Compositions and methods for selective dissolution of nascent intravascular blood clots

Abstract

Compositions and methods for prevention and treatment of uncontrolled formation of intravascular fibrin clots, which are capable of selective dissolution of pathological nascent clots formed intravascularly, with minimal risk of unwanted dissolution of pre-existing hemostatic clots, are provided wherein fibrinolytic or anticoagulant drugs are biocompatibly coupled to red blood cell carriers.

Description

[0001] This application is a Divisional Application of U.S. patent application Ser. No. 10 / 611,723 filed Jul. 1, 2003, which is a continuation-in-part of U.S. patent application Ser. No. 10 / 253,518 filed Sep. 23, 2002, which is a divisional application of U.S. patent application Ser. No. 09 / 454,666 filed Dec. 3, 1999, now issued as U.S. Pat. No. 6,488,927, which is a continuation-in-part application of PCT Application PCT / US1999 / 10547 filed May 12, 1999, which claims the benefit of priority to U.S. Provisional Application Ser. No. 60 / 086,262 filed May 21, 1998, each of which are herein incorporated by reference in their entirety.BACKGROUND OF THE INVENTION [0002] Occlusions of blood vessels by intravascular clots cause or / and contribute to the pathogenesis of a variety of disease conditions including myocardial infarction, stroke and pulmonary embolism and thus represent a significant medical problem. Although fibrinolytics, such as plasminogen activators, have recently been used in...

AI Technical Summary

Benefits of technology

"The present invention provides compositions that contain a therapeutic agent, such as an anti-thrombotic agent, that is attached to a carrier, such as a red blood cell. These compositions can circulate for a long time and specifically dissolve nascent blood clots in the blood vessels. The therapeutic agent is attached to the carrier through a cross-linking process using streptavidin or by using molecules with high affinity to specific proteins on the surface of the red blood cell. The invention also provides a method for preventing and treating uncontrolled formation of blood clots while avoiding lysis of pre-existing clots by administering the therapeutic agent-red blood cell composition to patients."

Problems solved by technology

Occlusions of blood vessels by intravascular clots cause or / and contribute to the pathogenesis of a variety of disease conditions including myocardial infarction, stroke and pulmonary embolism and thus represent a significant medical problem.

Pulmonary thromboembolism, a leading cause of mortality, is most often a complication of deep venous thrombosis.

Even with this well-known drug, however, there is no consensus regarding the optimum regimen of anticoagulant therapy that affords both safety and efficacy.

Acute vascular occlusion by fibrin thrombi is also a common and dangerous complication of surgery.

Development of venous (e.g., pulmonary embolism) and arterial (e.g., arterial re-occlusion) thrombi that may embolize vitally important blood vessels increases peri-operative morbidity and mortality.

Cardiopulmonary bypass surgery, transcutaneous coronary angioplasty and carotid endarterectomy and other interventions may be complicated by formation of small blood clots, which embolize to cerebral vessels and cause serious persistent neurological disorders.

In contrast to settings where ischemia is the sole threat to health (e.g., acute myocardial infarction, AMI), the management of thrombosis in the surgical settings is complicated by the added risk of bleeding at and around the wound site imparted by anti-thrombotic therapy itself.

In addition to the objective inadequacies of existing agents (see below), widespread use of thromboprophylaxis in surgical settings continues to receive limited enthusiasm for additional reasons.

Current approaches afford some measure of protection, but all have a limited benefit / risk ratio in the surgical setting.

Mechanical filters trap large clots, but cannot be used to prevent arterial re-occlusion or cerebral micro-emboli.

Anticoagulants (e.g. low molecular weight heparin) reduce the incidence of thrombosis in some vascular beds, but dosing is limited by the risk of post-operative bleeding.

Heparin has been reported to increase the incidence of wound hematomas, which in turn may predispose to wound infection and dehiscence.

Aspirin and anti-GPIIb / IIIa agents suppress platelet aggregation, but cannot prevent formation of fibrin monomers and clot propagation.

In addition to serious side effects (in part discussed above), each particular anticoagulant agent inhibits one specific mechanism for formation of clots (e.g., activation of platelets or fibrin formation), but none affords 100% fidelity of anti-thrombotic protection.

The risk of hemorrhage practically precludes their use in the surgical settings.

No existing anti-thrombotic strategy permits selective dissolution of the nascent clots without affecting pre-existing hemostatic fibrin plugs.

However, application of these agents to dissolution of clots formed or lodged in other vascular areas such as deep venous areas is limited by extremely rapid elimination and inactivation after bolus dosing (Plow, E. et al.

Such inactivation, or degradation of plasminogen activators and plasmin reduce the effectiveness of thrombolytic therapy and thus fail to prevent re-occlusion of blood vessels.

To overcome this problem, attempts have been made to infuse plasminogen activators intravenously for prolonged periods of time with little success; failure was attributed to the harmful side effects such as bleeding, dissolution of hemostatic clots as well as nascent clots causing deleterious consequences, and uncontrolled tissue proteolysis that occurred after extravascular deposition of plasminogen activators.

However, these studies have shown that the activity of liposome-encapsulated plasminogen activators is strongly compromised by steric limitations.

In addition, clots bind only a small fraction of injected “fibrin-specific” plasminogen activator because of limited surface area of the formed clots.

Such targeting also fails to distinguish nascent clots with deleterious consequences from pre-existing desired hemostatis clots.

Further, to date, none of these methods for modifying plasminogen activators prevent deposition of plasminogen activators in tissues, which can lead to an increase in harmful side effects; they all represent molecules or molecular complexes with sizes that do not exceed that of blood proteins.

Activated plasmin degrades the extracellular matrix, thus causing vascular remodeling, abnormal elevation of vascular permeability and even partial denudation of subendothelium (Plow et al.

However, polyvalent conjugation of biotinylated proteins to b-RBCs via streptavidin cross-linker profoundly compromises the biocompatibility of the carrier RBC.