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Olive Polyphenols and Malaria, a back-to-the-future Antiprotozoal strategy

  • Fulbright Program


Recently Olive polyphenols are being considered for their potential in the treatment of malaria, however medical literature’s historical reports indicate that reports about the benefits of olive extracts in the treatment of malaria date as far back as 1800. In this draft I discuss evidences validating the hypothesis of a potential role for olive polyphenols in an antiprotozoal strategies for the treatment of Malaria via the findings relating to a commercially available proprietary formulation of olive polyphenols.
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Olive Polyphenols and Malaria, a back-to-the-future Antiprotozoal strategy
©Paolo Pontoniere
Malaria is an infectious disease caused by parasitic protozoans belonging to the genus
Plasmodium.1 Mosquito-borne, the condition is generally transmitted by an infected Anopheles
female. Biting the host the mosquito introduces the parasites—contained in its saliva2--into the
carrier--a person or other warm-blooded animal. The symptoms usually develop ten to fifteen
days after being bitten. Carried to the liver sporozoite forms of the parasite, injected under the
skin by the mosquito bite, evolve into the merozoite form.3 The invasion of hepatocytes is
mediated by parasite surface protein like CSP. Subsequent infection of red blood cells (RBCs) by
merozoites causes malaria disease via aberrant cytokine production and sequestration of parasite-
infected red blood cells (pRBCs) to host endothelium. Cerebral malaria resulting from
microvasculature sequestration in the brain can result in life-lasting neurological impairment or
death.4 There are approximately 225600 million new malaria infections worldwide annually, with
severe and cerebral malaria representing significant causes of death internationally. Prevention
through mosquito control measures such as using such mosquitoes nets and insect repellents, or
spraying insecticides and draining standing water can help to reduce the diffusion of the disease
The treatment of choice for malaria is a cocktail of medications including artemisinin,
mefloquine, lumefantrine, or sulfadoxine/pyrimethamine2,5.
Quinine along with doxycycline is used when artemisinin is not available. Occasionally a
prescription of sulfadoxine/pyrimethamine is recommended in infants and pregnant women in
their first trimester of pregnancy in areas where rates of malaria are very high.6
Despite a dramatic need for a safe-effective-and long-lasting solution to the disease, and ongoing
efforts to produce a viable vaccine, as of today no final remedy has been found, although efforts
1 Cowman AF, Berry D, Baum J. The cellular and molecular basis for malaria parasite invasion of the human red
blood cell. The Journal of Cell Biology. 2012;198(6):961-971. doi:10.1083/jcb.201206112.
2 Malaria Fact sheet N°94. WHO. March 2014.
3 Lucy Megumi Yamauchi, Alida Coppi, Georges Snounou and Photini Sinnis, Plasmodium sporozoites trickle out
of the injection site., Cell Microbiol. 2007 May 1; 9(5): 1215–1222.
4 Idro R, Marsh K, John CC, Newton CR. Cerebral Malaria; Mechanisms Of Brain Injury And Strategies For
Improved Neuro-Cognitive Outcome. Pediatric research. 2010;68(4):267-274.
55Nosten F, White NJ. Artemisinin-based combination treatment of falciparum malaria. Am J Trop Med Hyg. 2007
Dec;77(6 Suppl):181-92.
6 Peters PJ1, Thigpen MC, Parise ME, Newman RD. Safety and toxicity of sulfadoxine/pyrimethamine:
implications for malaria prevention in pregnancy using intermittent preventive treatment. Drug Saf. 2007;30(6):481-
to develop one are ongoing.7 Furthermore, resistance among the parasites has expanded to
several antimalarial medications; for example, chloroquine-resistant P. falciparum has spread to
most malarial areas, and resistance to artemisinin has become a problem in some parts of
Southeast Asia.8
The disease is widespread in tropical and subtropical regions around the equator, including Sub-
Sahara Africa, Asia, and Latin America.
Malaria is associated with poverty and has a significant adverse effect on economic
development.9 It is estimated that in Africa malaria-induced increased in healthcare costs amount
to $12 billion year due to loss of work hours, and adverse effects on tourism.10 The World Health
Organization reports there were 198 million cases of malaria worldwide in 2013.11,12 This
resulted in an estimated 584,000 to 855,000 deaths, the majority of which--90%--occurred in
Olive polyphenols and Malaria, it’s back to the future
Recently Olive polyphenols are being considered for their potential in the treatment of malaria,
however medical literature’s historical reports indicate that expounding about the benefits of
olive extracts in the treatment of malaria date as far back as 1800.
In 1854, Hanbury published an article in the Pharmaceutical Journal of Provincial
Transactions relating that a "decoction of the leaves" of the olive tree had been
found to be extremely effective in reducing fevers due to a severe, and otherwise
often fatal, a disease that had swept the island of Mytilene in 18431. The olive
leaf extract was reported subsequently to be more effective in its fever-lowering
properties than quinine. Hanbury recalled that similar observations had been made
in France and Spain many years previously (between 1811 and 1828). It appears
that, in the early 19th century, Spanish physicians sometimes prescribed olive
leaves as a "febrifuge," and consequently, during the Spanish war of 1808—1813,
the French Officers de Sante often used them to treat cases of "intermittent fever."
Hanbury went on the describe how Pallas, following observations of clinical
benefits made an analysis of the leaves and young bark of the olive tree and found
them to contain, among other compounds, a bitter crystallizable substance which
he designated as "Jacqueline." Pallas ascribed most of the "febrifuge" properties
of the olive tree to Vauqueline.13
7 Benjamin Mordmüller, Güzin Surat […] Peter G. Kremsner. Sterile protection against human malaria by
chemoattenuated PfSPZ vaccine. Nature 542, 445–449 (23 February 2017)
8 Cui L, Mharakurwa S, Ndiaye D, Rathod PK, Rosenthal PJ. Antimalarial Drug Resistance: Literature Review and
Activities and Findings of the ICEMR Network. The American Journal of Tropical Medicine and Hygiene.
2015;93(3 Suppl):57-68. doi:10.4269/ajtmh.15-0007.
9 Behrens N. B., "Malaria: An update for physicians". Infectious Disease Clinics of North America. (2012). 26 (2): 243–
10 "Malaria Fact sheet N°94". WHO Retrieved February 4, 2019.
11. World Health Organization World Malaria Report 2014, WHO Geneva (2014), pp. 32–42. ISBN 978-92-4-156483-0
12 Gollin D, Zimmermann C Malaria: Disease Impacts and Long-Run Income Differences (August 2007). Institute for
the Study of Labor.
13 Amanda Jackson-Russell, Olive Leaf Extract , Positive Health issue 21 - August 1997
In 1906, another report stated that olive leaves were, in fact, superior to quinine for malarial
infections. Quinine was preferred, however, because it was easier to administer.
In studies performed by the Upjohn Company during the sixties with olive pumice, it was found
that calcium elenolate, a substance within Oleuropein, was effective against the malaria protozoa.
Preliminary reports from Latin America are promising. A full-fledged case of malaria at a clinic
in Mexico was cured with a dosage schedule of two olive leaf extract supplements every six
hours. A clinic report said that the 34-year-old female patient made a steady recovery and after
six months, “she was without any of the malaria symptoms, not even anemia or shivers. Her
breath is good. Her state of mind excellent and she does not show any signs of chronic or
contagious disease.”14
Nitric Oxide role
Recently, researchers, attention has been focusing on the role of nitric oxide (NO), and host
response in cerebral malaria continues to be elucidated, along with numerous known functions
relating to the cytokine, endovascular and cellular responses to infection with Plasmodium
falciparum. Evidence from diverse modes of inquiry suggests that NO may be critical to
modulate the immune response and to promote survival in patients with cerebral malaria.15
Furthermore working in a mouse model, Visioli et al. found that in the presence of endotoxin,
olive polyphenols (Oleuropein) increase production of nitric oxide with consequently increased
mobilization of macrophages.16 As recent as 2012 Bergmark et al. have proposed the use of
inhaled nitric oxide in cerebral malaria as a strategy to buy time to allow pharmacotherapy to be
effective. This line of investigation has culminated in the approval of 2 phase II randomized
prospective clinical trials in Uganda studying the use of inhaled NO as adjuvant therapy in
children with severe malaria. The strategy underlying both trials is to use the systemic anti-
inflammatory properties of inhaled NO to "buy time" for antiparasitic chemical therapy to lower
the parasite load.11
Protease inhibition and Maslinic acid
Anti and pro enzymatic activities of olive polyphenols are also being considered in the fight
against Malaria. In a 2009 University of Granada study, Luis M. de Pablo et al. found that
Maslinic acid causes protease inhibition in the invading protozoa – protease is necessary to the
parasite to invade the cell, thus indicating a potential use of olive polyphenols to reduce the
invasiveness of the protozoa.17
14 James R. Privitera, Olive Leaf Extract A New/Old Healing Bonanza for Mankind, Website:, Accessed 12/26/17 20:40
15 Bergmark, Brian MD, Bergmark, Regan MD, Beaudrap, Pierre De MD, PhD, Boum, Yap, Mwanga-
Amumpaire, Juliet MD; Carroll, Ryan MD, MPH, Zapol, Warren MD. Inhaled Nitric Oxide and Cerebral Malaria:
Basis of a Strategy for Buying Time for Pharmacotherapy. The Pediatric Infectious Disease Journal: December
2012 - Volume 31 - Issue 12 - p e250–e254doi: 10.1097/INF.0b013e318266c113
16 Visioli et al., Oleuropein, the Bitter Principle of Olives, Enhances nitric oxide production by mouse
Furthermore In vitro experiments using erythrocytes infected with Plasmodium falciparum
demonstrated that Maslinic acid (0.1−200 µM) inhibited the growth of the parasite in a dose-
dependent manner.18 At a concentration of 30 µM (close to the IC50), the triterpene reduced
parasitaemia to 4% (compared to 8% in untreated red blood cells) and slowed down the cell
cycle, since only the infective (schizonts) and immature (new rings) forms, but not the mature
forms (trophozoites), were observed in the erythrocytes. However, the removal of maslinic acid
from the medium permitted the infection to resume, meaning that the triterpene acts as a
parasitostatic agent. 19
HT and Inhibition of SREBP-1c/FAS pathway
A rationale for the use of Olive polyphenols in the treatment of malaria has also been proposed
because of the action that they exert on the SREBP-1c/FAS pathway for the synthesis of fatty
acids. The exponential rate at which malaria parasites multiply during asexual blood-stage
replication, the production of sporozoite in the midgut, and intrahepatic development of
Plasmodium-malaria parasites--which precedes the pathogenic intraerythrocytic phase--create
an essential need for building-block nutrients, including fatty acids that likely are necessary for
the parasite’s membrane formation.20 Plasmodium parasites obtain fatty acids either by
scavenging from the vertebrate host and mosquito vector or by producing fatty acids de novo via
the type 2 fatty acid biosynthesis pathway (FAS-II).21
In the direction of OP relevance to this application, a study on diet-induced metabolic syndrome
has shown that HT can prevent high-fat-diet (HFD)-induced obesity, hyperglycemia,
hyperlipidemia, and insulin resistance in C57BL/6 J mice after 17 weeks supplementation.
Within liver and skeletal muscle tissues, HT decreased HFD-induced lipid deposits through
inhibition of the SREBP-1c/FAS pathway and ameliorated HFD-induced oxidative stress by
enhancing antioxidant enzyme activities.22
17 Luis M. De Pablos, Gloria Gonza´lez, Raquel Rodrigues Andre´s Garcı´a Granados, Andre´s Parra,Antonio
Osuna. Action of a Pentacyclic Triterpenoid, Maslinic Acid, against Toxoplasma gondi. Biochemical and Molecular
Parasitology Group, Biotechnology Institute, Campus de FuentenueVa, UniVersity of Granada, 18071, Granada,
18 Carlos Moneriz, Patricia Marín-García, Andrés García-Granados, José M Bautista, Amalia Diez and Antonio
Puyet. Parasitostatic effect of maslinic acid. I. Growth arrest of Plasmodium falciparum intraerythrocytic stages.
Malaria Journal 2011, 10:82
19 Glòria Lozano-Mena , Marta Sánchez-González, M. Emília Juan, Joana M. Planas. Maslinic Acid, a Natural
Phytoalexin-Type Triterpene from Olives — A Promising Nutraceutical?, Molecules 2014, 19, 11538-11559
20 Ben C. L. van Schaijk, T. R. Santha Kumar, Martijn W. Vos, Adam Richman, Geert-Jan van Gemert, Tao Li,
Abraham G. Eappen, Kim C. Williamson, Belinda J. Morahan, Matt Fishbaugher, Mark Kennedy, Nelly Camargo,
Shahid M. Khan, Chris J. Janse, Kim Lee Sim Stephen L. Hoffman Stefan H. I. Kappe, Robert W. Sauerwein David
A. Fidock, andAshley M. Vaughan. Type II Fatty Acid Biosynthesis Is Essential for Plasmodium
falciparum Sporozoite Development in the Midgut of Anopheles Mosquitoes. Eukaryot Cell. 2014 May; 13(5): 550–
21 Sean B. Joseph, Bryan A. Laffitte, Parthive H. Patel, Michael A. Watson, Karen E. Matsukuma, Robert Walczak,
Jon L. Collins, Timothy F. Osborne, and Peter Tontonoz. Direct and Indirect Mechanisms for Regulation of Fatty
Acid Synthase Gene Expression by Liver X Receptors. THE JOURNAL OF BIOLOGICAL CHEMISTRY Vol. 277,
No. 13, Issue of March 29, pp. 11019–11025, 2002
22 Cao, K., Xu, J., Zou, X., Li, Y., Chen, C., Zheng, A. Feng, Z. (2014). Hydroxytyrosol prevents diet-induced
metabolic syndrome and attenuates mitochondrial abnormalities in obese mice. Free Radical Biology and Medicine,
Opportunistic Infective Agents
The role of opportunistic secondary infective agents in increased morbidity, mortality and
recurrence of Malaria has also been investigated. In this direction, Aflatoxin has emerged as one
the main culprits. Aflatoxin and malaria infection are tied by what appears to be a positive
feedback loop. The increase of one’s secretion (aflatoxin) increases host susceptibility to Malaria
infection while Malaria’s increased virulence renders the host more susceptible to Aflatoxin
toxicity. A study found that Aflatoxin-albumin adduct levels were higher in children who were
HBsAg (Hepatitis B surface’s Australia antigen) positive and in children with Plasmodium
falciparum parasitemia than in controls23
Children are particularly affected by aflatoxin exposure, which leads to stunted growth, delayed
development, liver damage, and liver cancer. Adults have a higher tolerance for exposure but are
also at risk. No animal species is immune.
Aflatoxins are among the most carcinogenic substances known. After entering the body,
aflatoxins may be metabolized by the liver to a reactive epoxide intermediate or hydroxylated to
become the less harmful aflatoxin M1. AFL-induced immune suppression and increased malaria
prevalence and severity of illness in endemic areas. In 1992 the International Journal of
Molecular Sciences reported that in the presence of 6 mg/mL oleuropein, production of aflatoxin
could be significantly decreased.24
Previous studies have indicated the inositol monophosphate (IMP) is a component of the malaria
parasite toxin that induces cytokines such as tumor necrosis factor (TNF).25-26It is a small peptide
of 18 or 19 amino acid residues and triggers intestinal secretion by elevating the intracellular
levels of cyclic guanosine 3'5' monophosphate (cGMP) Inhibition of STa-induced fluid
accumulation in suckling mouse by Ca 2÷ antagonists [Cit. Needed] and prostaglandin synthesis
blockers [Cit. Required] indicate that STa-mediated secretory response is not a direct process but
follows a cascade mechanism. Previous reports from our laboratory have also shown that, apart
from cGMP, two other second messengers, i.e., inositol triphosphate (IP 3) [Cit. Needed] and
diacylglycerol (DG) [Cit. needed] are derived from the receptor-mediated inositol phospholipid
breakdown initiated by the activation of phospholipase C (PLC) [Cit. Required] by STa. It is now
well recognized in many cellular systems that IP 3 mobilizes Ca 2÷ from some intracellular
stores [Cit. Needed] and DG activates the protein kinase C (PKC) either independently of or in
concert with the rise of cytosolic Ca 2 + [Cit. Needed]. Following STa stimulation the activity of
membrane-bound PKC has been found to be increased in rat enterocytes [Cit. Needed] and T84
67, 396–407.
23 Allen SJ , Wild CP, Wheeler JG, Riley EM, Montesano R, Bennett S, Whittle HC, Hall AJ, Greenwood BM.
Aflatoxin exposure, malaria and hepatitis B infection in rural Gambian
children. Trans R Soc Trop Med Hyg. 1992 Jul-Aug;86(4):426-30
24 Gourama H., Bullerman L.B. Effects of oleuropein on growth and aflatoxin production by Aspergillus
parasiticus. Lebensm. Wiss. Technol. 1987;20:226–228.
25 Kristensen G1, Jakobsen PH,. Plasmodium falciparum: characterization of toxin-associated proteins and
identification of a hemoglobin containing parasite cytokine stimulator. Exp Parasitol. 1996 Mar;82(2):147-54.]
26 Heat stable enterotoxin (STa) produced by enterotoxlgenic Escherichia coli causes secretory diarrhea in humans
and animals
Falciparum malaria can cause immune suppression sufficient to allow opportunistic infection
during the recovery phase. A patient is described to have died from a disseminated disease
with Aspergillus flavus and Absidia corymbifera, unresponsive to treatment with amphotericin
and voriconazole.28
The foodborne pathogen Staphylococcus aureus produces the virulent staphylococcal enterotoxin
A (SEA), a single chain protein which consists of 233 amino acid residues with a molecular
weight of 27078 Da. SEA is a
Superantigen that is reported to contribute to an animal (mastitis) and human (emesis, diarrhea,
atopic dermatitis, arthritis, and toxic shock) syndromes. Changes in the native structural integrity
may inactivate the toxin by preventing molecular interaction with cell membrane receptor sites
of their host cells. In the present study, we evaluated the ability of the pure olive compound 4-
hydroxytyrosol and a commercial olive powder called Hidrox-12, prepared by freeze-drying
olive juice, to inhibit S. aureus bacteria and SEA’s biological activity. Dilutions of both test
substances inactivated the pathogens. Two independent cell assays (BrdU incorporation into
newly synthesized DNA and glycyl-phenylalanyl-aminofluorocoumarin proteolysis)
demonstrated that the olive compound 4-hydroxytyrosol also inactivated the biological activity
of SEA at concentrations that were not toxic to the spleen cells. However, efforts to determine
inhibition of the toxin by Hidrox-12 were not successful because the olive powder was cytotoxic
to the spleen cells at concentrations found to be effective against the bacteria. The results suggest
that food-compatible and safe antitoxin olive compounds can be used to inactivate both
pathogens and toxins produced by the pathogens.29
In conclusion why Hidrox?
Because of its proven broad-spectrum anti-inflammatory and antioxidant properties (reduction of
TNF-α secretion, NfKb mobilization, Arachidonic acid mobilization, reduction of CRP, ROS
species, iNOS, COX2, oxidase enzymes, and tyrosine phosphorylation), of its small molecular
size, which allows the compound the cross the blood-brain-barrier of the host. Furthermore, in
virtue of its manufacturing process, Hidrox is the only formulation of olive polyphenols which
increases the expression of polyphenols in formulation while preserving the integrity of the
natural biological matrix in which the phenols have been generated. Henceexpressing many of
27 Alok Ghosh Chaudhuri, Uma Ganguly, Evidence for stimulation of the inositol triphosphate-Ca 2+ signaling
system in rat enterocytes by heat stable enterotoxin of Escherichia coli, Biochimica et Biophysica Acta 1267 (1995)
131-133, National Institute of Cholera and Enteric Diseases, P-33, C.LT. Road, Scheme XM, Beliaghata, Calcutta
700 010, India
28 [Desseminated Fungal Infection Following Falciparum Malaria, A.P.R. Wilson, S. Wright, G. Bellingan,Journal
of Infection Volume 40, Issue 2, March 2000, Pages 202–204]
29 Mendel Friedman, Reuven Rasooly, Paula M. Do, and Philip R. Henika, The Olive Compound 4-
Hydroxytyrosol Inactivates Staphylococcus aureus Bacteria and Staphylococcal Enterotoxin A (SEA) , Journal of
Food Science _ Vol. 76, Nr. 8, 2011
the qualities olive’s bioactive that have been found to be effective against Plasmodium invasion
of host’s erythrocytes, multiplication and in general in limiting morbidity and mortality
associated with Malaria.
Some relevant supporting studies for the role of Hidrox in Malaria
Hidrox reduces TNF-alpha1 expression in vivo and in vitro (Bitler et al. Journal
Nutrition, 2007 -CreAgri)
Hydroxytyrosol inhibits pro-inflammatory cytokines, iNOS, and COX-2 expression in
human monocytic cells ( Xiaomei Zhang et al., Naunyn-Schmied Arch Pharmacol (2009)
Hydroxytyrosol promotes mitochondrial biogenesis and mitochondrial function in 3T3-
L1 adipocytes (Jiejie Hao et al., Journal of Nutritional Biochemistry (2009)
Hidrox impairs cytokine and chemokine production in macrophages. (Richard N. et al.
Planta Med. 2011 Nov;77(17):1890-7. Pub 2011 Aug 9. DSM Nutritional Products)
HIDROX ® inhibited production of NO and PGE reflecting strong anti-inflammatory
activity. HIDROX ® and OVW diminished secretion of cytokines (IL-1 α, IL-1 β, IL-6,
IL-12, TNF- α), and chemokines (CXCL10/IP-10, CCL2/MCP-1). HT and OVW
concentration-dependently reduced the expression of genes of inducible nitric oxide
synthase (iNOS), IL-1 α, CXCL10/IP-10, MIP-1 β, matrix metalloproteinase-9, and
prostaglandin E synthase (PGES). The effects of HT were partly mediated VIA the NF-
κB pathway.
Hydroxytyrosol inhibits growth and cell proliferation and promotes high expression of
sfrp4 in rat mammary tumors. Granados-Principal et al. University of Granada, Granada,
Spain. Mol Nutr Food Res. 2011 May;55 Suppl 1:S117-26.
Hidrox may be an ideal, natural, non-toxic, inexpensive and sustainable, and easily
deployable option in the war against Plasmodium falciparum and the symptomatology
associated with malaria’s infection, and warrants further investigation.
The Many Faces of Olive Polyphenols 30
30 P.Pontoniere, D. Martiradonna, Inflammation and olive polyphenols :A perspective review of supporting
literature, AgroFOOD industry hi-tech - September/October 2012 - vol 23 n 5
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