1.

Aspectos históricos, legais e

éticos. Desafios e Oportunidades
na Fitoterapia Médica

Prof. Dr. Niraldo Paulino

 Parte 3
APRESENTAÇÃO DO PRODUTO

Um exemplo super-interessante de
estudo científico de Fitoterápico

Uma abordagem completa do estudo

farmacocinético de um produto farmacêutico
derivado de planta medicinal em humanos.
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FERNBLOCK
 FERNBLOCK (EPL):
descrição botânica
• Polypodium leucotomos

• Familia Polypodiaceae
• Gênero Phebodium
– Crescem naturalmente como plantas epífitas em
diversas palmeiras

– América Central
• “Calaguala”
• Medicina popular
– Antiinflamatorio (psoriasis, dermatitis atópica)
 Reprodução

Por meio de esporos

– Que são lançados ao

vento e quando
encontram condições
favoráveis, germinan
dando lugar a novas
plantas.

Planta adulta de Phlebodium

Germinação controlada
 Rosetas e esporócitos sob germinação
controlada
.
Plantas desenvolvidas
 FERNBLOCK (EPL): cultivo

o Plantação localizada em no Lago YOJOA

HONDURAS em Olancho.
o Padronização das condições de cultivo
o Sem uso de defensivos agrícolas.
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FERNBLOCK
 FERNBLOCK (EPL):
composição
• Extrato aquoso constituido por:
– POLIFENÓIS: 3-10 % (p/p)
• Ácido Ferúlico
• Ácido Cafeico
• Ácido Cinámico

– MONOSACÁRIDEOS: 1-12 % (p/p)

• Fructose, manose e glicose

– FLAVONÓIDES

– EPL é una fonte natural de substâncias antioxidantes, principalmente

rico em derivados fenólicos
FERNBLOCK (EPL):
Laudo de análise

GARANTIA DE QUALIDADE

Reprodutibilidade na produção
do fitomedicamento!!
In summary, the presence of both free and conjugated phenolics, such as caffeic and chlorogenic acids,
and the presence of different chlorogenic isomers point toward the relevance of the shikimik pathway in
ferns, and the importance of phenolic compounds as secondary metabolites related to plant
photoprotection (28). In addition to photoisomerization of cinnamic and related phenolic acids, the in
vivo photocyclodimerization has been described and related to cell wall stabilization (29). The presence
of cyclodimers of the phenolic acids herein described, either in Fernblock or as a consequence of UV
irradiation of the extract, is currently under study in our laboratories.
3,4-Dihydroxybenzoic acid Chlorogenic acid

Vanillic acid Caffeic acid

4-Hydroxybenzoic acid
4-hydroxycinnamic acid
Neochlorogenic acid

Ferulic acid
 ATENÇÃO!!
“Nem tudo que se parece é igual...”
COMPARATIVE ANALYSIS BETWEEN IFC Polypodium leucotomos extract
(batch P-07) and VIAFARMA (HUZHOU) Polypodium leucotomos extract
(batch 20070418) (Internal code IFCE 1195):
IFC (PLE) VIAFARMA (PLE)
Advantages Disadvantages
1.In vivo photoprotector 1. No photoprotective activity in vivo
2.Natural water based extraction 2.Chemical extraction process
process
3.Phenol content and in vitro
photoprotective activity the same if not
better
4. Demonstrated anti-oxidant activity
5. Over 12 clinical trials In leading
Industrial Farmacéutica Cantabria, institution like Harvard and Sloane
Kettering demonstrating;
S.A.  Anti-oxidant activity

Pirita, 6 
Anti-aging activity
Photo-Inmuno Protection
28850 Torrejón de Ardoz  Photo protection
6. Over 15 years farmacovigilance and
MADRID (España) safety data.
7.Registered and commercialed in over
Tel.: (34) 91 676 90 82 42 countries
8. Registered with USFDA as ODI and
Fax: (34) 91 677 20 41 Self Affirmed GRAS
ifc@ifc.es 10. On-going pre-clinical and clinical
research.
11. Certified GLP/GMP extraction
processes assuring consistent quality
and stability.
12. Certified QA/QC processes
assuring consistent quality and
stability.
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In summary, these results underline the antioxidant and photoimmunoprotective properties of the PL extract
orally given in an in vivo model. PL-treated animals are less affected by UVR irradiation, chiefly noted in
decreased GSSG levels and a more favourable GSH⁄ GSSG ratio, and inhibition of UVR-mediated LC
depletion. These and other results strongly suggest the need to extend the concept of photoprotection,
currently restricted mainly to erythema development, which is the most extended criteria to evaluate the
efficacy of photoprotective measures. Oral administration of such antioxidant, photoprotective substances
would provide an excellent, hassle-free complement to the recommended use of topical sunscreens.
Overall, the effects of P. leucotomos predominated with or without exposure to UVA or UVB
radiation in both keratinocytes and fibroblasts, respectively. PL improved membrane integrity,
inhibited lipid peroxidation, stimulated elastin expression and inhibited MMP-1 expression in both
fibroblasts and keratinocytes. The only exception to the predominant effects of P. leucotomos was
in the inhibition of MMP-1 expression only in combination with UV radiation, particularly UVB, in
fibroblasts
REGULAÇÃO BIOQUIMICA NA CÉLULA

O sistema bioquímico
complexo que produz
espontaneamente
formas radicalares de
oxigênio reativo (ROS)
a cada movimento de
respiração celular.
Esses radicias livres
são consumidos pelos
nossos sistemas de
defesa antioxidante:
SOD, CAT, GSHpx, etc...
REGULAÇÃO DA RESPOSTA ANTIOXIDANTE

Além disso, fatores ambientais:

inflamação, radiação, agentes
químicos, lesão de reperfusão,
podem induzir radicais livres
adicionais nas células levando a
danos celulares.
 Efeito do P. leucotomos na reação oxidativa
induzida por radiação solar
P. leucotomos e seus derivados Polypodium leucotomos
fenólicos e flavonóides apresentam
forte atividade ANTIOXIDANTE
e SEQUESTRANTE DE
RADICAIS LIVRES

Geração de radicais livres

Radical Radical Radical Radical Outros

superóxido peroxil Alcoxil Hidroxil

P. leucotomos e seus derivados

fenólicos e flavonóides apresentam
forte atividade INIBITÓRIA Oxidação de proteínas
SOBRE OS FATORES DE
TRANSCRIÇÃO NUCLEAR
NFKapaB e AP1
Polypodium leucotomos

Controle da Controle da Mudança estrutural das proteínas

transcrição gênica diferenciação de regeneração celular
celular
In summary, P. leucotomos demonstrated dual protective eVects on the ECM for anti-skin aging via the inhibition
of the ECM proteolytic enzymes and the stimulation of TIMPs, structural ECM collagens (types I, III, V), and TGF- in
Wbroblasts. P. leucotomos’s anti-cancer eVects encompass the inhibition of MMPs and TGF- and stimulation of
TIMPs in melanoma cells. In addition, P. leucotomos anti-aging and anti-cancer mechanism lies in its direct
inhibition of MMPs. The eVects of P. leucotomos on Wbroblasts and melanoma cells may be partly via its cell-
speciWc regulation of TGF- expression and partly via its antioxidant property.
Polypodium leucotomos is used for the treatment of psoriasis (Difur®) and for photoprotection (Fernblock®) [18,
20]. Its topical or oral administration is tolerated without toxicity; for protection against UV radiation or PUVA
(Psoralen § UVA) mediated phototoxicity, depletion of langerhans cells, erythema and skin damage [18, 20, 21, 23,
35]. P. leucotomos is readily absorbed and metabolized cellularly [18]. We therefore infer an in vivo potential of P.
leucotomos for antiskin aging and anti-cancer, respectively, via speciWc beneWcial regulation of ECM remodeling.
 Efeito do P. leucotomos na reação inflamatória tópica
induzida por radiação solar
Estímulo inflamatório

Epiderme ERITEMA Epiderme

Carrageina

EDEMA
Vaso

AA PGs
COX

Apoptose
ou Necrose

Macrófagos Linfócitos PMN

Citocinas
NO PGE2 O2- ROS Inflamação
 Redução da inflamação e da angiogênese
Quimioterápicos, LPS
associada ao P. leucotomos
Estímulos
TLR
Ativação de
receptores tipo TLR
Myd-88 IRAK
Desencadeia uma
cascata de fosforilação
TRAF6

Ativação da NIK
quinase do IKK
IKK
IkappaB
Condições ideiais
IB para o crescimento
Fosforilação do
inibidor do NFkB  IL-12 do tumor e
p50 p65 metástase

Liberação do Fator  IL1

de Transcrição p50 p65 Inflamação:
Nuclear NFkB  TNF- aumento da
permeabilidade
Translocação  iNOS NO vascular,
vasodilatação,
nuclear para a
 COX-2 PG migração celular,
ativação da
etc…
expressão gênica
Expressão de
inúmeros genes Expressão de Expressão de Aumento da
pró-inflamatórios genes das citocinas genes da produção de PGE2,
IL1, 12 e TNF- cicloxigenase 2 PGF2 ...
 Photodermatology, Photoimmunology & Photomedicine 24, 134–141 2007

Specifically, PL has the capacity to neutralize free radicals, membrane-lipid peroxidation, and various ROS
(19, 20). It is possible that PL’s ability to prevent UVB radiation-induced immunosuppression and skin
ancer in mice is accomplished through its antioxidant properties as other antioxidants have been shown to
inhibit UVB-induced immune suppression in mouse models (21–24). ROS are thought to be involved in the
in the decrease in Langerhans cell numbers and the inhibition of CHS observed after exposure of murine
skin to UVB radiation (21) and ROS-induced lipid peroxidation of membrane lipids may impair antigen
presenting function in the remaining Langerhans cell. In addition, ROS increase TNF-a production (25), a
mediator important in CHS suppression in UVB-irradiated mouse skin (26). In this regard, PL has been
shown to inhibit TNF-a release stimulated by exposure of human keratinocytes to solar simulator radiation
(27). Furthermore, PL inhibitsthe photoisomerization of trans-urocanic acid to cis-urocanic acid, another mediator
implicated in UVB-induced immune suppression (28).
In conclusion, this study shows that anapsos is able to modulate the immune
response after surgery, by expressing an inhibitory effect on Th2 pathway
activation with no effect on Th1 pathway activation. This might be very important
in trauma and clinical surgery with regard to modulator treatment of the
postoperative immune response, since anapsos might prevent the shifting
Th1/Th2 balance and, in addition, preserve inflammationfollowing tissue repair
processes, and avoid “hyperinflammation” consecutive to immunosuppression
induced by surgery.
Mecanismos imunomodulador que justifica a melhoria da
resposta imune após o uso de P. leucotomos
Estímulos carcinogenicos
Ativação da p38-MAPK Carcinogenic externos
stimulus
Carcinogenic Ativação de proteinas da
stimulus classe Ras (GTPase)
Rac2 Cdc42
TLR
GTP Prenilação pós-
P. leucotomos translacional
Rac2 Cdc42
Myd-88 IRAK GDP
+
P Ativação de proteinas
TRAF6 MKK3 quinases ativadas por
p38 MKK6 mitógenos
NIK
IKK Ativação do fator de Aumento da
IKK modulação imunologica resposta
imune celular

IB
 IL-12
p50 p65
 IL-1
p50 p65
 TNF-
P. leucotomos
P. leucotomos
 iNOS NO

 COX-2 PG
In addition, PL downregulates TNF-a and iNOS expression induced by SSR. UV radiation induces TNF-a
expression (42). Interestingly, UV irradiation also induces expression of TNF-a receptors (43), which
suggests that TNF-a mediates the inflammatory reaction in skin induced by UV irradiation. On the other
hand, iNOS is also induced by UV irradiation in HaCaT cells (44). Increased iNOS expression causes
elevated levels of NO, which is one of the causal factors of erythema formation. The inhibitory effect of PL
on NO production and iNOS expression strongly suggests that NO increase is caused by de novo
synthesized iNOS. Finally, TNF-a and iNOS up-regulation are linked to increased apoptosis [reviewed in
Refs. (45,46)], thus inhibition of UV-mediated TNF-a and iNOS expression by PL is a likely cause for the
observed decrease in apoptosis. In summary, these findings greatly extend previous observations on the
photoprotective effect of PL, and represent a rigorous approach to explain its molecular basis, providing a
rationale for increased cell survival, decreased inflammation and changes in pigmentation observed both in
vivo and in vitro.
Conclusions
Originally described as a natural alternative for the treatment of inflammatory skin disorders such as psoriasis, P. leucotomos is a
more powerful tool than previously thought. It combines extremely low toxicity with proven beneficial effects, even with oral
administration. These include antioxidant properties, the capability to inhibit t-UCA photoisomerization (which makes it a very
valuable additive in sunscreen formulations), and the ability to block UV-induced apoptosis and DNA photodamage
andimmunosuppression. It seems clear that P. leucotomos is not an alternative to traditional sunscreen use, but it provides a very
effective complement for sensitive skin phototypes and adds extra protec- S. Gonzalez, J.L. Alonso-Lebrero, R. Del Rio and P.
Jaen 7 Table II: Beneficial effects of P. leucotomos. On... Molecular effects Short-term effects Long-term effects References
Generation of free Inhibits ROS Relieves photoaging, (32, 62, 75) radicals generation prevents tumor formation Generation of
other Inhibits t-UCA (Predicted) (64) deleterious photoisomerization, Relieves immunometabolites photodecomposition
suppression and tumor formation Inflammation Inhibits cell death and Reduces erythema, Relieves photoaging, (35, 45, 50, 51,
53) apoptosis sunburn* prevents tumor formation DNA damage Decreases the Reduces SOS (Predicted) Inhibits (53) formation of
response tumorigenesis pyrimidine dimers Immunosuppression Prevents depletion (Predicted) Reduces (Predicted) Inhibits (51,
53, 73, 74) of eLC recurrent infections chronic inflammation and cancer *Oral administration of P. leucotomos (1,080 mg)
increases skin tolerance to sunlight by a factor of 2.81 (48). ROS = reactive oxygen species; UCA = urocanic acid; SOS = the
SOS response: the depression of multiple genes encoding DNA repair proteins, leading to more rapid DNA repair and enhanced
bacterial survival; eLC = epidermal Langerhans cells. tion in cases in which exposure to UV radiation cannot be avoided, such as
those in UVB phototherapy and psoralens-UVA treatments.
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Recentes trabalhos científicos (2012) sobre P. leucotomos