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Mucopolysaccharide polysulfate (MPS) represents a mammalian-derived sulfated polysaccharide. Because the origin and structure of heparins is similar to MPS, this study was conducted to compare 2 ointment formulations containing MPS or heparin with a placebo ointment on tissue factor pathway inhibitor (TFPI) released in nonhuman primates (Macaca mulatta). A primate colony composed of 18 animals, housed at Loyola University Medical Center, was used in compliance with an Institutional Animal Care and Use Committee (IACUC)-approved protocol. Mucopolysaccharide polysulfate (4.5%), heparin (4.5%), and a placebo ointment were topically applied to individual groups of primates in a crossover study for periods of up to 2 weeks. Blood samples were drawn on days 1, 2, 5, 7, and 10. The anticoagulant effects (activated partial thromboplastin time [APTT], Heptest, thrombin time [TT]), TFPI antigen and functional levels, thrombin activatable fibrinolytic inhibitor (TAFI), and antiheparin platelet factor 4 antibodies (AHPF4 abs) were measured in citrated plasma. All data were compiled as mean +/- 1 standard deviation and compared in groups. Topical administration of both the MPS and heparin ointments resulted in no measurable anticoagulant effects in the primate model; however, MPS produced a concentration-dependent release of TFPI antigen and a functional activity that was stronger than the effects observed with heparin. A decrease in TAFI activation was also observed in the MPS-treated primates. In addition, in the heparin-treated group, a slight increase in AHPF4 abs was observed. In conclusion, MPS showed a stronger release of TFPI than heparin that was not associated with a strong anticoagulant effect. Moreover, MPS downregulated TAFI, resulting in an enhanced fibrinolytic effect.

Superficial thrombophlebitis is a frequently occurring disease which, because of risk factors, affects women to a greater extent than men. The presence of deep venous thrombosis should be excluded by imaging procedures. Heparin or low-molecular weight heparins are indicated for treatment, if deep venous thrombosis is present or threatening. In the acute stage non-steroidal antiinflammatory drugs can be taken orally in order to alleviate pain and inflammation. In the case of concomitant chronic venous insufficiency, edema protective drugs (e. g. preparations containing horse chestnut extract and flavonoids) can support the therapy. For the treatment of inflammatory conditions in relatively short superficial vein segments without involvement of the deep venous system, a limitation to topical treatment is possible. In addition to compression therapy, the application of creams or gels containing heparin or heparinoids are the usual treatment approach. The use of topical products containing mucopolysaccharide polysulfate (MPS) has been proven to be beneficial. The problem of providing evidence of the efficacy of topical products in the therapy for superficial thrombophlebitis under the current strict demands of our regulatory authorities and of developing new products under these conditions is discussed.

Aprosulate sodium was the first representative of a new class of synthetic polyanions showing antithrombotic efficacy in different animal models. In several clinical trails (Phase I) in human volunteers aprosulate demonstrated anticoagulant properties, too. Searching for other active substances of similar structure, a series of bis-aldonic acid amides were synthesized. These compounds exhibited interesting antithrombotic and anticoagulant activities. The pharmacodynamic activities of the compounds LW 10121, LW 10125, LW 10114, 10244, and LW 10168 are summarized in this article. These substances prolonged the APTT to 150% of the blank values at concentrations of 1.5 to 13.5 micrograms/mL. The thrombin time and anti-Xa test were only slightly influenced by concentrations up to 100 micrograms/mL. All compounds were investigated in a jugular vein hemostasis model in rats to examine their antithrombotic potential. They all had an antithrombotic activity lower than aprosulate, except compound LW 10121, which seemed to be a little more active. The subcutaneous injection of 10 mg/kg LW 10121 resulted in a longer duration of action than aprosulate and heparin. On the basis of the chemical structure and the profile of action, it is assumed that the new compounds may possess the same mode of action as aprosulate, but the mechanism of action may be different from heparin and low molecular weight heparins.

A series of sulfated bis-lactobionic acid amides was prepared. The compounds comprise highly charged poly-anions of very low molecular weight. Usually, the compounds contain 16 sulfate groups per molecule and are homogeneous, monodisperse substances. The molecular weights range from 2388 to 2514. The compounds were evaluated for anticoagulant activity using a number of standard tests. The compounds exhibited moderate to good APTT activity. Interestingly, the anti-Xa and anti-IIa activities were very low, particularly in amidolytic assays. Using prothrombin and Factor X activation assays, it was demonstrated that these compounds were more potent in these experimental settings than in standard anti-Xa or anti-IIa assays. Compared with heparin and LMW heparins, the bis-lactobionic acid amides were particularly active as inhibitors of the intrinsic system activation of Factor X. The bis-lactobionic acid amides exhibited potent anticomplement activity, being clearly superior to heparin and LMW heparins. For the most interesting substance, LW 10082, experimental results suggest that its anticoagulant activity is independent of antithrombin II. The anticoagulant activity of LW 10082 was readily neutralized by protamine sulfate. Platelet factor 4, however, did not reduce its anticoagulant activity. Because of these favorable properties, it is hoped that LW 10082 might prove to be an interesting alternative to heparin or LMW heparin in clinical use.

1. The introduced bis-lactobionic acid amides represent a new group of synthetic polyanions with a molecular weight of 2387 to 2514d. The compounds are homogeneous and chemically defined. 2. The in vitro blood coagulation parameters (APTT, Heptest, anti-Xa, thrombin time, and anti-IIa) are only marginally altered by these compounds. 3. In a rat model of venous thrombosis using endothelial damage as the thrombogenic stimulus, the antithrombotic activity of the bis-lactobionic acid amides was shown in a dose- and time-dependent manner. 4. LW 10082 and LW 10121 prolong the bleeding time at dosages about 10 times less than that of heparin. The safety/efficacy index appears to be wider for these compounds than for heparin. 5. In subchronic toxicologic studies in rats, LW 10082 was well tolerated up to a dosage of 100 mg/kg body weight. 6. The anticoagulant and antithrombotic activities of the bis-lactobionic acid amides decrease gradually with the increasing number of methylene groups in the alkylene chain.

The cutaneous penetration of mucopolysaccharide polysulfate (MPS) from a commercial available combination drug (Mobilat) could be shown by means of histochemical and immunohistological methods. Deposition of MPS from ointment and gel in corium and subcutis of different animal species has been improved by the keratolytic activity of the salicylic acid component. There are differences in the intensity of absorption of MPS not only in the various animal species, but also caused by the particular galenic preparations investigated.

1. Phagocytic activity measured by means of the intravasal clearance of a soot dispersion in male NMRI-mice was increased six to ten days after whole-body X-irradiation (640 R) and decreased during the same period after i.v. administration of 2,4,6-triethyleneimino-s-triazine (TEM 2.0 mg/kg). 2. By means of 6-methyl-uracil food admixtures (200 to 400 ppm during 2 or 3 weeks) or by repeated intravenous injections of a N-methyl-D-glucosamine-6-methyluracil complex (62.5 to 250 mg/kg daily during five days), a significant augmentation of the phagocytic index being related to time and dosage was obtained in otherwise untreated mice. Comparable results were seen using cytidine and cytidine-s'-phosphate, whereas guanosine-5'-phosphate remained ineffective. 3. Whilst stimulating effects of 6-methyl-uracil or its N-methyl-D-glucosamine complex on X-irradiated mice were suspended, an increase up to supernormal values of the phagocytic index was produced by the pyrimidine base in animals treated with TEM. In accordance to this the survival rate of lethally X-irradiated mice (960 R) could not be increased; with animals given lethal TEM-doses, however, a significantly increased survival rate was obtained. 4. The present investigations as well as former biochemical analyses confirm the assumption that 6-methyluracil produces its regeneration effects, to some extent at least, by specific pathways influencing the reticuloendothelium. Different results from X-irradiated and TEM-treated mice are referring to the different points of attack of the two noxa.

The in vitro incorporation of 14C-thymidine into DNA of bone-marrow cells as well as DNA content of spleen and thymus of male Wistar rats were increased significantly by 6-methyluracil (within an N-methyl-D-glucosamine-6-methyluracil complex) and/or heterologous, low-molecular nucleic acids, when given therapeutically at doses of 100--250 mg/kg b.w. following i.v. injection of tretamine. Maximum effects were reached as early as two days after the onset of experiment. When tretamine was replaced by X-irradiation or cyclophosphamide, the influence of the 6-methyluracil complex and nucleic acids was less significant. Whereas normal bone-marrow cells and cells damaged in vitro by X-irradiation, tretamine and/or inhibitors of protein synthesis could not be influenced by the 6-methyluracil complex, incorporation of 14C-L-phenylalanine into proteins of bone-marrow cells was enhanced by exogenous DNA or RNA (10--100 micrograms/ml). The present investigations underline the previously drawn conclusions that certain biological alkylating agents, nucleic acids and pyrimidines may interfere to some extent through the reticuloendothelial system.

Survival time of mice after i.v. injections of 2,4.6-triethylene-imino-1,3,5-trazine (TEM) or total body-X-irradiation (TBI) was increased by 6-methyluraclil (6-MU) when given in food (200 ppm). Under the same conditions, 6-MU decreased the involution of spleen and thymus (as measured by DNA-content and DNase II activity) under the infuence of TEM and enhanced the regeneration of the spleen after TBI. Elevation of DNase I- and protein content of the kidneys and a (short-dated) increase of incorporation of 14C-phenylalamine into microsomes of liver of 14C-orotic acid into RNA of liver and kidney suggest that the influence of 6-MU is mediated at least partly by a specfically anabolic effect.

Antithrombotic activity of mucopolysaccharide polysulfuric acid ester (MPS) was investigated in vitro as well as in animal models of thrombosis. In vitro studies were performed by using native human blood and the haemostatometer. MPS (greater than or equal to 25 micrograms/ml) caused concentration dependent inhibition of the in vitro haemostasis (platelet reactivity). Antihaemostatic activity of epoprostenol (prostacyclin) was synergized by MPS. In vivo platelet thrombus formation was induced by laser irradiation in the microvasculature of hamster cheek pouch. The time until occlusion of the irradiated small vein and the duration of occlusion were measured. MPS administered intravenously at a dose range of 1-10 mg/kg b.w. inhibited thrombus formation and prolonged the time until occlusion of the irradiated vessel. Upon topical administration of a solution of 1, 5 or 25% MPS the occlusion time was also prolonged. Topical application of MPS as a 0.3% cream (Hirudoid) increased the occlusion time significantly. 24 h percutaneous application of 0.3% MPS cream but not the placebo on the back skin of rats resulted in a significant (p less than 0.001) inhibition of thrombus formation in the microvessels of the mesoappendix. A common finding at all modes of administration of MPS was that reperfusion of the occluded vessel occurred much earlier in MPS treated animals than in the controls.

It could be demonstrated by means of the UV erythema model that four different ointment preparations containing mucopolysaccharide polysulfate (MPS) have a marked antiinflammatory effect and inhibit the formation of erythemas to a large extent. In this trial arrangement the different preparations containing MPS were significantly superior to altogether eight commercial heparin ointments, which were tested, too. With an inhibition of 69% of the erythema induced by UV rays, Hirudoid 40000 ointment is the most effective of all preparations tested.

The anti-exudative and anti-edematous effects and the dose-effect relationship of a-sympathomimetics, especially of l-phenylephrine-HCl (PE), have been demonstrated using as model the rat pad-carrageenan edema and the histamine liberator test, the dosage being administered cutaneously, orally, and intraperitoneally. The beta-receptor activating compound bamethane sulfate was not effective when used on the same models. Evidence of percutaneous penetration of PE was provided both on isolated skin and in viro. PE, when injected intravenously or subcutaneously, produced a rise of blood pressure in experimental animals. When PE was administered orally and cutaneously, it did not, however, alter the blood pressure. The anti-inflammatory and anti-exudative effects remianed unchanged.