WO2001068099A1 - Modulation of cardiovascular injury - Google Patents
Modulation of cardiovascular injury Download PDFInfo
- Publication number
- WO2001068099A1 WO2001068099A1 PCT/US2001/007806 US0107806W WO0168099A1 WO 2001068099 A1 WO2001068099 A1 WO 2001068099A1 US 0107806 W US0107806 W US 0107806W WO 0168099 A1 WO0168099 A1 WO 0168099A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- snmp
- administered
- heart
- tissue
- reperfusion
- Prior art date
Links
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/555—Heterocyclic compounds containing heavy metals, e.g. hemin, hematin, melarsoprol
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P7/00—Drugs for disorders of the blood or the extracellular fluid
- A61P7/04—Antihaemorrhagics; Procoagulants; Haemostatic agents; Antifibrinolytic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P9/00—Drugs for disorders of the cardiovascular system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P9/00—Drugs for disorders of the cardiovascular system
- A61P9/04—Inotropic agents, i.e. stimulants of cardiac contraction; Drugs for heart failure
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P9/00—Drugs for disorders of the cardiovascular system
- A61P9/06—Antiarrhythmics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P9/00—Drugs for disorders of the cardiovascular system
- A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
Definitions
- This invention relates to the heme oxygenase (HO) inhibitor tin- mesoporphyrin (SnMP) and its beneficial effects on the cardiovascular system.
- SnMP prevents the detrimental effects of free iron accumulation in tissue or organs in a mammal during ischemia and reperfusion or accompanying hemorrhage by inhibiting the release of free iron from heme in ischemic or hemorrhagic cardiac tissue.
- SnMP also acts as an antiarrhythmic agent by prolonging cardiac action potentials.
- the presence of metal ions, most importantly iron (Fe) is required in the generation of the oxygen-derived free radicals.
- Free hydroxyl radicals (*OH) are produced from O 2 and H 2 O 2 through the Haber- Weiss reaction catalyzed by iron.
- Ischemic myocardium is tissue particularly enriched with iron as a result of the leakage of red cells from damaged small blood vessels, their subsequent hemolysis and the degradation of their constituent hemoglobin.
- the degradation of heme is controlled by the enzyme heme oxygenase (HO), which is present and also induced in ischemic as well as hemorrhagic tissue.
- HO heme oxygenase
- Myoglobin and other heme proteins which are released from damaged muscle cells may also contribute heme as a substrate for HO and further add to the iron pool available for free radical production.
- HO has been found to have protective properties such as inhibiting inflammation, protecting endothelium and smooth muscle from oxidant stress, protecting tissues from heme induced injury (Platt & Nath, Nature Medicine, 1998, 4: 1364-1365) and protecting grafted tissue from chronic rejection (Hancock et al., Nature Medicine, 1998, 4: 1392-1396).
- HO activity also has toxic effects.
- a powerful oxidant heme degradation also produces bilirubin, implicated in tissue injury in jaundice and renal failure in hepatic disease.
- Another product of heme degradation is carbon monoxide (CO) which causes vasodilation.
- Iron chelating agents facilitate the binding and excretion of excess iron from tissue and thereby decrease free radical accumulation in tissue.
- Desferrioxamine has been used to limit infarct size and post-ischemic reperfusion injury (Bolli et al., Am J Physiol, 1987, 253:H1372-80).
- a major drawback of agents such as desferrioxamine as potential clinical pharmacologic agents in reducing reperfusion injury is that chelating agents are effective only after the iron has been released from heme and is already available for free radical production.
- chelating agents like desferrioxamine must be present in very high concentrations before they can satisfactorily lower iron levels.
- desferrioxamine typically must be administered continuously, i.e., via an infusion pump, to achieve results.
- Desferrioxamone is also known to produce allergic reactions in patients.
- the use of chelating agents for reducing reperfusion injury may be suitable in limited clinical environments, such as after surgery, however these agents are not practicable in situations of acute infarction or trauma.
- Antiarrhythmic drugs have been grouped together according to the pattern of electrophysiological effects that they produce and/or their presumed mechanisms of action.
- Class III antiarrhythmic drugs such as dofetilide, azimilide and amiodarone exert their effect by prolongation of the cardiac action potential (AP) (Groh WJ, et al. J. Cardiovasc. Electrophysiol. 1997, 8:529-536).
- AP cardiac action potential
- Reentrant rhythms in the heart are thought to be due to a spatial dispersion of repolarization and refractoriness (Restivo M, et al., Circ. Res. 1990, 66:1310-1327; Moe GK, Rev. Physiol. Biochem. Pharmacol. 1975, 72:55-81).
- the working hypothesis by which class III antiarrhythmic drugs operate is that prolongation of refractoriness increases the cycle length (CL) and thus the wavelength of potential reentrant wavelets.
- class III compounds including the two clinically available class III drugs sotalol and dofetilide, are methanesulfonimide compounds. It is believed that these act by blockade of the rapidly activating outward potassium current, IKr. The kinetics of blockade of this current is believed to be the cause of the reverse use- dependence. Regional differences in AP shape and duration are due to differences in ion channel densities and the ratio of ion channel subtypes.
- the APD gradient is uniform and oriented from apex to base and the total dispersion is less than 10 ms and not influenced by pacing site (Rosenbaum DS et al., Circulation, 1991, 84:1333-1345). Because of the minimal dispersion and the lack of nonuniform gradients, functional conduction block cannot be induced in the normal heart. However, other drugs which prolong APD can produce APD gradients due to regional differences in drug action. The neurotoxin anthopleurin-A can produce large APD gradients resulting in conduction block and reentry simulating characteristics of the long QT syndrome.
- Metalloporphyrins such as the tin (Sn) porphyrins, Sn-protoporphyrin (SnMP) and Sn-mesoporphyrin (SnMP), are known potent inhibitors of HO. They block binding of heme to HO, thus preventing the release of iron via heme catabolism. In contrast to iron chelating agents such as desferrioxamine, SnMP directly inhibits HO activity and prevents the degradation of heme and thus the release of its iron atom. SnMP has been used to inhibit HO activity in the intestine to reduce the absorption of iron from foodstuffs (U.S. Pat. No.
- HO inhibitors such as SnMP would be contra-indicated for treating the types of injuries resulting from post-ischemic reperfusion and hemorrhage, because, as discussed above, HO levels increase after ischemia, and the enzyme has been thought to play a protective role in reducing damage to surrounding tissue.
- present knowledge may be misleading since it does not take into account the toxic impact of free iron released at the site of injury.
- a method to prevent or inhibit the release and accumulation of free iron in reperfused tissue, rather than removing already formed excess free iron, during or soon after the onset of ischemia, to prevent or inhibit deleterious tissue injury resulting from peroxidation in reperfused and hemorrhagic tissue would be highly desirable.
- the identification of a class of cardio-active compounds that do not produce the deleterious side effects of known anti-arrhythmic agents, such as the class III antiarrhythmic drugs would also be desirable.
- the present invention addresses these needs in a wholly unexpected fashion.
- the present invention provides a method for preventing or reducing post- ischemic reperfusion and hemorrhagic injury associated with free iron formation in tissue.
- the method comprises administering a heme oxygenase (HO) inhibitor, preferably tin mesoporphyrin (SnMP), in an amount effective to inhibit HO activity.
- HO heme oxygenase
- SnMP tin mesoporphyrin
- the SnMP may be administered before, during or after an ischemic or hemorrhagic episode.
- the SnMP is administered as soon as ischemia or hemorrhaging is detected such that HO activity may be reduced or inhibited and free iron release is minimized or prevented.
- the SnMP is administered parenterally, intravenously, intramuscularly or directly into the affected tissue.
- the SnMP is administered in an amount effective to inhibit HO activity to reduce or inhibit free iron release and accumulation associated with reperfusion or hemorrhagic injury.
- the SnMP can be administered at a dosage of from 1 to 50 mg/kg body weight, preferably from 2 to 25 mg/kg body weight.
- the present invention further provides a method of inhibiting or reducing ventricular arrhythmia.
- the method comprises administering a heme oxygenase (HO) inhibitor, preferably tin mesopo ⁇ hyrin (SnMP), in an amount effective to prolong cardiac action potential duration (APD) and thereby preventing or reducing the occurrence of cardiac arrhythmia.
- HO heme oxygenase
- SnMP tin mesopo ⁇ hyrin
- the SnMP is administered for effective regulation of cardiac rhythm.
- SnMP may be administered as soon as arrythmia is detected, or for example, in an acute coronary event such as heart failure or congestive heart failure, and/or may be administrated prophylactically over a course to time to regulate cardiac rhythm.
- SnMP can also be used for treatment of acute ischemia and may be administered during or after an ischemic episode.
- the SnMP is administered parenterally, intravenously, intramuscularly or directly into the affected tissue.
- the SnMP is administered in an amount effective to prolong APD.
- the SnMP can be administered at a dosage of from 1 to 50 mg/kg body weight, preferably from 2 to 25 mg/kg body weight.
- Figure 1 shows the ECG tracing of a rabbit heart.
- the heart was perfused with oxygenated Tyrode 's solution; the left coronary artery was clamped for 3 minutes; the ligature was loosened and the heart was reperfused.
- the ECG recording was made during and for 10 minutes after re-perfusion.
- Figure 2 shows the ECG tracing of the rabbit heart treated as described in Figure 1, after which a 0.5g % hemoglobin in Tyrode 's solution was perfused into the heart.
- Figure 3 shows the ECG tracing of the rabbit heart treated as described in
- Figure 4 shows the ECG tracing of the rabbit heart treated as described in Figure 3 after perfusion for 15-20 minutes with hemoglobin/Tyrode's solution.
- Figure 5 shows a schematic diagram of the perfusion chamber and optics system utilized for the experiments described in the Examples.
- a Langendorff perfused heart is stained with the voltage sensitive dye di-4-ANEPPS.
- the epicardium is illuminated with light from 2 halogen light sources (LS) which passes through 520+20 nm interference filters. Fluoresced light from the epicardial surface is gathered through a lens, filtered at 645 nm, and focused onto a 124-element photodiode array (lxl mm/photodiode).
- LS halogen light sources
- Figure 6 shows the AP prolonging effect of SnMP in the normal guinea pig heart.
- the left panel (A) shows optical APs before and after administration of 50 ⁇ l SnMP at paced cycle lengths (CL) ranging from 250-450 ms.
- the right panel (B) shows a plot of APD as a function of CL before and after administration of SnMP.
- Optical APs are shown in control test (solid line) and in the presence of 50 ⁇ l SnMP (dotted line).
- Figure 7 shows the AP prolonging effect of SnMP in the normal rabbit heart.
- the left panel (A) shows optical APs before and after administration of lOO ⁇ l SnMP at paced cycle lengths (CL) ranging from 300-500 ms.
- the right panel (B) shows a plot of
- APD as a function of CL before and after administration of SnMP.
- Optical APs are shown in control test (solid line) and in the presence of lOO ⁇ l SnMP (dotted line).
- Figure 8 shows the long lasting effect of a single bolus of 50 ⁇ l SnMP administered in a perfused rabbit heart.
- the upper trace is the control.
- the middle trace is the APD at 5 minutes after SnMP administration.
- the lower trace is the APD at 30 minutes after SnMP administration.
- FIG. 9 shows that SnMP does not affect the activation pattern of normal cardiac rhythm.
- Activation maps are shown from a rabbit heart during basic paced rhythm. Each shaded zone represents an isochronal region activated at successive 1 ms intervals. Sj activation maps are shown for control and after 200 ⁇ l SnMP was administered in a rabbit heart.
- FIG 10 shows isochronal maps of APD distribution before and after administration of SnMP.
- Each shaded zone represents an isochronal region activated at successive 10 ms isochronal intervals during left ventricular (LV) pacing (400ms CL) of the rabbit heart.
- Panel A shows the control map, the LN AP and the ECG; Panel B shows these after administration of 100 ⁇ l SnMP.
- Figure 11 shows that SnMP prevents ischemia-induced APD shortening in the ischemic zone.
- Base sites are proximal to ligation site; apex sites are distal to ligation site.
- the left panel shows the control, no ligation; the middle panel shows that coronary occlusion shortens APD in ischemic zone (apex) after 5 minutes of ischemia; the right panel shows that SnMP prolongs APD in normal zone (base) and 5 minutes of myocardial ischemia does not result in appreciable APD shortening.
- the present invention is based on the discovery that the administration to animals of SnMP before, during, or after an ischemic cardiac episode inhibited or prevented the detrimental effects of ischemia and reperfusion.
- Experiments in a rabbit heart model showed that administration of SnMP prior to or following an ischemic event reduced, prevented and/or improved recovery from impaired myocardial function resulting from post-ischemic reperfusion.
- Administration of SnMP also inhibited deterioration of myocardial function following administration of hemoglobin directly to the heart during reperfusion.
- the administration of SnMP to a heart which showed heart block with limited ventricular beats improved the heart function such that regular sinus rhythm (RSR) was achieved and full recovery from post ischemic reperfusion was obtained. Subsequent infusion of hemoglobin did not alter the RSR.
- SnMP ameliorated re-perfusion injury that occurred following ischemia.
- SnMP pre- treatment reduced or inhibited iron toxicity following direct infusion of hemoglobin.
- the present invention is based on the discovery that SnMP administered to isolated animal hearts during basic paced rhythm demonstrated an anti-arrhythmic effect similar to that of current class-Ill anti-arrhythmic agents, however without the deleterious side effects accompanying the use of these agents such as reverse- use dependence. Applicants found that the overall APD gradient was maintained throughout the epicardium with the administration of SnMP and the degree of APD increase was relatively uniform in the epicardial layer. When SnMP was administered to a perfused guinea pig heart, prolonged APD was observed within five minutes of administration and was maintained for the duration of the study.
- SnMP acts as an antiarrhythmic agent and provides a means for restoring rhythmic contraction, selectively prolongs the action potential duration and concomitantly increases the refractory period of heart cells without significant effect on cardiac conduction. SnMP is therefore suitable for the treatment of mammals suffering from arrhythmic disorders or disease.
- Action potential is a response, elicited by stimulus in excitable cells, which is measured from the beginning of membrane depolarization (upstroke) to return to baseline potential (repolarization).
- Action potential duration is the time interval between the upstroke of the action potential and its point of repolarization.
- Arrhythmia is any variation from the normal rhythm of the heartbeat, including, without limitation, sinus arrhythmia, premature heartbeat, heartblock, fibrillation, flutter, tachycardia, and premature ventricular contractions.
- Arrhythmia occurs, for example, by deprivation of oxygen and other blood constituents in the myocardium. Production of oxygen derived free radicals during reperfusion also causes arrhythmia. Particularly dangerous is ventricular fibrillation whereby the heart loses its ability to contract in a coordinated fashion needed to pump blood properly.
- Reperfusion is the return of the flow of blood through vessels, e.g., from an artery, through the vasculature to and over the bodily tissues after an episode of reduced blood flow, for example, ischemia.
- Reperfusion also occurs in transplanted organs; acute transplantation rejection, e.g., involving a complement mediated cascade, occurs from an immediate reaction after reperfusion.
- Reperfusion also occurs in cases of massive trauma or "crush injury” where limbs or other body areas may be crushed or mangled such as occurs in moving vehicle accidents and construction site accidents.
- Reperfusion causes tissue damage due in part to the generation of free hydroxyl radicals. Free hydroxyl radicals damage cell membranes via lipid peroxidation, and the degradation of DNA and proteins.
- Ischemia is a localized decrease in blood supply to an organ due to obstruction or constriction of a blood vessel due, for example, to a blood clot or other obstruction.
- the decrease in blood supply may result in stroke (if the organ is the brain), heart attack (if the organ is the heart) or mechanical organ failure.
- Ischemia can occur in an organ, tissue or parts thereof.
- necrosis death of cells or tissues can occur.
- necrosis can also occur as a result of reperfusion by the action of, inter alia, free hydroxyl radicals.
- Infarct is an area of tissue that undergoes necrosis as a result of an obstruction of blood supply to the tissue due, for example, to a blood clot or other obstruction in the heart or blood vessel.
- a "myocardial infarction” is necrotic heart tissue.
- Hemorrhage is the release or escape of blood from the blood vessels into tissue and is typically manifested by an excessive loss of blood. Hemorrhages can occur as a result of traumatic injury, e.g., an automobile accident, or as the result of an infectious disease, e.g., as in hemorrhagic fever. Bruising, whether from trauma or surgery, is a localized hemorrhage (hematoma).
- Certain synthetic metalloporphyrins are HO inhibitors.
- Such "compounds include metalloprotoporphyrins and metallomesoporphyrins wherein the metal group may be selected from tin, chromium, platinum, zinc, cobalt, nickel, copper, silver and manganese or other elements.
- the ring tetrapyrrole may be altered as well - viz proto- versus meso- porphyrins.
- Metallopo ⁇ hyrins may be obtained commercially or may be synthesized by described methods, see, e.g. U.S. Pat. No. 4,692,440 for synthesis of
- SnMP which has received an IND from the FDA for treating hyperbilirubinemia in neonates, is preferred.
- the present invention further provides other HO inhibitors useful for treatment of conditions arising from heme degradation by HO.
- Inhibitors that are contemplated include fragments, peptides, nucleic acids and oligonucleotides, carbohydrates, phospholipids and other lipid derivatives, steroids and steroid derivatives, antisense nucleic acids (including ribozymes) which may be used to inhibit expression of HO, anti-HO antibodies, small molecule inhibitors and vitamin B 12 and its derivatives.
- Conditions which may be treated with an HO inhibitor according to the present invention include hemorrhagic injury, which may result from surgery or other trauma to body tissue, reperfusion injury which may occur following resumption of blood flow in blood vessels following loosening or removal of blood clots either chemically (e.g., using tissue plasminogen activator) or mechanically (angioplasty), or resumption of blood flow after bypass surgery, or stroke, or other acute cardiac episodes; treatment during infarction, in order to minimize the damage to tissues surrounding the infarct area.
- the HO inhibitor is provided prior to an elective surgery, e.g., plastic surgery of the face, to prevent extravasation associated with bruising.
- the HO inhibitors can be used to treat or regulate arrhythmia in patients in need thereof.
- the present invention can be used as an effective first aid treatment for ischemic conditions or acute coronary events such as myocardial infarction or stroke, heart failure, particularly congestive heart failure, preferably when the HO inhibitor is a safety-tested compound like SnMP.
- emergency personnel emergency medical technicians, firefighters, police
- other responsible persons flight attendants, conductors, event ushers, etc.
- suspect an infarction or observe symptoms such as chest pain, shortness of breath, fatigue, and anxiety
- a bolus of HO inhibitor can be administered.
- the HO inhibitor chosen for such emergency use does not have any adverse side-effects, it will not be necessary for the emergency personnel to make or confirm a diagnosis.
- This invention provides a valuable adjunct to infarct therapies, including but not limited to, administration of aspirin (acetylsalicylic acid) to prevent or inhibit clot formation, and administration of a "clot-busting" agent such as tissue plasminogen activator or streptokinase. It is also ideally used in combination with balloon angioplasty and other techniques that temporarily block blood flow.
- aspirin acetylsalicylic acid
- a "clot-busting" agent such as tissue plasminogen activator or streptokinase. It is also ideally used in combination with balloon angioplasty and other techniques that temporarily block blood flow.
- HO inhibitors like SnMP, are highly useful as components of perfusion solutions, e.g., for heart-lung machines during bypass surgery, or for infusing organs prior to transplantation.
- HO inhibitors can be administered to a subject prior to and during perfusion of the heart and lungs after completion of the surgery requiring use of a heart-lung machine, and prior to perfusion of a transplanted organ after grafting it into the host.
- the HO inhibitors described herein can be administered to an individual in need of treatment as a therapeutically effective dose in a pharmaceutically acceptable carrier.
- pharmaceutically acceptable refers to molecular entities and compositions that are physiologically tolerable and do not typically produce an allergic or similar untoward reaction, such as gastric upset, dizziness and the like, when administered to a human.
- pharmaceutically acceptable means approved by a regulatory agency of the Federal or a state government or listed in the U.S. Pharmacopeia or other generally recognized pharmacopeia for use in animals, and more particularly in humans.
- carrier refers to a diluent, adjuvant, excipient, or vehicle with which the compound is administered.
- Such pharmaceutical carriers can be sterile liquids, such as water and oils, including those of petroleum, animal, vegetable or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil and the like.
- Water or aqueous solution saline solutions and aqueous dextrose and glycerol solutions are preferably employed as carriers, particularly for injectable solutions. Suitable pharmaceutical carriers are described in "Remington's Pharmaceutical Sciences” by E.W. Martin (18th Edition).
- a therapeutically effective amount is used herein to mean an amount sufficient to reduce by at least about 15 percent, preferably by at least 50 percent, more preferably by at least 90 percent, and most preferably prevent, a clinically significant deficit in the activity, function and response of the host.
- a therapeutically effective amount is sufficient to cause an improvement in a clinically significant condition in the host.
- a therapeutically effective amount of an HO inhibitor is an amount effective to inhibit HO activity in reperfused or hemorrhagic tissue.
- Another example of a therapeutically effective amount of an HO inhibitor such as SnMP is an amount effective to prolong cardiac action potential duration (APD) in a patient suffering from arrhythmia.
- the actual dosage regimen (amount and frequency) of an HO inhibitor will be determined by the skilled physician, based on the condition, the age, sex, weight, and health of the patient, and other factors well within the ordinary level of skill in the art.
- the dosage range for SnMP employed for decreasing the rate of formation of free iron in reperfused blood or tissue is approximately from 1 to 50 mg/kg body weight, preferably 2 to 25 mg/kg body weight.
- the concentration of SnMP can be 2 to 25 g/liter to provide a dosage of from 2 to 25 mg/kg body weight.
- the single dosage units will typically contain from 2 mg/ml to 25 mg/ml of SnMP solution.
- compositions according to the present invention comprising HO inhibitors may be administered through various modes, for example, as described for metallopo ⁇ hyrins. See U.S. Pat. No. 4,692,440 and U.S. Pat. No. 4,657,902 (parenteral administration to increase the rate at which heme is excreted), U.S. Pat. No. 4,619,923 (parenteral administration to increase the rate of tryptophan metabolism in liver), U.S. Pat.
- compositions according to the present invention may be included as part of a kit, such as a medical emergency kit.
- a kit may itself be a component of an emergency first aid kit accompanying a portable defribillator, such as are presently used to equip police cars, ai ⁇ lanes, etc.
- a therapeutically effective amount of a HO inhibitor e.g., SnMP
- SnMP a HO inhibitor
- the composition may be packaged in a container, preferably a syringe or other injection device, preferably in a pre-measured unit dosage form.
- the compound SnMP is known to be stable in solution at room temperature for several years.
- the model system used for the experiments in Examples 1-3 was the Langendorff-perfused rabbit heart preparation. Surgical procedures were performed as previously described (Salama G et al., Am. J. Physiol 1987, 252:H384-H394).
- rabbits (approximate weight 2-4 kg, of either sex) were anesthetized by intraperitoneal injection of sodium phenobarbital and heparinized.
- a mid-thoracotomy was performed and the hearts were rapidly excised and placed in cold oxygenated Tyrode 's containing 100 U/ml heparin.
- the excised hearts were rapidly annulated at the aorta and retrogradely perfused in a modified Langendorff apparatus.
- the Tyrode 's solution comprised (in mM): NaCl 130, KC1 4.75, CaCl 2 1.0, MgSO 4 1.2, NaHCO 3 12.5, and glucose 15.0.
- the solution was continuously bubbled with 95%-O 2 / 5%CO 2 through a fitted glass tube. Temperature was maintained at 36 +0.3C by monitoring the temperature of the efusate within the closed chamber. A small incision was made in the pulmonary artery for drainage. A variable speed roller. pump was used to maintain reperfusion pressure of 80 to 90 cm H 2 O using a perfusion flow rate of 2.0ml/min/g heart weight.
- a custom designed perfusion chamber was used to study the isolated rabbit hearts (Salama G et al., 1987).
- the heart in the chamber was immersed in perfusate.
- Bipolar surface electrocardiograms were recorded using Teflon-coated silver wires. Recording and stimulating electrodes were positioned on the epicardial surfaces or perfusion chamber side pads.
- Hearts were paced at a fixed cycle length SI ranging from 200 to 1200 ms with bipolar Ag-AgCl electrodes placed at the left ventricular free wall and the right ventricle near the anterior septum.
- Ventricular stimulation was applied using constant current pulses of 2.5ms duration at 1.5 times diastolic threshold.
- Steady state APD measurements were determined after a minimum of 30 beats.
- the preparation was perfused with a 0.5 gm% hemoglobin solution in Tyrode's.
- the ECG showed only small amplitude QRS cycles.
- the heart was non-functional within 5 minutes after hemoglobin perfusion.
- the ECG tracing is shown in Figure 2. Subsequently the heart was reperfused with Tyrode's solution alone. After 2- 5 minutes, ventricular rhythm returned to normal sinus rhythm. The heart was then perfused with Tyrode's solution containing 20 ⁇ mol/L SnMP for 5-10 minutes to allow full exposure of the cardiac tissue to the SnMP. The coronary artery was then ligated for three minutes after which the ligature was loosened (reperfusion). The ECG was recorded during and for ten minutes after reperfusion. No rhythm changes were observed. The ECG tracing is shown in Figure 3. After another 15-20 minutes the heart was again perfused with 0.5% hemoglobin Tyrode's solution. No deleterious effect on the ECG was recorded; the heart demonstrated normal sinus rhythm (NSR). The ECG tracing is shown in Figure 4.
- the epicardial surface of the heart was illuminated with light from two 45 W tungsten halogen lamps (LS). The light was collimated and passed through 520+20 nm interference filters. A 45° mirror in the optical apparatus was used to focus the grid pattern on the region of interest using a 35 mm camera lens (50 mm, Fl : 1.4, Nikon). Epi-fluorescent light from the stained heart was gathered through a lens (L), projected through a 645 nm cutoff filter, and focused to form an image of the heart on a 12x12 element photodiode array. The photodiode array consisted of 144 square diode elements, with each diode having dimensions of 1.0 x 1.0 mm separated by 0.1 mm. 124 diodes were current to voltage converted and sampled. The depth of field of the optics was approximately 150 ⁇ m.
- Hearts were stained by gradual injection of 40 to 60 ⁇ l from a 2.5 mM stock solution of dye into a 5ml bubble trap situated directly above the aortic cannula. The final dye concentration was approximately 1.8 ⁇ M; 10 to 15 minutes was allowed for the staining to be completed. The procedure resulted in homogeneous staining throughout the heart because the dye was efficiently delivered via coronary vessels. For longer protocols for which photobleaching (prolonged exposure of the dye to light degrades its usefulness) and/or dye washout may reduce the optical signal amplitudes, hearts were restained with smaller amounts of dye (5-10 ml) to restore the original signal-to-noise ratio.
- Each data acquisition epoch comprised a scan of 128 simultaneously recorded traces (124 photodiodes plus 4 instrumentation channels).
- the multiplexed instrumentation channels monitored the stimulus pulses and surface electrogram signals from the ventricle.
- the duration of each stored acquisition epoch ranged from 1-15 seconds.
- the photodiode currents from each of the 124 sites were fed to a current-to- voltage converter, amplified, and high pass filtered to remove background fluorescence.
- the filtered signals were further amplified using a selectable gain amplifier stage then digitized with 12-bit resolution with a sampling rate of 0.64 ms/channel (1.6 kHz) using a Microstar A/D converter board in an IBM compatible PC computer.
- a custom designed analysis system (IDL 4.0, Research Systems, Inc; Boulder, CO) was used for detection of activation and repolarization times at each site in the array.
- Activation time was defined as the peak temporal derivative of the AP upstroke and recovery was defined as the point of maximum second derivative during repolarization. A good concordance has been previously shown between the latter value and the refractory period of normal guinea pig myocardium (Efimov et al 1994). Animals The studies were performed in 9 guinea pig hearts and 13 rabbit hearts. For study of the electrophysiologic effect of SnMP in normal myocardium, each animal served as its own control. Steady-state rate analysis was performed in those experiments in which high fidelity signals were obtained in control and after SnMP and with a 1:1 activation pattern at all cycle lengths.
- Rabbit Model Surgical Preparation
- the rabbit model has been extensively described (Gillis AM, et al., Am.J.Physiol 1996, 271:H784-H789).
- the electrophysiologic properties of the ionic currents that constitute the rabbit AP have been well characterized and may better represent the human heart rather than smaller animals.
- the dimensions of the rabbit heart are well suited for the spatial resolution of the optical system and permit imaging of large planar surfaces of the heart and specific cardiac structures.
- the optical resolution of the system can be adjusted from 400-1800 mM/pixel, depending on the dimension of the region of interest.
- Electrophysiologic observations can be correlated at high resolution with the anatomic features of the heart.
- New Zealand young rabbits of either sex were anesthetized by intravenous injection of fentanyl citrate (100 mg/kg + 15 mg/kg/hr) and heparinized (1000 U/kg).
- a tracheotomy was performed on each animal and the animal was intubated with an endotracheal tube.
- the rabbit was ventilated with room air via a positive pressure ventilator (MD Industries, Mobile, AL).
- a midline thoracotomy was performed and the heart was exposed in a pericardial cradle.
- the heart was rapidly excised and placed in cold oxygenated Tyrode's solution, containing 1 ,000 U/l heparin.
- the heart was then cannulated and retrogradely perfused in a modified Langendorff setup and optical perfusion chamber.
- the hearts were perfused with oxygenated (95% O 2 , 5% CO 2 ) modified
- a ligature was placed around the left anterior descending artery using a 5-0 suture with a non-cutting needle.
- a snare comprising a small piece of polyethylene tubing and a small hemostat was used to reversibly ligate the coronary artery.
- SnMP had a similar effect in the rabbit heart.
- the shape of the rabbit AP is different from that of the guinea pig and may be related to differences in the density and ratios of the delayed rectifier repolarizing currents, IK,, and IK r (the slow and rapid activating components, respectively).
- IK, and IK r the delayed rectifier repolarizing currents
- a dose of 100 ⁇ l SnMP was administered.
- Figure 7 As in the guinea pig, SnMP prolonged APD at all CLs and the amount of prolongation was slightly greater than in the guinea pig. The mean increase was 41.4 + 1.5 ms. Within the CL range used in this protocol, there appeared to be no reverse use dependence associated with SnMP.
- Figure 8 shows that the effect of a single bolus of SnMP is long lasting. SnMP delivered as a single bolus dose of 50 ⁇ l prolonged APD in a guinea pig heart within 5 minutes ( Figure 8, middle trace). The effect was maintained for the duration of the experiments. The lower trace in Figure 8 shows that the APD at 30 minutes was similar to that observed 5 minutes after the administration of SnMP.
- Activation Mapping Isochronal maps of APD were constructed to assess the spatial organization of APD prolongation by SnMP in the heart (Salama G et al. , Am. J. Physiol 1987, 252.H384-H394). Activation maps during basic paced rhythm were obtained. In the maps shown in Figure 9, each shaded zone represents an isochronal region activated at successive 1 ms intervals. Activation maps for the basic paced rhythm S j are shown for control and after 200 ⁇ l SnMP was administered in a rabbit heart. Hearts were paced from the right ventricle. The shape of the isochrones and total activation time were similar before and after SnMP and indicated a lack of effect on conduction in the heart.
- the APD maps shown in Figure 10 were drawn at 10 ms isochronal intervals. Representative optical APs from the 124 recordings made are shown below each map along with an ECG.
- A normal heart
- APD the normal heart
- B overall APD in the imaged area
- B the dispersion of APD
- the drug increased APD but did not cause gradients of APD dispersion to occur, which are potentially arrhythmogenic.
- Examples 3-10 provide a number of clinical circumstances in which the immediate administration of a dose of HO inhibitor such as SnMP may prove useful in ameliorating the damaging sequelae of heme-iron tissue injury.
- a single dose, or multiple doses (at approximately 24 hours apart) of the inhibitor may be administered at the slightest indication of need since the inhibitor has been shown to be innocuous in human newborns subjects for other pu ⁇ oses.
- Organs to be used for transplantation undergo reperfusion injury when implanted into recipients.
- the solutions in which the organs are maintained should contain SnMP in a concentration range of 1-20 ⁇ mol/liter.
Abstract
Description
Claims
Priority Applications (4)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
EP01916568A EP1261346A4 (en) | 2000-03-10 | 2001-03-12 | Modulation of cardiovascular injury |
CA002402645A CA2402645A1 (en) | 2000-03-10 | 2001-03-12 | Modulation of cardiovascular injury |
AU2001243578A AU2001243578A1 (en) | 2000-03-10 | 2001-03-12 | Modulation of cardiovascular injury |
JP2001566663A JP2003526668A (en) | 2000-03-10 | 2001-03-12 | Modulation of cardiovascular injury |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US18843400P | 2000-03-10 | 2000-03-10 | |
US60/188,434 | 2000-03-10 |
Publications (1)
Publication Number | Publication Date |
---|---|
WO2001068099A1 true WO2001068099A1 (en) | 2001-09-20 |
Family
ID=22693126
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/US2001/007806 WO2001068099A1 (en) | 2000-03-10 | 2001-03-12 | Modulation of cardiovascular injury |
Country Status (5)
Country | Link |
---|---|
EP (1) | EP1261346A4 (en) |
JP (1) | JP2003526668A (en) |
AU (1) | AU2001243578A1 (en) |
CA (1) | CA2402645A1 (en) |
WO (1) | WO2001068099A1 (en) |
Cited By (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2006506440A (en) * | 2002-11-20 | 2006-02-23 | ウェルスプリング・ファーマシューティカル・コーポレイション | Water-soluble mesoporphyrin compound and production method |
US7960371B2 (en) | 2006-10-04 | 2011-06-14 | Infacare Pharmaceutical Corporation | High-purity large-scale preparation of stannsoporfin |
US8178664B2 (en) | 2002-06-04 | 2012-05-15 | Infacare Pharmaceutical Corporation | Preparation of metal mesoporphyrin compounds |
US8735574B2 (en) | 2011-03-30 | 2014-05-27 | Infacare Pharmaceutical Corporation | Methods for synthesizing metal mesoporphyrins |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1991016053A1 (en) * | 1990-04-24 | 1991-10-31 | The Rockefeller University | The use of synthetic metalloporphyrins in the treatment of hypertension and other vascular disorders |
US6147070A (en) * | 1998-06-05 | 2000-11-14 | Facchini; Francesco | Methods and compositions for controlling iron stores to treat and cure disease states |
Family Cites Families (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4783470A (en) * | 1986-01-14 | 1988-11-08 | The Rockefeller University | Hematin derived anticoagulant |
US5162313A (en) * | 1989-03-16 | 1992-11-10 | The Rockefeller University | Control of heme and iron concentrations in body tissues |
US5948771A (en) * | 1996-01-31 | 1999-09-07 | The Trustees Of Columbia University In The City Of New York | Method for treating heart failure using tetrapyrroles and metallotetrapyrroles |
-
2001
- 2001-03-12 CA CA002402645A patent/CA2402645A1/en not_active Abandoned
- 2001-03-12 WO PCT/US2001/007806 patent/WO2001068099A1/en not_active Application Discontinuation
- 2001-03-12 JP JP2001566663A patent/JP2003526668A/en not_active Withdrawn
- 2001-03-12 EP EP01916568A patent/EP1261346A4/en not_active Withdrawn
- 2001-03-12 AU AU2001243578A patent/AU2001243578A1/en not_active Abandoned
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1991016053A1 (en) * | 1990-04-24 | 1991-10-31 | The Rockefeller University | The use of synthetic metalloporphyrins in the treatment of hypertension and other vascular disorders |
US6147070A (en) * | 1998-06-05 | 2000-11-14 | Facchini; Francesco | Methods and compositions for controlling iron stores to treat and cure disease states |
Non-Patent Citations (1)
Title |
---|
See also references of EP1261346A4 * |
Cited By (15)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US8178664B2 (en) | 2002-06-04 | 2012-05-15 | Infacare Pharmaceutical Corporation | Preparation of metal mesoporphyrin compounds |
JP2006506440A (en) * | 2002-11-20 | 2006-02-23 | ウェルスプリング・ファーマシューティカル・コーポレイション | Water-soluble mesoporphyrin compound and production method |
US9107927B2 (en) | 2006-10-04 | 2015-08-18 | Infacare Pharmaceutical Corporation | High-purity large-scale preparation of stannsoporfin |
US8530458B2 (en) | 2006-10-04 | 2013-09-10 | Infacare Pharmaceutical Corporation | High-purity large-scale preparation of stannsoporfin |
US8835416B2 (en) | 2006-10-04 | 2014-09-16 | Infacare Pharmaceutical Corporation | High-purity large-scale preparation of stannsoporfin |
US7960371B2 (en) | 2006-10-04 | 2011-06-14 | Infacare Pharmaceutical Corporation | High-purity large-scale preparation of stannsoporfin |
US9517239B2 (en) | 2006-10-04 | 2016-12-13 | Infacare Pharmaceutical Corporation | High-purity large-scale preparation of stannsoporfin |
US9902745B2 (en) | 2006-10-04 | 2018-02-27 | Infacare Pharmaceutical Corporation | High-purity large-scale preparation of stannsoporfin |
US10273255B2 (en) | 2006-10-04 | 2019-04-30 | Infacare Pharmaceutical Corporation | High-purity large-scale preparation of stannsoporfin |
US10662209B2 (en) | 2006-10-04 | 2020-05-26 | Mallinckrodt Hospital Products IP Limited | High-purity large-scale preparation of stannsoporfin |
US11078220B2 (en) | 2006-10-04 | 2021-08-03 | Mallinckrodt Hospital Products IP Limited | High-purity large-scale preparation of stannsoporfin |
US8735574B2 (en) | 2011-03-30 | 2014-05-27 | Infacare Pharmaceutical Corporation | Methods for synthesizing metal mesoporphyrins |
US9181285B2 (en) | 2011-03-30 | 2015-11-10 | Infacare Pharmaceutical Corporation | Methods for synthesizing metal mesoporphyrins |
US9688705B2 (en) | 2011-03-30 | 2017-06-27 | Infacare Pharmaceutical Corporation | Methods for synthesizing metal mesoporphyrins |
US10533024B2 (en) | 2011-03-30 | 2020-01-14 | Mallinckrodt Hosptial Products Ip Limited | Methods for synthesizing metal mesoporphyrins |
Also Published As
Publication number | Publication date |
---|---|
EP1261346A1 (en) | 2002-12-04 |
CA2402645A1 (en) | 2001-09-20 |
EP1261346A4 (en) | 2004-10-13 |
JP2003526668A (en) | 2003-09-09 |
AU2001243578A1 (en) | 2001-09-24 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
US4673563A (en) | Adenosine in the treatment of supraventricular tachycardia | |
Bentley et al. | Vascular effects of propofol: smooth muscle relaxation in isolated veins and arteries | |
Jia et al. | Cold injury to nerves is not due to ischaemia alone. | |
Hong et al. | Magnolol reduces infarct size and suppresses ventricular arrhythmia in rats subjected to coronary ligation | |
US4364922A (en) | Adenosine antagonists in the treatment and diagnosis of A-V node conduction disturbances | |
CA2061270C (en) | Method and composition for treating reperfusion injury | |
Wilson et al. | Serious ventricular dysrhythmias after intracoronary papaverine | |
JP2002518983A (en) | How to adjust the vagus tone | |
DiPALMA et al. | Antifibrillatory drugs | |
JP3623501B2 (en) | Treatment of neurological conditions with interleukin-1 inhibitory compounds | |
JP3253302B2 (en) | Parenteral solution containing 3-dialkylaminoethoxybenzoyl-benzofuran | |
CH683966A5 (en) | Compounds of the ftalidrazidici class as active substances in anti-hypoxic agents and defense. | |
EP1261346A1 (en) | Modulation of cardiovascular injury | |
Pugsley et al. | Protamine is a low molecular weight polycationic amine that produces actions on cardiac muscle | |
Yilmaz et al. | CDP-choline prevents cardiac arrhythmias and lethality induced by short-term myocardial ischemia–reperfusion injury in the rat: involvement of central muscarinic cholinergic mechanisms | |
Klein et al. | Antiinflammatory agent BW 755 C in ischemic reperfused porcine hearts | |
Chan et al. | Trilinolein reduces infarct size and suppresses ventricular arrhythmias in rats subjected to coronary ligation | |
Reele et al. | The effects of continuous infusions of prostacyclin-Na (epoprostenol-sodium) on platelet counts, ADP-induced aggregation, and cyclic AMP levels in normal volunteers | |
Ducko et al. | Potassium channel openers: are they effective as pretreatment or additives to cardioplegia? | |
JP3193767B2 (en) | Drugs for regeneration of neuronal defects and restoration of energy metabolism of nerve cells after interruption of blood circulation | |
RU2168993C1 (en) | Agent for treatment of patients with nonstable stenocardia and acute myocardial infarction | |
Lazar et al. | Beneficial effects of angiotensin-converting enzyme inhibitors during acute revascularization | |
US4495201A (en) | Pharmaceutical composition and method for treating heart diseases therewith | |
RU2350326C2 (en) | Use of taliporfin or its derivatives in treatment of heart diseases, and also their derivation | |
RU2783444C1 (en) | Cardioprotective and antiarrhythmic medicine |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
AK | Designated states |
Kind code of ref document: A1 Designated state(s): AE AG AL AM AT AU AZ BA BB BG BR BY BZ CA CH CN CO CR CU CZ DE DK DM EE ES FI GB GD GE HR HU ID IL IN IS JP KE KG KP KR KZ LC LK LR LS LT LU LV MA MD MG MK MN MW MX MZ NO NZ PL PT RO RU SD SE SG SI SK SL TJ TM TR TT TZ UA UG US UZ VN YU ZA ZW |
|
AL | Designated countries for regional patents |
Kind code of ref document: A1 Designated state(s): GH GM KE LS MW MZ SD SL SZ TZ UG ZW AM AZ BY KG KZ MD RU TJ TM AT BE CH CY DE DK ES FI FR GB GR IE IT LU MC NL PT SE TR BF BJ CF CG CI CM GA GN GW ML MR NE SN TD TG |
|
121 | Ep: the epo has been informed by wipo that ep was designated in this application | ||
DFPE | Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101) | ||
WWE | Wipo information: entry into national phase |
Ref document number: 2001243578 Country of ref document: AU |
|
ENP | Entry into the national phase |
Ref document number: 2001 566663 Country of ref document: JP Kind code of ref document: A |
|
WWE | Wipo information: entry into national phase |
Ref document number: 2001916568 Country of ref document: EP |
|
WWE | Wipo information: entry into national phase |
Ref document number: 2402645 Country of ref document: CA Ref document number: PA/A/2002/008843 Country of ref document: MX |
|
WWP | Wipo information: published in national office |
Ref document number: 2001916568 Country of ref document: EP |
|
WWW | Wipo information: withdrawn in national office |
Ref document number: 2001916568 Country of ref document: EP |