HONOKIOL IN THE INTEGRATIVE TREATMENT OF CANCER

INTRODUCTION: HISTORY OF HONOKIOL

Magnolia Bark Extract

Extracts from the tree bark of the genus Magnolia have been used for over 2,000 years in traditional Chinese (as Hou Po) and Japanese medicines (as Saiboku-to) to treat a variety of gastrointestinal disorders, neurological diseases, including anxiety, depression, and seizures TCM properties: Bitter, acrid, warm, aromatic Channels: Large intestine, lung, spleen, stomach

The active ingredients in the extracts have been identified as the bi-phenolic isomers magnolol and honokiol

Magnolol and Honokiol Isomers

As honokiol exists naturally with its structural isomer magnolol, which differs from honokiol only by the position of one hydroxyl group, purification has often been limited

Methods developed since 2006, are now able to simply purify the honokiol active ingredient away from the more predominate less active magnolol

MAGNOLIA OFFICINALIS EXTRACT: 98% PURIFIED

HONOKIOL

Botanical Compound Honokiol in the Treatment of Cancer

Honokiol Properties

Properties Anti-inflammatory Antioxidant Selective Pro-oxidant Anti-Microbial Anti-Tumor (Induces Apoptosis & Cell Cycle Arrest) Anti-Angiogenic Anti-Anxiety/Insomnia Neuroprotective Synergistic Anticancer Effect w/ Multiple Chemotherapy Drugs Crosses the Blood–Brain Barrier, Candidate for the Treatment of Central

Nervous System Primary Tumors and Metastases No Appreciable Toxicity

THE MECHANISMS OF ACTION

Honokiol – Main Mechanisms of Action

Blocks signaling in tumors with defective p53 function and activated Ras by blocking activation of phospholipase D

Induces cyclophilin D, potentiating mitochondrial permeability transition pore and causing death in cancer cells with wild-type p53

Blocks NF-kB activation Inhibits mTORcomplex1 Inhibits ROS production Modulates GABA receptors

IN VIVO AND IN VITRO ANTITUMOR ACTIVITY OF HONOKIOL

Extensive Preclinical Oncology Research has Shown that Honokiol:

Inhibits nitric oxide synthesis and tumor necrosis factor (TNF) expression

Inhibits invasive behavior Down-regulates antiapoptotic protein Bcl-xL Inhibits angiogenesis and tumor growth in vivo Induces caspase dependent apoptosis through

down-regulation of the antiapoptotic protein Mcl-1 Overcomes drug resistance in multiple myeloma

Extensive Preclinical Oncology Research has Shown that Honokiol (Continued):

Potentiates the apoptotic effects of TNF and chemotherapeutic drugs

Suppresses induced osteoclastogenesis Suppresses TNF-induced tumor cell invasion

activity Blocks NF-kB activation induced by various agents Suppresses NF-kB activation in a dose-and time-

dependent manner Inhibits constitutive NF-kB activation

Honokiol Anti-Tumor Gene & Protein Expression Modulation

Up regulated p21 & p27 Tyrosine Phosphatase

SHP-1 Cytosolic Cytochrome C Greater Caspase Activity Bax (pro-apoptotic

protein) Bak (pro-apoptotic

protein)

Glucose-Regulated Protein 78 (GRP78)

Calpain Caspase-3, 8, 9 Poly (ADP-ribose)

Polymerase (PARP) p53 Activation (tumor

suppressor gene)

Honokiol Anti-Tumor Gene & Protein Expression Modulation

Down regulated or inhibited Cyclin D1, D2, E, A1 Cyclin-Dependent Kinases

2, 4, 6 Kinases C-Src Janus-Activated Kinase 1, 2 Nuclear Factor-κB (NF-κB)

and Inducible Nitric Oxide Synthases (iNOS)

HIF-1α Cyclooxygenase-2 (COX-2)

Prostaglandin (PG) E2

Peroxisome Proliferator-Activated Receptors –Gamma (PPAR-Gamma)

Mammalian Target of Rapamycin Complex 1 (mTORC1)

Bcl-xL EGFR Signaling Mitogen-Activated Protein

Kinase Akt Mcl-1

Effects of honokiol (H) on extrinsic (receptor-mediated) & intrinsic (mitochondria-mediated) apoptosis pathways in cancer

HH

Huanli X, et al. Drug Discoveries &

Therapeutics. 2011; 5(5):202-210

The extrinsic apoptosis pathway involves ligation of death receptors with their ligands, resulting in a sequential activation of caspase-8 and -3

Intrinsic death stimuli indirectly activates the mitochondrial pathway by inducing release of cytochrome c and formation of the apoptosome, which consists of Apaf-1 and caspase-9. Caspase-9 is activated at the apoptosome and in turn activates procaspase-3

Regulation of Cancer Cell Metabolism

Nature Reviews Cancer 11, 85-95, 2011

mTOR Signaling

Selected Honokiol Cancer Research

Honokiol traverses the blood-brain barrier and induces apoptosis of neuroblastoma cells via an intrinsic bax-mitochondrion-cytochrome c-caspase protease pathway. Lin JW, et al. Neuro Oncol. 2012 Jan 18

Anti-metastatic activity of honokiol in osteosarcoma. Steinmann P, et al. Cancer. 2011 Sep 20. doi: 10.1002/cncr.26434

Honokiol arrests cell cycle, induces apoptosis, and potentiates the cytotoxic effect of gemcitabine in human pancreatic cancer cells. Arora S, et al. PLoS One. 2011;6(6):e21573

Selected Honokiol Cancer Research

Honokiol synergizes chemotherapy drugs in multidrug resistant breast cancer cells via enhanced apoptosis and additional programmed necrotic death. Tian W, et al. Int J Oncol. 2012 Dec 14. doi:10.3892/ijo.2012.1739

Honokiol produces anti-neoplastic effects on melanoma cells in vitro. Mannal PW, et al. J Surg Oncol. 2011 Sep 1;104(3):260-4

Honokiol radiosensitizes colorectal cancer cells: enhanced activity in cells with mismatch repair defects. He Z, et al. Am J Physiol Gastrointest Liver Physiol. 2011 Nov;301(5):G929-37

Selected Honokiol Cancer Research

Apoptosis induced by Magnolia grandiflora extract in chlorambucil-resistant B-chronic lymphocytic leukemia cells. Marin GH, Mansilla E. J Cancer Res Ther. 2010 Oct-Dec;6(4):463-5

Modulation of multidrug resistance p-glycoprotein activity by flavonoids and honokiol in human doxorubicin-resistant sarcoma cells (MES-SA/DX-5): implications for natural sedatives as chemosensitizing agents in cancer therapy. Angelini A, et al. J Biol Regul Homeost Agents. 2010 Apr-Jun;24(2):197-205

Honokiol induces paraptosis and apoptosis and exhibits schedule-dependent synergy in combination with imatinib in human leukemia cells. Wang Y, Yang Z, Zhao X. Toxicol Mech Methods. 2010 Jun;20(5):234-41

Effect of Honokiol on Growth of PC3 Cells (Pre-Published Data)

Honokiol Reviews

Honokiol, a multifunctional tumor cell death inducer. Tian W, et al. Pharmazie. 2012 Oct;67(10):811-6

Honokiol, a multifunctional antiangiogenic and antitumor agent. Fried LE, Arbiser JL. Antioxid Redox Signal. 2009 May;11(5):1139-48

Effect of Honokiol on the Growth of MDA-MB-231 Cells (Pre-Published Data)

Synergistic Effect of Modified Citrus Pectin (MCP) & 98% Honokiol on PC3 Cell line Migration (Pre-Published Data)

Control MCP 1 mg/ml

MCP 2 mg/ml

MCP 4 mg/ml

%98 Honokiol

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%98 Honokiol

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%Migration of PC3 Cells

Effect of Honokiol on the Growth of 786-0 and A-498 Human Renal Cell Carcinoma 786-0Cells (Pre-Published Data)

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iol

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% Proliferation

EFFECT OF HONOKIOL ON REACTIVE OXYGEN SPECIES

The Inhibition by Honokiol of Reactive Oxygen Driven Tumors

Due to its involvement with the NADPH oxidase (NOX) pathway. This inhibition was first demonstrated

in neutrophils in hepatocytesHuman Umbilical Vein Endothelial Cells (HUVECs)

NF-kB regulates the signaling pathway of NOX-mediated oxidative stress

Honokiol Improves the Functions of Normal Human Cells

Results of oxygen consumption and malondialdehyde (MDA) production showed that the honokiol inhibition was 1,000 times that of α-tocopherol (vitamin E)

As honokiol is better than α-tocopherol at inhibiting lipid peroxidation, it was used to protect the myocardium against ischemic injury by suppressing ventricular arrhythmia during ischemia and reperfusion

Studies have shown the protective effect of honokiol on hepatocytes from peroxidative injury, oxygen consumption, and malondialdehyde formation

Honokiol has Both Pro- and Antioxidant Activities

Honokiol has antioxidant activities because the allyl groups on honokiol can react with reactive oxygen species

Also the anti-mitochondrial effects of honokiol generates reactive oxygen by activating the mitochondrial permeability transition pore. The ability of honokiol to activate the mitochondrial pore may be dependent on the p53 status, with tumors with wild-type p53 having greater susceptibility to pore formation

IMMUNE & ANTI-INFLAMMATORY EFFECTS OF HONOKIOL

Targeting the Intrinsic Inflammatory Pathway: Honokiol Exerts Proapoptotic Effects Through STAT3 Inhibition in Transformed Barrett's Cells

Yu C, et al. Am J Physiol Gastrointest Liver Physiol. 2012 Sep 1;303(5):G561-9 Signal transducer and activator of transcription 3 (STAT3)

contributes to the intrinsic inflammatory pathway in Barrett's esophagus.

In human tumors, honokiol has growth-inhibitory and proapoptotic effects associated with suppressed activation of STAT3

Honokiol causes necrosis and apoptosis in transformed Barrett's and esophageal adenocarcinoma cells, but not in nontransformed Barrett's cells, and the proapoptotic effects of honokiol are mediated by its inhibition of STAT3 signaling.

Honokiol, a Natural Plant Product, Inhibits Inflammatory Signals & Alleviates Inflammatory Arthritis

Munroe ME, et al. J Immunol 179: 753–763, 2007 Treatment with honokiol significantly decreased the

clinical scores of collagen-induced arthritis in both normal and transgenic mice

Antibody production, most notably IgG3, was diminished, as were IL-12, IL-6, interferon gamma, and notably IL-17

These findings indicate that honokiol may have benefit against IL-17–mediated inflammatory disorders, including rheumatoid arthritis, psoriasis, and inflammatory bowel disease

NEUROLOGIC EFFECTS OF HONOKIOL

GABA (γ-Aminobutyric acid)

The major inhibitory neurotransmitter in the central nervous system

GABA acts at inhibitory synapses in the brain by binding to specific transmembrane receptors in the plasma membrane of neuronal processes

It is synthesized in the brain from glutamate using the enzyme L-glutamic acid decarboxylase and pyridoxal phosphate (active form of vitamin B6) as a cofactor via a metabolic pathway called the GABA shunt.

Increase in the available amount of GABA typically have relaxing, anti-anxiety, and anti-convulsive effects

GABA is synthesized from glutamate in a reaction catalyzed by glutamic acid decarboxylase (GAD)

"GABA Shunt"

GABA Production Enhancers

Taurine helps breakdown glutamate to GABA Lysine enhances GABA action Manganese is essential for glutamine synthesis Pyridoxine (Vitamin B6) is an essential cofactor for

the GABA producing enzyme L-glutamic acid decarboxylase (GAD)

Honokiol, a Putative Anxiolytic Agent Extracted from Magnolia Bark, has No Diazepam-Like Side-Effects in Mice

Kuribara H, et al. J Pharm Pharmacol. 1999 Jan;51(1):97-103 Results suggest that honokiol is less likely than

diazepam to induce physical dependence, central depression and amnesia at doses eliciting the anxiolytic effect. It is also considered that honokiol might have no therapeutic effect in the treatment of convulsion

Effect of Honokiol on Activity of GAD(65) and GAD(67) in the Cortex and Hippocampus of Mice

Ku TH, et al. Phytomedicine. 2011 Oct 15;18(13):1126-9 Mice treated with daily injection for 7 days of honokiol

caused anxiolytic action which was similar to that was induced by 7 daily injection of diazepam in a elevated plus-maze test. In addition, the activity of hippocampal glutamic acid decarboxylase GAD(65) of honokiol treated mice was significantly increased than that of the vehicle or diazepam treated groups. These data suggest that honokiol causes diazepam-like anxiolytic action, which may be mediated by altering the synthesis of GABA in the brain of mice

HONOKIOL ANTIMICROBIAL ACTIVITY

Antimicrobial Activity of Magnolol and Honokiol against Periodontopathic Microorganisms

Chang B, et al. Planta Med. 1998 May;64(4):367-9 Honokiol has been used to treat such

periodontopathic microorganisms as Porphyromonas gingivalisPrevotella gingivalisActinobacillus actinomycetemcomitansCapnocytophaga gingivalisVeillonella disperNo cytotoxicity against human gingival fibroblasts and

epithelial cells; along with other gram-positive bacteria and fungi

Antimicrobial and Cytotoxic Activities of Neolignans from Magnolia officinalis

Syu WJ, et al. Chem Biodivers 1: 530–537, 2004 Honokiol has been used to inhibit the growth of

vancomycin-resistant enterococci (VRE) and methicillin-resistant Staphylococcus aureus (MRSA), in a dose-and time-dependent manner

HOW DO I USE HONOKIOL?

Honokiol in Cancer Treatment

Anti-Cancer Effects

Enhancement

Protectant

Combination and Synergy

Chemotherapy Cisplatin, Doxorubicin, Imatinib, Chlorambucil,

Gemcitabine, Adriamycin Heat Therapy Radiation Therapy (Honokiol enhances the sensitivity

of cancer stem cells to ionizing radiation) Honokiol in combination with radiation targets notch

signaling to inhibit colon cancer stem cells. Ponnurangam S, et al. Mol Cancer Ther. 2012 Apr;11(4):963-72

Metabolism

Honokiol Dosages

Active Cancer: Build up to 1 g X 3/day starting with 250 mg x 3 times a day

Prevention and Post Therapy: 1 g/day

Anti-inflammatory and Circulation Support: 250 mg x 2/day to 500 mg x 2/day

Periodontal disease: 500 mg x 2/day until condition improves, then 250 mg x 2/day

Antioxidant: 250 mg x 2/day

Anxiety: 250 mg x 2/day

Sleep: 250 mg before bed

Side Effects & Tolerance

No appreciable side effects Usually well tolerated Side effect (dose dependent)

Diarrhea Possible stomach discomfort

Reversed with warming herbs ginger, cardamom, etc.

Summary

Honokiol has broad antitumor activity Exhibits a desirable spectrum of bioavailability Crosses the blood–brain barrier Found also to have antioxidant, immune, anti-

inflammatory, neuroprotective, anxiolytic, and antimicrobial action

Honokiol is an anti-stress agent and a potent suppressor of oxidative damage and cancer

No appreciable toxicity Now available as a purified 98% honokiol extract