Use of GHRP-6 as late cardioprotective and cardiac restoration medicament

Abstract

The present invention is related to the use of the growth hormone releasing peptide-6 (GHRP-6) for the manufacture of a late cardioprotective and cardiac restoration medicament. Said late cardioprotective and cardiac restoration medicament comprises GHRP-6 and a pharmaceutically acceptable excipient or vehicle. It also provides a method for the treatment of a disease that involves a low cardiac output, wherein a therapeutically effective amount of a late cardioprotective and cardiac restoration medicament comprising GHRP-6 is administered to a subject in need. Said medicament allows treating in a late manner, even days later, the myocardium that suffered episodes of stunning, hibernation, ischemia and its consequences thereof.

Claims

1. A method for the treatment of a disease that involves a low cardiac output, said method comprising administering to a human in need thereof a therapeutically effective amount of a late cardioprotective and cardiac restoration medicament comprising the growth hormone releasing peptide-6 (GHRP-6), wherein the medicament is first administered at least 12 hours after an episode of ischemia is established.

2. The method according to claim 1 wherein the medicament is administered to patients affected by a pathology selected from the group consisting of acute myocardial infarction (AMI) with elevation or depression of the ST segment, acute coronary syndrome, acute or chronic ischemic myocardial disease, heart failure, episodes of ischemia/reperfusion of the heart muscle, to rescue ventricular mass from stunning or hibernation states, low-cardiac output syndrome and cardiogenic shock.

3. The method according to claim 1 wherein the medicament is administered to patients awaiting a heart transplant to correct and optimize the systemic homeostasis and general state of health.

4. The method according to claim 1 wherein the medicament is administered by central intravenous route, peripheral intravenous route or within the coronary tree.

5. The method according to claim 4 wherein the medicament is administered as part of an endoluminal dilation procedure.

6. The method according to claim 1 wherein the GHRP-6 is administered at 25-200 μg/kg of a patient's body weight as a bolus.

7. The method according to claim 1 wherein the medicament is first administered between 13 and 96 hours after an episode of ischemia is established.

Description

DESCRIPTION OF THE INVENTION

(1) This invention contributes to solve the aforementioned problem, disclosing the use of the GHRP-6 in the manufacture of a late cardioprotective and cardiac restoration medicine. For the first time, a pharmacological agent able to restore cell homeostasis and function of cardiac tissue, for long hours or even days, of an established ischemic process, regardless of the current narrow time window that impose the biology of the system itself, and able to restore the cellular metabolism has been identified. In this way, the window of opportunity for treatment and, therefore, the possibility for therapeutic success is extended.

(2) The invention is based on the use of GHRP-6 as an active pharmaceutical ingredient capable of restoring cardiomyocytes function, in late scenarios of acute or chronic, mild or severe, acute or terminal ventricular failure. The treatment ensures that a set of molecular processes that involve the salvation route known as RISK (Reperfusion Injury Salvage Kinase) get activated. In addition, it restores the function of ionic pumps, and reduces inflammation, wall stress, and the formation of reactive oxygen species.

(3) For the purposes of the invention, the term “cardioprotective medicine” refers to a drug capable of rescuing cardiomyocytes from cell death, in scenarios that otherwise were lethal, being able to maintain energy homeostasis, the stability of ionic pumps, to guarantee conduction and rhythm, as well as to maintain the contractile function of such types of cells.

(4) The term “cardiac restoration medicine”, as used in this invention, refers to a medicament capable of reversing the deterioration of myocardial mechanical function, without increasing heart rate, oxygen consumption, cardiac output, or at the cost of reducing diastole time.

(5) GHRP-6, as an active pharmaceutical ingredient is applied with therapeutic purposes, even belatedly in all said subjects with myocardial damage or dysfunction due to hibernation or ventricular remodeling. It can also be applied to subjects undergoing coronary revascularization interventions, in which the oxygenation of one or several organs is necessarily compromised, due to total or partial ischemia, and then reperfusion is performed. It is also applicable in the control and prevention of the consequences of thromboembolic phenomena. Even more relevant, the treatment is applicable to those who are revascularized and who cannot recover the contractile function of the ventricle and progress to systolic and diastolic dysfunctions.

(6) Therefore, in an embodiment of the invention, the cardioprotective and cardiac restoration medicament that comprises GHRP-6 is administered to patients affected by a pathology selected from the group comprising: AMI with elevation or depression of ST segment, acute coronary syndrome, acute or chronic myocardial ischemic disease, heart failure and ischemia/reperfusion episodes of the cardiac muscle, to rescue ventricular mass from stunning or hibernation, low cardiac output syndrome and cardiogenic shock.

(7) The use of GHRP-6 may be the only pharmacological option to control stunning and hibernation, which can ultimately lead to heart failure syndrome and to death. The compassionate use of GHRP-6 has shown to be useful in critically ill patients with Grade IV Heart Failure, and in others already waiting for an appropriate donor for heart transplantation. It presupposes the rupture of two paradigms: (1) Rupture of the golden rule that “time is myocardium”, in the way that the time elapsed in the implantation of the damage is already irrelevant as opposed to the efficacy of the medication; (2) Stunning, hibernation, low output and severe heart failure are already pharmacologically manageable. These are clear advantages of this invention. Therefore, in another embodiment, the late cardioprotective and cardiac restoration medicament of the invention is administered to patients awaiting a heart transplant, to correct and optimize their systemic homeostasis and health status, in general.

(8) In one embodiment, the late cardioprotective and cardiac restoration medicament comprising GHRP-6 is administered by peripheral intravenous, central or within the coronary tree. In a particular embodiment, said medicament is administered as part of the endoluminal dilatation procedure. The GHRP-6 can be administered by repeated injections or by continuous infusion systems. It includes intravenous and arterial administrations, including instillation within the coronary tree before or during reperfusion by balloon or “stent”. The versatility of the mechanisms of action of this agent justifies its administration to a wide range of patients who have suffered coronary episodes, any kind of cardiomyopathies, and regardless of the time of initiation of treatment and its extension. The merit of the treatment is to restore the function of the impaired heart, in an acute or stunned condition, and even beyond the so-called critical time or point of no return.

(9) In another scenario, the treatment can be applied therapeutically to graft recipients, to allow a proper conditioning of the host organism and the transplanted organ. Patients with grade IV functional class and with transplant criteria have been treated, resulting in a rise of the left ventricle mechanical function and they have reached the transplant under conditions of better systemic tissue oxygenation. It can be administered repeatedly, after the transplant is performed, and treatment can begin in the operating room, once the circulation between the recipient and the grafted organ has been established.

(10) In an embodiment of the invention, the medicament comprising GHRP-6 is administered in such a way that the patient receives between 25 and 200 μg of peptide/kg of body weight, in bolus form. The restorative effect of this treatment on the ischemic heart makes an option the concomitant administration of any antioxidant agent, of iron chelating compounds, as well as of enzymes involved in the anti-oxidant defense.

(11) In another aspect, the invention provides a late cardioprotective and cardiac restoration medicament comprising the GHRP-6 and a pharmaceutically acceptable vehicle or excipient. Due to its effect, late cardioprotective medicament comprising GHRP-6 can be administered 12 or more hours after the implantation of an episode of ischemia. In a particular embodiment, said medicament is administered between 13 and 96 hours after the implantation of an episode of ischemia.

(12) The late cardioprotective and cardiac restoration medicament of present invention may be concomitantly administered with thrombolytic or fibrinolytic agents, referred to as plasminogen activators, and in the event of reperfusion induced by mechanical methods, including angioplasty. Said medicament could be applied along, previously or subsequently to pre- and post-conditioning maneuvers.

(13) It is recommended to use concentrations ranging from 25 to 400 μg/ml, in vehicles such as physiological saline (0.9% sodium chloride); Ringer Lactate solution; plasma; albumin; and dextrose; isotonics or their mixtures. The routes of administration may be deep or peripheral intravenous, intraarterial or intraperitoneal. The administration of the drug of the invention can be repeated twice to three times a day.

(14) In the invention, GHRP-6 is administered, as late cardioprotective and cardiac restoration medicine, to patients with episodes of ischemia/reperfusion of the heart muscle, to correct the central and peripheral hemodynamics, stabilize the central venous pressure and the mean arterial pressure, as well as to eliminate vasoplegia and vascular resistance. In addition, it is administered to patients with lactic acidosis derived from states of low cardiac output, hypovolemia, ventilatory failure, increased preload.

(15) In another aspect, the invention discloses a method for the treatment of a disease that involves a low cardiac output wherein a therapeutically effective amount of a late cardioprotective and cardiac restoration medicament comprising GHRP-6 is administered to a subject in need thereof.

(16) In an embodiment of the invention, the method is applied to patients suffering a pathology selected from the group consisting of AMI with elevation or depression of ST segment, acute coronary syndrome, acute or chronic ischemic myocardial disease, heart failure, episodes of ischemia/reperfusion of the heart muscle, to rescue the ventricular mass from states of stunning or hibernation, low-cardiac output syndrome and cardiogenic shock. In another embodiment, the method is applied to patients awaiting a heart transplant, to correct and optimize their systemic homeostasis and health status, in general.

EXAMPLES

Example 1. Demonstration of the Effect of GHRP-6 on the Extension of the Window of Therapeutic Opportunity for the Rescue of Myocardial Mass Under Coronary Occlusion. Experimental Model With and Without Reperfusion

(17) The pharmacological manipulation, to reduce the size of the infarction, has not yielded the expected results, despite 45 years of incessant research. The sudden character of the ischemic episode contributes to this and, especially, the short time available to rescue ventricular mass from death when ischemia has been implanted.

(18) The objective of the study was to evaluate, and to compare, the therapeutic impact of the late administration of GHRP-6 on functional parameters of the left ventricle, regarding the effect of the peptide acutely administered. Another objective was to assess the effect of late treatment in a group of animals in which the reperfusion was not performed.

(19) The implementation of the AMI was performed in female pigs of the Yorkshire breed, with body weight of 22-27 kg. The anesthesia was induced with propofol, midazolam and pancuronium, by intravenous route. Under general anesthesia, obtained with a mixture of isofluorane/oxygen and nitrous oxide the ischemia was provoked. It was induced by a thoracotomy between the fifth and sixth intercostal spaces, until reaching the left circumflex artery, which was occluded by a 30 mm arterial clamping for one hour. Immediately, the record of the electrocardiogram and the heart rate was taken, every 20 seconds. At the end of 60 minutes of occlusion, the clamping was released, and so began the reperfusion phase, to complete the 10 days after ischemia. For the experimental group without reperfusion, that simulates the state called as non-reflux, a permanent ligation of the circumflex artery, with surgical thread, was performed. Both procedures, clamping and permanent ligation, induce an AMI with a ST Segment elevation. The recording of heart rate and electrocardiogram continued until the superficial suture of the thorax. The animals were sacrificed, by exsanguination, on the tenth day after the ischemia episode was induced. Under an anesthetic overdose, the determination of the area of the AMI was made, by means of the calculation of the area at risk with Evans blue.

(20) For the echocardiographic evaluation, the pigs were previously sedated with a mixture of propofol/midazolam. A Kontron ultrasound system, Sigma 1 AC Cardio model, equipped with 3.5-7.5 MHz mechanical-cardiac sector transducers was used.

(21) The parameters evaluated were the following: Left Ventricle Ejection Fraction (LVEF), estimated by the Teich method. Left Ventricle Tele-Systolic and Tele-diastolic Pressures. Infarction Area, estimated on the area at risk (%).

(22) The study design, in terms of the experimental groups used, is summarized in Table 1. Seven animals per group were used. In all groups, the animals received treatment every 12 hours, for 10 days after ischemia. The administrations of GHRP-6 or placebo were performed through a central catheter. In the case of the groups that received the peptide, it was diluted in 10 ml of physiological saline. The essence of the study was to evaluate the influence of the time of initiation of treatment on functional parameters and size of the ventricular not viable territory.

(23) TABLE-US-00001 TABLE 1 Experimental design. Group Onset of treatment 1. Physiological saline Immediately at the beginning of reperfusion (Placebo) 2. GHRP-6 (400 μg/kg) Immediately at the beginning of reperfusion 3. GHRP-6 (400 μg/kg) 72 hours after the start of reperfusion 4. GHRP-6 (400 μg/kg) 5 days after the start of reperfusion 5. No reperfusion One hour after the artery was permanently (GHRP-6 400 μg/kg) clamped and after the occurrence of an elevation in the ST segment.

(24) The response to treatment with GHRP-6, administered at different times after reperfusion, is reflected in Table 2. The evaluation of the effect of the treatment on the control of the infarct size as well as variables describing the function and state of the left ventricle are included. The results indicate that, even if the treatment is started 72 hours after restoration of blood flow (reperfusion), GHRP-6 is able to significantly reduce the size of myocardial infarction by 13%, approximately, in comparison to placebo. There were no significant statistically differences between the effect of intervention with GHRP-6 in parallel to reperfusion and the one achieved when the treatment started 72 hours later. Although more discreetly, the latest treatment, initiated on the 5th day after ischemia, still rescue from death an 8% of the ventricular territory corresponding to the area at risk. The result of opening a window of therapeutic opportunity in the non-reperfused or non-reflux group was even more surprising.

(25) TABLE-US-00002 TABLE 2 Response variables to treatment. LV* LV telediastolic telesystolic Experimental Infarction pressure pressure Group Area (%) *LVEF (%) (mmHg) (mmHg) Basal Not 81 ± 2.8 3.3 ± 1.7 89 ± 2.3 conditions applicable Group 1 48.7 ± 2.6 38 ± 3.6 9.5 ± 2.2 68 ± 4.2 Group 2 31.6 ± 5.2 80 ± 2.5 4.6 ± 1.1 80 ± 2.2 Group 3 35.4 ± 6.4 77 ± 4.1 4.4 ± 1.6 81 ± 3.0 Group 4 37.0 ± 4.3 60 ± 6.6 5.2 ± 0.8 77 ± 3.1 Group 5 36.1 ± 2.8 72 ± 5.5 5.1 ± 1.9 80 ± 2.6 **LV: Left Ventricle

(26) Even with no rescue effect associated to the phenomenon of reperfusion, the treatment with GHRP-6 reduced by 12% the area of ventricular death. Corresponding to the effect of anti-necrogenic rescue still late, it is notorious the impact of the treatment on the preservation of LVEF. The AMI caused a 43% decrease in LVEF in the placebo group, 10 days after the episode. However, in the most astringent context, associated to the start of treatment with GHRP-6 on the fifth day, the deterioration of ventricular function only declines by 21%, compared to the study in baseline physiological conditions. In other words, 22% of the LVEF is rescued in the group that began to be medicated 5 days after the ischemic event. Similarly, the effect of treatment with GHRP-6 in the non-reflux group is also surprising. Even without reperfusion, the treatment prevents cell death and guarantees the function of pump of the left ventricle with only 9% difference with the data derived from baseline conditions.

(27) On the other hand, the GHRP-6 supports the parameters of telesystolic and telediastolic pressures in a way similar to those determined in baseline physiological conditions. There is a significant reduction in diastolic pressure in all the groups that received the GHRP-6, regardless of when the treatment started, when compared to the placebo group. In concordance, the treatment manages to maintain the intra-cavity pressure at the end of the systole much higher than that determined for the placebo group. In other words, the treatment prevents the acute ventricular pump failure or the low cardiac output syndrome.

(28) It can be concluded that the treatment with GHRP-6 showed a surprising and unusual ability to expand the window of therapeutic opportunity in episodes of ischemia and heart reperfusion, such as those associated to AMI. In addition, the treatment with GHRP-6 showed a surprising and unusual ability to restore cardiac cell homeostasis, as well as its vital functions, in the most pathogenic of the scenarios that is in the non-reperfusion states. It establishes a new property, the ability to expand the window of functional opportunity. Also, it was evidenced that the treatment aborts the phenomenon of ventricular stunning associated to ischemia. Additionally, an AMI with five days of evolution appears, for the first time, as a scenario pharmacologically manageable with GHRP-6, restoring functional homeostasis in a proper way.

(29) As infarct progresses as “a wavefront” from the subendocardium to the epicardium, it is possible to abort this wave progress toward death, by widening the window of therapeutic opportunity for reperfusion or other required pharmacological interventions. It is shown, for the first time, that is possible to establish homeostasis of the heart muscle even when days have elapsed since the onset of the ischemic episode. This finding constitutes an unprecedented milestone in medicine.

Example 2. Extension of the Therapeutic Opportunity Window for the Functional Restoration of Myocardium in Patients with Acute Infarction Treated with GHRP-6

(30) Elements of clinical response in patients who suffered a myocardial infarction with elevation of the ST segment, treated with GHRP-6 in an acute or a delayed way are shown here below.

(31) First cohort. Comprising 19 patients over 30 years old, of both sexes, with a diagnosis of AMI with ST segment elevation, all with occlusion of the anterior descending artery, first episode, who arrived at a cardiology department before 12 hours of implantation of the precordial pain. The average door-ball time was 6 hours. All patients received primary percutaneous transluminal angioplasty, and immediately after the culprit artery flow was restored they received treatment with GHRP-6. In some patients, the treatment was performed by intra-coronary flushing, after the flow was restored. Residual lesions occupied less than 25% of the initial occlusion area, so it was considered as a TIMI 3, according to the Coronary Risk Score known as TIMI. In other cases, the first administration was performed by a central-venous route, just at the time of reperfusion, in the hemodynamic room. GHRP-6 was administered at a dose of 100 μg/kg of body weight, twice a day, for 7 days, time spent in the coronary intensive care hospital room. Clinical, echocardiographic and scintigraphy perfusion imaging evaluation were performed 30 days after the episode of ischemia.

(32) Second cohort. Comprising 13 patients over 30 years old, of both sexes, with a diagnosis of AMI with elevated ST segment, who received percutaneous transluminal angioplasty before 12 hours of implantation of precordial pain. All these patients evolved in an unfavorable way, and with uncertain prognosis between 24 and 96 hours after a successful endoluminal reperfusion, according to the imaging criteria all were considered as TIMI grade 3. Hypothetically, this apparent success of reperfusion associated with a ventricular dysfunction, reminiscent of myocardial stunning, may be due to two phenomena, the non-restoration of circulation at the microcapillary system level, suggesting the persistence of ischemia/tissue hypoxia; or the existence of true stunning considering a TIMI 3, with poor ventricular contractility.

(33) These patients had a galloping evolution, from low-cardiac output syndrome to cardiogenic shock. This led to the decision of establishing an alternative treatment with GHRP-6, to try to reduce dyskinesia of the ventricular musculature, to attenuate the process of presumed stunning, to dampen hypoxia, and to rescue from death a territory of ischemic ventricular mass. The patients who implanted a reinfarction received rescue angioplasty. One of these patients died during the procedure. None of the patients met criteria for the coronary revascularization practice. A summary of the characteristics of the patients in that second cohort is shown in Table 3. In these cases, the first administration of the peptide was carried out at least after 48 hours of torpid evolution after a TIMI 3. During the treatment time, the GHRP-6 was administered intravenously, every 12 hours.

(34) TABLE-US-00003 TABLE 3 Demographic characterization and evolution of the patients in the second cohort. Patient Sex/Age PTA Time Evolution between 3 and 7 days RGD F/65  6 hours LCOS AFP M/49  4 hours CS YRH M/71 11 hours CS PPL M/62  7 hours LCOS ICR F/70 10 hours CS + SIRS JIM F/66 11 hours LCOS + CS AMG M/72  9 hours CS CVP M/59 10 hours CS + SIRS CSC M/51  4 hours Reinfarction. Patient died AEA F/66  8 hours LCOS MPS M/65 10 hours CS NSS F/60  6 hours Reinfarction at 24 hours. YMM M/55  6 hours Reinfarction on the third day. PTA: Percutaneous Transluminal Angioplasty, LCOS: Low Cardiac Output Syndrome, CS: Cardiogenic Shock, SIRS: Systemic Inflammatory Response Syndrome.
Results of the Study

(35) The patients of the first cohort, who received the GHRP-6 after a successful percutaneous transluminal angioplasty with reperfusion considered as TIMI 3, exhibited a favorable evolution, without further important complications. The initial and evolutionary echocardiographic parameters are shown in Table 4 (useful data derived from 15 patients). It is worth noting the response to treatment with GHRP-6, compared to the natural evolution of the disease, if the standard values taken as a reference in international literature are considered (Pedro R. Moreno and Juan H. del Portillo. Rev Colomb. Cardiol. 2016; 23 (6): 500-507; 2017 ESC Guidelines. European Heart Journal 2018; 39: 119-177).

(36) TABLE-US-00004 TABLE 4 Parameters measured by echocardiography performed to patients with AMI with ST segment elevation receiving GHRP-6 in an early manner. Telesystolic Telediastolic Time of SPWT SST volume volumen LVEF SSF Scanning (mm) (mm) (ml) (ml) % % Start of 10.1 ± 2   10.6 ± 3, 1 77 ± 12 81 ± 14  38 ± 5.9  27 ± 2.6 treatment. Minutes after the PTA. 30 days 11.6 ± 0.7  10.3 ± 1.6 65 ± 23 87 ± 11  51 ± 8.6  35 ± 6.5 after PTA 90 days 11.3 ± 1.8  10.7 ± 1   49 ± 8  111 ± 16   57 ± 6.6  42 ± 9.6 after PTA PTA: Percutaneous Transluminal Angioplasty; SPWT: Systolic Posterior Wall Thickness. Reference value (9.8-11.2 mm); SST: Systolic Septum Thickness, Reference value (8.6-11.7 mm), LVEF. Reference value (40-85%); SSF: Septum Shortening Fraction. Reference value (28-42%), Telesystolic Volume. Reference value (12-60 ml), Telediastolic Volume. Reference value (120-140 ml).

(37) It was found that patients treated with GHRP-6 showed progressive improvement over time. In all these cases, a physiological functional restitution was achieved, which is described below: There were no variations in morphological constants such as SPWT and SST. There was a progressive reduction of telesystolic volume, implying an improvement in pump function of the left ventricle, improving cardiac function and conservation of functioning cavities and the response of the wall, expressed normal levels of afterload. It was observed a progressive and stable increase of telediastolic volume, suggesting an improvement in diastolic dysfunction, increasing in extension of the left ventricle walls. In other words, better ventricular relaxation, with the subsequent increase in afterload. Finally, two nearby parameters, LVEF and AF, showed a steady increase, reaching normal levels at 90 days. This means a total correction of the segmental and global kinetics of the left ventricle function.

(38) Regarding the patients of the second cohort, who received treatment with GHRP-6 administered in a late manner, Table 5 shows the outcome, as well as the echocardiographic characterization, performed at 90 days of evolution.

(39) TABLE-US-00005 TABLE 5 Outcome and evolutionary results of the second cohort. Late treatment with GHRP-6. GHRP-6 Echocardiographic Patient intervention Dose Outcome characterization RGD Day 3 to 9 100 Elevation of LVEF Non-contractile area μg/kg after 72 hours of of 12% of the left treatment. Exit of ventricle. the low-cardiac LVEF of 63%. output syndrome. AFP Day 3 to 9 100 Elevation of LVEF. Non-contractile area μg/kg Increase in diuresis. of 21% of the left ventricle. LVEF of 48%. YRH Day 4 to 10 100 Elevation of LVEF. Non-contractile area μg/kg Disconnect the of 20% of the left mechanical ventricle. ventilation on the LVEF of 50%. fifth day. PPL Day 2 to 8 100 Elevation of LVEF Non-contractile area μg/kg after 72 hours of of 3% of the left treatment. ventricle. LVEF of 50%. ICR Day 3 to 9 200 Elevation of LVEF Non-contractile area μg/kg after 72 hours of of 17% of the left treatment. ventricle. Hemodynamic LVEF of 47%. stability. JIM Day 4 to 10 200 Elevation of LVEF Non-contractile area μg/kg after 72 hours of of 19% of the left treatment. ventricle. Hemodynamic LVEF of 51%. stability. Increase in diuresis AMG Day 4 to 10 100 Elevation of LVEF Non-contractile area μg/kg after 48 hours of of 13% of the left treatment. ventricle. Hemodynamic LVEF of 52%. stability CVP Day 4 to 10 200 Elevation of the Non-contractile area μg/kg LVEF at 48 hours. of 23% of the left Hemodynamic ventricle. stability LVEF of 46%. and diuresis on the fifth day post- treatment. AEA Day 3 to 9 100 Elevation of 7% non-contractile μg/kg the LVEF area of the left 48 hours after the ventricle. GHRP-6 started. LVEF of 56%. Hemodynamic stability and diuresis achieved on the fifth day of treatment. MPS Day 4 to 10 100 Elevation of LVEF Non-contractile area μg/kg after 48 hours of of 23% of the left treatment with ventricle. GHRP-6. LVEF of 45%. Hemodynamic stability with suspension of dobutamine and epinephrine on the fourth day of treatment.

(40) Patient CSC died during the angioplasty rescue procedure. Once the treatment with peptide was started, the suspension of vasopressor amines was achieved in 8 out of 10 patients evaluated, in an interval that was close to 96 hours. Of the 12 surviving patients, 10 were evaluated by echocardiography at the third month after AMI. It was surprising the response to the late treatment with GHRP-6 in all these patients. The major medical findings in these 10 patients demonstrate that treatment with said peptide favored survival, by widening the management and pharmacological intervention window in a comprehensive way. The non-existence in this technical field of a similar treatment brings an unquestionable novelty to this method of treatment.

(41) Concerning the 13 critical patients, 12 of them survived. At commencement of the study, all these patients showed very unfavorable evolution, with life threatening, despite a successful reperfusion maneuver, according the monitoring imaging criteria. The evolution towards low cardiac output syndrome, and cardiogenic shock, indicates that the pump function of left ventricle was not recovered, or that a severe and sustained ventricular stunning pattern was implanted. This implies the persistence of distal, partial, or even normoxia ischemia but with sarcomeric dysfunction. The treatment with GHRP-6 helped to stabilize the central and peripheral hemodynamics as well as the ejector function and cardiac output, reducing the supra-ventricular tachycardia, and increasing the ejection fraction. In addition, the treatment with GHRP-6 restored the glomerular filtrate, and diastolic function, reducing preload and venous pressures.

(42) Unexpectedly, the echocardiographic study conducted 90 days after the AMI with ST segment elevation shows dramatically favorable values of ejection fraction, of global contractility of the left ventricle and the septum. As it is shown in Table 6, 90 days after treatment with GHRP-6, the group treated in a late manner exhibited a LVEF close to 51%, which is unexpectedly favorable. Accordingly, the treatment with GHRP-6 favored the cell restoration, even when treatment began lately. All patients were treated with GHRP-6 at least 48 hours after developed complicated clinical conditions of hemodynamic instability and extra cardiac-dysfunctions were established. Therefore, it is demonstrated that the treatment with GHRP-6 significantly and unexpectedly expands the window of therapeutic opportunity for the restoration of cardiovascular functions in infarcted patients with life-threatening complications.

(43) TABLE-US-00006 TABLE 6 LVEF values determined 90 days after treatment with GHRP-6. Cohort LVEF %. Mean ± SD Cohort 1. Acute treatment 57.0 ± 6.61 Cohort 2. Late treatment 50.8 ± 5.34

Example 3. Clinical Demonstration of the Extension of the Therapeutic Opportunity Window in Patients with Grade IV Heart Failure Included in the Transplant Pool and Treated with GHRP-6

(44) This is a cohort of 7 patients, who received the compassionate treatment with GHRP-6, under informed consent. All patients were in advanced heart failure, evaluated as functional class grade IV, according to the scale of the New York Heart Association. These patients were hospitalized in specialized cardiology services of two hospitals, and the ischemic heart disease, the idiopathic dilated cardiomyopathy, the heart valve disease, and the postpartum dilated cardiomyopathy were considered as the underlying condition for heart failure. All patients evaluated had a significant limitation in functional capacity, were permanently bedridden at 45° Fowler position, with advanced cardiac dysfunction, and LVEF values below 25%. They had minimal effort dyspnea, inability to adequately fulfill the 6 minutes' walk test, and instability in fluid balance and renal function, despite the diet control and the treatment with diuretics, the restriction of salt and the monitoring body weight.

(45) All patients received between 50 and 200 μg of GHRP-6/kg body weight, two or three times a day, according to medical criteria, by intravenous route. All treatments were performed while the patients were awaiting for heart transplantation. The treatment period with GHRP-6 was between 25 days and 30 days per cycle.

(46) A cycle of treatment with GHRP-6 was performed every three months. The maximum was three cycles. All patients were maintained with previous medical treatments, based on digital, diuretics, renin-angiotensin-aldosterone axis inhibitors (neurohormones) and pressor amines when required. Table 7 summarizes initial and final (after treatment) values for each parameter.

(47) TABLE-US-00007 TABLE 7 Comparison of parameters evaluated before and after treatment in grade IV heart failure patients. Cardiac Oxygen 6 minute's Systolic output consumption Patient test (min)* volume* (L/m)* (ml/kg/min)* LVEF %* AMJ 2/23 25/47  2.1/2.89 4.2/6.6 22/31 BLP 41/201 27/51 1.98/3.2  3.63/9.5  18/26 BCC 38/258 29/50 3.2/4.3 4.12/7.7  20/28 IBP 27/198 30/54 2.7/3.8 3.91/7   19/30 NFG 39/179 29/60 1.89/2.9  3.05/6.4  21/29 MGT 29/221 31/56 2.17/3.2  4.1/7.2 20/23 JSA 35/175 35/49 2.41/3.08 3.87/7.6  21/25 *Before treatment values/after treatment values

(48) Compassionate treatments, performed on patients incorporated into the transplant pool, were applied to individuals in cardiovascular catastrophe with Grade IV heart failure. This presumes the harshest scenario for a pharmacological response, given the unfavorable prognosis involved in the process. The treatment was applied to patients with slight multiple organ dysfunction, due to its high degree of hemodynamic mismatch and systemic hypoxia.

(49) Despite this marked morbidity and co-morbidity, an unusual response was observed. As it is shown in Table 7, the treatment was able to restore cardiac, hemodynamic, ventilatory and clinical constants. The treatment showed that, regardless of the stage of cardiovascular and systemic deterioration, it is possible to correct homeostasis, and therefore vital parameters and functions. This evidence has no precedent in medical practice.