WADA prohibited substances and their biochemical mechanisms 張振崗 國立臺灣體育運動大學 競技運動學系教授

M3 Gene doping The following, with the potential to enhance sport performance, are prohibited: The transfer of polymers of nucleic acids or nucleic acid analogues; 轉殖核酸聚合物或核酸類似物 DNA, RNA The use of normal or genetically modified cells. 使用正常或基因改造細胞。

S1 Anabolic agents 同化性物質 1. Anabolic Androgenic Steroids 2. Other anabolic agents

Anabolic Androgenic Steroids 同化性代謝雄性荷爾蒙

Anabolic Androgenic Steroids 同化性代謝雄性荷爾蒙 Testosterone 睪固酮 Gestrinone Nandrolone Danazol

androgen receptor modulators 選擇性 雄激素受體 調節劑 andarine ostarine BMS-564,929

25.12 Binding to the Response Element Is Activated by Ligand-Binding Ex Biochem c25-act transcript 25.12 Binding to the Response Element Is Activated by Ligand-Binding Binding of ligand to the C-terminal domain increases the affinity of the DNA-binding domain for its specific target site in DNA. Figure 25.19

S2 peptide hormones Erythropoietin (EPO) binding to erythropoietin receptor  highly glycosylated 醣化(40% of total molecular weight) half-life in blood ~ 5 hr, vary between endogenous and various recombinant 混合versions Additional glycosylation or other alterations of EPO via recombinant technology have led to the increase of EPO's stability in blood (thus requiring less frequent injections).

EPO binds to EpoR on red cell progenitor surface and activates Janus kinase 2 (JAK2) signaling cascade Jak-Stat Pathways. Jak-Stat pathways entail the receptor Janus kinase (Jak) and the signal transducer and activator of transcription (Stat).

EPO-related Methoxy polyethylene glyco-epoetin beta (CERA) 超長效紅血球生成素 Continuous erythropoietin receptor activator 連續性紅血球生成素受體活化劑  strucutre similar to the previous synthetic EPO drugs, except that it is connected to a chemical called polyethylene glycol (PEG), which makes it last 6-10 X longer in the body. Non-erythropoietic EPO-Receptor agonists asialo EPO 無唾液酸紅血球生成素

Hypoxia-inducible factors (HIF) Transcription factors Functions: angiogenesis 血管增生, development, metabolism HIF Stabilizers 穩定劑 HIF 活化劑: 氬氣及氙氣 氮氣

mechanism of HIF

Peptide hormones 人類絨毛膜性腺激素(Chorionic Gonadotrophin, CG), or human CG (hCG) 黃體化激素 (Luteinizing Hormone, LH) Both released during pregnancy懷孕 也可幫助女性懷孕 Manny Ramirez 相關釋放因子 releasing factors

CG and LH stimulate testosterone production Failed drug test if urinary testosterone to epitestosterone (T/E) > 6. hCG ↑endogenous production of testosterone and epitestosterone without increasing the urinary T/E ratio above normal hCG also be used following cessation of prolonged anabolic steroid use hasten a return to normal testicular function and size, when circulating testosterone very low.

生長激素 growth hormone 生長激素(GH)及相關釋放因子， 包括生長素釋素(GHRH) 其他生長因子 Insulin-like Growth Factor-1 (IGF-l) 似(類)胰島素生長因子

生長激素 growth hormone

S3 beta-2 agonists 致效劑 Beta-2 adrenergic receptor 腎上腺素受體agonists smooth muscle relaxation 平滑肌舒張(鬆弛) dilation of bronchial 支氣管 passages通道 vasodilation in muscle and liver  Used to treat asthma and other pulmonary disorders. anabolic effects (↑ muscle mass, ↓body fat percentage and faster recovery rates)? Most studies showed no ergogenic effect

S3 beta-2 agonists 致效劑

S4. Hormone and metabolic modulators 荷爾蒙及代謝調節劑 Aromatase inhibitors芳香環轉化酶抑制劑 Testosterone  estradiol androstenedione estrone

S4. Hormone and metabolic modulators 荷爾蒙及代謝調節劑 Selective estrogen receptor modulators (SERMs) 選擇性雌激素受體調節劑 anti-estrogenic substances 抗雌激素物質

S4. Hormone and metabolic modulators – 4. myostatin inhibitors Agents modifying myostatin 肌肉生長抑制素 function myostatin inhibitors 肌肉生長抑制素抑制劑 Disabled myostatin gene increase muscle size https://www.youtube.com/watch?v=yw4Ci6UrOK0

Myostatin mutations in animals Ex Biochem c9-genetics Myostatin mutations in animals

Ex Biochem c9-genetics

5. Metabolic modulators 代謝調節劑 5.1 腺苷單磷酸活化蛋白激酶調節劑 AMP-activated kinase, AMPK 例如AICAR 過氧化體增生劑活化受體delta致效劑 peroxisome proliferator-activated receptor delta, PPARdelta 例如 GW 1516

AMP-activated protein kinase activation Ex Biochem c5-CHO metabolism AMP-activated protein kinase activation

Effect of AICAR on AMPK Pokrywka, 2015

Pokrywka, 2015

Ex Biochem c25-act transcript PPARs pathway

GW501516, GW1516 PPARdelta agonist Ex Biochem c25-act transcript GW501516, GW1516 PPARdelta agonist Cause cancer in rats, development stopped by GSK

Effects of GW501516 on PPARdelta Pokrywka, 2015

Ex Biochem c25-act transcript WT: wild type TG: transgenic, 基因轉殖 expression of an activated form of PPARdelta in skeletal muscle Wang YX, PLOSB 2004

Ex Biochem c25-act transcript GW501516: PPARdelta agonist Wang YX, PLOSB 2004

Ex Biochem c25-act transcript GW1516 ↑endurance running performance after 5 weeks of training in mice V: vehicle only, GW: GW1516 Narkar, Cell 2008

Ex Biochem c25-act transcript GW1516 in combination with training↑oxidative enzyme expression in mouse quadriceps V: vehicle only, GW: GW1516, Tr: training, Tr+GW: training + GW1516 Narkar, Cell 2008

Proposed mechanism PGC-1alpha Ex Biochem c25-act transcript Proposed mechanism PGC-1alpha AICAR 5-aminoimidazole-4-carboxamide ribonucleotide Narkar, Cell 2008

Ex Biochem c25-act transcript 粒線體生合成的基因調控機制 Reznick et al, 2006

5. Metabolic modulators 代謝調節劑 5.2 胰島素 5.3 Trimetazidine 可代謝成amphetamine 安非他命 興奮劑

Ex Biochem c5-CHO metabolism

5. Metabolic modulators 5.3 Meldonium, Mildronate  anti-ischemia medication  on WADA list of substances starting from Jan 2016  Some athletes have been using it before it was banned, 2-17% previous urine samples ↓ inhibit carnitine synthesis ↓ fat oxidation (aerobic metabolism, requires oxygen) during ischemia ↑glycolysis, glucose metabolism Sport performance enhancement unclear

Mechanism of meldonium inhibit carnitine synthesis (-)

S5 Diuretics and masking agents 利尿劑與干擾劑 血漿擴張劑 甘油及靜脈注射albumin dextran, hydroxyethylstarch, 甘露醇 mannitol 利尿劑 Acetazolamide 一種利尿劑, inhibitor of carbonic anhydrase HCO3- + H+  H2CO3  H2O + CO2 In kidney, allowing the reabsorption of HCO3-, sodium, and chloride. By inhibiting this enzyme, these ions are excreted, along with excess water, 

S6 Stimulants 興奮劑 prohibited in-competition賽內禁用 Amfetamine (amphetamine): 安非他命 Cathine去甲偽麻黄鹼；mephedrone甲氧麻黄酮: 可轉換為安非他命 amphetamine Cathine mephedrone

S6 Stimulants 興奮劑 prohibited in-competition賽內禁用 Ephedrine 麻黃 Pseudoephedrine 偽麻黃鹼 一種「血管收縮劑」。過敏或感冒(流感)所引起的鼻塞，主要因鼻腔內微血管充血而腫脹的關係。此藥使鼻子內層的血管收縮，使充血腫脹的鼻腔恢復正常，而達到呼吸順暢的目的。可以紓解因為鼻竇腫脹而引起的不舒服﹑疼痛和呼吸不適。 ephedrine pseudoephedrine