WO2014086819A1

WO2014086819A1 - Methods and pharmaceutical composition for the treatment and prevention of cardiac arrhythmias

Info

Publication number: WO2014086819A1
Application number: PCT/EP2013/075463
Authority: WO
Inventors: Jean-Yves Le Guennec; Thierry Durand; Camille OGER; Jean-Marie GALANO; Jérôme THIREAU; Valérie BULTEL-PONCE; Alexandre Guy; Jérôme ROY
Original assignee: INSERM (Institut National de la Santé et de la Recherche Médicale); Centre National De La Recherche Scientifique (Cnrs); Université De Montpellier 1; Université Montpellier 2 Sciences Et Techniques
Priority date: 2012-12-05
Filing date: 2013-12-04
Publication date: 2014-06-12

Abstract

The present invention relates to a F₄-neuroprostane (F₄-NeuroP) for use in the treatment and prevention of cardiac arrhythmias in a subject in need thereof.

Description

METHODS AND PHARMACEUTICAL COMPOSITION FOR THE TREATMENT AND PREVENTION OF CARDIAC ARRHYTHMIAS

FIELD OF THE INVENTION:

The present invention relates to methods and pharmaceutical compositions for the treatment and prevention of cardiac arrhythmias.

BACKGROUND OF THE INVENTION:

Cardiac arrhythmia is a term for any of a large and heterogeneous group of conditions in which there is abnormal electrical activity in the heart. The heart beat (pulse) can be too fast or too slow and can be regular or irregular. Cardiac arrhythmias present a significant health problem. Indeed some arrhythmias are life -threatening medical emergencies that can result in cardiac arrest and sudden death. Cardiopulmonary bypass; injury to the conduction system during surgery; and metabolic and electrolyte abnormalities, especially hypokalemia and hypomagnesemia, contribute to the increased incidence of postoperative arrhythmias. Stress of the surgery with enhanced sympathetic tone and use of ionotropic support are added factors.

Some fishes are rich in particular fatty acids, the omega-3 poly-unsaturated fatty acids (omega-3 PUFA). The main PUFAs are eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) which are highly peroxidable. The effects of omega-3 PUFA on cardiac function are still debated, notably because of the lack of information on mechanisms. For example, it is not really known which the active lipid is: the PUFA or one of their oxygenated metabolites. A prospective study on a large number of patients showed that the most marked effect of PUFAs is a reduction of sudden cardiac death in the months following a cardiac infarction¹. This benefit has been explained by a reduction of arrhythmias and of systolic cardiac failure, in parallel with other cardioprotective effects of omega-3 PUFA ². Experiments on single cardiac cells have shown that EPA and DHA can modulate the activity of ion channels, the transmembrane proteins responsible for the electrical activity of the heart ³, which allowed establishing a mechanistic basis for the antiarrythmic effects of omega-3 PUFA. However, concomitantly, it was also shown that the effects of DHA on some rat cardiac ion channels are correlated with the oxidation of the fatty acid and not with the fatty acid itself ⁴. Because intravenous inj ection of an emulsion of EPA and DHA in infarcted dogs has acute antiarrythmic properties ⁵' ⁶, the possibility that oxidized metabolites of PUFA are responsible for these beneficial effects is open ¹. Consistently with this hypothesis, an omega-3 PUFA rich diet was recently associated with a higher production of oxygenated metabolites of omega-3 PUFA in the blood circulation ⁸. However the antiarrythmic properties of F₄- neuroprostanes, oxygenated metabolites of DHA, has not yet been investigated in the prior art.

SUMMARY OF THE INVENTION:

DETAILED DESCRIPTION OF THE INVENTION:

The term "cardiac arrhythmia" has its general meaning in the art. Arrhythmias can be classified by rate (physiological, tachycardia, bradycardia), or mechanism (automaticity, reentry, fibrillation). It is also appropriate to classify arrhythmia by site of origin. Accordingly, cardiac arrhythmias include, but are not limited to, ventricular tachycardias including ventricular fibrillation, supraventricular tachycardias, including atrial fibrillation or paroxystic atrial fibrillation. Cardiac arrhythmias are often first detected by the occurrence of pounding, syncope faint, sudden weakness. The simplest specific diagnostic test for assessment of heart rhythm is the electrocardiogram (abbreviated ECG or EKG). A Holier monitor is an EKG recorded over a 24-hour period, to detect arrhythmias that can happen briefly and unpredictably throughout the day.

The F4-NeuroPs could thus constitute a new therapeutic class of anti-arrhythmic drugs that are prescribed to all patients who experienced a myocardial infarction but also patients with other arrhythmic episodes after admission to the hospital.

The study by Calo et al. (2005) (Calo L, Bianconi L, Colivicchi F, Lamberti F, Loricchio ML, de Ruvo E, Meo A, Pandozi C, Staibano M, Santini M. (2005) N-3 Fatty acids for the prevention of atrial fibrillation after coronary artery bypass surgery: a randomized, controlled trial. J Am Coll Cardiol. 2005 May 17;45(10): 1723-8.) demonstrates that PUFA administration 5 days during hospitalization in patients undergoing coronary artery bypass surgery reduced the incidence of postoperative AF (54.4%) and was associated with a shorter hospital stay. Accordingly, the use of F4-NeuroPs of the present invention can thus be envisaged to increase the anti-arrhythmic effect.

Perioperative intravenous infusion of PUFA reduces the incidence of atrial fibrillation after coronary artery bypass surgery and leads to a shorter stay in the hospital. Prolonging the data of Heidt et al. (2009) (Heidt MC, Vician M, Stracke SK, Stadlbauer T, Grebe MT, Boening A, Vogt PR, Erdogan A. (2009). Beneficial effects of intravenously administered N- 3 fatty acids for the prevention of atrial fibrillation after coronary artery bypass surgery: a prospective randomized study. Thorac Cardiovasc Surg. Aug;57(5):276-80.) , perioperative intravenous infusion of F4-NeuroPs of the present invention should be recommended for patients undergoing coronary artery bypass surgery.

The term "F4-neuroprostane" has its general meaning in the art and refers to the class of lipid oxidation metabolites derived from docosahexanoic acid^9"10. In particular, F₄-NeuroPs include but are not limited to 4-F₄-NeuroPs, 7-F₄-NeuroPs, 1 l-F₄-NeuroPs, 10-F₄-NeuroPs, 14-F₄-NeuroPs, 13-F₄-NeuroPs, 17-F₄-NeuroPs and 20-F₄-NeuroPs (Figure 1). F₄-NeuroPs may be synthesised through any method well known in the art. Typically, the compounds may be synthesized by the method described in ref. ^{1 L} In a particular embodiment, the epimers of configuration (S) of the allylic hydroxyl or F₄-NeuroPs with a (S) absolute configuration of the allylic hydroxyl are used.

Typically, the F₄-NeuroPs are administered in a therapeutically effective amount. By a "therapeutically effective amount" is meant a sufficient amount of the F4-NeuroP to prevent or treat cardiac arrhythmias at a reasonable benefit/risk ratio applicable to any medical treatment. It is understood that the total daily usage of the compounds and compositions of the present invention will be decided by the attending physician within the scope of sound medical judgment. The specific therapeutically effective dose level for any particular patient will depend upon a variety of factors including the disorder being treated and the severity of the disorder; activity of the specific compound employed; the specific composition employed, the age, body weight, general health, gender and diet of the patient; the time of administration, route of administration, and rate of excretion of the specific compound employed; the duration of the treatment; drugs used in combination or coincidental with the specific compound employed; and like factors well known in the medical arts. For example, it is well within the skill of the art to start doses of the compound at levels lower than those required to achieve the desired therapeutic effect and to gradually increase the dosage until the desired effect is achieved.

The F4-NeuroP can be used therapeutically in combination with a pharmaceutically acceptable carrier to form pharmaceutical compositions. By "pharmaceutically acceptable" is meant a material that is not biologically or otherwise undesirable, i.e., the material can be administered to a subject, along with the composition, without causing any undesirable biological effects or interacting in a deleterious manner with any of the other components of the pharmaceutical composition in which it is contained. The carrier would naturally be selected to minimize any degradation of the active ingredient and to minimize any adverse side effects in the subject, as would be well known to one of skill in the art. Typically a pharmaceutically acceptable carrier or excipient includes a non-toxic solid, semi- so lid or liquid filler, diluent, encapsulating material or formulation auxiliary of any type.

In the pharmaceutical compositions of the present invention for oral, sublingual, subcutaneous, intramuscular, intravenous, transdermal, local or rectal administration, the active principle, alone or in combination with another active principle, can be administered in a unit administration form, as a mixture with conventional pharmaceutical supports, to animals and human beings. Suitable unit administration forms comprise oral-route forms such as tablets, gel capsules, powders, granules and oral suspensions or solutions, sublingual and buccal administration forms, aerosols, implants, subcutaneous, transdermal, topical, intraperitoneal, intramuscular, intravenous, subdermal, transdermal, intrathecal and intranasal administration forms and rectal administration forms.

Preferably, the pharmaceutical compositions contain vehicles which are pharmaceutically acceptable for a formulation capable of being injected. These may be in particular isotonic, sterile, saline solutions (monosodium or disodium phosphate, sodium, potassium, calcium or magnesium chloride and the like or mixtures of such salts), or dry, especially freeze-dried compositions which upon addition, depending on the case, of sterilized water or physiological saline, permit the constitution of injectable solutions.

The pharmaceutical forms suitable for injectable use include sterile aqueous solutions or dispersions; formulations including sesame oil, peanut oil or aqueous propylene glycol; and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. In all cases, the form must be sterile and must be fluid to the extent that easy syringability exists. It must be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms, such as bacteria and fungi.

Solutions comprising compounds of the invention as free base or pharmacologically acceptable salts can be prepared in water suitably mixed with a surfactant, such as hydroxypropylcellulose. Dispersions can also be prepared in glycerol, liquid polyethylene glycols, and mixtures thereof and in oils. Under ordinary conditions of storage and use, these preparations contain a preservative to prevent the growth of microorganisms.

According to the invention, the F4-NeuroPs can be formulated into a composition in a neutral or salt form. Pharmaceutically acceptable salts include the acid addition salts (formed with the free amino groups of the protein) and which are formed with inorganic acids such as, for example, hydrochloric or phosphoric acids, or such organic acids as acetic, oxalic, tartaric, mandelic, and the like. Salts formed with the free carboxyl groups can also be derived from inorganic bases such as, for example, sodium, potassium, ammonium, calcium, or ferric hydroxides, and such organic bases as isopropylamine, trimethylamine, histidine, procaine and the like.

The carrier can also be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), suitable mixtures thereof, and vegetables oils. The proper fluidity can be maintained, for example, by the use of a coating, such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants. The prevention of the action of microorganisms can be brought about by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like. In many cases, it will be preferable to include isotonic agents, for example, sugars or sodium chloride. Prolonged absorption of the injectable compositions can be brought about by the use in the compositions of agents delaying absorption, for example, aluminium monostearate and gelatin.

Sterile injectable solutions are prepared by incorporating the active polypeptides in the required amount in the appropriate solvent with several of the other ingredients enumerated above, as required, followed by filtered sterilization. Generally, dispersions are prepared by incorporating the various sterilized active ingredients into a sterile vehicle which contains the basic dispersion medium and the required other ingredients from those enumerated above. In the case of sterile powders for the preparation of sterile injectable solutions, the preferred methods of preparation are vacuum-drying and freeze-drying techniques which yield a powder of the active ingredient plus any additional desired ingredient from a previously sterile- filtered solution thereof.

Upon formulation, solutions will be administered in a manner compatible with the dosage formulation and in such amount as is therapeutically effective. The formulations are easily administered in a variety of dosage forms, such as the type of injectable solutions described above, but drug release capsules and the like can also be employed.

For parenteral administration in an aqueous solution, for example, the solution should be suitably buffered if necessary and the liquid diluent first rendered isotonic with sufficient saline or glucose. These particular aqueous solutions are especially suitable for intravenous, intramuscular, subcutaneous and intraperitoneal administration. In this connection, sterile aqueous media which can be employed will be known to those of skill in the art in light of the present disclosure. For example, one dosage could be dissolved in 1 ml of isotonic NaCl solution and either added to 1000 ml of hypodermoclysis fluid or injected at the proposed site of infusion. Some variation in dosage will necessarily occur depending on the condition of the subject being treated. The person responsible for administration will, in any event, determine the appropriate dose for the individual subject.

In addition to the compounds of the invention formulated for parenteral administration, such as intravenous or intramuscular injection, other pharmaceutically acceptable forms include, e.g. tablets or other solids for oral administration; liposomal formulations; time release capsules; and any other form currently used.

The F4-NeuroP of the invention may be formulated within a therapeutic mixture to comprise about 0.0001 to 1.0 milligrams, or about 0.001 to 0.1 milligrams, or about 0.1 to 1.0 or even about 10 milligrams per dose or so. Multiple doses can also be administered. In a particular embodiment, the F4-NeuroP of the invention may be used in combination with another anti-arrhythmic compounds. Typically, the other anti-arrhythmic compounds include but are not limited to Quinidine, Procainamide, Disopyramide, Lidocaine, Phenytoin, Mexiletine, Flecaidine, Propafenone, Moricizine, Propranolol, Esmolol, Timolol, Metoprolol, Amiodarone, Sotalol, Ibutilide, Dofetilide, Verapamil, Diltiazem, Adenosine and Digoxin.

Currently, omega-3 supplementation in people who have suffered a myocardial infarction is used. However, the GISSI study showed that the reduction of mortality after myocardial infarction was 40%. Without being bound by any theory, the inventors believe that the remaining 60% do not have sufficient incorporation of omega-3 FA in the cell membrane and therefore there is no oxygen derivatives, such F₄-NeuroPs in sufficient quantity that can be generated to prevent the fatal arrhythmias. Accordingly a further object of the invention relates to a method for preventing cardiac arrhythmias in a subject in need thereof comprising the steps consisting of i) administering the subject with an omega-3 supplementation ii) determining in a blood sample obtained from the subject the level of F₄-NeuroPs, iii) comparing said level with a predetermined reference level and iv) administering the subject with a F₄-NeuroP according to the invention when the level determined at step i) is lower than the predetermined reference value.

Methods for determining the level of F₄-NeuroPs in a blood sample are well known in the art and typically involved mass spectrometric methods such as GC-MS or LC-MS¹². Briefly, F₄-NeuroPs may be quantified by gas chromatography-mass spectrometry (GC-MS) using electron capture negative ionization and selected ion monitoring.

In one embodiment, the predetermined reference value may be an index value. A predetermined reference value can be relative to a number or value derived from population studies, including without limitation, such subjects having similar body mass index, total cholesterol levels, LDL/HDL levels, systolic or diastolic blood pressure, subjects of the same or similar age range, subjects in the same or similar ethnic group, subjects having family histories of atherosclerosis, atherothrombosis, or CAD, PAD, or CVD. In one embodiment of the present invention, the reference value is derived from the level of F₄-NeuroPs in a control sample derived from one or more subjects who reduction of mortality after myocardial infarction was achieved. In another embodiment, such subjects are monitored and/or periodically retested for a diagnostically relevant period of time ("longitudinal studies") following such test to verify continued absence from fatal arrhythmias. Such period of time may be one year, two years, two to five years, five years, five to ten years, ten years, or ten or more years from the initial testing date for determination of the reference value. Furthermore, retrospective measurement of levels of F₄-NeuroPs in properly banked historical subject samples may be used in establishing the predetermined reference value, thus shortening the study time required, presuming the subjects have been appropriately followed during the intervening period. The invention will be further illustrated by the following figures and examples. However, these examples and figures should not be interpreted in any way as limiting the scope of the present invention.

FIGURES:

Figure 1 depicts the biosynthesis of F₄-neuroprostanes Figure 2: Typical recording with the Ionoptix ® system. The top plot shows the sarcomere length (in micrometer) and the lower the fluorescence ratio F405/F480 is an index of the intracellular calcium concentration. In the absence of isoprenaline (Iso) contractions recorded are very small and the judgment of the stimulation, no arrhythmic event is observed. After application of 10 nM isoprenaline contractions and calcium transients increased in amplitude. To stop the stimulation, an extrasystole (ESV) was observed after 20 seconds.

Figure 3: Plan in vivo experiments where a compound will be retained.

Figure 4: Amount of arrhythmic cells (percentage). Percentage of mice arrhythmic cells in the presence of 10, 100 and 1000 nM 4(RS)-4-F_4t-NeuroP. The IC50 of 4(RS)-4-F_4t- NeuroP seems to be around 100 nM.

Figure 5: Amount of arrhythmic cells (percentage). Percentage of mice arrhythmic cells in the presence of 10, 100 and 1000 nM 10(S)-10-F_4t-NeuroP. The IC50 of 10(S)-10-F_4t- NeuroP appears to be between 10 and 100 nM.

Figure 6: Amount of arrhythmic cells (percentage). Percentage of mice arrhythmic cells in the presence of 10, 100 and 1000 nM 10(R)-10-F_4t-NeuroP. The IC50 of 10(R)-10- F_4t-NeuroP cannot be determined showing the epimer specificity of NeuroP.

Figure 7 shows the general strategy for the synthesis of F₄-NeuroPs

Figure 8 shows the strategy for the synthesis of the 4- and 10- series of F₄-NeuroPs. Figure 9 shows that the effects of DHA, 4(RS)F4t-neuroprostane and carvedilol on arrhythmias triggered by a norepinephrine challenge in post-myo cardial mice (PMI). Mean results from 11 mice in all groups except carvedilol (n=6).***:p<0.001 (Anova one-way followed by Newman- Keuls test).

EXAMPLE: Introduction:

EPA and DHA, chemically unstable due to the presence of several 1,4 pentadienyl groups can be peroxidized. Numerous pathologies, such as the myocardial infarction, are related to the occurrence of a redox imbalance in favor of the production of reactive species of oxygen. Thus, some beneficial effects attributable to DHA and EPA may be due to their oxygenated metabolites. F4-NeuroPs derived from the radical peroxidation of DHA during oxidative stress that accompanies infarction. The effects of F₄-NeuroPs on ionic currents have never been studied, and it is plausible candidates because they are chemically stable.

Material & methods:

Synthesis of the compounds:

Our new strategy used a bicyclo[3.3.0]octene scaffold (1) and focused on F₄-NeuroPs possessing syn-anti-syn stereochemistry¹³. Bicyclo[3.3.0]octene intermediate 1 is easily obtained from 1,3-cyclooctadiene, in 5 steps only, with 18% yield, and in its two enantiomeric forms thanks to an enzymatic resolution. Bicyclo[3.3.0]octene 1 is transformed into 1,5-diol 2 in only few steps. The syn-anti-syn stereochemistry introduced, the later steps of the synthesis involved the side chains introduction and required the desymmetrisation of the two hydroxyl groups. Our strategy bears of flexibility when diol 2 is selectively and enzymatically protected into mono acetate 3.¹⁴ (Figure 7).

Thanks to this synthetically advanced intermediate (3), the synthesis of the 4- and 10- series of F₄-NeuroPs was achieved. Lateral chains were introduced using Wittig, Horner- Wadsworth-Emmons or cross metathesis methodologies. Depending on the nature of the coupling reagent (phosphonium salt, β-ketophosphonate...),

We were able to synthesize the 4- and 10- series of F₄-NeuroPsⁿ by means of this new, flexible and convergent strategy (Figure 8). Study of calcium transients by the photometric system ionOptix ®

a) Principle

Photometric system IonOptix ® can provide real-time, simultaneous acquisition of fluorescence photometry with sarcomere length measurements. For the study of isolated myocytes, the system includes a pacemaker that offers full control of the stimulation pulse duration, frequency and voltage. Thus, the photometric system IonOptix ® puts in correlation the calcium transients and the shortening of the electrically stimulated myocytes (1 Hz, 20 V, Figure 1). Calcium is studied by the use of a ratiometric fluorescent calcium probe, the indol- AM (excitation wavelength 360 ± 10 nm, emission wavelength at 405 ± 10 nm and 485 ± 10 nm).

b) Protocol

To measure the intracellular calcium, the first step is to bring into contact ventricular myocytes load with an amount of indol-AM. To make it liposoluble, Indo-1 is esterified on its carboxylic functions. Once present in the cell, esterases will allow the release of Indo-1 carboxylic functions enabling it to bind to the intracellular calcium. The incorporation time is predetermined to 30 minutes. The incubation time of the probe is critical because too low load generate a weak signal while excessive exposure will excessively increase the buffering capacity of the cytoplasm for calcium.

Once loaded and the cells arranged on the measuring system by means of a tank containing Tyrode 450μί, the probe is excited by a xenon lamp at a wavelength of 360nm. Through an optical wavelength filter, the apparatus collects the fluorescence emitted from the Indo-1 at two different wavelengths: 405 ± 10 nm and 485 ± lOnm. The first one represents the fluorescence of the probe bound to calcium and the second one represents the fluorescence of the free probe. The ratio of these two wavelengths reflects the concentration of intracellular calcium. It is thus possible to observe calcium transients in ventricular myocytes systole but also to observe the diastolic calcium when the stimulation is stopped.

To measure the contraction, we use the striation of cardiac cells due to the presence of sarcomeres. This striation can be scanned and acquired by a computer. The recorded signal is similar to a sine function. This function is then treated mathematically by a Fourier transformation, which allows obtaining the period of the sinusoidal or sarcomere length.

The experimental protocol will address in parallel the changes in intracellular calcium transients and peaks of cell contraction by analysing sarcomere shortening, during 30 seconds of electrical stimulation at 1 Hz, followed by 30 seconds interspersed pause (Figure 1). With this protocol, the goal is to identify arrhythmic events, i.e irregularities in the rate of contractions then to study the effects of omega-3 derivatives on these events. To reproduce the sympathetic tone that characterizes the murine model, the cells are subjected to 10 nM isoproterenol. This molecule in stimulating β-adrenergic system, will then lead to an increase in the amplitude of contractions and calcium transients but also will promote the development of premature ventricular contractions (PVCs) as shown in Figure 2.

Thus, to evaluate the potential anti-arrhythmic of the oxygenated metabolites of DHA, we proceed in two steps:

1 - screening on isolated cells to identify the most effective compounds. Our preliminary results under these conditions suggest anti-arrhythmic properties for several derivatives (Figure 3-4-5) but not all (Figure 6).

2 - After selecting molecules with the highest potential anti-arrhythmic, in vivo experiments in mice will be conducted (in accordance with the ethical European) (Figure 3). The recording of the electrocardiogram (ECG) telemetry allow the identification and counting of ventricular arrhythmias in mice (PMI having undergone ligation of the left main coronary artery). They are similar to those that are sensitive to PUFA in humans. Male mice of 8 weeks (25-30g) will be monitored by Ho Iter established, for the acquisition and subsequent analysis of the ECG in awake mice. Osmotic micropumps are used to chronically deliver the molecules. The animals will be randomly assigned to 4 groups: SHAM (control), Tyrode (PMI control), DHA (PMI receiving DHA) and Oxygenated Metabolites (PMI OM). SHAM group shall be a control to show the effects on the coronary ligation arrhythmias. The DHA group shall be a control to confirm that the intravenous injection of an emulsion of DHA prevents arrhythmia models PMI as observed in the rat and dog.

PMI MO group will be compared to other groups and oxygenated derivatives of them. To estimate the effective concentrations in vivo experiments, circulating concentrations of oxygenated metabolites will be determined.

Results:

The results of the arrhythmic events with 4(RS)-4-F_4t-NeuroP are shown in Figure 4. Other experiments suggest that the S-enantiomer of 10(S)-10-F_4t-NeuroP is more effective than 4(RS)-4-F_4t-NeuroP (Figure 5).

REFERENCES: Throughout this application, various references describe the state of the art to which this invention pertains. The disclosures of these references are hereby incorporated by reference into the present disclosure.

1. Dietary supplementation with n-3 polyunsaturated fatty acids and vitamin E after myocardial infarction: results of the GISSI-Prevenzione trial. GISSI-Prevenzione Investigators Lancet 1999, 354, 447.

2. Cardiovascular effects of marine omega-3 fatty acids. Saravanan P., Davidson N., Schmidt E. & Calder P. Lancet 2010, 376, 540.

3. Dietary long-chain omega-3 fatty acids of marine origin: a comparison of their protective effects on coronary heart disease and breast cancers. Jude, S. Roger, E. Martel, et al, Prog. Biophys. Mol. Biol. 2006, 90, 299.

4. Peroxidation of docosahexaenoic acid is responsible for its effects on ITO and ISS in rat ventricular myocytes. Jude, S. Bedut, S. Roger, et al. Br. J. Pharmacol. 2003, 139, 816.

5. Prevention of sudden cardiac death by dietary pure omega-3 polyunsaturated fatty acids in dogs. Billman G., Kang J. & Leaf A. Circulation, 1999, 99:2452.

6. Acute in vivo administration of a fish oil-containing emulsion improves post- ischemic cardiac function in n-3-depleted rats. Peltier S., Malaisse W., Portois L. et al. Int J Mol Med, 2006, 18, 741.

7. Cardioprotection by omega-3 fatty acids: involvement of PKCs? Le Guennec J-Y.,

Jude S., Besson P. et al. Prostaglandins Leukot Essent Fatty Acids 2010, 82, 173.

8. Dietary polyunsaturated fatty acids and adaptation to chronic hypoxia alter acyl composition of serum and heart lipids. Balkova P., Jezkova J., Hlavackova. et al. Br J Nutr 2009, 102, 1297.

9. F4-isoprostanes: a novel class of prostanoids formed during peroxidation of docosahexaenoic acid (DHA). Nourooz-Zadeh J., Liu E., Anggard E., Halliwell B,. Biochem. Biophys. Res. Com., 1998, 242, 338.

10. Regio chemistry of neuroprostanes generated from the peroxidation of docosahexaenoic acid in vitro and in vivo. Yin FL, Musiek E., Gao L., Porter N. & Morrow J.. J. Biol. Chem. 2005, 280: 2600

11. The handy use of Brown's catalyst for a skipped diyne deuteration: application to the synthesis of a d4-labelled-F4t-neuroprostane. Oger C, Bultel-Ponce V., Guy A., Balas L., Rossi J-C, Durand T., Galano J-M. Chem. Eur. J. 2010, 16, 13976. 12. Quantification of urinary F2-Isoprostanes with 4(RS)-F4t-Neuroprostane as an internal standard using gas-chromatography mass spectrometry : application to polytraumatized patients. Mas E., Michel F., Guy A., Bultel, V., Falquet, Y., Chardon, P., Rossi, J-C, Cristol, J-P., Durand, T. J. Chromato graph. B 2008, 872, 133.

13. Stereocontrolled access to isoprostanes via a bicyclo[3.3.0]octene framework. Oger C, Brinkmann Y., Bouazzaoui S., Durand T., Galano J-M. Org Lett 2008, 10, 5087.

14. Lipase-catalyzed regioselective monoacetylation of unsymmetrical 1,5-primary diols.Oger C, Marton Z., Brinkmann Y., Bultel-Ponce V., Durand T., Graber M., Galano J- M. J Org Chem 2010, 75, 1892.

Claims

CLAIMS:

A method for the treatment and prevention of cardiac arrhythmias in a subject in need thereof comprising administering the subject with a therapeutically effective amount of F₄-neuroprostane (F₄-NeuroP).

The method of claim 1 wherein the F₄-neuroprostane is selected from the group consisting of 4-F₄-NeuroP, 7-F₄-NeuroP, 1 l-F₄-NeuroP, 10-F₄-NeuroP, 14-F₄-NeuroP, 13-F₄-NeuroP, 17-F₄-NeuroP and 20-F₄-NeuroP.

The method of claim 1 or 2 wherein said F₄-neuroprostane is an epimer of (S) configuration of the allylic hydroxyl or is a F₄-neuroprostane with a (S) absolute configuration of the allylic hydroxyl.

The method according to any one of claims 1-3 wherein the F₄-neuroprostane wherein the subject has experienced a myocardial infarction.

The method according to any one of claims 1-4 wherein the cardiac arrhythmias are paroxystic atrial fibrillation or ventricular fibrillations.

The method according to any one of claims 1-3 for reducing the incidence of atrial fibrillation after coronary artery bypass surgery.

A method for preventing cardiac arrhythmias in a subject in need thereof comprising the steps consisting of i) administering the subject with an omega-3 supplementation ii) determining in a blood sample obtained from the subject the level of F₄-NeuroPs, iii) comparing said level with a predetermined reference level and iv) administering the subject with a F₄-NeuroP according to the invention when the level determined at step i) is lower than the predetermined reference value