1. E-2 Ethinyl Estradiol Joshua Nazeck Aquatic Toxicology Dr. Mason

2. Structure of the Compound and Physical and Chemical Properties Synthetic derivative of natural estrogen estradiol Synthetic derivative of natural estrogen estradiol High estrogen receptor potency High estrogen receptor potency Molecular formula Molecular formula C20H24O2 C20H24O2 Quickly absorbed when taken orally (most common form of administration) Quickly absorbed when taken orally (most common form of administration) Odorless Odorless Mol. Weight=296.41 g/mole Mol. Weight=296.41 g/mole

3. Physical and Chemical Properties continued Significance of the substitution at C17 with the estrane steroid. Significance of the substitution at C17 with the estrane steroid. Estradiol is quickly degraded by liver, substitution allows for resistance to this degradation Estradiol is quickly degraded by liver, substitution allows for resistance to this degradation Metabolized in the liver by means of Cytochrome P450 3A4 isoenzyme and CYP P450 2A6 Metabolized in the liver by means of Cytochrome P450 3A4 isoenzyme and CYP P450 2A6 Targets the estrogen receptor and Orphan nuclear receptor PXR Targets the estrogen receptor and Orphan nuclear receptor PXR variations occur in the absorption process and can have interactions with antibiotics variations occur in the absorption process and can have interactions with antibiotics Fully bound to plasma albumin Fully bound to plasma albumin Metabolized via hydroxilation of aromatic ring Metabolized via hydroxilation of aromatic ring Released into environment as a xenoestrogen via urine and feces. Released into environment as a xenoestrogen via urine and feces.

4. Uses and Application, Production History etc In almost all forms of modern combined oral contraceptives In almost all forms of modern combined oral contraceptives One of the most commonly used medications One of the most commonly used medications Synthesized by Hanz Herloff Inhoffen in 1938 Synthesized by Hanz Herloff Inhoffen in 1938 FDA approved the drug on June 25 th 1943 FDA approved the drug on June 25 th 1943 1 st marketed by Schering as Estinyl 1 st marketed by Schering as Estinyl FDA approval of estinyl withdrawn in 2004 due to discontinuation FDA approval of estinyl withdrawn in 2004 due to discontinuation Found under 56 different brand names and 46 different mixtures Found under 56 different brand names and 46 different mixtures

5. Applications continued Used for treatment of vasomotor symptoms associated with menopause, female hypogonadism, prostatic carcinoma-palliative therapy, breast cancer, and oral and emergency contraceptive Used for treatment of vasomotor symptoms associated with menopause, female hypogonadism, prostatic carcinoma-palliative therapy, breast cancer, and oral and emergency contraceptive

6. Mode of Entry in Aquatic Environment Urban runoff and wastewater effluent are the main sources of entry into the aquatic environment Urban runoff and wastewater effluent are the main sources of entry into the aquatic environment Enters system via human urine and fecal matter Enters system via human urine and fecal matter wastewater treatment facilities treat the water before dumping back into aquatic environment wastewater treatment facilities treat the water before dumping back into aquatic environment Not fully degraded and therefore large amounts still present in wastewater Not fully degraded and therefore large amounts still present in wastewater One of most commonly used medications we therefore would expect this to be found in large quantities at sites of wastewater runoff and highly developed areas. One of most commonly used medications we therefore would expect this to be found in large quantities at sites of wastewater runoff and highly developed areas.

7. Chemical reactivity with water, chemical speciation, physical half-life etc Found to be soluble very slightly soluble in water. Found to be soluble very slightly soluble in water. Experimental water solubility = 6.77 e-3 mg/L Experimental water solubility = 6.77 e-3 mg/L Physical half life 36 ± 13 hours. Physical half life 36 ± 13 hours. Has drug interactions with a large number of other drugs on the market Has drug interactions with a large number of other drugs on the market Too many drugs to note as well as varying interactions ranging from increasing effect and toxicity of theophylline and other toxins, to increased thrombotic risk due to estrogen Too many drugs to note as well as varying interactions ranging from increasing effect and toxicity of theophylline and other toxins, to increased thrombotic risk due to estrogen

8. Toxicity to aquatic life The toxicity of ethinyl estradiol is relatively minor according to the FDA website. The toxicity of ethinyl estradiol is relatively minor according to the FDA website. Mouse LD 50= 1737 mg/kg. Mouse LD 50= 1737 mg/kg. Symptoms include nausea vomiting and withdrawal bleeding in females Symptoms include nausea vomiting and withdrawal bleeding in females However studies have been conducted concerning the feminization of marine organisms recently on a large scale However studies have been conducted concerning the feminization of marine organisms recently on a large scale Up to 38% of Chinook Salmon found in the Sacramento and San Joaquin rivers were found to exhibit signs of feminization(http://www.ce.berkeley.edu/~sedlak/CALFEDwebsite.h tm#desbrow1998). Up to 38% of Chinook Salmon found in the Sacramento and San Joaquin rivers were found to exhibit signs of feminization(http://www.ce.berkeley.edu/~sedlak/CALFEDwebsite.h tm#desbrow1998). Ehtinyl Estradiol was found to be one of the many steroid contaminants in this river system Ehtinyl Estradiol was found to be one of the many steroid contaminants in this river system Feminization occurs for the most part during larval development Feminization occurs for the most part during larval development Impacts reproductive potential of marine organisms by essentially limiting the number of males present in a population. Impacts reproductive potential of marine organisms by essentially limiting the number of males present in a population.

9. Toxicity to aquatic life continued Potentially a very large problem Potentially a very large problem Ethinyl estradiol is most commonly found in sublethal concentrations Ethinyl estradiol is most commonly found in sublethal concentrations However even sublethal concentrations when present for multiple generations have negative effects. However even sublethal concentrations when present for multiple generations have negative effects. A study on the development of amphipods showed significant results concerning feminization and adverse effects associated with ethinyl estradiol concentrations A study on the development of amphipods showed significant results concerning feminization and adverse effects associated with ethinyl estradiol concentrations Hermaphroditism, disturbed maturation of germ cells, and disturbed spermatogenesis were found in all post F1 generation males exposed to EE Hermaphroditism, disturbed maturation of germ cells, and disturbed spermatogenesis were found in all post F1 generation males exposed to EE

10. Toxic effects noted Feminization of marine organisms Feminization of marine organisms Disturbed development and lowered fecundity Disturbed development and lowered fecundity Altered courtship behaviors in some fish (http://www.wef.org/NR/rdonlyres/F362B57D- 7F25-45AA-96D0- 66938800917A/0/MDWolfand.pdf) Altered courtship behaviors in some fish (http://www.wef.org/NR/rdonlyres/F362B57D- 7F25-45AA-96D0- 66938800917A/0/MDWolfand.pdf)http://www.wef.org/NR/rdonlyres/F362B57D- 7F25-45AA-96D0- 66938800917A/0/MDWolfand.pdfhttp://www.wef.org/NR/rdonlyres/F362B57D- 7F25-45AA-96D0- 66938800917A/0/MDWolfand.pdf LD 50=1737 mg/kg in mice LD 50=1737 mg/kg in mice Nausea, vomitting, and withdrawal bleeding in females Nausea, vomitting, and withdrawal bleeding in females

11. Mode of entry into organisms Non-aquatic organisms: usually enters as oral medication Non-aquatic organisms: usually enters as oral medication Aquatic organisms: Aquatic organisms: Absorption through gills or other rspiratory or absoprtive surface Absorption through gills or other rspiratory or absoprtive surface Found in wastewater effluent and urban runoff Found in wastewater effluent and urban runoff When this enters aquatic environment organisms are then subject to high levels of a steroid modified estrogen When this enters aquatic environment organisms are then subject to high levels of a steroid modified estrogen

12. Molecular mode of toxic interaction Ehtinyl estradiol is lipid soluble Ehtinyl estradiol is lipid soluble Diffuses easily through cell membrane Diffuses easily through cell membrane Diffuse into target cells and interact with protein receptor there. Diffuse into target cells and interact with protein receptor there. Target Cells: female reproductive tract, mammary gland, hypothalamus, and the pituitary gland Target Cells: female reproductive tract, mammary gland, hypothalamus, and the pituitary gland Then increase hepatic synthesis of sex hormone binding globulin, thyroid binding globulin and other serum proteins Then increase hepatic synthesis of sex hormone binding globulin, thyroid binding globulin and other serum proteins

13. Molecular Mode of Interaction Continued Suppresses follicle-stimulating hormone from the anterior pituitary Suppresses follicle-stimulating hormone from the anterior pituitary Combination of an estrogen with a progestin suppresses the hypothalamic-pituitary system Combination of an estrogen with a progestin suppresses the hypothalamic-pituitary system Decreases secretion of gonadotropin-releasing hormone Decreases secretion of gonadotropin-releasing hormone

14. Biochemical metabolism and breakdown Metabolized via Cyp P450 3A4 and Cyp P450 2A6 Metabolized via Cyp P450 3A4 and Cyp P450 2A6 Aromatic C ring broken down by hydroxilation Aromatic C ring broken down by hydroxilation After breakdown in the liver the estradiol bind to the estrogen receptors After breakdown in the liver the estradiol bind to the estrogen receptors Remaining ethinyl estradiol is excreted in the form of human waste Remaining ethinyl estradiol is excreted in the form of human waste

15. Works Cited 1- http://www.drugbank.ca/cgi-bin/getCard.cgi?CARD=DB00977.txt http://www.drugbank.ca/cgi-bin/getCard.cgi?CARD=DB00977.txt 2- Vandenburgh, Gurt et al. Effects of 17 -ethinylestradiol on sexual development of the amphipod Hyalella azteca. Ecotoxicolgy and Environmental Safety. Vol 54 Issue 2. 2003 216-222 3- Material Safety Data Sheet Ethinyl Estradiol. Science Lab.com, Updated 2005 4- Wolfland, Jordan. Active Ingredient in Oral Contraceptives (17-ethinylestradiol) Alters Male Competitive Courtship Behaviors and Secondary Sexual Characteristics in Fathead Minnows (Pimephales promelas). Journal of the US SJWP. Pgs 42-52. http://www.wef.org/NR/rdonlyres/F362B57D-7F25-45AA-96D0- 66938800917A/0/MDWolfand.pdf http://www.wef.org/NR/rdonlyres/F362B57D-7F25-45AA-96D0- 66938800917A/0/MDWolfand.pdf http://www.wef.org/NR/rdonlyres/F362B57D-7F25-45AA-96D0- 66938800917A/0/MDWolfand.pdf 5- Fergus, John. Benitez, J. Ethinylestradiol. INTOX-10 Meeting. ICPBS INCHEM 1997. http://www.inchem.org/documents/pims/pharm/pim221.htm#SectionTitle:5.5%20% 20Parenteral