HORMONES: STEROIDS

Previous hormones: the prostaglandins and adrenalin.Insulin (a 51 amino acid protein) is another. A wide group of hormones contain the steroid structure.

H

HH

H12

34

5

67

8

9

10

1112

1314

15

1617

18

19

20 21

numbering shape

13

19

18

11 17

Steroid sources:

HO

O

O

HO

OH

Glu-Gal-OOH

O

O

Cholesterol Diosgenin (yams) Digitalis (fox glove)[from animals] [from plants]liver uses as source Drug companies use as source

Cholesterol found in all animals – liver converts to other steroids and can synthesize it if not enough (~200mg) in diet Isolated 1812 - structure determined 1932

Natural Sex Hormones:

OH

O

COCH3

O

OH

HO

O

Male Female Female[androgen] [gestogen] [estrogen]Testosterone Progesterone Estradiol

[Estrone]Secreted by testes ovaries ovariesControls 2o sex characs Prepares uterus for Controls 2o sex chs (hair, voice, muscles, etc) pregnancy (breasts, hair, menstrual

cycle)

T = 17b-hydroxyandrost-4-en-3-one AndrostaneP = pregn-4-ene-3,20-dione PregnaneE = estra-1,3,5(10)-triene-3,17b-diol Estrane

T P E

FEMALE CYCLE:

1) PITUITARY gland releases follicle stimulating hormone (FSH) which causes the follicle to develop in to an immature egg.

As the follicle develops, it releases ESTROGENS, which SHUT OFF FSH production so that only one egg develops at a time. These also cause expansion of the uterus lining.

2) ca. Day 14: ovulation occurs as egg leaves follicle – ESTROGEN production SHUT OFF but PROGESTERONE production STARTS

PROGESTERONE: keeps FSH shut off, keeps uterus developing, inhibits uterine contractions and enlarges breasts

3) IF egg fertilized: P produced throughout pregnancyIF egg NOT fertilized: P shuts off, FSH production begins,

uterus shrinks, egg flushed from body at ca. Day 28

Early in cycle: ESTROGENS presentLater in cycle: PROGESTINS present

So if we were to supply P or E: FSH stays shut off = no eggs = no pregnancy

However both are metabolised, so would need to inject

Oral Substitutes: ‘The Pill’ > $3 B/y

To make a pill we need:an active compound that could be oral and a cheap source

History:

OH

OEthisterone

COCH3

O19-norprogesterone

O

O

HODiosgenin

1934 H.H. Inhoffen: made Ethisterone as an oral progesterone substitute, but dose needed was large and no good source of material (1940: progesterone was $200/g)

1944 Russel Marker (Penn. State) establishes SYNTEX and finds a route to convert DIOSGENIN (from Mexican yams) to progesterone. Reduced progesterone to 20c/gram (current)

OH

OEthisterone

COCH3

O19-norprogesterone

O

O

HODiosgenin

1951: Carl Djerassi makes 19-norprogesterone; 4-8 x more potent than progesterone

1956: Syntex (cheap source) combines Djerassi and Inhoffen’s results to make Norethindrone (left)

Searle makes isomeric Norethynodrel (right)

1956-59 Pills are tested in S America

OH

O

OH

O

1960: the pill is released in N America and EuropeSEARLE: ENOVID Norethynodrel + MestranolORTHO: ORTHONOVUM Norlutin + MestranolSYNTEX: NORINYL Norlutin + Mestranol

The ingredients: Progestins (progesterone substitutes)

OH

O

OH

O

OH

O

Norethindrone Norethynodrel Norgestrel (Norlutin) (levonorgestrel)

(more recent variety)

Estrogen substitutes

OH

HO

OH

CH3O

Ethynylestradiol Mestranol

In 1960: pills used 10mg progestin + 0.15mg estrogen

By 1970: 1mg progestin + 0.1 mg estrogen

Today: even less estrogen

Ortho 10/11 uses:

0.5 mg norethindrone and 0.035mg mestranol for first 10 days,WHITE

1.0 mg norethindrone and 0.035mg mestranol for next 11 days, PEACH

Nothing for last 7 days GREEN

The estrogen prevents spotting (bleeding)

The blank week allows a normal period to occur

0.5, 0.75 and 1mg of the progestin (norethindrone) each with 35g of the estrogen (ethynylestradiol) + 7 blanks (green)

Ortho 7/7/7 is nowmore common, usesnorethindrone and ethynylestradiol

$72/3month

There are many manufacturers and many varieties:

those with 21 active pills allow period, blank pills always different color

If you take 28 active pills – no period

Also note: there are equivalent ‘Patch’ systems that slowly leak the same drugs (eg. EVRA (Ortho))

Seasonale (Dura-Med/Barr USA)(Palladin CAN)91 day regime, 84 active pills = one period every 3 months

Approved US (2003) and Canada (2007)Levonorgestrel (150g) + ethynylestradiol (30g)

Alesse (Wyerth)Even lower dosage of estrogen (20g): 21 (pink) or 28 day (pink, green) regimeLevonorgestrel (100g) + ethynylestradiol (20g)

Longer term solutions:

LYBREL approved June 2006 by FDA90 g levonorgestrel, 20 g ethynyl estradiol

Continuous use – no periods at all, though ~ 40%find some spotting during first year

WYETH REPORTS THATThe study showed that 99 percent of 187 participants experienced either a return to menses or became pregnant within 90 days after stopping the study drug.  Investigators reported that four women became pregnant before returning to menses and two women reported a return to menses more than 90 days after the completion of the study. Brings up the question: How to recognize if you are pregnant, while using? – Test

Will be sold as Anya in Canada – but still not approved?

http://www.lybrel.com/

DIANE-35 (Berlex Canada) (Schering US) $70/3 month

Side effects of birth control pills: better hormone level control, acne is cleared up

Diane-35 is specifically for severe acne control and excess body hair (it is an anti-androgen)

Studies indicate 4x increase in stroke risk

It contains cyptoterone acetate (2mg)(blocks androgen receptors) and ethynyl estradiol (35g)

OH

O

O

Cl

cyproterone

MINIPILLS (No Estrogen): Low dose of progestin every day egs. ‘Micronor’ (Ortho) and ‘Ovrette’ (Wyeth)

Ovulation normal: cervix secretes glue that prevents passage of sperm, and uterus is more hostile to implantation of an egg

No estrogen = less risk of blood clot 3x risk of pregnancy over combination pills (actual risk about 3%)

Long life injection - Depo-provera – (Pfizer) medroxyprogesterone acetate

COCH3

O

OCOCH3

More soluble in fat: slowly leached out150 mg injected every 3 months ($48)

If within 5 days of period, effective immediately

Success rate 99.7%

NOV 2004: Health-Canada and FDA alertshttp://www.hc-sc.gc.ca/hpfb-dgpsa/tpd-dpt/depo-provera_hpc_e.html http://www.fda.gov/bbs/topics/ANSWERS/2004/ANS01325.html

Warn about possible bone density loss (~3% in hip/y, 6% over 2 yrs) with prolonged use – seems to recover after stopping drug

IUD’s Intra uterine devices

Progestasert (a plastic T), leaks progesterone daily, lasts about 1 yr same problems of all IUD’s, see later

Mirena (Berlex): Plastic IUD with 52 mg of levonorgestrelleaks 20g/day, >99% effective, lasts about 5 years

Nuvaring (Organon): Approved 2004 (Canada) flexible transparent ringuses etonorgestrel (an 11-exomethylene norgestrel derivative) + ethynyl estradiol, so leaks P + E directly into the uterus

Copper IUD’segs. Nova-T and Gyne-T

Not much used in Canada: Problems: blood infections caused by irritation/perforation of uterus

Copper inhibits egg implantation Many lawsuits in 70's and 80's over the copper-7 and the Dalcon shield (~10M total users) - perforations followed by infections.For interesting account see: http://www.manhattan-institute.org/html/cjm_36.htm

MORNING AFTER PILLS

Until 1980 UVIC health services used DES (diethylstilbestrol, 25 mg)

Originally DES used in 40-50's to prevent miscarriages, but 15 years later, vaginal cancer in daughters observed, so this use stopped

Also used as cattle growth hormone (20% gain) - banned in 1973

As an abortant: no offspring to get cancer, but unpleasant drug to take

HO

OH

DES

NOW

4 OVRAL TABLETS (white not pink) (each has 500g of norgestrel (±) and 50g of ethynylestradiol)take 2 + 2 12 hrs later within 3 days of intercourse, > 98% effective

PLAN B (Paladin labs, Barr Labs (Can)) 2 tabs, 750g each levonorgestrel, taken within 12 hours of each otherOTC in Canada (as of June 06)

CONTRACEPTIVE SPONGESToday® Sponge (see Seinfeld episode!)

reserve for 1,000 mg of nonoxynol-9, an effective spermicidal agent that is continuously released into the vagina in small amounts through a 24-hour wear time Polyurethane foam traps and absorbs semen before sperm have a chance to enter the cervix: NOT VERY EFFECTIVE (84-87%)No protection against HIV/STD warning

Spermicide used on sponges is Nonoxynol-9:

C9H19 (OCH2CH2O)nOH

a non-ionic detergent used in shampoos

ABORTION PILL - RU-486 (Roussel-Uclaf, France)MIFEPREX (2000 in US)

Invented as cortisone antagonist, but progesterone antagonist as well

Approved for termination during first 49 days (63 d from last period)

OH

O

Me2N

Use 600mg (3 x 200mg), then oral prostaglandin (misoprostol) 48h later to induce contractions – abortion occurs within 4-22 h

96% success - must have surgical if fails.

Side effects: bleeding for ~9 days blood transfusion in 0.1% casesheart attack 0.02% however surgical abortion has risks at least as great or greater

Over last decade, >2M RU-486 abortions worldwide in 29 countries: >465,000 in US since 2000 – 4 deaths (from vaginal infection)

However, see also warnings at: http://www.fda.gov/cder/drug/infopage/mifepristone/default.htm

Not approved in Canada: one woman died in trial which was halted

Distributed by Danko Labs (NY), made in China.

RELATIVE EFFECTIVENESS OF VARIOUS METHODS (Average % PREGNANT DURING ONE YEAR OF USE)

BEST TYPICAL PREGS/MILMALE VASECTOMY 0.1% 0.15% 1,250FEMALE TUBES TIED 0.2% 0.4% 3,000NORGESTREL INSERTS 0.2% 0.2-0.4% 3,000DEPO OR PATCH 0.3% 0.3% 3,000ORAL COMB PILLS 0.5% 3% 12,000MINI PILLS 1.5% 3% 20,000COPPER IUD 0.8% 3% 20,000PROGESTIN IUD 2% 3% 25,000MALE CONDOM 2% 15% 75,000WITHDRAWAL 4% 18% 110,000DIAPHRAGM 6% 18% 120,000SPERMICIDAL SPONGE 6-9% 18-28% 170,000RHYTHM/TIMING 9% 23% 170,000NONE 85% 85% 850,000

Male Contraceptives: sperm take 3 months to make; about 108 released per day

Injected testosterone (and anabolic steroids) reduce sperm: probably shut down synthesis in testesmany undesirable side effects

Progestins and estrogens also reduce sperm production:reduce sexual interestcan lead to loss of bone and muscle mass

CHINA (1970-80, >10,000 subjects):

GOSSYPOL: seed of cotton plant daily oral pill gave low sperm count reversible in 3 monthsno reduction of performance, oral, cheap

BUT Side Effects: nausea, inability to sweat, takes 3 months to lower sperm levels, long term use causes sterility

HO

HO

CHO OH OH CHOOH

OH

India (2002):

RISUG (Reversible Inhibition of Sperm Under Guidance)Uses a polymer injected (or implanted) into vas deferens (sperm tube):effective for 5-10 yearsremovable by sodium bicarbonate washN. American trials since 2006 – not on sale yet?http://www.medicalnewstoday.com/medicalnews.php?newsid=40720

IMPOTENCE DRUGS - VIAGRA

~ 20-30M NA men suffer some kind of erectile disfunction

How it is supposed to work:

Muscles in the penis keep the artery walls contracted, restricting blood flow

To get an erection: during stimulation, nerves release NO which binds to the enzyme guanylate cyclase turning on synthesis of cyclic-GMP from GTP (guanosine triphosphate).

c-GMP causes Ca2+ ions to flow OUT of the penis muscles which then relax allowing blood flow in causing an erection

[c-AMP does same thing in the heart]

O

OHOH

Base'

OPOH

HOO

O

OH

Base'

OP

OHO

O

AMP/GMP c-AMP/c-GMP

N

N N

N

NH2

N

N

OH

H2N

Adenine

Guanine

At the end of the process, c-AMP and c-GMP are broken down by enzymes called PHOSPHODIESTERASES, PDE’s, muscles start contracting, erection dies

Inhibition of these enzymes would keep c-AMP/c-GMP around longer so a longer erection:

Viagra inhibits PDE-5 (penis)

For comparison:

Caffeine inhibits PDE-3 that breaks down c-AMP and so raises c-AMP levels:change in the flow of Ca2+ ions out of heart muscles (like adrenaline) causes ‘heart pounding’

cGMP = cyclic guanosine monophosphateGTP = guanosine triphosphateNANC = nonadrenergic-noncholinergic neuronsNO = nitric oxidePDE5 = phosphodiesterase type 5(from: www.viagra.com)

N

HNN

N

O2SN

N

EtO

O

PrN

NHN

N

O2SN

N

EtO

O

Pr

NH

N

O

O

O

O

H

sildenafil vardenafil tadalafil

VIAGRA (sildenafil) Pfizer: 50 or 100 mg blue pillsOnset ca. 20 mins to 1 h, lasts 4h (about 70% success rate)

Side effects: blue-green haze (poor PDE-5/PDE-6 selectivity)

Launched in March ‘98, by July ’98: 16 deaths

Not recommended for cardiac patients! 1999 sales: 1.22B$!

N

HNN

N

O2SN

N

EtO

O

PrN

NHN

N

O2SN

N

EtO

O

Pr

NH

N

O

O

O

O

H

sildenafil vardenafil tadalafil

Newer Versions

CIALIS (tadalafil) Lillyworks faster, lasts longer (24-36h) 10 and 20 mg, orange-yellow less blue-green vision

LEVITRA (vardenafil) Bayer/GlaxoSmithKlineorange, 10, 20 mg

ANABOLIC STEROIDS ($1.4B est /y in USA):

Steroids that build tissue (muscle mass) by increasing protein synthesis; also have androgenic properties (male sexual characteristics)

eg. Testosterone:builds musclesexcess shuts of FSH (decreases sperm production in males) leading to long term sterility, testicular atrophyacne, baldness

For women: facial hair, baldness, voice deepeningirreversible menstrual cycle changes

For youth: premature halting of growth (height)

O

OH

The liver oxidizes testosterone, so most on market are MORE FAT SOLUBLE (also more toxic to liver in general)

Stanazolol Androstenedione Primobolan [Ben Johnson] [Mark McGuire] [Alex Rodriguez]

Johnson lost 100m gold 1988 Seoul Olympicsstanazolol was a vet steroid, used to fatten cattle

McGuire broke home run record for St Louis Cardinalsadmitted to taking adrostenedione, not then banned by baseball

Rodriguez admits Feb ’09 to using Primobolan in 2003

OH

H

NHN

O

O

O

OH

H

OH

O

OH

O

OH

O

OH

O

Other ‘designer’ steroids:

Nandrolone THG=tetrahydro- Gestrinone Trenbolone gestrinone

[Barnes, Vezina] [Chambers, Toth]

Nandrolone: Barnes (‘96 Olympic shotput champ)now has lifetime ban after 2nd test in ‘98Vezina cost Canada a gold in ‘99 Pan-AmGames roller hockey

IOC limit = 2 ng/mL(men) and 5 ng/mL (women)for Nandrolone in urine (women are higher because produced as a metabolite from estrogens)

THG: June 2003 US Track championships: coach tipped the anti-doping agency about a new steroid (supplied a syringe - made identification easier) Used LC-MS to test for THG

Change in rules: samples now stored for future testing as new drugs are identified

Beijing Olympics 2008: >5000 drug tests, 9 positives

Athens Olympics 2004: >3000 drug tests, 24 positives

Is drug use on the decline…or are the athletes a step ahead?

Benzothiazepines:

Counteracts muscle fatigue by ‘stabilizing protein channels that leak Ca2+ from muscle cells during strenuous exercise’

In development, but some indications this may have been already used in Beijing (?)

Developers already have supplied WADA with a test that can now detect this in 0.1 ng/mL in urine or bloodSee: RSC Chemistry World Jan. 23, 2009

S

NO

S

NO

O

N

JTV519S107

Corticosteroids: steroidal hormones secreted in the adrenal cortex (immune and stress responses, inflammation, blood electrolyte levels, etc.)

Cortisol (11-OH) and cortisone (11-one)secreted daily (~ 25mg), most in morning, more if stressedprovides increased availability of energy from glucose, and is the bodies most potent inflammation reducer

COCH2OH

OHO

O

COCH2OH

OHO

OCortisone Prednisone (Deltasone)

Secreted by adrenal glands; oral pill as 5 mg dose or injected

Both reduce inflammation in doses > 25 mg/day: treat hivesAllergies and asthma – fluticasone propionate (ADVAIR)

Use must be tailed down slowly

Very severe side effects of overuse fat deposition (humps)protein (muscle) wasting Ca loss from bone (osteoporosis) water retention (conserves Na+) poor wound healing, ulcers, glaucoma

Cushing’s syndrome is ‘natural’ excess cortisol production (RU-486 used to treat)

Dexamethasone (Dexasone) is 25x more potent still, dose 0.5 mg

PROSCAR (finasteride) Merck

Dihydrotestosterone (DHT) is produced from testosterone by -reductase more as men age DHT stimulates (benign) growthof prostate – frequent urination

Proscar inhibits enzyme, about 50% of patents get shrinkage and relief (~400,000 prostate surgeries per year) At $1.40 per day = $1.5B per year!

Side effect of Proscar: reverses male pattern baldness!!

DANGER: pregnant women should not touch – deformation of the fetus is known to result!