Development of Nutraceuticals targeting cancer

Catarina DuarteFood & Health Unit

Nutraceuticals and controlled Delivery LabIBET/ ITQB-UNLOeiras, Portugal

�Green Processes for developing health promoting products

Extraction /Fraccionation of Natural Compounds

Research interests:

Formulation – particle formation, encapsulation and

impregnation

�Characterization and pre-clinical Bioactivity evaluation of bioproducts

Chemical, enzymatic and cell-based assays

Food&Health Unit Nutraceuticals and Controlled Delivery Lab

� POH was detected in

10%EtOH and 20%EtOH

extracts

Serra et al. (2010) J Supercri. Fluids, 55, 184-191

Sweet cherriesDevelopment of POH-rich extracts

� Application of High Pressure Technology for the development of natural extracts

rich in perillyl alcohol (POH)

Raw material:

Sweet cherry culls

(Saco variety)

POH

96h

�Only two extracts (10%EtOH and 20%EtOH) inhibited cancer cell growth in a

time dependent manner and this effect is related with the presence of Perillyl

Alcohol

� Cherry extracts induced cell cycle arrest in a different cell cycle checkpoint

than doxorrubicin suggesting that they can be used in combination with the

drug in chemotherapy

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Antiproliferative effect on HT29 cells (human colorectal cancer cell line)• Cell cycle analysis

Controlo

10% EtOH

20% EtOH

• Cell viability after 24, 48, 72 and 96h of proliferation

Serra et al. (2010) J Supercrit. Fluids, 55, 184-191

Serra et al. (2011) J Supercrit. Fluids,55 1007-10131st prize: “Inovation and Develo+ment of Products and Services” category

Sweet cherriesDevelopment of POH-rich extracts

Minutes

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Detector 1-280nmLaranja (3) TS Ago2913076.dat

Detector 1-280nmLaranja (4) TS Ago2913077.dat

Detector 1-280nmLaranja Dil 1:10Jul2413094.dat

Sinensetin

Nobiletin

Tangeretin

� Process: CO2+ EtOH (80:20 v/v); P=25MPa; T=323K; t=60min.

Tangeretina

Nobiletin

Sinensetin

Inês Silva (2013), Tese de mestrado

Citrus peelsDevelopment of PMF-rich extracts

� Application of Supercritical Fluid Technology for the development of natural extracts

rich in polymetoxylated flavones (PMF)

� Sinensetin, tangeritin and nobiletin were

identified in citrus pell extracts.

Raw material:

Orange peels(by-products of fruit juice

production)

Induction of apoptosis on HT29 cell spheroids

Control Citrus extract

(Red: viable cells; Green: apoptotic cells)

Cell viability• 2D model

� Orange extract inhibited the proliferation of

HT29 cells (antiproliferative effect was

dependent on spheroid size).

� Orange extract induced apoptosis in HT29

spheroids(apoptotic cells are mainly located

on the surface of the spheroid).

• 3D model

Cell monolayer Cell spheroids with different sizes

Antiproliferative effect on HT29 cells (human colorectal cancer cell line)

Inês Silva (2013), Master thesis

Citrus peelsDevelopment of PMF-rich extracts

Opuntia ficus-indicaDevelopment of terpene-rich extracts

Raw material:

Opuntia ficus-indica(by-products of fruit juice production)

� Process: CO2+ EtOH+ H2O mixtures; P=20MPa; T=313K; t=180min.

� Application of Pressurized Liquid Extracton for the development

of natural extracts rich in terpenes

fried lup #1 #2 #3 #4 #5 #6 #7 #8 #9 #10 fried lup #1 #2 #3 #4 #5 #6 #7 #8 #9 #10

Terpene

detection by TLC

� Opuntia extracts obtained with mixtures of CO2 and

EtOH presented high terpene content

� Opuntia extract obtained with CO2:EtOH (60:40v/v) mixture (PLE2) presented the highest

antiproliferative effect. This effect is probably due to the presence of terpenes.

� PLE-2 extracts increased drug sensitivity on HT29 dx cell population cells .

Antiproliferative effect on HT29 cells (human colorectal cancer cell line)

• Cell viability after 24 h of proliferation

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Doxorubicin (µM)

Doxorubicin in HT29 dx

Doxorubicin in HT29

PLE-2 + Doxo in HT29 dx

• Chemosensitization effect in a drug resistant cell sub-

population (HT29 dx)

Joana Poejo(2014), Master thesis

Opuntia ficus-indicaDevelopment of terpene-rich extracts

e-mail: cduarte@itqb.unl.pt;

http://www.itqb.unl.pt/labs/nutraceuticals-and-delivery

www.ibet.pt

Laboratório de Nutracêuticos e

Libertação Controlada

Oeiras, Portugal