Standardized Phytoestrogen

Transcription

1 New Proprietary Standardized Phytoestrogen EstroG-100 Naturalendo Tech Co., Ltd. -1-

2 Changes in Secretion Estradiol Estron Before Menopause Peak Conc Lowest Conc Ratio 200~400 pg/ml 170~200 pg/ml 40~60 pg/ml Estradiol / Estron > 1 After Menopause Avg. Conc 5~20 pg/ml 30~70 pg/ml Ratio Estradiol / Estron < 1 Androstenedione converted estrone is major estrogen The conversion is made in fat tissue the more fat, the more estrogen Estron (E 1 ) Estradiol (E 2 ) Estriol (E 3 ) Odonnell MB. J Clin Pharmacol 1995;35(suppl):18S-24S Anderson F. Int J Fertil 1993;38(suppl 1):

3 Menopause Classification Premature Menopause About 8% Women (US) stops having period before age 40 Same symptoms as Menopause Peri-menopause Average women go through this phase age 45~49 Wildly fluctuating Estrogen Level Same symptoms as Menopause Menopause Estradiol level<50 pg/ml; FSH>50 miu/mg; no period for >1 yr* Average age for onset of menopause is 52 in US Symptoms last 2~8 years, many more than 5 years Surgical Menopause After Hysterectomy/Bilateral surgery * Definition: Dr. Joel Harglove, MD, Chairman, Vanderbilt Menopause Center, Nashville, TN -3-

4 Menopause Symptoms Physiological /Psychological Changes during Menopause Hot flashes, Night sweats Vaginal dryness & thinning of vaginal wall Inelastic skin and dry eye, Insomnia, nervousness, melancholia,,p paresthesia, vertigo(dizzy), fatigue, rheumatic pain, pounding of heart and headaches Long Term Post-Menopausal Health Risks Cardiovascular Diseases (CVD s) Osteoporosis -4-

5 HRT Women s Health Initiative Study (WHI) Strokes (44%), Blood Clots (100%), Alzheimer's (100%), CVD (29%), Breast Cancer (26%) FDA required Black Box Warning JAMA July 2002 Prescribed ONLY for <4 weeks -5-

6 Truth of Pomegranate ザクロを使った健康志向食品 - エストロゲンは本当に含まれているの? (Supplement using pomegranate Estrogen included?) 国民生活センター, [2000 年 4 月 6 日 : 公表 ] 結果 現状 今回調べた範囲ではエストロゲンは検出限界以下で検出されなかった また 延べ 7 個のザクロ果実を調べた結果でもエストロゲンが含有されていることの確認はできなかった またザクロの種子に含まれていた植物ホルモンのクメステロールも健康志向食品からは検出されなかった Any estrogen is not included under error range in this prove. In addition, any estrogen is proven not to be detected in 7 different pomegranate fruits. Coumestrol the phytoestrogen is not detected in the supplements either not as known to public to be included in the seeds. 独立行政法人国民生活センターの概要 ( 情報公開法第 22 条に規定する情報の提供 ) 国民生活の安定及び向上に寄与するため 総合的見地から 国民生活に関する情報の提供及び調査研究を行うこと -6-

7 Vulnerability of Black Cohosh 01/15/ /30/2007 The committee reviewed case reports suggesting a potential ti link between ingestion of the extracts and liver damage At the end of one year, there was no significant difference difference seen between the number of daily hot flashes and/or night sweats in any of the herbal groups compared to the placebo group, with an average of 0.6 less vasomotor symptoms per day in the herbal groups. There was a significant difference-4.06 fewer symptoms per day-in the hormone therapy group compared to the placebo groups -7-

8 Truth of Isoflavone Effective Dose As Isoflavone Aglycones, the Symptoms Improved w/ Daily Intake of 70 ~ 100mg, but Limitation of Dose Upper Limit of the Daily Intake of Isoflavone Aglycone is 30mg. Food Safety Commission (Japanese Government) Daily Intake Soy Isoflavone 34 mg Soy Isoflavone 36 mg Soy Isoflavone 60 mg Genistein 54 mg (Isoflavone 77mg) Genistein & Daidzein 70mg (Isoflavone 79mg) Isoflavone 100 mg Clinical Research on Isoflavones Researcher Washburn Penotti Research Result Published No effect 1999 No effect 2003 Duffy No effect 2003 Crisafulli Significant Improve 2004 Fauer Han Significant Improve Significant Improve

9 Is Isoflavone Carcinogenic?!! Genistein (a major isoflavone aglycone) May Cause Cancers ( NIH & FDA National Toxicology Program, Genistein Final, Jan, 2008) Exposure to Genistein i for 2 years caused mammary gland / pituitary gland adenoma in female rats. Exposure to Genistein for shorter duration following birth was also possibly associated with increased rates of pituitary gland and mammary gland tumors. -9-

10 Why Isoflavone Toxic? Cancer Causing Risks Isoflavone Could be Toxic because its high level of hormone-like action. It could be Hormone Dependent Cancer Causing Material due to its High Binding Affinity to Estrogen Receptor α, and β Relative Binding Affinity Estradiol Coumestrol Genistein 5 36 G Kuiper, Vol 138, No. 3 Endo 1997 α β Isoflavone from Sources other than Soybean Chinese Scholar Tree (Sophora p japonica): Isoflavone Contents 5 to 6 times Higher than Soybean Red clover has also Higher Isoflavone Contents than Soybean -10-

11 Science of New Herbal Ingredient Herbal extracts screened out of 71 herbal extracts via non- reproductive tract target tissue response (E-screen test) 3 herbal extracts were chosen: Cynanchum wilfordii, Phlomis umbrosa, and Angelica gigas Nakai Proven Safety About 400 years of documented use in Korea and China as folk medicine Registered as safe food ingredient in Korea Food Codex and in China No increase of uterus weight in ovariectomized rat tests Inhibition of proliferation of human breast cancer cell (MCF-7) No binding Affinity to both Estrogen Receptor α and β, cancer-inhibitory Safe: Acute & Multi-dose toxicity tests, Genetic toxicity tests Proven Efficacy in vitro, in vivo, and in 2 human group (Asian and non-asian) clinical studies -11-

12 MCF-7 cell & ER Binding Affinity MCF-7 proliferation inhibition Cynanchum wilfodii extract Phlomis umbrosa extract μg / ml 50 μg / ml 100 μg / ml 500 μg / ml 1000 μg / ml μg / ml 50 μg / ml 100 μg / ml 500 μg / ml 1000 μg / ml Estrogen receptor binding affinity test Absorbance (450 nm ) Affinity of EstroG to ER alpha E2 FGF Concentration (ug/ml) m) A bsorbance (450 n Affinity of EstroGto ERbeta 1.2 E FGF Concentration (ug/ml) EnBio Estrogen Receptor / Coactivator, Ligand Assay System -12-

13 Estrogenicity E-screen test: Screen herbal extracts for Estrogenicity Synergetic Effects of 3 Constituent herbal extracts confirmed -Leeat al.. Lab. Anim. Res. 24(2): (2008) Significantly improved in serum osteocalcin and FBMD in OVX rat No change in weight, liver, kidney, and uterus weight in OVX rat ht (g) terus and ovary weig U t Uterus weight Serum osteocalcin FBMD 1.4 * 1.2 * G1 G2 G3 G4 G5 G6 G7 G8 G9 Group L) Osteocalcin (ng/m * * * 4 * * G1 G2 G3 G4 G5 G6 G7 G8 G9 Group FBMD* (g / cm2 ) * * 0.7 -Kim at al.. Kor. J Food Sci. Technol. 40(3): (2008) * * * * * * G1 G2 G3 G4 G5 G6 G7 G8 G9 Group -13-

14 Toxicology 1. Single oral toxicity study No specific toxic symptoms in relation to test substance were observed. Minimal lethal dose (MLD) > 2,000 mg/kg. 2. Thirteen-week repeated (91 day) oral dose toxicity study of Estromon in Sprague-Dawley rats No observed adverse effect level (NOAEL) : upward 1,000 mg/kg in the male and female group 3. Genetic Toxicity Study Proven non-toxic Ames (Bacterial Reverse Mutation), Micronucleus, Chromosome Aberration -14-

15 Clinical study - Protocol Randomized Double-blind Placebo-controlled study (School of Medicine, Sungkyunkwan Univ. Samsung Cheil Hospital) ) Test material EstroG-200 Dosing period 12 months (May 2003-April 2004) Method Evaluation style Patients (n=47) Inclusion Criteria Double-blinded Long Term Safety Evaluation 23 subjects in placebo group & 24 active group Age>45 years old & Diagnosis of menopausal syndromes (average age=54) -15-

16 Clinical study I - Efficacy Climacteric Symptoms 5 times better improvement than placebo with significance Femoral Bone Mineral Density Significant improvement Serum hgh Level Significant improvement After 3 months OR=5.04 (95% C.I.= ) 18 1) Fisher s Exact Test After 12 months 0.746± ±0.13 (P<0.05) After 12 months 0.25± ±0.97 (ng/ml) (P<0.05) 05) Serum Osteocalcin Level Significant improvement After 12 months 6.02± ±3.01 (P<0.05) (ng/ml) Serum Alkaline Phosphatase Level Significant improvement Serum Triglyceride Level Significant improvement Lee DJ. J of Korean Society of Menopause. 11(1): (2005) After 12 months 73.35± ±14.87 (P<0.05) (IU/L) After 12 months 119.1± ±49.94 (mg/dl) (P<0.05) -16-

17 LA Times Ad for volunteers for trial -17-

18 Clinical study II (Non-Asian) Protocol Randomized Double-blind, Placebo-controlled study Test material Dosing gperiod Method Evaluation style Patients (n=61) Inclusion Criteria (CA, USA) EstroG months ( ) Double-blind Long Term Safety Evaluation 32 subjects in placebo group & 29 active group Age>42 years old & Diagnosis of menopausal syndromes (average age=53) -18-

19 Results Kupperman Menopause Index and Vaginal Dryness (Difficulties in Sexual Intercourse) Improved Significnatly Vaginal Dryness (Difficulties in Sexual Intercourse) Somatic Physiological - Hot flush/ Night sweat -Paresthesia - Vertigo - Fatigue - Muscular skeletal pain - Nervousness - Insomnia - Melancholia - Formication EstroG-100 was confirmed to improve both somatic and physiological symptoms with statistic significance. -19-

20 Vaginal Dryness (Difficulties in Sexual Intercourse) *; p-<0.05 compared between groups, **; p<0.01 compared between groups by t test ; p<0.01 compared between groups by Wilcoxon rank sum test #; p-<0.05 compared to baseline, ##; p<0.01 compared to baseline by paired t-test Vaginal Dryness Improved Unlike Black cohosh and Isoflavone -20-

21 Kupperman Index **;p<0.01 compared between groups by t test ; p<0.01 compared between groups by Wilcoxon rank sum test ##; p<0.01 compared to baseline by paired t-test Kupperman Index for Moderate or Severe e Menopausal Symptoms Improved -21-

22 Individual Menopausal Symptoms EstroG-100 (n=29) Placebo (n=32) Week 0 Week 6 Week 12 Week 0 Week 6 Week 12 Vasomotor 2.24± ±0.82 ** ## 0.79±0.73 ** ## 2.22± ±0.75 ## 2.06±0.76 Paresthesia 1.31± ±0.78 * ## 0.55±0.74 * ## 1.41± ±0.94 # 1.09±0.96 ## Insomnia 2.28± ±0.96 ** ## 0.97±0.82 ** ## 2.03± ±1.01 # 1.63±0.87 # Nervousness 1.72± ±0.69 ** ## 0.66±0.67 ** ## 1.56± ± ±0.75 Melancholia 1.93± ±0.68 ** ## 0.83±0.71 ** ## 1.59± ± ±0.74 Dizziness 0.97± ±0.49 ** ## 0.21±0.41 ** ## 0.75± ± ±0.80 Fatigue 2.21± ±0.77 ** ## 0.72±0.70 ** ## 2.00± ±0.90 # 1.59±0.80 # Arthritic Pain 1.59± ±0.94 ** ## 0.55±0.78 * ## 1.84± ± ±0.88 Headache 1.34± ±0.76 ## 0.66±0.77 ## 1.53± ±0.91 # 0.84±0.72 ## Palpitation 1.00± ±0.69 # 0.55±0.63 # 1.31± ±0.82 ## 0.75±0.84 ## Formication 0.83± ±0.44 * ## 0.28±0.45 ## 1.25± ±1.01 ## 0.72±0.96 ## * ; p-<0.05 compared between groups, ** ; p<0.01 compared between groups by t test ; p-<0.05 compared between groups, ; p<0.01 compared between groups by Wilcoxon rank sum test, # ; p-<0.05 compared to baseline, ## ; p<0.01 compared to baseline by paired t-test -22-

23 Clinical study - Safety Safety in both human clinical studies No serious adverse effects even without vaginal bleeding / spotting No change in body weight No significant changes in E2 and FSH No change in blood pressure, blood sugar, cholesterol, LDL/HDL -23-

24 Mechanism of Action Estrogenecity: In a non-reproductive tract target tissue response for e-screen assay, EstroG-100 promoted ALP synergetically more than any of the individual herbal extract Estrogen agonists and/or antagonists: to result in some benefits (eg, bone metabolism and menopausal symptoms) and not to influence human body to have adverse effects on endometrium and breast tissue based on the following available evidences: In two animal researches, EstroG-100 did not increase the uterus weight of ovariectomized rats while it increased femoral bone mineral density It did not show any affinity to both estrogen receptor alpha and beta Each herbal extract of EstroG-100 showed inhibitory effect of the proliferation of human breast cancer (MCF-7) cells In two clinical studies, it improved menopausal symptoms, bone density of femoral bone neck, bone markers without t any serious side effects with no increase of body weight and BMI and without influencing level of E2 and FSH -24-

25 Product Merit Award - NBJ -25-

26 KFDA HFFI Approval Health Claim - Safe since it is natural plant extract - Estrogen-like action confirmed since it increased bone density - Kupperman Index (as an instrument evaluating menopausal symptoms) improved - 10 individual symptoms such as hot flush, night sweat, paresthesia, sleep disorder, depression, vaginal dryness, etc. improved Technology and product selected by global l major industries i Awards and certificates -26-

27 NDI -27-

28 NHPD Licence NHPD Product License, Health Canada - Date of License: July 5, NPN(Natural Product Number): Helps to relieve the symptoms associated with menopause such as hot flashes, night sweats, paresthesia, insomena, nervousness, melancholia, vertigo, fatigue, rheumatic pain and vaginal dryness. -28-

29 Korean World-class Award Extract Compound of Cynanchum wilfordii Ministry of Knowledge & Economy

30 Presidential Citation Presidential Citation In recognition of the company s integrated contribution to the growth of the industry through edgestate technology development in agro-fishery food sector. Oct. 13, 2011 President Myung-bak Lee -30-

31 Media coverage Development and Globalization of Health Functional Food Ingredient for menopausal symptoms Korea Institute of Planning and Evaluation for Technology in Food, Agriculture, Forestry and Fisheries (President Byung-Rin Yoo) funded the R&D project titled Globalization of Proprietary Standardized Phytoestrogen by the Study of Active Ingredients and Clinical Effectiveness on Caucasians (Principal investigator PhD Bo-Yeon Kwak, Naturalendo Tech Co., Ltd) as one of development projects of agriculture and forestry technology. The total of 374 million Korean Won including a matching fund of 94 million Korean Won was funded from 2008 to 2010 for the project. The research team succeeded in developing a health functional food ingredient or HFFI called EstroG-100 TM containing root extract of Cynanchum wilfordii and Angelica gigas for improving the menopausal discomforts. In the clinical trial, EstroG-100 was proven to improve 10 of 12 typical symptoms such as hot flush, vaginal dryness, paresthesia, insomnia, nervousness, vertigo, melancholia, fatigue, rheumatic pain, and formication. EstroG-100 was confirmed to be safe as there was no changes in such variables as serum estrogen, follicle stimulating hormone, body weight, and bone density and as there was not reported to have any side effect in the study. -Proving efficacy and exporting of herbal extract (C. wilfordii, A. gigas)- -31-