"Muscle"Muscle Building"Building" NutritionalNutritional Supplements:Supplements: IsIs AndrostenedioneAndrostenedione AnAn AnabolicAnabolic Steroid?Steroid?

GregoryGregory A.A. Brown,Brown, MSMS

ExerciseExercise BiochemistryBiochemistry LaboratoryLaboratory Dept.Dept. ofof HealthHealth && HumanHuman PeformancePeformance TheThe IowaIowa StateState UniversityUniversity RogerRoger MarisMaris MarkMark McGwireMcGwire 6161 HomeHome RunsRuns inin aa SeasonSeason 7070 HomeHome RunsRuns inin aa SeasonSeason 19611961 19981998 Serum androgen levels after intake of 100 mg Androstenedione in Two Women 10 8 6 (nM) Androstenedione 4 2 0

24 20 16 Total Testosterone (nM) 12 8 4 0 0306090 (Mahesh(Mahesh && Greenblatt, Greenblatt, 1962)1962) Time After Ingestion (min) AndrogenAndrogen andand EstrogenEstrogen RelationsRelations D

B E DHEA-SDHEA-S DHEADHEA AndrostenedioneAndrostenedione EstroneEstrone (E(E11))

F G E

G G F

G DHTDHT TestosteroneTestosterone EstradiolEstradiol (E(E22))

H

33ββ-- 33αα-- AndrostanediolAndrostanediol AndrostanediolAndrostanediol EstriolEstriol (E(E33))

AA == PP 450450 SideSide ChainChain CleavageCleavage B= B= 33 ββ hydroxysteroidhydroxysteroid dehydrogenasedehydrogenase CC == 1717 αα hydroxylasehydroxylase D D == P450P450 17,17, 2020 lysaselysase EE == P450P450 aromatasearomatase F F == 1717 ββ hydroxysteroidhydroxysteroid oxidoreductase oxidoreductase GG == 55 αα reductasereductase H H == ketoreductaseketoreductase DoesDoes acuteacute androstenedioneandrostenedione intakeintake alteralter serumserum hormonehormone concentrationsconcentrations inin youngyoung men?men? * vs baseline (P<0.05) 40 * * * * * * * 35 * 30 * * 25 * PL ASD 20 Andro. 100 mg

15

10

Serum Androstenedione (nM) Androstenedione Serum 5 0

0 60 120 180 240 300 360 KingKing etet al,al, 19991999 Time After Intake (min) King et al, 1999 King King et al, 1999 King

Total Testosterone Free Testosterone (nM) (pM) 100 12 16 20 24 28 20 40 60 80 0 4 8 0

100 mg Andro. 010102030360 300 240 180 120 60 Time After Intake (min) ASD PL DoesDoes ChronicChronic AndrostenedioneAndrostenedione IntakeIntake EnhanceEnhance TheThe GainsGains InIn MuscleMuscle SizeSize andand StrengthStrength DuringDuring ResistanceResistance Training?Training?

KingKing etet al,al, 19991999 •• 2020 untraineduntrained subjectssubjects –– MaleMale (~23(~23 y)y) –– NoNo current/previouscurrent/previous supplementsupplement oror steroidsteroid useuse •• 88 weeksweeks fullfull bodybody resistanceresistance trainingtraining •• 33 days/weekdays/week •• 80-85%80-85% 11 RMRM •• Random,Random, doubledouble blindblind –– PLPL (rice(rice flour)flour) –– AndrostenedioneAndrostenedione (100(100 mgmg t.i.dt.i.d)) 1 RM Testing

Body Composition

Blood Chemistry

Sex Hormones

Muscle Biopsy

Supplementation

0 2468

KingKing etet al,al, 19991999 WEEKS 2500 Placebo Androstenedione * 2000 *

1500

1000

Lower Body Strength Lower Body 500

Knee Ex + R. Leg Flexion + L. Leg Flexion (N) Flexion Leg L. + Flexion Leg + R. Ex Knee 0 Before After Before After Training Training Training Training KingKing etet al,al, 19991999 King et al, 1999 King King et al, 1999 King Lean Body Mass (kg) 30 40 50 60 70 Training Before Placebo Training After * Training Before Androstenedione Training After * 7000 7000 * 6000 6000 *

5000 )

) 5000 2 2 m m µ µ µ µ µ µ 4000 µ µ 4000

3000 3000

Type I Area ( 2000 2000 Type II Area (

1000 1000

0 0 UN TR UN TR UN TR UN TR PL ASD PL ASD

KingKing etet al,al, 19991999 Placebo 35 Androstenedione 30 * * 25

20

15

10

Androstenedione (nM) Androstenedione 5

0 0 2468 KingKing etet al,al, 19991999 Weeks of Supplementation King et al, 1999 King King et al, 1999 King Total Free Testosterone (nM) Testosterone (pM) 100 120 140 160 12 16 36 20 40 60 80 20 24 28 32 4 8 0 0 0 Weeks of Supplementation 2468 Androstenedione Placebo IsIs AndrostenedioneAndrostenedione IntakeIntake Safe?Safe? King et al, 1999 King King et al, 1999 King HDL-Cholesterol (mg/dL) 48 46 24 26 28 30 32 34 36 38 40 42 44 Training Before Placebo Training After Training Androstenedione Before Training After * Placebo 170 * * Androstenedione 150 :pM) 1 130 110 90

Estrone (E Estrone 70 0.4 * * : nM) *

2 0.3

0.2

0.1

Estradiol (E 0.0 0.4 0.3 :nM) 3 0.2

0.1 Estriol (E 0.0 0 2468 KingKing etet al,al, 19991999 Weeks of Supplementation OtherOther NotableNotable FindingsFindings EffectEffect ofof 100100 mgmg androstenedione/dayandrostenedione/day onon MuscleMuscle ProteinProtein FractionalFractional SyntheticSynthetic RateRate 0.14

0.12 Control Androstenedione

0.10

0.08 %/hour 0.06

0.04

0.02

0.00 Pre Post Pre Post

RassmussenRassmussen et et al,al, 20002000 1800 1600 P=0.001 1400 1200 No Change 1000 800 600

Testosterone (ng/dL) Testosterone 400 200

Baseline Peak Baseline Peak 100 mg 300 mg Androstenedione Androstenedione LederLeder etet al,al, 20002000 EffectEffect ofof 100100 mgmg androstenedioneandrostenedione t.i.d.t.i.d. inin differentdifferent agesages

120 30 year olds 100 80 60 PL 40 ASD 120 40 year olds 100 80 60 40 120 50 year olds 100 Serum Free Testosterone (pM) Testosterone Free Serum 80 60 40 ‡ ****

Brown et al, 2000 01234 Brown et al, 2000 Week EffectEffect ofof 100100 mgmg androstenedioneandrostenedione t.i.d.t.i.d. inin differentdifferent agesages 5000 30 year olds 4000 3000 2000 1000 5000 40 year olds PL 4000 ASD 3000 2000 1000 5000 50 year olds 4000

Dihydrotestosterone (pM) Dihydrotestosterone 3000 2000 1000 ‡ **** 01234 BrownBrown etet al,al, 20002000 Week ToTo SummarizeSummarize

•• ChronicChronic androstenedioneandrostenedione intakeintake inin dosesdoses upup toto 100100 mgmg t.i.d.t.i.d. doesdoes notnot enhanceenhance •• TheThe increaseincrease inin musclemuscle massmass andand strengthstrength associatedassociated withwith resistanceresistance trainingtraining •• SerumSerum testosteronetestosterone •• MuscleMuscle proteinprotein synthesissynthesis ToTo SummarizeSummarize

•• ChronicChronic androstenedioneandrostenedione intakeintake inin dosesdoses upup toto 100100 mgmg t.i.d.t.i.d. doesdoes •• IncreaseIncrease SerumSerum DihydrotestosteroneDihydrotestosterone •• IncreaseIncrease SerumSerum EstrogensEstrogens •• DecreaseDecrease SerumSerum HDL-CHDL-C PossiblePossible AdverseAdverse HealthHealth ConsequencesConsequences

LoweredLowered HDL-CHDL-C •• IncreasedIncreased CVCV diseasedisease riskrisk byby 10-15%10-15%

ElevatedElevated EstrogensEstrogens HaveHave beenbeen linkedlinked to…to… •• IncreasedIncreased CVCV diseasedisease riskrisk •• PancreaticPancreatic CancerCancer •• GynecomastiaGynecomastia PossiblePossible AdverseAdverse HealthHealth ConsequencesConsequences

ElevatedElevated AndrostenedioneAndrostenedione maymay bebe associatedassociated with…with… •• ProstateProstate CancerCancer •• PancreaticPancreatic CancerCancer •• Neural/BehavioralNeural/Behavioral

ElevatedElevated DihydrotestosteroneDihydrotestosterone hashas beenbeen linkedlinked to…to… •• BenignBenign ProstateProstate HypertrophyHypertrophy •• BaldnessBaldness HerbalHerbal ExtractsExtracts

CompoundCompound Daily Daily DoseDose Purpose Purpose

SawSaw PalmettoPalmetto 480 480 mgmg Reduce Reduce DHTDHT metabolism/metabolism/ bindingbinding InhibitInhibit 55 αα reductasereductase

IndoleIndole-3-3-C-Carbinolarbinol 450 450 mgmg Inhibit Inhibit formationformation ofof estrogensestrogens

TribulusTribulus Terristris Terristris 1,350 1,350 mgmg Increase Increase LH,LH, FSH,FSH, TestosteroneTestosterone

γγ LinolenicLinolenic AcidAcid 1500 1500 mgmg Inhibit Inhibit 55 αα reductasereductase

ChrysinChrysin 300 300 mgmg Inhibit Inhibit aromatizationaromatization

BrownBrown etet al,al, 20002000 andand 20012001 AdditionAddition ofof TheseThese HerbalHerbal ExtractsExtracts

•• DoesDoes notnot preventprevent formationformation ofof estrogensestrogens fromfrom androstenedione,androstenedione, andand •• DoesDoes notnot increaseincrease serumserum testosteronetestosterone levels.levels.

BrownBrown etet al,al, 20002000 andand 20012001 FutureFuture DirectionsDirections

•• ToTo whatwhat extentextent dodo greatergreater (>300(>300 mg/d)mg/d) dosesdoses ofof androstenedioneandrostenedione affectaffect serumserum testosterone,testosterone, musclemuscle strengthstrength andand size?size? FutureFuture DirectionsDirections

•• WhatWhat areare thethe longlong termterm healthhealth consequencesconsequences ofof prolongedprolonged androstenedioneandrostenedione intake?intake? •• CardiovascularCardiovascular RiskRisk •• FertilityFertility FutureFuture DirectionsDirections

•• WhatWhat areare thethe effectseffects ofof ageage andand gendergender onon modifyingmodifying thethe responseresponse toto androstenedione?androstenedione?

•• WhatWhat areare thethe effectseffects ofof alternativealternative modesmodes ofof delivery?delivery? FutureFuture DirectionsDirections

•• WhatWhat areare thethe effectseffects ofof otherother androandro compounds?compounds?

•• WhatWhat areare thethe effectseffects ofof combiningcombining variousvarious formsforms ofof andro?andro? MatthewMatthew D.D. Vukovich,Vukovich, PhDPhD RickRick L.L. Sharp,Sharp, PhDPhD MarianMarian L.L. Kohut, Kohut, PhDPhD WarrenWarren D.D. Franke, Franke, PhDPhD TracyTracy A.A. Reifenrath,Reifenrath, M.S.M.S. KerryKerry A.A. Parsons,Parsons, M.S.M.S. DavidDavid A.A. Jackson,Jackson, M.S.M.S. N.L.N.L. Uhl,Uhl, B.S.B.S. EmilyEmily R.R. Martini,Martini, B.S.B.S. DouglasDouglas S.S. King,King, PhDPhD WeWe areare gratefulgrateful toto

ForFor theirtheir support. support.

ExtraExtra SummarySummary ofof OtherOther ClinicalClinical FindingsFindings • Chronic androstenedione intake in doses up to 100 mg t.i.d. does not alter serum concentrations of – Low Density Lipoprotein – Very Low Density Lipoprotein – Gamma-glutamyltransferase – Aspartate aminotransferase – Alanine aminotransferase – Globulins – Proteins – Prostate Specific Antigen

KingKing etet al,al, 19991999 andand BrownBrown etet al,al, 20002000 ExtraExtra SummarySummary

• A single 100-200 mg dose of androstenedione does not increase serum serum concentrations of testosterone in young men.

• A single 300 mg dose of androstenedione may slightly increase serum concentrations of testosterone ExtraExtra SummarySummary

• Chronic androstenedione intake in doses up to 100 mg t.i.d. does not enhance • Serum testosterone • Muscle protein synthesis • Adaptations to resistance training • lean mass • fat mass • muscle fiber size • muscle strength

KingKing etet al,al, 19991999 FSH mIU/ml) LH (mIU/ml) 0 1 2 3 4 5 6 0 1 2 3 4 5 6 7 0 Weeks of Supplementation 2468 Extra training data Extra training data ASD PL Extra training data 50 Extra training data

40

30 * * *

20 Placebo Serum HDL (mg/dL) HDL Serum Androstenedione 10

0 0 2468 Weeks of Supplementation 140 ExtraExtra trainingtraining datadata

120

100

80

60 PL 40 ASD Serum LDL (mg/dL) LDL Serum

20

0

0 2468 Weeks of Supplementation Maximal Muscle Strength (N) ExtraExtra trainingtraining datadata ASD (n=9) PL (n=10)

Wk 0 Wk 8 Wk0 Wk 8

Bench Press * 647 + 68 840 + 73 851 + 95 1,047 + 119

Biceps Curl * 287 + 19 331 + 22 351 + 21 391 + 25

Knee Extension * 717 + 46 1,024 + 57 770 + 45 1,095 + 52

Lat Pulldown * 657 + 25 787 + 44 672 + 35 876 + 50

Left Leg Flexion * 277 + 17 415 + 22 307 + 11 440 + 18

Right Leg Flexion* 292 + 14 430 + 28 311 + 10 445 + 16

Shoulder Press * 420 + 28 463 + 27 494 + 51 569 + 50

Triceps Extension * 311 + 24 390 + 19 329 + 31 454 + 30

Vertical Butterfly * 774 + 107 1,081 + 94 927 + 99 1,268 + 89

* Week 8 vs week 0 (main effect; P<0.05) ExtraExtra trainingtraining datadata Body Composition - * Wk 8 significantly different from Wk 0 (main effect, P<0.05) ASD (n=9) PL (n=10) Wk 0 Wk 8 Wk0 Wk 8 Height, cm 176.8 + 2.7 178.0 + 1.5 Body Mass, kg * 80.6 + 4.1 81.2 + 4.2 81.1 + 5.2 83.2 + 4.9 Lean Body Mass, kg * 61.2 + 2.5 64.1 + 2.4 63.1 + 2.6 66.0 + 2.5 Fat Mass, kg * 19.3 + 2.9 17.1 + 3.4 18.0 + 2.9 17.2 + 2.9 Body Fat, % * 23.5 + 2.4 20.3 + 3.0 21.3 + 1.9 19.9 + 2.1

Circumferences, cm Biceps * 31.9 + 0.8 32.4 + 0.8 32.6 + 1.4 33.4 + 1.3 Shoulder * 119.3 + 1.3 121.1 + 1.4 120.3 + 3.4 120.7 + 3.3 Chest * 99.6 + 2.0 99.2 + 1.9 98.0 + 3.8 100.9 + 3.4 Abdomen * 89.5 + 5.2 87.7 + 5.1 87.2 + 3.8 85.9 + 3.6 Waist * 86.6 + 4.2 83.7 + 3.6 84.9 + 3.9 83.7 + 3.7 Hips * 91.3 + 3.5 88.1 + 3.7 89.2 + 2.5 85.7 + 2.3 Gluteal * 101.7 + 3.1 98.2 + 3.1 100.4 + 2.5 97.4 + 2.3 Thigh 54.2 + 1.3 54.4 + 1.8 54.7 + 1.2 55.1 + 1.5 Calf 37.8 + 0.7 37.6 + 0.8 37.4 + 1.7 38.6 + 0.9 4400 * vs. 0, P<0.05 ExtraExtra AcuteAcute DataData 4000 * * 3600 * PL * * ASD 3200 * 2800 2400 2000 Serum DHT (pmol/L) DHT Serum 1600 1200

0 60 120 180 240 300 360 Time After Ingestion (min) KingKing etet al,al, unpublishedunpublished datadata 7 ExtraExtra AcuteAcute DataData 6 5 4 3 2

LH (mIU/ml) LH 1 PL 0 ASD 7 6 5 4 3

(FSH mIU/ml) (FSH 2 1 0 0 60 120 180 240 300 360 Time After Ingestion (min) Serum Total Testosterone (nmol/L) 10 12 14 16 18 20 22 24 2 0 4 6 8

010102030360 300 240 180 120 60 0 -100 mg ASD- Extra Acute Data Extra Acute Data Time After Ingestion (min) ASD ASD PL ASD ASD PL ExtraExtra SupportingSupporting AcuteAcute DataData 80 Androstenedione 70

60

50

40 Nmol/L 30

20 Testosterone 10

0

0 60 120 180 240 300 Minutes After Androstenedione Intake (100 mg)

RassmussenRassmussen et et al,al, 20002000 ExtraExtra 100100 mgmg androstenedioneandrostenedione t.i.d.t.i.d. inin differentdifferent agesages

40 30 year olds 30 20 10 40 40 year olds 30 20 10 40 50 year olds

Serum Androstenedione (nM) Androstenedione Serum 30 20 10 ‡ **** BrownBrown etet al,al, 20002000 01234 Week ExtraExtra 100100 mgmg androstenedioneandrostenedione t.i.d.t.i.d. inin differentdifferent agesages

30 30 year olds 20

10

0 30 40 year olds

20

10

0 30 50 year olds

20 Serum Total Testosterone (nM) Testosterone Total Serum 10

0

Brown et al, 2000 01234 Brown et al, 2000 Week ExtraExtra 100100 mgmg androstenedioneandrostenedione t.i.d.t.i.d. inin differentdifferent agesages

0.45 30 year olds 0.35 0.25 0.15 0.45 40 year olds PL 0.35 ASD 0.25 0.15

Estradiol (nM) Estradiol 0.45 50 year olds 0.35 0.25 0.15 **** Brown et al, 2000 01234 Brown et al, 2000 Week ExtraExtra 100100 mgmg androstenedioneandrostenedione t.i.d.t.i.d. inin differentdifferent agesages 4

3

2

1 PL (n=13 44 y) ASD (n=13; 41 y) Prostate Specific Antigen (ng/ml) Antigen Specific Prostate 0

01234 BrownBrown etet al,al, 20002000 Weeks of Supplementation ExtraExtra 20mg20mg sublingualsublingual cyclodextrincyclodextrin androstenediolandrostenediol administrationadministration 30 * * 25 * * *

20

15

10 SL-Androstenediol 5 Placebo

Serum Androstenedione (nmol/L) Androstenedione Serum 0 -20 0 30 60 90 120 150 180 BrownBrown etet al,al, 20022002 Time (min) Extra 20mg sublingual cyclodextrin androstenediol Extra 20mg sublingual cyclodextrin Extra 20mg sublingual cyclodextrin androstenediol Extra 20mg sublingual cyclodextrin Brown et al, 2002 Brown Brown et al, 2002 Brown Serum Estradiol (nmol/L) 0.06 0.08 0.10 0.12 0.14 0.16 0.18 2 06 01010180 150 120 90 60 30 0 -20 * administration administration * Time (min) Placebo SL-Androstenediol * * * * ExtraExtra 20mg20mg sublingualsublingual cyclodextrincyclodextrin androstenediolandrostenediol 200 administrationadministration * * 180 * 160 * 140 * * 120 100 80 60 40 SL-Androstenediol Placebo Free Testosterone (pmol/L) Testosterone Free 20 0 -20 0 30 60 90 120 150 180 BrownBrown etet al,al, 20022002 Time (min) MichaelisMichaelis Constants Constants forfor EnzymesEnzymes CatalyzingCatalyzing ConversionConversion ofof AndrostenedioneAndrostenedione toto OtherOther SteroidsSteroids

AndrostenedioneAndrostenedione

165165 nM nM 2525 nM nM 15001500 nMnM

DHTDHT TestosteroneTestosterone EstroneEstrone (E2)(E2)

55 ααReductaseReductase 1717 ββ-- Hydroxysteroid Hydroxysteroid DHDH AromataseAromatase A:T Conversions by Testicular Microsomes 200

17 ß-HSD Activity (pmol/h/mg)

100

0 T
A A
T

Blomquist,Blomquist, 19951995 Evidence that Oral Androstenedione Increases Serum Testosterone

Authors Subjects Dose [Testosterone ]

Mahesh & 2 Women 100 mg ↑ 500% Greenblatt

Hacker & ??? ??? ↑ 237% Mattern

Earnest et al, 8 men, 24 y 200 mg ↑ 15-20% 1999 Evidence that Oral Androstenedione Does Not Increase Serum Testosterone

Authors Subjects Dose [Testosterone ] 100 mg King et al, ↔ 10 men, 23 y 300 mg/day 1999 ↔ for 8 wk Rassmussen 6 men 100 mg ↔ et al, 1999 1 wk: 100 ↓ 26% Spratt et al, mg/d 6 men ↔ 1999 1 wk: 200 mg/d Ziegenfuss et 7 men, 28 y 100 mg ↔ al, 1998 Effect of 100- and 200 mg Androstenedione in 6 Healthy me 6

5 Testosterone (ng/mL) 4 *

3 100 mg 200 mg

Baseline Day 1 Day 7 Day 1 Day 7 Spratt et al, The Endocrine Society, 1999.

Androgenic 17 β-HSD Activity In Human Tissue

Adipose T Formation Adrenal A Formation Breast Endometrium Liver Muscle Myometrium Ovary Placenta Prostate Skin Spleen Testis

0.01 0.1 1 10 100 1000 pmol/mg protein/min