Antimalarials Flashcards

1
Q

LO

A

identify the key events in the life cycle of the malaria parasite and how these may be affected by antimalarial agents

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
2
Q

Topics covered

A
  • The prevalence/ vector / parasite / disease / diagnosis/ susceptibility
  • Parasite life cycle
  • Vector control (including larvicides, insecticides, genetics)
  • Vaccine
  • Drugs for prophylaxis, acute treatment, and radical cure
  • Resistance and future therapies/targets
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
3
Q

What is malaria?

A

RBC are invaded by the Plasmodium species which is carried by mosquitoes

Its eukaryotic, parasitic protozoan

Infection by mosquito vector (and mother-foetus)

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
4
Q

Where is the term ‘malaria’ from and what is also sometimes referred to as?

A

Literally bad air (from Old Italian “mal” ‘aria”)

Sometimes known as “intermittent fever” / “marsh fever”

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
5
Q

When were cases of Malaria first recorded?

A

Recorded as far back as 3000 BC in China, India, and Egypt / Roman times in the UK (parasite in humans ~100,000 years)

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
6
Q

Why do the number of UK cases rise?

A

Number of UK cases on the rise due to global travel (~2,000 a year – ‘suitcase’ malaria)

Also climate change

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
7
Q

Malaria is described as being one of the ‘big three’ diseases, alongside what two others?

A

HIV

TB

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
8
Q

in 2019, what were the reported cases and deaths of malaria by the WHO

Who was it more prevelant in and why?

A

229 million cases a year / 409,000 deaths (WHO 2019)

Found mostly children <5 because they have not been exposed to it and have no defences against it

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
9
Q

What are the regions in which malaria is endemic?

endemic: when a diseases is generally found among certain populations and places

A

Primarily in tropical and subtropical regions as that is where the mosquitoes are present

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
10
Q

Tell me about the stats for the Morbidity and mortality in sub-saharan Africa where its mainly concentrated

A

Morbidity: The state of having a specific illness or condition

Mortality: Refers to the nunber of deaths that have occurred due to a specific illness or condition

Under 5s account for 52% of deaths (2/3rds for under 14s)

Sub-Saharan Africa: 94% cases and 95% of deaths

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
11
Q

The global cases are falling but, successes in this since the 2000 are slowing due to what factors?

A

‘Success’ – sustainability/removal of funds

Resistance (mosquito and Plasmodium)

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
12
Q

How is malaria transmitted?

A

Transmitted through the “bite” of the female mosquito genus Anopheles (~40 species associated with malaria)

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
13
Q

What happens when a mosquito bites its target?

A

Parasite initially ingested during a blood meal of an infected individual, it grows replicates in gut, and is then passed on during subsequent meal (8-15 sporozoites)

Sporozoites: a motile spore-like stage in the life cycle of some parasitic sporozoans (e.g. the malaria organism), that is typically the infective agent introduced into a host.

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
14
Q

Where are mosquitos usually found?

A

Abundant and distributed all over the globe, including the Arctic, but particularly widespread in tropical / sub-tropical regions, particularly around stagnant water (as larvae grow in stagnant water. Issue in places like Africa who will have large rainfalls and large water pools forming so trying to target this is quite difficult)

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
15
Q

How can the parasites be transmitted/

A

Parasites can be transmitted through blood transfusions and contaminated needles (but generally rare), as well as intra-uterine (mother-baby); can lead to complications and potentially miscarriage

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
16
Q

What are the 5 species of parasite that can form malaria and rank them from most to least dangerous

Which ones require radical cure and why?

A

P. falciparum (most serious / 90% deaths)

P. vivax*, ovale*, malariae (milder disease)

P. knowlesi (zoonotic - monkey & human)

*Requires radical cure- because they have a dormant state in the liver which can hide from the immune system and then erupt and come back later in life

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
17
Q

How does the parasite uptake the nutrients it requires from the red blood cells?

A

Uptake material from RBC; haemoglobin and protein component. Has a food vacuole

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
18
Q

Explain abit about the incubation period of the different malarial parasites

A

Can live in system for a few weeks without any symptoms. Only when its present in your blood is its known that you are infected

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
19
Q

Tell me what the initial symptoms are for the malarial parasites

A
  • flu like
  • headache
  • Photophobia
  • muscle aches
  • nausea
  • vomiting
  • chills (10-15 mins)
  • Fells cold but is actually feverish
  • Jaundice

Jaundice as RBC breaking down and bilirubin being present

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
20
Q

Tell me about the periodicity of the malarial parasites

A

Goes through rounds of replications once in RBC leading to cycles of symptoms- periodicity

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
21
Q

Tell me about the onset of diseases the the severity and duration (hr) for malarial parasites

A

Can cross BBB, clump RBC and this can lead to blocking blood vessels and brain swelling which leads to death- if survive it can lead to chronic conditions

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
22
Q

With the malarial parasite if left untreated or not treated properly it can lead to relapses or recrudescences, explain this

A

If untreated by drugs, then it can come back during your lifetime

Recrudescence is when you haven’t fully cleared from blood, not completely killed, will come back

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
23
Q

Why can one get anaemia from malarial parasites?

A

Anaemia as breaking over blood cells and have the side effects from this

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
24
Q

The disease and the 5 malarial parasites

A
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
25
Q

Some individuals can have genetic advantages which can help protect them against malaria. State 3 of them

A
  • sickle cell disease
  • Glucose-6-phosphate dehydrogenase deficiency
  • Lack of Duffy antigens on RBC
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
26
Q

How can sickle cell disease provide some protection against malaria?

A

Sickle Cell Disease (HbS, β-thalassemia etc.)

Heterozygous then can get resistance to malaria but homozygous you can get the disease

The sickle cells have membranes, stretched by their unusual shape, that become porous and leak nutrients that the parasites need to survive and the faulty cells eventually get eliminated quite fast by the organisms, destroying the parasite along the way.

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
27
Q

How can Glucose-6-phosphate dehydrogenase deficiency (X-linked) provide one protection against malaria?

A

Glucose-6-phosphate dehydrogenase deficiency (X-linked)

if lack enzyme then the RBC are more susceptible to redox damage and breakdown, malaria is more susceptible to redox damage so is less likely to infect these cells

As G6PD deficiency leads to increased oxidative stress in red blood cells, this may in turn have a negative influence on the parasite. As such, individuals who possess this mutation have some protection against malaria

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
28
Q

How can a lack of Duffy antigens provide one protection against malaria?

A

Lack of Duffy antigens on RBC

Duffy antigens plat a fundamental role on haematopoiesis (formation of blood cell components)

Duffy antigens are located on the surface of RBC

The protein encoded by this gene is a glycosylated membrane protein and a non-specific receptor for several chemokines

A lack of these antigens make the RBCs more resistant to invasion by a malarial parasite

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
29
Q

What may some people have that puts them more at risk to malaria?

A
  • Pregnancy
  • Co-infection: HIV, helminths
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
30
Q
A
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
31
Q

What are some techniques into the clinical diagnosis of malaria?

A

Microscopic examination of blood smears using Giemsa/Wright stain

Giemsa stain: specific to phosphate groups of DNA. Attaches to regions of DNA with high amounts of AT bonding

Wright stain: Facilitates the differentiation of blood cell types. classically a mixture of eosin (red) and methylene blue dyes

Antigen-capture assay (‘dipsticks’) - 100 parasites/µl lower limit of detection

PCR for strain detection (for determining drug resistance)

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
32
Q

Tell me the stages to the malarial life cycle and what can happen at each stage

A
  1. Injection of sporozoites into blood stream
  2. Sporozoites can pass quickly into the liver OR can remain dormant (Hypnozoites)
  3. Go through Asexual reproduction over roughly 10 days. Over this time no symptoms show
  4. Liver Schizonts are now present
  5. Merozoites are released into the blood where they can; invade RBC, asexually reproduce, cause cells to burst, repeat cycle, cause cyclic fever
  6. Blood Schizonts are now present
  7. Some Merozoites develop into Gametocytes that can circulate in the blood
  8. Moquito can ingest gametocytes, which develop into gametes and reproduce sexually
  9. Develop inot ookinetes that burrow through gut wall to form oocysts
  10. Oocyst bursts, releasing sporozoites that travel to salivary glands
  11. Cycle repeats when the mosquito bites another person
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
33
Q

What are sporozoites?

A

a motile spore-like stage in the life cycle of some parasitic sporozoans (e.g. the malaria organism), that is typically the infective agent introduced into a host.

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
34
Q

What are Hypnozoites?

A

Hypnozoites are dormant forms in the life cycles of certain parasitic protozoa that belong to the Phylum Apicomplexa (Sporozoa) and are best known for their probable association with latency and relapse in human malarial infections caused by Plasmodium ovale and P. vivax.

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
35
Q

What are Schizonts?

A

They are mature malarial parasites in host liver cells or red blood cells that are undergoing nuclear division by a process called schizogony

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
36
Q

What are gametocytes?

A

A eukaryotic germ cell that divides by mitosis into other gametocytes or by meiosis into gametides during gametogenesis

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
37
Q

What are ookinetes?

A

The motile zygote that forms when the microgamete (derived from the male gametocyte) fertilises the macrogamete (derived from the female gametocyte) during the sexual reproduction of certain sporozoans such as the malaria causing plasmodium

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
38
Q

What are oocysts?

A

A cyst containing a zygote formed by a parasitic protozoan such as the malaia parasite

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
39
Q

Whats schizogony?

A

Asexual reproduction by multiple fission, found in some protozoa

This occurs at a phenomenal rate

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
40
Q

What can Schizogony in parasites produce?

A

A single parasite of P. vivax can give rise to 250 million merozoites in 14 days!

To appreciate the action required of an antimalarial drug, note that the destruction of 94% of the parasites every 48 h will serve only to maintain equilibrium and not further reduce their numbers!- rapid reproduction

Produce a Schizont which can rapidly divide into many cells (merozoites) and infect

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
41
Q

What are some approaches being developed to eradicate the parasites (mainly P. falciparum as its the most deadly)

A
  • Vaccines to prevent infection
  • Drugs for radical cure
  • Drugs for prophylaxis (try to prevent infection. However these drugs cannot be taken over a long period of time due to severe side effects)
  • Drugs to treat acute attack
  • Drugs to prevent transmission
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
42
Q

When trying to control vectors of diseases, what are some of the things you could potentially target?

What does the combination of these approaches do?

A
  • Target the larvae life stage
  • Target adult moquito

The combination of these non-pharmacological approaches decreased the number of malaria cases in Africa by two thirds between 2000-15 (but stalling…)

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
43
Q

How do you target the larvae life stage?

A
  • Use of larvicides
  • Remove/ cover standing water e.g, drain swamps, oil films
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
44
Q

How can you target the adult mosquito?

A
  • Use of insecticides / repellents
  • Long-lasting insecticidal nets (LLINs)
  • Indoor residual spraying (IRS)
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
45
Q

Whats different forms can Larvicides be?

A
  • Chemical agents
  • Biological agents
  • Biodegradable oils
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
46
Q

Tell me about Larvicides as chemical agents and provide an example

A

Chemical agents e.g. Methoprene

Juvenile Hormone Analogue: increasing levels stalls larval development

Note yet routinely used in Sub-Saharan Africa (cost/easier to target adults/toxic to other aquatic life)

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
47
Q

Tell me about larvicides as biological agents

provide examples

A

Biological agents e.g., Bacillus thuringiensis var israelensis and Bacillus sphaericus

Apply pellets to water

Effective but complex considerations for deployment (mosquito species, water type etc.)

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
48
Q

What do biodegradable oils do as larvicides?

A

Prevent the larvae from breathing

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
49
Q

Give an example of a main insecticide

A

Dichlorodiphenyltrichlorethane (DDT)

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
50
Q

Tell me the following about the insecticide DDT…

  • when was it used
  • how does it act
A

Extensively used in 1950s and 1960s in US, Southern Europe and South America

Acts by opening voltage gated Na+ channels in neurons, causing them to fire spontaneously, which leads to spasms and eventual death (1000-fold higher affinity for insect channels = selective toxicity)

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
51
Q

Tell me about DDE bio-accumulates in the fat of animals

A

DDE bio-accumulates in the fat of animals (rarely excreted).

Responsible for the decline in many species of birds and fish and reproductive issues in humans (carcinogenic/endocrine disruptor) – largely banned in 1970s/80s

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
52
Q

Whats the Stockholm convention

A

The Stockholm Convention is a global treaty that aims to protect human health and the environment from the effects of persistent organic pollutants (POPs)

·

Stockholm Convention – should only be used for vector control (IRS)

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
53
Q

DDT…

A
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
54
Q

Give an example of another insecticide

A

Pyrethrins/ Pyrethroids

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
55
Q

Compare the used of Pyrethrins/ Pyrethroids as insecticides compared to DDT

A

Safer (Shorter half-life) but more expensive alternative to DTT

56
Q

What is the pyrethrin/ pyrethroid insecticide derived from?

A

Chrysanthemum sp. or synthetic

57
Q

Many DDT-pyrethrin resistant strains of mosquito have also evolved. Tell me the sort of mutations that have arised from them

A

upregulation of CYP450s GSTe2 genes – detoxify

Mutations to sodium channels

Sequestering proteins in mosquito legs!

58
Q

What can you combine pyrethrins with?

A

CYP450 inhibitors on LLINs

59
Q

Give some examples of insecticides that target acetylcholine

A

Organophosphates e.g., malathion

Carbamates e.g., Bendiocarb

Acetylcholine agonists at nAChRs

All three of these are Acetylcholine esterase inhibitors

60
Q

Tell me about Organophosphates e.g., malathion

A

Irreversible inhibition

Not typically used in residential settings – highly toxic

61
Q

Tell me about Carbamates e.g., Bendiocarb

A

Same mechanism but reversible

62
Q

Tell me about Acetylcholine agonists at nAChRs

A

Neonicotinoids: e.g., clothianidin - selective

General problems: environmental damage both to insects (pollinators – honeybees) and other animals

63
Q

Insecticides structures…

A
64
Q

Whats a technique used to control mosquitos?

A

The sterile insect technique (SIT)

65
Q

Tell me the stages to the sterile insect technique (SIT) for controling moquitos

A

and or

Wolbachia (bacterial) infection (Incompatible Insect Technique)

Successful trial combination (SIT/IIT) trial in China (2019)

66
Q

Tell me some genetic approaches…

A

Oxitec Self-Limiting Gene technology

  • Males require tetracycline in diet to grow to maturity
  • Release males into the wild and mate with females
  • Offspring cannot mature due to lack of tetracycline

Gene Drives:

  • Use CRISPR-Cas9 cassette to force incorporation of defective doublesex gene in offspring (makes females infertile)
67
Q

What are the different stages of the malarial life cycle that drugs can target?

A
  • Target the sporozoites
  • Target the liver forms
  • Target blood forms
  • Target Gametocytes
68
Q

What are some challenges with the malarial vaccine?

Whats an alternative to possibly overcome these challenges?

A

Challenge: malaria isn’t one disease — it’s four!

Complexity of life cycle? Which stage to target?

Generate enough antibody to eliminate every single sporozoite before they reach the liver, as a single parasite can expand by 10,000 to 40,000 times!

Immune evasion by hiding in host cells

Most vaccines designed to target the infectious stage — the single-celled sporozoites that are injected into the body by a feeding mosquito

Alternative is to develop a vaccine against the transmission stage of life cycle (use in combination)

69
Q

What are the current and future malarial vaccines?

A

Current and future vaccines

  • Bill & Melinda Gates Foundation/GSK: RTS,S/AS01 (Mosquirix®) for P. falciparum
  • Recombinant protein antigen (circumsporozoite protein antigens, fusion with hepatitis B surface antigen)
  • Provides protection for both clinical and severe malaria in African children (with about a 55 % effectiveness over 12 months)
  • Might be more effective with targeted roll out (i.e. rainy season)
  • Requires 4 doses (3, per month and 1 at 18 months)
  • Concerns over cost (lack of support for other programmes)
  • Next generation: R21/MM has an efficacy of 75% (phase II)
70
Q

When are drugs taken for prophylaxis/ acute attacks?

A

Generally taken daily or weekly (at lower dose) and restricted to short term visitors

71
Q

Why are drugs for prophylaxis/ acute attacks generally not taken in endemic regions, explain a bit more about this

A

Often not used in endemic regions due to cost / side effects

Either causal (liver stage; ‘liver schizonticides’; required week upon return)

or

suppressive (blood stage, ‘blood schizonticides’; required month upon return)

72
Q

What are the major drug targets for those targeting prophylaxis/ acute attacks?

A

Folate metabolism

Mitochondrial electron transport

Haem detoxification, reactive oxygen species

73
Q

What being used in combination has also proven useful for prophylaxis/ acute attacks?

A

Some antibiotics, such as doxycycline and clindamycin, have also proved useful in combination

74
Q

Give an example of drugs that targets liver forms (hypnozoites) - drugs that effect a radical cure

Tell me about it amd when they are active against

A

8-aminoquinoline: ‘tissue schizonticide’ active against the exoerythrocytic stage of the parasite AND active against liver hypnozoites (the dormant stage in P. ovale and P. vivax)

Helps to prevent relapse (that occurs sometimes weeks, or even years after infection) in combination with chloroquine

Active against gametocytes (of all four species) and helps reduce the spread of infection

MOA not known, not much resistance

Tafenoquine also approved in 2018 (70 years after primaquine!)

75
Q

During the exo/erythrocytoc stage, what must the parasite do?

A

rapidly divide and synthesis folate

76
Q

What are dihydrofolate derivates essential cofactors for?

A

single carbon transfer reactios in the synthesis of nucleic acids and methionine

77
Q

Rapidly dividing parasites, such as those that reproduce asexually rely heavily on what?

Compare this to what humans rely on

A

Rapidly dividing parasites rely on the availability of such folates for growth

Whereas humans rely on the dietary intake of dihydrofolate, Plasmodium are capable of its de novo biosynthesis from dihydropteroate, p-amino benzoic acid (pABA) and glutamate

78
Q

Tell me the steps to folate metabolism and the drugs that can act on each stage of this metabolism

A
79
Q

In what way do structrual analogues act?

A

By competitive inhibition

80
Q

Tell me the following about structural analogues…

Wha are they used in combination with?

Examples?

A

Most used in combination with other drugs (resistance) e.g. pyrimethamine and sulphadoxine (Fansidar ®)

Proguanil – prodrug -> cycloguanil. Used in combination with atovaquone (see later)

Dapsone – too many side effects!

81
Q

Atovaquone (a drug used to treat PCP) targets what?

A

electron transport

82
Q

Tell me the following about Atovaquone…

  • What type of inhibitor is it
  • What does it inhibit
  • Whats its selective toxicity based on
  • What is it functional against
A

Competitive inhibitor of ubiquinone (UQ)

inhibits mitochondrial electron transport chain at the cytochrome bc1 complex

collapsing membrane potential

loss of mitochondrial function (may also inhibit purine biosynthesis)

Selective toxicity based on much higher affinity for parasite complex (IC50 µM vs nM)

Functional against many metabolising stages of the parasite in host and mosquito (not hypnozoites)

83
Q

What is the combination therapy of Malarone a combination of?

A

(Proguanil/Atovaquone)

84
Q

Tell me a bit about the combination therapy of Malarone

  • when is it given in combination with?
  • Whats it used for
A
  • Atovaquone is almost always given in combination with proguanil (5:2) as the two act synergistically (proguanil does not act as a DHFR inhibitor – mechanism?!)
  • Very useful prophylactic (UK (70% of all prescriptions): can be started 1-2 days before traveling to an area where malaria transmission occurs 7 days after traveling
  • Also used for treating chloroquine-resistant malaria (complex)
  • GSK lost the patent in 2014 on account of obviousness (much cheaper now!)
85
Q

With drugs that inhibit haem detoxification (quinolines) what is their selective toxicity due to?

A

Selective toxicity is due to targeting haem biocrystallisation AND that they selectively accumulate in the Plasmodiumdigestive vacuole where this occurs

86
Q

During the erythocytic stage, what is a major nutrient source?

A

Haemoglobin

87
Q

During a parasites intraerythrocytic asexual reproduction cycle what does it digest up to 80% of?

A

The RBCs haemoglobin to provide a source of amino acids (catabolism)

88
Q

Tell me the following about parasitic digestion during the erythrocytic stage…

Where does it occur

What does it release

What is toxic and why

What can it bind to

A

During its intraerythrocytic asexual reproduction cycle parasite digests up to 80% of the RBCs haemoglobin to provide a source of amino acids (catabolism)

Digestion occurs in the digestive vacuole (low pH + proteases)

Releases monomeric haem (a ferriprotoporphyrin) / peptides + amino acids

Haem is toxic since it is a pro-oxidant and catalyses the production of reactive oxygen species (no haem oxygenase like us)

It can also bind to and disrupt cell membranes, damaging cell structures and causing the lysis of the host erythrocyte

Plasmodium have developed a clever strategy to remove the haem

89
Q

Haem detoxification by biocrystallisation into what?

A

Hemozoin

90
Q

Structure of hemozoin crystals

A
91
Q

What happens if you prevent biocrystallisation?

A

Prevent biocrystallisation and lead to build up of toxic haem (e.g., CQ)

92
Q

Why is there an accumulate in the digestive vacuole (e.g., CQ)?

What are the concentrations?

What is it due to?

Tell me about the deprotonated form of CQ

A

Concentrate from nanomolar (10-9 M) levels outside the parasite to levels one million times higher (millimolar levels, 10-3 M)

Due to protonation of the exocyclic and ring nitrogen’s of CQ (pKa is around 8.5) and the difference in pH inside the digestive vacuole (pH 4.5) compared to outside (pH 7.5)

Deprotonated form of CQ is membrane permeable, whereas the charged (protonated) form is not

NOTE: CQ (chloroquine) is used to prevent and treat malaria and amebiasis (an infection of the intestine with a parasite called E. histolytica)

93
Q

Whats is Artemisinin isolated from?

What is it used in?

Tell me about its modern synthesis and what it is now used to treat

A

Artemisia annua (sweet wormwood)

Used in chinese traditional medicine: “emergency prescriptions kept up one’s sleeve” Ge Hong

Rediscovered in 1970s by Tu Youyou who was awarded 2015 Nobel Prize for Medicine

Modern synthesis: recombinant yeast to make precursor molecule via an engineered mevalonate pathway so costs reduced to 0.25 US cents/dose !

Primary treatment for severe malaria (intravenous) P. falciparum

94
Q

Tell me about some semi-synthetic derivatives of Artemisinin and what their properties

A
95
Q

What is the proposed mechanism of action of Artemisinin?

Tell me the reaction equation?

A

Cleavage of endoperoxide bridge by free heme/Fe2+ produces reactive oxygen species:

Directly alkylates proteins = protein miss function

Also, a direct inhibitor of phosphatidylinositol-3-kinase (involved in stress response)

96
Q

What is Artemisinin-based Combination Therapies (ACT) the primary treatment for?

A

symptomatic uncomplicated (oral) malaria (P. falciparum)

97
Q

What is Artemisinin-based Combination Therapies (ACT) not often used for?

Tell me about its possible combination therapy?

A

Not often used for prophylaxis due to short t1/2 (half-life) of drug

Combination therapy – multiple targets to reduce resistance development and make use of longer half-life drugs e.g.

98
Q

What does Artesunate – mefloquine = ?

A

Oxidative stress (protein and hemozoin crystallisation)

99
Q

The following image shows new drug combinations/formulations that have been approved for use

A
100
Q

When did the resistance to Artemisinin first emerge?

A

Resistance first emerged in SE Asia ~ 15 years ago. Now in Africa, India and South America

101
Q

What does PfKelch13 C580Y mutation (and ~20 others) leads to ?

A

reduction in binding to PfPI3K

102
Q

What do increased levels of PfPI3K (overcome inhibition) and increased PI3P help cope with?

A

Unfolded protein response

103
Q

What else can lead to resistance of other antimalarials within the cell host and microbe?

A
104
Q

Whats coming in the antimalarial pipeline?

A

Artefenomel (trioxane) and piperaquine/ferroquine combination

Same mechanisms as previously but more potent and specific (can target resistant strains) – single dose cure?

Monotherapies and combination completed phase II in phase III

What do we target next?

105
Q

Target the apicoplast with antibiotics?

A

Endosymbiotic organelle of algal origin

Fatty acid synthesis, isoprenoid synthesis and possibly others

Has own ribosomes – sensitive to doxycycline

Doxycycline – only prophylactic… other targets in the apicoplast?

106
Q

summary

A
107
Q

Further reading

A

Further reading

  • Reviews:
  • Philips et al. (2017) “Malaria” Nature Reviews Disease Primers volume 3, 17050
  • Duffy & Gorres (2020) “Malaria vaccines since 2000: progress, priorities, products” npj Vaccines5:48
  • Tse E.G et al. (2019) “The past, present and future of anti‑malarial medicines” Malar J (2019) 18:93
  • Belete T.M. (2020) “Recent Progress in the Development of New Antimalarial Drugs with Novel Targets” Drug Design, Development and Therapy 14 3875–3889
  • Ross & Fidock (2019) “Elucidating Mechanisms of Drug-Resistant Plasmodium falciparum” Cell Host & Microbe 26, July 10, 2019
  • Some web resources:
  • www.who.int/malaria/en/ (WHO)
  • www.malariavaccine.org (RTS,S vaccine)
  • www.gatesfoundation.org/What-We-Do/Global-Health/Malaria (Vaccine)
108
Q

How do the chemical agents of larvicides lead to larval static states of the parasites?

Whats a down side to this chemical?

A

Reduction in the juvenile hormone analogue is linked to the maturation step so if you can artificially raise levels by using analogue which mimics the hormone then the mosquito larvae don’t mature properly- a larval static

Better used in places where we know stagnant water is going to form

Shown to be potentially toxic to aquatic life

109
Q

What do the biological agents of larvicides effect of parasites?

A

It effects the growth of the mosquito and so these are added to certain water sources in order to prevent growth

110
Q

Where are the biodegradable oils of larvicides added and why?

A

These are applied to the surface of the water to stop the larvae from brething

111
Q

What are these two structures shown?

Tell me about the product formed

A

The left shows DDT and on the right shows DDE

DDE is extremely stable and hence why bioaccumulates form in animals

112
Q

There is a large scale resistance to DDT but when is it sometimes used?

A

Sometimes its still used in vector control even though there is a large scale resistance to it (e.g., the Stockholm convention)

113
Q

With the insecticides Pyrethrin and pyrethrois which on is natural and synthetic?

A

Pyrethrin is the natural version

Pyrethroid is the synthetic version

114
Q

What is the benefit of using pyrethrin/ pyrethroids over DDT as an insecticide?

A

They degrade quicker so the toxicity is less than DDT

115
Q

What is one of the downsides of using the pyrethrin/ pyrethroids lots?

A

Large scale use of them has led to resistance

116
Q

What is one of the most targeted enzymes in any toxic organism?

A

Acetylcholine esterase inhibitors

117
Q

Tell me about organophosphates, what do they mimic and hence how does this help with their use?

Are they toxic?

A

Mimics part of the Ach compounds and goes into the active site, reacts with serine in the active site which reversibly inhibits the enzyme

These are toxic as are not selective

Even though toxic they are still used around the world (have killed a lot of people)

118
Q

What do carbamates bind to?

A

Carbamates bind to the same enzyme as the organophosphates i.e., serine

119
Q

Tell me about Acetylcholine agonists at nAChRs

A

Selective agonist for nicotinic Ach receptors

Build-up of ACH In synapse that binds to muscle receptors and leads to muscle spasms and paralysis

Breakthrough as an insecticide (but an article published shows resistance against it still)

120
Q

How does the sterile insect technique on mosquitos affect the male population?

A

Using this on the male population makes them less fit to mate as they have been exposed to ionising radiation so it makes them less competitive by comparioson to those who haven’t been exposed

121
Q

How long has the sterile insect technique been around and what is the main aim?

A

Its been around since the 50s and 60s and the aim is to reduce the population and eventually eradicate the parasite

This was the case on one of the islands, however this island was small and had a small population so was easier to do then with a large island

122
Q

What genetic approach is used more, Oxitec self-limiting gene technology or gene drives?

A

Gene drives are used more

123
Q

What happens with the genetic approach, gene drive?

A

Use non-homologous recombination of a defective gene which is important for sex determination of mosquito

124
Q

Tell me what the drug targets are when designing drugs to targets parasites

Which ones are the primary vaccine target and which ones are the hypnozoites?

A

The drug targets:

  • Sporozoites
  • Liver forms
  • Blood forms
  • Gametocytes

The primary vaccine target is: Sporozoites

The Hypnozoites is: the liver forms

The most obviousk place to targte is the blood forms

125
Q

What is currently the only vaccine in use as an anti-malarial?

A

The Bill & Melinda Gates Foundation/GSK: RTS,S/AS01 (Mosquirix®) for P. falciparum-

This species is targeted because it is the most deadly

126
Q

Tell me the use of the hepatitis B surface antigen?

A

Its required by the parasite to get into liver cells at the early stages of the life cycle

Its used with parts of the Hep B virus via surface antigen

Generates virus like particle with antigen on the surface

127
Q

What is the main problem with the current vaccines for malaria?

A

Its expensive as lots of doses are required

Also best to give it at certain times of year e.g, in the rainy season when more mosquitos are present

128
Q

Why can’t drugs for prophylaxis/ acute attacks be taken over a long period of time?

A

They have bad side effects associated with them such as headaches, nausea, etc.,

129
Q

What are the two ways in which the drugs for prophylaxis/ acute attack can be used?

A

They can be used for either causal reasons where they are targeting the earlier stages

Or they can be used for suppressive reasons where they are trying to suppress the symptoms already present

130
Q

Tell me some of the major drug targets for prophylaxis/ acute attacks and why each of these are considered decent targets?

A

Folate metabolism- plasmodium needs lots of folate in order to synthesise dna and replicate rapidly

Mitochondrial electron transport- plasmodia only has one mitochondrion so makes it a good target

Haem detoxification, reactive oxygen species- cause them to kill themselves

131
Q

What is 8-aminoquinoline active against when being used to target liver forms?

A

Active against most of the metabolising stages of the parasite

132
Q

What is the combination therapy of Malarone used for?

A

Infection and it has the least side effects associated with it

133
Q

What do the quinolines all target and what is this?

How do parasites like moasquitos use this to avoid destruction?

Give an example of a quinoline antimalarial that inhibits this process of heme biocrystallisation

A

These all target the metabolism of haemoglobin by targeting haem biocrystallisation

Biocrystallisation is the formation of crystals from organic macromoleucles by living organisms

Blood feeding organisms such as mosquitos digest haemoglobin and release high quantities of free toxic haem

To avoid destruction by this molecule, the parasite biocrystallises heme to form hemozoin

Heme biocrystallisation is inhibited by quinoline antimalarials such as chloroquine

134
Q

What is the structure of the hemozoin crystals?

A

Heme is a prosthetic group consisting of an iron atom (Fe2+) contained in the centre of a heterocyclic porphyrin ring

beta-hematin crystals are made of dimers of hematin molecules that are joined via hydrogen bonds to form larger structures.

In these dimers, an iron-oxygen coordinate bond links the centre iron of one hematin to the oxygen of the carboxylate side chain of the adjacent hematin

Hematin can be a cyclic dimer or a linear polymer

135
Q

It what time frames are Artemisinin-based Combination Therapies (ACT) useful?

A

Only effective in short bursts due to negative side effects

136
Q

Whats the role of the kinases as resitance to artemisinin?

A

The kinases are involved in protecting the cell from oxidative damage

137
Q
A