Methods and Compositions for Thrombolytic Therapy

In one set of experiments with no exogenous plasminogen, TPA was employed as the plasminogen activator at different levels with and without 200.mu.M cerulenin as the factor XIIIa inhibitor. The results seen in FIG. 1 show that there are enhanced rates and extents of lysis in the presence of cerulenin at both physiological and pharamcological levels of TPA.

In another set of experiments, 12.mu.g/mL of exogenous glu-plasminogen (glutamic plasminogen) was employed to enhance the rate and extent of TPA catalyzed clot lysis. Under these conditions in the presence of 200.mu.M cerulenin, lysis rates were further enhanced in a relative similar way as seen in FIG. 2.

In still another set of experiments with no exogenous plasminogen, TPA was employed as the plasminogen activator at different levels with and without the thiazolium perchlorate compound as the factor XIIIa inhibitor compound. The results seen in FIG. 3 show that there are enhanced rates and extents of lysis in the presence of the thiazolium perchlorate compound at both physiological and pharmacological levels of TPA.

In FIGS. 1 and 2, the open symbols indicate absence of cerulenin and the closed symbols indicate the presence of 200.mu.M cerulenin at the time of clot formation. Clot lysis is conducted in buffer, pH 7.5; 5 ng/mL TPA represents approximate physiologic concentration. 50 to 5,000 mg/mL represents pharmacologic concentration range. FIG. 1 studies did not employ exogenous glu-plasminogen. FIG. 2 studies employed glu-plasminogen levels at approximately 10 percent physiologic level (12.mu./mL)

The process of the present invention for enhancing clot lysis and/or for inhibiting clot formation comprises intravenously administering a therapeutic dose of a Factor XIIIa inhibitor compound in a composition comprising the same. In general, the dose may be that sufficient to provide between about 2 micrograms per kilogram of body weight per day to about 100 milligrams/kilogram/day while considering patient's health, weight, age and other factors which influence drug response. The drug may be administered either by a single injection, multiple injections or continuous infusion.

In the preferred process of the present invention, Factor XIIIa inhibitor compound is administered with a plasminogen activator in a combination therapy. When Factor XIIIa inhibitor compound and plasminogen activator are employed in a combination therapy, it is most desirable to use plasminogen activator in the dose range of 5 to 40,000 I.U./kg/hr and Factor XIIIa inhibitor compound in the range of 10 .mu.g - 10 mg/kg/hour.

When combination therapy is employed, it is preferable to administer the Factor XIIIa inhibitor compound first in a single bolus and thereafter to administer the plasminogen activator by continuous infusion. Alternatively, it may be administered simultaneously as a continuous infusate. Under certain circumstances it may be desirable to administer subsequent to the administration of the plasminogen activator. However, it is intended that the method of the present invention embrace concurrent administration as well as sequential administration in any order.

When the Factor XIIIa inhibitor compound is employed alone, particularly for a prophylactic purpose, it may be employed in the range of from 10 .mu.g to 100 mg/kg/day and administered either orally or parenterally.

Compositions to be employed in the practice of the present invention comprises a Factor XIIIa inhibitor compound in sterile physiologically acceptable media such as physiological saline. Such compositions may also contain other ingredients for purposes such as for aiding solubility or for preservation or the like, said ingredients being acceptable for intravenous administration. The compositions may be prepared as concentrate compositions which may be appropriately diluted to the appropriate treating composition immediately prior to administration. A therapeutic composition as a unitary dose form may contain from 2 .mu.g to 1000 mg of Factor XIIIa inhibitor compound and as concentrate composition may contain up to 10 grams of the compound. Compositions suitable in the preferred practice of the present invention of co-administering plasminogen activator and Factor XIIIa inhibitor compound may contain from 50,000 to 2 million I.U. of TPA and from 100 mg to 7 grams of Factor XIIIa inhibitor compound.

The preferred compositions are those in which the Factor XIIIa is cerulenin, 2-(1-acetonyl-thio)-5-methylthiazolo(2,3-b)-1,3,4-thiadiazolium perchlorate or 3-(benzimidazol-2-yl)acrylic acid.

Parenteral compositions may be prepared employing the foregoing preferred compounds or one of the following compounds as active ingredient in a manner hereinafter described. These compositions are merely illustrative and are not to be construed as limiting.

3-(1-oxo-7-nonaenyl)oxiranecarboxamide

3-(1-oxo-nonanyl)oxiranecarboxamide

3-(1-oxo-6-phenylpropyl)oxiranecarboxamide

N-Methyl-3-(1-oxo-4,7-nonadienyl)oxiranecarboxamide

n-Propyl 3-(1-oxo-4,7-nonadienyl)oxirancecarboxylate

3-(5-methylbenzimidazol-2-yl)acrylic acid

3-(5-isopropoxybenzimidazol-2-yl)acrylic acid

3-(5-chlorobenzimidazol-2-yl)acrylic acid

3-(Imidazol-2-yl)acrylic acid

3-(Oxazol-2-yl)acrylic acid

3-(Thiazol-2-yl)acrylic acid

3-(Benzoxazol-2-yl)acrylic acid

3-(Benzthiazol-2-yl)acrylic acid

One liter of a parenteral composition comprising one of the preferred compounds or one of the foregoing compounds may be prepared from the following formulation:

The parabens, sodium chloride and carboxymethylcellulose are dissolved in one-half the total volume of water by heating to 95.degree. C. to obtain a solution which is then filtered and autoclaved. The polysorbate is dissolved in one-third of the total volume of water, and the resulting solution also filtered and autoclaved. Sterile active ingredient is added to the second solution and the mixture passed through a sterile colloid mill to obtain a suspension of active ingredient. The first solution is added to the suspension with stirring then U.S.P. water added to 1 liter. Sterile vials are filled with the suspension while stirring.

Oral compositions also may be prepared from one of the above-named compounds as active ingredient in admixture with a pharmaceutically acceptable carrier. Suitable carriers include for liquid compositions include water, glycols, oils, alcohols, flavoring agents, preservatives, coloring agents and the like; for solid preparations, starches, sugars, diluents, granulating agents, lubricants, binders, disintegrating agents and the like. Because of their ease in administration, tablets and capsules represent the most advantageous oral dosage unit form, in which case solid pharmaceutical carriers are obviously employed.

A representative composition is the following:

5000 compressed tablets, each containing as active ingredient 100 milligrams of active ingredient are prepared from the following formulation:

The ingredients are finely powdered, mixed well, and then granulated with 10 percent starch paste. The granulation is dried and compressed into tablets using starch as a disintegrant and calcium stearate as lubricant.