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

I

N 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

H H

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

Nature Reviews Cancer 11, 85-95, 2011

Regulation of Cancer Cell Metabolism

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) %Migration of PC3 Cells

100 90 80 70 60 50 40 30 20 10 0 Control MCP 1 mg/ml MCP 2 mg/ml MCP 4 mg/ml %98 Honokiol 20um MCP 1 mg/ml + %98 Honokiol 20um

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

% Proliferation 120 100 80 60 40 20 0 24 hrs 48 hours 72 hours

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 hepatocytes  Human 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 B 6 ) 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 B 6 ) 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 gingivalis



Prevotella gingivalis



Actinobacillus actinomycetemcomitans



Capnocytophaga gingivalis



Veillonella disper

 No 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