30 results about "Cross linked hemoglobin" patented technology

An organ perfusion apparatus and method monitor, sustain and / or restore viability of organs and preserve organs for storage and / or transport. The method includes perfusing the organ at hypothermic and / or normothermic temperatures, preferably after hypothermic organ flushing for organ transport and / or storage. The method can be practiced with prior or subsequent static or perfusion hypothermic exposure of the organ. Organ viability is restored by restoring high energy nucleotide (e.g., ATP) levels by perfusing the organ with a medical fluid, such as an oxygenated cross-linked hemoglobin-based bicarbonate medical fluid, at normothermic temperatures. In perfusion, organ perfusion pressure is preferably controlled in response to a sensor disposed in an end of tubing placed in the organ, by a pneumatically pressurized medical fluid reservoir, providing perfusion pressure fine tuning, overpressurization prevention and emergency flow cut-off. In the hypothermic mode, the organ is perfused with a medical fluid, preferably a simple crystalloid solution containing antioxidants, intermittently or in slow continuous flow. The medical fluid may be fed into the organ from an intermediary tank having a low pressure head to avoid organ overpressurization. Preventing overpressurization prevents or reduces damage to vascular endothelial lining and to organ tissue in general. Viability of the organ may be automatically monitored, preferably by monitoring characteristics of the medical fluid perfusate. The perfusion process can be automatically controlled using a control program.

An organ perfusion apparatus and method monitor, sustain and / or restore viability of organs and preserve organs for storage and / or transport. Other apparatus include an organ transporter, an organ cassette and an organ diagnostic device. The method includes perfusing the organ at hypothermic and / or normothermic temperatures, preferably after hypothermic organ flushing for organ transport and / or storage. The method can be practiced with prior or subsequent static or perfusion hypothermic exposure of the organ. Organ viability is restored by restoring high energy nucleotide (e.g., ATP) levels by perfusing the organ with a medical fluid, such as an oxygenated cross-linked hemoglobin-based bicarbonate medical fluid, at normothermic temperatures. In perfusion, organ perfusion pressure is preferably controlled in response to a sensor disposed in an end of tubing placed in the organ, by a pneumatically pressurized medical fluid reservoir, providing perfusion pressure fine tuning, overpressurization prevention and emergency flow cut-off. In the hypothermic mode, the organ is perfused with a medical fluid, preferably a simple crystalloid solution containing antioxidants, intermittently or in slow continuous flow. The medical fluid may be fed into the organ from an intermediary tank having a low pressure head to avoid organ overpressurization. Preventing overpressurization prevents or reduces damage to vascular endothelial lining and to organ tissue in general. Viability of the organ may be automatically monitored, preferably by monitoring characteristics of the medical fluid perfusate. The perfusion process can be automatically controlled using a control program.

An organ perfusion apparatus and method monitor, sustain and / or restore viability of organs and preserve organs for storage and / or transport. Other apparatus include an organ transporter, an organ cassette and an organ diagnostic device. The method includes perfusing the organ at hypothermic and / or normothermic temperatures, preferably after hypothermic organ flushing for organ transport and / or storage. The method can be practiced with prior or subsequent static or perfusion hypothermic exposure of the organ. Organ viability is restored by restoring high energy nucleotide (e.g., ATP) levels by perfusing the organ with a medical fluid, such as an oxygenated cross-linked hemoglobin-based bicarbonate medical fluid, at normothermic temperatures. During the period in which the organ is preserved and / or maintained, various drug research and development may be performed on and / or with the organ. The organ may be perfused with a fluid containing a substance such as a test substance to obtain data regarding the organ, the substance and an interaction of the substance and the organ. The data may then be used to ultimately provide information regarding the drugs efficacy in support of regulatory filings for new drugs.

An organ perfusion apparatus and method monitor, sustain and / or restore viability of organs and preserve organs for storage and / or transport. Other apparatus include an organ transporter, an organ cassette and an organ diagnostic device. The method includes perfusing the organ at hypothermic and / or normothermic temperatures, preferably after hypothermic organ flushing for organ transport and / or storage. The method can be practiced with prior or subsequent static or perfusion hypothermic exposure of the organ. Organ viability is restored by restoring high energy nucleotide (e.g., ATP) levels by perfusing the organ with a medical fluid, such as an oxygenated cross-linked hemoglobin-based bicarbonate medical fluid, at normothermic temperatures. In perfusion, organ perfusion pressure is preferably controlled in response to a sensor disposed in an end of tubing placed in the organ, by a pneumatically pressurized medical fluid reservoir, providing perfusion pressure fine tuning, overpressurization prevention and emergency flow cut-off. In the hypothermic mode, the organ is perfused with a medical fluid, preferably a simple crystalloid solution containing antioxidants, intermittently or in slow continuous flow. The medical fluid may be fed into the organ from an intermediary tank having a low pressure head to avoid organ overpressurization. Preventing overpressurization prevents or reduces damage to vascular endothelial lining and to organ tissue in general. Viability of the organ may be automatically monitored, preferably by monitoring characteristics of the medical fluid perfusate. The perfusion process can be automatically controlled using a control program.

The present invention includes compositions containing carboxamidomethylated cross-linked hemoglobin where the cysteine moiety of the hemoglobin includes a thiol protecting group and where the hemoglobin has a reduced ability to bind with nitric oxide. Preferably, the hemoglobin is deoxygenated, endotoxin free, and stroma free. The present invention also includes method of preparation, process of preparation and the method of use including supplementing blood volume in mammals and treating disorders in mammals where oxygen delivery agents are of benefit.

Methods for preparation of a heat stable hemoglobin based oxygen-carrier-containing pharmaceutical composition such that beta-beta cross-linking is favored are provided. Using the methods of the present invention, the oxygen affinity of the resultant molecule can be controlled so that hemoglobin based oxygen carriers tailored for specific applications can be produced. Lower oxygen affinity crosslinked hemoglobin is useful for applications requiring rapid tissue oxygenation (e.g. hemorrhagic shock) while higher oxygen affinity cross-linked hemoglobin is useful for applications requiring a slower rate of oxygenation (e.g. cancer adjunct therapy). A highly purified and heat stable crosslinked non-polymeric tetrameric hemoglobin having beta-beta cross-linking of greater than 40-60% and suitable for use in mammals without causing renal injury and vasoconstriction is produced. A high temperature and short time (HTST) heat processing step is performed to effectively remove any undesired dimeric hemoglobin, non-crosslinked tetrameric hemoglobin, and plasma protein impurities.

An organ perfusion apparatus and method monitor, sustain and / or restore viability of organs and preserve organs for storage and / or transport. Other apparatus include an organ transporter, an organ cassette and an organ diagnostic device. The method includes perfusing the organ at hypothermic and / or normothermic temperatures, preferably after hypothermic organ flushing for organ transport and / or storage. The method can be practiced with prior or subsequent static or perfusion hypothermic exposure of the organ. Organ viability is restored by restoring high energy nucleotide (e.g., ATP) levels by perfusing the organ with a medical fluid, such as an oxygenated cross-linked hemoglobin-based bicarbonate medical fluid, at normothermic temperatures. In perfusion, organ perfusion pressure is preferably controlled in response to a sensor disposed in an end of tubing placed in the organ, by a pneumatically pressurized medical fluid reservoir, providing perfusion pressure fine tuning, overpressurization prevention and emergency flow cut-off. In the hypothermic mode, the organ is perfused with a medical fluid, preferably a simple crystalloid solution containing antioxidants, intermittently or in slow continuous flow. The medical fluid may be fed into the organ from an intermediary tank having a low pressure head to avoid organ overpressurization. Preventing overpressurization prevents or reduces damage to vascular endothelial lining and to organ tissue in general. Viability of the organ may be automatically monitored, preferably by monitoring characteristics of the medical fluid perfusate. The perfusion process can be automatically controlled using a control program.

The present invention includes compositions containing carboxymethylated cross-linked hemoglobin where the cysteine moiety of the hemoglobin includes thiol protecting group and where the hemoglobin is incapable of binding with the nitric oxide. Preferably, the hemoglobin is deoxygenated, endotoxin free, and stroma free. The present invention also includes method of preparation, process of preparation and the method of supplementing blood in mammals.

The present invention includes compositions containing carboxymethylated cross-linked hemoglobin where the cysteine moiety of the hemoglobin includes thiol protecting group and where the hemoglobin is incapable of binding with the nitric oxide. Preferably, the hemoglobin is deoxygenated, endotoxin free, and stroma free. The present invention also includes method of preparation, process of preparation and the method of supplementing blood in mammals.

The invention discloses a cross-linked hemoglobin preparation method, wherein the preparation method comprises the following steps: a, diluting deoxidized hemoglobin, and adding the diluted deoxidizedhemoglobin into an anaerobic reactor; b, arranging an atomizer at the top of the anaerobic reactor, adding a cross-linking agent into the reactor through the atomizer, and carrying out a cross-linking reaction; and c, adding a reducing agent to terminate the cross-linking reaction, and removing the unreacted cross-linking agent and the terminating reducing agent in an ultrafiltration liquid changing manner to obtain the cross-linked hemoglobin. The cross-linked hemoglobin prepared by the preparation method contains low contents of macromolecules (greater than 500 kD) and micromolecules (lessthan 64 kD); by adding the cross-linked hemoglobin into organ perfusate, the phenomena of organ acidosis and lysosomal enzyme activation adverse consequences caused by lack of an oxygen carrying agentin conventional perfusate and cell damage caused by excessive free radicals generated during reperfusion can be solved, and the perfusate is quite simple and convenient to prepare, store, transport and use.

A process for preparing the cross-linked haematoglobin used as the substitute of red blood cell includes such steps as deoxygenating haematoglobin solution, regulating pH=6.0-7.5, mixing with polyamino compound as crosslinking competition inhibitor, dispersing glutaraldehyde in it, reaction, adding the said competition inhibitor, reduction reaction, and filtering and ultra-filtering.

The present invention relates to conjugates of hemoglobin and human serum albumin and the preparation process thereof. The conjugates have molecular weight in the range of 100-300 kD, comprising 1-3 intermolecularly or intramolecularly cross-linked hemoglobin molecules and 1-3 human serum albumin molecules. The conjugates are obtained by the following steps: preparing stroma-free hemoglobin, then coupling hemoglobin to human serum albumin and purifying the products using anion exchange chromatography. The hemoglobin and human serum albumin conjugates have characteristics suitable for being used as blood substitutes.

A method heat treatment of cross-linked hemoglobin solutions including polymeric hemoglobin is disclosed. The method involves contacting the hemoglobin solution with a high temperature short time heat treatment apparatus. The high temperature short time process thermally denatures unmodified tetrameric hemoglobin (hemoglobin dimer form), protein impurities (e.g. immunoglobin-G, serum albumin), bacteria, and viruses so that renal injury, vascular detrimental effects and other toxicity reactions can be avoided.

Methods for preparation of a heat stable hemoglobin based oxygen-carrier-containing pharmaceutical composition such that beta-beta cross-linking is favored are provided. Using the methods of the present invention, the oxygen affinity of the resultant molecule can be controlled so that hemoglobin based oxygen carriers tailored for specific applications can be produced. Lower oxygen affinity crosslinked hemoglobin is useful for applications requiring rapid tissue oxygenation (e.g. hemorrhagic shock) while higher oxygen affinity cross-linked hemoglobin is useful for applications requiring a slower rate of oxygenation (e.g. cancer adjunct therapy). A highly purified and heat stable crosslinked non-polymeric tetrameric hemoglobin having beta-beta cross-linking of greater than 40-60% and suitable for use in mammals without causing renal injury and vasoconstriction is produced. A high temperature and short time (HTST) heat processing step is performed to effectively remove any undesired dimeric hemoglobin, non-crosslinked tetrameric hemoglobin, and plasma protein impurities.

A process for efficiently preparing bis-tetrameric hemoglobin in which the tetramers are specifically linked at predetermined sites on the β-sub-units and in which the tetramers themselves are effectively bonded to prevent dissociation into dimeric αβ-hemoglobin sub-units therefrom is provided. The process uses as cross-linking reagent a hexafunctional aromatic acyl phosphate containing amide groups, The concept is to use a cross-linking reagent which has an excess of site-specific hemoglobin reacting groups, namely six acyl phosphate groups, so that at least four of them will react site specifically to form the bis-tetrameric product of particular therapeutic interest.

The present invention includes compositions containing carboxamidomethylated cross-linked hemoglobin where the cysteine moiety of the hemoglobin includes a thiol protecting group and where the hemoglobin has a reduced ability to bind with nitric oxide. Preferably, the hemoglobin is deoxygenated, endotoxin free, and stroma free. The present invention also includes method of preparation, process of preparation and the method of use including supplementing blood volume in mammals and treating disorders in mammals where oxygen delivery agents are of benefit.

The invention relates to a preparation method of polymerized hemochrome low in content of polymer oligomers, and particularly provides a preparation method of cross-linking hemochrome. The preparationmethod comprises the steps of a, diluting deoxyhemoglobin; b, enabling a cross-linking agent namely glutaraldehyde and the diluted deoxyhemoglobin to be subjected to a cross-linking reaction; c, adding a reducing agent to end the cross-linking reaction, removing the unreacted cross-linking agent and the reducing agent for ending the cross-linking reaction in the manner of an ultrafiltration liquid exchange manner so as to obtain the cross-linking hemochrome, wherein in the step a, the concentration of the diluted deoxyhemoglobin achieves 1-3g / dL, and N- acetyl-L- cysteine is added to the hemochrome, so that the mass ratio of the hemochrome to the N- acetyl-L- cysteine in the solution is (5-10) to 1, and the mass ratio of the hemochrome to the cross-linking agent is 1000 to (29-45 ). The polymerized hemochrome prepared by the method is low in content of dimers and low in content of polymer oligomers.

A method for separating unmodified hemoglobin from cross-linked hemoglobin in a hemoglobin solution. The method involves contacting the hemoglobin solution with a least one dissociating agent to form a dissociation solution wherein unmodified tetrameric hemoglobin is dissociated to form hemoglobin dimers. The hemoglobin dimers are then separated from the dissociation solution, while retaining the cross-linked hemoglobin in the dissociation solution.

The present invention relates to nanometer material coated internally cross-linked hematoglobin nanocapsule and its preparation process. The pyridoxal phosphate internally cross-linked hematoglobin nanocapsule includes nanometer chitosan and sodium alginate as coating material and pyridoxal phosphate internally cross-linked hematoglobin, has diameter of 60-120 nm, and possesses excellent oxygen carrying capacity, excellent biocompatibility and no adverse effect on various blood components. The preparation process uses no organic solvent and easy-to-decompose nanometer coating, resulting in no toxicity. The preparation process is simple, fast and expected for large scale application.

The invention discloses a dextran cross-linked hemoglobin-based oxygen carrier, a preparation method thereof and an application. The preparation method includes the steps: oxidizing hydroxy on dextran 40 into an aldehyde group by sodium periodate to obtain activated dextran; protecting sulfydryl on hemoglobin by 4-PDS; crosslinking the activated dextran with the bovine hemoglobin closed by the sulfydryl; finally, de-protecting the sulfydryl to obtain the dextran cross-linked hemoglobin-based oxygen carrier. P50 of the dextran cross-linked hemoglobin-based oxygen carrier (Dex-Hb) prepared by the method can be effectively increased, oxygen carrying-releasing capacity is improved, intramolecular crosslinking of hemoglobin tetramer is realized, so that tetramer stability of HBOCs is remarkably enhanced, and physiological functions of the HBOCs are further improved.

The present invention relates to novel methods of using blood substitutes to treat acute blood loss and novel pharmaceutical compositions comprising blood substitutes. Blood substitutes useful for the methods of the present invention can (1) induce expression of erythropoietin as tested in a cell culture under normoxic conditions, and / or (2) induce erythropoiesis under normoxic conditions as measured by (a) a decrease in the doubling time of the subject's hematocrit or hemoglobin, and / or (b) an increase in the subject's circulating erythropoietin level. Blood substitutes useful for the pharmaceutical compositions of the present invention can (1) stabilize HIF-1 alpha expression, and / or (2) down regulate NF-kappa B. Preferably, the blood substitutes are cross-linked hemoglobin blood substitutes, or more preferably, cross-linked hemoglobins that comprise a hemoglobin that is cross-linked intramolecularly with periodate-oxidized ATP, cross-linked intermolecularly with periodate-oxidized adenosine, and conugated with reduced glutathione.

A method for preparing red blood cell substitutes with polyaldehyde-based compounds. In the reaction of polymerizing hemoglobin, the present invention reduces the concentration of the reaction substrate, and quickly adds the polyaldehyde-based compound solution to the hemoglobin solution for reaction; after the reaction is terminated, the obtained molecular weight is 100 The hemoglobin polymer between ~1500kDa; Compared with the prior art, the present invention has: 1) fast addition of polyaldehyde compounds, which greatly shortens the polymerization process, can shorten the production cycle and reduce costs; The compound maintains a high concentration for a relatively long period of time and has the function of inactivating the virus; ③More than 90% of the hemoglobin molecules are cross-linked within the molecule, making the structure of hemoglobin stable; ④Higher pH value can improve the reaction solution. The concentration of free polyaldehyde compounds in the medium can reduce the content of uncrosslinked hemoglobin between molecules, which can reduce toxicity; ⑤The average molecular weight of hemoglobin after polymerization increases significantly, and its half-life can reach 18-25h, which is more conducive to clinical application.

The invention relates to a method for preparing polymerized hemoglobin with low polymer content, and specifically provides a method for preparing cross-linked hemoglobin, including: a. diluting deoxygenated hemoglobin; b. cross-linking agent glutaraldehyde and the diluted deoxygenated hemoglobin Linking reaction; c, add reducing agent to terminate crosslinking reaction, remove unreacted crosslinking agent and stop reducing agent with the mode of ultrafiltration changing liquid, obtain crosslinked hemoglobin; Wherein: in described step a, dilute deoxygenated hemoglobin concentration to 1-3g / dL, add N-acetyl-L-cysteine ​​in described hemoglobin, make the mass ratio of hemoglobin and N-acetyl-L-cysteine ​​in the solution be 5-10:1; And hemoglobin The mass ratio to the crosslinking agent is 1000:29‑45. In the polymerized hemoglobin prepared by the method, not only the dimer content is low, but also the high polymer content is also low.

The invention discloses a dextran cross-linked hemoglobin-based oxygen carrier, a preparation method thereof and an application. The preparation method includes the steps: oxidizing hydroxy on dextran 40 into an aldehyde group by sodium periodate to obtain activated dextran; protecting sulfydryl on hemoglobin by 4-PDS; crosslinking the activated dextran with the bovine hemoglobin closed by the sulfydryl; finally, de-protecting the sulfydryl to obtain the dextran cross-linked hemoglobin-based oxygen carrier. P50 of the dextran cross-linked hemoglobin-based oxygen carrier (Dex-Hb) prepared by the method can be effectively increased, oxygen carrying-releasing capacity is improved, intramolecular crosslinking of hemoglobin tetramer is realized, so that tetramer stability of HBOCs is remarkably enhanced, and physiological functions of the HBOCs are further improved.

The disclosure concerns a class of hemoglobin based oxygen carriers (HBOCs) comprising a first hemoglobin protein cross-linked by a chemical reaction that is followed by a strain-promoted alkyne-azide cycloaddition (SPAAC) reaction or by a strain-promoted alkyne-nitrone cycloaddition (SPANC) in the absence of added copper salts to a second modified cross-linked hemoglobin protein. The resulting construct is an HBOC that is capable of binding oxygen and releasing same in a useful manner upon addition in an appropriate solution to the circulatory system of a patient. The disclosure also concerns a method of production of the HBOC where a first and second hemoglobin protein are produced by covalently linking hemoglobin to an angle strained cycloalkyne moiety. A compound comprising at least 2 azide or nitrone moieties is then added for reacting under conditions conducive to SPAAC or SPANC in reaction in absence of copper ions with the first and second hemoglobin protein for causing covalent linkage of the first and second hemoglobin protein to the compound comprising said at least 2 azide or nitrone moieties, resulting in said HBOC. The HBOC can be used for transfusion, perfusion or for increasing oxygen transport.

The present invention relates to nanometer material coated internally cross-linked hematoglobin nanocapsule and its preparation process. The pyridoxal phosphate internally cross-linked hematoglobin nanocapsule includes nanometer chitosan and sodium alginate as coating material and pyridoxal phosphate internally cross-linked hematoglobin, has diameter of 60-120 nm, and possesses excellent oxygen carrying capacity, excellent biocompatibility and no adverse effect on various blood components. The preparation process uses no organic solvent and easy-to-decompose nanometer coating, resulting in no toxicity. The preparation process is simple, fast and expected for large scale application.