Anti-Stress, Anti-Impairment and Anti-Aging Drug And

Home , Adafenoxate, Azaperone, Etifoxine, Isopropyl alcohol, Meclofenoxate, Pramiracetam

~™ llll III II III I II III III 1 1 II I II (19) J European Patent Office

Office europeen des brevets (11) EP 0 804 239 B1

(12) EUROPEAN PATENT SPECIFICATION

(45) Date of publicationation and mention (51) |nt. CI.6: A61 K 45/06 of the grant of the patent: 15.09.1999 Bulletin 1999/37 (86) International application number: PCT/RO94/00003 (21) Application number: 94919023.5 (87) International publication number: (22) Date of filing: 02.06.1994 WO 95/33486 (14.12.1995 Gazette 1995/53)

(54) ANTI-STRESS, ANTI-IMPAIRMENT AND ANTI-AGING DRUG AND PROCESS FOR MANUFACTURING THEREOF ARZNEIMITTEL GEGEN STRESS, BEEINTRACHTIGUNG UND ALTWERDEN UND VERFAHREN ZU IHRER HERSTELLUNG MEDICAMENT ANTI-STRESS, ANTI-DEFICIENCE ET ANTI-VIEILLISSEMENT ET SON PROCEDE DE FABRICATION

(84) Designated Contracting States: • Riga, Sorin AT BE CH DE DK ES FR GB GR IE IT LI LU MC NL 71248 Bucharest (RO) PTSE Designated Extension States: (74) Representative: SI Petra, Elke, Dipl.-lng. et al Petra, Zieger & Kollegen (43) Date of publication of application: Patentanwalte 05.11.1997 Bulletin 1997/45 Herzog-Ludwig-Strasse 18 85570 Markt Schwaben (DE) (73) Proprietors: • Riga, Dan (56) References cited: 71248 Bucharest (RO) • ANNALS OF THE NEW YORK ACADEMY OF • Riga, Sorin SCIENCE, vol.717, 30 June 1994 pages 315 - 331 71248 Bucharest (RO) Predescu V et al 'Antagonic-stress: A new treatment in gerontopsychiatry and for a healthy (72) Inventors: productive life' • Riga, Dan 71248 Bucharest (RO)

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O Note: Within nine months from the publication of the mention of the grant of the European patent, give CO any person may notice to the European Patent Office of opposition to the European patent granted. Notice of opposition shall be filed in o a written reasoned statement. It shall not be deemed to have been filed until the opposition fee has been paid. (Art. Q_ 99(1) European Patent Convention). LU Printed by Xerox (UK) Business Services 2.16.7/3.6 EP 0 804 239 B1

Description

[0001] The invention relates to an anti-stress, anti-impairment and anti-aging drug for prophylactic, therapeutic and restorative use; process for the manufacture of the drug and to the use of the drug according to claims 1 - 3 for the man- 5 ufacture of a medicament for prophylactic medicine and for the manufacture of a medicament for therapeutics and restorative medicine for stress-dependent pathology and in accelerated aging. [0002] Especially it relates to an orally-administered drug having anti-stress, anti -impairment and anti-aging etio-pathogenic actions, and manifold therapeutic efficiency, as a result of its synergistic biological, neurometabolic and cell- trophic composition. The invention also relates to a process for its manufacturing. This drug supports and enhances the 10 adaptation capability and the neuropsychic and biological resistance, simultaneously achieves the anti-stress protection of the brain, liver and heart, and corrects the main ultrastructural and metabolic imbalances brought about by acute and chronic stress, prolonged biological wear and tear, oxidative stress, ischemia-hypoxia, chronic alcoholism and premature aging, thus being meant for stressdependent pathology. The non- conventional pharmaceutical process for its manufacturing further provides the controlled guidance in the release and absorption of the active substances, and the 15 prolonged cerebral vasodilative action, which together promote maximum bioavailability and therapeutic efficacy.

Background Art

[0003] Acute and chronic stress, a negative and permanent characteristic of present-day life, comprises the unfolding 20 of three processes: first - the aggression of the body by stressors, in a continuous growth, diversification and perpetu- ation, second - the body response, which may be adaptively, maladaptively or pathologically stress-dependent; and third - the intense, accelerated and chronically accumulated wear and tear of the brain and of the organism, premature aging. [0004] The adaptive response of the body (the general adaptation syndrome) consists of three successive stages: 25 immediate adaptation, long-term adaptation and the stage of exhaustion - neuropsychic and biological impairment (SELYE, H., The evolution of the stress concept, American Scientist 61 , 692-699, 1973). Chronic wear and tear means the neuropsychic and biological progressive incapacitation: it results in the decrease of the adaptation capability and in the diminution of body vitality and resistance, and it is the consequence of accumulation in the course of time of stress- ful life events and stress-induced lesions. When the adaptation capabilities of the organism are exceeded, due to the 30 intensification, frequency and chronicization of stress and impairment, diseases of adaptation appear, meaning the stress-dependent pathology: neuropsychiatric and psychosomatic illnesses (WILDER, J.F., PLUTCHIK, R., Stress and psychiatry (Cap. 25.1 1), pp. 1 198-1203, In: KAPLAN, H.I., SADOCK, B.J. (Eds.), Comprehensive Textbook of Psychia- try/IV, vol. 1, Williams and Wilkins, Baltimore, 1985). [0005] The human brain, through its triple functionality - neurobiological, psychic and social - and due to the loss, after 35 birth, of the neuron regeneration capacity through cell division, represents the global "receiver" of stresses (ischemic, hypoxic, oxidative etc.) and the "storage" of chronic, progressive, neuropsychic and biological impairment. Thus, in the central nervous system, specific ultrastructural and biochimical imbalances and lesions are accumulated, reaching all levels of metabolism (energy, anabolism, catabolism), then they extend with age and determine the incapacitation of the brain and body functions. The cerebral blood flow diminution as a consequence of stress and aging (progressive 40 chronic hypoxia) and the neuronal hypoanabolism (the disturbance and decrease of nucleic acid and protein synthesis, the reduction and impairment of Nissl bodies - crowds of rough endoplasmic reticulum and free ribosomes - and of Golgi apparatus) induce the diminution of plasticity and anabolic regeneration, both functional (enzymes and neurotransmitters) and ultrastructural (neurosomes and extensions), (TERRY, R.D., GERSHON, S. (Eds.), Aging, vol. 3 (Neu- robiology of Aging), Raven Press, New York, 1976). Oxidative stress, neuronal hypercatabolism, lipid peroxidation, 45 especially of membranes, and the premature chronic impairment of subcellular organelles, mainly mitochondria, in all cases finally result in a progressive accumulation of lipofuscin pigments (wear and tear pigments, age pigments, tertiary lysosomes, insoluble subcellular wastes coming from peroxidation, polymerization and cross-linkages by free radicals) in neurons and glial cells (RIGA, D., RIGA, S., POPESCU, A., CONSTANTINESCU E., PERIETEANU, M., Subcellular genesis of the nerve lipofuscin pigments, 4th European Anatomical Congress, Basle, Switzerland, 1977; Acta Anatom- 50 ica, 99, 307-308, 1977; ZS.-NAGY, I. (Ed.), Lipofuscin - 1987: State of the Art, Academiai Kiado, Budapest, 1988). [0006] The necessity to develop a specific drug having an etio-pathogenic action against stress, impairment and premature aging was determined by the profound negative consequences of stress:

a) at the individual level - professional failure, disease, premature aging and death, and 55 b) at the level of the whole society - important economic and social losses, direct and indirect (COOPER, C, ARBOSE, J., Executive stress goes global, International Management, 39, 42-48, 1984).

[0007] From the prior art it can be seen that efforts made for the production of drugs efficient in the control and treat-

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ment of stress and impairment have so far failed to produce a specific drug with an etio-pathogenic action; only symp- tomatological purposes or energizing effects have been fulfilled. [0008] Thus, for improving the symptomatology induced by stress, dysadaptation, maladaptation responses to stress and the stress-related disorders (anxiety, depression, asthenia, sleeplessness, neurotic, neurovegetative and psycho- 5 somatic disorders) the use of psychotropic medication is known (POLDINGER, W., SCHMIDILIN, P.E., WIDER, R, Index Psychopharmacorum, H. Huber, Bern, 1983). Depending on the prevailing symptomatology, the following are known as having been used:

a) anxiolytics (minor tranquilizers): diazepam, meprobamate, methylpentynol, etifoxine; 10 b) neuroleptics(major tranquilizers):chlorpromazine, promethazine, azaperone; c) beta-adrenergic blockers : bunitrolol; d) antidepressants: tricyclic, tetracyclic compounds, alone or associated with neuroleptics; e) psychostimulants: caffeine, amphetamine or derivatives; f) sedatives and hypnotics: combining in the formula phenobarbital and codeine (Romanian Patents nos. 60376 15 and 64161).

[0009] These psychotropic drugs have the disadvantage of representing only a predominant symptomatic medication, without an etio-pathogenic action against stress; they do not reduce chronic impairment caused by stress, they do not act against stress through anabolic regeneration, they modify the normal (anti-stress) reactions of body adaptation and 20 bring about numerous adverse reactions. Furthermore, the anxiolytic and psychostimulant drugs (of the amphetamine and caffeine type) often call for increasing posology, due to phenomena of acquired tolerance and determine, as important adverse reactions, the dependence on psychoactive substances (LADER, M., Benzodiazepines - the opium of the masses?, pp. 609-615, In: SMITH, A.D., LLINAS, R., KOSTYUK, RG. (Eds.), Commentaries in the Neurosciences, Per- gamon Press, Oxford, 1980; W.H.O. Europe, Prevention of Mental, Psychosocial and Neurological Disorders in the 25 European Region, 38th Session, Copenhagen, 12-17 September, 1988). [001 0] One also knows many anti-stress drug compositions used for energizing, activatory stimulating, trophic, tonic, fortifying - neuropsychic and/or biological - purposes. They are employed in treating disorders caused by acute and chronic stress, by stress-dependent pathology and especially against their most frequent consequences: nervous, psychic and biological exhaustion, accelerated chronic impairment, premature senescence. 30 a) Some of these compositions contain polyvitamins: hydrosoluble (American Medical Association, AMA Drug Evaluations, Publishing Sciences Group, Acton, Mass., 1973), hydrosoluble together with liposoluble (U.S. Patent no. 3493659), or polyvitamins with bioelements (French Patent no. 7404 M); b) Other mixtures associate amino acids with or without vitamins (acetylaspartic acid, arginine glutamate, citrulline 35 with folic acid), or are based on aspartates (Romanian Patents nos. 55069 and 77472; French Patent no. 2521 429), on glutamates (Romanian Patent no. 76141), or cysteine (Romanian Patent no. 74505), arginine (French Patent no. 2494113), or complex combinations usually built up, alongside with the above mentioned amino acids, from gly- cine, lysine, tyrosine, ornithine, histidine (French Patent no. 5937 M; Romanian Patent no. 76044); c) Other associations comprise stimulating substances and combinations thereof: amphetamine, amphetamine 40 with caffeine, or caffeine with vitamins (Romania Patents nos. 62137 and 66014).

[001 1 ] These products, even when they are complex drug associations, have the drawback of not achieving an etio- pathogenic anti-stress therapy by simultaneous coupling of some multiple (vasodilative, normolipidemic, energo-active, anti-toxic, of catabolic regulation - lipofuscinolysis and anabolic regeneration) actions; they are not preclinical^ tested 45 in antagonization of the experimental stress induced in animals, they cannot prevent or decelerate nervous wear and tear by antagonization of the oxidative stress and do not support the natural adaptation mechanisms - anti-stress mechanisms. Furthermore, especially drugs that associate methylxanthines and/or amphetamines are disadvantageous, because they form an incomplete, limited medication, of short-term effects, which in case of chronic administration, or in large doses, determine other new imbalances, contributing to the enhancement of those pre-existent, to the diminu- 50 tion of neuropsychic and biological resistance to stress, intensifying cellular catabolism and maladaptive reactions against stress; moreover, their use induces many adverse reactions (SYED, I.B., The effects of caffeine, Journal of the American Pharmaceutical Association, NS 16, 568-572, 1976; IVERSEN, LL, IVERSEN, S.D., SNYDER, S.H. (Eds.), Handbook of Psychopharmacology, vol. 11 (Stimulants), Plenum Press, New York, 1978). [0012] The use of methionine in hepatic pathology, as a hepatoprotective compound is well known (WADE, A., REY- 55 NOLDS, J.E.F. (Eds.), The Extra Pharmacopoeia, The Pharmaceutical Press, London, 1978). We found out by successive investigations that methionine additionally has a specific neurotropic action and interferes etio-pathogenically against stress. In this way we demonstrated the antagonization effect of chronic oxidative stress, determined by the diminution of lipofuscin pigments in the brain (telencephalon and diencephalon) of old rats, using quantitative histo-

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chemical methods (RIGA, S., PAMBUCCIAN, G., OERIU, S., Changes in lipofuscin pigments of rat central nervous system under -SH groups' releasing substances' influence, 9th International Congress of Gerontology, vol. 3 (Section Session, Abstracts), abstract no. 1 103, p. 383, Kiev, U.S.S.R., 1972). Subsequently, we emphasized its complex action anti-stress, anti-impairment and anti-aging (catabolic regulation - anti-lipofuscinogenesis, lipofuscinolysis, and anabolic 5 regeneration - the increase of RNA synthesis) at the nervous system level of old rats, using a complex methodology: selective isolations of living nerve cells from brain, morphometrical and biochemical methods (RIGA, S., Studies on Nucleic Acids in the Central Nervous System in Senescence Processes, Ph. D. Thesis, Institute of Medicine and Phar- macy, Bucharest, 1976, in Romanian). [001 3] It is also known that meclofenoxate, thanks to its actions of energetic and metabolic regulation on nerve cells, 10 has a wide-spread area of clinical use in psychiatry, neurology as well as in the pathology determined by the hypoxic stress aggression of the brain - geriatrics, neurosurgery, anesthesiology, intensive care (COIRAULT, R., DELIGNE, P., ROUIF, J., Une orientation therapeutique nouvelle. L'A.N.P. 235 (ester dimethyl-amino-ethylique de I'acide para-chlorophenoxy-acetique), Agressologie, 1 ,1 13-138,1960, in French). Afterwards, using light microscopy-histochemistry (qualitative only), the property of meclofenoxate to decrease lipofuscin pigments in the nervous system of old guinea pigs 15 was emphasized (NANDY, K., BOURNE, G.H., Effect of centrophenoxine on the lipofuscin pigments in the neurons of senile guinea-pigs, Nature (Lond.), 210, 313-314, 1966); it was also demonstrated by electron microscopy (HASAN, M., GLEES, P., EL-GHAZZAWI, E., Age-associated changes in the hypothalamus of the guinea pig: effect of dimethylami- noethyl p-chlorophenoxyacetate. An electron microscopic and histochemical study, Experimental Gerontology, 9, 1 53- 159, 1974). Our researches, having priority as preclinical methodology (quantitative - morphometry and qualitative - 20 type of distribution, autofluorescence, histochemistry) and our trivalent experimental pattern (statistical comparison of three groups - control young, control old and treated old) have demonstrated the meclofenoxate action of specific and intense decrease of lipofuscin pigments in the brain of old rats, based on that, the authors of the present invention introduced in the scientific literature in the art the concept of lipofusciolysis (RIGA, S., RIGA, D., Effects of centrophenoxine on the lipofuscin pigments in the nervous system of old rats, Brain Research, 72, 265-275, 1974), sustaining it also by 25 electron microscopy (RIGA, D., RIGA, 10 S., Selektive lipofaszinolytische Effekte von Centrophenoxin am Nervensys- tem alter Ratten, pp. 22-27, In- KUGLER, J. (Ed.), Himstoffwechsel und Himdurchblutung, Schnetztor Verlag, Konstanz, Schweiz, 1977). Later on this concept was used by other authors too. As a result of its lipofuscinolytic action, proof of the antagonization of oxidative stress, meclofenoxate acts etio-pathogenically against brain stress, impairment and aging (RIGA, S., RIGA, D., Dynamics of lipofuscin pigments, directing factor of brain aging, 6e Congres Medical Inter- 30 national de la Federation Internationale des Resistants (F.I.R.), Prague, Tchecoslovaquie, le 30 novembre - 2 decembre 1976, Resumes, p. 70, 1976) and in deceleration of 20 aging rate (ZS.-NAGY, 1., An attempt to answer the questions of theoretical gerontology on the basis of the membrane hypothesis of aging, Advances in the Biosciences, 64, 393 - 413, 1987). [001 4] The two above mentioned substances, administered separately or without an anti-stress potentiation by means 35 of a neurometabolic composition with an etio-pathogenic action, are disadvantageous because they offer only an incomplete protection of the brain against stress, impairment and aging, while their actions of functional and metabolic regulation, as well as of subcellular regeneration are only partial. [001 5] It is therefore the problem of the invention, to provide a restorative drug that overcomes the deficiencies of the state of the art. 40 [001 6] The problem is solved by a drug according to claim 1 , the process for ist production according to claim 4 and to ist use for the manufacture of a medicament in restorative medicine according to claims 7 and 8. [0017] The drug with anti-stress, anti-impairment and anti-aging etio-pathogenic action, according to the invention, carries out a synergistic biological, neurometabolic and cell-trophic composition, elaborated in a specific anti-stress therapeutic conception, by association and synergism of the following active principles: 45 a) anti-oxidative and anti-catabolic stress compounds (anti-lipofuscinogenesis. lipofuscinolytics and for lipofuscin elimination): methionine plus aminoethanol phenoxyacetates and/or aminoethyl phenoxyacetamides); b) anti-anabolic stress components (for functional and ultrastructural anabolic regeneration): hydrooxopyrimidine carboxylates and/or oxopyrrolidine acetamides with potassium, zinc and lithium; so c) vasodilators (anti-hypoxic and anti-ischemic stress) and normolipidemic compounds: nicotinic alcohol and/or acid or derivatives thereof with magnesium and iodine; d) energo-active and e) anti-toxic components: aspartate, fructose, vitamin B-|, vitamin B6, monoacid phosphate and sulfate.

55 [0018] The process for drug manufacturing, according to the present invention, in order to fulfill in oral administration a maximum bioavailability and therapeutic efficiency, stipulates:

a) for controlled delivery in the release and absorption of the active compounds from the drug, the pharmaceutical

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preparation of the composition in two complementary types of capsules or film-coated tablets (gastrosoluble and enterosoluble units), the latter being enteric-coated, and b) for achieving the prolonged anti-ischemic and anti-hypoxic actions, the nicotinic alcohol or acid, or its derivatives from the composition of the enterosoluble capsule or film-coated tablet is retarded, under the form of granule or tab- 5 let.

[001 9] The drug, according to the invention, offers the following advantages:

it represents a new drug and a new pharmacotherapeutical strategy in the fields of health promotion and in the 10 prophylaxis, control and treatment of stress, biological wear and tear, premature aging, stress-related illnesses; it was elaborated and tested according to the biological drug conception, carrying out a synergistic biological, neurometabolic and cell-trophic (cerebral, hepatic, myocardial and general) composition, resulting in a multiple therapeutic efficacy, without toxicity and adverse reactions, without tolerance and dependence; it was created and checked up in a specific therapeutic conception, namely etio-pathogenic and homeostatic, 15 against stress, impairment and aging, thus ensuring higher therapeutic efficiency, based on its active constituents (anti-oxidative and anti-catabolic stress, anti-anabolic stress, vasodilative - anti-hypoxic and anti-ischemic stress - and normolipidemic, energo-active and anti-toxic compounds), in free radical pathology, hypercatabolic and hypo- anabolic states, in hypoxic-ischemic pathology, in dyslipidemias, in energetic exhaustion and toxic-deficient pathology; 20 - its biological composition and its specific anti-stress action determine a large area of prophylactic, therapeutic and recovering uses, for the whole scale of ages (child, young man, adult, old man), for both healthy and sick persons; it is meant for healthy people, especially during activities with a high stressant and performing coefficient, because it antagonizes the anti-homeostatic action of stress and impairment (at neuropsychic and biological levels) and enhances the working power in overstressing conditions through increasing the resistance to psychic (intellectual) 25 and physical (biological) effort; it is meant for patients in stress-dependent, psychosomatic, psychiatric and neurologic pathology, in geriatrics and internal medicine.

[0020] The administration of the drug, may efficiently contribute to the maintainance and improvement of the mental 30 and biological state of health, to a significant reduction of economic and social losses, determined in the whole of society by stress, biological impairment, premature aging and by the stress-dependent pathology. [0021] The pharmaceutical procedure for manufacturing the drug, according to the present invention, offers the following advantages:

35 - in case of oral administration, it allows maximum bioavailability and therapeutic efficiency; it ensures controlled guidance in the release and absorption of active substances from the drug, by formulating the composition in two complementary types of pharmaceutical units (capsules or film-coated tablets): gastrosoluble and enterosoluble; for the enteric coating of the enterosoluble unit, it employs a technologic procedure completely non-toxic and non- 40 polluting, namely the spraying with aqueous dispersion of gastroresistant-enterosoluble polymer; it ensures a prolonged or sustained anti-ischemic and anti-hypoxic action by timed-release of the vasodilative component from the enterosoluble unit; the different colouring of the two complementary units, gastro- and enterosoluble, and the drug package in the form of calendar-blister, in usual conditions under silica gel protection or under vacuum package technology, ensures 45 correctness, easiness, differentiation and evidence (daily, weekly, monthly) of the treatment and the suitable pre- servability of the drug; thanks to its dosage flexibility, it makes possible the differentiation of anti-stress and anti-imparment treatment depending on individual features and neuropsychic reactivity, as well as on obtaining at brain level mainly the anti- oxidative and anti-catabolic stress or anti-anabolic stress action; so - it offers the possibility to administer the drug by chronotherapeutic criteria (chronobiological posology).

Best Mode for Carrying Out

[0022] Below are presented six examples of pharmaceutical production of the drug, according to this invention. For 55 the purpose of controlled delivery in the release and absorption of the active substances, we established a selective formulation of the drug composition in two complementary units, gastro- and enterosoluble. The two complementary units are manufactured either in capsules, or in the form of film-coated tablets, and they are coloured differently for their identification in oral administration. In each of these examples are used the necessary quantities of active compounds

5 EP 0 804 239 B1 and adjuvants in order to prepare 100 capsules or film-coated tablets. [0023] The examples are given on illustrative purpose only and they do not limit in any way the scope of the invention.

Example 1

[0024]

A) The composition for 100 gastrosoluble units is the following:

- DL Methionine 13.50 g - Meclofenoxate hydrochloride (2-(Dimethylamino)ethyl(p-chlorophenoxy)acetate hydrochloride 24.00 g - Zinc sulfate, anhydrous 0.90 g - Nicotinic acid with immediate-release 0.90 g - DL Aspartic acid • Mg salt, anhydrous 8.00 g - D(-) Fructose 0.90 g - Vitamin B-| hydrochloride 0.70 g - Vitamin B6 hydrochloride 1 .00 g

B) The composition for 100 enterosoluble units is as follows:

- Orotic acid, anhydrous (1 ,2,3,6-Tetrahydro-2,6-dioxo-4-pyrimidinecarboxylic acid) 29.00 g - di-Potassium hydrogen phosphate, anhydrous 8.80 g - Lithium carbonate 0.20 g - Nicotinic acid with prolonged-release 7.00 g - Magnesium oxide 1 .80 g - Potassium iodide 0.01 g - D(-) Fructose 0.90 g

[0025] A minimum active anti-stress dose, for one day, consists in the administration of two gastrosoluble units and one enterosoluble unit.

A) The process for manufacturing the gastrosoluble unit. The above mentioned composition for 100 gastrosoluble units may be formulated in two preferable ways, namely: a mixture of granules and powders, or a film-coated tablet and powders.

a) The preparation of the mixture composed of granules of meclofenoxate and powders of the other active substances is performed in conformity with the physico-chemical properties of the active principles and using the method of successive dilutions, thus obtaining six mixtures (l-VI), as follows:

I 24.00 g meclofenoxate hydrochloride [20], over which 15.00 g isopropyl alcohol in thin film are poured, are granulated through the sieve 10 with medium thread and dried at 35°C. The granule thus obtained, after drying, is homogenized through the sieve 20, then are powdered with 0.277 g aerosil [90] and briquet- ted with a 9 mm diameter punch. The resulting briquette is grinded on the sieve 20; II 0.90 g anhydrous zinc sulfate [20], 0.90 g D(-) fructose [20] are separately homogenized by mixing, then they are incorporated into the following mixture: III made up of 0.90 g nicotinic acid [20], 0.70 g vitamin B-| hydrochloride [20] and 1 .00 g vitamin B6 hydro- EP 0 804 239 B1

chloride [20]; IV obtaining the mixture IV (IV = II + III); V 24.277 g mixture I [20], 13.50 g DL methionine [20] and 8.00 g anhydrous DL aspartic acid • Mg salt, [20] are homogenized, forming a mixture; VI in the previous mixture V, the mixture IV is progressively incorporated, and the whole is powdered with 0.549 g talc [90], thus resulting the final mixture VI (the composition for 100 gastrosoluble units).

The simple numbers or numbers in brackets represent the codes of German Standard Specifications of the sieves for each row material of pharmaceutical technologies. b) The preparation of the mixture consisting in the meclofenoxate film-coated tablet and the powders of the other active substances is performed in two stages. In the first step, the film-coated tablet of meclofenoxate is manufactured. After granulating the meclofenoxate, according to the technique described above, except the stage of briquetting, it is compressed on a lenticular punch, 7 mm in diameter, and afterwards the tablet is coated with a gastrosoluble polymer film. The second step consists in the preparation of the other mixtures, according to the technique described under a).

For the pharmaceutical preparation of the active substances one may also use other known adjuvants that can achieve the technological and therapeutic purposes of the drug. The gastrosoluble unit composition, prepared according to procedures a) or b), is encased in hard gelatin (operculated) capsules as follows:

a) the filling of capsules with granules and powders is performed in one stage, using a single dosage station (the one used for granules-powders), and is followed by their closing; b) the filling of capsules with tablet and powders is performed in two stages, using two successive dosage sta- tions (one for tablets and the other for powders) and is followed by their closing. In both procedures, the oper- ations of filling and closing the capsules are carried out automatically by filling and closing machines, in harmony with GMP (Good Manufacturing Practices) guidelines.

The gastrosoluble composition may also be encased in soft gelatin capsules. Furthermore, for the pharmaceutical preparation of the gastrosoluble unit one may employ other procedures of production, like the compression of active substances and adjuvants, followed by the film coating of the tablet obtained with a gastrosoluble polymer in aqueous dispersion, for instance hydroxypropyl methylcellulose dispersion of 3 cP and 5-6 cP viscosity, methacrylic copolymer or a gastrosoluble polymer disolved in an organic solvent, e.g. hydroxypropyl methylcellulose of 15 cP and 50 cP, 3 cP and 5-6 cP viscosity, methacrylic copolymer. In all above mentioned cases, the gastrosoluble unit of drug (gastrosoluble capsule or film-coated tablet) is obtained, in conformity with the pharmaceutical requirements (Romanian Pharmacopoeia (RO. Ph. X), Xth ed., Medical Publishing House, Bucharest, 1993, in Romanian; European Pharmacopoeia (Eur. Ph.), 2nd ed., Maison- neuve, Sainte-Ruffine, vol. 1, 1980 and vol. II, nos. 1-13, 1980-1990; The United States Pharmacopoeia (USP XXII), XXII rev, The United States Pharmacopeial Convention, Rockville, Md., 1990; British Pharmacopoeia (BP 1988), vol. 1 and vol. 2, Her Majesty's Stationery Office, London, 1988). B) The process for manufacturing the enterosoluble unit. The novelty of the process, according to the invention, consists in the production of an enterosoluble unit, being able to ensure controlled guidance in the release and absorption of the active substances, as well as the prolonged anti-ischemic and anti-hypoxic action. The two medical necessities are solved by the process, according to the invention, but the pharmaceutical technology is performed in reverse order. Thus, the new pharmaceutical process provides the timed-release of nicotinic acid as a vasodilative active principle and the enteric coating of capsules, after their filling and closing, resulting in their gastroresistance and enterosolubilization. For retardation, the nicotinic acid is prepared either in the form of a prolonged-release granule, uniformly embedded in other powder mass, or as a prolonged-release tablet, introduced together with the rest of powders in a capsule.

a) In the following, the preparation of the composition for 100 enterosoluble units, with the timed-release granule of nicotinic acid is described. For retardation, in the first step ethyl cellulose as insoluble polymer and diethyl phthalate as plasticizer in a ratio (wt/wt) of 1/0.18-1/0.24 are used, and in the second step Carnauba wax as retarder is introduced in a ratio (wt/wt) of 1/0.47-1/0.53 to the first retarder. In preparing the content of capsules, achieved by the method of successive dilutions, four mixtures (l-IV) are obtained, as follows: EP 0 804 239 B1

I over 7.000 g nicotinic acid [20], 0.430 g aerosil [90], 0.262 g polyvinylpyrrolidone [90] forming a homogeneous mixture, a solution of 4.080 g isopropyl alcohol, 3.000 g methylene chloride and 0.178 g diethyl phthalate in which 0.870 g ethyl cellulose was dissolved on a water bath, under stirring, is added in thin film. The mass thus obtained is granulated through the sieve 10 with medium thread, dried at 35°C, and then the granule is made uniform on the sieve 10 with medium thread. After that, a solution of 2.000 g isopropyl alcohol, 1 .300 g methylene chloride, in which 0.434 g Carnauba wax was dissolved on a water bath, under stirring, is added. The mixture is granulated on the sieve 10 with medium thread, dried at 35°C, then uniformized on the sieve 10 with medium thread; II 0.01 g potassium iodide [45], 0.001 g sodium thiosulfate [45], 0.20 g lithium carbonate [45] in a homogeneous mixture are incorporated in 0.90 g D(-) fructose [45]; III 1 .80 g magnesium oxide [45] are added to the mixture II, then they are homogenized by repeated sifting through sieve 10; IV in a homogeneous mixture of 29.00 g anhydrous orotic acid [20] with 8.80 g anhydrous dipotassium hydrogen phosphate [20], the mixture III is incorporated, then the mixture I is added, and finally is powdered with 0.2628 g aerosil [90]. Thus, the final mixture IV (the composition for 100 enterosoluble units) is obtained.

The simple numbers or numbers in brackets represent the codes of German Standard Specifications of the sieves for each row material of pharmaceutical tehnologies. b) The manufacturing of the composition for 100 enterosoluble units, with the timed-release tablet of nicotinic acid takes place in two stages. The first step consists in obtaining the prolonged-release tablet by compressing the retarded granules of nicotinic acid according to the technique described above, on a lenticular punch, 6 mm in diameter. The second step consists in preparing the other mixtures according to the technique described under a).

The two modalities allow a differentiated timed-release of the nicotinic acid, during a period of about 2-3 hours in the case of procedure a) and in an interval of about 7-8 hours in the case of procedure b). The composition of the enterosoluble unit, prepared by processes a) or b) is introduced into hard gelatin (operculated) capsules, in similar conditions as those described for filling and closing the gastrosoluble capsules. The enterosoluble composition may be encased in soft gelatin capsules too. As it was already mentioned above, another novelty of the process is the enteric coating of hard gelatin capsules. The preferred technology is the enteric coating in aqueous dispersion, because this is economic and at the same time conforms with antipolluant standards. The process of achieving the enteric coating is that of the f luidized bed, which allows the uniform coating of the capsules on their whole surface. One of the preferred compositions of the enteric coating dispersion for 100 capsules is the following:

- Cellulose acetate phthalate 5. 1 60 g - Diethyl phthalate 1 .840 g - 25 % Ammonium hydroxide 1 .500 g - Colorant 0.006 g

The enterosoluble polymer/plasticizer ratio (wt/wt) is 1/0.33-1/0.39, and the enterosoluble polymer/colorant ratio (wt/wt) is 1/0.0010-1/0.0012. Other compositions of the aqueous dispersion for enteric coating, used in the process according to the invention, may contain, instead of cellulose acetate phthalate other gastroresistant-enterosoluble polymers, like: polyvinyl acetate phthalate, hydroxypropyl methylcellulose phthalate, cellulose acetate trimethylate, methacrylic copolymer. Although it was indicated that, for enteric coating, aqueous dispersions of the above mentioned polymers are preferred, they can be used solved in organic solvents too. As plasticizer, instead of diethyl phthalate, one can also employ dibutyl phthalate, propylene glycol, triacetin. The plasticizers used improve flexibility and durability of the coating increase resistance to chipping or cracking and decrease the transmission of water vapours through this film. The economic, non-polluting process and the industrial equipment of good yield, accu- racy and reproducibility used for enteric coating in aqueous dispersion, according to the present invention, fit the qualitative guidelines of GMR The enteric coating of the capsules is preferably performed on an equipment of intermittent film coating, which

8 EP 0 804 239 B1

ensures the proper drying of the resulting film, and avoids any film irregularities, capsule adhesion or capsule diformity. Likewise, the enteric coating can also be made by means of a continous film coating instalation, taking into account that drying times have to increase in direct proportion to the thickness of the film, in order to obtain its optimum drying and uniformity. Due to the sensitivity of gelatin capsule wall to aqueous solutions, the coating pro- 5 cedure has two stages. The first consists in a pre-coating with a thin layer of enterosoluble polymer film, up to the weight of about 15-20 mg/capsule. In this phase a maximum air to a minimum polymer ratio in the aqueous dispersion must be maintained, to obtain the rapid drying of the film. The average temperature at which the coating takes place is 70° C. In the second stage, using spraying, the gradual increase in the film thickness and weight is carried out, by changing the above mentioned ratio, meaning the increase of the polymer aqueous dispersion flow to the 10 detriment of the air flow, concomitantly with the gradual increase in drying times, depending on the thickness of the film obtained. The second step lasts until reaching a weight of about 60-70 mg film/capsule. The optimum weight is established in accordance with the result of the disintegration test, according to Eur. Ph., 1980: gastroresistance > 2 hours, enterosolubilization < 1 hour. Furthermore, for the pharmaceutical preparation of the enterosoluble unit one may employ other production 15 processes, like the compression of the active substances alter the retardation of nicotinic acid, using well-known adjuvants, followed by the film coating of the tablet thus obtained with a gastroresistant-enterosoluble polymer in aqueous dispersion or solved in an organic solvent. In all above mentioned cases, the enterosoluble unit of the drug is obtained (enterosoluble capsule or film- coated tablet), which corresponds to the stipulations of RO. Ph. X, 1993, Eur. Ph., 1980, USP XXII, 1990, and BP, 20 1988.

Example 2

[0026] 25 A) The composition for 100 gastrosoluble units is the following:

30 - DL Methionine 16.50 g - Meclofenoxate hydrochloride (2-(Dimethylamino)ethyl (p-chlorophenoxy)acetate hydrochloride) 28.00 g - Zinc sulfate, anhydrous 1 .20 g - Nicotinic acid with immediate-release 1 .10 35 3g - DL Aspartic acid • Mg salt, anhydrous 1 0.00 g - D(-) Fructose 1.10 g - Vitamin B-| hydrochloride 0.90 g 40 - Vitamin B6 hydrochloride 1 .20 g

In this formulation, meclofenoxate hydrochloride may be replaced by meclofenoxate hybenzate, within the 45 same limits. B) The composition for 100 enterosoluble units is as follows:

50 - Orotic acid, anhydrous (1 ,2,3,6-Tetrahydro-2,6-dioxo-4-pyrimidinecarboxylic acid) 34.00 g -di-Potassium hydrogen phosphate, anhydrous 10.80 g -Lithium carbonate 0.30 g - Nicotinic acid with prolonged-release 9.00 55 g - Magnesium oxide 2.20 g - Potassium iodide 0.01 4g

9 EP 0 804 239 B1

(continued) D(-) Fructose 1.10 g

In this formulation, orotic acid may be used both in its anhydrous form and as monohydrate, within the same limits. A minimum active anti-stress daily dose, consists in the administration of two gastrosoluble units and one enterosoluble unit. The pharmaceutical technology of the two complementary units develops in accordance with the stages described in example 1 .

Example 3

[0027]

A) Examples 1 and 2 not being limitedly, for manufacturing the gastrosoluble unit, out of the class of aminoethanol phenoxyacetates, the 24.00 g meclofenoxate hydrochloride may be replaced by 16.00 g meclosulfonate hydrochloride, as can be seen from this example, which also achieves the therapeutic purposes of the drug. In this case, the composition for 100 gastrosoluble units is the following:

- L(-) Methionine 13.50 g - Meclosulfonate hydrochloride (2-(Dimethylamino)ethyl(o-chloro-p-diinethylamidosulfonylphenoxy) ace- 16.00 g tate hydrochloride) - Zinc sulfate, anhydrous 0.90 g - Nicotinamide (Vitamin PP) with immediate-release 0.90 g - L(+) Aspartic acid • Mg salt, anhydrous 8.00 g - D(-) Fructose 0.90 g - Vitamin B-| mononitrate 0.70 g - Vitamin B6 hydrochloride 1 .00 g

B) Examples 1 and 2 not being limitedly, for manufacturing the enterosoluble unit, the hydrooxopyrimidine carboxylates may be replaced by oxopyrrolidine acetamides, in the following ratio (wt/wt): 1/1 .48-1/1 .60, calculated for one unit of orotic acid. In this way, the 29.00 g anhydrous orotic acid are replaced by 45.50 g piracetam, as can be seen from this example, which also carries out the therapeutic purposes of the drug. In this case, the composition for 100 enterosoluble units is as follows:

- Piracetam (2-Oxo-1 -pyrrolidine acetamide) 45.50 g - di-Potassium hydrogen phosphate, anhydrous 8.80 g - Lithium carbonate 0.20 g - Nicotinic alcohol with prolonged release 6.50 g - Magnesium oxide 1 .80 g - Potassium iodide 0.01 g - D(-) Fructose 0.90 g

In the same way, the 45.50 g piracetam can be replaced by other compounds from the class of oxopyrrolidine EP 0 804 239 B1

acetamides, like:

- 1 8.00 g Oxiracetam (4-Hydroxy-2-oxo-1 -pyrrolidine acetamide); - 1 3.00 g Etiracetam ((+)-alpha-Ethyl-2-oxo-1 -pyrrolidine acetamide); - 7.00 g Dimethylphenyl piracetam (N-[2-(2,6-Dimethylphenyl]-2-oxo-1 -pyrrolidine acetamide); - 20.50 g Pramiracetam (N-[2-(Diisopropylamino)ethyl]-2-oxo-1 -pyrrolidine acetamide); - 23.00 g Aniracetam (1 -p-Anisoyl-2-pyrrolidinone).

A minimum active anti-stress dose, for one day, is represented by the administration of two gastrosoluble units and one enterosoluble unit. The pharmaceutical technology of the two complementary units develops in the same way as that described in example 1 .

Example 4

[0028]

A) Following the procedure in example 3, 20.00 g meclosulfonate hydrochloride can be used instead of the 28.00 g meclofenoxate hydrochloride, as described below. Accordingly, the composition for 100 gastrosoluble units is the following:

- L(-) Methionine 16.50 g - Meclosulfonate hydrochloride (2-(Dimethylamino)ethyl(o-chloro-P-dimethylaminosulfonylphenoxy) ace- 20.00 g tate hydrochloride) - Zinc sulfate, anhydrous 1 .20 g - Nicotinamide (Vitamin PP) with immediate-release 1 .05 g - L(+) Aspartic acid • Mg salt, anhydrous 10.00 g -D(-)Fructose 1.1 Og - Vitamin B-| mononitrate 0.90 g -Vitamin B6 hydrochloride 1 .20 g

B) Following the procedure in example 3, 51 .20 g piracetam can be used instead of the 34.00 g anhydrous orotic acid, as described below.

-Piracetam (2-Oxo-1 -pyrrolidine acetamide) 51 .20 g - di-Potassium hydrogen phosphate, anhydrous 10.80 g - Lithium carbonate 0.30 g - Nicotinic alcohol with prolonged-release 7.90 g - Magnesium oxide 2.20 g - Potassium iodide 0.01 4g - D(-) Fructose 1.10 g

In its turn, the 51 .20 g piracetam can be replaced by other compounds from the class of oxopyrrolidine acetamides, like: EP 0 804 239 B1

- 22.00 g Oxiracetam; - 17.00 g Etiracetam; - 8.00 g Dimethylphenyl piracetam; - 24.50 g Pramiracetam; - 27.00 g Aniracetam.

A minimum active anti-stress daily dose, consists in the administration of two gastrosoluble units and one enterosoluble unit. The pharmaceutical technology of the two complementary units develops according to the steps described in example 1 .

Example 5

[0029]

A) Examples 1 -4 not being limitedly, for producting the gastrosoluble unit, from the class of aminoethanol phenoxyacetates, the 24.00 g-28.00 g meclofenoxate hydrochloride may be also replaced by:

- 19.00 g - 23.00 g Eclofenoxate hydrochloride (2-Diethylamino)ethyl(p-chlorophenoxy)acetate hydrochloride); - 5.00 g - 7.00 g Adafenoxate (2-(1 Adamantylamino) ethyl(p-chlorophenoxy)acetate),

which also achieve the therapeutic purposes of the drug. As an illustration, bellow we present the replacement of the 24.00 g meclofenoxate hydrochloride by 19.00 g eclofenoxate hydrochloride. Thus, the composition for 100 gastrosoluble units is the following:

-L(-) Methionine 13.50 g - Eclofenoxate hydrochloride (2-(Diethylamino)ethyl(p-chlorophenoxy)acetate hydrochloride 19.00 g - Zinc sufate, anhydrous 0.90 g - Nicotinic alcohol with immediate-release 0.80 g - DL Aspartic acid • Mg salt, anhydrous 8.00 g -D(-) Fructose 0.90 g - Vitamin B-| hydrochloride 0.70 g - Vitamin B6 hydrochloride 1 .00 g

Within the framework of aminoethanol phenoxyacetate class, the whole or partial replacement of meclofenoxate by its therapeutic homologues is made within the range of the following ratios (wt/wt):

- Meclofenoxate/Meclosulfonate = 1/0.65-1/0.72; - Meclofenoxate/Eclofenoxate = 1/0.79-1/0.83; - Meclofenoxate/Adafenoxate = 1/0.20-1/0.26.

B) Examples 1-4 not being not limitedly, for producing the enterosoluble unit, oxopyrrolidine acetamides can be associated with hydrooxopyrimidine carboxylates. Their ratio is calculated per unit of anhydrous orotic acid, so that the association (wt/wt) between orotic acid and piracetam is in the weight ratio of 1/1.50-1/1.60, as can be seen from this example, which also carries out the therapeutic purposes of the drug. In this way, the composition for 100 enterosoluble units is as follows : EP 0 804 239 B1

- Orotic acid, anhydrous (1 ,2,3,6-Tetrahydro-2,6-dioxo-4-pyrimidinecarboxylic acid) 1 5.20 g -Piracetam (2-Oxo-1 -pyrrolidine acetamide) 24.00 g - di-Potassium hydrogen phosphate, anhydrous 8.80 g - Lithium carbonate 0.20 g - Nicotinic acid with prolonged-release 7.00 g - Magnesium oxide 1 .80 g -Potassium iodide 0.01 g -D(-) Fructose 0.90 g

Example 6

[0030]

A) Examples 1 -5 do not limit the invention, so that in manufacturing the gastrosoluble unit, aminoethanol phenoxyacetates can be replaced, as follows:

a) totally by aminoethyl phenoxyacetamides, like:

- 6.00 g - 8.00 g Mefexamide hydrochloride (N-[2-Di-ethylamino)ethyl]-2-(p-methoxyphenoxy)acetamide hydrochloride; - 1 .80 g - 2.20 g Fenoxedil hydrochloride (2-(p-Butoxyphenoxy)-N-(2,5-diethoxyphenyl)-N-[2-(diethylamino)ethyl]acetamide hydrochloride); - 2.50 g- 3.50 g Fexicaine hydrochloride (2-(p-Butoxyphenoxy)-N-(o-methoxyphenyl-N-[2-(1-pyrrolidi- nyl)ethyl]acetamide hydrochloride); - 6.00 g- 8.00 g Fipexide hydrochloride (1-[(p-Chlorophenoxy)acetyl]-4-piperonylpiperazine hydrochloride);

b) or partly by aminoethyl phenoxyacetamides, leading to the association between active substances from the two classes, which also achieves the therapeutic purposes of the drug. For illustration, the replacement of part of meclofenoxate hydrochloride by mefexamide hydrochloride is presented below.

Accordingly, the composition for 100 gastrosoluble units is the following:

- DL Methionine 16.50 g - Meclofenoxate hydrochloride (2-(Dimethylamino)ethyl(p-chlorophenoxy)acetate hydrochloride) 22.83 g - Mefexamide hydrochloride (N-[2-(Diethylamino)ethyl]-2-(p-methoxyphenoxy)acetamide hydrochloride) 6.52 g - Zinc sulfate, anhydrous 0.90 g - Nicotinic acid • Mg salt, anhydrous, with immediate-release 1 .00 g - L(+) Aspartic acid 9.20 g -D(-) Fructose 1.10 g EP 0 804 239 B1

(continued) - Vitamin B1 mononitrate 0.90 g -Vitamin B6 hydrochloride 1.20 g

The total or partial replacement of meclofenoxate by aminoethyl phenoxyacetamides is made within the range of the following ratios (wt/wt) between meclofenoxate and its substitutes:

- Meclofenoxate/Mefexamide = 1/0.24-1/0.29 10 - Meclofenoxate/Fenoxedil = 1/0.07-1/0.08 - Meclofenoxate/Fexicaine = 1/0.10-1/0.13 - Meclofenoxate/Fipexide = 1/0.24-1/0.29

B) The examples based on orotic acid are not restrictive, so that part of the quantity of orotic acid may be replaced, 15 within the same limits, with orotic acid salts of bioelements in the formulation, with or without crystallization water, as is described in this example; the therapeutic purposes of the drug are fulfilled also in this case. Then, the composition for 100 enterosoluble units is as follows:

- Orotic acid, anhydrous (1 ,2,3,6-Tetrahydro-2,6-dioxo-4-pyrimidinecarboxylic acid) 1 9.00 g -Orotic acid • Zn salt 1 .50 g - di-Potassium hydrogen phosphate, anhydrous 8.80 g - Orotic acid • K salt 4.50 g - Orotic acid • Li salt 1 .30 g - Nicotinic acid • Mg salt, anhydrous, with prolonged-release 9.00 g - Magnesium oxide 2.20 g - Orotic acid • Mg salt 9.70 g - Sodium iodide 0.014g -D(-) Fructose 1.10 g

A minimum active anti-stress daily dose consists in the administration of two gastrosoluble units and one enterosoluble unit. The pharmaceutical technology of the two complementary units develops according to the stages described in 40 example 1 .

Brief Description of the Drawings

[0031 ] The anti-stress, anti-impairment and anti-aging drug, according to the invention, was checked up on laboratory 45 animals. In the following are described three tests of preclinical verification, connected with an etio-pathogenic-therapeutic trivalent experimental pattern, shown in table 1 , which analyze normal (control, healthy) brains in comparison with those aggressed by stress and those therapeutically revitalized by the anti-stress, anti-impairment and anti-aging drug. The brain incapacitation was achieved by three types of stress, specific to humans (psychic, biological and global), and the anti-stress revitalization by the administration of the drug - prophylactic, therapeutic and restorative. so [0032] The aims of the complex experimental pattern, created by the authors of the drug, according to the invention, were to define structurally, ultrastructurally and biochemically the state of brain normality, to identify the characteristic damages induced in the brain through aggressing it by stress, wear and tear, and aging, to establish the main mechanisms of their occurrence and, proceeding from these facts, to elaborate, apply and preclinical^ check up an etio-pathogenic efficient therapy against stress, impairment and aging (RIGA, S., RIGA, D., Results of preclinical researches on 55 the experimental cheking-up of the Antagonic-Stress drug, Institute of Neurology and Psychiatry, Bucharest, 1985- 1989, in Romanian). [0033] For each kind of stress, the nervous system was investigated using three categories of methodology. The first consisted in the selective isolation from the brain of living cells (neurons and neuroglias) and their analysis by phase-

14 EP 0 804 239 B1

contrast microscopy (NORTON, W.T., PODUSLO, S.E., Neuronal soma and whole neuroglia of rat brain: a new isolation technique, Science, 167, 1144-1146, 1970; RIGA, D., RIGA, S., VARZARU, A., PERIETEANU, M., MARCULESCU, Z., Morphological studies on neuronal and glial live cell populations, selectively isolated from the brain. Light microscopy investigations (Part I), Military Sanitary Review, 87, 141-147, 1984, in Romanian). The second methodology was neu- 5 romorphology, the structural analysis of the brain being done by light, fluorescence microscopy (THOMPSON, S.W., HUNT, R.D., Selected Histochemical and Histopathological Methods, C.C. Thomas, Springfield, III., 1966) and electron microscopy (HAYAT, M.A., Principles and Techniques of Electron Microscopy, vol. 1 (Biological Applications), Van Nos- trand Reinhold, New York, 1970; RIGA, D., RIGA, S., POPESCU, A., PERIETEANU, M., CONSTANTINESCU, E., The methodology of lipofuscin pigment research in neuroanatomy, 3rd International Symposium on Teaching of Morpholog- 10 ical Sciences, Tel Aviv, Israel, 1977; Israel Journal of Medical Sciences, 14, 903, 1978). The third methodology, neurobiochemistry, was performed by isolation and assay of informational macromolecules from brain - nucleic acids (SANTEN, R.J., AGRANOFF, B.W., Studies on the estimation of deoxyribonucleic acid and ribonucleic acid in rat brain, Biochimica et Biophysica Acta, 72, 251-262, 1963; MUNRO, H.N., FLECK, A., Recent developments in the measurement of nucleic acids in biological materials, Analyst, 91, 78-88, 1966) and proteins (LOWRY, O.H., ROSENBROUGH, 15 N.J., FARR, A.L., RANDALL, R.J., Protein measurement with the Folin phenol reagent, Journal of Biological Chemistry, 193, 265-275, 1951; LAJTHA, A. (Ed.), Handbook of Neurochemistry, vol. I (Chemical Architecture of the Nervous Sys- tem), Plenum Press, New York, 1969). [0034] The purpose of using these methodologies, which reciprocally checked and confirmed their results, was completely to study the normal brain, the brain aggressed by stress and the brain therapeutically revitalized against stress, 20 wear and tear, by quantitative and qualitative methods at all its levels of morpho-biochemical organization (regional, tis- sual, cellular, subcellular and macromolecular levels).

Preclinical test I. The prophylactic efficiency of the drug, according to the invention, in protecting the brain from its aggression by psychic stress 25 [0035] Brain incapacitation by psychic stress consisted in the daily induction of chronic painful emotion to adult rats, for 2 weeks. A permanent 4 mA current supply to the cage floor produced the conditioned response of climbing up on a platform placed inside the cage; after 30 minutes, a 6 mA current was supplied to this platform for 2 seconds, at ran- dom intervals, for 3 hours, which caused the chronic painful emotion (adapted from DESIDERATO, O., TESA, M., 30 Shock-stress, gastric lesions and habituation in the chronic gastric fistula rat, Physiological Behaviour, 16, 67-73, 1976). In this way we created an animal pattern of experimental neurosis (stress, neurosis, painful emotion, fear, anxiety). The prophylactic protection of the brain was carried out by daily administration of the drug according to the invention, before aggression through psychic stress, during the same period of 2 weeks. [0036] As a methodology of investigation is exemplified by selective isolation of living cells (neurons and neuroglias) 35 from the cerebral cortex and their analysis by phase-contrast microscopy; thus, the morphological features of normality, the cellular damages determined by psychic stress and anti-stress therapeutic protection of nervous structures were established and compared. [0037] The preclinical checking-up of the prophylactic efficacy of the drug, according to the invention is presented in connection with figures 1 -6, which comparatively show the specific protection against psychic stress, attained at the cel- 40 lular level (soma and extensions) and the subcellular level (by stabilization of lysosomal membranes, protection of lipo- proteins membranes of other neuro- and glioplasmatic organelles and by the anti-catabolic effect, ultrastructural homeostatic effect - the preservation of subcellular integrity.

Preclinical test II. The therapeutic efficiency of the drug, according to the invention, in recovering the informa- 45 tional macromolecules from the brain, disturbed by biological stress aggression

[0038] Brain incapacitation by biological stress was fulfilled through its chronic intoxication with ethanol (in a concentration of 15 %, daily, per os, in drinking water) of adult rats, for 2 months. Thus, an animal pattern of chronic alcoholism has been created, using the digestive route. Brain revitalization was performed by daily administration of anti-stress so treatment with the drug, according to the invention, at the same time as the biological stress, and within the same period of two months. [0039] As a methodology of investigation it is exemplified by the isolation and assay of the informational macromolecules from the brain - nucleic acids (DNA and RNA) and proteins (total, water soluble and water insoluble); in this way, the neurobiochemical typology of normality, the disturbances of nervous macromolecules caused by biological stress 55 and the therapeutic normalization against stress were established and compared. [0040] The preclinical verification of the therapeutic efficacy of the drug, according to the invention is demonstrated by the data from tables 2 and 3. Comparing the results, the characteristic action against stress and impairment is evident, because the disturbances caused by biological stress at the level of informational macromolecules from the nerv-

15 EP 0 804 239 B1

ous system are diminished. The treatment engendered a marked stimulation of anabolism, ribosomal and proteinic regeneration, reflected by an increase in the amount of total RNA, total proteins (TP), water soluble proteins (WSP) and by the rise of RNA/DNA, RNA/TP ratios, accompanied by the anti-catabolic protection of the nerve cells, the protection from oxidative stress and from macromolecular cross-linkages, demonstrated by a decrease in the amount of water 5 insoluble proteins (WIP) and of (WIP/TP) • 1 00 ratio. The efficiency of revitalization treatment against stress, statistically significant, ranges between 64.7 % (for total DNA) and 85.2 % (for total RNA), as can be seen in table 2, and between 76.4 % (for WIP/TP • 100 ratio) and 94.0 % (for RNA/TP ratio), as shown in table 3.

Preclinical test III. The therapeutic restorative efficiency of the drug, according to the invention, in revitalization 10 of nervous structures and in recovering the informational macromolecules from the brain, disturbed by global stress aggression

[0041 ] Brain incapacitation by global stress consisted in accumulation of chronic wear and tear in adult rats (8-9 months) through their aging during 16-17 months up to senescence (24-26 months), concomitantly with their socio-sen- 15 sory deprivation, thus creating an animal pattern of accelerated impairment and aging. The brain revitalization was achieved by daily administration to old rats of the restorative treatment with anti-stress, anti-impairment and anti-aging actions, for 2 months, using the drug, according to the invention. [0042] The complex investigation of the brain is selectively exemplified by methodologies of neuromorphology (light, fluorescence and electron microscopy) and of neurobiochemistry (isolation and assay of the informational macromole- 20 cules). Their results were mutually verified and confirmed. The macromolecular-structural typology of the normal brain, of the brain incapacitated through global stress and revitalized by therapeutic recovery has been established and compared. [0043] The preclinical checking-up of the therapeutic restorative efficacy of the drug, according to the invention is shown in connection with figures 7-27 and tables 4 and 5. The results presented comparatively, demonstrated that the 25 drug had significantly reduced the disorders and lesions determined in the brain by stress, chronic wear and tear and accelerated aging, based on its intense and complex specific etio-pathogenic actions of neuropsychic and biological revitalization. [0044] The treatment efficiency on the catabolic regulation of nerve cells was demonstrated by the marked decrease of lipofuscin pigments, accompanied by the subcellular revitalization of neurons. These therapeutic effects are detected 30 at cellular-subcellular level by light microscopy-cytochemistry, in connection with figures 7-9, by fluorescence microscopy, connected with figures 10-12, and by electron microscopy, in connection with figures 13-15. The antagonization of the oxidative stress, checked up by the intense diminution of lipofuscin pigments, was fulfilled in conformity with a complex mechanism: anti-lipofuscinogenesis, neuronal and glial lipofuscinolysis and lipofuscin elimination, by the transport of the previously processed lipofuscin (fragmentation and partial solubilization) from neurons into glias and 35 then into capillaries. The successive stages of the mechanism, in their dynamics, have been demonstrated by electron microscopy of the brain, the initial and intermediate stages in connection with figures 16-18, and intermediate and final stages, connected with figures 19-21. [0045] The treatment efficiency on the anabolic regulation of nerve cells was demonstrated by repopulating them with Nissl bodies (agglomerations of rough endoplasmic reticulum and free ribosomes), accompanied by the regeneration 40 and revitalization of the other neuronal subcellular compartiments. The therapeutic effects were evinced at cellular-subcellular level by light microscopy-cytochemistry, in connection with figures 22-24, and by electron microscopy, connected with figures 25-27. The antagonization of the chronic stress and accelerated nervous impairment (hypoanabolism + hypercatabolism), checked up by their repopulation with Nissl bodies and subcellular neuronal regeneration has been carried out in conformity with a complex mechanism: anabolic, ribosomal and proteinic regen- 45 eration, emphasized by the quantity enhancement of total RNA, total proteins, water soluble proteins and of RNA/DNA, RNA/TP ratios, associated with the protection from catabolism and macromolecular cross-linkages, demonstrated by a decrease in the amount of water insoluble protein and in the (WIP/TP) • 1 00 ratio, as can be seen in tables 4 and 5. The efficacy of recovery and of anti-stress, anti-impairment and anti-aging revitalization, statistically significant, ranges between 45.0 % (for WIP) and 82.0 % (for total RNA), as shown in table 4, and between 55.9% (for (WIP/TP) • 100 so ratio), and 1 1 1 .3% (for RNA/TP ratio), as can be seen in table 5. [0046] At the three above described tests for preclinical checking-up of the drug, the results obtained were presented using only a part of methodologies:

the selective isolation from the brain of the living nerve cells at preclinical test I, 55 - neurobiochemistry at preclinical test II, neuromorphology and neurobiochemistry at preclinical test III.

By applying the other brain investigations, indicated in table 1 , to each type of stress, similar and equivalent results were

16 EP 0 804 239 B1

obtained, which are also significant (different only in their intensity); brain incapacitation by psychic, biological and global stress, and the therapeutic results have re-confirmed the treatment efficiency against stress, impairment and aging with the drug, according to the invention, both in prophylactic use and in therapeutic and restorative utilization. [0047] Using the experimental pattern of neuro-psycho-biology and the complex of methodologies, the authors of the 5 invention demonstrated etio-pathogenically that the three types of stress (psychic, biological and global) cause, accel- erate and intensify wear and tear and aging of the brain, by exceeding the nervous system adaptive capacities (includ- ing those against oxidation and against free radicals), owing to the intensity, frequency and chronicization of stress. In this way, the etiology, consisting in the aggression by stress and chronic impairment (ischemic-hypoxic stress, oxidative stress, free radicals, peroxidation) determines in the brain a characterisitic pathology, namely structural and biochemi- 10 cal disturbances and damages, at cellular, subcellular and macromolecular levels (peroxidation, polymerization, cross- linkages). The structural and biochemical diagnosis of this pathology was done by the demonstration of anabolism (regeneration) decreasing, catabolism (degradation) increasing and by the accumulation of lipofuscin pigments in the nerve cells. [0048] The efficiency of the anti-stress, anti-impairment and anti-aging etio-pathogenic actions preclinical^ tested of 15 the drug, according to the invention, is supported precisely by correcting the disturbances and antagonizing the lesions determined in the brain by stress, as it also can be seen from tables 6 and 7. In these tables, the stages and therapeutic mechanism in etio-pathogenic sequence, experimentally attested on animal patterns, at nervous cellular, subcellular and macromolecular level, are summarized. They emphasize the therapeutic intervention, metabolic and ultrastructural, in the two major directions of the metabolism, the anabolic regeneration and the catabolic regulation with the 20 removal of peroxidated subcellular wastes. These therapeutic mechanisms engender ultrastructural and metabolic homeostatic interventions and form the basis for nerve cell plasticity, for enhancing of nervous resistance and for the adaptive capacity of the body - the natural anti-stress processes. [0049] The experimental pattern described, based on the three examples of preclinical checking-up, further demon- strate the prophylactic, therapeutic and restorative efficiency of the drug, according to the invention, against stress, 25 impairment and aging, precisely in those zones of the brain which are directly involved and continously overstrained by the aggression and chronicization of stressors, by the permanent effort of the general adaptation reaction and by the neuropsychic and biological impairment. As can be seen from the figures and tables, this structural-macromolecular specificity of the therapeutic action is represented in the brain as follows:

30 - at the regional level: telencephalon, cerebral cortex, hippocampus; diencephalon; reticular formation; at the tissue and cell level: neurons and neuroglias (astrocytes, microglias and oligodendrocytes); at the subcellular level: the anabolic system (Nissl bodies; free ribosomes, rough endoplasmic reticulum, Golgi apparatus) and the catabolic system (lipofuscin pigments; primary --> secondary --> tertiary lysosomes); at the macromolecular level: nucleic acids (DNA and RNA) and proteins (total, soluble and insoluble). 35 [0050] As part of preclinical verification, complementary researches on guinea pigs and mice were performed, comparatively demonstrating that the aggression by stressors-impairment-aging causes in the long run, the same damage typology (metabolic and ultrastructural) in rat, guinea pig and mouse brain, irrespective of the animal species. Our results are also supported by similar data reported in the prior art for rodents (SAMORAJSKI, T, KEEFE, J.R., ORDY, 40 J.M., Intracellular localization of lipofuscin age pigments in the nervous system, Journal of Gerontology, 19, 262-276, 1964; JOHNSON, Jr. J.E., MIQUEL, J., Fine structural changes in the lateral vestibular nucleus of aging rats, Mecha- nisms of Ageing and Development, 3, 203-224, 1974), primates (BRODY, H., HARMAN , D., ORDY, J.M. (Eds.), Aging, vol. 1 (Clinical, Morphologic, and Neurochemical Aspects in the Aging Central Nervous System), Raven Press, New York, 1975) and humans (BRODY, H., The deposition of aging pigment in the human cerebral cortex, Journal of Geron- 45 tology, 15,258-261, 1960; RIGA, D., Light and electron microscopic topography of changes induced by aging in the central nervous system, Annual scientific session of "Victor Babes" Institute, Bucharest, 29 March, 1974, in Romanian; DAVISON, A.N., THOMPSON, R.H.S. (Eds.), The Molecular Basis of Neuropathology, Edward Arnold, London, 1981; CONDORELLI, D.F, AVOLA, R., RAGUSA, N., GIUFFRIDA STELLA, A.M., Macromolecular synthesis in developing and aging brain, pp. 243-257, In: CASTELLANI, A., BALDUINI, C, VOLPE, P. (Eds.), Macromolecules in the Function- so ing Cell, Consiglio Nazionale delle Ricerche, Roma, 1988). [0051] Therefore, the same lesional typology (animal-human) and the fact that we incapacitated the animal brain using types of stress that are characteristic of man (psychic stress - painful emotion, anxiety; biological stress - ethanol dependence; and global stress - accelerated wear and tear and aging) lead to the conclusion that the actions and mechanisms of anti-stress revitalization (the preclinical checked up on animal brain) are similarly achieved in human 55 brain. [0052] This therapeutic conclusion is further demonstrated in three trials of clinical application of the drug according to the invention, using a clinical pattern approved by the Pharmacovigilance and Drug Commitee of the Ministry of Health in Bucharest, Romania. As pathology directly connected with stress, accelerated impairment and premature

17 EP 0 804 239 B1 aging, and for establishing an equivalence between the preclinic (animal) and clinic (human) pathologies and therapeutic actions, the clinical studies involved three diseases: neurasthenia, chronic alcoholism and chronic cerebral circulatory insufficiency.

Clinic trial I. The therapeutic efficiency of the drug, according to the invention, in the treatment of neurasthenia

[0053] Neurasthenia (fatigue syndrome), classified by WHO in the category of neurotic, stress-dependent and somatoform disorders, was studied as neurasthenia disease (20 patients) and syndrome (10 patients), in order to test the drug over a wide rage of neurastheniform, psychosomatic and somatopsychic disorders. [0054] The characteristics of the group investigated were: 30 patients (15 hospitalized and 15 in activity, meaning in conditions of chronic stress), of both sexes (15 males and 15 females), aged between 20 and 58 (mean age - 37 years). The selected group was homogeneous, thanks to the application of criteria for inclusion (positive diagnosis), connected with criteria for exclusion (differential diagnosis), (World Health Organization (WHO), Tenth Revision of the International Classification of Diseases (ICD-10), Chapter V (F): Mental, Behavioural and Developmental Disorders, WHO, Geneva, 1987), and with registering the similar symptomatologic intensities and performance deficiencies. The treatment with the anti-stress, anti-impairment and anti-aging drug was exclusive, in oral posology, for one month. [0055] The assessment methodology, administered before, during and at the end of the treatment period, comprised: a scale for neurasthenia evaluation (having 10 target symptoms, each of them with 4 intensity degrees), 8 psychological tests (7 psychometric for attention, memory, general intellectual operativity, total average cognitive performance and one personality test), paraclinical investigation - bioelectric (EEG and ECG recordings) and biological-humoral (usual laboratory tests). [0056] The therapeutic efficacy of the drug, according to the invention was very good and good, in both the neurasthenia disease and syndrome, as well as upon the neurasthenic in-patients and those still on their jobs; in the meantime the neuropsychic and somatic effects were maintained over the long term. No adverse reactions were registered. The therapeutic benefit obtained after 1 -2 weeks of treatment (GORGOS, C, BOTEZAT-ANTONESCU, I., Results of clinical trial with Antagonic-Stress drug in neurasthenia, "Titan" University Polyclinic, "Titan" Mental Health Centre, Bucharest, 1988, in Romanian) was objectified clinically and paraclinically by:

a) quantitative and qualitative regression and the disappearance of the symptomatology specific to neurasthenia, the normalization of the wakefulness-sleep biorhythm, the disappearance of neurovegetative troubles and of psychosomatic disorders; alongside with b) normalization of performances in the concrete activity, with the recovery of motivation and work capacity, accompanied by the increase of professional efficiency; c) emotional stabilization (overall affective) and enhancement of the neuropsychic resistance to aggression by psychosocial stressors in case of continuing work in a stress-inducing environment; d) regulation of cerebral electrogenesis, as a consequence of metabolic regulation and functional normalization of the nerve cells; e) emphasizing the myocardial anti-ischemic, moderate hypotensive, hepatoprotective and normolipidemic effects in those patients who suffered from these disorders before treatment.

Clinical trial II. The therapeutic efficiency of the drug, according to the invention, in the treatment of chronic alcoholism

[0057] Chronic alcoholism (ethanol dependence) was moderate and severe, the duration of addiction between 1 0 and 35 years, with neuropsychiatric and somatic impairment and negative psychosocial consequences. [0058] The characteristics of the group investigated were: 30 in-patients, only males (since the frequency of heavy drinkers is 2-5 times higher in males than in females), aged between 31 and 59 (mean age - 47 years). The homoge- neity of this selected group was obtained by applying the criteria for inclusion and inside each of them the criteria for severity (anamnestic, psychiatric-psychological, psychosocial, somatic, screening tests, lack of therapeutic results through previous complex drugs and treatments), as well as of criteria for exclusion (American Psychiatric Association, Diagnostic and Statistical Manual of Mental Disorders (DSM-III-R), third edition, revised, American Psychiatric Associ- ation, Washington, DC, 1987). The treatment with the anti-stress, anti-impairment and anti-aging drug was exclusive, in oral posology, for one month. [0059] The assessment methodology, administered before, during and at the end of the treatment period, involved: psychiatric evaluation - interview-examination, the scale for chronic alcoholism evaluation (10 items), Hamilton depression scale (21 items); psychologic assessment - Bourdon-Anfimov, Herwig III, subtest COD from Wechsler Adult Intel- ligence Scale, Klazow, auditive Ray, Wechsler memory scale (M.Q.), Raven (I.Q.-matrix 1938, I list), Luscher; clinical evaluation - neurologic, ophthalmologic, of internal medicine; paraclinical assessment - bioelectric (EEG, ECG record-

18 EP 0 804 239 B1

ings), hematologic, metabolic, immunologic and screening tests. [0060] The evident therapeutic value consisted in obtaining a rapid therapeutic benefit, which was constant, intense, multiple and persistent. No toxic effects and adverse reactions were noticed. The therapeutic benefit (PREDESCU, V, NEICU, N., STEFANESCU, P., PRELIPCEANU, D., ANDRUCOVICI, I., DAVID, E., VRABIE, E., ROMAN, I., SIMOCA, 5 I., BUDILEANU, M., SIMA, I., NEGRU, T, DUMITRESCU, G., REU, G., Results of clinical trial with Antagonic-Stress drug in psychic impairments induced by constant alcohol consumption (chronic alcoholism, ethanol dependence), "Gh. Marinescu" Clinical Hospital, Department of Psychiatry, Bucharest, 1988, in Romanian), objectified through a complex clinical and paraclinical methodology, is the consequence of multiple actions of the drug and its effects:

10 a) psychotropic consequences - psychotonic of metabolic regulation, antidepressive, nootropic, sleep normalization, neuropsychic revitalization; remission, under treatment, of the withdrawal symptomatology, of the acquired dependence and tolerance; synergistic effect by increasing (normalization) and supporting the cognitive performances, concomitantly with the behaviour stabilization, in point of affective/emotional and relational/social, which enhance the resistance to effort and stressor aggression; 15 b) neurotropic effects - cerebral vasodilative, correction of the cerebral electrogenesis, regression up to disappearance of the signs and symptoms of alcohol toxico-def iciency encephalo-myelo-polyneuropathy; c) somatotropic consequences - ocular - trophic and vasodilative action; hepatoprotective and anti-toxic - fast remission of the hepatic cytolytic and insufficiency syndroms; cardiovascular - improvement of myocardial irriga- tion, decrease of bioelectric signs from alcoholic myocardiopathy, disease also favorized by chronic cigarette smok- 20 ing, moderate hypotensive effect; the general revitalization of the organism and anabolic stimulation; d) hematologic effects - anti-anemic action, normalization of macrocytosis and of mild leukocytosis; e) metabolic consequences - anabolic regeneration, hypolipidemic and hypoglicemic actions; f) immunologic effects - immunostimulation by recovery of the humoral and cell-mediated immune deficiency.

25 Clinical trial III. The therapeutic efficiency of the drug, according to the invention, in the treatment of chronic cerebral circulatory insufficiency

[0061] Out of the types of chronic cerebral circulatory insufficiency we chose the decompensations occurring in the vertebrobasilar territory, the patients having at the same time a fragile and deficient vascular system, a somatic back- 30 ground of biological impairment and aging and an organism that had been marked by stressors, risk factors and stress- related illnesses. [0062] The characteristics of the group investigated were: 30 in-patients, of both sexes (1 1 males and 19 females), aged between 47 and 72 (mean age - 59 years). The selected group was homogeneous thanks to the application of criteria for inclusion (positive diagnosis) and criteria for exclusion (differential diagnosis), (Organisation Mondiale de la 35 Sante (OMS), Application a la Neurologie de la Classification Internationale des Maladies (CIM-AN), OMS, Geneve, 1989, in French). The treatment with the anti-stress, anti-impairment and anti-aging drug was exclusive, in oral posology, and lasted one month. [0063] The assessment methodology, administered before, during and at the end of treatment period, involved: the neurologic examination card, the evaluation scale of chronic cerebral circulatory insufficiency (10 target symptoms, 40 each of them with 4 intensity degrees), bioelectrical recordings (EEG, ECG), and biological-humoral investigation (laboratory tests). [0064] The therapeutic efficacy of the drug, according to the invention, was very good (rapid, global and significant). No adverse reactions were registered. The therapeutic benefit (STAMATOIU, I., DIMITRIU, R., NICOLAE, I., Results of clinical trial with Antagonic-Stress drug in chronic cerebral circulatory insufficiency from vertebrobasilar territory, Clini- 45 cal Hospital of Bucharest Municipality, Department of Neurology, Bucharest, 1988, in Romanian; POPA, C, MACOVEI, M., Results of clinical trial with Antagonic-Stress drug in chronic vertebrobasilar insufficiency, "Gh. Marinescu" Clinical Hospital, Department of Neurology, Bucharest, 1988, in Romanian), clinically and paraclinically objectified, confirmed the efficiency of the drug in cerebroprotection and in the treatment of brain aggression by ischemia, hypoxia, chronic stress and accelerated wear and tear, and consisted of: 50 a) the rapid regression of the neurologic and asthenic symptomatology, accompanied by the disappearance of symptoms; b) the obvious improvement of cortical electrogenesis and the decrease down to disappearance of the bioelectric sign of myocardial ischemia; 55 c) obtaining a global biological response of neuropsychic revitalization (through cerebral vasodilative, nootropic and psychotonic metabolic effects) and somatic revitalization (through hepatoprotective, moderate antihyperten- sive and normolipidemic consequences, as well as by the synergism with hypoglycemic medication).

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[0065] The versatile and superior therapeutic efficiency of the drug, according to the invention, emphasized by the three preclinical tests and the three clinical trials, was obtained thanks to the fact that the drug was developed in terms of a biological drug, as can be seen from the pharmaceutical examples, and due to its specific and synergistic anti- stress, anti-impairment and anti-aging actions. [0066] Thus, the drug is a synergistic biological, neurometabolic and cell-trophic composition with active anti-stress principles (psychotropic, neurotropic, hepatotropic and myocardiotropic constituents), basic (normal) components of the biological systems and of the central nervous system, which pass through the blood-brain barrier, are actively and physiologically integrated in the biochemistry of neurons and neuroglias, specifically control the cerebral circulation and metabolisms. Being a biological drug, it is well accepted by the organism and is non-toxic, it produces no adverse reactions, tolerance and dependence, and it exhibits no danger of overdosage or intoxication; the drug itself has anti-toxic, detoxicant and homeostatic effects. Furthermore, the possibility appears to employ it in a long-term administration, as a drug for neuropsychic and biological support, for obtaining and consolidating the actions of vascular, energetic, polymetabolic and ultrastructural regulation of the brain, which are necessary in the conditions of magnifying and chronicization of stressors and intensification of neuropsychic-biological wear and tear. [0067] The specific homeostatic and etio-pathogenic actions of the drug against stress, wear and tear and aging are achieved directly (primarily and secondarily) or as pro-drugs, through its active anti-stress components, with synergistic and polyvalent actions:

a) against oxidative and catabolic stress; b) against anabolic stress; c) vasodilative and normolipidemic; d) energo-active; and e) anti-toxic.

a) The components active against oxidative and catabolic stress of the anti-stress, anti-impairment and anti- aging drug, according to the invention, are: methionine, aminoethanol phenoxyacetates (meclofenoxate, meclosulfonate, eclofenoxate, adafenoxate) and/or aminoethyl phenoxyacetamides (mefexamide, fenoxedil, fexicaine, fipexide). Methionine is a natural antioxidant, which antagonizes oxidative stress, through the biosynthesis of thiolic compounds, as well as catabolic stress, being a donor of methyl groups, metabolically sustaining against stress these biochemical reactions of the general adaptation syndrome. Meclofenoxate is a biological compound, an organic ester, which is hydrolyzed in brain into two natural metabolic active substances: p-chlorophenoxyacetic acid, a phytohormone (auxin) and a cerebral modulator of neurotransmitters, and dimethylaminoethanol, a natural aminoalcohol, a precursor of brain acetylcholine and of cerebral lipids (phospholipids and sphingolipids). The etio-pathogenic action of the active components against oxidative and catabolic stress is achieved by ensuring the metabolic support: the sustenance of anti-oxidant defence biological systems, the free radical scavenging ability, the antagonization of lipid peroxidation, polymerization and cross-linkages in macromolecules (antagonization the mechanisms of lipofuscin pigment formation and accumulation), lipofuscinolysis, re- balancing of the negative balances (thiolic, sulfur, methyl, choline, phosphatides) caused by stress, aging, hypoxia-ischemia and alcoholism in the organism and brain, the antagonization of excessive calcium accumulation in the brain (induced by the damage of cell membranes and free radical attack). In this way, the active principles against oxidative and catabolic stress effect also an anti-toxic action in the nervous system. The specific action of constituents against oxidative and catabolic stress is amplified by the contribution of the other active principles of the drug according to the invention. Thus, the compounds against anabolic stress act through orotic acid (lipofuscinolysis), piracetam (decreasing the quantity of cerebral nonesterified fatty acids, from the increased amount due to stress), zinc (anti-oxidant), lithium (anti-catabolic), whereas the whole class operates through anabolic regeneration, which opposes catabolism, and degradation. The vasodilative and normolipidemic components act through nicotinamide (which diminishes the cerebral nonesterified fatty acids, existing in increased amount due to stress) and through the antagonization of hypoxic-ischemic stress and of chronic alcoholism, which cause lipofuscin pigment accumulation in the brain. In their turn, the anti-toxic constituents support the homeostatic action of the active principles against oxidative and catabolic stress, by detoxifying and elimination of the catabolism products at structural, metabolic and molecular level in the nervous tissue. Thus, the active principles against oxidative and catabolic stress are geroprotective agents, because they decelerate subcellular wear and tear and aging processes in the brain, which is the tissue having increased fragility to oxidative stress and free radical attack.

20 EP 0 804 239 B1

The therapeutic efficiency of the drug according to the invention in antagonization of the oxidative and catabolic stress was demonstrated at the nervous level: by protection, stabilization and increasing the resistance of neuronal and glial membranes (in animal psychic stress); neurobiochemically by protecting cerebral macromolecules and diminishing the number of cross-linkages in protein macromolecules; also neuromorphologically (by light, fluorescence and electron microscopy), based on the action against lipofuscin formation, neuronal and glial lipofuscinolysis and that of lipofuscin elimination (in animal chronic alcoholism and aging). The homeostatic action against oxidative and catabolic stress was clinically objectified in the treatment of chronic alcoholism, by the remission of the hepatic and pancreatic cytolytic (hypercatabolic) syndrome, and by the rapid, functional and metabolic normalization of the liver. b) The components active against anabolic stress of the anti-stress, anti-impairment and anti-aging drug, according to the invention, are: hydrooxopyrimidine carboxylates (orotic acid and orotates of the drug bioelements) and/or oxopyrrolidine acetamides (piracetam, oxiracetam etiracetam, dimethylphenyl piracetam, pramiracetam, aniracetam) associated with potassium, zinc and lithium. Orotic acid (vitamin B13) has an intense action of anabolic regeneration, because it is the natural metabolic precursor common for pyrimidine nitrogenous bases (uracil, cytosine, thymine, 5-methylcytosine) from nucleic acids (DNA, RNA) and macro-ergic pyrimidine nucleotides (UTR CTR TTP), or from the composition of coenzymes (UDPglucose, UDPgalactose, CDPcholine), which interfere with carbohydrate and lipid metabolisms: these, alongside protein and nucleic acid metabolisms, are extremely important in the brain. Thus, orotic acid has a lipotropic effect, stimulating the phospholipid synthesis and also increasing the glycogen synthesis in hepatic, myocardial and nerve cells, and through enhancing RNA synthesis in brain it provides the subcellular and metabolic support for the long-term memory. Therefore, orotic acid represents a strong natural anabolic and biological support for the brain, liver and myocardium. Piracetam activates the biosynthesis of cerebral macromolecules (RNA, proteins) and lipids, especially in post-hypoxic recovery and in old animals, where it promotes polyribosome synthesis and their extension, by providing the energy transfer between the systems which produce energy and those which use it ( cellular anabolism, subcellular regeneration). Potassium, zinc, lithium and the other neuroactive bioelements of the drug according to the invention (magnesium, iodine, monoacid phosphate, sulfate) are intracellular cations and anions, mono- and divalent, essential for living matter. They play structural (plastic) and metabolic (dynamic) roles, and are involved in fundamental biocatalytic systems for the functional adaptation of the cell - enzymes, hormones, vitamins and macro-ergic compounds. They acquire specific functions in nerve cell metabolism and physiology: in neurotransmitter activity, in excitation-inhibition processes, in nerve conduction and axoplasmic transport, in memory and neurosecretion. They exhibit a negative balance in stress-impairment, chronic alcoholism, hypoxia- ischemia, especially magnesium, zinc, sulfate and iodine. Magnesium is a subcellular stabilizer, lithium a humoral stabilizer, and the limbic system displays increased sensitivity to this bioelement, while zinc has an increased concentration in hippocampus. The therapeutic efficiency of active compunds against anabolic stress is sustained also by other active principles of the drug, according to the invention, as already mentioned above for the other bioelements, taking into account the dependence of cerebral anabolism on the cerebral blood flow and on the nervous energy metabolism. Thus, the components against oxidative and catabolic stress act through methionine and aminoethanol phenoxyacetates. In the brain, the methyl groups supplied by methionine are intensely consumed in the anabolism processes of neurotransmitters, in cerebral lipid biosynthesis and in the post-transcriptional methylation of the nervous macromolecules - nucleic acids and proteins. Meclofenoxate promotes the synthesis of cerebral RNA and macro-ergic nucleotides, and, through its hydrolysis compound (dimethylaminoethanol), it contrib- utes in the same way as meclosulfonate, to the cerebral phospholipid synthesis. The active vasodilative components (against hypoxic-ischemic stress) provide the contribution of nutrients, which are necessary to anabolic regeneration and to antagonization of anabolic stress-inducing processes (hypoxic-ischemic stress). The energo-active compounds: aspartate (precursor in the biosynthesis of purines and pyrimidines), fructose, vitamin vitamin B6 and monoacid phosphate create the energetic support necessary for intensifying anabolism. The therapeutic efficacy of the drug according to the invention in antagonizing anabolic stress, in chronic alcoholism and nervous wear and tear, and the intense functional and ultrastructural anabolic activation and regeneration of nerve cells were demonstrated on animal brain, neurobiochemically, by enhancing the amounts of RNA, total proteins and water soluble proteins, as well as neuromorphologically (light and electron microscopy), by neuronal repopulation with Nissl bodies, rough endoplasmic reticulum and free ribosomes. Anabolic regeneration was clinically emphasized in the treatment of chronic alcoholism by remission of hepatic

21 EP 0 804 239 B1

insufficiency and immune deficiency, by the anti-anemic effect and weight regain. c) The active vasodilative (against hypoxic-ischemic stress) and normolipidemic components of the anti-stress, anti-impairment and anti-aging drug, according to the invention, are: nicotinic alcohol and/or acid or its derivatives (nicotinamide, magnesium nicotinate) associated with magnesium and iodine. They operate etio-pathogenically by increasing the cerebral blood flow, enhancing the permeability of the blood-brain barrier and the transport of glucose, by improving the nerve and myocardial cell supply with anabo- lites and their purification of catabolites. These compounds produce an antispastic, spasmolytic action, physiologic cerebral vasodilatation (primary - vasculotropic) and a natural anti-hypoxic and anti-ischemic protection of the brain. Furthermore, they act as normolipidemic and lipid homeostatic agents, by decreasing the high levels of triglycerides, total cholesterol, VLDL cholesterol (very low-density lipoprotein cholesterol), LDL cholesterol (low-density lipoprotein cholesterol), and by increasing the low levels of HDL cholesterol (high-density lipoprotein cholesterol). In its turn, the normolipidemic action exerts positive effects on cerebral vasodilatation. Moreover, the therapeutic efficiency of vasodilative and normolipidemic components is also based on the action of the other active substances of the drug, according to the invention. The active principles against oxidative and catabolic stress, those against anabolic stress and the energo-active ones operate as psychotropic- psychotonic compounds for metabolic regulation, and thus they produce a secondary cerebral vasodilatation - a neuromerometabolic one, which is subsequent to the functional and metabolic cerebral stimulation. The active components against oxidative and catabolic stress, orotates and potassium belonging to constituents against anabolic stress, and aspartates, fructose, vitamin B-|, vitamin B6 belonging to energo-active principles, antagonize the hypoxic-ischemic stress. Orotic acid, meclofenoxate, meclosulfonate and methionine act as blood normolipidemic agents, also through intensifying the intracellular lipid synthesis at hepatic, cerebral and myocardial levels or due to a choleretic effect (by magnesium and orotic acid). The vasoactive components of the drug, according to the invention, without retarded delivery in the gastrosoluble unit and with prolonged delivery in the enterosoluble one, provide both a rapid vasodilatation and one prolonged, over a long period (about 2-3 hours in case of the prolonged-release granule, and about 8 hours in case of the prolonged-release tablet), which are necessary in the control of hypoxia-ischemia induced by stress, impairment and normal and pathological aging. The concomitant administration of the two complementary units achieves sustained vasodilatation (immediate + prolonged), necessary in cerebral and myocardial ischemic pathology. The vasodilative action of the drug according to the invention may be enhanced by replacing, in the formulation of gastrosoluble unit, aminoethanol phenoxyacetates (base esters) by aminoethyl phenoxyacetamides (stabler amides), and among them, fenoxedil, a cerebral and peripheric vasodilator (based on vasculotropic- musculotropic mechanism) and psychotonic for metabolic regulation (vasodilatation through neurometabolic activation) is preferred. The therapeutic efficiency of the drug according to the invention in antagonizing hypoxic-ischemic stress and in dyslipidemias was clinically emphasized in the treatment of chronic cerebral circulatory insufficiency, by the remission of ischemic symptomatology, by ophthalmologic positive effects, by cerebral and myocardial electrogenesis correction, and in the treatment of chronic alcoholism by the normalization of the lipid profile and ocular positive effects. d) The energo-active components of the anti-stress, anti-impairment and anti-aging drug, according to the invention, are: aspartate, D(-) fructose, vitamin B-|, vitamin B6, monoacid phosphate, which support the energy metabolism of the nerve cell, overstrained and energetically exhausted during stress and impairment. Aspartic acid - a gluco-formative amino acid, in a 96 nmol/100 g nervous tissue concentration, stored in the brain in its deposit form, namely N-acetylaspartic acid in a 490 nmol/100 g nervous tissue concentration - supplies energy by its metabolization in tricarboxylic acid cycle and GABA shunt (extremely intense in the brain); it is also an excitatory neurotransmitter and achieves a mobile connection between carbohydrate, energy, nucleic acid and protein metabolisms from the nerve cell. D(-) fructose, a natural monosaccharide, which is metabolized independently of the insulin pathway, enhances the hepatic glycogen reserve or is rapidly integrated in the form of monophosphoric ester (D(-)fructose-6-phosphate) in the pentose phosphate shunt (oxidative degradation used by the nerve cells) or in the anaerobic glycolysis. Vitamin B-|, vitamin B6 and vitamin PP (nicotinamide), the last belonging to the vasodilative component, are exogenous biocatalysts, absolutely necessary for life and for the normal unfolding of the nervous actions. In their form of phosphoric acid esters (thiamine pyrophosphate and pyridoxal 5-phosphate) or of nucleotides (nicotinamide mono- or dinucleotides), they represent the coenzymes (prosthetic group) for numerous essential enzymes, which interfere in energy, carbohydrate, protein and lipid metabolisms. Because of the increased intensity of cerebral metabolisms, the nervous system concentrates these vitamin in different zones and it is sensitive to their imbalances: negative balance determined by stress, wear and tear, aging and toxico-defi-

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ciency induced by chronic alcoholism. Vitamin B-, is essential to the metabolization of lactic acid, pyruvic acid and in tricarboxylic acid cycle, and vitamin B6 to activating the GABA shunt, thus being nervous metabolic pathways to stress antagonization. The monoacid phosphate represents the support, a metabolic reserve in energy metabolism, and it is employed in glucide phosphorylation - for their entering in carbohydrate metabolism; in vitamins B-|, B6, PP phosphorylation - in order to activate them as coenzymes; and for the synthesis of macro-ergic compounds - ATP, UTP etc. - for energy storage. The therapeutic efficacy of the energo-active components is sustained by the action of other active principles of the drug, according to the invention, which interfere homeostatically in the main stages of the cerebral carbohydrate and energy metabolisms. Thus, the constituents active against oxidative and catabolic stress act through aminoethanol phenoxyacetates, which enhance the glucose and phosphate transfer from blood into brain, the oxidative degradation of glucose through the pentose phosphate shunt, the strong exergonic cycle of tricarboxylic acids, the oxygen utilization by the nerve cell; they also act through the aminoethyl phenoxyacetamides that achieve a strong energo-active, psychoenergizing action; through methionine they act indirectly, contributing to biological methylation (synthesis of adrenaline, acetylcholine and creatine). The compounds active against anabolic stress operate through orotic acid, which saves energy during the direct synthesis of pyrimidine nitrogenous bases, of the coenzymes and of macro-ergic compounds with uracil, during the re-synthesis of glycogen stocks and by piracetam, which stimulates the energy producing systems, increases the glucose consumption, the aerobic glycolysis, the ATP synthesis at the brain level and the "in vitro" respiratory coefficient of the mitochondrion (the subcellular organelle responsible for ATP biosynthesis); and through potassium and zinc, which interfere in many enzymatic reactions accompanied by release of energy. The vasodilative components (against hypoxic-ischemic stress) act by the enhancement of the anabolite and oxygen amounts in the brain; through nicotinamide, a precursor in the NAD and NADP coenzyme synthesis, playing an important role in the cellular oxidation-reduction processes, through magnesium which interferes directly in many exergonic reactions, in the biosynthesis of macro-ergic compounds, in oxidative phosphorylation; and through iodine which produces a general metabolic activation, mainly in the brain, and which is particularly high in the hypothalamus and also accumulates in the spinal cord and medulla oblongata. The association of energo-active neurometabolic components with those for nootropic-psychotonic of metabolic regulation (aminoethanol phenoxyacetates, aminoethyl phenoxyacetamides, hydrooxopyrimidine car- boxilates and/or oxopyrrolidine acetamides), with methyl group donors (methionine, pramiracetam) and with physiologic cerebral vasodilative substances (having a rapid and prolonged action) psychically creates a psychoenergizing (anti-asthenic) effect, the promotion of cognitive performances (attention, memory, intellectual promptness), an antidepressant effect, higher resistance to stress, wear and tear, hypoxia, and the normalization of cerebral electrogenesis. These effects have been clinically proved in the treatment of neurasthenia, chronic alcoholism and chronic cerebral circulatory insufficiency, by using the drug, according to the invention, e) The anti-toxic components of the anti-stress, anti-impairment and anti-aging drug, according to the invention, operate etio-pathogenically against hypercatabolic and toxic and deficient states, connected with the oxidative stress and the accumulation of endogenous toxic compounds, induced at ultrastructural and metabolic level by stress and impairment, hypoxia-ischemia, chronic alcoholism and aging. In this way the metabolic detoxifying processes are activated, which in their turn:

diminish the number of the free radicals generated by oxidative stress through aminoethanol phenoxyacetates, methionine, orotic acid and zinc; remove acidosis and cellular hypoxidosis, caused by the accumulation of lactic and pyruvic acids conse- quent upon oxidative stress, glucide hypercatabolism, thiamine-carbohydrate imbalance, hypoxia- ischemia, alcoholism through aminoethanol phenoxyacetates, orotic acid, potassium, zinc, magnesium, vitamin B-| and vitamin B6; remove ammonia generated in excess through protein catabolism by aspartic acid - through the urea cycle, by transamination - together with vitamin B6, asparagine and magnesium; intervene in sulfoconjugation by methionine and sulfate, in glucurono-conjugation by fructose, in methylation by methionine and pramiracetam; antagonize the excessive accumulation of calcium in the brain by aminoethanol phenoxyacetates.

The subcellular detoxifying processes are also activated, and they:

diminish the number of cross-linkages which make proteins-enzymes insoluble and inactive;

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decrease the lipid peroxidation determined by oxidative and catabolic stress, acting against lipofuscinogenesis, by lipofuscinolysis and lipofuscin elimination.

Methionine, an essential amino acid, a natural detoxicant and lipotropic, a source for sulfur, a metabolic donor of methyl groups, and a precursor of -SH and sulfate groups, plays a complex role in the anti-toxic action, thus being an important metabolic support for the anti-toxic and detoxifying activity. The oral administration of the drug, according to the invention offers the advantages of direct anti-toxic and hepatoprotective actions and of actively sustaining the hepatic metabolic detoxifying processes, taking into consideration the central role played in these processes by the liver. The fact has been clinically proved in the treatment of chronic alcoholism by using this drug. The cumulation of anti-toxic (metabolic - ultrastructural), vasodilative (anti-hypoxic, anti-ischemic, spasmolytic) and anabolic regeneration (subcellular - functional) actions with those against oxidative hypercatabolic stress (removal of neuronal lipid peroxides) and energo-active actions leads, at the plastic and functional level, to the trophic action (nervous, hepatic, myocardial) and to the neuropsychic and biological revitalization of the drug, according to the present invention.

Industrial Applicability

[0068] The therapeutic indications of the drug, according to the invention result from:

its integration into the fundamental processes of neurons and neuroglias and their support based on the biological, neurometabolic and cell-trophic composition; maintaining and increasing of the adaptive capacity and the neuropsychic and biological resistance by polymetabolic regulation and subcellular regeneration, first of all for nerve, hepatic and myocardial cells; the simultaneous achievement of brain, liver and heart protection from the incapacitation caused by acute and chronic stress, from cerebral impairment and the acceleration of aging, by homeostatic intervention at brain and body level; the correction of the main metabolic and ultrastructural imbalances brought about by stress, prolonged biological wear and tear and stress-related illnesses, thanks to its vasoactive, energo-active, anti-toxic actions and influences on catabolic regulation and anabolic regeneration; the complex, etio-pathogenic, anti-stress, anti-impairment, anti-ischemic, anti-hypoxic and anti-toxic involvement of the drug, based on its synergistic biological-metabolic composition (nootropics-psychotonics, vasodilators, ener- goactivators, stimulants of protein and nucleic acid synthesis, amino acids, vitamins and bioelements).

[0069] The administration of the drug, according to the present invention, to healthy humans (in harmony with the health promotion conception - preventive medicine) has the following effects:

it elevates the general vitality of the organism, consolidates the neuropsychic tonus; it enhances the capacity for psychic and physical efforts, for the intellectual and sportive efficiency and performances; it increases the neuropsychic and biological adaptive capacity; it expands the resistance to overstrain, stress and impairment; it determines the fast recovery in cases of fatigue, exhaustion, overwork, after prolonged efforts or sleeplessness; it strongly decelerates brain and body wear and tear and promotes the extension of the functional life span, and of longevity, thus preventing impairment, involution, presclerosis and atherosclerosis; its suited for an extensive use, for the whole range of ages (children, young people, adults, old people), being especially indicated for people over 35-40 (when the adaptive capacity and resistance start decreasing progressively and continuously) and more specifically in performant activities or those having a high stressing and aggressive coefficient (management personnel, students, sportsmen, pilots, teams of space flights and missions etc.).

[0070] Since brain stimulation is achieved by ultrastructural, energetic and metabolic regulation, the administration of the drug according to the invention is not accompanied by depression, nervous exhaustion, tolerance, psychological and physical dependence; on the contrary an intense, strengthening, neuropsychic and general effect is obtained for over the long term. [0071] The drug, according to the invention, is administered to sick persons for therapeutic and recovery purposes, increasing the daily active dose, depending on the nature and seriousness of the disease and on the characteristics of each patient. Thanks to its dosage flexibility, the drug is employed in stress-dependent, psychiatric and neurologic pathology, in geriatrics and internal medicine, in cases of:

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states of neuropsychic overstrain, maladaptive reactions to stress, acute and chronic psychosocial stress; states of exhaustion and biological stress determined by consumptive and hypercatabolic diseases, convalescence etc.; neurotic disorders: attention and memory disturbances, concentration troubles, the decline of intellectual efficiency; lower effort capacity, physical and psychic asthenia, overwork, exhaustion, sleep disorders; neurovegetative troubles, emotional lability, depression; stress-related disorders and stress-related illnesses: ischemic heart diseases, moderate arterial hypertension; hepatic insufficiency, chronic pancreatopathy; dyslipidemia; as adjuvant therapy in diabetes mellitus and its compli- cations; psychogenic sexual dysfunctions: impotence, frigidity etc.; dysendocrine states by disturbances of hypothalamo-hypophysial complex, in menopause and andropause; cerebral toxic pathology: acute and chronic alcoholism (alcohol abuse and dependence, withdrawal syndrome, psychic and neurologic impairments due to alcoholism, detoxification cures); abuse and dependence on other psychoactive substances; intoxications with carbon monoxide or organic solvents; chronic cerebral circulatory insufficiency and other forms of cerebral vascular pathology: hypoxic, ischemic, thromboembolic, atherosclerotic; neuropsychic troubles caused by head trauma, post-traumatic stress disorders; involutive syndromes during pre-senescence and senescence: poor resistance to stress, physical and psychic tiredness, attention and memory deficiences, cognitive impairments, emotional lability, depression, diminishing of the adaptive capacity etc.

[0072] The administration of the drug, increases the social adaptive capacity, which has been decreased by stress, stress-related disorders and illnesses, psychiatric and neurologic diseases or in senescence, due to its therapeutic actions of behavioural and emotional-affective stabilization and of supporting the cognitive functions.

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Table 1. Neuro-psycho-biological and etio-pathogenic-therapeutic trivalent experimental pattern for the application and checking-up of the anti-stress, anti-impairment and anti-aging drug, in accordance with this invention.

Stress 1. PSYCHIC stress 2. BIOLOGICAL 3. GLOBAL stress etiologies ( P.S. ) stress (G.S.) ( by chronic painful ( B.S. ) (by chronologically Group emotion ) (by chronic alcohol accumulated wear types intoxication) and tear)

I. -age : -age : -age : .8-9 months; .8-9 months; .8-9 months; - methodologies: - methodologies: - methodologies: NORMAL a. cell isolation a. cell isolation a. cell isolation GROUP b. neuromorphology b. neuromorphology b. neuromorphology c. neurobiochemistry c neurobiochemistry c neurobiochemistry (healthv brains) ~ no- °f animals : -no. of animals : - no. of animals : a. 30 rats a. 30 rats a. 60 rats b. 10 rats b. 30 rats b. 90 rats c. 20 rats c 120 rats c 90 rats

JJ# -age: -age: -age: .8-9 months + • 8 - 9 months + . 24-26 months . P.S.; • B.S.; (G.S.); ESCAPACI- - methodologies: - methodologies: - methodologies: TATED ceU 'so,at'on *• ce" 's°lat'on a. ce" isolation b. b. TROUP b. neuromorphology neuromorphology neuromorphology c. neurobiochemistry ' c- neurobiochemistry c neurobiochemistry - no. of animals : - no. of animals : - no. of animals : (cronically a. 40 rats a. 40 rats a. 80 rats stressed brains) 5 2o rats b. 30 rats b. 90 rats c. 20 rats c. 120 rats c 90 rats III. "age: "a8e: -a«e: .8-9 months + .8-9 months + 24-26 months REVTTALI- . . anti-P.S. tr. + . B.S. + (G.S.) + ZED -p-s-i . anti-B.S. tn; . anti-G.S. tr.; CROUP -methodologies: -methodologies: -methodologies: a. cell isolation a. cell isolation a. cell isolation b. neuromorphology b. neuromorphology b. neuromorphology (anti-stress, c. neurobiochemistry c. neurobiochemistry c. neurobiochemistry anti-impairment - no. of animals : - no. of animals : - no. of animals : and anti-aging a- 4U ™is a. 40 rats a. 80 rats incapacitated b- 20 rats D- 30 rats b. 90 rats 20 brains) c. rats c. 120 rats c 90 rats Partial sum 220 animals 560 animals 760 animals TOTAL 1 5 4 0 animals

- anti-P.S. treatment and P.S.: daily, for 2 weeks; - B.S. and anti-B.S. treatment: daily, for 2 months; - G.S. (wear and tear accumulated gradually over 17 months) no.: number; and anti-G.S. treatment: daily, for 2 months. tr.: treatment.

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Table 2. Results of nucleic acid assays (60 animals / gruop) and protein assays (60 animals / gruop) from the brains (telencephalon and diencephalon) of normal rats (age: 8-9 months), incapacitated by biological stress (chronic alcohol intoxication) and therapeutically revitalized with anti-stress, anti-impairment and anti-aging drug, in accordance with this invention. Brains Therapeutically Effici- (groups) Normal Incapacitated by revitalized ency of (age : 8 - 9 months) biological stress Informational i (recapacited) revita- i macromolecules + ( x ± S.D.) lization . ( X S.D.) ( x ± S.D.)

1. Total 1.361+0.098 1.276±0.107 1J31±0.100 DNA incapacitation recapacitation (mg/g (-) 0.085 (+) 0.055 mg wet sb.) mg (-) 63 % (+) 4.1 % significance p<0.001 p<0.01 64.7% 2. Total 2.609±0.117 1.884±0.114 RNA 2.502±0.122 (mg/g incapacitation recapacitation wet sb.) (-) 0.725 mg (+) 0.618 mg (-)27.8 % (+)23.7 % significance p<0.001 p<0.001 85.2 % 3.TP 98.61+2.22 83.40+2.23 95.45±239 (mg/g wet sb.) incapacitation recapacitation 12.05 (-)15.21 mg (+) mg (-)15.4 % (+)12.2 % ; significance p<0.001 p<0.001 79.2 % 4. WIP 23.92+0.72 30.15±1.51 25.84±0.81 (mg/g wet sb.) incapacitation recapacitation (+) 6.23 mg (-) 431 mg (+)20.7 % (-)14.3 % significance p<0.001 p<0.001 69.2 % 5. WSP 74.58±1.85 53.09±1.59 69.56±1.41 (mg/g wet sb.) incapacitation recapacitation (-)21.49 mg (+) 16.47 mg (-)28.8 % (+)22.1 % significance p<0.001 p<0.001 76.6 %

wet sb.: wet substance; - for indicators nos. 1, 2, 3 and 5 TP : total proteins; normal brains = 100%; WIP : water insoluble proteins; - for indicator no. 4 WSP : water soluble proteins; incapacitated brains = 100%.

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Table 3. Results of the calculated indicators derived from the ratios between informational macromolecules, extracted from the brains (telencephalon and diencephalon) of normal rats (age: 8-9 months), incapacitated by biological stress (chronic alcohol intoxication) and therapeutically revitalized with anti-stress, anti-impairment and anti-aging drug, in accordance with this invention.

Brains (groups) Therapeutically Effici- Normal Incapacitated by revitalized ency of (age : 8 - 9 months) biological stress Informational (recapacited) revita- macromolecules . (x + S.D. ) ( i ± S.D.) lization ratios j ( x ± S.D.)

1. 1.919±0.095 1.485+0.072 1.880±0.084 RNA/DNA (ug/ug) incapacitation recapacitation (-) 0.434 (+) 0.395 (-)22.6 % (+)20.6 % significance p<0.001 p<0.001 91.0 % 2. I) WIT 13.787±0.771 15.227+0.949 13.922±0.805 (ug/mg) incapacitation recapacitation (+) 1.440 (-) 1.305 (+) 9.5 % (-) 8.6 % significance p<0.001 p<0.001 90.6 % 3. RNA/TP 26.413+0.945 22.568±0.998 26.197±0.822 (ug/mg) incapacitation recapacitation (-) 3.845 (+) 3.629 (-)14.6 % (+)13.7 % significance p<0.00l p<0.001 94.4 %

(WIP/TP) 24 .3±0.7 % 36.2±1.8 % 27.110.9 % x 100 (%) incapacitation recapacitation (+)11.89 (-) 9.08 (+)32.9 % (-)25.1 % significance p<0.001 p<0.001 76.4 %

TP : total proteins; - for indicators nos. 1 and 3 WIP: water insoluble proteins; normal brains = 100%; WSP: water soluble proteins; for indicators nos. 2 and 4 incapacitated brains = 100%.

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Table 4. Results of the nucleic acid assays (60 animals/group) and protein assays (30 animals/group) from the brains (telencephalon and diencephalon) of normal rats (age: 8-9 months), incapacitated by global stress (wear and tear accumulated gradually over 17 months) and therapeutically revitalized with anti-stress, anti-impairment and anti-aging drug, in accordance with this invention.

Therapeutically Effici- Normal Incapacitated by revitalized ency of (age : 8 - 9 months) global stress revita- Informational (recapacited) lization (macromolecules ( x + S.D.) ( x ± S.D.) ( x ± S.D.)

1353+0.102 1. Total 1.372+0.121 1.328±0.100 DNA incapacitation recapacitation (mg/g (-) 0.044 mg (+) 0.025 mg wet sb.) (-) 3.2 % (+) 1.8 % significance p<0.05 p>0.05 (NS) 56.8 % 2. Total 2.67010.114 2.192±0.123 2.584±0.121 RNA (mg/g incapacitation recapacitation wet sb.) (-) 0.478 mg (+) 0.392 mg (-)17.9 % (+)14.7 % significance p<0.001 p<0.001 82.0 % 3.TP 100.22±2.14 88.48±2.01 96.20±2.02 (mg/g wet sb.) incapacitation recapacitation (-)11.74 mg (+) 7.72 mg (-)11.7 % (+) 7.7 % significance p<0.001 p<0.001 65.8 % 4. WIP 24.72+1.13 30.41±1.12 27.8510.76 (mg/g wet sb.) incapacitation recapacitation (+) 5.69 mg (-) 2.56 mg (+)18.7 % (-) 8.4 % significance p<0.001 p<0.001 45.0% 5. WSP 75.39±139 57.85+1.83 68.48+1.18 (mg/g wet sb.) incapacitation recapacitation (-)17.54mg (+) 10.63 mg (-)23.3 % (+)14.1 % significance p<0.001 p<0.001 60.6 %

wet sb.: wet substance; for indicators nos. 1, 2, 3, and 5 TP : total proteins; normal brains = 100%; WIP : water insoluble proteins; for indicator no. 4 WSP : water soluble proteins; incapacitated brains = 100%.

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lable 5. Results of the calculated indicators derived from the ratios between informational macromolecules, extracted from the brains (telencephalon and diencephalon) of normal rats (age: 8-9 months), incapacitated by global stress (wear and tear accumulated gradually over 17 months) and therapeutically revitalized with anti-stress, anti-impairment and anti-aging drug, in accordance with this invention.

Drains Therapeutically (groups) Normal Incapacitated Effici- revitalized (age: 8-9 months) by global stress ency of 1 iiiiurmauunui i (recapacited) revita- i macromolecules (x ± S.D.) (x 1 S.D.) ratios ', (x ± S.D.) lization

1.944 ±0.078 1.656±0.041 1.917±0.065 RNA/DNA

incapacitation recapacitation (-) 0.288 (+) 0.261 (-)14.8 % (+)13.4 % significance p <0.001 p<0.001 90.6 % I. 13.723±0.771 14.965±0.799 14.022±0.788 (ug/mg) incapacitation recapacitation (+) 1.242 (-) 0.943 (+) 8.3 % (-) 63 % significance p<0.001 p<0.001 75.9 %

26.63210.652 24.76910.913 26.84310.766 (Mg/mg) incapacitation recapacitation (-) 1.863 (+) 2.074 (-) 7.0 % (+) 7.7 % significance p<0.001 p<0.001 111.3 % 4. (WIP/TP) 24.711.1 % 34.411.3 % 28.910.8 % x 100 (%) incapacitation recapacitation (+) 9.70 (-) 5.42 (+)28.2 % (-)15.8 % significance p<0.001 p<0.001 55.9%

I r : total proteins; - for indicators nos. 1 and 3 WIP : water insoluble proteins; normal brains = 100%; WSP: water soluble proteins; ■ for indicators nos. 2 and 4 incapacitated brains =100 %.

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iaoie o. Kesults ot the stages and mechanisms (I-IX) of brain therapeutic revitalization (cellular, subcellular and macromolecular) achieved by the anti-oxidative and anti-catabolic stress actions (1-6) of the drug, in accordance with this invention.

Action Stage and mechanism of therapeutic revitalization i. anti-cataboiic pluriorganelle protection(mitochondriai mainly), Anti- by inhibition of the lipofuscin pigment formation (wear and tear pigment, tertiary lysosomes, old lysosomes, nonfunctional insoluble peroxidated action subcellular wastes).

2. Ill - breaking up of the aggregated pigment conglomerates, 'Neuronal ~ by successive disintegration. linofuscinolytic rrri 111 - vacuolation■ of the nonfunctional, insoluble subcellular residues, act,on - by solubilization.

3^ IV - accumulation in the neurosomal peripherical zones of the subcellular wastes previously processed (broken and Neuron up vacuolated), - by intracellular transport into the neuron. glial lipofuscin ~ — ; " transfer e^mmationoftneP^essedsuDCe,mlarres'^ - by neurono-glial intercellular transfer. VI - collection of the transferred subcellular wastes into the perineuronal glias (particularly into microglias, then into astrocytes and oligodendrocytes) and their intraglial processing (vacuolation), - by glial intracellular transport and by solubilization, respectively.

4. VII - removal of the vacuolated subcellular residues from the perineuronal Glial glias, their gathering in the pericapillary glias and areas (in the vascular lipofuscinolytic end-feet of astro- and microglias and in the pericytes) and continuation of the action intraglial processing (vacuolation), - by glial intracellular transport, by glio-glial intercellular transfer or by- migration of the microglias towards the vascular pole of the neuropil, and by solubilization, respectively. 3. Vni - elimination of the vacuolated subcellular wastes from pericapillary Capillary glias and areas, and their accumulation in the endothelial cells, lipofuscinolytic - by glio-endothelial intercellular transfer. action 5. LX - processing of the transferred subcellular residues (vacuolation) and their Capillary removal from the endothelial cells, lipofuscin - by solubilization and by their outflow into the capillary lumen, elimination respectively.

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Iable 7. Results of the stages and mechanisms ( I - VIII ) of brain therapeutic revitalization ( cellular, subcellular and macromolecular ) achieved by the actions ( 1 - 5 ) with anabolic anti-stress consequences of the drug, in accordance with this invention.

Action Stage and mechanism of therapeutic revitalization dispersed anabolic monoorganelle repopulation with tree nbosomes and 1. rough endoplasmic reticulum, by regulation and activation of the nerv e cell genetic apparatus. Action of - [J - intensive anabolic monoorganelle repopulation, by the emergence ot primary anabolic some subcellular areas of massive regeneration with free ribosomes and organelle rough endoplasmic reticulum, accompanied by hyperfunctional regeneration in diversification of the reticulum (parallel disposing, anastomosing, dilatation), neurosome hyperactive - by enhancement of ribonucleic acid biosynthesis and protein biosynthesis.

Action of JJJ - intensive anabolic polyorganelle repopulation, concentrated upon three secondary groups of subcellular organelles: anabolic (free ribosomes, rough anabolic endoplasmic reticulum, Golgi apparatus), energetic (mitochondria) and contractile (neurofilaments, microtubules), organelle - bv stimulation of the functional and structural anabolism. regeneration in neurosome 3. of anabolic (increase of their functional and life Action of |Y - protection organelles periods), anabolic - by achievement of the optimal repartition and ratios between the organelle anabolic process organelles, and also between these and the catabolic, protection in contractile and energetic organelles. neurosome revitalization of the and of the afterent 4. axoplasm organelles, by normalization of axoplasmic flow and as a consequence of the Action of neurosomal subcellular organelle regeneration (stages I-IV). axonal yjVI - normalization of the myelin sheath, normalization bv regeneration of the oligodendrocyte anabolic subcellular components, which causes the normal and accelerated glial lipoprotein synthesis. 5. VII - normalization of the subcellular organelles from the synaptic knobs, - through axoplasmic flow. Action of VIII - regeneration of the dendritic spines, synaptic - in close correlation w ith the normalization of the terminal buttons (stage normalization VII) and with the revitalization of the neurosomal subcellular components (stages I-IV).

Claims

1. Anti-stress, anti-impairment and anti-aging drug for prophylactic, therapeutic and restorative use, characterized ii

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that it is a synergistic biological neurometabolic and cell-trophic composition, a pro-drug - drug, consisting of

a) 20.1% - 52.3% principles active against oxidative and catabolic stress, namely anti-lipofuscinogenesis, lipofuscinolytic and for lipofuscin elimination, selected from the group consisting of methionine with aminoethanol 5 phenoxyacetates and/or aminoethyl phenoxyacetamides;

b) 7.0%-35.1% principles active against anabolic stress, namely for functional and ultrastructural anabolic regeneration, selected from the group consisting of hydrooxopyrimidine carboxylates and/or oxopyrrolidine acetamides with potassium, zinc and lithium; 10 c) 6.7%-8.6% vasodilative, meaning against hypoxic-ischemic stress, and normolipidemic active principles, consisting of nicotinic alcohol and/or acid, or its derivatives, with magnesium and iodine;

d) and e) 16.6%-19.9% energo-active and anti-toxic active principles, selected from the group consisting of 15 aspartate, fructose, vitamin B1, vitamin B6, monoacid phosphate and sulfate.

2. Anti-stress, anti-impairment and anti-aging drug, according to claim 1 , characterized in that it comprises the following active principles: 18.0%-19.1% methionine; 31.0%-33.1% meclofenoxate or other aminoethanol phenoxyacetates, like meclosulfonate, eclofenoxate, adafenoxate in a weight ratio of 1/0.65-1/0.72, 1/0.79-1/0.83 and 1/0.20- 20 1/0.26 respectively, compared to the amount of meclofenoxate, and/or aminoethyl phenoxyacetamides, like mefexamide, fenoxedil, fexicaine, fipexide in a weight ratio of 1/0.2.4-1/0.29, 1/0.07-1/0.08, 1/0.10-1/0.13 and 1/0.24- 1/0.29 respectively, compared to the amount of meclofenoxate, 18.8%-20.0% hydrooxopyrimidine carboxylates like orotic acid or within the same limits orotates of bioelements of the drug and/or 28.3%-31.4% piracetam or other oxopyrrolidine acetamides, like oxiracetam, etiracetam, dimethylphenyl piracetam, pramiracetam or aniracetam in 25 a weight ratio of 1/0.39-1/0.44, 1/0.28-1/0.34, 1/0.15-1/0.16, 1/0.44-1/0.48 and 1/0.50-1/0.53 respectively, compared to the amount of piracetam; 2.5%-2.9% potassium; 0.31%-0.7% zinc; 0.025%-0.033% lithium; 5.2%-6.6% nicotinic alcohol and/or acid, or its derivatives, like nicotinamide or magnesium nicotinate; 1.5%-1.9% magnesium; 0.005%-0.007% iodine; 9.7%-1 0.6% aspartate; 1 .7%-2.0% fructose, 0.90%-1. 3% Vitamin B1; 1.21%-1 .50% Vita- min B6, 2.6%-3.5% monoacid phosphate, 0.5%-1.0% sulfate; the percentages being related to 100 g of minimum 30 daily active dose.

3. Anti-stress, anti -impairment and anti-aging drug according to claim 1 , characterized in that its active principles are preferably comprised in two pharmaceutical units, complementary regarding the bioavailability and the therapeutic action of the drug, namely: 35 a) a gastrosoluble capsule or coated tablet comprising: 240 mg-280 mg meclofenoxate hydrochloride, 135 mg - 165 mg DL methionine, 80 mg - 100 mg anhydrous DL magnesium aspartate, 9 mg - 1 1 mg D(-) fructose, 7 mg - 9 mg Vitamin B1 hydrochloride, 10 mg-12 mg Vitamin B6 hydrochloride, 9 mg - 1 1 mg non-retarded nicotinic acid, with immediate-release and 9 mg - 1 1 mg anhydrous zinc sulfate, and 40 b) an enterosoluble capsule or coated tablet comprising: 290 mg-340 mg anhydrous orotic acid, 9 mg-1 1 mg D(-) fructose, 70 mg - 90 mg retarded nicotinic acid, with prolonged-release, 88 mg - 108 mg anhydrous dipotassium phosphate, 18 mg - 22 mg magnesium oxide, 2 mg-3 mg lithium carbonate and 0. 1 mg-0. 14 mg potassium iodide. 45 4. Process for the manufacture of the drug according to claims 1 - 3, characterized in that, in order to ensure pharmaceutical stability, maximum bioavailability and therapeutic efficiency, it consists in the granulation or compression of the meclofenoxate or of its related compounds in the gastrosoluble unit, and of the retarded nicotinic acid or of its retarded derivatives in the enterosoluble unit, and in their mixing with the other components in powder form, then so their encasing in hard or soft gelatin capsules, the enteric coating of the units containing the retarded vasodilative substance, or their compression followed by coating with a film obtained from a gastrosoluble or an enterosoluble polymer respectively, in aqueous dispersion or in organic solvent solution.

5. Process according to claim 4, characterized in that the retardation of the vasodilative compound in the enterosolu- 55 ble unit, like nicotinic alcohol and/or acid, or its derivatives, is achieved depending on the time necessary for releasing the active substance, either in the form of a retarded granule, using ethyl cellulose as a retardant and diethyl phthalate as a plasticizer in a weight ratio of 1/0. 18-1/0.24, followed by granule coating with Carnauba wax as the second retardant, in a weight ratio of 1/0.47-1/0.53 compared to ethyl cellulose, or in the form of a retarded tablet,

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obtained by compressing the retarded granule.

6. Process according to claim 4, characterized in that the enteric coating is performed on hard or soft gelatin capsules, filled with the enterosoluble unit composition, using the aqueous dispersion of a gastroresistant-enterosoluble polymer, like cellulose acetate phthalate, polyvinyl acetate phthalate, hydroxypropyl methylcellulose phthalate or a methacrylic copolymer, in a mixture with a plasticizer selected from the group consisting of diethyl phthalate, dib- utylphthalate, propyleneglycol or triacetin , the weight ratio enterosoluble polymer to plasticizer being within the range of 1/0.33 - 1/0.39 and the weight ratio enterosoluble polymer to colorant being within the range of 1/0.0010- 1/0.0012, as coating procedure using a fluidized bed, at an average temperature of 70°C, alternating the spraying and drying stages, and thus obtaining a final coating film of about 60 mg-70 mg/capsule.

7. Use of the drug according to claims 1 - 3 for the manufacture of a medicament for prophylactic medicine for the support and increase of the adaptive capacity and neuropsychic and biological resistance, for simultaneous anti-stress protection of nervous and endocrine systems, liver, heart and blood, and for correction of the main ultrastructural and metabolic imbalances during stress-impairment-aging:

a) by the increase in overall vitality of the body, in the capabilities for physical and psychic efforts, and in the resistance against overstrain, stress and accelerated wear and tear, and

b) by the rapid recovery from the fatigue states, after prolonged efforts or sleeplessness

8. Use of the drug according to claims 1 - 3 for the manufacture of a medicament for therapeutics and restorative medicine for stress-dependent pathology and in accelerated aging, more particularly in:

a) neuropsychic overstrain states, maladaptive reactions to stress, acute and chronic stress, stress-related accelerated wear and tear, and

b) attention and memory disturbances, concentration troubles, decline of intellectual efficiency, lower effort capacities, neurovegetative troubles, depression;

c) ischemic heart diseases, moderate arterial hypertension, hepatic insufficiency, chronic pancreatopathy, dyslipidemia;

d) impotence, frigidity, andropause, menopause;

e) acute and chronic alcoholism, abuse, withdrawal and dependence of psychoactive substances;

f) chronic cerebral circulatory insufficiency and other forms of cerebral vascular pathology, hypoxic, ischemic, thromboembolic, atherosclerotic;

g) involutive syndromes during normal and pathological pre-senescense and senescence.

Patentanspruche

1. AntistreB-, Anti-Beeintrachtigungs- und Antialterungsmittel zum prophylaktischen, therapeutischen und restaurati- ven Einsatz, dadurch gekennzeichnet, da 6 es eine synergistische, biologische, neurometabolische und zelltrophe Zusammensetzung, Prodrug-Arzneimittel ist, bestehend aus:

a) 20,1% - 52,3% gegen oxidativen und katabolen StreB wirksamer Prinzipien, namlich Anti-lipofuscinogene- tische, lipofuscinolytische und zur Elimination von Lipofuscin geeignete, ausgewahlt aus der Gruppe bestehend aus Methionin mit Aminoethanol-Phenoxyacetaten und /oder Aminoethylphenoxiacetamiden;

b) 7,0% - 35,1% aktive Prinzipien gegen anabolen StreB namlich fur die funktionelle und ultrastrukturelle ana- bole Regeneration, ausgewahlt aus der Gruppe bestehend aus Hydrooxopyrimidin-Carboxylaten und /oder Oxopyrrolidinacetamiden mit Kalium, Zink und Lithium;

c) 6,7% - 8.6% Vasodilativa, wirksam gegen hypoxisch-ischamischen Stress, und normolipidamisch aktive Prinzipien, bestehend aus Nicotinsaurealkohol und /oder - Saure oder seinen Derivaten, mit Magnesium und

34 EP 0 804 239 B1

Jod;

d) und e) 16,6% - 19,9% energo-aktiv und antitoxisch wirksame Prinzipien, ausgewahlt aus der Gruppe bestehend aus Aspartat, Fructose, Vitamin B1 , Vitamin B6, Monophosphorsaure und Sulfat; 5 2. Anti-StreB, Anti-Beeintrachtigungs- und Anti-Alterungsmittel nach Anspruch 1 , dadurch gekennzeichnet, daB es die nachfolgenden aktiven Prinzipien aufweist: 18,0% - 19,1% Methionin; 31 ,0% 33,1% Meclofenoxat oder andere Aminoethanolphenoxyacetate, wie Meclosulfonat, Eclofenoxat, Adafenoxat in einem Gewichtsverhaltnis von 1/0,65-1/0,72 1/0,79-1/0,83 und 1/0,20-1/0,26 zur Menge Meclofenoxat, und /oder Aminoethylphenoxyacetamide, 10 wie Mefexamid, Fenoxedil, Fexicain, Fipexid in einem Gewichtsverhaltnis von 1/0,24-1/0,29, 1/0,07-1/0,08, 1/0,10- 1/0,13 und 1/0,24-1/0,29 zur Menge Meclofenoxat; 18,8%-20,0% Hydrooxipyrimidin-Carboxylate wie Orotsaure oder innerhalb der gleichen Grenzen Orotate von Bioelementen des Mittels, und/oder 28,3%-31 ,4% Piracetam oder andere Oxopyrrolidin-Acetamide, wie Oxiracetam, Etiracetam, Dimethylphenyl Piracetam, Pramiracetam oder Aniracetam in einem Gewichtsverhaltnis von 1/0,39 - 1/0,44, 1/0,28 - 1/0,34, 1/0,15 - 1/0,16, 1/0,44 - 1/0,48 und 15 1/0,50 - 1/0,53 zur Menge Piracetam; 2,5% - 2,9% Kalium; 0,3% - 0,7% Zink; 0,025% - 0,033% Lithium; 5,2%-6,6% Nicotinalkohol und/oder -saure oder seine Derivate, wie Nicotinamid oder Magnesium-Nicotinat; 1,5% - 1,9% Magnesium; 0,005% - 0,007% Jod; 9,7% - 1 0,6% Aspartat; 1,7% - 2,0% Fructose; 0,9% - 1 ,3% Vitamin B1 ; 1 ,2% - 1 ,5% Vitamin B6; 2,6% - 3,5% einbasisches Phosphat ; 0,5% - 1 ,0% Sulfat; wobei sich die Prozentsatze auf 100g der minimalen taglichen aktiven Dosis beziehen. 20 3. AntistreB-, Anti-Beeintrachtigungs- und Anti-Alterungsmittel nach Anspruch 1 , dadurch gekennzeichnet, daB seine aktiven Prinzipien bevorzugt in zwei pharmazeutischen Einheiten enthalten sind, die hinsichtlich der Bioverfiigbar- keit und der therapeutischen Wirkung des Mittels komplementar sind, namlich:

25 a) eine magensaftlosliche Kapsel oder iiberzogene Tablette, aufweisend: 240 mg - 280 mg Meclofenoxat- Hydrochlorid, 135 mg - 165 mg DL Methionin, 80 mg - 100 mg wasserfreies DL-Magnesiumaspartat, 9 mg - 1 1 mg D(-) Fructose, 7 mg - 9 mg Vitamin B1 Hydrochlorid, 10 mg - 12 mg Vitamin B6 Hydrochlorid, 9 mg - 1 1 mg Nicotinsaure ohne verzogerte Freisetzung, mit sofortiger Freisetzung und 9 mg - 1 1 mg wasserfreies Zinksulfat und 30 b) eine darmsaftlosliche Kapsel oder iiberzogene Tablette, aufweisend: 290 mg-340 mg wasserfreie Orot- saure, 9 mg-1 1 mg D(-)Fructose, 70 mg-90 mg Nicotinsaure mit verzogerter Freisetzung, 88 mg-108 mg wasserfreies Dikaliumphosphat, 18 mg - 22 mg Magnesiumoxid, 2 mg - 3 mg Lithiumcarbonat und 0,1 mg - 0,14 mg Kaliumjodid. 35 4. Verfahren zur Herstellung des Mittels nach Anspruch 1 bis 3, dadurch gekennzeichnet, daB zur Sicherstellung der pharmazeutischen Stabilitat, maximalen Bioverfiigbarkeit und therapeutischen Wirksamkeit ist aus den Granulaten oder PreBlingen des Meclofenoxats oder seiner entsprechenden Verbindungen in der magensaftloslichen Einheit und der verzogerten Nicotinsaure oder seiner mit Verzogerung freigesetzten Derivative in der darmsaftloslichen 40 Einheit und in deren Mischung mit den anderen Komponenten in Pulverform, anschlieBendes Einkapseln dersel- ben in Hart- oder Weichgelatinekapseln, wobei der darmsaftlosliche Uberzug der Einheiten die vasodilative Sub- stanz mit retardierter Freisetzung enthalt oder deren PreBlinge, gefolgt durch Uberzug mit einem Film erhaltlich aus einem magensaftloslichen oder einem darmsaftloslichen Polymer in waBriger Dispersion oder in Losung in organischem Losungsmittel enthalt. 45 5. Verfahren nach Anspruch 4, dadurch gekennzeichnet, daB die Verzogerung der vasodilativen Verbindung in der darmsaftloslichen Einheit, wie Nicotinalkohol und/oder saure oder seine Derivate abhangig vom zur Freisetzung des Wirkstoffes notwendigen Zeitraum erzielt wird, entweder in Form eines Retard-Granulats, unter Verwendung der Ethylcellulose als Verzogerungsmittel und Diethylphtalat als Weichmacher in einem Gewichtsverhaltnis von so 1/0, 18 - 1/0,24, gefolgt durch Uberzug des Granulats mit Carnaubawachs als zweitem Verzogerer, in einem Gewichtsverhaltnis von 1/0,47-1/0,53 zur Ethylcellulose, oder in Form einer Retard-Tablette, erhalten durch Kom- pression des Retard-Granulats.

6. Verfahren nach Anspruch 4, dadurch gekennzeichnet, daB der darmsaftlosliche Uberzug auf Hart- oder Weichge- 55 latinekapseln aufgebracht, gefiillt mit der darmsaftloslichen Einheitszusammensetzung, unter Verwendung der waBrigen Dispersion eines magensaftresistenten darmsaftloslichen Polymeren, wie Celluloseacetatphthalat, Poly- vinylacetatphtalat, Hydroxypropyl-Methylcellulose-Phthalat oder einem Methacrylsaure-Copolymeren, in einer Mischung mit einem Weichmacher ausgewahlt aus der Gruppe bestehend aus Diethylphthalat, Dibutylphthalat,

35 EP 0 804 239 B1

Propylenglycol oder Triacetin, im Gewichtsverhaltnis von darmsaftloslichem Polymer zum Weichmacher im Bereich von 1/0,33 - 1/0,39 und dem Gewichtsverhaltnis von darmsaftloslichem Polymer zum Farbstoff im Bereich von 1/0,0010 - 1/0,0012, wobei als Uberzugsverfahren einfluidisiertes Bett eingesetzt wird, bei einer Durchschnittstem- peratur von 70°C, wobei die Sprtih- und Trockenschritte abwechselnd durchgefiihrt werden, wodurch ein endgulti- ger Filmiiberzug von etwa 60 mg - 70 mg/Kapsel erzielt wird.

7. Verwendung des Mittels nach Anspruch 1-3 in der prophylaktischen Medizin, zur Unterstiitzung und Verstarkung der adaptiven Fahigkeiten und neuropsychischen und biologischen Widerstandsfahigkeit fur die gleichzeitige Anti- stressschutz der Nerven und endocrinen Systeme, der Leber, des Herzens und des Blutes, und zur Korrektur der Haupt-Strukturellen und metabolischen Ungleichgewichte wahrend des Alters durch StreB beeinfluBt:

a) Durch Steigerung der Gesamtvitalitat des Korpers in dessen Fahigkeiten, psychischen und physischen Anstrengungen zu widerstehen, und in der Widerstandsfahigkeit gegen Uberbeanspruchung, StreB und beschleunigte Abnutzung und Alterung, und

b) fur die schnelle Wiederherstellung aus Ermudungszustanden nach verlangerten Anstrengungen oder Schlaflosigkeit.

8. Verwendung des Mittels nach Anspruch 1 -3 zur Herstellung eines Arzneimittels fur therapeutische und restaurative Medizin, bei streBabhangigen pathologischen Zustanden und beschleunigter Alterung, insbesondere bei

a) neuropsychischen Uberbeanspruchungzustanden, Maladaptiv-Reaktion gegeniiber StreB, akutem und chromischem StreB, streBabhangige UnregelmaBigkeiten und Krankheiten;

b) Aufmerksamkeits- und Gedachtnisstorungen, Konzentrationsprobleme, Abnahme der intellektuellen Effizi- enz, niedrigere Anstrengungsfahigkeit, neurovegetative Probleme, Depressionen;

c) Ischamische Herzkrankheiten, geringfugige arterielle Hypertension, Leberinsuffizienz, chronische Pancrea- topatie, Dyslipidamie;

d) Impotenz, Frigiditat, Andropause, Menopause;

e) akuter und chronischer Alkoholismus, MiBbrauch, Entzug und Abhangigkeit vom phsycho -aktiven Substan- zen;

f) chronische cerebrale Kreislaufinsuffizienz und andere Formen cerebraler vaskularer Pathologie: hypoxische, ischamische, tromboembolische, atherosclerotische;

g) Syndrome des Zusammenbruchs wahrend normaler und pathologischer Praseneszenz und Seneszenz.

Revendications

1 . Medicament anti-stress, anti-affaiblissement et anti-vieillissement pour I'utilisation prophylactique, therapeutique et reconstituante, caracterise en ce qu'il s'agit d'une composition biologique, neurometabolique et cellulotrophique synergique, d'un promedicament-medicament constitue

(a) de 20,1 % a 52,3 % de principes actifs contre le stress oxydant et catabolique, notamment par anti-lipofus- cinogenese, lipofuscinolyse et pour I'elimination de la lipofuscine, choisis dans le groupe constitue de la methionine avec des phenoxyacetates d'aminoethanol et/ou des aminoethyl phenoxyacetamides ; (b) de 7,0 % a 35,1 % de principes actifs contre le stress anabolique, notamment pour la regeneration anabolique fonctionnelle et ultrastructurale, choisis dans le groupe constitue des carboxylates d'hydrooxypyrimidine et/ou des oxopyrrolidine-acetamides avec du potassium, du zinc et du lithium ; (c) 6,7 % a 8,6 % de principes vasodilatateurs, c'est-a-dire contre le stress hypoxique-ischemique et des principes actifs normolipemiques, constitues d'acide et/ou d'alcool nicotinique ou de ses derives, avec du magnesium et de I'iode ; (d) et e) 16,6 % a 19,9 % de principes actifs energo-actifs et anti-toxiques, choisis dans le groupe constitue de I'aspartate, du fructose, de la vitamine B1 , de la vitamine B6, du phosphate monoacide et du sulfate.

36 EP 0 804 239 B1

2. Medicament anti-stress, anti-affaiblissement et anti-vieillissement selon la revendication 1 , caracterise en ce qu'il comprend les principes actifs suivants : 18,0 % a 19,1 % de methionine, 31,0 % a 33,1 % de meclofenoxate ou d'autres phenoxyacetates d'aminoethanol, tels que le meclosulfonate, I'eclofenoxate, I'adafenoxate dans un rapport ponderal de 1/0,65 a 1/0,72, 1/0,79 a 1/0,83 et 1/0,20 a 1/0,26 respectivement, par rapport a la quantite de 5 meclofenoxate et/ou d'aminoethyl phenoxyacetamides, comme le mefexamide, le fenoxedil, la fexicai'ne, le f ipexide dans un rapport ponderal de 1/0,24 a 1/0,29, 1/0,07 a 1/0,08, 1/0,10 a 1/0,13 et 1/0,24 a 1/0,29 respectivement, par rapport a la quantite de meclofenoxate, 18,8 % a 20,0 % de carboxylates d'hydrooxopyrimidine tels que I'acide orotique ou, dans les memes limites, des orotates de bioelements du medicament et/ou 28,3 % a 31 ,4 % de piracetam ou d'autres oxypyrolidine acetamides tels que I'oxyracetam, I'etiracetam, le dimethylphenyl piracetam, le 10 pramiracetam ou I'aniracetam dans un rapport ponderal de 1/0,39 a 1/0,44, 1/0,28 a 1/0,34, 1/0,15 a 1/0,16, 1/0,44 a 1/0,48 et 1/0,50 a 1/0,53 respectivement, par rapport a la quantite de piracetam ; 2,5 % a 2,9 % de potassium ; 0,31 % a 0,7 % de zinc ; 0,025 % a 0,033 % de lithium ; 5,2 % a 6,6 % d'acide et/ou d'alcool nicotinique ou de leurs derives tels que le nicotinamide ou le nicotinate de magnesium ; 1 ,5 % a 1 ,9 % de magnesium ; 0,005 % a 0,007 % d'iode ; 9,7 % a 10,6 % d'aspartate ; 1 ,7 % a 2,0 % de fructose, 0,90 % a 1 ,3 % de vitamine B1 ; 1 ,21 % a 1 ,50 15 % de vitamine B6, 2,6 % a 3,5 % de phosphate monoacide, 0,5 % a 1 ,0 % de sulfate ; les pourcentages etant rap- portes a 100 g de dose active quotidienne minimale.

3. Medicament anti-stress, anti-affaiblissement et anti-vieillissement selon la revendication 1 , caracterise en ce que ses principes actifs sont de preference compris dans doux unites pharmaceutiques, complementaires en ce qui 20 concerne la biodisponibilite et Taction therapeutique du medicament, a savoir :

a) une capsule gastrosoluble ou un comprime enrobe comprenant : 240 mg a 280 mg de chlorhydrate de meclofenoxate, 135 mg a 165 mg de DL methionine, 80 mg a 100 mg d'aspartate de magnesium DL anhydre, 9 mg a 1 1 mg de D(-)fructose, 7 mg a 9 mg de chlorhydrate de la vitamine B1 , 10 mg a 12 mg de chlorhydrate 25 de la vitamine B6, 9 mg a 1 1 mg d'acide nicotinique non retarde, avec liberation immediate et 9 mg a 1 1 mg de sulfate de zinc anhydre, et b) une capsule enterosoluble ou un comprime enrobe comprenant : 290 mg a 340 mg d'acide orotique anhydre, 9 mg a 1 1 mg de D(-)fructose, 70 mg a 90 mg d'acide nicotinique retarde a liberation prolongee, 88 mg a 1 08 mg de phosphate dipotassique anhydre, 1 8 mg a 22 mg d'oxyde de magnesium, 2 mg a 3 mg de carbo- 30 nate de lithium et 0,1 a 0,14 mg d'iodure de potassium.

4. Procede de fabrication du medicament selon les revendications 1 a 3, caracterise en ce que, pour assurer la stability pharmaceutique, une biodisponibilite maximale et une efficacite therapeutique maximale, il consiste dans la granulation ou la compression du meclofenoxate ou des composes apparentes dans I'unite gastrosoluble, et de 35 I'acide nicotinique retarde ou de ses derives retardes dans I'unite enterosoluble, et dans leurs melanges avec les autres constituants sous forme de poudre, puis leur introduction dans des capsules de gelatine dure ou molle, le revetement enterique des unites contenant la substance vasodilatatrice retardee ou leur compression suivie d'un enrobage d'un film obtenu a partir d'un polymere gastrosoluble ou d'un polymere enterosoluble respectivement, en dispersion aqueuse ou en solution dans un solvant organique. 40 5. Procede selon la revendication 4, caracterise en ce que le retardement du compose vasodilatateur dans I'unite enterosoluble, comme I'alcool et/ou I'acide nicotinique, ou leurs derives, est realise en fonction du temps neces- saire pour liberer la substance active, soit sous la forme d'un granule retarde, en utilisant de I'ethylcellulose comme retardateur et du phtalate de diethyle comme plastifiant dans un rapport ponderal de 1/0,18 a 1/0,24, suivi de 45 I'enrobage du granule avec de la cire de Carnauba comme second retardateur, dans un rapport ponderal de 1/0,47 a 1/0,53 par comparaison avec I'ethylcellulose ou sous la forme d'un comprime retarde, obtenu en comprimant le granule retarde.

6. Procede selon la revendication 4, caracterise en ce que I'enrobage enterique est effectue sur des capsules de so gelatine dure ou molle, remplies de la composition de I'unite enterosoluble, en utilisant la dispersion aqueuse d'un polymere gastroresistant-enterosoluble comme I'acetate phtalate de cellulose, I'acetate phtalate de polyvinyle, le phtalate d'hydroxypropyle methylcellulose ou un copolymere methacrylique, en melange avec un plastifiant choisi dans le groupe constitue du phtalate de diethyle, du phtalate de dibutyle, du propyleneglycol ou de la triacetine, le rapport ponderal du polymere enterosoluble au plastifiant etant dans I'intervalle de 1/0,33 a 1/0,39 et le rapport 55 ponderal du polymere enterosoluble au colorant etant dans I'intervalle de 1/0,0010 a 1/0,0012, en utilisant comme technique d'enrobage un lit f luidise, a une temperature moyenne de 70 °C, en faisant alterner les etapes de pulve- risation et de sechage ce qui fournit un film d'enrobage final d'environ 60 mg a 70 mg/capsule.

37 EP 0 804 239 B1

Utilisation du medicament selon les revendications 1 a 3 pour la fabrication d'un medicament pour la medecine prophylactique pour le soutien et I'augmentation de la capacite d'adaptation et de la resistance neuropsychique et biologique, pour la protection anti-stress simultanee des systemes nerveux et endocrinien, du foie, du coeur et du sang, et pour la correction des desequilibres ultrastructuraux et metaboliques principaux au cours du stress-affaiblissement-vieillissement :

a) par I'augmentation de la vitalite globale de I'organisme, des capacites d'effort physique et psychique et de la resistance contre le surmenage, le stress et I'usure acceleree, et b) par la recuperation rapide des etats de fatigue apres les efforts prolonges ou I'insomnie.

Utilisation du medicament selon les revendications 1 a 3 pour la fabrication d'un medicament pour la medecine therapeutique et reconstituante pour la pathologie dependant du stress et dans le vieillissement accelere, plus parti - culierement dans :

a) les etats de surmenage neuropsychique, les reactions inadaptees au stress, au stress aigu et chronique, a I'usure acceleree liee au stress, et b) les perturbations de I'attention et de la memoire, les troubles de la concentration, la diminution de I'efficacite intellectuelle, I'abaissement des capacites d'effort, les troubles neurovegetatifs, la depression ; c) les maladies cardiaques ischemiques, I'hypertension arterielle moderee, I'insuffisance hepatique, la pan- creatopathie chronique, la dyslipemie ; d) I'impuissance, la frigidite, I'andropause, la menopause ; e) I'alcoolisme aigu et chronique, I'abus, la privation et la dependance de substances psycho-actives ; f) une insuffisance circulatoire cerebrale chronique et d'autres formes de pathologies vasculaire cerebrale, hypoxique, ischemique, thrombo-embolique, athero-sclerotique ; g) des syndromes involutifs au cours de la presenescence et de la senescence normales et pathologiques.

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Fig. 15.

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