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A heat stable oxygen carrier-containing pharmaceutical composition for different treatment applications

A composition and heat-stabilized technology, applied in the direction of drug combination, antineoplastic drugs, pharmaceutical formulations, etc., can solve the problems of unacceptable high percentage of dimer units, hemoglobin composition cannot be satisfactorily administered to mammals, etc.

Active Publication Date: 2014-03-26
BILLION KING INT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] Further problems with prior art attempts to stabilize hemoglobin include: tetrameric hemoglobin produced contains an unacceptably high percentage of dimer units; the presence of dimers makes the hemoglobin composition unsatisfactory administered to mammals
[0011] Further problems with prior art attempts to form stable hemoglobin include the presence of protein impurities such as immunoglobulin G, which can cause allergic effects in mammals

Method used

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  • A heat stable oxygen carrier-containing pharmaceutical composition for different treatment applications
  • A heat stable oxygen carrier-containing pharmaceutical composition for different treatment applications
  • A heat stable oxygen carrier-containing pharmaceutical composition for different treatment applications

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0081] Process overview

[0082] A schematic flow diagram of the method of the present invention is shown in figure 1middle. Bovine whole blood was collected in closed sterile containers / bags containing 3.8% (w / v) trisodium citrate solution as anticoagulant. The blood is then mixed immediately and thoroughly with trisodium citrate solution to inhibit blood clotting. Red blood cells (RBCs) are separated from plasma and other smaller blood cells and collected by an apheresis mechanism. For this step, a "cell washer" was used with gamma sterilized disposable centrifuge bowls. RBCs were washed with an equal volume of 0.9% (w / v NaCl) saline.

[0083] Washed red blood cells are lysed to release the hemoglobin contents by subjecting the red blood cell membrane to a hypotonic shock. figure 2 The dedicated point-of-care cell lysis device for the RBC lysis device drawn in was used for this purpose. After RBC lysis, hemoglobin molecules were separated from other proteins by tangen...

Embodiment 2

[0086] Timed and controlled hypotonic lysis and filtration

[0087] Fresh bovine whole blood was collected and shipped under chilled conditions (2-10°C). Red blood cells were separated from plasma by a cell washer, followed by 0.65 μm filtration of red blood cells. After washing the red blood cell (RBC) filtrate with 0.9% saline, the filtrate was disrupted by hypotonic lysis. by using figure 2 The instant cell lysis device drawn in is used for hypotonic lysis. The instant cell lysis device includes a static mixer to aid in cell lysis. An RBC suspension containing controlled hemoglobin concentration (12-14 g / dL) was mixed with 4 volumes of purified water to generate a hypotonic shock to the RBC cell membranes. The period of hypotonic shock is controlled to avoid undesired lysis of leukocytes and platelets. The hypotonic solution is passed through the static mixer section of the instant cell lysis device for 2-30 seconds, or otherwise sufficient to lyse red blood cells, pr...

Embodiment 3

[0090] Virus clearance studies on stroma-free hemoglobin solutions

[0091] In order to demonstrate the safety of the product described in the present invention, we demonstrated the virus clearance capacity of (1) 0.65 μm diafiltration step and (2) 100 kDa ultrafiltration step through virus validation studies. This was done by intentionally adding different model viruses (encephalomyocarditis virus, pseudorabies virus, bovine viral diarrhea virus and bovine parvovirus) to both processes in scale-down form. (bovine parvovims)). Four types of viruses were used in this study (see Table 3). These viruses differ in their biophysical and structural characteristics and exhibit variations in resistance to physical and chemical agents or treatments.

[0092] table 3

[0093]

[0094] The verification scheme is briefly shown in Table 4 below.

[0095] Table 4

[0096]

[0097] A summary of the results for the logarithmic (log) reduction of the four viruses in (1) 0.65 μm diaf...

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Abstract

A highly purified and heat stable cross-linked nonpolymeric tetrameric hemoglobin suitable for use in mammals without causing renal injury and vasoconstriction is provided. A high temperature and short time (HTST) heat processing step is performed to remove undesired dimeric form of hemoglobin, uncross-linked tetrameric hemoglobin, and plasma protein impurities effectively. Addition of N-acetyl cysteine after heat treatment and optionally before heat treatment maintains a low level of met-hemoglobin. The heat stable cross-linked tetrameric hemoglobin can improve and prolong oxygenation in normal and hypoxic tissue. In another aspect, the product is used in the treatment of various types of cancer such as leukemia, colorectal cancer, lung cancer, breast cancer, liver cancer, nasopharyngeal carcinoma and esophageal cancer.The inventive tetrameric hemoglobin can also be used to prevent tumor metastasis and recurrence following surgical tumor excision. Further the inventive tetrameric hemoglobin can be administered to patients prior to chemotherapy and radiation treatment.

Description

[0001] Cross References to Related Applications [0002] This application is an International Patent Application claiming priority to US Continuation-in-Part No. 13 / 179,590, currently Patent No. 8,048,856, filed July 11, 2011, the disclosure of which is hereby incorporated by reference. [0003] Copyright Notice / Permission [0004] Portions of the disclosure of this patent document contain material that is protected by copyright. The copyright owner has no objection to the facsimile printing by anyone of the patent document or the patent disclosure as it appears in the USPTO patent files or records, but otherwise fully reserves all copyright rights. The following notice applies to the methods, experiments and data described below and in the accompanying figures: Copyright 2010, Billion King International Limited, all rights reserved. technical field [0005] The present invention relates to a method for the preparation of a pharmaceutical composition containing a thermosta...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K38/42A61K47/48A61K47/30A61P35/00
CPCA61K38/42A61P35/00A61K47/50A61K47/30
Inventor 黄炳镠郭瑞仪刘思行
Owner BILLION KING INT
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