FR2782270A1 - Use of adenosine and adenylic nucleotides to improve muscular effort in sports and in immobilized and bedridden - Google Patents

Abstract

The use of adenosine, adenylic nucleotides and their salts, in compositions for oral administration to improve muscular work, is new.

Description

ADENOSINE AND ORAL ADENYL NUCLEOTIDES FOR

  IMPROVING MUSCLE WORK

  The present invention relates to the use of adenosine and adenyl nucleotides for the preparation of a composition suitable for oral administration for improving muscle work. Adenylic nucleotides are fundamental constituents of living matter, in particular because they are part of the constitution of nucleic acids (DNA and RNA) responsible for the conservation of biological information. It has also been known for a long time that adenylic nucleotides and, in particular, adenosine 5'-triphosphate (ATP) play an essential role in energy metabolism, since ATP represents the main form of storage of energy required for all of the body's biochemical reactions,

  transmembrane transport of molecules and muscle contraction.

  Since the work of the BURNSTOCK team, it is also well known that adenosine and extracellular adenyl nucleotides are involved in cellular communication as neurotransmitters associated with the sympathetic and parasympathetic systems and as transducers of biological signals. This team was, in fact, the first to demonstrate the existence in the digestive tract of neurons using ATP as a mediator of information transmission (BURNSTOCK, Pharmacol. Rev, 1972, 24, 509-581 BURNSTOCK, Theor. Biol., 1976, 62, 491-503). The discovery of these neurons led to the identification of receptors called purinoreceptors, which are located on the plasma membrane of cells and are made up of proteins composed of 320 to 400 amino acids, some of which are coupled to G proteins (FREDHOLM,

  PharmacologicalReviews, 1994, 46, 143-156).

  There are currently two families of purinoreceptors: P1 purinoreceptors, comprising at least 4 types of receptors (Al, A2a, A2b, A3) and having a preferential sensitivity to adenosine, and P2 purinoreceptors comprising at least 18 types of distributed receptors in families P2X (1-7) and P2y (1-11) and having a preferential sensitivity to ATP. P2y receptors are coupled to G proteins so

  that P2X receptors are ion channels (FREDHOLM, Tips, 1997, 18, 79-82).

  Like most receptors for molecules involved in cell communication, purinoreceptors appear to have a fairly wide distribution. Thus, the purinoreceptors A2a and A2b have a fairly ubiquitous distribution while the A1 receptors are mainly located on adipocytes, cardiomyocytes and and in the brain. P2, x purinoreceptors are located on smooth muscles and the heart and P2y ADP purinoreceptors exclusively on blood platelets. The other purinoreceptors seem to be present on many cell types (brain,

  heart, lung, gastrointestinal tract, smooth muscles, ...).

  The discovery of these purinoreceptors opened the way for a whole

new pharmacology.

  Thus, ATP is used in therapy intravenously in the treatment of paroxysmal tachycardias by reciprocal rhythm of the atrioventricular junction, also known as BOUVERET disease. ATP is also used, but orally, in the treatment of essential back pain. Furthermore, studies, which are reported in Patents US-A-4,880,918 and US-A-5,049,372 in the name of RAPAPORT, have demonstrated an inhibiting effect of ATP on tumor growth and a beneficial action of this. compound on cancer-related cachexia, in favor of possible use in oncology. Finally, ATP is currently the subject of numerous studies aimed at evaluating its interest in the treatment of neurological disorders such as PARKINSON's disease or ALZHEIMER, in the treatment of pain,

  cystic fibrosis, AIDS, and anesthesia.

  However, to date, the fact that the administration of ATP to an organism can have any favorable action on the metabolism of the muscular tissues of this organism appears to have never been described or suggested in the literature. However, in the context of their work, the Inventors have discovered that repeated administration of ATP, orally and at doses infinitely lower than the physiological tissue concentrations of ATP - which we know to be of the order 1.5 g / kg of body weight -, results in both an increase in oxygen supply at the tissue level in general, and at the muscle level in particular, and by a lesser increase in muscle oxygen consumption and in glucose, a lower production of lactic acid and a lowering of thermogenesis during muscular work, thus conferring an interest in the preparation of the organism for muscular effort and in the prevention of the harmful side effects.

  such an effort by improving muscle energy efficiency.

  The subject of the invention is therefore the use of at least one compound chosen from adenosine, adenyl nucleotides and their physiologically acceptable salts for the preparation of a composition suitable for oral administration.

  for improving muscle work.

  Within the meaning of the present invention, the expression "nucleotides

  adenyls "includes adenosine 5'-monophosphate (AMP), adenosine 5'-

  diphosphate (ADP) and adenosine 5'-triphosphate (ATP). Furthermore, the term "physiologically acceptable salts" of adenosine and adenyl nucleotides means all the salts which can be obtained from these compounds and which are compatible with use in vivo. In the case of adenosine, salts which are particularly suitable for use in vivo are the addition salts of this compound with organic acids of the lactic acid, acetic acid, malic acid or citric acid type, or acids inorganics such as hydrochloric acid and phosphoric acid, while in the case of AMP, ADP and ATP, use will preferably be made of addition salts with inorganic bases (soda, potash, ...) or organic bases (ethanolamine, diethanolamine, triethanolamine, phenylethylbenzylamine, procaine ....), these salts possibly being complexed with one or more molecules of water so as to form mono- or

  polyhydrates according to the number of phosphate groups carried by these nucleotides.

  According to the invention, the compound present in the

  composition is preferably adenosine 5'-triphosphate (ATP).

  Advantageously, the compound is present in the composition in an amount suitable for the administration of a dose of said compound of between 1 and 50 mg per day and per kg of body weight, this administration being able to be carried out in

  one or more daily doses.

  Preferably, the amount of compound present in the composition is chosen so as to allow the administration of an amount of said compound of between 1 and 10 mg per day and per kg of body weight. According to a first preferred embodiment of the use in accordance with the invention, the composition is a pharmaceutical composition comprising the compound as active ingredient, and at least one excipient

pharmaceutically acceptable.

  According to another preferred embodiment of the use in accordance with the invention, the composition is a dietetic composition suitable for use as a nutritional supplement and comprising the compound as

  active substance, and at least one physiologically acceptable vehicle.

  In accordance with the invention, such a composition, whether of a pharmaceutical or dietetic nature, can also comprise amino acids and, in particular, essential amino acids (valine, leucine, isoleucine, lysine, methionine, threonine, phenylalanine, tryptophan), proteins, sugars with slow or rapid assimilation, unsaturated or polyunsaturated fatty acids and / or lipids containing such fatty acids such as phospholipids, vitamins (A, Bl, B2, B6, C, E, PP, ...), mineral salts (calcium, phosphorus, magnesium,

  .), trace elements (zinc, selenium, manganese, molybdenum, ...) or any other compound..DTD: to improve the metabolism of muscle tissue.

  Furthermore, this composition can be in all forms suitable for oral administration, such as tablets, capsules, capsules, sucking or chewing tablets, solutions or suspensions which are drinkable or even under the

  form of powders to dissolve in a liquid.

  The composition prepared in accordance with the invention finds numerous applications. Thus, for example, it is likely to be used by athletes, whether professional, regular or occasional athletes, with a view to improving their performance and their tolerance to muscular effort in the part of a training or sporting event. It is also likely to be used by force workers (loggers, movers, soldiers from operational units, firefighters, ...) wishing to obtain a reduction in muscle fatigue linked to the exercise of their professional activity. Finally, it is likely to be prescribed to immobilized or bedridden patients as an adjunct treatment, in order to increase their resistance in the context of functional rehabilitation (physiotherapy, physiotherapy, etc.). This composition is, moreover, of interest for the prevention of the harmful consequences secondary to a muscular effort such as cramps and muscular pains, muscular exhaustion or hyperthermia

of effort.

  The invention will be better understood using the complement of

  description which follows, given as an example of demonstration of beneficial action

  of oral and repeated administration of ATP on muscle metabolism, and with reference to the appended drawings in which: - Figure 1 illustrates, in the form of a histogram, the effects of oral administration of ATP, d '' a duration of 14 days and at the daily dose of mg / kg of body weight (series 1), and those of an administration of distilled water carried out under the same conditions (control series), on the partial arterial and venous pressures oxygen at rest; FIG. 2 illustrates, in the form of a histogram, the effects of an oral administration of ATP, lasting 14 days and at the daily dose of mg / kg of body weight (series 1), and those of an administration of distilled water carried out under the same conditions (control series), on the total quantities of oxygen present in the arterial blood and in the venous blood in a situation of rest; - Figure 3 illustrates, in the form of curves, the effects of an oral administration of ATP, for a period of 14 days and at the daily dose of mg / kg of body weight (e), and those of a administration of distilled water carried out under the same conditions (o), on the intramuscular temperature during muscular effort; - Figure 4 illustrates, in the form of curves, the effects of an oral administration of ATP, for a period of 14 days and at the daily dose of mg / kg of body weight (e), and those of a administration of distilled water carried out under the same conditions (o), on the muscular oxygen extraction (or consumption) during muscular effort; Figure 5 illustrates, in the form of curves, the effects of an oral administration of ATP, lasting 14 days and at the daily dose of mg / kg of body weight, and those of a administration of distilled water carried out under the same conditions (o), on muscle consumption of glucose during muscular effort; - Figure 6 illustrates, in the form of curves, the effects of an oral administration of ATP, lasting 14 days and at the daily dose of mg / kg of body weight (e), and those of administration of distilled water carried out under the same conditions (o), on the muscular production of lactic acid; and - Figure 7 illustrates, in the form of curves, the effects of an oral administration of ATP, lasting 14 days and at the daily dose of mg / kg of body weight (e), and those of '' a distilled water administration carried out

  under the same conditions (o), on the metabolic acidosis of the venous blood.

  It should be understood, however, that this complement of

  description is given solely by way of illustration of the object of the invention and does not

  in no way constitutes a limitation.

  EFFECTS OF ORAL AND REPEATED ATP ADMINISTRATION ON

MUSCLE METABOLISM

  The beneficial action of oral and repeated administration of ATP on muscle metabolism has been established by a series of experiments consisting in treating New Zealand rabbits orally with ATP for 14 days and assessing the effect of such treatment: - on the one hand, on oxygenation parameters in a resting situation, namely the arterial and venous partial oxygen pressures (PaO2 and PvO2) and the total quantities of oxygen (bound oxygen hemoglobin + dissolved oxygen) present in arterial and venous blood; and on the other hand, on metabolic and oxygenation parameters during a muscular effort, that is to say on: a muscular thermogenesis (or heat production by the muscles), by measuring the internal temperature of the muscles put in condition of muscular work, the extraction (or consumption) muscular in oxygen, by the measurement of the total quantities of oxygen (oxygen linked to hemoglobin + dissolved oxygen) present in the arterial blood and in the venous blood, for muscle consumption of glucose, by measuring plasma glucose levels in arterial blood and venous blood, muscle production of lactic acid, by measuring plasma lactate levels in arterial blood and venous blood, and to metabolic acidosis in venous blood, by measuring venous pH; and to compare the results obtained with those observed, under the same conditions, in "control" rabbits, that is to say rabbits having been treated, orally and during

14 days, with distilled water.

  1 - Experimental protocol All the experiments were carried out on male rabbits, weighing approximately 3.5 kg, housed and tested in accordance with the GUIDE FOR THE CARE AND USE OF LABORATORY ANIMALS published in 1996 by the NATIONAL RESEARCH COUNCIL. These rabbits were systematically put in

  water diet the day before the experiments.

  To assess the oxygenation parameters, the rabbits were anesthetized with sodium pentobarbital, then tracheotomized to allow the measurement and recording of the ventilatory parameters (frequency, resistance) by the pneumotachograph PR 800 and the PMS software (Pulmonary Monitoring System ) 800 sold by NUMED. A catheter has been placed in the tracheal tube to provide oxygen-enriched ventilation and maintain pressure

  arterial physiological oxygen.

  A catheter connected to a pressure cell and provided with a sampling site was introduced into the left carotid until the aortic arch for on the one hand, allowing the measurement and recording of aortic pressures and, on the other On the other hand, perform the arterial samples required to measure the arterial partial oxygen pressure (PaO2) and the total amount of oxygen present in the arterial blood. Similarly, a catheter with a sampling site was placed in a collateral of the femoral vein draining the gastrocnemius, i0 plantaris, soleus and vastus lateralis muscles to carry out the venous sampling necessary for measuring partial pressure. venous (PvO2) and the total amount of oxygen present in the venous blood. An electrocardiogram has also been put in place

  in leads D I, D II and D III to control the heart rate.

  Once the stability of the preparations has been verified by measuring the various physiological parameters (ventilatory parameters, aortic pressures, heart rate), the arterial samples intended for the measurement of PaO2 and PvO2 as well as the total quantities of oxygen present in the arterial blood. and in

  venous blood was taken every 10 minutes for 2 hours.

  The assessment of the metabolic and oxygenation parameters under muscular effort was carried out on rabbits prepared in the same manner as previously, but in which the sciatic nerve of the left paw was isolated and a bipolar electrode was placed on this nerve. The muscles innervated by the sciatic nerve, namely the gastrocnemius, the plantaris, the soleus and the vastus lateralis, were put into working condition for 45 minutes by stimulation of the sciatic nerve at a frequency of 1.3 hertz, at an intensity 40 milliamps, the duration of each

  stimulation being 0.1 millisecond.

  The internal temperature of the muscles was recorded during the whole

  duration of experiments using thermometric probes.

  The arterial and venous samples required to measure the total quantities of oxygen present in the arterial blood and in the venous blood, the glucose and lactate contents of the arterial blood and the venous blood as well as the pH of the latter were respectively performed by means of catheters placed in the aortic arch and in the collateral of the femoral vein draining the stimulated muscles. These arterial and venous samples were taken every 15 minutes during the 45 minutes of stimulation of the sciatic nerve. The total amount of oxygen, the plasma glucose and lactate contents, as well as the pH in the case of venous blood, have

  were measured on the same sample.

  In all experiments, the total quantities of oxygen available in arterial blood and / or venous blood were established according to the method

  de ROSSING and CAIN (J AppliedPhysiol., 1965, 21, 1, 195-201).

  2 - Results a) PaO2, PvO2 and total quantities of oxygen present in arterial blood and in venous blood in a resting situation Figure 1 represents, in the form of a histogram, the mean + standard deviation of partial pressures arterial (non-hatched bar) and venous (hatched bar) oxygen, expressed in mm of mercury, as measured on 2 series of 12 rabbits each, having received daily by the oral route, i.e. 20 mg of ATP per kg of body weight ( series 1), i.e. distilled water (control series) for 14 days, while Figure 2 shows, also in the form of a histogram, the mean + standard deviation of the total quantities of oxygen present in the blood arterial (non-hatched bar) and in venous blood (hatched bar), expressed in ml per 100 ml of

  blood, as measured on these same 2 series of rabbits.

  As can be seen in Figures 1 and 2, oral and repeated administration of ATP results, in a resting situation, in a significant increase in PaO2 (P <0.001 compared to the control series) and in the total amount of oxygen available in arterial blood (21.64 + 0.493 ml of O2 per 100 ml of blood for series 1 versus 21.10 0.339 ml of O2 per 100 ml of blood for the control series), testifying to a greater oxygen supply at the tissue level in general, and at the muscle level in particular. At the same time, PvO2 and the total amount of oxygen in the venous blood (13.90 + 0.381 ml of O2 per 100 ml of blood for series 1 versus 14.03 + 0.565 ml of O2 per 100 ml of blood for the control series) appear to be comparable in rabbits treated with ATP and rabbits in the control series, signifying a slightly higher muscular oxygen consumption, in

  resting situation in treated rabbits.

  b) Intramuscular temperature in situation of muscular effort Figure 3 represents, in the form of curves, the mean + standard deviation of the differences, expressed in C, as a function of the time between the internal temperatures of the muscles put in working condition as described in point 1 above and the initial internal temperatures of these muscles, as measured on 2 series of 12 rabbits each, having received daily orally 20 mg

  ATP per kg of body weight (a), i.e. distilled water (o), for 14 days.

  As shown in this Figure 3, the oral and repeated administration of ATP significantly results in a less increase in the internal temperature of the muscles put into working condition, reflecting a lower production of heat by these muscles during of this work and advocating for a

  better muscular endurance during exercise.

  c) Muscle oxygen extraction under muscular effort Figure 4 represents, in the form of curves, the mean + standard deviation of the muscle oxygen extraction coefficients (E) as a function of time as established from measurements of the total quantities of oxygen present in arterial blood and in venous blood carried out during muscular work as described in point 1 above, on 2 series of 12 rabbits each, having received daily orally 20 mg of ATP per kg of body weight, i.e.

  distilled water (o), for 14 days.

  The oxygen extraction coefficients (E) were established using the following formula: Total amount of arterial O2 - Total amount of venous O2

  (E) = ---------------------------------------------- ------------------------------ x 1 0O.

  Total amount of arterial O2 As shown in Figure 4 and in agreement with the experimental data presented in Figures I and 2, muscle oxygen consumption is, at the time when muscle work begins (point 0 of the axis abscissa), significantly higher in rabbits treated with ATP than in untreated rabbits. Then, while the muscles of untreated rabbits rapidly contract, in response to effort, an oxygen debt resulting in a marked increase in the oxygen extraction coefficient (E), the muscles of rabbits treated with ATP do not contract this oxygen debt and the oxygen extraction coefficient (E)

  increases little in these rabbits treated in response to stress.

  d) Muscle consumption of glucose in a situation of muscular effort Figure 5 represents, in the form of curves, the mean + standard deviation of the differences, expressed in mmol / liter of blood, as a function of the time between the plasma glucose contents in arterial blood and in venous blood as measured, during muscular work as described in point 1 above, on 2 series of 12 rabbits each, having received daily orally 20

  mg ATP per kg bodyweight (e), i.e. distilled water (o), for 14 days.

  As shown in this Figure 5, the oral and repeated administration of ATP results, at the end of a muscular effort, by a less increase in

  glucose consumption by muscles.

  e) Muscle production of lactic acid in situations of muscular effort

  Figure 6 shows, in the form of curves, the mean + deviation-

  type of the differences, expressed in mmol / liter of blood, as a function of the time between the plasma lactate contents in the venous blood and in the arterial blood as measured, during muscular work as described in point 1 above , on 2 series of 12 rabbits each, having received daily orally or 20 mg

  ATP per kg of body weight (e), i.e. distilled water (o), for 14 days.

  As illustrated in this Figure 6, repeated oral administration of ATP results in less production of lactic acid by the muscles put in

Condition of work.

  f) Metabolic acidosis in venous blood under muscular effort Figure 7 represents, in the form of curves, the mean + standard deviation of the venous pH values as measured, during muscular work as described in point 1 above, on 2 series of 12 rabbits each, having received daily by the oral route, i.e. 20 mg of ATP per kg of body weight (a),

  or distilled water (o), for 14 days.

  As can be seen in this Figure 7, repeated oral administration of ATP results in a lesser increase in metabolic acidosis in the venous blood during muscular effort, which is linked to the lower production of acid.

  lactic acid from the muscles during this effort.

  As is apparent from the above, the invention is in no way limited to those of its modes of implementation and application which have just been described; on the contrary, it embraces all the variants which may come to the mind of the technician in the matter, without departing from the framework or the scope of this

Invention.

Claims (10)

  1. Use of at least one compound chosen from adenosine, adenylic nucleotides and their physiologically acceptable salts for the preparation of a composition suitable for oral administration for improving muscle work. 2. Use according to claim 1, wherein the compound is adenosine 5'-triphosphate.   3. Use according to claim 1 or claim 2, wherein the compound is present in the composition in an amount suitable for the administration of a dose of said compound of between 1 and 50 mg per day per kg body weight.   4. Use according to claim 3, wherein the compound is present in the composition in an amount suitable for the administration of a dose of said   compound between 1 and 10 mg per day per kg of body weight.   5. Use according to any one of claims 1 to 4, in   which the composition is a pharmaceutical composition which comprises the compound as active ingredient, and at least one pharmaceutically excipient acceptable.   6. Use according to any one of claims 1 to 4, in   which the composition is a dietary composition suitable for use as a nutritional supplement and comprising the compound as an active substance, and   at least one physiologically acceptable vehicle.   7. Use according to any one of claims 1 to 6, in   wherein the composition further comprises one or more compounds chosen from amino acids, proteins, sugars with slow or rapid assimilation, unsaturated or polyunsaturated fatty acids and / or lipids containing such fatty acids, vitamins,   mineral salts and trace elements.   8. Use according to any one of claims 1 to 7, in   which the composition is intended to improve performance and / or tolerance to muscular effort.   9. Use according to any one of claims 1 to 7, in   which the composition is intended to serve as an adjunct treatment in patients immobilized or bedridden.   10. Use according to any one of claims 1 to 7, in   which the composition is intended for the prevention of harmful consequences   secondary to muscular effort.