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Methods and compositions for safe and effective thrombolysis

A technology of composition and use, applied in the field of safe and effective thrombolysis, which can solve the problems of delayed treatment and the like

Active Publication Date: 2021-07-02
THROMBOLYTIC SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, correctly diagnosing the specific cause of a stroke requires a CT scan or MRI, which if not immediately available may delay treatment
In the absence of such a proper diagnosis, if the cause happens to be bleeding rather than a blood clot, administration of clot-dissolving agents such as tissue plasminogen activator ("tPA") can be very dangerous, as administered to patients with intracerebral hemorrhage tPA or other thrombolytics can be fatal

Method used

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  • Methods and compositions for safe and effective thrombolysis
  • Methods and compositions for safe and effective thrombolysis
  • Methods and compositions for safe and effective thrombolysis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0095] Example 1. Maximum clot lysis rate of in vitro thrombolysis induced by tPA / M5

[0096] The fibrinolytic and fibrinolytic effects of specific combinations of tPA and M5 were studied in the context of human plasma in vitro.

[0097] Material

[0098]mproUK(M5) comprising the replacement of lysine at amino acid position 300 of prourokinase with histidine (Lys300→His) was prepared from E. coli by the company PxTherapeutics (Grenoble, France). Tissue plasminogen activator (tPA) was obtained from Genentech (South San Francisco, CA). Human fibrinogen (Kabi, grade L) was obtained from Chromogenix, Milan, Italy. Aprotinin and fluorescein isothiocyanate were obtained from Sigma Chemicals (St. Louis, MO). Thrombin (ThromboMax, 100 NIH U / ml) was obtained from Sigma (St Louis, MO). Clinical grade C1-inhibitors were obtained from CSL Behring, Marburg, Germany. Pooled human expired blood bank plasma from four donors was used in the experiments.

[0099] Note that various anima...

Embodiment 2

[0126] Example 2. In vivo thrombolysis by combination of tPA and M5

[0127] Anesthetize a male mutt weighing 10-15 kg with sodium pentobarbital and maintain breathing chamber air. Blood clots were formed from 1 ml of native whole canine blood (as described in US Patent No. 7,074,401), to which radiolabeled fibrinogen (1.9 μCi, 0.75 mCi / mg protein) and thrombin (10 units) were added. After 20 min, the clot was washed 3 times with brine, and then cut into small pieces (about 1mm 3 ) and injected into the femoral vein through a 16G needle. After 15 min, a blood sample was obtained from a cannula in the contralateral femoral vein for measurement of baseline radioactivity.

[0128] Dogs were divided into four groups and injected with: (1) saline, (2) 2-5 mg tPA bolus, (3) intravenous infusion of M5 (20 μg / kg / min) for 60 min, or (4) 2-5 mg tPA bolus dose, followed by intravenous infusion of M5 (20 μg / kg / min) for 60 minutes. Blood samples were obtained at intervals during the in...

Embodiment 3

[0129] Example 3. Characterization of C1-inhibitors and M5 in a rat model of intracerebral hemorrhage (ICH)

[0130] The aim of this study was to investigate the effect of M5 on the volume of intracerebral hemorrhage (ICH).

[0131] Twenty adult male Sprague-Dawley rats were used for the study. Rats were randomly selected for the day of surgery. Rats were given a unique identification number by tail tag. Animals received cefazolin sodium intraperitoneally (40 mg / kg; Hospira 101C049) and buprenorphine subcutaneously (1 mg / kg; Reckitt Benckiser, 219202) immediately before the start of surgery. While rats under isoflurane anesthesia (1.5%-2%) breathed spontaneously inside a nitrous oxide / oxygen mixture (2:1), a small burr hole was drilled and 10 μL of 30G was microinjected into A needle (Hamilton, 700 series) was slowly lowered into the right striatum at the following coordinates from bregma: 0.0 mm anteriorly, 3 mm laterally and 6 mm deep. Over a period of 3 min, 3 μL of sal...

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Abstract

Provided herein are methods, compositions and kits for safe and effective thrombolysis, eg, for the treatment of underlying stroke or acute myocardial infarction (AMI).

Description

technical field [0001] The present invention relates to methods and compositions for safe and effective thrombolysis. [0002] Background of the invention [0003] Thrombosis occurs when a blood clot (thrombus) forms inside a blood vessel and blocks blood flow through the circulatory system. When a blood vessel is damaged, the body uses platelets and fibrin to form a blood clot to seal the damaged vessel and prevent blood loss. This process is called hemostasis. Even when blood vessels are not damaged, blood clots can form in the body under certain conditions. Blood clots consist primarily of aggregated platelets and a network of cross-linked fibrin, the natural polymer of fibrinogen in blood. When the thrombus is large enough to reduce blood flow to the tissue, hypoxia / anoxia can occur, leading to tissue damage and even tissue death. Depending on where it is in the arterial system - heart, brain or leg, a blood clot can trigger a heart attack, stroke or peripheral gangre...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K38/49A61K38/57A61P7/02A61P9/10
CPCA61K9/0019A61K38/49A61K38/57A61P9/10C12Y304/21068C12Y304/21073A61K2300/00
Inventor 维克多·古里维希
Owner THROMBOLYTIC SCI
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