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EXPERIME NTA L MO DELS AN D PLATELET REG ULAT OR Y DR UGS 223

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THEEFFECTIVENESS OF DRUGS THAT

MODIFY PLATELET FUNCTION IN

PERIPHERAL VENOUSTHROMBOSIS

AND PULMONARY EMBOLISM

A. G. G. TURPIE

McMaster University,

Hamilton , onta rio, Canada

Introduction

Venous thrombi consist mainly of fibrin and trapped red blood cells but there is

evidenc e that some venou s thrombi originate in valve pockets as aggregates of

platelets which propagate with the formation of fibrin to form classical red thrombi

(Paterson , 1969; Sevitt , 1973). In addition, platelet thrombi may form at sites ofdirect

injury to veins, for exarnple, after trauma to the legs or to the femoral vein during hip

surgery (Clagett , Brier , RosofT, Schneider & Salzman, 1974; Clagett , Scheider, RosofT

& Salzman, 1975). It is possible , becau se of the role of platelets in the genesis of

venous thrombosis, that drugs that suppress platelet function may be of value in

prevent ing the formation of venous thrombi which arise principall y as platelet

aggregates. Four ant iplatelet drugs, aspirin, dipyridamole, hydro xychloroquine and

sulph inp yrazone have been tested in prospective clin ical tr ials for the prevention of

deep vein th rombosis, and in one study the efTect of aspirin on fatal pulmonary

embolism was evaluated. Dextran, a glucose polymer introduced as a volume

expander, also inhibits platelet function and has antithrombotic properties, but these

properties may not be due to its antiplatelet efTect alone.

In evaluating stud ies of venou s thrombosis prophylaxis, it is important to consider

the endpoints used to detect venous thrombosis and since clinical diagnosis is

notoriously inaccurate, result s of trials using clinical endpoints are not acceptable.

Three objective and accurate methods of diagnosing venous thrombosis are

acceptable for use in c1inical trials. These are i) the use of leg scann ing in patients

who have general surgical procedures or in medical pat ients ii) the combined use of

leg scanning and impedance plethysmography in patients who undergo hip surgery

iii) the use of venography in all patients (Hull & Hirsh, 1979).

In addition , onl y the result s of clinical trials that fulfill well-defined design criteria

(Genton, Gent, Hirsh & Harker, 1975) can be used to draw valid conclusions and will

be cons idered in this review.

The characteristics of the antiplatelet drugs evaluated in c1inical trials have been

investigated in vitro and in animaI model s of thrombosis. They have also been

studied in man by measuring their efTects on the bleeding time and on platelet

VENOUS THROMBOEMBOLISM AND ANTIPLATELET DRUGS 225

survival time and turnover. Each drug has different relative effects when measured by

these tests . Some are effective in vitro while others show greater effect in vivo and at

present, there is no test that can predict which antiplatelet drugs will be cIinically

effective in man. In addition, although the drugs that have been evaluated cIinically

inhibit one or more platelet reactions and are antithrombotic in animal models, each

has to be tested in prospective cIinical trials to determine its usefulness as a

therapeutic agent.

Mode of action of antiplatelet drugs

Aspirin

The mechanism by which aspirin inhibits platelet function has recently been defmed.

Aspirin inhibits the synthesis of cyclic endoperoxides and thromboxane Al from

platelet membrane arachidonic acid (Smith & Willis, 1971; Roth & Majerus, 1975).

Thromboxane Al is a potent stimulus ofthe platelet release reaction and aggregation

and a powerful vasoconstrictor. Roth & Majerus (1975) demonstrated that the effect

of aspirin was primarily due to its acetylation of the platelet enzyme cyclooxygenase. Rosenberg, Girnber-Phillips, Groblewski, Davison, Phillips, Goralnick &

Cahill (1971) dernonstrated that aspirin acetylates other platelet membrane proteins

and inhibits the platelet membrane enzyme, collagen-glucosal transferase. These

effects will also contribute to aspirin's inhibitory effect on platelet function . Although

aspirin is rapidly cIeared from the circulation, it has an irreversible effect on all ofthe

platelets that are circulating at the time ofits administration because platelets cannot

regenerate cyclo-oxygenase that has been acetylated by aspirin.

Aspirin also inhibits prostagiandin synthesis by blood vessel walls . This could have

important practical implications because prostacycIin, (PGI2) which is a potent

inhibitor ofplatelet aggregation and a vasodilator, is synthesized by endothelial cells

in response to a number of stimuli which incIude thrombin and bradykinin

(Moncada, Higgs & Vane, 1977). It is therefore possible that aspirin could inhibit an

important protective mechanism, although it has been recently demonstrated by

Baenziger, Dillender & Majerus (1977) that much higher doses ofaspirin are required

to inhibit prostagiandin synthesis by cultured vessel wall cells than by platelets.

These in vitro fmdings are supported by the results of in viva experiments in animals

by Kelton, Hirsh, Carter & Buchanan (1978) who demonstrated that a dose ofaspirin

that augmented thrombosis was much greater than the dose required to inhibit

thrombus formation .

Asp irin has been extensively tested for its antithrombotic effects in models of

experimental venous thrombosis in animals. Renaud & Godu (1970) produced

thrombosis in rats by infusing endotoxin and demonstrated that aspirinized animals

had a lower incidence of hepatic vein thrombosis than controls. Petersen & Zucker

(1970) reported that aspirin significantly reduced the incidence of thrombosis in

chemically-damaged rabbit femoral veins . In contrast, however, Arfors, Bergqvist &

Tangen (1975) failed to demonstrate any effect with aspirin using a similar experimental thrombosis model. Kelton et a/. (1978) examined the effect of aspirin on the

size ofvenous thrombi produced by mechanical damage to jugular veins in rabbits. In

this study, thrombus size was quantitated by measuring IlsI-labelled fibrinogen

incorporation into the thrombi and aspirin reduced the size of the thrombi but the

effect was limited to male animals. These observations are of particular interest since

they are consistent with recent cIinical reports which demonstrated that the effectiveness of aspirin in preventing thromboembolism in patients with hip replacement

(Harris, Salzman, Athanasoulis, Waltman & De Sanctis, 1977) and with transient

cerebral ischemia (The Canadian Cooperative Study Group, 1978) was limited to

males.

226 A. G. G. TVRPIE

Aspirin in doses of300 mg prolongs the bleeding time for up to five days in normal

volunteers (Mielke , Ramos & Britten, 1973) but unlike other drugs which suppress

platelet function, aspirin does not prolong reduced platelet survival seen in a number

ofthromboembolic diseases (Harker & Slichter, 1972).

Aspirin has been evaluated as an antithrombotic drug in venous thromboembolism, in humans, in doses of300 to 3500 mg daily .

Dipyridamole

Dipyridamole is a pyridimo-pyrimidine compound that was introduced c1inically as

a vasodilator. Dipyridamole inhibits adenosine diphosphate-induced platelet

aggregation (Emmons, Harrison, Honour & MitcheII, 1965) and in high concentrations inhibits the release reaction induced by collagen , adrenaline and thrombin

(Zucker & Peterson, 1970; Cucuianu, Nishizawa & Mustard, 1971).

Mills & Smith (1971) demonstrated that dipyridamole inh ibits phosphodiesterase

activity in platelets and so increases platelet cyclic AMP levels which in turn are

associated with inhibition ofplatelet function. It has been reported that dipyridamole

has a greater ex vivo than in vitro etTect in platelet function which has been related to

the ant icoagulants used in the in vitro measurement ofplatelet function (Buchanan &

Hirsh, 1978) and to the ditTerences in the albumin concentrations when platelets are

tested ex vivo compared with in vitro (Jergensen & StotTersen, 1978). Dipyridamole

has also been reported to have other etTects on platelet function . Ally, Manku,

Horrobin, Morgan, Karmazin & Karmali (1977) demonstrated that dipyridamole

inhibits thromboxane A2 synthesis, although Moncada & Korbut (1978) presented

evidence contrary to this, and Best, McGuier, Jones, Holland, Martin, Preston, Segal

& Russell (1979)demonstrated that dipyridamole inhibits prostagiandin synthesis.

Dipyridamole has mainly been tested in animal models of arterial thrombosis and

has proven only moderately etTective (Arfors et al., 1975).

Dipyridamole in doses of400 mg daily prolongs shortened platelet survival seen in

patients with a number of thrombotic diseases and when combined with aspirin

100 mg daily of dipyridamole prolongs platelet survival in these patients, wherea s

when given in doses of200 mg alone it has no etTect (Harker & Slichter, 1972; Harker,

Slichter, Scott & Ross, 1974).

Dipyridamole has been evaluated for its antithrombotic etTect in man, singly and

in combination with aspirin using the doses that have been demonstrated to prolong

platelet survival.

Hydroxychloroquine

Hydroxychloroquine is an antimalarial drug that inhibits platelet function . Madow

(1960)demonstrated that red cell sludging was reduced by several antimalarial agents

particular1y hydroxychloroquine and subsequently, Carter, Eban & Perrett (1971)

demonstrated inhibition of ADP-induced platelet aggregation by hydroxychloroquine in vitro although Pileher (1975)did not demonstrate any ex vivo change in ADP

or adrenaline-induced platelet aggregation after hydroxychloroquine ingestion. The

mechanism ofthe inhibitory etTect ofhydroxychloroquine and platelet function is not

known . Hydroxychloroquine has no etTect on the bleeding time and does not prolong

platelet survival.

Hydroxychloroquine has been evaluated as an antithrombotic agent for venous

thrombosis prophylaxis in doses of600 to 1000 mg daily .

Sulphinpyrazone

Sulphinpyrazone is a non-steroidal anti-inflamrnatory drug which was introduced as

a uricosuric agent. Sulphinpyrazone inhibits platelet aggregation by adrenaline and

VENO US THROMBOEMBOLlSM AND A NTIPLATELET DRUGS 227

collagen (Wylie, Chesterman, Morgan & Ca staldi, 1979) but there are conflicting

reports on its effect on ADP-induced platelet aggregation. Sulphinpyrazone also

inhibits platelet adhesion to coll agen in flowing blood but, at ph ysiological

haematocrit level s, inhibition of adhesion requires concentrations o f sulphinpyrazone that are unl ikel y to be achieved in humans (Davies, Essien , Cazenave,

Kinlough-Rathbone, Gent & Mu stard, 1979). The mode ofact ion ofsulphinpyrazone

is incompletely understood but there is evidence that sulphinpy ra zone is a

competitive inhibitor of cyclo-oxygenase (Ali & Ma cDonald, 1977). There is

evidence that a metabolite of sulphinpyrazone may be a more potent inh ibitor of

platelet aggregat ion than the parent compound. Buchanan, Ro senfeld & Hirsch

(1978) reported that platelet aggregation in vivo was inhibited for up to 18 h following

administration ofsulphinpyra zon e at whi ch time the drug had been cleared from the

plasma for 14 h. In addition, when normal platel ets were incubated with plasma from

rabbits pretreated with sulphinpyrazone 18 h previou sly, a signi ficant inhibition of

aggregation was found .

Arfors et al. (1975) demonstrated that sulphinpy razone did not prevent venous

th rombosis produced in rabbit femoral vein s by a combination ofendothelial damage

a nd stasis. Sulphinpyrazone prolongs reduced platelet survival in pati ents with gout

(Sm ythe, Ogruyzlo, Murphy & Mu stard, 1965) and a variet y of diseases associated

with thrombosis (We ily & Genton, 1970).

Sulphinpyrazone has been eva luated in two clinical tria ls for venous th rombosis

prophylax is in dose s of800 mg daily.

CIinical evaluation ofplatelet suppressant drugs in venous thromboembolism

Aspirin

Aspirin has been evaluated for the prevention of venous thromboembolism, either

alone or in combination with other antiplatelet drugs, in patients undergoing genera l

abdominothorac ic surger y or orthopaedic procedures. In general, aspirin has not

been effective in reducing the frequency of veno us thrombosis in general surgica l

patients and the results have been mixed in th e patients undergo ing orthopaedic

procedures. The results are summa rized in Table I.

Table 1 Random ized studies using aspirin in the prevention ofvenous thrombosis.

N umber Venous thrombosis(%)

0/

Author Dose tmg) Su rgery patients Control Treated

MedicalResearch Council (1972) 600 General 303 22 28

Clagett et al. (1 974) 1300 General 105 20 12

Shöndorf& Hey (19 77) 450 Orthopaedic 45 60 53

Har ris et al. (1 977) 1200 Orth opaedic 95 43 25

Hume et al. (19 77) 1200 Orth opaedic 71 35 30

McKenna et al.(1980) 900 Orthopaedic 43 75 78

3500 (knee) 8

Two studies on th e effect of aspirin, 600 mg and 2400 mg (ü'Brien , Tulevsk i &

Etherington, 1971) and 600 mg (Medical Research Council, 1972), for th e pr evention

of leg scan-detected venous thrombosis in patients undergoing thoracotomy a nd

general surgery showed no benefit from asp ir in. Similarl y, Clagett et al. (1974) found

no benefit with 1300 mg ofaspirin daily in gen eral surgica l patients using leg scanning

to detect venous thrombosis with venogra phic confirrnation. On the first an al ysis of

the dat a, th ere was 00 significa nt difference between the incidenc e of venous

228 A. G. G. TURPIE

thrombosis in the control group (20%) compared with the treated group (13%) but,

after 4 patients in the aspirin group in whom venous thrombosis developed were

excluded because they did not receive the drug, the difference became statistically

significant.

A combination ofaspirin and dipyridamole has been reported to produce a signifi -

cant reduction in the incidence of postoperative venous thrombosis detected by leg

scanning in general surgical patients by Renney, O'Sullivan & Burke (1976).

There have been a number of trials on the use of aspirin in preventing venous

thrombosis inpatients undergoing surgery for hip fracture or following elective hip

replacement. Salzman, Harris & Oe Sanctis (1971) reported on a comparison of

aspirin, dipyridamole, warfarin and low-rnolecular weight dextran, using clinical

diagnosis as the endpoint, and reported that aspirin was as effective as warfarin, but

that dipyridamole was ineffective. In a second comparison using venography to

detect venous thrombosis (Harris et al., 1974) heparin was less effective than the other

three treatments but the incidence of thrombosis was high in all groups and in the

aspirin-treated group, there were more large and multiple thrombi. Harris et al.

(1977) reported the results of a prospective control double-blind study of aspirin,

1200 mg daily, in patients undergoing total hip replacement using venography as the

diagnostic endpoint. Venous thromboembolism developed in 11 of 44 patients

receiving aspirin compared with 23 of 51 patients receiving placebo. This difference

is significantly different. In this study, the beneficial effect of aspirin was limited to

men; 4 of23 men who received aspirin and 14 of23 who received placebo developed

venous thrombosis compared with 7 of 21 women who received aspirin and 9 or 26

women who received placebo. Review ofthe previous study by these authors showed

a similar differential benefit between men and women. Hume, Bierbaum, Kuriakose

& Surprenant (1977) reported the results of a non-randornized trial of aspirin

(1200 mg daily) in elective hip surgery, in which leg scanning and impedance plethysmography and venography were used to diagnose venous thrombosis and the

frequency was significantly lower in the aspirin-treated group compared with the

contro!. Sch öndorf & Hey (1978) reported that intravenous aspirin in doses of

900 mg had no effect on leg scan and venographically detected venous thrombosis in

patients undergoing elective hip surgery.

A study reported by McKenna, Galante, Bachman, Wallace, Kaushai & Meredith

(1980) in patients undergoing elective knee surgery is of particular interest as aspirin

was ineffective when used in a dose of 900 mg daily, but markedly reduced the

frequency of objectively detected venous thrombosis when used in a dose of 3500 mg

daily. The reason for this is not certain.

Zekert, Kohn, Vormittag, Poigenfurst & Thien (1974) reported the results of a

randomized double-blind trial of aspirin (1500 mg da ily) for the prevention of postoperative pulmonary embolism following surgery for hip fracture. In this study,

pulmonary embolism was one of the main endpoints, and all patients who died in

hospital had an autopsy. One hundred and twenty patients completed the study and

12 patients died, 3 in the aspirin-treated group and 9 in the placebo group. Although

there was no statistical difference in the total number ofdeaths, pulmonary embolism

was found in I patient treated with aspirin and in 8 patients treated with placebo. In

each case, pulmonary embolism was considered to be the cause of death. There was,

in addition, a reduced incidence of clinically diagnosed deep vein thrombosis and

pulmonary embolism in the aspirin-treated group. Thirty eight patients were

excluded from analysis of the study for a variety of reasons and 8 of them died; a

mortality four times greater than in the patients included in the study. Jennings,

Harris & Sarmiento (1976) reported on the use of 1200 mg of aspirin daily in patients

undergoing total hip replacement and found no pulmonary emboli in patients in

whom the expected incidence ofdeath from pulmonary embolism was 1-2%.

Oechavanne, Ville, Viala, Kher, Faivre, Pousset & Dejour (1975) and Silvergleid,

VENOUS THROMBOEMBOLlSM A ND ANTIPLATELET DRUGS 229

Bernstein, Burton, Tanner, Silverman & Schrier (1977) found that the combination of

aspirin and dipyridamole did not reduce the incidence of venous thrombosis after

elective hip replacement. Morris & Mitchell (1977) reported that daily doses of

900 mg aspirin combined with 300 mg dipyridamole, or of flurbiprofen 150 mg

or dipyridamole of 300 mg did not reduce the frequency of isotopically diagnosed

venous thrombosis in patients with hip fractures, when compared with control

patients receiving no treatment, but in this study, since leg scanning was used to

detect venous thrombosis, no account was made for the possible occurrence of

isolated thrombi at the fracture sites.

Dipyridamole

An early study wh ich evaluated the effect of dipyridamole on postoperative venous

thrombosis used a c1inical endpoint to detect venous thrombi and failed to demonstrate any benefit (Browse & Hall, 1969). A number ofstudies have been carried out

comparing dipyridamole alone or in combination with aspirin (see above). The study

by Morris & Mitchell (1977) compared the efficacy of low-dose heparin, dipyridamole alone, and dipyridamole in combination with aspirin and flurbiprofen in

preventing venous thrombosis using leg scanning in hip fracture patients, none of

these regimens significantly reduced the incidence of venous thrombosis.

Dechavanne et al. (1975) reported that low-dose heparin was more effective than

dipyridamole plus aspirin in leg scan-detected venous thrombosis following hip

replacement. In contrast, both Renney et al. (1976) and Plante, Boneu, Vaysse,

Barret, Gouzi & Bierme (1979) reported that aspirin plus dipyridamole was effective

in decreasing leg scan-detected venous thrombosis following elective general surgery.

Hydroxychloroquine

There have been a number of c1inical trials on the use of hydroxychloroquine for the

prevention of venous thrombosis in general surgical and orthopaedic patients. The

results are summarized in Tab!e 2. In each of the three trials involving general

surgical patients (Carter et al., 1971; Carter & Eban, 1974; Wu, Tsapogas & Jordan,

1977) where the risk of thrombosis would be considered moderate, a significant

reduction in deep vein thrombosis was detected in the hydroxychloroquine-treated

patients. Conflicting results have been reported in the studies of patients undergoing

orthopaedic procedures. Chrisman, Snook, Wilson & Short (1976) in a comparison of

hydroxychloroquine and placebo for the prevention of venous thrornbosis, in

patients with fractures or orthopaedic procedures to the knee or pelvis, reported that

venous thrombosis, diagnosed by impedance plethysmography and confirrned by

venography, was significantly reduced in the hydroxychloroquine group compared

Table 2 Randomized studies using hydroxychloroquine in the prevention of venous

thrombosis.

Venous thrombosis (%)

Author

Carter et al. (1971)

Carter & Eban (1974)

Wu et al. (1977)

Chrisman et al. (1976)

Hansen et al. (1976)

Hume et al. (1977)

Cooke et al. (1977)

Dose (mg)

600

800

600

600

600

600

1000

Surgery

General

General

General

Orthopaedic

Orthopaedic

Orthopaedic

Orthopaedic

Number

0/

patients

52

214

90

100

98

40

50

Control

23

18

14

16

66

50

48

Treated

o

5

I

2

50

50

40

230 A. G. G. TURPIE

with the placebo. On the other hand, studies by Hansen, Jessing, Lindewald,

Ostergaard, Olesen & Malver (1976), Hume et al. (1977), and Cooke, Dawson,

Ibbotson, Bowcock , Ainsworth & Pileher (1977) failed to demonstrate any benefit in

patients having surgery for fractured hip or after elective hip replacement using leg

scanning and venography to diagnose venous thrombosis.

Sulphinpyrazone

There have been two prospective trials on the use of sulphinpyrazone for the

prevention ofpostoperative deep vein thrombosis. Gruber, Fuser, Frick, Loosli, Matt

& Segesser (1977) reported the results ofa prospective study comparing the effects of

sulphinpyrazone, heparin and dextran on the incidence of venous thrombosis after

elective general surgery using leg scanning to detect venous thrombosis. Both dextran

and heparin were more effective than sulphinpyrazone in reducing the frequency of

postoperative venous thrombosis. Rogers, Walsh, Marder, Basak, Lachman, Ritchie,

Oppenheimer & Sherry (1978) carried out a randomized double-blind study

comparing sulphinpyrazone, low-dose heparin or placebo in patients undergoing hip

surgery and reported no significant reduction in the frequency of venous thrombosis

detected by leg scanning and venography in either treatment group compared with

placebo.

Conclusions

Of the platelet function suppressant drugs tested for the prevention of deep vein

thrombosis, aspirin is the only one that has been used in practice. However, the

results of the studies using aspirin have been inconsistent, although positive results

have been reported in patients undergoing orthopaedic procedures. Hydroxychloroquine has been demonstrated to reduce the frequency of deep vein thrombosis in

three studies of patients undergoing general abdominothoracic surgery but to be

ineffective in patients undergoing orthopaedic procedures. Dipyridamole and

sulphinpyrazone have been shown to be ineffective for the prevention of deep vein

thrombosis.

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