Acta endocrinologic

Chief Editor

C H R I S T I A N H A M B U R G E R Copenhagen

Managing Editor

Κ. P E D E R S E N - B J E R G A A R D Copenhagen

J 0 R G E N S T A R U P Copenhagen

Associate Editor

Local Editors

J . - F . D y m l i n g , Sweden - C h r . Hamburger , Denmark

Β.-A. L a m b e r g , F i n l a n d - R. Nissen-Meyer , N o r w a y

G . P . v a n Rees, T h e Netherlands

J . T a m m , F e d e r a l Republic of G e r m a n y

A . W a l s e r , Switzerland

Consulting Editor

R u d i Borth, Toronto, C a n a d a

Volume 67 May-August 1971

P E R I O D I C A C O P E N H A G E N

G Ο Ν Τ Ε Ν Τ S

Authors Index, Vo l . 67 V I I Subject Index, V o l . 67 I X Advisory Panel August 1971 X X V

N U M B E R 1 · M A Y

Pliska V., Barth Τ. and Thorn Ν. Α.: Some metabolites of specifically t r i t i u m label led lysine-vasopressin. Ident i f i ca t i on by th in - layer chromatography and determination by the l i q u i d sc int i l la t ion technique 1

Pliska V., Thorn N. A. and Vilhardt H.: In vitro uptake and breakdown of t r i t i a t e d lysine-vasopressin by bovine neurohypophyseal and cortical tissue . . . . 12

Ortved Andersen 0., Bmnfeldt K. and Hansen Β. Α.: A n t i b o d y response i n the guinea p ig against iodinated p ig insul in preparations 23

Schelin U. and Lundin P. M.: A n electron microscopic study of normal and neo­plastic acidophil cells of the rat p i t u i t a r y 29

Petrusz P., Diczfalusy E. and Finney D. J . : Bioimmunoassay of g o n a d o t r o p i n s . 1. Theoretical considerations 40

Petrusz P., Diczfalusy E. and Finney D. ] . : Bioimmunoassay of gonadotrophins. 2. Practical aspects and tests of a d d i t i v i t y 47

Berle P. und Apostolakis M.: Pro lakt in-Konzentrat ionen i m menschlichen Plasma während Schwangerschaft u n d Wochenbett 63

Zaninovich Α. Α., Degrossi O. and Gotta H.: Effects of oestrogens on serum thyrox ine -b ind ing g lobul in capacity and on the per ipheral metabolism of thyroxine in patients w i t h hepatic cirrhosis 73

Lowy C, Wright A. D., Fräser T. R., Rubenstein A. FI. and Spitz I . : U r i n e excretion of insul in and growth hormone i n subjects w i t h renal fa i lure 85

Hedeland H., Dymling J . - F . and Η ok felt B.: Pharmacological i n h i b i t i o n of adrenaline secretion f o l l o w i n g insul in induced hypoglycaemia i n man. The effect of Catapresan 97

Damkjeer Nielsen M., Jorgensen Μ. and Giese J . : 1 2 5 I - l a b e l l i n g of angiotensin I and I I 104

Elana J . Ζλ. Ilardonk M. J . and Arenas Α.: Acute enzyme histocheinical changes in the zona glomerulosa of the rat adrenal cortex. I I I . The effect of pretreat-nient w i th deoxycorticosterone acetate on the changes seen alter peritoneal dialysis w i th 5 °/o glucose or after total nephrectomy 117

von Werder Κ., Sniilo R. P., Plane S. and For sham P. H.: P i t u i t a r y response to stress in Cushing's disease 127

Schildt Β. E. and Low FL: Relationship between trauma, plasma corticosterone and reticuloendothelial funct ion i n anaesthetized mice 141

Tamm J . und Voigt Κ. D.: The effect of metopirone on the u r i n a r y excretion of testosterone and ^tes tos terone i n normal males treated w i t h H C G or cypro-terone and in hirsute females 151

Peytremann Α., Veyrat R, and Midler A. F.: Variat ions i n plasma renin act iv i ty and ur inary aldosterone excretion i n normal subjects after salt restriction and acute p i tu i tary i n h i b i t i o n w i t h G-dehydro-16-methyIene hydrocortisone 159

Sundsfjord J . Α.: Radioimmunological determination of plasma renin act iv i ty during the menstrual cycle and d u r i n g acute progesterone adminis trat ion . . . . 174

Szamatowicz Μ., Drosdowsky Μ. and Jayle Μ. F.: The role of testosterone and androstencdione as precursors of f/;/testosterone i n guinea pigs (in vivo and in vitro studies) 187

Voute, J r . P. Α., van der Meer J . and Stangaard-Klooslerziel W.: Plasma renin act iv i ty i n W i l m s ' tumour 197

Kisch E. S.: D ispar i ty between the states of pseudopregnancy induced by reserpine and by cervical s t imulat ion i n the rat 203

N U M B E R 2 · J U N E

Miyai K., Amino N., Azukizawa M. and Kumahara Y.: In vitro effects of L A T S and T S H on phosphodiesterase act iv i ty i n human t h y r o i d homogenates 209

Kohler H., Haeberli Α., Binswanger C, von Gruenigen C. and Studer FL: E x ­perimental procedures leading to a relative accumulation of non- thyrog lobul in thyro ida l iodocompounds i n rat t h y r o i d glands 216

Spira 0., Gordon A. and Gross ] . : Fract ionat ion of 3-SS iodocompounds of the rat thyro id . I . Dialyzable and non-dialyzable iodocompounds 225

Spira 0., Gordon A. and Gross ] . : Fract ionat ion of 3-SS iodocompounds of the rat thyro id . I I . The presence and behaviour of a t h y r o i d a l iod ide-b inding peptide ( T I P ) 241

Petrusz P., Robyn C. and Diczfalusy E.: Ant igonadotrophic pro f i le of antisera against human gonadotrophin preparations. 1. Bio logical characterization of the antigens used for immunizat ion 249

Petrusz P., Robyn C. and Diczfalusy E.: Ant igonadotrophic pro f i le of antisera against human gonadotrophin preparations. 2. Biological characterization of the antisera 262

Nieschlag Ε., Wördehoff Β., Gilfrich FL J . , Michaelis J . und Overzier C: Kohlen­hydratstoffwechsel bei Hyperthyreose. Einfluss einer thyreostatischen Therapie auf orale und intravenöse Glucosetoleranz und Insulinaktivität 277

Karlberg B., Almqvist S. and Werner S.: Effects of synthetic p y r o g l u t a m y l -h is t idy l -pro l ine -amide on serum levels of t h y r o t r o p i n , Cortisol, g rowth hormone, insul in and P B I in normal subjects and patients w i t h p i t u i t a r y and t h y r o i d disorders 288

Moolenaar A. J . and van Seters A. P.: Gaschromatographic determination of steroids i n the urine of patients w i t h Cushing's syndrome 303

Jänne 0.: U r i n a r y excretion of mono- and disulphates of Cjq and Coi steroids w i t h reference to the menstrual cycle 316

Kaiser R. und Geiger W.: Die Östrogen- und Pregnandiolausschcidung bei H C G -Pseudograviditäten und nachfolgenden normalen Frühgraviditäten 331

Cupceancu B., Neiwiann F. und Steinbeck IL.: Beeinflussung der Ovar ia l funkt i on erwachsener weiblicher Ratten durch neonatale Behandlung m i t verschiedenen Steroiden mi t bekannter Gestagenwirkung 337

Lee A. E.: The l imitat ions of using mouse uterine weight as an assay of oestrogen antagonism 345

Holmdahl Τ. Η., Johansson Ε. D. Β. and Wide L . : The site of progesterone production i n early pregnancy 353

Nillius S. J . and Wide L . : Effects of progesterone on the serum levels of F S H and L H in postmenopausal women treated w i t h oestrogen 362

Goldzieher J . W. and Williams M. C: U r i n a r y testosterone glucuronide as a measure of endogenous testosterone production 371

Hansson V'., Tvcler Κ. J . and Altramadal Α.: Uptake and b i n d i n g of 5cc-dihydro-tcstosterone i n rat ventra l prostate in vivo 384

Tofi H.y Buns 0. and Nielsen E.: Vasopressin i n the diagnostic evaluation of p i t u i t a r y and hypothalamic funct ion 393

Persson L , Gyntelberg F., Heding L . G. and Boss-Nielsen J . : Pancrcatic-glucagon-l ike immunoreact iv i ty after intravenous i n s u l i n i n normals and chronic-pan­creatitis patients 401

Nieschlag Ε., Wombacher Η., Kroeger F. J . und Habighorst L . V.: Therapie eines mctastasierenden Inselzelltumors m i t Streptozotocin 405

N U M B E R 3 · J U L Y

Robyn C, UHermite M., Petrusz P. and Diczfalusy Ε.: Potency estimates of human g o n a d o t r o p i n s by a bioassay and three immunoassay methods 417

Matthies D. L . and Diczfalusy E.: Relationships between physico-chemical, i m m u ­nological and biological properties of human chorionic gonadotrophin. 1. Pro ­perties of human chorionic gonadotrophin as found i n tissues and body f luids . . 434

Matthies D. L . , Petrusz P. and Diczfalusy E.: Relationships between physico-chemical, immunological and biological properties of human chorionic gonado­t r o p h i n . 2. Biological and immunological behaviour of several species of human chorionic gonadotrophin 445

Warner S. M. and Freister S. D.: The early antibody response to human growth hormone 457

Ilipkin L . J . : The effect of c o r t i c o t r o p i n on the rat uterine weight response to human chorionic gonadotrophin 463

Woodings D. F. and Young P.: A rap id test of adrenocortical funct ion using Homact id (synthetic α, , 1 - 2 8 A G T H ) 470

Littmann K.-P., Gerdes FL und Winter G.: K ine t ik und Charakterisierung der östradiolsensitiven 17/?-Hydroxysteroiddehydrogenasen i n der menschlichen Leber 473

Sunds fjord ] . Α.: Plasma renin ac t iv i ty and aldosterone excretion d u r i n g pro ­longed progesterone administrat ion 483

Hobson B.: Immunoassay of human p i t u i t a r y lute in iz ing hormone ( L H ) i n urine. A comparison between a haemagglutination and radioimmunosorbent method . . 491

Kaalund Jensen I L and Blichert-Toft M.: Invest igation of p i tu i tary -adreno -cortical function in the elderly d u r i n g standardized operations and postopera­tive intravenous metyrapone test assessed by p l a s m a Cortisol , plasma compound S a n d eosinophil cell d e t e r m i n a t i o n s 495

Boss N., Kluge F., Müller Ο. Α., Pickardt C. R. and Scriba P. C: Quantitat ive assay of the suppressive effect of synthetic corticoids i n man. Evaluat ion of the c i r c a d i a n r h y t h m of s e r u m Cortisol a f ter s ing le o r a l doses of f luocorto lone a n d prednisolone 508

Wenzel Μ.: Oxydation des Anabol ikums l -Methy l -androst - l - en -17 / ? -o l -3 -on m i t Wasserstofftransfer auf Östron. In vitro Untersuchungen mi t Schnitten von Rattenorganen 517

Melville Kelly R. W. and Mitchell F. L . : Fai lure to detect Sß,ll^,17a,21-tetra-hydroxy-5-pregnen-20-one i n plasma obtained f rom human umbi l i ca l cords and human in fant urine 531

Simmons J. E.: Uptake of [ 1 , 2 - 3 H ] testosterone i n oes trogenized male rats 535 Steinbeck FL, Cupccancu B., Mehring Μ. and Neuman F.: Influence of neonatal

gestagen inject ion on the d i f f erent ia t ion of psychosexuality i n female rats . . . . 544

Ciaassen V., Μ or sink L . and de Wächter Λ. Μ.: Influence of dydrogestcrone ((>-dehydroretroprogesterone, Duphaston®) on ovulat ion i n the rat , rabbit and monkey 551

Plait D. and Frackenpohl I L : Pept idyl peptide hydrolases, cathepsin and col -lagenase (?) of the rat uterus dur ing postpartum invo lu t i on 563

Becker L L , Bene P., Wedle A. and Voigt Κ. D.: Influence of late gestation and early puerperium on l i p i d and carbohydrate metabolism i n normal females . . . . 570

Brandau Η. und Brandau L . : Enzymologische Studien am Arrhenoblastom 577 Moore H. C, Borvendeg ] . and Wilson K.: The effect of foetal removal on the

blood pressure i n steroid hypertensive rats 590 Bene P. und Aposlolakis M.: Inkubationsversuche m i t einem A n t i h u m a n - P r o l a k t i n 597 Wägar G.: Increase i n t h y r o i d uptake of radioiodine induced by act inomycin D 605 Siersba?k-Nielse?i K. and Molhohn Hansen ] . : The b ind ing capacity of thyrox ine -

b inding g lobul in i n cerebrospinal f l u i d 616

N U M B E R 4 • A U G U S T

Vingerhocds A. C. M., der Rinderen P. /., Thijssen ] . Η. H. and Scharwz F.: Detection of an ACTH-secre t ing bronchial carcinoid tumour eighteen months after adrenalectomy for Cushing's syndrome 625

Simons son B.: Uptake of 3 H - c o r t i s o l by rabbit polymorphonuclear leukocytes in vitro 634

Kakihana R., Butte J . C , Hathaway A. &: Noble E. P.: Adrenocort i ca l response to ethanol i n mice: M o d i f i c a t i o n by chronic ethanol consumption 653

Oh R. and Tamaoki B.: Occurrence of 19-oxoandrost-4-ene-3,17-dione in the course of oestrogen biosynthesis by equine testicular microsomes 665

Cohen S. L . : The excretion of »labile« oestrogens d u r i n g human pregnancy. I . N o r m a l pregnancy 677

Cohen S. L . : The excretion of »labile« oestrogens d u r i n g human pregnancy. I I . Diabetic pregnancy 687

Patwardhan V. V. and Lanthicr Α.: The in vitro f ormation of androstenedionc f rom Coj steroids by follicles isolated f r om human ovaries. A time study 698

Pal S. B.: The pattern of ur inary excretion of i n d i v i d u a l 17-ketosteroicls i n rhesus monkeys 711

Adhikary P. M. and Harkness R. Α.: The use of carbon skeleton chromatography for the detection of steroid drug metabolites: The metabolism of anabolic steroids in man 721

Bird C. Green R. N., Calandra R. S., Connolly J . G. and Clark A. F.: Kinetics of 3 H-testosterone metabolism i n patients w i t h carcinoma of the prostate: Effects of oestrogen administrat ion 733

Kowalewski K., Heron F. and Russell J . C: Effect of sex hormones on the meta­bolism of collagen and i 5 C a by the bones of young gonadectomized rats 740

Johnsen S. G..* Evaluat ion of g o n a d o t r o p i n analyses i n male hypogonadism . . 756 Quabbe H.-J., Helge Η. and Kubicki S.: Noc turna l growth hormone secretion:

Correlat ion w i t h sleeping E E G i n adults and pattern i n chi ldren and adolescents w i t h non -p i tu i ta ry dwarf ism, overgrowth and w i t h obesity 767

Ljunggren J.-G., Sedvall G., Levin K. and Fyrö Β.: Influence of the l i t h i u m i o n on the biosynthesis of thyroxine and thyrog lobul in i n the rat 784

Liewendahl K., Tölterman ] . and Lumberg Β-Α.: Determinat ion of total and free serum thyroxine i n t h y r o i d diseases 793

Λ C Τ Α Ε Ν D Ο C R I Ν Ο I . Ο ϋ I C Λ

67 (1971) 508-5 Κ)

I I . Medizinische K l i n i k der Universität München, Deutschland

Q U A N T I T A T I V E A S S A Y O F T H E

S U P P R E S S I V E E F F E C T O F S Y N T H E T I C C O R T I C O I D S I N M A N

Evaluation of the circadian rhythm of

serum Cortisol after single oral doses of fluocortolone

and prednisolone

By

N. Boss, F. Kluge, Ο. Λ. Müller, C. R. Pickardt

and P. C. Scriba''

A B S T R A C T

The suppressive effect of synthetic corticoids on the human hypothala -mic -p i tu i ta ry -adrena l axis was analyzed by determining the integral differences of the circadian serum Cortisol levels between control subjects ( N = r 25) and subjects receiving di f ferent single oral closes of p r e d n i ­solone ( N = 29) or fluocortolone ( N = 36). L inear log dose-response curves were obtained by this procedure w i t h the indices of precision for prednisolone of λ = 0.221 and for fluocortolone of λ = 0.149. W i t h this method i t is possible to compare the suppressive effect of di f ferent corticoids. The relative suppressive potency of corticoids should not be expressed by ratios or factors, but rather by log dose-response curves of the type: response = b · log dose + a.

T h i s study was conducted in order to develop a procedure for quantitating

the suppressive effect of single oral doses of synthetic corticoids on the h y p o -

t h a l a m i c - p i t u i t a r y - a d r e n a l axis in m a n , since the avai lab le data in the l i t e r a ­

ture were considered to be unsatisfactory (Hedner 1967; Radvila et al. 1969;

Hochheuser et al. 1969; Beihge 1970). Heclner (1967) administered o r a l l y four

synthetic corticoids at four equipotential dose levels at midnight a n d c o n ­

sidered the reduction in the 8.00 a. m. p l a s m a corticosteroid values as the

H e r r n Prof. D r . med. Η. Schwiegk zum 65. Geburtstag gewidmet. * Supported by the Deutsche Forschungsgemeinschaft (SFB 51).

508

suppressive effect. R e c t i l i n e a r log dose-response curves were obtained w i t h

indices of precision from A = 0.19 to / = 0.39. Radvila el al. (1969) followed

the c i r c a d i a n v a r i a t i o n of serum 11-hydroxycorticosteroids after ora l corticoid

administrat ion , but only single dose levels of dexamethasone and triamcinolone

were used.

T h e present study w a s performed in order to obtain l inear log dose-response

curves for prednisolone a n d fluocortolone w i t h reasonable indices of p r e ­

cision (λ). T h e r e f o r e var ious dose levels of these corticoids were administered

o r a l l y and the curves of the c i r c a d i a n serum C o r t i s o l values were subsequently

a n a l y z e d unti l the intersection w i t h the n o r m a l c i r c a d i a n C o r t i s o l profile. B y

this means, both the degree a n d durat ion of the suppression of the h y p o -

t h a l a m i c - p i t u i t a r y - a d r e n a l axis by a single corticoid dose could be taken as

response.

P r e l i m i n a r y results of this study have been reported (Boss et al. 1970: Scriba

et al. 1970a).

M A T E R I A L A N D M E T H O D S

The subjects under study were hospitalized for various non-acute, non-endocrine diseases. Subjects were either selected for the control group or suppressed w i t h only one single dose of the synthetic corticoids used, usually d u r i n g convalescence shortly before being discharged f r o m the hospital . The control group ( N = 25 males) had an average weight of 73.0 ± 10.6 kg and were 41.7 ± 16.1 years of age (mean ± S D ) . The prednisolone group ( N = 29 males) and the fluocortolone group ( N = 36 males) had average weights of respectively 75.2 ± 11.1 kg, and 70.1 ± 11.7 kg, and were respec­t ive ly 41.0 + 14.0 and 41.5 ± 15.5 years of age. The two to ta l ly adrenalectomized patients were both females (Μ. M . , 43 years; L I . M . , 46 years). Adrenalectomy i n these cases was performed several years before this study because of Cushing's syndrome w i t h bi lateral adrenal hyperplasia.

Prednisolone (Pregna-l,4-dien-llß,17a,21-triol-3,20-dione) and fluocortolone (6a-f luoro-16a-methyl-pregna-1.4-dien- l l /5 ,21-diol -3 ,20-dione) were given ora l ly as tablets (5 mg) at 7.00 a. m. i n doses indicated i n Figs. 3 and 4. Blood was collected in po ly -styrol tubes at the intervals indicated i n Figs. 1, 5 and 6; the serum was stored at - 2 0 ° C until determination.

Serum Cortisol was determined f luor imetr i ca l ly as previously described (Kluge et al. 1970; Scriba et al. 1970&). The suppressive effect of prednisolone and fluocortolone on the circadian r h y t h m of serum Cortisol was determined as shown i n Figs. 1 and 2.

Statistical analysis was performed as indicated throughout the text (Wcdlis 8c Roberts 1969; Gaddum 1953). The authors are indebted to D r . P. Kuhbier, München, for his valuable help i n the statistical evaluation.

R E S U L T S

T h e suppressive effect of various single oral doses of synthetic corticoids a d ­

ministered at 7.00 a. m. was determined by ca lculat ing the integral differences

509

^ g C O R T l S O L g o o + 6S? ( C O Ν Τ R O L G R O U Ρ )

1 0 0 m l - J 3± =F

A 6?° • 6°£ ( C O R T 1 C O I D S U B J E C T ) 7°° C O R T I C O I D A D M I N I S T R A T I O N

( 1 0 m g F L U O C O R T O L O N E )

Fig. 1 a and b. Example of the suppression of c ircadian serum Cortisol levels by a single ora l dose of corticoid, i . e. fluocortolone at 7.00 a. m. The circadian r h y t h m of serum Cortisol levels of the controls (mean ± S D , Ν = 25) for 48 h is represented by the upper curve in

both parts of F ig . 1 ( + ) .

a) Measured curve. The serum Cortisol levels measured i n a subject given 10 m g f luocortolone at 7.00 a .m . are shown as a n example by the lower curve (o). Since the values of the corticoid subject at 6.00 a. m. on the First and third day are different from the mean Values of the controls, each Cortisol value of the corticoid treated subjects, receiving f luocortolone

or prednisolone, had to be corrected.

b) Corrected curve. The corrections for the i n i t i a l 6.00 a. m. and for subsequent values were performed by m u l t i p l y i n g the measured Cortisol values of each corticoid treated subject (here fluocortolone) by a factor F, derived f r om d i v i d i n g the 6.00 a. m. values of the first and t h i r d day of the control group by the 6.00 a. m . values of the f irst and t h i r d day of each corticoid treated subject. Analogous corrections were then appl ied to the Cortisol levels of each corticoid treated subject receiving either f luocortolone or prednisolone. The corrected levels of serum Cortisol of the subject r e c e i v i n g 10 m g f luocortolone are shown i n the lower curve (o). The hatched area between the control and the corrected corticoid curve was calculated as the difference between the integrals of the two curves (A integral = suppressive effect) f r om the start at 6.00 a. m. u n t i l

the intersection (SP).

CORTISOL 1 0 0 m l

Fig. 2. Schematic presentation of the calculation of the suppressive effect of a single oral dose

of corticoid, i . e. f luocortolone or prednisolone.

SP

The difference of the integrals (A \ ) of the circadian Cortisol curves of the controls ο

( y N ) and of a corticoid subject (y) f r om the start (t = 0) u n t i l the intersection (SP) was calculated as the sum of the single areas F t to F n :

SP

A \ = F t + F 2 + + F I l _ 1 + F I l

F ,

( y N o - y J + ( y N i - y i )

M i - l ~ ( ν Ν η - 2 - ν ι ι - 2 ) + ( y N n - l ~Yn-i)

= ( y N n - i - y n - i ) · - T -

h was derived f r om geometric formulae. The calculation was performed w i t h a com­puter (O l ive t t i Programma 102). The authors are indebted to D r . Peter v. Bre i ten-

lohner, München, for setting up the programme for the computer.

511

b e t w e e n the c i r c a d i a n C o r t i s o l levels in the controls a n d in the subjects re ­

ceiving corticoids. T h e r e was a l inear log dose-response relationship between

the log dose of prednisolone, or of fluocortolone, a n d the suppressive effect SP

A \. T h i s was shown for the absolute dose of corticoid in m g ( F i g . 3) and also ο

for ßg per kg body weight ( F i g . 4 ) . T h e correlat ion coefficients (r) and the

indices of precision (λ) for both prednisolone a n d fluocortolone are satis­

factory (Figs . 3 a n d 4) , i n v i e w of the fact, that the model used might be

considered as a method of bioassay in the h u m a n subject .

Δ j p q C O R T l S O L . J J L 1 0 0 m t J

Λ

3 0 0

250

2 0 0

150

100

50

PREDNISOLONE ( · ) N * 2 9 b «86.9 α - 60 .6 r . * 0 . 6 A 3 , X - 0.221

® »meant SD

1 FLUOCORTOLONE(o ) ο % b = 112.0 ; a =10.9 ; N = 3 6

r = +0.762 ; 0 . K 9

1 m g Q39C 2 5 m g 5 m g

- l o g d o s e

10 m g

Fig. 3.

2 0 m g 3 0 m g 5 0 m g -

SP

Suppressive effect (A \) of prednisolone ( · ) and of f luocortolone (o) plotted as ο

log dose i n mg. Single values and means ± S D of the suppressive effect, calculated as described i n Fig. 2, are shown for the log dose of corticoids, given as a single dose at 7.00 a. m. Linear log dose-response curves were calculated f rom y = bx + a w i t h

Σ*Υ ~ N x y the regression: b = — ,

5 2 χ 2 - Ν χ 2 the suppression of the f ict ive dose of 1 m g :

a = y l o r χ = 0, v S y - N x y the correlation: r —·

l / ( 2 x 2 - Ν χ 2 ) " ( 2 ν 2 - N y 2 ) '

and the index of precision: λ = ; where s h

b The two dose-response curves intersect at I P .

_ s y ] / l ~ r 2

s x γ n - 2

512

/ W O R T I S O L . J J L 1 0 0 m l J

3 0 0

body weigh!

Fig. 4. SP

Suppressive effect (zl \) of prednisolone ( · ) and of f luocortolone (ο) plotted as ο log dose in / / g per kg body weight.

T h e l inear log dose-response curves were calculated as described i n F ig . 3.

A p p a r e n t l y the relat ive suppressive potencies of the two corticoids compared

cannot be character ized by a simple factor or ratio (Hedner 1967). Instead . SP

the suppressive effect has to be expressed by dose-response curves Δ \ = b · log ο

dose -f- a (F igs . 3 and 4). T h e two curves obtained for prednisolone a n d for

fluocortolone are sl ightly different w i t h regard to their regressions (b). T h e

dif ference between the regressions is h a r d l y of statistical s ignif icance w h e n

the dose is plotted ( F i g . 3) in mg (/^ < 0.15), or (Fig- 4) in tug per kg body

weight (P < 0.2).

H o w e v e r , the assumption that the two curves intersect at I P (Figs . 3 a n d 4)

is supported by the fact, that the values for a, w h i c h denote the suppression

at doses of respectively 1 mg, and 1 μξ per kg body weight, are s igni f icant ly

different ( P < 0 . 0 5 ; prednisolone: Ν = 29, f luocortolone: Ν = 36). T h e log

dose-response curves do not cross at the intersection of the ordinate a n d

abscissa, since the log of a dose of zero w o u l d be indefinite .

N o signif icant difference was found for the var ious corresponding groups

receiv ing prednisolone or fluocortolone i n doses r a n g i n g from 2.5 to 50 m g

(Figs . 3 a n d 4) . O n the other h a n d , a dose as low as 2.5 mg of prednisolone or

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of fluocortolone suppressed the C o r t i s o l levels s igni f icant ly as compared with

the controls (P < 0.0125).

T h e slight difference in the regressions (b) of the two dose-response curves

for prednisolone and fluocortolone m a y be d e r i v e d from F i g . 5. T h e suppres­

sive effect o f prednisolone in lower doses, e. g. 20 mg, is more r a p i d in onset

and also more m a r k e d than the effect of fluocortolone, whereas the effect of

fluocortolone appears to last somewhat longer. T h e s e differences in onset of

the suppressive effect d iminish w i t h higher doses. H o w e v e r , the longer d u r a ­

tion of the fluocortolone effect m a y possibly cause a more pronounced sup­

pressive effect of fluocortolone at doses above approximate ly 100 m g (Figs . 3

and 4).

Nei ther prednisolone nor fluocortolone show any fluorescence in the a l c o h o l -

sulphuric a c i d reagent. H o w e v e r , a n important question to be answered is,

whether the metabolites o f these synthetic corticoids interfere w i t h the f l u o r i -

metric determination. T h e fluorescence of serum from adrenalectomized

patients ( F i g . 6) w a s therefore determined for 48 h after the administrat ion

of 50 m g prednisolone, and fluocortolone respectively. N o evidence of any

such fluorescent metabolites was obtained.

p g C O R T I S O L

lOOmg

4 η oo

3 0 C O R T I C O I D

t 6 o o 9 o o 1 2 o o -| goo 2i*°° 6~ 9°° 12°° 18°° 24°° 6°°

Fig. 5. Dynamics of the suppression induced by prednisolone and by fluocortolone.

The mean values ± S D of c ircadian serum Cortisol levels of controls ( N = 25) and of subjects receiving 20 mg prednisolone ( N = 9) or 20 m g fluocortolone ( N = 5) at

7.00 a. m. are shown.

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^ j q C O R T I S O L

1 0 0 m l

^ ^ 700 C O R T I C O I D

ι 0 = 5 0 m g FLUOCORTOLONE

g o o 900 ^2° ° 180 0 2 A ° ° 6 0 0 9°° 12 ° ° I 8 0 0 2 4 0 0 6 ° °

Fig. 6. Basic fluorescence i n tota l ly adrenalectomized patients after 50 mg prednisolone

or fluocortolone at 7.00 a. m. The circadian Cortisol curve of normal controls ( + ) and normal subjects receiving 50 m g prednisolone ( · ) or fluocortolone (o) were compared w i t h the f luor imetr ic values of serum f rom tota l ly adrenalectomized patients ( M . M . , H . M . ) . T o t a l l y adrenal ­ectomized patients were taken off Cortisol replacement therapy 18 h before the f irst serum Cortisol determination (6.00 a. m.) and received 50 m g of prednisolone ( · ) or f luocortolone (o) at 7.00 a. m. No evidence for the presence of any fluorescent meta­bolites of these synthetic corticoids was obtained. The values were close to the basic fluorescence of »cortisolfree« serum (1-2 /fg/100 ml) i n this assay (Kluge et al. 1970).

D I S C U S S I O N

T h e proposed method for ca lculat ing the integral differences of serum C o r t i s o l

levels between c o n t r o l s and subjects receiv ing synthetic corticoids, m a k e i t

possible to quantitate the suppressive effect of corticoids in m a n . I t shows that

any analys i s of the suppressive action of different corticoids s h o u l d take i n t o

account both the degree and d u r a t i o n of s u p p r e s s i o n . T h i s c o n c l u s i o n is

t h o u g h t to b e v a l i d despite t h e r e c e n t observation of Hellman el al. (1970),

that C o r t i s o l is secreted episodically.

T h e approach presented in this paper m a y provide a means of comparing

the suppressive potency of various corticoids. T h e relat ive potencies cannot be

expressed by simple ratios or factors (Fiedner 1967). R a t h e r , dose-response

curves h a v e to be obtained as demonstrated under Results , since the relat ion

between the potencies is not constant throughout the whole range of doses

administered. I t w o u l d appear to be just i f iable to look for analogous dose-

response relationships for other corticoid effects i n m a n , e. g. diabetogenic,

a n t i - i n f l a m m a t o r y , ulcerogenic a n d other actions of corticoids.

It remains , however , an open question whether the suppressive effects of long

515 3 3 *

term corticoid therapy and of single ora l corticoid doses, as described in this

paper, are the same for a l l synthetic corticoids.

A C K N O W L E D G M E N T

The authors are indebted to Mrs . R. Fröhlich for excellent technical assistance.

R E F E R E N C E S

Bethge I L : K l i n . Wschr. 48 (1970) 317. Boss N.y Kluge F., Gerb A. C, Η of mann Η. Sc Scriba P. C: Sympos. Dtsch. Ges.

Endokr. 16 (1970) 368. Gaddum J . I L : Pharmacol. Rev. 5 (1953) 87. Hedner L . P.: J . Endoer. 37 (1967) 57. Hellman L . , Nakada F., Cur Ii ] . , Weitzman E. D., Kr earn ] . , Roffwarg S., Elhnan S.,

Fukushima D. K. Sc Gallagher Τ. F.: J . c l in . Endoer. 30 (1970) 411. Hochheuser W., Gerb Α., Müller-Bar dor ff M. Sc Thiele H.: Dtsch. med. Wschr. 94

(1969) 2488. Kluge F., Gerb A. C , Boss N., Fahlbusch R. Sc Scriba P. C: K l i n . Wschr. 48 (1970)

929. Radvila Α.. Dettwiler W., Rohner R. Sc Studer H.: Schweiz, med. Wschr. 99 (1969) 709. Scriba P. C, Boss N., Gerb A. C, Kluge F. Sc Müller Ο. Α.: 3rd Intern . Congr. H o r ­

mon. Steroids, Excerpta med. (Amst.) I n t e r n . Congr. ser. no. 210 (1970α) 177. Scriba P. C, Gerb A. C, Kluge F., Boss N. Sc Müller Ο. Α.: Ζ. analyt . Chem. 252

(1970/;) 284.

Wallis W. A. Sc Roberts Η. V.: Methoden der Statistik. Rowohlt , Taschenbuch (1969).

Received on November 13th, 1970.

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