Long-term effect of a continuous low dose of megestrol acetate on the genital organs and fertility of female rats

71I

LONC,.-TERM EFFECT OF A CONTINUOUS LOW DOSE OF MEGESTIK)L ACETATE ON THE G ] ~ I T A L ORGANS /tND F E R T I L I T Y OF FEMALE

RATS P.C.Sanwal,

J.K.Pande, a n d Be

Central

s

P.R.Dasgupta,

A m i y a Be K a r

S. S e t t y

Drug Research

Institute,

Lucknow,

Indiq

Receive4 : February 9, 1970 ABSTRACT Daily oral administration (1.5 /~g/anisml) of megestrol acetate (6-mothyl-17~ -acotoxypregna-4.6-dione-3.20 dione) for 12 months did not evoke any noteworthy pondoral or histologic changes in the genital organs of female rats| the pituitary gonadotrophio activity and the estrus cycle continued undisturbede However, certain biochemical alterations were recorded which indicated an impairment of metabolic statue of the uterus. The failure of pregnancy observed in fertility performance tests was m a i n l y d u o t o i n h i b i t i o n of implantation and fetal resorptlon. It is suggested that a depresslon of metabolic status of the uterus by the steroid is crucially involved in implantation failure and fetal resorption.

is

The daily

use

of

nicrodose

currently

considered

synthetic

with

progostational

much i n t e r e s t

as

steroids

a positive

and

I convenient of

fact,

hare

moans s,veral

ostabllshcd

chlormadlnone 3,20

dlono),

for

family

recent the

planning

clinical

and

contraceptive

(6-chloro-17~

acetate mogestrol

acetate

and

birth

contro~.

experimental

efflcaoy

of

In

studios such

point (I-5)

a regime

-acotoxyprogna-4,eodienom

(6-mothyl-17~

-acotoxypregna-n e e

* Communication

no.

1481

with

7I 2

S T ER O I D S

406-diene-3t20-dlone)

t

norgestrel

hydroxy-4-estren-3020-dione)

(174

and

15:5

-~thinyl-13

norethisterone

ethinyl-17~-acetoxy-4-estren-3,20-dione)e continuously in

a

in

micro

substantial

~rovtdo

notable which

proportion

effective

peripheral

point

of

of

impact

rendered

detailed

alone

modus

ooorandi

inimical

do not of

biochemistry

to

clearly

found

sperm

the

micro

dose

may b e it

effects

of

to

genital

organse

Accordingly

e the

present

investigation

long-term

effects

of

genital been of

organs

made

the

and

to

throw

fertility some

of

light

dose

the

the

has

I

a mucus

Nevertheless in

cervical

tubal

and

Fu~the~

physiology

been

and

concerned acetate

An a t t e m p t

of

from

information

megestrol

mode

and

contraceptive

have

ratso

with on

has

contrace~tive

action

MATERIAL . ~ D M E T H O D S

with

bred

regular

ostrus

invostigattono (75~

2°F)

experimental

female

They

under

albino cycle were

uniform

periodo

(4-5 kept

rats

(100-1~0

days) in

husbandry

wo~e

g) o f used

airconditioned conditions

the

in

Institute

this

quarters throughout

the

also

steroids

Colony

time

A I

(3,4)o

on

of

female

on

the

same

cervical

changes

well

method

a micro

the

taken

the

(1,2,4)e

for

important this

be

that

as

at

(1-4)e

regime s luteal

involved is

to

-

ovulation

indicated

passage

the

the

of

been

steroids

0 but

~-

(I7~

inhibit

pregnancy

thus

account

standpoint

long-term

subjects

adequately

a

the

the

revealed

factors

on

of

have

endometrial practical

apparently

is

has

invdetigations

mucus

not

against

action

acetate

These

do

protection

locale

is

doses

/9-ethyl-17

the

e

May 1970

ST ER O ID S

Megestrol the

rate

the

human

acetate

of

1,5

(500

daily

(1)|

acetate

is

chlormadinone

administered

/ug/60

basis

megestrol of

~l/rat

dose

quantitative

was

for

kg)

however,

known

acetate

to

(in

mOnthse

olive

This

chlormadinons the

be

(6)e

orally

12

of

7I 3

The

2,5

acetate

on a

potency

times

controls

at

approximated

progestattonal

about

oil)

lees

received

of

than the

that

vehicle

alonee A group feeding|

of

their

weighed

to

tissues

were

were the

the

uterus

of

previously procedure

of

and

in

soluble

fertility

performance

according

pregnant note.

the

then

animals

allowed

to

of go

WtlderOs were

removed

eosine

and

The

paraffin

sections

Sections

ret~culin

of

staininge

the

same

was

ddtermined

as

employed by

triphosphatase

commenced

steroid

(or

procedure

olive

of

laparotomized

implantations to

and

steroid

balances

serial

adenosine

test

the

were

wore

Roller-Smith and

last

the activity

e__tt a~l ( 9 ) e

month to

number

and

Umbreit

12

to

the

pituitary

phosphorus

(8)

The

the

a

after

hematoxylin

of

of

the

analyses

e~t a l

hree

fluid

subjected

Acid

Hawk

and

Bouin's

~hrlich's

also

24

mg i n

technique

conducted

to

0o2

the

completion

The

nearest

biochemical

(7)o

killed

organs

with were

was

genital

fixed

stained

Methods

by

animals

term,

The

days

oil)

Kar and on

and

15

day

to

couree~ Betty

10

condition

steroid

prior

course

the

and

was

(10)o

of

p~egnancy

of

the

fetuses

continued

till

parturitions

by

The

total

gonadotrophin

the

immature

mouse

content

uterine

weight

of

the

pituitary

method

(ll)e

was

assayed

0

7 1~

S T 1~ R O I D S

i.

15:5

i=11 --~ -o =I hi 0

~

he u

all

WO

O~

nl m ad go ,l~ f,,o

•

ePl I

~/

ellD 1"41

~

I

~

0

I

o

()

M~

°~

P I

1 !

"gig Z h

k

Q

o

•

I t

May 1970

ST ER O I D S

715

RESULTS It

will

be

mogeetrol

seen

from

acetate

had

and

latter

duration

The sad

the

the p i t u i t a r y !

kistelogie

stermid

rats.

in

the

in

generals

was also

in

uegestrel

aeatate

in

the

wells

appeared

8troaa.

The

endouetriuu Of

of

signtfieantly

reduoed

acid

skewed

and

significantly lactic The that

and

results

8g~ of

eorrespomdins 505.

lactic (vs.

aeid

The

of

(~se

number

of

of

and

adenosine

trend.

On t h e

dekydroKonase

control for

P•

other

uterus

those tramstn

the

by the

steroid.

2)e

activity 0.01

the were

respective-

aettvity

hand e glucose-O-

There

inereased was

no effect

on

eoneemtratiene teat

positive testate

sites

frequent

the

(Table

and

activities

shoved

However.

seen

of

disturbed

uterus,

more

oo1.18 w a s

0.05

lutea

decid~al

tripkesphatase

O•O1)•

megostrol

of

(Table

3) s h o w e d

~tiaKm

whereas

treated

o oorpmra

ones

lutea

the

control

Some o f

pieephatase

perfollanee

implant,ties

yore

the

phosphorus

rate

figures

framework not

eentrols,

soluble

eontrolse

of

unaltered

the

the

decidual

were

P•

of

stage.

the

in

histology

prowess

reticulin

were

aorpera

of

and,alkaline

fertility

figaro

the

of

eontrol4m

acid

the

In

of

normal

umtestru8

eonstituents

pkosphatase

that

mitotic

the and

of The

to

animals

Klyeogan

a diminished

phosphate

similar

vaseularity

concentration

ly)|

were

presence

that

content

ovary

of. the

bioehemlaal

gomadotraphin

of

the

I

on weight

days)

aeeuuulatiom

its

Table

(4-5

during

integrity and

the

fed

in

cycle

noteworthy.

epltheloid

Snail

offset the

oomparable

sells

presented

estrus

The

Ermup was

stromal

formation.

of

features

treated

latter

results

no significant

genital o r g a n s and

the

was

and

the only

the

•

718

ST E R O I D S

15:5

ZlA ¢0

m

0~.*

..q al q~

al k

Ih

Q

al s:

| o. ~1 k O~ ,.cOil*.* 4.~ I I 0 ~,.*

@

,,-I

0 q.4 ,1~

ICl ."~

0 o t~ aS o q,4

®

Cq

*~

0

,.4

! *k ~ 0 ~ ~ uO ~,4~ ~0

m

4,) m.4

0 0

M IS *~ e

0

O~

O O

N

M

01 481

aO 0 ,.4

II ill

nl et fl 0' .D el

o

0

e,l Ol

u

q.4w4 ,,-4 0 I~ 0 eO Jl~ Jl • k 4 ~ ml 00~S8 Z ~ e

t,,4'

ell ~] q.II

o.l'

t 0

0

aS,~ pl

II ,..4 II, n

M I1 ~I ~

k •I

*q.I O~ m 0 0 J~ O O.e4 ~Z: N m ~ k~

,

II ml

0

O! Ill

k ~-

,4~! OI [,..41

el ,V 0 ~.c

I eel 0 o ,~ I I iIS .,4 4~ O~

el

0 m .~ • 4~4~ ~ ~s

M

m

.| 0 ~

Zl~ O k m II.O il.

w,lv 0 •

II 4~ II 4~

0 ~ k • MQ O0 r~eo

~

t. o

%

iii

0

0

May 1970

(from

ST ER O I D S

NAD) w h i c h

oxidation

o£

respiratory

in

a

tissue

lo88

minimum. mien

It

of

NADI~

eonverslon

for

of

operative change

seems

in

in

acid

be

intriguing.

components

involved

exhaustion

due

dose

regime

This

may very the

dose

to

that the

pituitary

glycogen

the

known

to

Is

production

to

keep

its

steroid)

pathways

down

of

tea.

utillzao

processespincludlng

and

in

well

it. the

continuous

be

the

finally

is

to

g!ycogen

unaccompanied

-- o v a r i a n

as is

at

the

observed for

fertility interplay

and

are

light

of

by

this. in

this

this

stimulation

e

ere

any

of

of

o£

the

performance £s

generally

seems, to

enzyme

undergo

steroid.

contlnuous

in

It micro

uterus

is

morphologlc

consistently

contraception

study

decrease

cellular

the

acid

effects

marked

the

the

of

the

vlth

status

basis

the

that

o£

influence

stimulation documented

conceivable

synthesis

so

a

well

observed

metabolic

daily of

uterus In

activity

overall

prosestosens results

of

is

energy

the

enter

HAD t h e r e b y

cycle

uterus of

phosphatase

of

under

endometrtum

The

the

of

alternatlv~

acid

alkaline

However.

appears

pathway

nolle

to

Krebs

for

cannot

production the

biosynthetic

(15,16,17).

phosphatase

thus

hydrogen

where

influence that

to

activity

progoetogens

of

unllkely

NADI~[ w h i c h

energy

of

chain

level. is

of

respiratory

its

of

part

persistent

depletion

A reduction phosphatase

the

reductive

protein

words.

circuitous

on

pyruvic

since

other

scheme

a

attempt to

the

conditions

Such

(due

enters

transfers

This

under

(14).

signlfy~an

energy

In chain

NADH ( 1 3 ) o

unoperatlve may

substrates. oxidative

producing occur

eventually

719

deprossede

'suppression'

women

taking

low

(l.2)e tests not

show

that

the

disturbed

by

cyclic

7z6

ST E R O I D S

o

~, •

15:5

~^,-,

=J= =n ~, . 0 oet .: :0. :? .~ ®. =~ e..,. lO

:=

-o

,~.

o:

.~

qlp4 Ill

t~eq •

I~ I

,,.

•

4t,,,4

~

^

01 I~ I

= ="

s-

o

h~

I~

o

~lm

=.

:

=

=,.

~,;

~

~

~

~

~

"

May 1970

ST ER O I D S

offspring on

day

born I0

of

were

substantially

pregnancy

redueede

revealed

steroid

group

were

in

the

corpora

lutea

appeared

to

that

process be

7I 7

Laparotomlc

many

of

of

the

examlnatlon

fetuses

resorption!

in

however,

morphologically

the

their

normale

DISCUSSION The

results

of

the

of

a micro

administration does in

not the

provoke

genital

formation activity Similar this

organs

the

estrus

observations

concentration

hand I of

of

estrogens the

are

uterus

that

or

for

The

pituitary

energy

generally in

elinica]

does

Thus

a decrease

levelt

the

dehydrogenases

coupling

phosphate

which

can

metabolism enter

utlltsed

lactate|

by during

lactic

acid

some

It

for

is of

in nay

with

pertinent this

enzyme.

activity

denote

pathways production

of

a

of of

NADH.

the oxidative respiratory chain for elaboration of

ATPe N A D I ~ w h i c h be

via

increase

glycolytic

influence

together

activity

and

carbohydrate

with

Inglycogen

mechanisms

glucoso-6-phosphate

and

indeed

suggest

a marked

pontose

trans-

studies

activity

Similarly.

of

changes

undisturbede

regime

stimulate

lactic

mouths

~l-4)e

production to

12

decidual

dose

acid

for

gonadotrophic

triphosphatase lactic

oral

hlstologic

focal

recorded

micro

daily acetate

except

uterus.

of

known

(12)e

megestrol

steroids

adenosine

the

of

remaim

been

the

the

constancy

interruption that

have

and

virtual

cycle

progestattonal

other

show

ponderal

rats

and

biochemistry

the

of

endometrium.

other

study

noteworthy

the

O~ t h e

in

any

dose

of

and

the

present

is g e n e r a t e d

lactic

in the

dehydrogenase

oxidation

of

lactate

pentose for to

phosphate

conversion pyruvate

of

cycle pyruvate

NAD8 i s

can to

produced

720

S T E R O I D S

ne|ostrol

acetate.

This

experinentaX

reports

oontributory

to

study: (c)

(8)

fetal

boon

the

aduinistr&tion observed

in

of

(5)0

Inhibition

of

particularly of

implant o a substantial

proportion

in

mating'by after

has

show that o2 the

to

be

present

failure

or

the

soon

the

this

oral

of

and

tatters

Implantation

implantation

data

olinlcel

observed

irrespective

present

with

obvious

failure (b)

Failure

The

Three

mating

spooles

(5).

sodloutish

rats

this

eontorm~ty

Pr0gnancy

resorption. in

in

(ll2e4ei)e

inhibition

recorded

is

15:5

&nd

steroid

has

paronteral also

route

of

• yon if fetuses

been

ova somehow

undorKo

resorption. The deranKenont oructally of

involved

maEestrol

a most

unsuitable

this

be

reduced

experiment

subjected It

implantation

the

of

ovulation

that and

steroid

trotted or

luteal

proKestorono

of

Kroup.

This

doKenorotion

lnvestiKntion

oklormadinone

is

also

known that

to and

corpora

to

dose

sustenanoo

(18)e

shauKos

micro

uteri

may c o n t r i b u t e

any ultrastructural daily

effects

suoh

Insufficiency.

oltnlonl

fetal

has

resorption number

may b e

pro,sis

whisk

fetal

uterus

and

ova transport

total

a rocsnt

the

consideration

(8),

and

of

ooaceivable

due

the

the

endogenous

record

is

for

in

situation,

(10).

that

failure

inhibition

to

It

status

antilnplantatloa

nay bedisturbed

in

the

the

Nevertkelesse

implantation

,viral1

in

bed

possibility

function

moUth,lie

acetates

prepancy. the

of

in

some i n h i b i t i o n

the

will of

be Elven oorpora to

In

point

has

boon

loadtnK

neonate

of

lutes

of

fast,

found

to

Whatever k&o b e e n

to

the

may s i f n l f y

corpora

be

either an may b e

uablo women

treatment

o~ o v u l a t i o n

to

does

May 1970

ST ER O I D S

Indeed o c c u r In suoh p a t i e n t s

72 !

(1)o

ACKMOWLEDG~iEMTS This

investigation

of Health0

was s u p p o r t e d

Family Planning

and t h e F o r d F o u n d a t i o n , Dhar f o r h i s

Interest

by g r a n t s

and Urban D e v e l o p m e n t e G o v t . o f l n d t a The a u t h o r s

in this

study.

are

of M e s s r s . B r i t i s h

Kilt

grateful

t o D r , MoLe

One o f t h e a u t h o r s

t8 a S e n i o r R e s e a r c h F e l l o w o f t h e I n d i a n R e s e a r c h . The g e n e r o u s

from t h e M i n i s t r y

(PoC.S.)

Council of Agricultural

of meKegtrol acetate

by Dr. DoTeModi

Drug H o u s e , Bombay i s a c k n o w l e d g e d . REFERI~CES

lo

Martinez-SJganautou0 J o e G t n e r - V o l a s q u e s e J e s C o r t a s ' G a l l e g ° s e Yes Asman0 Rot R a j a s , B e , G u l h e r e . s - N a J a r , A. and R u d e l , He, BRIT. MEDo Jo 4 , 730 ( 1 9 6 7 ) o

20

Z a n a r t u , J o t R o d r i g u e z - M o o r e , Go, P u p k i n , .Me, S a l a s e O, and G u o r r e r o . Roe BRIT. MED, J , ~e ZSS ( 1 9 6 8 ) o

3.

Foes,

40

M e a t s , E.~ V e s s e y e MOP.. A n d o l s e k , L. and Oven, A . , BRIT. MED. J . 4 , 730 ( 1 9 6 9 ) .

5.

Chang,

6.

G r e o n b l a t t , R . B . and Mahesh, 140 321 ( 1 9 6 5 ) .

7.

Panda,

G . L . , S v e n d s e n t EoK. e F o t h e r b y , BRITo MEDo J . ~o 489 ( 1 9 6 8 ) .

C.C. and K l n c l ,

J.K.,

Snocl.

F.A.,

Ko and R t c h a r d s e D . J o :

STEROIDS. 1 2 , 189 ( 1 9 6 8 ) . V . B . , METABOLISM.

D a s g u p t a . P . R . and K a r , A . B . , 2 . 156 ( 1 9 8 9 ) .

GEN. COMP. ]DIDOCR.

8.

Hawk, P . B . , O s e r , B . L . and Summers,n, W.H., PRACTICAL PHYSIOLOGICAL CHEMISTRY, ( 1 9 5 4 ) , H c G r o w - H i l l C o . , New Y o r k .

9.

U a b r o l t , W.W.I B u r r i g , R . H . a n d S t a u f f e r , J . F . , MANOMETRIC TECHNIQUES. ( 1 9 5 7 ) , B u r g e s s P V b l . C o . , M l n n e a p o l l s o

I0.

Kar,

A.B. and S o r r y ,

B.S.,

INDIAN J .

MED. RES. 53,

1080 ( 1 9 6 5 ) .

II.

Kar,

A . B . , Chowdhury e S . R . , Chowdhury, A . R . , Kambo3t V . P . and C h a n d r a , H. I STEROIDS. ~ . 519 ( 1 9 6 5 ) .

722

ST E R O I D S

IRe G r o s s I M. I F ~ F L ,

15:5

STERL, l Z s 245 ( l g 6 1 ) 0

1 3 , H e l l n a n . S o . I n NON GLYCOLYTIC PATHWAYS OF METABOLISM OF GLUCOSE. A c a d e u t c P r e s s I n c . ( 1 9 6 4 ) , Now Y o r k . 14. Wllsonl

E,W,. J.

ENDOCR, 44 e 63 ( 1 S 0 9 ) 0

~5e G e l d b e r g 0 Be a : d J O l O l J r o j 7~ S4a ( 1 9 S e ) ,

HoWo~ O B J .

GYNKCe

160 G a r c i a - B u n u o l e Re a n d B r a n d e s e Do 0 AM, J . 94.

~STe

GYNEC,

1045 ( 1 9 4 6 ) 0

1 7 . HuEhes t EoC. 0 J a c o b s p R . D . and R u b a Z l s I A. s AM. J . GYNEC. 8 9 . 50 ( 1 9 6 4 ) . 18o P / a e u s ,

OBST.

Go, TI-IE COHTROL OF FERTILITY. A c a d e m i c P r e s s

Inc.i

( 1 9 6 S ) t Hew Y o r k .

1 9 . M a r q m e s - M o n t e r e H. I G u t t l o r r o s - N a J a r t A. t A g n e r . Roe ~ n e r - Y e l a z q u e s e J o j R u d o l e H. and M a r t i n e s - M a n a u t o u e J o e FERTL. STERL. I_.99, 3e3 ( l e e S ) .