Anticoagulant action of glucuronolactone in rat

THROMBOSIS RESEARCH Printed in the

United

311-316, 1972 Press, Inc.

vo1.1, PP* Pergamon

States

~ANTICOAGULAWI ACTION OF GLUCURONOLACTONE IN RAT

V. H. Doctor, B. Shepherd and W. McCullough of Chemistry, Prairie View A&+¶College; Prairie

Department

26.6.1972.

(Received

Accepted

by

Editor

A.L.

View,

TX

Copley)

A single oral dose of 1.5 g/kg of glucuronolactone to rata ABSTRACTS significantly prolonged for 6 days the whole blood clotting times. Since the anticoagulant action is pronounced in ?? iliconized glass tubes, clotting is inhibited at stage(a) after the contact actiAdministration of 1,4-aaccharolactone (a mctabolite of vation. D-glucuronolactone) at 1.5 g/kg also exhibited similar anticoaguAnimals receiving glucuronolactone or 1,4-aaclant properties. 1088 of body weight or charolactnne showed no toxic effects viz. changes in the appearance of the internal organa.

Introduct Die&man

and liopfenapirger

glucuronolactone

and reported

LDso for

rat

(2)

in human beings

that

was

curonolactone

way,

flora. the

is given has other

(8 g/day)

(1) the

Evidence

studied

This

wa8 presented

administration

development alone.

results

to the

on D-glucuronolactone

Studies

also

of D-aaccharic

action

reported

toxicity

the

and Hoffman

(2 g/day)

in intestinal

prevented

of D-

because

by Woolridge

streptomycin

of hypoprothrnmbinemia

The presence

been ascribed tissues

treatment

acute

of streptomycin

in combination

combined

the

compound was innocuous

the eoPergence of post-treatmem

without teric

13 g/kg.

that

ion

and glu-

aterilizatiou

resistance

in the

en-

in some unrecognized

which occurs when streptomycin acid

of a dehydrogenaae,

in normal present

mammalian in the

urine

liver

and

(3,4).

on the value of D-glucuronolactonein rheumetic 311

312

ANTICOAGULANT

and in a variety

of unrelated

diseases

present

report

i.s concerned

with

lactone

in rat

on the clotting

are

breeder

time

lactone

between

(Eastman

1.5 glkg dermic

overnight.

which

animals

solution

treated

were of

morning

30,

60 min)

injected

sodium

obtained

from a local

made into

abdominal

the

drawn from the

lower

poaterior

Pyrex glae8 to

tubes

siliconized tubes.

The

tubes

at half

minute

ed by

inverting

the

in each

group.

were the

were

0.085

with

Kalamazoo,

took effect,

0.2

at 37O

The

was

clotting

were

ml of blood

0.2

in a water times

There given

whole

At 2 hours, blood

clotting

1 day, times

2 days, of the

4 days, rats

were

5 days, recorded

6 days

Per was

was to

ml was transferAdams,

New bath

were the were

York, and

determinlong 6 rats

1.5 g/kg

ronolactone, while the control rats were given the same dose saline.

Mich.1

an incision

experiment,

studies.

rats

aqueous

ml was transferred

rema ining

In a seperate

experimental

of the drugs,

f'Siliclad'f, Clay

incubated

At various

ml of a 10% w/v

0.4

starved

of the drug

by mouth.

Company,

the

at hypo-

group were

suspension

given

Out of this

intervals. tubes.

control

and approximately

cava.

and a blunt

administration

with

(Upjohn

of glucuronolactone The

and the

(13 x 1 cm) while

tilted

effect

rats

following

(treated

a syringe

2 ml of an aqueous

area

Glucurono-

There were 10 rats in each

as the anaesthesia

vena

experiments.

N. Y. 14650) was administered

saline

cyclopal As soon

these

tube.

subcutaneously

100 g body weight.

term

were

by mouth with

in physiological

(15,

for

as a stomach

The following

intervals

N.Y.)

of glucurono-

blood.

150-200g

Rochester,

suapenrion

served

same volume

or the

red

The

and Methods

were used

Kodak Company,

The glucuronolactone

plain

(5,6).

of administration

of whole

between

200-300g

aa an aqueous

needle

group.

the

weighing

impressive

(Check 6 Jones, Houston, Texas 77066) and were fed Purina Rat Chow.

Rats weighing

time

rats

not very

the effect

Materials Albino

Vol.l,No.4

GLUCURONOLACTONE

glucu-

in physiological

and 7 days

the

in plain and siliconized

ANTICOAGULANT

Vol.1 ,No.4

tube6 as described

GLUCURONOLACTONE

above. of 1,4-raccharolactone

An experimant was conducted on the effect (pureha8ed from Sigm8 Chemioal Co., rat on the alottlng toneally There

after

St.

Louis,

tlm8 of whole blood.

at 1.5 g/kg body weight

MO. 63178) administration

The aompoundwas given

The control rats were given

the sam volume as of the drug in physiological

the administration,

and sllfconlted

to

lntraperi-

in the form of 2 ml of an aqueou8 suspension.

were 10 rats in eaah group.

toneally

313

whole blood alottiag

tubes as described

tins

intraperl-

saline.

At 2 hours

vere determined

In plein

above.

Ite8ultr and Dircunslon In Fig.

1 are prere$ed

the reaultr

of the whole blood clotting

timr

I

I

1

IO

I

30

20

MINUTESAFTER

Effect of oral ?? dminl8tration

FIG.

I

I

40

i

50

60

ADMINISTRATION 1

of glucuronolaatone on the clotting tire of whole blood. Ten rat8 in each group were 8tarved overnight. Next day, glwuronolactone was ?? dmlniotered In 2 ml of aqueour 8urperulon by mouth at 1.5 g/kg while the control rat8 were given the 8am8 volua~ of phyrioloThe clotting times were memured in ?? illconlsed tuber. gical 8aline by mouth. in 8iliaonited control

anianls.

clotting

time8

group.

glarr

tube8

At

of glucuronolactone

15, 30 and 60 min after

for the experimental

The point8

fed

animal8

In

the ?? dminirtration

compari8outo whole blood

group was double that of the control

show the mean value

and the vertical

line8 on the graph

ANTICOAGULANT

314

represent

GLUCURONOLACTONE

In Fig.

the stenderd devietion.

I

2 era presented

ruults

the

of a

*

2

I

Vol.l,No.4

3cAY64

5

6

7

FIG. 2 on the clotting time of blood. The sems prosedurs 1 wes uud axcept thet the blood wu drewn et 2 hrs, 1 dey, 2 deys, 4 days, 5 deys, 6 deys end 7 deys

gffeat

wholi

gluauronoleatoae es under Figure

of

of ore1 edministretiou

ud silicouised respsatively eud the alottieg tiues were recorded in plein ?? The results era mean velue of 6 rats with vertiael lfuss representing tha steuderd devietlon. The are8 with the slentlng line8 represent the renge of clotting times obteinsd for the control animsls.

tubse.

timss

long term study where the clotting

of ona mek.

The eree with the slenting

for the control

obteined

?? nimels

showed slgniflaently

lethel

intrevenous

csrdfec

punctures

(1)

delsyed

the clotting

over 8 period

of glucuronolectone

mstsly

The fete

et 1 g/kg in rat

5% of the ccnnpound vu

timss,

pertiaulerly

glucuroin sill-

ore1 dose of glucuronolectoao,

the clotting

tisms

of three hours.

of 8 single

orelly

in the urlno

during

??

teken by

The enimels which is

edministered

has been investlgetod

excreted

timss

from 3 to 5 min for plein

in the present report were given 1.5 g/kg of glucuronolectone

th of the LD50 of 20 g/kg.

period

thet in two of three dogs given

reported

dose of glucuronoleatone,

ran

for a

nimsls receiving The ??

tubes.

?? single

deily

repruent

higher clotting

conised tubes for 6 deys follarfng

Dieahmsnn end Hopfenspirger

lines

which renged

tubes end 4 to 6 rein for slliconised noleatoee

were msesured

(1).

l/13-

dose

Approxi-

the first

24

Vol.l,No.4

ANTICOAGULANT

315

GLUCURONOLACTONE

hour8 while elevated value8 for urine were reported for a period of rix day8 following thin adminirtration. It

her

been portulated that there are two reparate pathway8 by

which blood clotting my

by 8o1mefactor or factor8 which enter the blood from damaged

ir activated ti88ue

be initiated (7). The 800Called extrinric 8y8tel8

while the intrinric

8yrtem ir operative

when contact with rurfacer

(other than the vaecular endothelium)activate8 relative

impOrtam

he8mrtatic

clot,

the clotting oequcnce. The

of the,@ two rehem88 in the formetion or fn the occurrence

of an effective

of thrombo8ir'irrtill not known. The

purpose of rtudying the Whole blood Clotting

time8 in 8iliconir;ed and plain

glur tubea war to determine the nature of the inhibitoryeffect of glucuronolactone adminirtratioa. The re8ultr of Pigr. 1 and 2 indicate that glUCUrOnOlaCtOIIe

inhibit8 Clotting 8ICinlyat 8tCgC(8) after the contact

activation. Marrh(3)

reported that D-glucuronolactoneir converted into 1,4-

?? accharolactoneiruide the an-1

body vta the intermediatefonution of

8CCcharo-(l-4)-(6-3)-dil8ctone.The re8ultr pre8anted in Table 1 rhow that TABLE 1 Effect of Adminirtrationof 1,4-Saccharolactoneon the Whole Blood Clotting Time in Rat

Treatment

Blood Clottirg Time in Min Plain Siliconized

None

4.3 f 6.2

6.5 +, 1.5

1,4-Saccharolactone2 (1.5 g/kg)

5.8 +, 1.3

11.2 +, 1.7

'The figure8 rhow the mean and btandard deviation of 10 rata. 2Tv0 ml of an aqueous su8penrion of 1,4-8accharolactonewe8 given intraperitoneallyand two hr8 later blood was drawn by heart puncture and terted for clotting time in plain and siliconieed tuber.

316

ANTICOAGULANT

1,4-saccharolactone

at 1.5 g/kg significantly

of whole blood in slliconised hyaluronic

acid,

GLUCURONOLACTONE

chondroitin

tubes. sulfate

body which contain glucuronyl.units

prolongs

Vol.l,No.4

the clotting

tims

A number of mucopolysaccharides and heparin are present in its

e.g.

in the animal

backbone however, there is no

evidence that glucuronolactonein the diet can act as precursor for the synthesis

of these compounds. The anticoagulant

reported here may be ascribed

of glucuronolactona

the compound itself

one or more of the proteins or cofactors

soae reaction with the clottfng

to either

effect

entering

into

irrvolved in

sequence or that a mstaboliteviz. 1,4-saccharolactoneformed

from glucuronolactone anticoagulant

action.

inside

the animal body may be responsible

The present study doss not rule out either

for the of these

possibilities. Acknowlednemsnt This work was supported by a grant from the Robert A. Welch Foundation,

Houston, TX. References

1.

DI%HMAN, W. B. and ROPPENSPIRGER,B. The pharmacologic action and acute toxicity of glucuronic acid lactone. Industrial Ued. and Surg.:

20,417. 1951. 2. UOOLIUDCE,W.B. tients

treated

and HDFFMAN,M. Unaltered prothrombfn timss in paAntibiotics and Chemotherapy:l, with oral glucomycln.

249. 1951. 3. MAIM, C. A. Metabolism of D-glucuronolactonein mawsalian systems. Biochem. 5.:99,22. 1966. Hetabollsm of D-glucuronolactone in mawsalian systems. Identification of D-glucarfc acid as a normal constituent of urine. Biochemical 3.:86,77. 1963a.

4. MARSH, C. A.

:'

HODAS, J. H., BRANDON, J., and HAulNEY, J. P. Treatmsnt of rheumatic diseases with glucuronlc acid. J. Lancet:69,385. 1949.

6. SMITH, L. W.

“Glucuronolactone”.

New York, Corn Products Sales Co.

1953.

7. DAVIB, E. W. and RATNOPP,0. D. New York, Academic Press.

1965.

1n:Neurath.

“The Proteins”,

Vol.

3.