WO1997018000A1 - S-nitroso-hemoglobine et ses utilisations therapeutiques - Google Patents

S-nitroso-hemoglobine et ses utilisations therapeutiques Download PDF

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Publication number
WO1997018000A1
WO1997018000A1 PCT/US1996/017508 US9617508W WO9718000A1 WO 1997018000 A1 WO1997018000 A1 WO 1997018000A1 US 9617508 W US9617508 W US 9617508W WO 9718000 A1 WO9718000 A1 WO 9718000A1
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hemoglobin
human hemoglobin
nitroso
nitrosylated
oxygen
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PCT/US1996/017508
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English (en)
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Jonathan Stamler
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Brigham And Women's Hospital
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Priority to AU75297/96A priority Critical patent/AU7529796A/en
Publication of WO1997018000A1 publication Critical patent/WO1997018000A1/fr

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/795—Porphyrin- or corrin-ring-containing peptides
    • C07K14/805—Haemoglobins; Myoglobins
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
    • A61M2202/00—Special media to be introduced, removed or treated
    • A61M2202/02—Gases
    • A61M2202/0266—Nitrogen (N)
    • A61M2202/0275—Nitric oxide [NO]

Definitions

  • Hemoglobin and other blood substitutes and oxygen delivery systems cause vascular constriction in the entire cardiovascular system, particularly in the lung and gastrointestinal tract by causing the contraction of vascular smooth muscle. Further, hemoglobins and other oxygen delivery systems that consume or scavenge nitric oxide thereby cause vasoconstriction which hinders oxygen delivery and thereby raises blood pressure. Overcoming these limitations would end a lengthy quest for an alternative to red blood cell transfusions and satisfy a strong medical need.
  • Cell-free blood substitute compositions sought to be used for treating hemorrhage and preserving organs and tissue for transplantation have predominantly been preparations of heme-containing proteins.
  • Numerous investigators and foreign and domestic companies have sought to produce isolated human, bovine and other hemoglobin and modified hemoglobins for use in cell-free blood substitute preparations. Native hemoglobin has been isolated from blood and used in such preparations.
  • Modified hemoglobins have been prepared and used for such cell-free preparations.
  • cross-linked polymerized hemoglobins see U.S. Patent No. 5, 194,590
  • cross-linked polymerized pyridoxylated hemoglobin U.S. Patent No. 5, 194,590 and 4,826,81 1
  • Multimeric hemoglobin-like proteins based on pseudo-tetramer containing pseudo-dimer polypeptides with globin-like domains have been used in order to prolong hemoglobin half life (see WO 93/09143).
  • Variants such as conjugates of hemoglobin with other drugs are also used (see WO 93/08842).
  • Di-alpha-globin-like polypeptide and di-beta-glob -like polypeptides connected into a single chain and inco ⁇ orating heme have been used to prepare a human hemoglobin-like protein (see WO 90/13645).
  • Hemoglobin mutants having lower O 2 -affin ⁇ ty have been prepared by recombinant techniques and used as blood substitutes (see WO 88/09179).
  • Hemoglobin and heme-containing protem based blood substitute compositions are disclosed, inter alia, in U.S. Patent Nos.5,217,648; 5, 194,590; 5,061 ,688; 4,826,811;5,281,579; 5, 128,452; 5,248,766; 5,041 ,615; 4,861 ,867; 4,831 ,012; 5,296,465; 5,084,558; 5,295,944; 4,780,210; 4,925,574;5,264,555 and numerous others; and in PCT publication nos.
  • endothelium- de ⁇ ved relaxing factor a vascular relaxing factor, known as endothelium- de ⁇ ved relaxing factor (EDRF), which has been identified as nitric oxide (NO), or a closely related derivative thereof.
  • EDRF endothelium- de ⁇ ved relaxing factor
  • NO nitric oxide
  • S-nitrosothiols are adducts that form readily under physiologic conditions from the reaction of NO with reduced low molecular weight thiols (Oae et al., Org. Prep. Proc. Int. , 15(3): 165-198, 1983). These compounds have half-lives that are significantly greater than that of NO and, like EDRF, possess vasorelaxant activity that is mediated, in part, through activation of guanylate cyclase (Kowaluk et al., J. Pharmacol. Exp. Ther., 256: 1256-1264, 1990; Loscalzo et al., J. Pharmacol. Exp. Ther. 249(3):126-129, 1989; and Ignarro et al., J. Pharmacol. Exp. Ther., 2I8(3):139- 749, 1981).
  • Hemoglobin is a globular protein, which binds reversibly to blood oxygen through passive diffusion from entry of air into the lungs. Hemoglobin- oxygen binding greatly increases the capacity of the blood to transport oxygen to bodily tissues; thus, the binding affinity between hemoglobin and oxygen is a critical factor in determining the level of oxygen transport to the tissues.
  • the thiol group on the hemoglobin molecule regulates the affinity of hemoglobin for oxygen.
  • S-nitrosothiols such as certain S-mtroso-proteins and certain low molecular weight S-nitrosothiols do not react with the iron-binding site of hemoglobin, as does NO, but instead, bind to the thiol group. Thus, methemoglobin formation is prevented and hemoglobin-oxygen binding is unimpaired.
  • Another embodiment of the invention involves the administration of S-NO-hemoglobin or the in vivo nitrosylation of hemoglobin by coadministration of a suitable S-nitrosothiol, to increase the oxygen-carrying capacity of the blood, and oxygen transport by hemoglobin to bodily tissues.
  • these compounds may be useful in the treatment of disorders which are associated with insufficient oxygen transport, or in clinical situations in which increased oxygen transport is needed.
  • Examples of such clinical situations include, but are not limited to, hypoxic disorders resulting from pneumothorax, airway obstruction, paralysis or weakness of the respiratory muscles, inhibition of respiratory centers by drug or other agents, or other instances of decreased pulmonary ventilation, including acute respiratory distress syndrome. Additional clinical indicates include impaired alveolar gas diffusion such as occurs in interstitial fibrosis, bronchiole constriction, pulmonary edema, pneumonia, hemorrhage, drowning, anemias, arteriovenous shunts, and carbon monoxide poisoning.
  • nitroso-hemoglobin may also be used to modulate the delivery of carbon monoxide or nitric oxide (bound to hemoglobin) to bodily tissues.
  • any thiol-containing heme proteins may be nitrosylated and used to enhance the oxygen-carrying capacity of the blood.
  • An additional aspect of the invention is the use of blood substitute compositions in accordance with the invention for maintaining and perfusing transplant organ or tissue materials such that they can be maintained for durations necessary to transport them or to culture expand them, particularly when they are largely comprised of progenitor cells.
  • Such perfusion of organ and tissue maintenance compositions are in liquid form and constitute the compounds of the invention in physiologically acceptable carriers.
  • This invention relates to nitrosylation of blood substitutes, particularly including heme proteins such as hemoglobin as a therapeutic modality.
  • the invention also relates to nitroso-protein compounds and their use as a means to selectively regulate specific protein functions, to selectively regulate cellular function, to endow the protein with new smooth muscle relaxant and platelet inhibitory properties and to provide targeted delivery of nitric oxide to specific bodily sites.
  • An additional aspect of the invention is blood substitute compositions comprising nitrosylated proteins and other constituents. These blood substitute compositions are preferably cell free. Principally useful in such compositions are various forms of nitrosylated hemoglobin and modified hemoglobins.
  • nitrosylated blood substitutes have a vasorelaxant activity which is directly opposite to the vasoconstrictive properties of the non-nitrosylation forms.
  • the administration of the nitrosylated forms effects vasodilation both by adding the administered nitric oxide to the recipient but also by not reducing through scavenging, the constitutive levels.
  • the invention relates to nitrosylation of sites such as sulfhydryl (thiol), oxygen, carbon and nitrogen, present on proteins and amino acids, as a means to achieve the above physiological effects.
  • the therapeutic effects may be achieved by the administration of nitrosylated proteins and amino acids as pharmaceutical compositions, or by nitrosylation of proteins and amino acids in vivo through the administration of a nitrosylating agent, such as in the form of a pharmaceutical composition.
  • the invention provides a method for inhibiting the vasoconstrictive and nitric oxide depleting effects of hemoglobin and heme-containing based blood substitute compositions by the concurrent systemic administration of nitric oxide or a compound which donates, releases or transfers nitric oxide.
  • Such administration can, for example, be by inhalation or intravenously, separately or in composition with the blood substitute.
  • the invention provides S-nitroso hemoglobin, preferably human hemoglobin, and particularly human hemoglobin which has been S-nitrosylated at the thiol moiety of a cysteine in one or both of the 0-subunits of the hemoglobin.
  • the invention particularly relates to a S-nitroso-hemoglobin obtained by the nitrosylation of hemoglobin with an S-nitrosothiol.
  • This compound may be used as a blood substitute or as a therapeutic agent to promote vasodilation and platelet inhibition, and to treat or prevent cardiovascular disorders. It does not suffer from the adverse effects of dangerously high blood pressure observed upon the administration of native hemoglobin.
  • Another aspect of the invention provides a method for enhancing the hypertension protective effect of administering S-nitroso-hemoglobin by co-administering a thiol, such as glutathione or N-acetyl cysteine.
  • a thiol such as glutathione or N-acetyl cysteine.
  • the NO groups of the SNO- hemoglobin will be transferred to the low molecular weight thiols which are more rapid and complete in their release and transfer of NO.
  • the presence of these thiols will act as a catalyst for enhancing the hypotensive effect of the presence of NO.
  • the invention provides a composition comprising a human hemoglobin and a S-nitrosothiol. More particularly, the S-nitrosothiol is selected from the group consisting of S-nitrosocysteine, S-nitroso-N-acetylcysteine, S- nitrosohomocysteine, S-nitrosoglutathione and S-nitrosocysteinylglycine.
  • the invention also provides a method for systemic delivery of nitric oxide and for regulating oxygen delivery to bodily sites by administering pharmaceutical compositions containing S-nitroso-hemoglobin.
  • the invention also provides a method for increasing the capacity of hemoglobin to bind oxygen, comprising administering a therapeutically effective amount of an S- nitrosothiol compound to an animal in need thereof.
  • the invention also provides a method for increasing oxygen transport to bodily tissues, comprising administering a therapeutically effective amount of an S-nitrosothiol compound to an animal in need thereof.
  • the invention also provides a method for the treatment or prevention of disorders associated with insufficient oxygen supply to bodily tissues, comprising administering a therapeutically effective amount of an S-nitrosothiol to an animal in need thereof.
  • the invention provides a method of treating ischaemic syndromes, such as can applied as, for example, post-thrombolytic therapies.
  • the S-nitrosylated- hemoglobins of the invention are effective to treat an individual in need of treatment for ischaemic injury. It is believed that they provide this effect by one or more mechanisms including increased oxygen delivery to the ischemic site by causing vasodilation and by scavenging uncharged nitric oxide and superoxide species from the circulation.
  • the invention provided a method for the treatment of shock. Even though the administration of native hemoglobin has been accompanied by dangerous elevations in blood pressure, S-netrosylated-hemoglobin is effective in controllably elevating dangerously low blood pressures which are associated with shock.
  • the degree to which a given dose of S-nitroso-hemoglobin will elevate a pathologically low blood pressure in a shock patient is proportional to the number of nitrosyl substitutions per hemoglobin molecule of the compound administered.
  • One of ordinary skill in the an will be able to ascertain readily a dose and degree of polynitrosylation optimization for a standardized therapeutic preparation.
  • Figure 1 S-nitrosyiation of hemoglobin, as determined by the Saville method.
  • FIG. 2 UV spectrum of hemoglobin incubated with S-nitroso-N-acetylcysteine (SNOAC).
  • Tracing 5 reflects the product of treating the S-nitroso-oxyhemoglobin, for example, from tracing with dithionitrite.
  • Figure 3 Reaction of nitric oxide at the iron-binding site of hemoglobin.
  • FIG 4 UV spectrum of S-NO-hemoglobin (Hb). Spectra show that oxygen binding to the heme site is largely unaffected at S-NO/Hb stoichiometries (0.05 and 0.37) that are lacking in smooth muscle contractile activity (see Figure 2). Higher ratios of S-NO/Hb (1.59) are associated with methemoglobin formation. Curves for oxy- Fe(II)-hemoglobin (dithionite-treated) and methemoglobin ( 3 Fe(CN)-treated) are shown for comparison.
  • FIG. 5 Reversal of oxyhemoglobin (Hb)-induced contraction of aortic rings by S-nitrosylation. Hemoglobin is shown to constrict vessels in a dose-dependent manner (•). S-NO-Hb, with a stoichiometry of 0.1 S-NO ⁇ Hb, entirely prevents the constrictor response ( o ). Increasing the S-NO-Hb ⁇ Hb stoichiometry to 1 and oxidizing the metal converts hemoglobin into a potent vasodilator ( ⁇ ).
  • S-nitrosothiols deliver NO in its most biologically relevant, and non-toxic form. This is critical, because the pharmacological efficacy of NO depends upon the form in which it is delivered.
  • S-nitrosothiols can deliver NO as charged species, nitrosonium (NO + ) or nitroxyl (NO ), as opposed to the uncharged NO radical (NO*). This is important because the charged species behave in a very different manner from NO* with respect to chemical reactivity.
  • NO nitrosonium and nitroxyl do not react with O 2 or O 2 species to produce toxic oxides of nitrogen, and are also resistant to decomposition in the presence of redox metals.
  • NO thiol group
  • regulated means effective control of the activity of a protein or amino acid, in a selective manner so as to cause the protein or amino acid to exert a desired physiological effect.
  • modified means to effectively alter the activity of a protein or amino acid in a selective manner, so as to cause the protein or amino acid to exert a desired physiological effect.
  • enhanced means to alter effectively the activity of a protein or amino acid in a selective manner, so as to cause an increase or improvement in the activity of the protein or amino acid, or endow the protein or amino acid with additional capabilities.
  • mutant refers to any structurally modified protein or polypeptide that retains the same physiological activity of interest, as the parent protein or polypeptide, whether to a greater or lesser extent. Other properties or advantages may be present in such mutant, variant or fragment compound, such as increased half-life or resistance to natural inhibitors of the parent protein. Mutation can refer, for example, to changes in one or more amino acids in the primary sequence. Variants can refer for example, to posttranslational modifications, such as glycosylation, conformational contraint, or the addition of lipid moieties. Fragments can refer to polypeptides that retain some, all or more of the desired physiological property of the parent protein while not having one or more amino acids or sequences thereof present in the parent protein.
  • substantially homologous refers to amino acid sequences of hemoglobins, modified hemoglobins and other heme-containing proteins, means that a particular subject sequence, for example, a mutant sequence, vanes from a reference sequence by one or more substitutions, deletions, or additions, the net effect of which does not result in an adverse functional dissimilarity between reference and subject sequences.
  • sequences having greater than 90 percent homology, equivalent biological activity, and equivalent expression characteristics are considered substantially homologous.
  • truncation of the mature sequence should be disregarded.
  • Sequences having lesser degrees of homology, comparable bioactivity, and equivalent expression characteristics are considered equivalents.
  • Other embodiments include fusion protein products that exhibit similar oxygen carrying activities. Further embodiments include such proteins that are chemically synthesized as well as any proteins or fragments thereof that are substantially homologous.
  • activity refers to any action exerted by the protein or amino acid which results in a physiological effect.
  • compositions utilized in this invention can be administered by intranasal, oral, enteral, sublingual, rectal, intramuscular, intravenous, or subcutaneous means.
  • the compounds of this invention can be employed in combination with conventional excipients; i.e., pharmaceutically acceptable organic or inorganic carrier substances suitable for parenteral or enteral application which do not deleteriously react with the active compounds.
  • suitable pharmaceutically acceptable carriers include, but are not limited to, water, salt solutions, alcohol, vegetable oils, polyethylene glycols, gelatin, lactose, amylose, magnesium stearate, talc, silicic acid, viscous paraffin, perfume oil, fatty acid monoglyce ⁇ des and diglyce ⁇ des, pethroethral fatty acid esters, hydroxymethylcellulose, polyvinylpyrrolidone, etc.
  • the pharmaceutical preparations can be sterilized and if desired, mixed with auxiliary agents, e.g., lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts for influencing osmotic pressure, buffers. colorings, flavoring and/or aromatic substances and the like which do not deleteriously react with the active compounds.
  • auxiliary agents e.g., lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts for influencing osmotic pressure, buffers. colorings, flavoring and/or aromatic substances and the like which do not deleteriously react with the active compounds.
  • suitable vehicles consist of solutions, preferably oily or aqueous solutions, as well as suspensions, emulsions, or implants, including suppositories.
  • Ampules are convenient unit dosages.
  • tablets dragees or capsules having talc and/or a carbonhydrate carrier binder or the like, the carrier preferably being lactose and/or corn starch and/or potato starch.
  • a syrup, elixir or the like can be used wherein a sweetened vehicle is employed.
  • Sustained release composition can be formulated including those wherein the active component is protected with differentially degradable coatings, e.g., by microencapsulation, multiple coatings, etc.
  • a "therapeutically effective amount" of a pharmaceutical composition is an amount which is sufficient to achieve the desired pharmacological effect.
  • the dosage required to provide an effective amount of the composition will vary, depending upon the age, health, physical condition, sex, weight and extent of disease, of the recipient Additionally, the dosage may be determined by the frequency of treatment and the nature and scope of the desired effect.
  • SNOAC S-nitroso-N-acetylcysteine
  • a sample of S-nitroso-oxyhemoglobin was also treated with dithionitrite.
  • S-nitrosylation was verified, using standard methods for detection of S- nitrosothiols (Saville, Analyst, 83:670-612 (1958)). The Saville method, which assays free NO.
  • Figure 2 shows spectra of hemoglobin which has been incubated with S-nitroso- N-acetyl cysteine (SNOAC).
  • Tracing 5 reflects the product of treating the S-nitroso-oxyhemoglobin, for example, from tracing 1 , with dithionitrite.
  • the peaks at 350 nm are indicative of the presence of S-nitrosothiols.
  • the peaks at 400-450 nm are known as Soret peaks and they are indicative of the oxidative state of hemoglobin.
  • the small doublets at about 550 nm show that O 2 is binding at the hemoglobin metal center and that, therefore, its oxygen binding capacity is unchanged.
  • Tracing 5 shows a plateau, rather than a doublet, at this wavelength. This is because dithionitrite deoxygenates oxyhemoglobin and breaks down S- nitrosothiols to NO. NO disrupts O 2 binding, whereas S-nitrosothiols do not. This plateau indicates the shift from oxyhemoglobin to nitrosylhemoglobin.
  • S-nitrosylation of hemoglobin does not result in the formation of methemoglobin and consequent impairment in hemoglobin-oxygen binding. Furthermore, an additional experiment demonstrated that S-nitrosylation of hemoglobin causes a leftward shift in the hemoglobin-oxygen association curve, indicating an increase in oxygen binding. Thus, the reaction between S-nitrosothiols and hemoglobin not only eliminates the inhibition of oxygen binding which occurs from the reaction with NO», but actually increases binding and oxygenation of the blood.
  • S-nitrosothiols are important intermediates in the metabolism of organic nitrates and endogenously-derived NO. Furthermore, these compounds provide greater stability, a longer half life than NO, and retain its cyclic GMP-dependent bioactivity in blood vessels.
  • S-NO-hemoglobin may also be used to modulate the delivery of carbon monoxide or nitric oxide (bound to hemoglobin) to bodily tissues.
  • Hemoglobin is S-nitrosated by incubation with an alkylnitrite (e.g. , amyl nitrite or tert-butyl nitrite) or a thionitrite (e.g., S-nitroso-glutathione, S-nitroso- penicillamine, S-nitroso-cysteine and S-nitroso-N-acetylcysteine) for periods of one minute to one hour.
  • the pH is optimized within the range of 6.5 to 7.5 to achieve stoichiometric S-nitrosylation.
  • the reactions are preferably performed anaerobically. It may also be desired to perform the above procedure in saturating carbon monoxide solutions and under increased atmospheric pressures (i.e., 5 atmospheres) to prevent interaction of NO with the heme site.
  • Figure 4 shows the UV spectrum of S-NO-hemoglobin (Hb).
  • the spectra show that oxygen binding to the heme site is largely unaffected at S-NO/Hb stoichiometries (0.05 and 0.37) that are lacking in smooth muscle contractile activity. Higher ratios of S-NO/Hb (1.59) are associated with methemoglobin formation. Curves for oxy-Fe(II)-hemoglobin (dithionite-treated) and methemoglobin (K 3 Fe(CN)-treated) are shown for comparison.
  • Synthesis of S-NO-hemoglobin is monitored by three complementary methods, two of which were developed in this laboratory: photolysis- chemiluminescence and capillary electrophoresis methodologies.
  • the ligand binding to heme is determined spectrophoto metrically, using published extinction coefficients.
  • the structure of S-nitrosylated hemoglobin is then further characterized using SDS- polyacrylamide gel electrophoresis and isoelectric focusing immunoelectrophoresis.
  • Thephotolysis-chemiluminescence technique can be used to measure both protein and low molecular weight RS-NO.
  • the sample protein is introduced directly or as a chromatographic effluent from an attached FPLC/HPLC (for separation and identification of protein and amino acid RS-NO, respectively) into a photolysis cell where it is irradiated with a 200 watt mercury vapor lamp, designed to result in complete photolysis of the S-N bond. NO is then carried in a stream of helium towards the reaction chamber which yields the chemiiuminescence signal.
  • HgCl 2 which selectively displaces NO from thiol groups. We have confirmed that metal-nitrosyl complexes are not affected by this treatment.
  • the capillary electrophoresis method offers the added advantage of being able to separate hemoglobin variants as well as various RS-NO from their respective thiols and disulfides.
  • thoracic aorta, trachea, or intestinal tissues are isolated from anesthetized New Zealand white female rabbits. The tissues are cleaned, cut into small rings and mounted on stirrups connected to force transducers by which changes in isometric tension are recorded. Sustained contractions are elicited with agonists (i.e. , histamine or acetylcholine for airway smooth muscle or phenylephrine in the case of blood vessels), or by electrical field stimulation.
  • agonists i.e. , histamine or acetylcholine for airway smooth muscle or phenylephrine in the case of blood vessels
  • FIG. 5 shows reversal of hemoglobin (Hb)-induced contraction of aortic rings by S-nitrosylation.
  • Hemoglobin is shown to constrict vessels in a dose-dependent manner (•).
  • S-NO-Hb with a stoichiometry of 0.1 S-NO ⁇ Hb, entirely prevents the constrictor response ( o ).
  • Increasing the S-NO-Hb ⁇ Hb stoichiometry to 1 converts hemoglobin into a potent vasodilator ( ⁇ ).
  • S-NO-Hb ⁇ Hb stoichiometry to 1 converts hemoglobin into a potent vasodilator ( ⁇ ).
  • S- nitrosylation of hemoglobin abrogates the contraction induced by the native protein.
  • Measurement of highly precise oxygen binding curves is performed using standard tonometry, an Imai cell and/or a thin-layer Gill cell modified with an oxygen electrode.
  • a thin layer dual-beam method is used for studies of oxygen binding by intact red blood cells, with or without added cell-free hemoglobins.
  • a modified Hemoscan American Instruments Co. operated in a discontinuous mode to avoid dynamic error.
  • Rabbits are routinely used in the laboratory for assessment of hemodynamic responses to pharmacological agents.
  • Our objectives were, therefore, to demonstrate vasorelaxant and bronchodilator properties of S-NO-hemoglobin, determine the biological half life of the molecule and obtain insight into the degree of S-nitrosation required to overcome its contractile effects.
  • the data obtained in ring studies in vitro indicates that very small degrees of S-nitrosation are required to ameliorate vasoconstriction. Specifically, one NO per 40 heme groups is sufficient to prevent aortic smooth muscle contraction.
  • New Zealand white rabbits were anesthetized with ketamine (50 mg/kg i.m.) and sodium barbital (5-10 mg/kg i.v.) after which the femoral artery was cannulated to allow continuous measurement of blood pressure. Mean arterial pressure was then measured in response to intravenous bolus injections of S-NO hemoglobin (1 nmol/kg/min to 1 ⁇ m/kg/min) as well as continuous infusions. Blood pressure did not rise whereas native hemoglobin caused a hypertensive effect.

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Abstract

Cette invention concerne une hémoglobine d'origine humaine dans laquelle au moins une fraction thiol d'acide aminé est S-nitrosylée, et notamment une hémoglobine obtenue par nitrosylation d'une fraction thiol sur l'unique cystéine incluse dans la séquence de l'une des (ou des deux) sous-unité(s) β de l'hémoglobine. Cette invention se rapporte également à des compositions pharmaceutiquement acceptables qui contiennent de l'hémoglobine S-nitrosylée ainsi qu'à des utilisations de cette hémoglobine.
PCT/US1996/017508 1995-11-13 1996-10-31 S-nitroso-hemoglobine et ses utilisations therapeutiques WO1997018000A1 (fr)

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Cited By (8)

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EP0979073A1 (fr) * 1997-03-31 2000-02-16 The Children's Medical Center Corporation Nitrosylation effectuee pour inactiver des enzymes apoptotiques
EP1029551A2 (fr) * 1999-02-08 2000-08-23 Hokkaido University Complexe d'un metabolite de monoxyde d'azote - polyoxyalkylène-hemoglobine
US6627738B2 (en) 1995-09-15 2003-09-30 Duke University No-modified hemoglobins and uses therefor
US6855691B1 (en) 1995-09-15 2005-02-15 Duke University Methods for producing and using S-nitrosohemoglobins
US6884773B1 (en) 1995-09-15 2005-04-26 Duke University Modified hemoglobins, including nitrosylhemoglobins, and uses thereof
US6911427B1 (en) 1995-09-15 2005-06-28 Duke University No-modified hemoglobins and uses therefore
EP2094292A2 (fr) * 2006-10-23 2009-09-02 Ikor, Inc. Hémoglobine tétramère réticulée bloquant l'oxyde nitrique
WO2021198336A1 (fr) * 2020-04-02 2021-10-07 Hemarina Utilisation d'hémoglobine d'annélides pour traiter le syndrome de détresse respiratoire aiguë

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Cited By (16)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6911427B1 (en) 1995-09-15 2005-06-28 Duke University No-modified hemoglobins and uses therefore
US7538193B2 (en) 1995-09-15 2009-05-26 Duke University NO-modified hemoglobins and uses therefor
US7202340B2 (en) 1995-09-15 2007-04-10 Duke University No-modified hemoglobins and uses therefor
US6627738B2 (en) 1995-09-15 2003-09-30 Duke University No-modified hemoglobins and uses therefor
US6855691B1 (en) 1995-09-15 2005-02-15 Duke University Methods for producing and using S-nitrosohemoglobins
US6884773B1 (en) 1995-09-15 2005-04-26 Duke University Modified hemoglobins, including nitrosylhemoglobins, and uses thereof
EP0979073A1 (fr) * 1997-03-31 2000-02-16 The Children's Medical Center Corporation Nitrosylation effectuee pour inactiver des enzymes apoptotiques
EP0979073A4 (fr) * 1997-03-31 2004-04-07 Childrens Medical Center Nitrosylation effectuee pour inactiver des enzymes apoptotiques
US6667292B1 (en) 1999-02-08 2003-12-23 Hokkaido University Nitric monoxide metabolite-polyoxyalkylene-hemoglobin complex
EP1029551A3 (fr) * 1999-02-08 2001-03-07 Hokkaido University Complexe d'un metabolite de monoxyde d'azote - polyoxyalkylène-hemoglobine
EP1029551A2 (fr) * 1999-02-08 2000-08-23 Hokkaido University Complexe d'un metabolite de monoxyde d'azote - polyoxyalkylène-hemoglobine
EP2094292A2 (fr) * 2006-10-23 2009-09-02 Ikor, Inc. Hémoglobine tétramère réticulée bloquant l'oxyde nitrique
EP2094292A4 (fr) * 2006-10-23 2011-11-30 Ikor Inc Hémoglobine tétramère réticulée bloquant l'oxyde nitrique
US8129338B2 (en) 2006-10-23 2012-03-06 Ikor, Inc. Nitric oxide-blocked cross-linked tetrameric hemoglobin
WO2021198336A1 (fr) * 2020-04-02 2021-10-07 Hemarina Utilisation d'hémoglobine d'annélides pour traiter le syndrome de détresse respiratoire aiguë
FR3108842A1 (fr) * 2020-04-02 2021-10-08 Hemarina Utilisation d'hémoglobine d'Annélides pour traiter le syndrome de détresse respiratoire aiguë

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