JPS6354690B2 - - Google Patents

Description

[Detailed description of the invention]

[Industrial Application Field] The present invention relates to the combination of a ubiquinone coenzyme such as coenzyme Q ₁₀ and creatine phosphate or N
-(phosphonamidino)sarcosine (N-
The present invention relates to a pharmaceutical composition containing as an active ingredient a combination of phosphorylated organic compounds known as phosphonamidino and sarcosine. [Summary of the Invention] The combined components exhibit a strong synergistic effect and have a surprising and unexpected effect in improving muscle metabolism and in particular in protecting the heart from damage caused by anoxia. The results were derived. The damage caused by the anoxia can occur, for example, during myocardial infarction or post-infarction.
infarction), during cardiovascular surgery, or when muscular energy demands are increased. The surprising pharmacological and therapeutic effects exerted by the pharmaceutical compositions of the present invention are due to coenzyme
This is due to the synergistic effect that occurs between Q ₁₀ and creatine phosphate. In fact, Coenzyme Q ₁₀ is known to play a very important role in the respiration process of cells and tissues (Gale PH et al., Archives of Biochemistry and Biophysics). Arch.Biochem.Biophys.), vol. 93, p. 211, 1961
Year). Coenzyme Q ₁₀ intervenes in mitochondrial electron transport and oxygen utilization by the cytochrome respiratory chain (Morton RA)
Nature, volume 182, page 1764,
(1959). Coenzyme Q ₁₀ is present in almost all tissues, especially muscle, heart, brain and liver, and coenzyme Q ₁₀ deficiency is associated with diseases such as myocardial infarction, myocardial failure, muscular dystrophy and arterial hypertension in the above organs. A decrease in the concentration of coenzyme Q ₁₀ has been demonstrated in some pathological conditions of these organs, suggesting that a relationship exists between them. In fact, exogenous administration of coenzyme _Q10 was found to reverse the reduced concentration of this coenzyme in tissues and at the same time exhibit important therapeutic effects (Folkers K. et al. International Journal of Vitamin Research (Int.J.Vit.Res.), Volume 40, Page 380,
1970). Many pathological conditions such as myocardial failure, post-infarction conditions, myopathic dystrophies and hypertension have been successfully treated by the administration of Coenzyme Q ₁₀ (Yamasawa's Biomedical and Clinical Aspects of Coen Team Q ₁₀ (Biomedical and
Clinical Aspects of Coenzyme Q ₁₀ ), Elsevier, North Holland
Holland), vol. 333, 1980). Further biochemical and pharmacological studies showed that administration of coenzyme _Q10 , besides restoring decreased mitochondrial respiration, also reduced cellular ATP.
It has been demonstrated that cell membranes can be maintained intact by increasing the concentration. A similarly important role is played by creatine phosphate. The critical concentration of mitochondrial ATP, as occurs through cytochrome chains and coenzyme _Q10 , is
Essential for the production of creatine phosphate. The creatine phosphate is synthesized within the mitochondria by mitochondrial ATP and cytoplasmic creatine. The molecule is capable of transporting high-energy phosphate bonds to myofibrils and cell membranes. In fact, at this level creatine phosphate is used by creatine phosphokinase to produce ATP from ADP and to maintain muscle contraction and ion transport. Several experiments have shown that the tissue content of creatine phosphate is a factor that controls the strength of heart contractions. Administration of creatine phosphate prolongs the myocardial contractile function of rat hearts during reversible ischemic arrest (Hearse DJ et al., Journal of Pharmacy and
Pharmacology (J.Pharm.Pharmac.), vol. 26,
427, 1974), reducing the incidence of premature ventricular contractions caused by coronary ligation (Marshall RJ and Parrat JR, Naunyn; Schmiedebergs', Archives of Pharmacol., vol. 281, 437.
Page, 1974). Furthermore, a decrease in intracellular phosphate at high energy levels was found to lead to damage to the sarcolemma and constitute the beginning of irreversible ischemic damage. As shown in studies conducted in ischemic rat hearts or in experimental myocardial infarctions in dogs, administration of creatine phosphate can prevent depletion of higher phosphates and inhibit processes that lead to breakdown of muscle cell membranes. can. The study showed that administration of creatine phosphate not only significantly improved cardiac function and high-energy phosphate levels, but also increased the integrity of myocardiocyte sarcolemma and concomitantly Degradation of membrane phospholipids led to a decrease in lysophospholipids (Saco et al.
“Journal” by Sako VA et al.
of Molecular and Cellular Cardiology
Cardiology, 16, Supplement 2 September 1984'', Sharov VG, Journal of Cardiovascular Pharmacol., 1984
Fagliemi O., Journal of Cardiovascular Pharmacology, Vol. 4, p. 53, 1982). Results obtained from studies of the myocardial protective effect of creatine phosphate have also been demonstrated in the clinic, in acute myocardial infarction, demonstrating a marked reduction in the occurrence of ventricular arrhythmias, and in cardiovascular surgery. Treated patients showed more rapid recovery of hemodynamic and electrocardial activity, with preservation of normal concentrations of high-energy phosphates and obvious loss of ischemic damage to the sarcolemma, as confirmed by right ventricular biopsies. A protective effect was observed (Robinson LA, Haas DA).
J.O.'s "Journal of Therapy Cardiovascular Surgery"
Therapy Cardiovascular Surgery), Volume 87, 190
Page, 1984). It has now been found that the pharmaceutical compositions of the present invention provide extremely surprising pharmacological and therapeutic effects due to the synergistic interaction between the ubiquinone component and creatine phosphate. Such a synergistic effect would not be expected based on what is known so far or from a simple addition of the effects of single components. [Effects and Examples] The results of pharmacological and toxicological tests conducted on the pharmaceutical composition of the present invention will be described below. Coenzyme Q ₁₀ and creatine phosphate are known to have very low toxicity, and no change in toxicity was observed when they were combined. In fact, the combination of those two ingredients is 1:
Subcutaneous or intravenous injections into mice and rats of solutions containing a ratio of 10 to 1:100 (ie, much higher amounts than would be expected for therapeutic purposes) did not cause any toxic effects. Under such conditions, the LD ₅₀ was found to be greater than 1 g/Kg and 500 mg/Kg for subcutaneous and intravenous administration, respectively. Chronic toxicity studies in rats conducted by oral administration of the above combination in a 1:10 ratio for 30 consecutive days also revealed no pathological changes in body weight and various blood chemistry parameters. It was not made clear. (1) Protective effect against ventricular arrhythmia induced by coronary ligation in rats Selyck et al. (Angiology) for surgical occlusion of the left coronary artery
11, p. 398, 1960) and Clark C. et al. (J. Pharmacol.
Methods, Vol. 3, p. 357, 1980) was applied to male rats weighing 350-450 g. Rats that showed the appearance of arrhythmia or a decrease in arterial pressure below 75 mm Hg during surgery and before ligation were excluded. Coenzyme Q ₁₀ and Creatine Phosphate Solution, Coronary Artery Ligation
A slow injection was given into the left ventricular lumen 15 minutes prior. Arrhythmia began to appear 4-7 minutes after ligation, and ectopic ventricular contractions were counted for 30 minutes after ligation. Coenzyme Q ₁₀ and creatine phosphate can reduce the number of ectopic contractions caused by coronal ligation, but this effect is significantly and surprisingly enhanced by their combination, but As clearly shown in Table 1, there is a significant synergistic effect.

【table】

[Table] (2) Effect on ATP content in papillary muscles of rabbits after hypoxia A group of New Zealand rabbits with an average weight of 2 kg were treated with a solution containing 10 mg/Kg of coenzyme Q ₁₀ or 100 mg/Kg of creatine phosphate for 3 days. This was followed by intravenous treatment. Other groups of animals were treated with a combination of the two drugs at the same dose or half the dose. All animals, including controls, were sacrificed 2 hours after the last injection, the hearts were removed, and equal sections of papillary muscle 1 mm in diameter and 4-5 mm thick were isolated. The tissue was analyzed in a suitable thermostatic bath perfused with a 100% saturated _O2 solution. Hypoxia was created by introducing 100% _N2 into the bath instead of _O2 . ATP content in papillary muscle was determined by Strehler BL et al. ( Methods in Engineering
in Enzymology, New York, Academic Press, 871 pages,
(1957). The results obtained (Table 2) are for the Koen team.
It has been shown that only in the case of the combination of Q ₁₀ and creatine phosphate the ATP content is maintained at normal values even after hypoxia, thus 2
It clearly demonstrates the synergistic effect between the two compounds.

[Table] (3) Effect on enzyme activity in gastrocnemius muscle of rats subjected to daily muscle exercise (training) Considering the fact that daily training in rats increases mitochondrial enzyme activity in both the heart and gastrocnemius muscle. The increase depends on the duration of training and muscular performances (Benzi G. et al., Journal of Applied Physiology, vol. 38, p. 565).
(1975), the combination of the invention was tested for any shortening effect on muscle movement and therefore on the enzymatic order of functional movement. Although injections of creatine phosphate or coenzyme _Q10 were found to increase the mitochondrial enzymatic pool in rat gastrocnemius muscle, a combination of the two substances was shown to have an even higher effect. In this case, the increase in mitochondrial enzyme activity after 7 days of treatment was higher than that obtained after 30 days of training. The animals (Wistarats) were trained on a Rotarod track (Rotarod equipment Basile, Comerio, Italy);
Exercise activity was performed daily for 120 minutes at 20 m/min. Enzyme activity was determined after 30 days of training.
The gastrocnemius muscle was evaluated by isolating and homogenizing (Oscai L.
B.) Journal of Biological Medicine (J.Biol.Med.), vol. 246, p. 6968,
1971). The results obtained (Table 3) show that by administering the pharmaceutical composition of the invention, the increase in mitochondrial enzyme activity is even greater than that caused by training. , such results are not obtained when the two components are used separately.

[Table] Yes.
The effective dose for humans of the pharmaceutical composition of the present invention is coenzyme Q ₁₀ 0.10-8.5 mg/Kg body weight + creatine phosphate 0.10-4.5 mg/Kg body weight. The pharmaceutical composition of the present invention includes capsules, sugar-coated pills,
May be in the form of gastroresistant tablets, vials or ampoules for injection;
The active ingredient may also be inglobated in liposomes or lipidic microspheres. The pharmaceutical compositions of the present invention may also be prepared in a reservoir plug for immediate dissolution in a suitable Coenzyme Q ₁₀ solution.
The active ingredient may be formulated in ampoules or vials for immediate dissolution containing creatine phosphate sodium salt in lyophilized form and possible excipients in a plug).

Claims (1)

[Scope of Claims] 1. A heart and muscle metabolism protective agent comprising a ubiquinone coenzyme and creatine phosphate or a salt thereof. 2. The heart and muscle metabolism protecting agent according to claim 1, wherein the ubiquinone coenzyme is coenzyme _Q10 . 3. Claim 1, wherein the weight ratio of coenzyme Q ₁₀ :creatine phosphate is 1:1000 to 1:2.
The cardiac and muscle metabolism protective agent according to item 1 or 2. 4. Solid, semi-solid or liquid for oral, parenteral or rectal administration for the treatment of myocardial anoxia and muscular energetic and metabolic abnormalities damage in the treatment of humans and/or animals. Claims 1 and 2 in the form
3. The heart and muscle metabolism protective agent according to item 1 or 3. 5. The heart and muscle metabolism protecting agent according to claim 4, which is in the form of a capsule, a sugar-coated pill, a stomach-resistant tablet, a vial or an ampoule for injection. 6. The heart and muscle metabolism protecting agent according to claim 1, 2 or 3, wherein the active ingredient is spherically contained in liposomes or lipid globules. 7. Valid in ampoules or vials for immediate dissolution containing creatine sodium phosphate sodium salt in lyophilized form and possible excipients in a reservoir plug for immediate dissolution in a suitable Coenzyme Q ₁₀ solution. The heart and muscle metabolism protecting agent according to claim 1, 2 or 3, wherein the components are formulated.