NEW APPROACHES FOR THE MAJOR INFECTIVE DISEASES

Knox Van Dyke (Prof. West Virginia University Knox Van Dyke (Prof. West Virginia University U.S.A.)U.S.A.)

Prof. Erhabor Osaro (Usmanu Danfidiyo Prof. Erhabor Osaro (Usmanu Danfidiyo University, Sokoto, Nigeria)University, Sokoto, Nigeria)

HIV ,TB and MalariaThese are the major infectious diseases (killer 3). These are the major infectious diseases (killer 3).

Major cause of death & non-productivity in SSA.Major cause of death & non-productivity in SSA.

Available treatments have been chemotherapy.Available treatments have been chemotherapy.

They are expensive and prone to drug resistance.They are expensive and prone to drug resistance.

We are always sourcing new We are always sourcing new

drugs/combinations.drugs/combinations.

The Triad of InfectionAll infectious diseases are affected by 3

factors: Infectious agent (bug). The host factors (most important factor). The drug/s being used for treatment. Interaction between the drug & the bug is

presently exploited. There may be need to start targeting the

host immune system.

HIV and TB Co-infectionHIV and TB Co-infectionHIV-TB is a globally common co-infection. HIV-TB is a globally common co-infection.

Up to 80% of TB patients are also HIV-positive.Up to 80% of TB patients are also HIV-positive.

Without treatment, HIV & TB can work as a team to Without treatment, HIV & TB can work as a team to

shorten life of the person infected. shorten life of the person infected.

Untreated latent TB & HIV infection likely result in active Untreated latent TB & HIV infection likely result in active

TB (than without HIV). TB (than without HIV).

HIV facilitates progression from latent to active TB.HIV facilitates progression from latent to active TB.

HIV infection & active TB (AIDS-defining). HIV infection & active TB (AIDS-defining).

Changing The Host Factors: Could They Changing The Host Factors: Could They Make a Major Difference?Make a Major Difference?

HIV-1 is the number 1 infectious dx in today’s world. HIV-1 is the number 1 infectious dx in today’s world.

Schreck & Baeurele in Germany has shown that oxidative Schreck & Baeurele in Germany has shown that oxidative

mechanisms (peroxynitrite & hypochlorite can mechanisms (peroxynitrite & hypochlorite can

upregulate HIV infection). upregulate HIV infection).

Van Dyke indicates has shown that a series of Van Dyke indicates has shown that a series of

inexpensive antioxidants and anti-inflammatory steroids inexpensive antioxidants and anti-inflammatory steroids

(inhibit HIV & cure FIV infection. (inhibit HIV & cure FIV infection.

Antioxidants & Steroids Affect both Virus & HostAntioxidants & Steroids Affect both Virus & Host

Combination of substances was used in FIV infected cats Combination of substances was used in FIV infected cats

(6) over a period of multiple months (4-5) the cats were (6) over a period of multiple months (4-5) the cats were

cured from the retroviral disease. Untreated cat died.cured from the retroviral disease. Untreated cat died.

We found a patient with HIV for 10 years (long term We found a patient with HIV for 10 years (long term

survivor) who had AIDS (weak with signs of dementia). survivor) who had AIDS (weak with signs of dementia).

His physician allowed us to try our His physician allowed us to try our steroid/antioxidantsteroid/antioxidant

combination and his viral load started to drop.combination and his viral load started to drop.

Continuous HIV Treatment

After several months treatment, his dementia subsided, he got better, his appetite returned & he started to gain weight.

Finally after treatment for one year, he displayed undetectable HIV in his blood.

He went to work and held 3 jobs.

Mechanisms of Antioxidant/Steroid

Combination of antioxidants Combination of antioxidants

(Vit C & E) at (Vit C & E) at higher doses higher doses

oppose oxidant- stimulated oppose oxidant- stimulated

replication of HIV.replication of HIV.

Steroids inhibit multiple nf-Steroids inhibit multiple nf-

Kappa B sites on retroviruses.Kappa B sites on retroviruses.

This prevents replication of the This prevents replication of the

virus.virus.

Drug Companies Not InterestedDrug Companies Not InterestedDrug companies are not interested Drug companies are not interested

Investment costs & low potential returns from profits.Investment costs & low potential returns from profits.

Why? These drugs and supplements are inexpensive & Why? These drugs and supplements are inexpensive &

the rigorous research work in humans would be too the rigorous research work in humans would be too

expensive to turn a profit.expensive to turn a profit.

This could be a perfect solution for SSA countries. This could be a perfect solution for SSA countries.

SSA could save precious funds to treat more people.SSA could save precious funds to treat more people.

Malaria: The Oldest Infectious DiseaseFalciparum malaria is a major Falciparum malaria is a major

infectious diseases. infectious diseases.

Malaria kills millions of people /year.Malaria kills millions of people /year.

Hundreds of thousands of innocent Hundreds of thousands of innocent

children & pregnant women have children & pregnant women have

died over the years from malaria. died over the years from malaria.

Malaria was the major cause of Malaria was the major cause of

soldier’s death in the Vietnam war. soldier’s death in the Vietnam war.

Malaria Control ChallengesMalaria Control ChallengesEstimated 500m cases of malaria per year. Estimated 500m cases of malaria per year.

Unless new strategies are deployed, the health and Unless new strategies are deployed, the health and

economic burden will worsen. economic burden will worsen.

Main obstacle to malaria control is resistance. Main obstacle to malaria control is resistance.

Combinations therapy reduces risk of resistance.Combinations therapy reduces risk of resistance.

Combination of an artemisinin derivative & mefloquine Combination of an artemisinin derivative & mefloquine

(unrelated mode of action).(unrelated mode of action).

This strategy seems the way forward (for now). This strategy seems the way forward (for now).

Which way with ChemotherapeuticsWhich way with Chemotherapeutics ChloroquineChloroquine

QuinineQuinine

Sulfadoxine/perimethamineSulfadoxine/perimethamine

MefloquineMefloquine

DoxycyclineDoxycycline

AtovaquoneAtovaquone

ProguanilProguanil

Artemisinin-Combination Therapy (ACT) Artemisinin-Combination Therapy (ACT)

Amodiaquine, Lumefantrine, mefloquine Amodiaquine, Lumefantrine, mefloquine

or Sulfadoxine/perimethamineor Sulfadoxine/perimethamine

Dihydroartemisinin and PiperaquineDihydroartemisinin and Piperaquine

Development of First High Throughput Antimalarial Development of First High Throughput Antimalarial Screening SystemScreening System

In 1966 Knox was assigned the task of developing a new in vitro, antimalarial screening system.

He incubated radioactive (3H) adenosine with parasitized cells & found certain drugs inhibited the incorporation of the radioactive nucleotide into parasite nucleic acids.

Further research showed that the parasite actually converted adenosine to the free oxidized base hypoxanthine.

Knox’sKnox’s high throughput screening system for antimalarial high throughput screening system for antimalarial

drugs effectiveness using a quick DNA/RNA labeling system.drugs effectiveness using a quick DNA/RNA labeling system.

Ye & Van Dyke used this DNA/RNA screening system to Ye & Van Dyke used this DNA/RNA screening system to

demonstrate that chloroquine resistance can be entirely demonstrate that chloroquine resistance can be entirely

overcome in vitro & in vivo by adding overcome in vitro & in vivo by adding

bisbenzylisoquinolinesbisbenzylisoquinolines to chloroquine. to chloroquine.

Thus for the first time choroquine sensitivity can be Thus for the first time choroquine sensitivity can be

restored!restored!

Trager /Jensen falciparum culture 3H Hypoxanthine was the key label

Can We Assist Host Mechanisms with Drugs to Fight Malaria Can We Assist Host Mechanisms with Drugs to Fight Malaria more effectively?more effectively?

Macrophages are important in the host response to Macrophages are important in the host response to

malaria. malaria.

Its oxidative activity likely stems from peroxynitrite.Its oxidative activity likely stems from peroxynitrite.

Does the parasite affect the macrophages killing mechanism Does the parasite affect the macrophages killing mechanism

making it less effectivemaking it less effective? Yes!? Yes!

The macrophages has two pathways to produce NO.The macrophages has two pathways to produce NO.

It oxidizes L-arginine to L- citrulline and NOIt oxidizes L-arginine to L- citrulline and NO

It recycles L-citrulline to L-arginine. It recycles L-citrulline to L-arginine.

Low Plasma L-Arginine Levels in Cerebral Malaria Low Plasma L-Arginine Levels in Cerebral Malaria

Lopansri et al demonstrated that Tanzanian children with

severe cerebral malaria had low plasma L-arginine levels.

Likely due to presence of parasitic and red cell arginase

which depletes L-arginine.

This decreased NO production greatly.

Decreased production of NO in microglia(brain) or blood

macrophages prevents parasite depletion.

Low plasma L-arginine level is significantly associated with

mortality.

Alternative Pathways for Nitric oxide ProductionAlternative Pathways for Nitric oxide Production

Enzymatic NO synthase production of NO Enzymatic NO synthase production of NO

production can be decreased via depleted substrate. production can be decreased via depleted substrate.

Direct production of NO via enzymes/ chemistry. Direct production of NO via enzymes/ chemistry.

Both nitrate and nitrite can generate significant Both nitrate and nitrite can generate significant

amounts of NO for peroxynitrite killing purposes. amounts of NO for peroxynitrite killing purposes.

Nitrates can be obtained from spinach, beets & Nitrates can be obtained from spinach, beets &

seaweed by ingestion of adequate amounts. seaweed by ingestion of adequate amounts.

New sources of Nitric OxideNitrate is concentrated in the salivary glands.

It is converted in the mouth to nitrite

(reduction).

It is then reduced to NO in the acid stomach.

Compounds are recycled over & over in the

body.

Can we increase NO & assist antimalarial drug ?Can we increase NO & assist antimalarial drug ?

Several scientists have attempted to increase NO in malaria patients Several scientists have attempted to increase NO in malaria patients

by giving l-arginine (IV & orally).by giving l-arginine (IV & orally).

Used various doses & found it ineffective. Used various doses & found it ineffective.

After 1 hour NO falls to minimal levels.After 1 hour NO falls to minimal levels.

The half life of l-arginine is The half life of l-arginine is too short too short and so plasma levels using and so plasma levels using

these systems are ineffective. these systems are ineffective.

L-arginine & L-citrulline need to be given orally in a form that L-arginine & L-citrulline need to be given orally in a form that

maintains more consistent levels.maintains more consistent levels.

Other forms of NO production (nitrate & nitrite) are also importantOther forms of NO production (nitrate & nitrite) are also important . .

Adjunctive Therapy Starts EarlyAdjunctive Therapy Starts Early

Giving supplements in advanced cerebral Giving supplements in advanced cerebral

malaria would likely be ineffective.malaria would likely be ineffective.

Starting the NO producing supplements Starting the NO producing supplements

early in the disease early in the disease seems the way seems the way

forward.forward.

““A stitch in time saves nine”A stitch in time saves nine”

TB and Supplement TreatmentTB and Supplement TreatmentTB is the 2nd most important infectious dx TB is the 2nd most important infectious dx

globally.globally.About 1/3 of the population is estimated to be About 1/3 of the population is estimated to be

infected with TB.infected with TB.People with latent infection are asymptomatic & People with latent infection are asymptomatic &

non- infectious.non- infectious.They are potentially at risk of developing active They are potentially at risk of developing active

infections.infections.

Is L-arginine effective in Active TB?

TB bacteria resides in macrophagesTB bacteria resides in macrophages

Causes lesions in the lung. Causes lesions in the lung.

In active dx, the bacteria exerts a major control against In active dx, the bacteria exerts a major control against

the macrophagic killing (chemical suppression). the macrophagic killing (chemical suppression).

Thus bacteria can reproduce in as well as kill Thus bacteria can reproduce in as well as kill

macrophages.macrophages.

TB inhibits NO production via arginase destruction of l-TB inhibits NO production via arginase destruction of l-

arginine. arginine.

NO Availability is Impaired in TB

NO bioavailability affect mycobacterial NO bioavailability affect mycobacterial

clearance.clearance.

Severity is associated with delayed TB Severity is associated with delayed TB

clearance.clearance.

Hypoargininemia has been demonstrated Hypoargininemia has been demonstrated

in TB. in TB.

L-Arginine has proven useful In supplemental TB L-Arginine has proven useful In supplemental TB TreatmentTreatment

Farrazi (2015) showed that L-arginine is useful as Farrazi (2015) showed that L-arginine is useful as

an adjunctive therapy (1 g/day) in active TB. an adjunctive therapy (1 g/day) in active TB.

He concluded that effects were likely due to He concluded that effects were likely due to

increased NO. increased NO.

Constitutional symptoms (Wt gain & reduction in Constitutional symptoms (Wt gain & reduction in

CRP) were improved despite anaemia and cough.CRP) were improved despite anaemia and cough.

ConclusionsAll of the three killer infectious diseases are linked to All of the three killer infectious diseases are linked to

macrophage activity. macrophage activity.

HIV reproduction is stimulated by strong oxidants and HIV reproduction is stimulated by strong oxidants and

inhibited by antioxidants.inhibited by antioxidants.

Anti-inflammatory steroids block the DNA transcription Anti-inflammatory steroids block the DNA transcription

factor nf Kappa B.factor nf Kappa B.

This blocking has a major effect on HIV & FIV replication.This blocking has a major effect on HIV & FIV replication.

Malaria and TB are Inhibited by OxidantsMalaria and TB are Inhibited by Oxidants

We need to be generating large & continuous We need to be generating large & continuous

amounts of peroxynitrite & other antioxidants amounts of peroxynitrite & other antioxidants

The generation must be early in the disease course.The generation must be early in the disease course.

This way the diseases can be more effectively treated This way the diseases can be more effectively treated

with drugs.with drugs.

Accomplished by giving large and continuous amounts Accomplished by giving large and continuous amounts

of substances that generate NO by different pathways. of substances that generate NO by different pathways.

OxidantsOxidantsNO is a very short lived molecule (seconds).NO is a very short lived molecule (seconds).

It must be generated continuously & at high It must be generated continuously & at high

doses using sustained release precursors.doses using sustained release precursors.

All three of the major NO pathways should be All three of the major NO pathways should be

included so the entire body is involved.included so the entire body is involved.

Some antioxidants can be used that actually Some antioxidants can be used that actually

transport nitric oxide (N-acetyl cysteine).transport nitric oxide (N-acetyl cysteine).

Adjunctive Therapy is the way forward against the killer 3 Adjunctive Therapy is the way forward against the killer 3 Infectious Diseases Infectious Diseases

It makes economic sense to use host mechanisms It makes economic sense to use host mechanisms

to their maximal effectiveness.to their maximal effectiveness.

It is most likely to result in the best outcome.It is most likely to result in the best outcome.

The best approach is a synergistic approachThe best approach is a synergistic approach

Way forward is to use selective chemotherapy Way forward is to use selective chemotherapy

with natural NO producing supplements with natural NO producing supplements

AcknowledgementsAcknowledgementsI will like to acknowledge Prof. Erhabor Osaro & his team I will like to acknowledge Prof. Erhabor Osaro & his team

in Usmanu Danfodiyo University, Sokoto, Nigeriain Usmanu Danfodiyo University, Sokoto, Nigeria

Many thanks for your patienceMany thanks for your patience

I will be glad to entertain any questions you may I will be glad to entertain any questions you may have.have.