Thrombolytic Science, LLC

Other

New England Deaconess Hospital Corporation

Beth Israel Deaconess Medical Center, Inc.

Ffe Therapeutics LLC

Victor Gurewich

Jian-ning Liu

Ralph Pannell

John Noel Williams

Paolo Sarmientos

Massimiliano Pagani

Boguslaw Lipinski

Syed S Huasin

Provided herein are methods for safe and effective thrombolysis in therapy for human subjects with symptoms of a potential stroke or acute myocardial infarction ('AMI') using a sequential administration of a low dose bolus of human tissue plasminogen activator ('tPA') followed by an infusion of a mutant form of human pro-urokinase (&#x201c;proUK&#x201d;).

Methods for safe and effective thrombolysis using sequential administration of tissue plasminogen activator and mutant pro-urokinase

Provided herein are methods for use in safe and effective thrombolysis, e.g., in therapy for a potential stroke or acute myocardial infarction ('AMI') at a maximum arterial patency rate with minimal associated hemorrhagic side effects.

Methods for thrombolysis

Provided herein are anti-angiogenic monoclonal antibodies and antigen-binding antibody fragments that selectively and specifically bind to an epitope in both fibronectin and either fibrinogen or fibrin fragment E, compositions containing these antibodies and antibody fragments, and methods of using these antibodies and antibody fragments.

Compositions and methods for treating angiogenesis-related disorders

A mutant prourokinase plasminogen activator (M5) was developed to make prouPA less subject to spontaneous activation during fibrinolysis. C1-inhibitor complexes with tcM5. The effect of C1-inhibitor on fibrinolysis and fibrinogenolysis by M5 was determined. Supplemental C1-inhibitor restores the stability of M5 but not that of prouPA. Clot lysis by M5 with supplemental C1-inhibitor showed no attenuation of the rate of fibrinolysis, whereas fibrinogenolysis was prevented by C1-inhibitor. Due to higher dose tolerance of M5 with C1-inhibitor, the rate of fibrin-specific lysis reached that achievable by nonspecific fibrinolysis without inhibitor. Plasma C1-inhibitor stabilized M5 in plasma by inhibiting tcM5 and thereby non-specific plasminogen activation. At the same time, fibrin-specific plasminogen activation remained unimpaired. This unusual dissociation of effects has significant implications for improving the safety and efficacy of fibrinolysis. Methods of reducing bleeding and non-specific plasminogen activation during fibrinolysis by administering M5 along with exogenous C1-inhibitor are disclosed.

C1-inhibitor prevents non-specific plasminogen activation by a prourokinase mutant without impeding fibrin-specific fibrinolysis

C-1 inhibitor prevents non-specific plasminogen activation by a prourokinase mutant without impeding fibrin-specific fibrinolysis

It has now been discovered that certain mutant forms of pro-urokinase ('pro-UK'), such as so-called pro-UK mutant 'M5' (Lys&#x2192;His), perform in the manner of pro-UK in lysing &#x201c;bad&#x201d; blood clots (those clots that occlude blood vessels), while sparing hemostatic fibrin in the so-called &#x201c;good&#x201d; blood clots (those clots that seal wounds, e.g., after surgery or other tissue injury). Thus, these pro-UK mutants are excellent and safe thrombolytic agents. These advantages allow them to be used in a variety of new methods, devices, and compositions useful for thrombolysis and treating various cardiovascular disorders in clinical situations where administration of other known thrombolytic agents has been too risky or even contraindicated.

Methods, devices, and compositions for lysis of occlusive blood clots while sparing wound sealing clots

A fusion drug including an isolated portion of the A-chain of a urokinase-type plasminogen activator linked to a drug, wherein the A-chain portion binds stably to an outer membrane of a platelet. The T.sub.1/2 of the fusion drug in plasma is thereby increased to about 4 to 5 days, and the fusion drug is automatically targeted to forming thrombi and sites of vascular injury. The fusion drug can thus be used to treat cardiovascular diseases, e.g., as adjunctive therapy to inhibit reocclusions in a patient after thrombolytic therapy or angioplasty.

Compositions and methods for the delivery of drugs by platelets for the

A method of adjunctive therapy to inhibit reocclusions in a patient, e.g., after thrombolytic therapy or angioplasty, by administering to the patient a bolus of an amount of purified pro-urokinase ('pro-UK') that inhibits the formation of occlusive thrombi without inducing a systemic effect in the patient, the pro-UK is administered after the completion of the thrombolytic treatment and periodically thereafter for the duration of therapy, and becomes incorporated into the outer membrane of the platelets of the patient, thereby increasing the T.sub. 1/2 of the pro-UK in plasma, which is about 7 to 8 minutes, to about 4 to 5 days, and inhibiting reocclusion without inducing a systemic effect.

Use of intra-platelet urokinase-type plasminogen activators for

The invention relates to thrombolytically active pro-urokinase (pro-UK) mutants comprising the amino acid sequence of native pro-UK, but including a mutation which causes the pro-UK mutants to induce less fibrinogenolysis and non-specific plasminogen activation than native pro-UK, to have at least a 10-fold lower intrinsic activity than native pro-UK, and to have substantially the same fibrin promotion and thrombolytic activity after plasmin activation compared to native pro-UK when administered to a patient.

Pro-urokinase mutants

The existence of high fibrin-affinity urokinase is discovered by an isolation procedure using fibrin precipitated on an adsorptive-solid matrix. By the method described, the high affinity form of plasminogen activator can be isolated directly from urine or from kidney tissue culture medium. The method is economical and provides a relatively high yield of the activator. The high affinity that this plasminogen activator has for fibrin is a property that makes it an improved thrombolytic agent and when radiolabelled provides a new diagnostic agent for the specific detection of fibrin thrombi through nuclear scanning. The newly-isolated plasminogen activator has the following characteristics: a molecular weight of about 56,000 Daltons, a specific activity of about 40,000-50,000 CTA units/mg, the appearance of a single chain structure and a high affinity for fibrin.

Growing community of inventors

Cambridge, MA, United States of America

Victor Gurewich