Antiplatelet action of ilexoside D,a triterpenoid saponin fromIlex pubescens

The anti-platelet activity of ilexoside D isolated from the roots ofIlex pubescens Hook. et Arn. was investigated inin vitro andex vivo models of platelet aggregation induced by ADP, thrombin or collagen in rats.In vitro ilexoside D inhibited more effectively platelet aggregation induced by ADP and thrombin than by collagen as compared with aspirin.Ex vivo ilexoside D also inhibited platelet aggregation induced by ADP and collagen, but not by thrombin, and the inhibitory action of ilexoside D was more effective than that of aspirin. However,in vitro ilexoside D inhibited very poorly the generation of malonyldialdehyde, which is known to be concomitantly released with thromboxane A₂ during platelet aggregation. These results suggest that the anti-platelet activity of ilexoside D may not be responsible for prostaglandin synthesis in platelets.     

Inhibition of Platelet Thromboxane Formation and Phosphoinositides Breakdown by Diisoeugenol

Abstract— Diisoeugenol inhibited the platelet aggregation and ATP release of rabbit platelets caused by ADP, arachidonic acid, platelet-activating factor (PAF), collagen and thrombin. Prolongation of the incubation time of platelets with diisoeugenol did not cause further inhibition and the aggregability of platelets could not be restored after washing. In human platelet-rich plasma, diisoeugenol inhibited the biphasic aggregation and ATP release induced by adrenaline and ADP in a concentration-dependent manner. Thromboxane B₂ formation caused by arachidonic acid, collagen and thrombin was markedly inhibited by diisoeugenol in a concentration-dependent manner. Diisoeugenol also inhibited the formation of inositol monophosphate caused by collagen, PAF and thrombin. The cAMP level of washed platelets was not changed by diisoeugenol. It is concluded that the antiplatelet effect of diisoeugenol is due to the inhibition of thromboxane formation and phosphoinositides breakdown.     

Mechanisms of petroleum hydrocarbon toxicity: studies on the response of rat liver mitochondria to Prudhoe Bay crude oil and its aliphatic, aromatic and heterocyclic fractions

The effect of Prudhoe Bay crude oil (PBCO) and its different fractions [aliphatic, aromatic, heterocyclic (NOS)] on the bioenergetic functions of isolated rat liver mitochondria were studied. A DMSO extract of PBCO inhibited state 3 respiration (in the presence of ADP) with either succinate or beta-hydroxybutyrate as substrate. The ascorbate-TMPD dependent state 3 respiration was not affected. Succinate dehydrogenase and beta-hydroxybutyrate dehydrogenase activities were also lost in the presence of the PBCO extract suggesting that inhibition of state 3 respiration may be due to blockage of the electron transport chain. Stimulation of state 4 respiration (in the absence of ADP) and of the oligomycin sensitive ATPase activity by the PBCO extract was observed. Fractionation of PBCO indicated that the aromatic fraction was mainly responsible for its inhibitory effects. By comparison, the heterocyclic fraction had weak inhibitory properties while the aliphatic fraction was essentially inactive. It is concluded that the aromatic components of PBCO inhibit mitochondrial respiration and oxidative phosphorylation mainly through impairment of the mitochondrial membrane and inhibition of beta-hydroxybutyrate and succinate dehydrogenase supported electron transfer activities of the respiratory chain.     

The effects of bisantrene on human platelets

Summary Bisantrene, (9,10-anthracenedicarboxaldehyde bis [(4,5-dihydro-lH-imidazol-2yl)hydrazone] dihydrochloride) is one of a series of anthracene dicarboxaldehyde compounds in Phase II trials. Preliminary studies suggested that bisantrene has anti-platelet activity. Therefore, in vitro studies of its effects on platelets were undertaken. Bisantrene in clinically attainable concentrations of 0.625 – 10 M, caused a 95 ± 1% decrease in maximal platelet aggregation to collagen (1 g/ml), and epinephrine (5–40 M) and 90 ± 10% inhibition of arachidonic acid (50 g/ml) induced aggregation. Collagen induced platelet shape change was not affected. Aggregation to calcium ionophore A23187 was inhibited by 10–30%. No effect on ADP induced aggregation was seen at clinically relevant bisantrene concentrations. This inhibition was time dependent, reaching a maximum when platelets were preincubated with bisantrene for 10 minutes before exposure to agonist. Inhibition persisted after bisantrene was removed by washing. To determine the mechanism of platelet inhibition, platelet prostaglandin metabolism, oxygen consumption and release reaction were measured. In the presence of drug: 1. normal cyclooxygenase activity was demonstrated by O₂ consumption studies and normal secondary wave ADP (3.2 M) aggregation; 2. lipoxygenase activity, by O₂ consumption was also normal. 3. There was 30–50% inhibition of thromboxane A2 synthesis induced by arachidonic acid or collagen; 4. ADP, collagen and AA induced release of serotonin was decreased by 30–60% but was never abolished; 5. No effect on basal platelet cAMP levels nor additive effect on PGE₁ induced elevation of platelet cAMP was detectable. These data demonstrate that bisantrene has potent antiplatelet activity probably mediated by several different mechanisms. The inhibition may have important clinical and theoretical consequences.Abbreviations PGl ₂ prostaglandin I₂ - PGEi prostaglandin E₁ - PRP platelet rich plasma - PPP platelet poor plasma - ACD acid citrate dextrose - TBX A₂ thromboxane Az₂ - TBX B₂ thromboxane B₂ - PAF platelet activating factor-acetyl glyceryl ether phosphoryl choline     

Antiplatelet Effects of Some Aporphine and Phenanthrene Alkaloids in Rabbits and Man

Two aporphines (boldine and laurolitsine) and five phenanthrene alkaloids (litebamine, secoboldine, N-cyanosecoboldine, N-methylsecoglaucine and N-methylsecopredicentrine) were evaluated in-vitro for their ability to inhibit platelet aggregation. All seven alkaloids inhibited aggregation of rabbit platelets and inhibited the release of ATP induced by arachidonic acid and collagen in rabbit platelets. Those aggregations induced by platelet-activating factor (PAF), thrombin, U46619 and ADP were inhibited by the three N-substituted secoboldine derivatives only. Thromboxane B₂ formation caused by arachidonic acid was also suppressed by these compounds. They did not affect the generation of [³H]inositol monophosphate caused by collagen, PAF and thrombin in the presence of indomethacin. Platelet cyclic AMP level was unaffected by litebamine, but was increased by N-methyl-secoglaucine. Litebamine suppressed the secondary aggregation, but not the primary aggregation, induced by ADP and adrenaline in platelet-rich plasma from man, whereas N-methylsecoglaucine inhibited both primary and secondary aggregation. It is concluded that the antiplatelet effect of these seven aporphine and phenanthrene alkaloids is mainly a result of inhibition of thromboxane A₂ formation; N-methylsecoglaucine has additional antiplatelet activity as a result of increasing the levels of platelet cyclic AMP.     

Studies on the mechanism of action of 12-deoxyphorbolphenylacetate,a potent platelet aggregating tigliane ester

12-Deoxyphorbolphenylacetate (12-DOPP) induced human platelet aggregation which was dependent upon the presence of divalent cations, the intracellular level of cyclic-AMP and an intact microtubular system, in common with other aggregating agents. However platelet secretion and thromboxane (Tx)B₂ synthesis did not contribute to 12-DOPP induced platelet aggregation as neither the Tx/endoperoxide antagonists pinane TxA₂ and trimethoquinone, the thromboxane synthetase inhibitors clotrimazole and 9, 11, aza-prosta-5-13-dienoic acid nor the cyclo-oxygenase inhibitor indomethacin inhibited 12-DOPP induced aggregation. Furthermore the free radical scavengers aminopyrine, thioanisole and butylated hydroxy-toluene, the lipoxygenase inhibitor phenidone and the leucotrienes B and C antagonist FPL 55712 failed to modify 12-DOPP-induced aggregation.Compounds which are thought to act as phospholipase inhibitors (bromophenacylbromide, mepacrine and propranolol as well as imipramine, desmethylimipramine, promethazine and trifluoperazine) which have been shown to selectively bind calmodulin, were found to be effective inhibitors of 12-DOPP induced aggregation. Aggregation induced by 12-DOPP involves a direct effect upon platelets followed by the release of unknown substances, probably phospholipid products. The released substances were shown to induce further aggregation of platelets. The aggregation induced by 12-DOPP is thus distinct from that induced by ADP, collagen, adrenaline and prostaglandin endoperoxides.     

Cardiotoxin from Naja naja atra snake venom: A potentiator of platelet aggregation

Cardiotoxin, isolated from Naja naja atra snake venom, potentiates platelet aggregation induced by ADP, thrombin, collagen and venom phospholipase A₂. The malondialdehyde formation caused by ADP, thrombin and venom phospholipase A₂ were also increased in the presence of cardiotoxin. Both potentiation of aggregation and increase in malondialdehyde were blocked by indomethacin or Ca²⁺ (5 mM or 0.05 mM). Cardiotoxin did not potentiate thrombin-induced aggregation of p-bromophenacyl bromide-modified platelets. Thromboxane B₂ formation induced by thrombin or collagen was also increased by cardiotoxin, while that by arachidonate was not affected. As a membrane-active polypeptide, cardiotoxin might augment the Ca²⁺-flux during the activation of the platelet membrane by aggregation inducers and then increase the activation of endogenous phospholipase A₂.     

Antiplatelet Effect of Gingerol Isolated from Zingiber officinale

The purpose of this investigation was to determine the antiplatelet mechanism of gingerol. Gingerol concentration-dependently (0·5–20 μm ) inhibited the aggregation and release reaction of rabbit washed platelets induced by arachidonic acid and collagen, but not those induced by platelet-activating factor (PAF), U46619 (9,11-dideoxy-9α,11 α-methano-epoxy-PGF_2α) and thrombin. Gingerol also concentration-dependently (0·5–10μ m ) inhibited thromboxane B₂ and prostaglandin D₂ formation caused by arachidonic acid, and completely abolished phosphoinositide breakdown induced by arachidonic acid but had no effect on that of collagen, PAF or thrombin even at concentrations as high as 300 μ m . In human platelet-rich plasma, gingerol and indomethacin prevented the secondary aggregation and blocked ATP release from platelets induced by adenosine 5′-diphosphate (ADP, 5 μ m ) and adrenaline (5 ä m ) but had no influence on the primary aggregation. The maximal antiplatelet effect was obtained when platelets were incubated with gingerol for 30 min and this inhibition was reversible. It is concluded that the antiplatelet action of gingerol is mainly due to the inhibition of thromboxane formation.     

Comparison of the inhibitory effects of some compounds present in crude oils on rat platelet aggregation: role of intra- and extra- cellular calcium

In vitro addition of some representative aliphatic, aromatic or heterocyclic compounds present in petroleum crude oils to washed rat platelets resulted in a concentration-dependent inhibition of aggregation induced by ADP or thrombin. Increasing concentration of extracellular Ca2+ did not alter the pattern of inhibition. ADP-induced intracellular Ca2+ mobilization was unaffected by most of the compounds tested. However, Ca2+ uptake was significantly inhibited when platelets were preincubated with these agents. This suggests that some components of crude oil may inhibit platelet aggregation by bringing about alterations in the platelet plasma membrane.     

Effects of reserpine on rabbit platelet aggregation and adherence to collagen or injured rabbit aorta

Effects of reserpine in vivo and in vitro on rabbit platelets in citrated platelet-rich plasma and in suspensions of washed platelets have been studied. Administration of reserpine ($\text{5}\text{mg}\text{kg}$) intraperitoneally 18 hr before platelets were isolated caused inhibition of collagen-induced aggregation but not of aggregation induced by ADP or thrombin. Thrombin-induced aggregation was slightly enhanced. Platelets from reserpine-treated rabbits were less adherent than control platelets to collagen-coated glass surfaces or to the subendothelium of the rabbit thoracic aorta. Similar effects on aggregation were obtained when reserpine (0.2 to 10 μM) was added to suspensions of washed rabbit platelets as little as 2 sec before the addition of collagen. Collagen-induced release of nucleotides and [¹⁴C]serotonin from prelabeled washed rabbit platelets was not affected by the presence of reserpine, whereas thrombin-induced release was slightly enhanced. Inhibition by reserpine (2–10 μM) of platelet adherence to a collagen-coated surface or to the subendothelium was also observed within a time interval too short for the reserpine to have caused depletion of platelet granule contents. Thus, reserpine has an immediate effect on the plasma membrane of the platelets which is responsible for inhibition of platelet adherence to collagen and hence of collagen-induced aggregation. This inhibitory effect differs from a much slower effect of reserpine at the granule membrane which results in the depletion of the granule contents of serotonin and adenine nucleotides. The effect of reserpine is not abolished by washing and resuspending platelets that have been exposed to reserpine in vivo. By inhibiting the interaction of platelets with collagen, reserpine may interfere with one of the components of hemostatic plug and thrombus formation.     

Serum albumin enhances the impairment of platelet aggregation with thromboxane synthase inhibition by increasing the formation of prostaglandin D2

Dazoxiben, a thromboxane synthase inhibitor, inhibits arachidonic acid induced aggregation in platelet-rich plasma from some donors only (“responders”). We have studied the effect of dazoxiben in vitro on platelet aggregation and prostaglandin (PG) metabolism and the influence of the incubation period and of exogenously added serum albumin (SA). SA, which increases the production of anti-aggregatory PGD₂ from cyclic endoperoxides, induced “non-responder” human platelets to respond. With rabbit platelets, however, that are insensitive to PGD₂, exogenous SA failed to potentiate dazoxiben-induced inhibition. The ratio between PGD₂ and TXB₂ + PGE₂ formed was crucial in determining the response of human platelets to dazoxiben: whenever this ratio was high, platelet aggregation was inhibited. SQ 22536, an adenylate cyclase inhibitor, and NO164, a PGD₂ antagonist, reversed the inhibition by dazoxiben in human platelet-rich plasma, stressing the importance of a PGD₂ mediated rise of cyclic AMP for the effectiveness of a thromboxane synthase inhibitor.     

A brief review on high on-aspirin residual platelet reactivity

Although aspirin is effective in secondary prevention in coronary heart disease, new thromboembolic events in patients on aspirin are frequently seen. In trials on aspirin-treated patients, platelet function tests have revealed large variability in platelet aggregation. This phenomenon has been named aspirin resistance, aspirin non-responsiveness or high-on-aspirin residual platelet reactivity.The mechanism of aspirin antiplatelet effect is due to the inhibition of cyclooxygenase-1 enzyme in platelets. In some trials, almost all patients on aspirin have a very low level of serum thromboxane B₂, indicating that the measured platelet reactivity in aspirin-treated patients might be due to platelet activation via other pathways, such as ADP or thrombin. The prevalence of real aspirin resistance seems to be very low, and probably the term “high-on-aspirin residual platelet reactivity” should be preferred to describe this phenomenon.     

Prostaglandin endoperoxides, thromboxane A2 and adenosine diphosphate in collagen-induced aggregation of rabbit platelets

1 A bioassay technique is described for simultaneously monitoring rabbit platelet aggregation with measurement of thromboxane A₂ (TxA₂) and prostaglandins released in response to collagen or arachidonic acid (AA).

2 Five imidazole derivatives were examined as inhibitors of thromboxane synthetase and compared with the effect of the cyclo-oxygenase inhibitor indomethacin; 1-(7-carboxyheptyl) imidazole was identified as the most potent and selective inhibitor of thromboxane synthetase and was used with indomethacin to investigate the relative contribution of the prostaglandin endoperoxides prostaglandin G₂ (PGG₂)/PGH₂ and TxA₂ in mediating platelet aggregation induced by collagen or AA.

3 Platelet aggregation induced by a low concentration of collagen was abolished by indomethacin and carboxyheptylimidazole whilst in response to a high concentration or collagen only partial inhibition of aggregation occurred.

4 The contribution of adenosine diphosphate (ADP) released from platelets during collagen or AA-induced aggregation was examined using the substrate/enzyme complex creatine phosphate/creatine phosphokinase (CP/CPK). The CP/CPK complex abolished aggregation induced by a low dose of collagen whilst aggregation to a high dose of collagen was only partially inhibited.

5 Aggregation induced by a high dose of collagen was abolished by a combination of CP/CPK with indomethacin or carboxyheptylimidazole.

6 AA-induced aggregation was abolished by indomethacin. Carboxyheptylimidazole abolished aggregation induced by a low dose of AA but inhibition was surmounted with increasing concentrations of AA in the absence of TxA₂ formation.

7 PGH₂-induced aggregation was unaffected by indomethacin and only partially inhibited by carboxyheptylimidazole. AA or PGH₂-induced platelet aggregation was unaffected by CP/CPK.

8 In conclusion, aggregation of rabbit platelets induced by a low concentration of collagen was dependent on synergism between TxA₂ and ADP whilst at high concentrations of collagen, sufficient TxA₂ and ADP were released to induce aggregation independently of each other.

9 The small amounts of prostaglandin endoperoxides produced from endogenous arachidonate have apparently no direct pro-aggregatory role. However, the relatively large amount which can be produced by a high concentration of exogenous AA when TxA₂ formation is prevented can cause aggregation of rabbit platelets.

     

Dicentrine, a novel antiplatelet agent inhibiting thromboxane formation and increasing the cyclic AMP level of rabbit platelets.

Dicentrine is an antiplatelet agent isolated from the Chinese herb Lindera megaphylla. We examined the in vitro effects of dicentrine on various aspects of platelet reactivity. Dicentrine inhibited the aggregation and ATP release of washed rabbit platelets induced by arachidonic acid (AA), collagen, ADP, platelet-activating factor (PAF), thrombin and U46619. Dicentrine also inhibited the thromboxane B2 formation caused by AA, collagen and thrombin in washed intact platelets or that induced by AA in lysed platelet homogenate, while prostaglandin D2 formation caused by AA was not increased. The generation of inositol monophosphates (in the presence of indomethacin) caused by thrombin, collagen and PAF was not suppressed significantly, nor did dicentrine suppress fibrinogen-induced aggregation of elastase-treated platelets. Dicentrine inhibited the intracellular Ca2+ increase in quin-2/AM-loaded platelets caused by thrombin, PAF, collagen and AA. The cyclic AMP level was elevated by dicentrine in a concentration-dependent manner. These data indicate that the inhibitory effect of dicentrine on platelet aggregation and ATP release was due to the inhibition of thromboxane formation and the elevation of the level of cyclic AMP.     

“In vitro” and “ex vivo” effects of picotamide,a combined thromboxane A2-synthase inhibitor and -receptor antagonist,on human platelets

Summary Picotamide (G 137), a new non prostanoid inhibitor of in vitro arachidonic acid induced platelet aggregation, has been further characterized in in vitro and ex vivo studies.When whole blood was activated with collagen in the presence of picotamide 5×10^–4 M, thromboxane B₂ production was decreased, and 6-keto-PGF₁ generation was significantly increased, suggesting a reorientation of platelet endoperoxide metabolism following blockade of thromboxane synthetase. Picotamide also inhibited platelet aggregation and clot retraction induced by the endoperoxide analogue U46619 in human platelets, indicating thromboxane A₂-receptor antagonism, possibly of competitive nature.A single oral dose of picotamide 1 g in 24 healthy volunteers produced a significant inhibition of collagen, arachidonic acid and U46619-induced platelet aggregation. Serum levels of thromboxane B₂ were also reduced.Chronic administration of picotamide 1.2 g/d to patients with vascular disease resulted in a prompt and persistent fall in their increased plasma levels of -thromboglobulin.The results indicate that picotamide is a combined thromboxane B₂-synthetase inhibitor and thromboxane A₂-receptor antagonist in human platelets, and that it may prove useful as an antithrombotic agent.     

莽草酸对血小板聚集和凝血的抑制作用

目的：研究莽草酸(SA)对血小板聚集性的影响及其机制,并研究其对凝血时间的影响。 方法：采用比浊法测定血小板聚集功能,放射免疫法测定血小板TXB2和血浆6-keto-PGF_1α水平,玻管法测定凝血时间。结果：SA体外呈浓度依赖性抑制ADP、胶原诱导的兔血小板聚集,IC₅₀分别为9.25, 3.56 mmol.L^-1;在体内(ip)抑制脑血栓模型鼠由ADP诱导的血小板聚集; SA促进大鼠血浆6-keto-PGF_1α的生成并延长小鼠凝血时间。结论：SA对血小板聚集及凝血系统有显著的抑制作用。     

Effects of compounds extracted from Santolina oblongifolia on TXB2 release in human platelets

A group of compounds isolated from the medicinal plant, Santolina oblongifolia, have been investigated for their effects on the release of platelet cyclo-oxygenase metabolite thromboxane A₂ (measured as thromboxane B₂) from ionophore-stimulated human platelets. These compounds, which are dual inhibitors of cyclo-oxygenase (prostaglandin E₂) and lipoxygenase (leukotriene C₄) activity in vitro, are: apigenin, luteolin, quercetin, herniarin, scopoletin, scopolin and aesculetin. All compounds assayed presented a dose-related response to thromboxane B₂ release, with the percentages of inhibition being slightly lower than the reference drug, ibuprofen. Inhibition was more evident with the flavonoids. Our data support the inhibition of arachidonic acid metabolism as one of the mechanisms for which flavonoids and coumarins from S. oblongifolia exert their anti-inflammatory effect     

Mechanism of Action of p-Chlorobiphenyl on the Inhibition of Platelet Aggregation

p-Chlorobiphenyl (1–50 μm ) concentration-dependently inhibited the aggregation and release reaction of rabbit washed platelets induced by arachidonic acid and collagen, but not those induced by platelet-activating factor (PAF), U46619 and thrombin. The IC50 values of p-chlorobiphenyl on the arachidonic acid and collagen-induced platelet aggregation were 2.9 ± 0.5 and 12.8 ± 2.3 μm , respectively. The formation of both platelet thromboxane B₂ and prostaglandin D₂ caused by arachidonic acid was inhibited by p-chlorobiphenyl concentration-dependently. In myo-[³H]inositol-labeled and fura-2-loaded platelets, [³H]inositol monophosphate generation and the rise in intracellular Ca²⁺ stimulated by arachidonic acid were inhibited by p-chlorobiphenyl. In human platelet-rich plasma, p-chlorobiphenyl and indomethacin prevented the secondary aggregation and blocked ATP release from platelets induced by adenosine 5′-diphosphate and adrenaline without affecting the primary aggregation. It is concluded that p-chlorobiphenyl may be a cyclo-oxygenase inhibitor and its antiplatelet action is mainly due to the inhibition of thromboxane formation.     

Effects of cryptolepine on collagen-induced aggregation and on the mobilization and metabolism of arachidonic acid by rabbit platelets

1. We examined the effect of cryptolepine on collagen-induced aggregation and on the mobilization, and metabolism of arachidonic acid in rabbit platelets.2. Preincubation of platelets with cryptolepine (50–100 μM) did not affect the primary wave of aggregation but resulted in a dose-dependent, surmountable inhibition of the secondary wave of aggregation induced by collagen (5 μg/ml). The inhibition by cryptolepine was greater when cryptolepine was incubated with the platelets after the peak of the primary wave of aggregation.3. Cryptolepine (50–100 μM) dose-dependently inhibited thrombin (1.5 U/ml) and A23187 (2.5 μM)-induced release of ¹⁴C[AA] from platelet membrane phospholipid pools. The percentage inhibition of A23187-induced ¹⁴C[AA] release was 31.3 ± 4.3% (50 μM) and 79.3 ± 5.4% (100 μM), while thrombin-induced release was inhibited by 39.2 ± 2.4% (50 μM) and 68.2 ± 3.6% (100 μM).4. At near maximal concentration (100 μM) which significantly inhibited secondary aggregatory response and ¹⁴C[AA] release, cryptolepine had no effect on the platelet metabolism of ¹⁴C[AA] to thromboxane B₂, HHT and 12 HETE.5. The present findings suggest that cryptolepine inhibited collagen-induced secondary aggregation through a selective antiphospholipase-like activity. There was not effect on platelet cyclooxygenase and lipoxygenase activities of platelets.     

Stereo-dependent inhibition of human platelet function by the optical isomers of trimetoquinol

The stereoisomers of trimetoquinol [1-(3',4',5'-trimethoxybenzyl)-6–7-dihydroxy-1,2,3,4-tetrahydroisoquinoline; TMQ] were shown to have potent and selective inhibitory effects on human platelet function in vitro. the R(+)-isomer was 12.1-, 12.3-, 39.2-, 82.9- and 36.0-fold more effective than the S(?)-isomer as an inhibitor of aggregation induced by arachidonic acid (AA), collagen, the epoxymethano-PGH2 analogs U44069 and U46619, and thromboxane A; (TxA₂) respectively. the concentrations of the R(+)-isomer that produced 50 percent inhibition (IC₅₀) of platelet aggregation were 4.2, 4.3, 1.4, 0.14 and 0.64 μM using AA, collagen, U44069, U46619, and TxA₂₀ as respective inducers. the graphical approximation of an inhibitory Constant (K_i = 0.13 μM) for the effect of TMQ on U46619-induced aggregation suggested that a competitive-like inhibition was operative. In other experiments, platelet aggregation and serotonin release induced by U46619 were inhibited differentially by the TMQ stereoisomers with nearly identical concentration-response curves. In addition, racemicTMQ blocked the secondary phase of platelet aggregation and serotonin release induced by ADP. These data, together with the ability of the TMQ stereoisomers to selectively inhibit TxA₂-induced aggregation, suggest that TMQ is an inhibitor of endoperoxide or TxA₂ action, e.g. a thromboxane A₂ receptor antagonist.