLINS O O
prenyltransferases (PT)
Mevalonic acid (MVA) Pathway
HO HO
-linolenic acid (C18:3; cis, cis, cis-9,12,15) palmitoleic acid (C16, cis-9) malonyl CoA
O O O- O O-
O
OH 3,3-dimethylallyl
POLYACETYLENES HO SCoA SCoA diphosphate
HO OPP
linoleic acid (C18:2; cis,cis-9,12) palmitic acid (C16) Aldol (DMAPP)
O O O Claisen
O
O
FAS (fatty acid synthases) HO O
O O OH OH
HO 3-(R)- H isopentenyl
CoA: coenzyme A oleic acid (C18:1 cis-9) myristoleic acid (C14:1; cis-9) SCoA pyrophosphate (IPP)
O SCoA mevalonic acid
ACP: acyl carrier protein O acetoacetyl-SCoA (MVA)
acetyl SCoA OPP
MAT: malonyl/acetyl transferase HO HO
KS: ketosynthase stearic acid (C18) myristic acid (C14)
DH: dehydratase 2-methyl-erythritol-4-phosphate
ER: enol reductase MEP (or DOXP) Pathway
Isoprene pathways
- n x [KS, KR, DH, ER] CO2- H
O O
malonyl-SACP Bacteria MVA or MEP O
O pyruvate
SACP NADPH Archaea MVA HO
NADPH O + OPP
O O O O + H+ NADP+
O OH O + H+
NADP+ O Fungi MVA HO
OPP
MAT O O OH
DH Green algae MEP
CoAS H H+ OH
ACPS KS ACPS ACPS ACPS
KR ACPS
ER Plants MEP and MVA OH 1-deoxyxylulose-5-P 2-methyl-erythritol-4-P (MEP)
acetyl-SCoA acetyl-SACP 3-ketobutyryl-SACP 3-hydroxy-butyryl-SACP butyryl-SACP
butenoyl-SACP Animals MVA glyceraldehyde-3-P (DOXP)
 O
OH
NH2
SHIKIMIC R
R=H phe
ALKALOIDS
R=OH tyr
ACID O
OH
NH2
N
H
tryptophan

O
OH
PHENYLPROPANOID

O
FLAVONOID
O 99
 O O- O O- O O- O O- O O-
O O-
O O-
PAL
NH3+ C4H C3H COMT F5H COMT
? ? ?
OH OMe HO OMe MeO OMe
cinamato OH OH OH OH OH
L-fenilalanina 5-hidroxiferulato
p-cumarato cafeato ferulato sinapato

4CL 4CL
4CL 4CL 4CL

O SCoA O SCoA O SCoA O SCoA O SCoA

M G FLAVONÓIDES,
CCoA3H COMT COMT CUMARINAS,
SUBERINAS,
? AMIDAS
OH OMe HO OMe MeO OMe
OH OH cafeoil-CoA OH OH OH E OUTROS

Phenylpropanoid
4-cumaroil-SCoA feruloil-SCoA 5-hidroxiconiferuloil-SCoA FENILPROPANÓIDES
sinapoil-SCoA
CCR CCR CCR CCR
CCR

pathway O H O H O H O H
A
O H

LIGNINAS
COMT F5H COMT

OH OMe HO OMe MeO OMe

OH OH aldeído OH OH 5-hidroxi- OH
p-cumaraldeído cafeico coniferaldeído coniferaldeído sinapaldeído

CAD CAD ?
CAD ? CAD ? CAD
OH OH OH
OH OH

? COMT F5H COMT

LIGNANAS
HO OMe MeO OMe
OH OMe
OH OH OH OH
OH
álcool p-cumárico álcool
álcool cafeoílico álcool coniferílico álcool sinápico
5-hidroxiconiferílico
? ? ? ?

? ?
NEOLIGNANAS
OMe OMe MeO OMe MeO OMe
OH OH OH OH OH OH
E-isoanol p-hidroxi- E-isoeugenol eugenol E-5-metoxi- 5-metoxi-eugenol
alilbenzeno isoeugenol
 Involvement of p-coumaroyl-CoA in the biosynthesis cinnamoyl
derivatives, following chain extension by PKS enzymes.
O O O OH
malonyl-CoA
SCoA SCoA
PKS
HO HO O HO O OH
p-hydroxybenzoic xanthone (Clusiaceae)
acid

O O O O HO CO2H
HO R
OH SCoA OH OR1 O
HO HO HO caffeic acid HO
p-coumaric acid HO O OH
p-coumaroyl-CoA O
R cinnamoyl esters HO OH
N (R1= aliphatic chain,
-
O O malonyl-CoA H chlorogenic acid (Svetol)
HO constituents of suberins)
(Coffea robusta, Phyllostachys edulis)
PKS O cinnamamides
2x
SCoA

O
OH O O O
SCoA Claisen
HO OH O SCoA OO
O O
styrylpyrones
HO HO HO
(Goniothalamus, Cryptocarya)
PKS malonyl-CoA
O OH OH

O O O O O Aldol
HO O O
coumarin SCoA O OH OH
(umbelliferone, HO stilbene
HO HO O SCoA HO O OH
Apiaceae)
(resveratrol, grape)
stilbenecarboxylic acid
PKS
n x malonyl-CoA
OH OH OH
SCoA
O O HO OH CHI HO O
R CHS
n
flavanone
HO O O chalcone (naringenin,
alkenylphenols OH O OH O
orange, tomato, etc)

Kato and Pezzuto (2012) Phytochemistry and Pharmacognosy,

Encyclopedia of Life Support Systems 1-14.
 Meroterpenoides
(merus (grego): parte, parcial ou fragmento

Geris and Simpson (2009) Nat. Prod. Rep., 26, 1063.

Cornforth, J. W. (1968) Chem. Br., 1968, 4, 102.

Prenylation reaction in
aromatic compounds
 O
Aminoacid
precursor OH
NH2
HO
L-tirosina

Alkaloids
RO
HO O

N
NH2 O O
HO
OMe NMe
OH
berberina
mescalina (Coptis japonica) HO morfina (R=H)
(peiote, OMe codeína (R=Me)
Lophophora williansii) (Papaver somniferum)
The biosynthesis of the BIAs.

Branch pathways are color-coded: white, simple BIAs; blue,

morphinan alkaloids; purple, laudanine; orange,
benzophenanthridine alkaloids; red, protoberberine alkaloids;
green, aporphine alkaloids.

Enzymes for which cognate cDNAs have been isolated are

highlighted in green, while red denotes those with
characterized activities.

Abbreviations:
BBE, berberine bridge enzyme; CAS, canadine synthase;
CFS, cheilanthifoline synthase; CNMT, coclaurine N-
methyltransferase; COR, codeinone reductase; CTS,
corytuberine synthase; DBOX, dihydrobenzophenanthridine
oxidase; DRR, 1,2-dehydroreticuline reductase; DRS, 1,2-
dehydroreticuline synthase; MSH, N-methylstylopine 14-
hydroxylase; NCS, norcoclaurine synthase; NMCH, N-
methylcoclaurine 3′-hydroxylase; 4′OMT, 3′-hydroxy-N-
methylcoclaurine 4′-O-methyltransferase; 6OMT,
norcoclaurine 6-O-methyltransferase; 7OMT, reticuline 7-O-
methyltransferase; PRH, protopine 6-hydroxylase; SalAT,
salutaridinol-7-O-acetyltransferase; SalR, salutaridine: NADPH
7-oxidoreductase; SalSyn, salutaridine synthase; SanR,
sanguinarine reductase; SOMT, scoulerine-9-O-
methyltransferase; STOX, (S)-tetrahydroprotoberberine
oxidase; STS, stylopine synthase; THS, thebaine synthase;
TNMT, tetrahydroprotoberberine cis-N-methyltransferase.

Liscombe and Facchini (2008 ) Current Opinion in Biotechnology, 19:173–180

Classical methods to study biosynthesis

Recognition of precursors

Test of hypothesis by in vivo feeding

labeled precursors

Study of sequence of reaction

and mechanism
 Produtos naturais como
Fonte para estudos de mecanismos
de reações orgânicas

108
 Biogenesis of natural products

Curcuma zedoaria (Zingiberaceae)

 Biogenesis of natural products

HO

HO COSCoA
CH3

HO

Yoshida et al. 2018. Phytochemistry Letters 25, 136-140.

Antileukemic activity (IC50 mM) of triketones from
Peperomia alata and P. trineura
OH O OH O
2 2 O
1' n 1' n
1 1
O 1a n=1 O O
6 6
1b n=7
2a n=1
2b n=13
1a (49.11/41.66) 2c n=15

1b (inactive/inactive) 2a (79.22/66.70)
2b (inactive/191.89)
2c (inactive/103.93)
OH O
16'
2
1'
7 10' 11'
1

6 O 3a R=H Vincristine 0.57

3b R=OH
R Gleevec 0.25

3a (-/-) K562/Nalm6
3b (26.04/14.29)

Ferreira et al., J. 2014. J. Nat. Prod. 77, 1377-1382.

 Occurrence of polyketides
OH O

O OH O
OH

RO OH

OH O

O OH O
OH

OH
Virola sebifera, V. elongata
(Myristicaceae) Kato et al., (1985) Phytochemistry, 24, 553.

OH O

HO OH

Zonaria species (Dyctiotaceae)

Gerwick W and Fenical W (1982) Phytochemistry, 21, 633.
Biosynthesis of 2-acylphloroglucinol type polyketides
O

CoAS

–
PKS III O O

SCoA

O O

R OH O
O O
R
SCoA

O
OH O OH
R

OH

+
OH O

RO OH
 Biogenesis of natural products

OH HO
OMe OMe

OMe OMe
O O

O O
O O
Magnostelin A

Kato et al. 1990. Phytochemistry 29, 1799-1810.

Podophyllum hexandrum Forsythia oleraceae

Sesamun indicum Araucaria angustifolia

 Sesame lignans: an interesting
biosynthetic question !!!
(Norman Lewis)
O
O
O
O
O

??? O O

O
O
O
O O
sesamin O sesamolin

Sesamun indicum (Pedaliaceae)

Kato, M. J., Chu, A., Davin, L. B., Lewis, N. G., 1998.

Biosynthesis of antioxidant lignans in Sesamum
indicum seeds. Phytochemistry 47, 583-591.
 OMe OMe OMe
OH OH OH OH
O OH OH

2x
OMe O O HO
OH HO HO HO
OMe OMe OMe
coniferyl
alcohol (+)-pinoresinol (+)-lariciresinol (-)-secoisolariciresinol
OMe
OH
O Forsythia intermedia
Biosynthesis (+)-piperitol Katayama et al. 1992
Katayama et al. 1993
O
of O O
Dinkova-Kostova et al. 1997
O
lignans O
O

(+)-sesamin
O
O
O O
Sesamun indicum
O
O O
Kato et al . 1998.
(+)-sesamolin Jiao et al. (1998).
O
O
O
 PROTOFLAVANONES:
PRIMITIVE CHARACTER RETAINED OR CONVERGENCE
EVOLUTION?
OMe
OMe
H
MeO O

cytotoxic effects OH
H
OH

on ovarian cancer OH O MeO O

OH
MDAH-2774 and
SKOV3 cells O
OH O
Ongokea gore (Angueuk)
HO O Olacaceae (Santalales)
OH
Jerz et al., (2005) Phytochemistry 66, 169.
OH O

Protoapigenone Freitas et al., (2013)

HO O

MeO O
OH
MeO
OH O

O
HO
H3CO O
Piper carniconnectivum
Thelypteris torresiana
H3CO
8
'Eco Maiden Lace' OH O

Chang et al., (2008) 325, 841-849; Chang et al., Cancer Lett (2008) 267, 85-95.
 Peperomins
in Piperaceae O O
R2O R2O
O O
R1O R1O

OR4 OR4
OR3 OR3
dibenzylbutyrolactone
peperomins lignans

O OH Secolignans
R2O MeO
in Myristicaceae
RO MeO

OR4
OR3 O
O
aryltetralin lignans secotetralin lignan

Kato et al., (1990) Phytochemistry 26, 179.

Estudos de produtos naturais e
de sua biossíntese

Estruturas de proteínas e genes

envolvidos na biossíntese de
produtos naturais

Novas reações e novos

mecanismos de reações
orgânicas
 “Science in the future should provide answer to questions
related to as the nature works, which is the essential condition
for the future of life in our planet“

O. R. Gottlieb
(Founder of phytochemistry in Brasil)
"Advanced Topics in Natural Products "
Massuo J. Kato (IQUSP)

Definitions
and theories on
evolutionary role

Economical
Chemical Natural Products importance
ecology throughout history

Methods for Bioprospecting

isolation and
analysis Biosynthetic
pathways