Photo: Adam Ringham
DNA Diligence – Connect the Dots – Reverse Engineer
The antimalarial drugs chloroquine, hydroxychloroquine, and quinacrine are derivatives of quinine, a natural alkaloid isolated from the South American cinchona bark tree.
So right out of the gate, we see that Hydroxychloroquine is literally derived from cinchona bark!
Just recently, within 20 minutes of doctors sharing a live video of their experience treating COVID19 with hydroxychloroquine- Facebook, YouTube, and Google pulled the video.
Watch the censored video here:
So what makes hydroxychloroquine work?
Recent studies demonstrated that these compounds function as inhibitors of TLR7, TLR8, and TLR9.
We will discuss Toll-Like receptor 9 (TLR9) below in detail.
Hydroxychloroquine was approved for medical use in the United States in 1955. It is on the World Health Organization's List of Essential Medicines, the safest and most effective medicines needed in a health system.
It is a medication used to prevent and treat malaria in areas where malaria remains sensitive to chloroquine. Other uses include treatment of rheumatoid arthritis, lupus, and porphyria cutanea tarda.
Let’s dissect how Hydroxychloroquine works in order to unearth other possible natural remedies, as opposed to the use of pharmaceuticals, which often times are riddled with side effects.
Common side effects of Hydroxychloroquine may include vomiting, headache, changes in vision, and muscle weakness. Severe side effects may include allergic reactions, vision problems, and heart problems.
Serious reported neuropsychiatric adverse effects of hydroxychloroquine use include agitation, mania, difficulty sleeping, hallucinations, psychosis, catatonia, paranoia, depression, and suicidal thoughts.
Pharmacology & Mechanism of action of Hydroxychloroquine
Hydroxychloroquine inhibits stimulation of the toll-like receptor (TLR) 9 family receptors.
TLRs are cellular receptors for microbial products that induce inflammatory responses through activation of the innate immune system.
Hydroxychloroquine has similar pharmacokinetics to chloroquine, with rapid gastrointestinal absorption, large distribution volume, and elimination by the kidneys. Cytochrome P450 enzymes (CYP2D6, 2C8, 3A4 and 3A5) metabolize hydroxychloroquine to N-desethylhydroxychloroquine.
Both agents also inhibit CYP2D6 activity and may interact with other medications that depend on this enzyme.
Hydroxychloroquine increases lysosomal pH in antigen-presenting cells. In inflammatory conditions, it blocks toll-like receptors on plasmacytoid dendritic cells (PDCs).
Toll-like receptor 9 (TLR 9), which recognizes DNA-containing immune complexes, leads to the production of interferon and causes the dendritic cells to mature and present antigen to T cells.
Hydroxychloroquine, by decreasing TLR signaling, reduces the activation of dendritic cells and the inflammatory process.
Pathogens commonly utilize endocytic (clathrin) pathways to gain cellular access.
The endosomal pattern recognition receptors TLR7 and TLR9 detect pathogen-encoded nucleic acids to initiate MyD88- dependent proinflammatory responses to infection.
Hydroxychloroquine has been suggested to be a zinc ionophore and may derive an anti-cancer action from increasing intracellular zinc uptake.
CYP2D6 & Toll-like receptor 9 (TLR 9) seem to be the two main genes involved with the Pharmacology & Mechanism of action behind Hydroxychloroquine.
Cytochrome P450 2D6 (CYP2D6) is an enzyme that in humans is encoded by the CYP2D6 gene. CYP2D6 is primarily expressed in the liver. It is also highly expressed in areas of the central nervous system, including the substantia nigra.
CYP2D6, a member of the cytochrome P450 mixed-function oxidase system, is one of the most important enzymes involved in the metabolism of xenobiotics in the body.
In particular, CYP2D6 is responsible for the metabolism and elimination of approximately 25% of clinically used drugs.
Hydroxychloroquine inhibits CYP2D6 so this will be the main focus for natural discovery.
Natural Inhibitors of CYP2D6:
· Cannabidiol (STRONG)
· St. John’s Wort
· Vitamin B3
· Sesame (seeds, oil)
CYP2D6 cytochrome P450 is involved in the metabolism of fatty acids, steroids and retinoids. Mechanistically, uses molecular oxygen inserting one oxygen atom into a substrate, and reducing the second into a water molecule, with two electrons provided by NADPH via cytochrome P450 reductase (NADPH--hemoprotein reductase).
Metabolizes endocannabinoid arachidonoylethanolamide (anandamide) to 20-hydroxyeicosatetraenoic acid ethanolamide (20-HETE-EA) and 8,9-, 11,12-, and 14,15-epoxyeicosatrienoic acid ethanolamides (EpETrE-EAs), potentially modulating endocannabinoid system signaling.
Metabolizes cholesterol toward 25-hydroxycholesterol, a physiological regulator of cellular cholesterol homeostasis. Catalyzes the oxidative transformations of all-trans retinol to all-trans retinal, a precursor for the active form all-trans-retinoic acid (Vitamin A).
Compounds for CYP2D6 Gene:
· St. John's Wort
· Linoleic acid
· Vitamin A
· Vitamin D3
· Black cohosh
· Rhubarb (Rhein)
· Arachidonic acid
· Stinging Nettle (Formic acid)
Toll-like receptor 9 (TLR 9)
TLR9 is an important receptor expressed in immune system cells including dendritic cells, macrophages, natural killer cells, and other antigen-presenting cells.
TLR9 preferentially binds DNA present in bacteria and viruses and triggers signaling cascades that lead to a pro-inflammatory cytokine response.
Cancer, infection, and tissue damage can all modulate TLR9 expression and activation.
TLR9 is also an important factor in autoimmune diseases, and there is active research into synthetic TLR9 agonists and antagonists that help regulate autoimmune inflammation.
The TLR family plays a fundamental role in pathogen recognition and activation of innate immunity.
TLR9 is preferentially expressed in immune cell-rich tissues, such as spleen, lymph node, bone marrow and peripheral blood leukocytes. Studies in mice and humans indicate that this receptor mediates cellular response to unmethylated CpG dinucleotides in bacterial DNA to mount an innate immune response.
TLR9 interacts with MyD88, the primary protein in its signaling pathway.
TLR9 can work through MyD88, an adaptor molecule that increases the expression of NF-κB.
Therefore, NF-κB inhibition is also vital!
Compounds for NFKB1 Gene:
· Fish oil
· Caffeic acid phenethyl ester (Bee propolis)
· Cardamonin (Ginger family)
· Tanshinone IIA (Dan shen)
· Matrine (Sophora)
· Withaferin A (Ashwagandha)
TLR9 signals leads to activation of the cells initiating pro-inflammatory reactions that result in the production of cytokines such as type-I interferon, IL-6, TNF and IL-12.
Toll-Like Receptor 9 (TLR9) is a unique pattern recognition receptor due to its ability to induce either pro- or anti-inflammatory cascades.
TLR9 is a nucleotide-sensing TLR which is activated by unmethylated cytidine-phosphate-guanosine (CpG) dinucleotides.
Acts via MYD88 and TRAF6, leading to NF-kappa-B activation, cytokine secretion and the inflammatory response.
TLR9 interacts with both Heat Shock Proteins 70/90.
TLR9 also interacts with SMPDL3B (Sphingomyelin Phosphodiesterase Acid Like 3B) which acts as a negative regulator of Toll-like receptor signaling and can cleave CDP-choline, and can release phosphate from ATP and ADP.
In other words, CDP-choline helps regulate SMPDL3B which then helps to regulate TLR9.
For optimal receptor signaling, it’s proteolytically processed by first removing the majority of the ectodomain by either asparagine endopeptidase (AEP) or a cathepsin followed by a trimming event that is solely cathepsin mediated.
The gene card for TLR9, under drugs and compounds, we see that Hydroxychloroquine with its “mechanism of action” includes an Autophagy inhibitor.
Compounds for TLR9 Gene:
· Liver Extracts
· Hydroxychloroquine - Mechanism of Action - Autophagy inhibitor
· Tumor necrosis factor alpha (TNF-alpha) inhibitors:
o Glucosamine (NAG)
o Glycyrrhizin (licorice)
o Tanshinone IIA (dan shen)
Autophagy means (“self-eating”).
Natural Compounds from Herbs that can Potentially Execute as Autophagy (Study)
Natural Autophagy Inhibitor from Herbs:
· Betulinic acid (Prunella vulgaris)
· Epigallocatechin gallate (EGCG)
Natural agents as modulators of autophagic signals:
· Tanshinone IIA (dan shen)
· Ursolic acid (prunella vulgaris)
· Glabridin (licorice)
· -Gingerol (ginger)
· Piperlonguminine (Piper longum)
· Oridonin (Rabdosia rubescens)
Tanshinone IIA modulates the initiation of phagophore formation. Ginsensoside and ursolic acid affect the formation of autophagosomes. Ginsenoside RO (Panax ginseng) inhibits autophagosome–lysosome fusion.
Modulation of Toll-Like Receptor Signaling in Innate Immunity by Natural Products (Study)
Linking Natural Products to Innate Immunity
TLRs are highly conserved PRRs that activate the innate immune system and participate in initiating the inflammatory response. When TLR activity is dysregulated, there is an increased risk of developing chronic inflammatory and immune diseases.
Studies have shown that diets rich in fruits and vegetables are associated with lowered risk of cardiovascular disease and other inflammatory disorders.
Many of the phytochemicals found in fruits and vegetables have beneficial anti-inflammatory actions in cells. Therefore, it is possible that these phytochemicals exert their mechanism of action by targeting TLRs and their signaling molecules downstream.
Recently, Yi et al. reported that ω-3 polyunsaturated fatty acids (PUFAs), abundant in nuts, oils, and fish, suppress the excessive inflammation in patients with severe trauma via a signaling pathway mediated by TLRs and NF-κB.
The group found that levels of COX-2, IL-2, and TNF-α substantially decreased in these patients. Additionally, Landmann et al. discovered that pretreating mice with chicoric acid found in the plant Echinacea purpurea attenuated the harmful effects of alcohol on the liver through suppression of mRNA expression of TNF-α and inducible nitric oxide synthase (iNOS). Currently, various natural compounds and their derivatives were found to act as agonists or antagonists for TLR family members and their downstream signaling molecules.
Agonist activity is shown in green while antagonist or inhibitory activity is shown in red.
In the image, we see that DHA, Rhein (Rhubarb), Emodin (Sea buckthorn etc.), Kukoamine B (cortex Lycii/Wolfberry Root), and Lyngbya majuscula, commonly termed “mermaids hair” or “fireweed, target Toll-Like Receptor 9.
Artemisinin possesses potent antiproliferative and anti-inflammatory properties associated with toll-like receptor signaling.
Mustard seed from the mustard plant is a popular food seasoning worldwide, especially in Japan, India, and China. Mustard seed possesses several biological effects, including anti-inflammatory, antioxidant, and antitumor effects. Mustard seed inhibited the infiltration of various types of leukocytes, including DCs, macrophages, and T cells.
Curcumin might inhibit endosomal TLR-induced inflammation by targeting NF-κB signaling.
Resveratrol inhibited imiquimod-induced expression of IL-17A, IL-19, and IL-23p19. Resveratrol has been shown to inhibit LPS- and TNF-α-induced NF-κB activation.
Andrographis possesses anti-inflammatory activity and promoted MyD88 degradation and blocked the recruitment of TRAF6 to form signalsomes.
TLR9 Interactive DNA String Network
Held within this genes network there is a gene called SYK (Spleen Tyrosine Kinase).
SYK, along with Zap-70, is a member of the Syk family of tyrosine kinases.
Syk is an essential regulator of lymphatic system development.
In the current COVID-19 pandemic, drug repurposing is being widely investigated as a way to find treatments rapidly. Now, a new study addresses the possible use of an FDA-approved monoclonal antibody, called fostamatinib, a spleen tyrosine kinase inhibitor (SYK), to reduce the levels of mucin-1, a molecule associated with acute lung injury and acute respiratory distress syndrome (ARDS). People who have severe COVID-19 disease may develop ARDS.
Several transforming viruses contain "Immunoreceptor Tyrosine Activation Motifs" (ITAMs) which lead to activation of Syk including Epstein Barr virus, bovine leukemia virus, and mouse mammary tumor virus.
MERS-CoV replication significantly upregulated C-type lectin receptor (CLR) macrophage-inducible Ca2+-dependent lectin receptor (Mincle). The role of Mincle for MERS-CoV-triggered cytokine/chemokine induction was established based on the results of antibody blockage, siRNA depletion of Mincle and its adaptor spleen tyrosine kinase (Syk), and Syk pharmacological inhibition.
Spleen tyrosine kinase (SYK) inhibitor, for the treatment of COVID-19 pneumonia.
SYK is a key mediator of immunoreceptor signaling in a host of inflammatory cells. Studies of severe acute respiratory syndrome (SARS) and other acute viral respiratory infections suggest that the pathogenesis relies on a series of SYK-dependent events involving activation of C-type lectin receptors (CLR) and immunoglobulin Fcg receptors (FcgR) in multiple cell types. Such SYK-mediated processes result in excessive cytokine and chemokine release, neutrophil activation associated with extensive NETosis (a highly inflammatory and thrombogenic type of cell death), and endothelial cell stimulation leading to vascular endothelium leakage and edema in the lungs. Together, these events can contribute to acute respiratory distress syndrome (ARDS), micro-thrombosis and associated systemic complications.
The hallmark of severe COVID-19 is hypoxemia and a radiological pattern of acute lung injury (ALI) that shares features with ARDS. Inhibiting SYK may specifically inhibit the infiltration and activation of monocytes and neutrophils in the lungs that are prominent in COVID-19.
Natural SKY Inhibitors:
· Piceatannol, a natural stilbene (Resveratrol)
· Tanshinone I (Dan Shen)
· Saussurea lappa
· Angelica decursiva
· Chinese skullcap
· White mulberry
· Burdock root
Remember that Hydroxychloroquine works by targeting Toll-Like receptor 9 which is intimately involved with “unmethylated CpG dinucleotides”.
TLR9 detects unmethylated CpG motifs present in viral or bacterial DNA.
Syk is indispensable for CpG-induced activation and differentiation of human B cells (Study)
“B cells are efficiently activated by CpG oligodeoxynucleotides (ODNs) to produce pro-inflammatory cytokines and antibody (Ab). Here, we describe a so far unidentified, spleen tyrosine kinase (Syk)-dependent pathway, which is indispensable for CpG-induced human B cell activation. We show that triggering of B cells by CpG results in Syk and src kinase phosphorylation, proliferation, as well as cytokine and Ab production independent of the BCR. Notably, all these functions are abrogated when Syk is inhibited. We demonstrate that CpG-induced Syk activation originates from the cell surface in a TLR9-dependent manner. While inhibition of Syk does not influence the uptake of CpG ODNs, activation of the kinase is a prerequisite for the delivery of CpG into TLR9-containing endolysosomes and for the CpG-induced up-regulation of TLR9 expression. Our results reveal an alternative, Syk-dependent pathway of CpG-induced B cell stimulation, which is initiated at the plasma membrane and seems to be an upstream requirement for endosomal TLR9-driven B cell proliferation and differentiation.”
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