USES OF NEUREGULIN IN PREVENTING, TREATING OR DELAYING VENTRICULAR ARRHYTHMIA, AND COMPOSITION THEREOF

Abstract

Disclosed are uses of neuregulin in the preparation of medicines for preventing, treating or delaying ventricular arrhythmia of a human being, and pharmaceutical preparations that comprise the neuregulin and are used for preventing, treating or delaying the ventricular arrhythmia.

Claims

1. Use of NRG for the preparation of a medicament for preventing, treating or delaying ventricular arrhythmias in a mammal.

2. The use of claim 1, wherein the NRG is selected from NRG-1, NRG-2, NRG-3 or NRG-4.

3. The use of claim 1, wherein the NRG is NRG-1.

4. The use of claim 1, wherein the NRG comprises the amino acid sequence of SEQ ID NO: 2.

5. The use of claim 1, wherein the mammal is human.

6. The use of claim 1, wherein ventricular arrhythmias includes ventricular premature beat (VPB) and/or ventricular tachycardia (VT) and/or ventricular fibrillation (VF).

7. A pharmaceutical preparation for preventing, treating or delaying ventricular arrhythmias in a mammal, wherein the pharmaceutical preparation comprises an effective amount of NRG.

8. The pharmaceutical preparation of claim 7, wherein the NRG is selected from NRG-1,NRG-2,NRG-3 or NRG-4.

9. The pharmaceutical preparation of claim 7, wherein the NRG is NRG-1.

10. The pharmaceutical preparation of claim 7, wherein the NRG comprises the amino acid sequence of SEQ ID NO: 1.

11. The pharmaceutical preparation of claim 7,wherein the mammal is human.

12. The pharmaceutical preparation of claim 7, wherein ventricular arrhythmias includes ventricular premature beat (VPB) and/or ventricular tachycardia (VT) and/or ventricular fibrillation (VF).

13. A composition for preventing, treating or delaying ventricular arrhythmias in a mammal, wherein the composition comprises the pharmaceutical preparation of claim 7 and other drug(s) for treating ventricular arrhythmias.

14. The composition of claim 13, wherein other drug(s) for treating ventricular arrhythmias comprise a sodium channel blocker.

15. The composition of claim 13, wherein other drug(s) for treating ventricular arrhythmias comprise a β adrenergic receptor blocker.

16. The composition of claim 13, wherein ventricular arrhythmias includes ventricular premature beat (VPB) and/or ventricular tachycardia (VT) and/or ventricular fibrillation (VF).

17. A kit for preventing, treating or delaying ventricular arrhythmias in a mammal, wherein the kit comprises the pharmaceutical preparation of claim 6 and instructions on how to use the pharmaceutical preparation.

18. The kit of claim 17, wherein ventricular arrhythmias includes ventricular premature beat (VPB) and/or ventricular tachycardia (VT) and/or ventricular fibrillation (VF).

Description

BRIEF DESCRIPTION OF THE DRAWING

[0039] FIG. 1: Comparison of change of ventricular arrhythmias (VPB, VT, VF) incidence before administration relative to baseline between the two groups (p<0.001).

[0040] FIG. 2: Comparison of change of ventricular arrhythmias (VPB, VT, VF) incidence after administration relative to baseline between the two groups.

[0041] FIG. 3: Comparison of change of the sum of ventricular arrhythmias (VPB, VT, VF) incidence before/after administration relative to baseline between the two groups.

[0042] FIG. 4: Comparison of change of VPB incidence before administration relative to baseline between the two groups.

[0043] FIG. 5: Comparison of change of VPB incidence after administration relative to baseline between the two groups

[0044] FIG. 6: Comparison of change of the sum of VPB incidence before/after administration relative to baseline between the two groups

[0045] FIG. 7: Frequency distribution of QTc changes after each administration relative to baseline baseline to the subject.

[0046] The invention will be further illustrated by reference to the following examples. It should be understood that the examples are illustrative, but not limiting.

EXAMPLES

Example 1

A Randomized, Double-Blinded, Multi-Center, Placebo Controlled, Standard Treatment Based Study to Evaluate the Effect of Recombinant Human Neuregulin on the Survival of Patients with Chronic Heart Failure

[0047] To evaluate the efficacy of recombinant human neuregulin-1 for injection on chronic heart failure, a phase II, double-blinded, multi-center, placebo controlled, standard treatment based study was carried out in multiple clinical centers in China. A total of 351 patients with NYHA Class III or IV stable chronic heart failure were enrolled and randomized into two groups: placebo, or 0.6 μg/kg rhNRG-1. There were no significant variations in demographics or background therapies between the two groups. According to the schedule, patients were administered the drug for 10 consecutive days in the hospital first, and were allowed to be discharged from the hospital on the 11.sup.th day. Then they were administered the drug once a week as outpatient from the 3.sup.th week to the 25.sup.th week. Survival information was collected during the fifty-third week of the study. The 12 lead ECG was examined before and after each administration.

[0048] Investigational drug:

[0049] Specification: Neucardin™, the EGF-like domain of Neuregulin-1 β2 isoform is constituted of 61 amino acids, with the molecular weight of 7054 Dal (1 μg=0.14 nmol). 250 μg (5000 EU)/vial (1 μg=20 EU).

[0050] Placebo

[0051] Specification: excipients of Neucardin™ (250 μg/vial, without the active ingredient of recombinant human NRG-1 protein)

[0052] Dosage form: Powder injection.

[0053] Storage: in safe place, kept away from light, at 3-8° C.

[0054] Mode of administration: Intravenously drip or infusion.

[0055] Dosing regimen:

TABLE-US-00001 1-10 day 3-25 weeks Dosage 0.6 μg/kg/day 0.8 μg/kg/day rhNRG-1 or placebo rhNRG-1 or placebo Administration Intravenously drip Intravenously route infusion regimen 10 hours per day, for 10 minutes infusion consecutive 10 days each week

[0056] The clinical inclusion criteria: chronic heart failure patients with a NYHA classification of III or IV at the age of 18-65 were considered for enrollment if they had left ventricular ejection fraction (LVEF) of ≦40%, and stable clinical symptoms(including clinical symptoms, signs and standard therapy for heart failure has reached the target dose or maximum tolerated dose for more than 1 months).

[0057] Major exclusion criteria includes acute myocardial infarction, hypertrophic cardiomyopathy, constrictive pericarditis, significant valve disease or congenital heart disease, severe pulmonary hypertension, systolic blood pressure <90 mmHg or >160 mmHg, severe ventricular arrhythmia, cardiac surgery or a cerebrovascular event within the previous six months, claustrophobia or pregnant female subjects.

[0058] All patients provided written consent.

[0059] The study included three stages, namely, the screening period, the administration period and the follow-up period after drug withdrawal. The whole study period was 12 months (52 weeks). A total of 35 on-site visits and 2 telephone interviews was proceeded, and a number of indicators was observed during the study process (see Table 1).

TABLE-US-00002 TABLE 1 Study process and observational indicators screening Study Cycle and baseline 1 week 2 weeks 3 weeks 4 weeks Visit Frequency 1 2 3 4 5 6 7 8 9 10 11 12 13 Administration Frequency 1 2 3 4 5 6 7 8  9 10 11 12 Inclusion/Exclusion Criteria X Clinical History X Weight X Vital Signs X X X X X X X X X X X X X Drug Combination X X X X X X X X X X X X X Two Dimensional Echocardiogram X Heart Function Grade of NYHA X X Urine Pregnancy Test X X (childbearing Age) Routine Blood, Urine Test, Blood X X Biochemistry, CTnT Coagulation Function Test X X (APTT, PT) 12 Lead ECG X X X X X X X X X X X X X 6-min walk test X X NT-proBNP X X Anti-rhNRG-1β2 X X X Chest Radiography X Organization B ultrasonic (mammary gland, X liver and gall, spleen, pancreas, kidney, adrenal gland, pelvic cavity) Urine Volume for 24 hours X X X X X X X X X X X Administration of Study Drugs X X X X X X X X X X X X Dyspnea Assessment X X Life Quality Evaluation X X Adverse Event/Serious Adverse Event X X X X X X X X X X X X X Telephone Visits follow-up period Study Cycle 5-11 weeks 12 weeks 13-24 weeks 25 weeks 35 weeks 45 weeks 52 weeks Visit Frequency 14-20 21 22-33 34 35 Administration Frequency 13-19 20 21-32 33 Inclusion/Exclusion Criteria Clinical History Weight X X X Vital Signs X X X X X Drug Combination X X X X X Two Dimensional Echocardiogram X X X Heart Function Grade of NYHA X X X Urine Pregnancy Test X X X (childbearing Age) Routine Blood, Urine Test, Blood X X X Biochemistry, CTnT Coagulation Function Test X X X (APTT, PT) 12 Lead ECG X X X X X 6-min walk test X X X NT-proBNP X X X Anti-rhNRG-1β2 X X X Chest Radiography X Organization B ultrasonic (mammary gland, liver and gall, spleen, pancreas, kidney, adrenal gland, pelvic cavity) Urine Volume for 24 hours Administration of Study Drugs X X X X Dyspnea Assessment X X X Life Quality Evaluation X X X Adverse Event/Serious Adverse Event X X X X X X X Telephone Visits X X

[0060] Results and data analysis:

[0061] This study analyzed all ECG, described the abnormal change of ECG before and after each administration, analyzed the parameters changes of ECG before and after each administration, including heart rate, QRS interval, PR interval, RR interval, QT interval, QTc. The central tendency (mean, standard deviation, median) and classification analysis of QTc was carried out. Rank sum test was used in group comparison, and WILCOXON rank sum test was used for comparison between groups. Table 2 shows baseline of ECG diagnosis and parameters of all selected patients. The comparison of change of ventricular arrhythmias (VPB, VT, VF) incidence before each administration relative to baseline between the two groups was shown in Table 3 and FIG. 1. The comparison of change of ventricular arrhythmias (VPB, VT, VF) incidence after each administration relative to baseline between the two groups was shown in Table 4 and FIG. 2. The comparison of change of ventricular arrhythmias (VPB, VT, VF) incidence before/after each administration relative to baseline between the two groups was shown in Table 5 and FIG. 3. In addition, the VPB was further analyzed. The comparison of change of VPB incidence before each administration relative to baseline between the two groups was shown in Table 6 and FIG. 4. The comparison of VPB incidence change after each administration relative to baseline between the two groups was shown in Table 7 and FIG. 5. The comparison of VPB incidence change before/after each administration relative to baseline between the two groups was shown in Table 8 and FIG. 6. In addition, the frequency distribution analysis of QTc change of all patients after each administration was shown in FIG. 7.

TABLE-US-00003 TABLE 2 Comparison of ECG diagnosis and parameter between two groups of subjects at baseline Neucardin ™ Placebo Between Group Group groups ECG Normal ECG 5 (3.01%) 3 (1.80%) 0.340 Diag- Abnormal/ 16 (9.64%) 24 (14.37%) nosis NCS Abnormal/ 145 (87.35%) 140 (83.83%) CS Cardiac Sinus 128 (77.1) 129 (77.2) 0.976 Rhythm rhythm Arrhythmia 38 (22.9) 38 (22.8) VPB 19 (11.4%) 10 (6.0%) 0.077 VT 1 (0.6) 0 (0%) 0.498 VF 0 (0%) 0 (0%) Heart 77.2 ± 17.0 79.1 ± 16.1 0.2909 rate (n = 166) (n = 167) RR 829.16 ± 182.21 782.60 ± 168.90 0.0245 interval (n = 145) (n = 146) (ms) QT 405.05 ± 53.14  392.08 ± 45.16  0.0169 interval (n = 166) (n = 167) (ms) QTc 445.68 ± 45.45  433.61 ± 39.95  0.011 (ms) (n = 165) (n = 164) Note: NCS means non-clinical significance; CS means clinical significance

[0062] As shown in Table 2, the rate of abnormal ECGs (87.35% vs 83.83%) and rate of arrhythmias (22.9% vs 22.8%) between Neucardin™ and placebo group were comparable at baseline (p>0.05), however, RR interval (829.2±182.2 ms vs 782.6±168.9 ms) and QTc interval (445.7±45.5 ms vs 433.6±39.9 ms) were significantly longer for Neucardin™ group compared to placebo group (p=0.0245 and 0.011 respectively).

TABLE-US-00004 TABLE 3 The comparison of change of ventricular arrhythmia (ventricular premature beat, ventricular tachycardia, and ventricular fibrillation) incidence before administration relative to baseline in two groups of subjects (p < 0.0001) Neucardin ™ Placebo Group Group (N = 175) (N = 176) P value N 34 34 <.0001 Mean (SD) −0.168 (3.301) 3.417 (3.450) Median −0.333 3.175 Min, Max −7.16, 6.48 −2.42, 9.76

[0063] Table 3 shows the difference value between the incidence of ventricular arrhythmia before administration and the incidence of ventricular arrhythmia at baseline. N is the times of ECG diagnosis. Mean is mean value. Median is medium value. Min and Max are the minimum value and maximum value in these difference value, respectively.

TABLE-US-00005 TABLE 4 The comparison of ventricular arrhythmia (ventricular premature beat, ventricular tachycardia, and ventricular fibrillation) incidence after administration relative to baseline in two groups of subjects (p < 0.0001) Neucardin ™ Placebo Group Group (N = 175) (N = 176) P value N 34 34 0.1570 Mean (SD) 1.414 (3.764) 2.700 (3.640) Median 1.863 2.377 Min, Max −5.79, 7.67 −4.56, 11.39

[0064] Table 4 shows the difference value between the incidence of ventricular arrhythmia after administration and the incidence of ventricular arrhythmia at baseline. N is the times of ECG diagnosis. Mean is mean value. Median is medium value. Min and Max are the minimum value and maximum value in these difference value, respectively.

TABLE-US-00006 TABLE 5 The comparison of the sum of ventricular arrhythmia (ventricular premature beat, ventricular tachycardia, and ventricular fibrillation) incidence before and after administration relative to baseline in two groups of subjects (p < 0.0001) Neucardin ™ Placebo Group Group (N = 175) (N = 176) P value N 34 34 0.0007 Mean (SD) 1.246 (5.656) 6.117 (5.614) Median 1.368 5.265 Min, Max −10.88, 13.14 −5.66, 20.27

[0065] Table 5 shows the difference value between the sum of incidence of ventricular arrhythmia (ventricular premature beat, ventricular tachycardia, and ventricular fibrillation) in two groups of subjects before and after administration and the incidence of ventricular arrhythmia of baseline. N is the times of ECG diagnosis. Mean is mean value. Median is medium value. Min and Max are the minimum value and maximum value in these difference value, respectively.

TABLE-US-00007 TABLE 6 The comparison of change of VPB incidence before administration relative to baseline in two groups of subjects (p < 0.0001) Neucardin ™ Placebo Group Group (N = 175) (N = 176) P value N 34 34 <0.0001 Mean (SD) −1.158 (1.793) 2.303 (2.005) Median −1.100 1.800 Min, Max −4.60, 2.60 −1.90, 6.80

[0066] Table 6 shows the difference value between incidence of VPB in two groups of subjects before administration and incidence of VPB at the baseline. N is the times of ECG diagnosis. Mean is mean value. Median is medium value. Min and Max are the minimum value and maximum value in these difference value, respectively.

TABLE-US-00008 TABLE 7 The comparison of change of VPB incidence after administration relative to baseline in two groups of subjects (p < 0.0001) Neucardin ™ Placebo Group Group (N = 175) (N = 176) P value N 34 34 <0.001 Mean (SD) −0.145 (2.198) 1.952 (2.140) Median −0.400 1.600 Min, Max −4.40, 4.50 −1.70, 8.10

[0067] Table 7 shows the difference value between incidence of VPB in two groups of subjects after administration and baseline. N is the times of ECG diagnosis. Mean is mean value. Median is medium value. Min and Max are the minimum value and maximum value in these difference value, respectively.

TABLE-US-00009 TABLE 8 The comparison of the sum of VPB incidence before and after administration relative to baseline in two groups of subjects (p < 0.0001) Neucardin ™ Placebo Group Group (N = 175) (N = 176) P value N 34 34 <0.0001 Mean (SD) −1.303 (3.079) 4.255 (3.485) Median −1.500 3.700 Min, Max −7.80, 5.30 −0.80, 14.90

[0068] Table 8 shows the difference value between the sum of incidence of VPB in two groups of subjects before and after administration and incidence of VPB at baseline. N is the times of ECG diagnosis. Mean is mean value. Median is medium value. Min and Max are the minimum value and maximum value in these difference value, respectively.

[0069] Conclusions:

[0070] The incidence of arrhythmias at baseline were 22.9% and 22.8% in the Neucardin™ and placebo group respectively, including VPB incidence of 11.4% and 6.0%, VT incidence of 0.6% and 0%, VF incidence of 0% and 0% in the Neucardin™ group and placebo group, respectively. There were no significant differences for VPB, VT and VF incidence between the two groups.(see Table 2)

[0071] The data shown in Tables 3, 4, and 5 indicate that among the ECG tests throughout the study (before/after study drug administrations, 68 times in total), there was a very significant decreasing trend of incidence of ventricular arrhythmias in the Neucardin™ group compared with the placebo group before and after administration (p<0.0001).The results were shown in FIGS. 1, 2, and 3.

[0072] In addition, the ventricular premature beat was further analyzed. The change of incidence of premature ventricular beat before/after administration relative to baseline, as shown in table 6, 7 and 8, indicated that there is a very significant decreasing trend of the incidence of ventricular premature beat in the Neucardin™ group compared with the placebo group (p<0.0001). The results were shown in FIGS. 4, 5, and 6.

[0073] Although the RR interval, QT interval and QTc interval were longer in Neucardin™ group than in the placebo group at baseline, the QTc intervals were not progressively prolonged after Neucardin™ administration compared to baseline, instead, the majority showed a tendency of significant shortening. Analysis of the QTc change frequency distribution after Neucardin™ administration indicated that the QTc intervals of Neucardin™ group generally showed a shortening trend compared with placebo group (FIG. 7).

[0074] Comprehensive analysis of the ECG diagnostics and parameters indicates that although 87.35% subjects in Neucardin™ group were with ECG abnormalities at baseline which includes atrial fibrillation (AF), atrioventricular block, premature ventricular contraction, and ST-T changes, these ECG abnormalities were not found to be progressively increased or worsened after Neucardin™ administration. The results show that the QTc of Neucardin™ group of subjects showed a general shortening trend compared with placebo group, and the change of incidence of ventricular arrhythmia and/or ventricular premature beat significantly reduced compared with the placebo group.

[0075] The examples listed above do not limit the protection scope of the invention. Without departure from the purposes and scope of the present invention, those of ordinary skill in the art may adjust and change the present invention. Therefore, the protection scope of the invention shall be defined by the claims, rather than by specific examples.