SE416266B - SET TO MAKE A PREPARED CONTENTS A PREVIOUS NON-DESCRIPTION HORMON, LITORALON, PRODUCED BY THE BISCOLD CARDS - Google Patents

Description

p 721111027-4 2 This hormone has polypeptide-like structure. It is prepared by extraction with hot hydrochloric acid or phenol from defatted, finely divided parathyroid glands.

From the extract, the crude product is obtained, which can be precipitated 51 at pH ä¿8, by repeated fractionation with one solvent and by salting and precipitation with trichloroacetic acid. This purity is suitable from a therapeutic point of view. The present invention is based on the discovery that the parathyroid glands, in addition to the already known hormones, contain "if one or more substances with hormonal action" are present in the chemically pure calcium active parathyroid hormone and its active isohormones can be produced by a complicated gel filtration technique. , which substances fl, ¿differ significantly from those previously known. The chemical structure of the Litoralon has later been clarified. get net ha; namely Show: to be Z-L-glueamyitaurine. It is thus a question of a micropeptide having molecular weight 254,27 and gross formula: ll¿; fC? Hl4NèO6S. ¿: '_ The new substances with hormonal action are potassium inactive, 2Ö ~ g¿d v s ï contrary to the previously known hormones they affect the uääâ blood calcium level neither positively nor negatively, but raise the blood _; kiseinale. The highly purified fractions already develop in very small g _., f concentrations (below l / ug / kg body weight) pharmacological and therapeutic action over a wide range, which will be described in more detail below. From a chemical point of view, the new substance or the new substances are distinguished by the following characteristic properties: they are not precipitated with trichloroacetic acid, ie they are not unambiguous in some egg whites and do not have a polypeptide nature. They are not in fat-like substances. are not soluble in benzene or chloroform but are, in contrast, water-soluble. Litoralon retains its biologic activity even after a solution thereof is subjected to a treatment which denaturs egg whites, after a treatment with nitric acid or ethylene oxide. .

According to the present invention, the substance litoralone suitable for pharmaceutical and cosmetic use is prepared. It is water soluble, contains, C, H, N, O and S, is neither egg white cells steroid, has a molecular weight of 254.27, has a non-fat character, Flare insoluble in benzene; chloroform and carbon tetrachloride, contains fifl 55 40k 7210027-4 .the following characteristic molecules or molecule fragments, namely C7H14N5O2S and C5H9N5O, have the following physiological effects, namely cause phosphaturia, increase in rats herder glands, also have radiotherapy - element concentration; in clinical-therapeutic use, they favorably affect Sjögren's syndrome, Rhino-laringo-pharingitis sicca, Scleroderma, Skin candidiasis, radiation erythema and other diseases.

In the method according to the invention, a concentrate or a mixture containing the previously not described litoralone hormone is prepared, proceeding in the manner specified in the following claim.

Materials and cell cultures derived from mammalian parathyroid glands are also suitable as starting materials for the production of lithoralone. The concentrate (aqueous solution or lyophilized powder) prepared in this way is already suitable for the preparation of pharmaceutical preparations, with which the pharmaceutical or biological effects described below can be achieved.

More active concentrates can also be prepared by striving for a product which contains the active substance in concentrated or pure form and / or by activating the accompanying calcium-active hormone. The purer products are better suited for the preparation of pharmaceutical preparations, both ethically and from the standpoint of unambiguous dosage. The aqueous extracts can be freed from egg white bodies and polypeptides by adding the extract with egg white precipitants and separating the precipitate.

For the precipitation of egg whites, trichloroacetic acid, sulphosalicylic acid, etc. can be used. Egg whites can also be separated by means of solvents and by salting out. Precipitation with trichloroacetic acid has proven to be suitable and successful. A disadvantage, however, is that it is relatively difficult to remove excess precipitant. This can conveniently be achieved by ether extraction carried out in series, but “one can also heat the solution liberated from egg whites and can in this way destroy 90% of the trichloroacetic acid, for according to recent experience litoralone is not particularly heat sensitive. During this treatment, kioroform and köiaitxia are formed.

According to the experience gained in the present invention, the division of the egg whites, polypeptides and other fractions can be accomplished with good results by gel filtration. In this case, the desired substance can be extracted in relatively high purity in the fraction in question. A very suitable method is purification by gel filtration, after removing the egg whites from the aqueous fraction, whereby a litoralone fraction with a relatively high degree of purity can already be recovered in a column with a high fractionation capacity, possibly by repeated gel filtration of a narrow fraction.

The concentrate can also be purified in many other ways, for example by chromatography, dialysis, extraction, precipitation, adsorption, etc. Combinations of these methods can of course also be used.

For the elimination of the calcium-active parathyroid hormone, it is convenient to carry out treatment with substances which react easily with egg whites and polypeptides. Nitric acid, ethylene oxide, formalin, etc. can be used for this purpose.

As pointed out above, the uniformity of the litoralone has not yet been clarified. It has been established that the material can be divided into different active "litorelone fractions" by various chemical and physicochemical procedures. Nan can thus prepare different fractions by precipitation with solvent, chromatography, gel filtration, dialysis, extraction, fractional salting, treatment with ion exchanger. For a finer separation of the fractions, a combination of these methods can be used.

Litoralon or litoralone-containing fractions are mixed with biologically inert carriers in the preparation of therapeutic and / or preventive preparations. The content of active ingredient in these preparations can vary within wide limits and depends, among other things, on the intended use. Thus, the active ingredient may be included in the preparation at, for example, about 0.05-10 mg per dose. These preparations can be prepared in a manner well known in the pharmaceutical industry from litornlon-containing, solid or liquid concentrates or mixtures in the form of tablets, snippets, salaries, ointments, powders or sprays. Tablets with 0.05-3 mg active ingredient can be prepared by wet granulation, but dry procedures can also be applied. As carrier and excipient, any common inert substances used for the manufacture of medicinal products can be used. Injectable solutions can be prepared in the form of aqueous solutions containing 0.05-5 mg / ml. They must, of course, be free from egg whites and, moreover, they must be completely free from the additives by which egg whites have been removed. It is convenient to prepare injectable solutions by sterile filtration. The solutions have no irritating effect and do not cause anaphylactic reactions.

Ointments with 0.01-0.1 g / kg active ingredient can also be prepared. For this purpose, most hydrophilic and hydrophobic salt bases can be used. The same applies to the production of suppositories and cocoa butter can be used as well as synthetic suppository bases, for example carbovax. The amount of active ingredient in suppositories is 0.05-5 mg depending on the purity of the concentrate.

Cosmetic preparations containing litoralone can also be prepared. These can be prepared from solid or liquid litoralone concentrates with known solvents and additives, fillers, diluents, lubricants and vehicles in the form of ointments, solutions, creams, emulsions, powder mixtures, sprays, etc. The active ingredient content is suitably 0.01 -0.5 mg / g.

With litoralone-containing preparations, the following very extensive clinical-therapeutic trials have been performed. The preparation was generally administered 5 times orally in doses of 0.4-0.5 mg of litoralone-containing concentrate. Harmful side effects were practically not observed in trials of approximately 5000 people. In a few cases, a urticaria-like rash was observed on the skin surface and in a few cases, nausea and feelings of vomiting were reported. These complaints occurred in a maximum of 5% of the treated patients.

I Characteristic of the hormonal effects of the new substance (s) - are physiological effects: A a) Pharmacological effect: The blood calcium level is practically not affected. The silicon content of the blood is increased and the concentration of other essential trace elements changes, for example Cu, F etc.

Phosphaturia is caused as well as an increase in the level of adenosine monophosphate (AMP) and cyclic adenosine monophosphate (cAMP) in the urine.

Lowering blood sugar levels. lg Impact of factors affecting blood coagulation. Lypolysis increase.

Impact on the hypothalamic-pituitary-adrenal system.

(Induction of an adaptation syndrome-resistance state: Increase in serum cortisol levels, sensitization of the adrenal cortex to ACTH, protection against stressors caused by steroid overproduction in the adrenal cortex, inhibition of pituitary release of endogenous ACTH, protection against adrenal hypertrophy) In long-term (one-year) dosing, mild hypertrophy occurs in the glomerular zone, but the fascicular zone shows weak hypofunction, which has been established by fine histological examinations. The myeloid-erythroid ratio in the bone marrow is increased. _ gene enzyme activating effect: Activation of the growth and fibroblast growth as well as the building up of the connective tissue population and the fibrillal fibrous system, improvement of wound healing prevented by cortisone.

Elevation of glycoprotein and seromucoid heroin in blood serum.

Healing of bone and cartilage tissue and in small concentrations activation of osteoblasts and chondrocytes. The hormone is necessary for the physiological function of the bone foundations as well as cartilage and joint tendons, ligaments and synovial membranes. By intracellular action, an increase in the electron density in Golgi's Apparato and the endoplasmic reticulum in the cells, which build up bone and cartilage tissue. Defense against experimental laziness. Decrease in serum citrate concentration and increase in alkaline phosphatase concentration.

Protection of the organism against the harmful effects of electromagnetic radiation. Reducing the recovery time for hand rhythms and significantly increasing the lifespan of rats irradiated with a lethal dose of X-rays. Reduction of the mitochondrial effect in the bone marrow myeloids caused by X-rays. Not only prophylactic but also therapeutic effect against radiation damage.

Significant increase in the weight of the Harder glands in rats that are extraretinal photoreceptors.

Influence of the biological-energetic effect of light radiation.

Increased uptake of C14-labeled glucosamine into the renal tubules and significant increase in the mucopolysaocaride concentration in the renal tissue. Increase in diuresis.

Significant reduction in the temperature drop in the rectum caused by histamine. Regulatory effect on arteriovenous anastomosis.

Effect on the endothelium in the walls of the veins and on the mucopolysaccharide synthesis in the intimate. b) Clinical effect. 1) In relation to pathological changes exerted effect on the organism's total mucosal and respiratory system: Sicca syndrome in the eyes Beginning emphysema Disorders of the mucous and salivary gland secretions 40 ß IX) \, "l 40) 7 'YÉHDOZT-ë- Leukokeratosis and leukoplakia in the mucous membranes Afte recurrence and Behcet's disease Candidates in the mucous membranes Plumer-Vinson syndrome Syndrome syndromes Reiter syndrome of radiation Other disease states fl v ectodermal origin: Skin candidiasis Verruca juvenílis and vulgaris Kondyloma acuminetum Keratosis palmoplantaris Acanthosis nigricans Scleroderma mor, morphea Sclerodema Other collagenoses Necrobiosis lipoidíka Granuloma annulnre Penis plupuru ecturium osus atrophiomns Radiodermen Alopecia Onychodystrofia Tele gi ngiectasien, Cutis marmorats Psoriasis pustulosn Impetigo herpetiformís Iktyos Disorders of pigmentation, Vitiligo Diseases of the musculoskeletal system and skeletal bones ~ q: De s.k. degenerative pathogens due to wear and tear: Osteoarthritis Dysoscopathy Spondylosis Other pathogens Bechterew's disease Osteogenesis imperfecta Idiopathic Osteoporosis Other Osteoporoses 40 40 vzlnozv fl a Q 4)) 6) 7) Osteofibrosis Osteopathic Neuropathic Neuropathy endocrine origin (vegetative dysbonia, mental symptoms associated with menopause, mental symptoms occurring after strumectomy, Cushing's psychoses, etc.) Depressive decompensation nv neurotic origin or character, state of impulse deficiency, inactivity or tenebröst condition.

Kntntoni Bekgrundsternpi at weaning from abuse and nsrkomnní.

Obstetrics and gynecological diseases: Blood poisoning during pregnancy (Hypertension, edema, albuminuria preeclampsia, eclampsia, accumulative toxicemia).

Henstrual disorders (Metropolitan, amenorrhea, rsromenorre, polymenorre, etc.) Menopausal syndrome (Hypertension, emotions, sweating, mental symptoms, disturbances in the circadian serum cortisol change).

Pruritus or craurosis vulvae Mastopnti fibrocyshicø Erosio colli Laktntionsstörningsr androgenic preponderance hypogonadism Changes connected sirwer f rg al nerz Cataract fi incipiens Sklerotískn changes in Fundus Retinopathy diabetíca and Glnuooma diabetica Disturbances of taste and smell Otosclerosis Otitis recidivans _ vocal cord damage, hoarseness of organic origin Thermo Regulatory disorders: Hype- or hyperhydrosis Hyperhydros palmoplantaris Frossa, echauffering .1 55 46 s) 9)) 41) 12)) 9 7210027-4 Subfebrility of endocrine character Peripheral arteriovenous anastomotic dysfunction Raynaud's disease Acrocyanosis Dental and oral diseases: Periodontitis Hypothalamic tenderness in dental teeth Vascular disorders: Retinopathy diabetic Ulcer cruris Arterosclerosis of the obliter Endocrine disease pictures Juvenile Bnsophilism Cushing's disease and syndrome, nephrosclerosis of endogenous origin, Secondary Cushing's disease Cretinism Fagge's disease Hyp oparathyroidism adiposis dolorosa Lypodystrophia adiposogenítalis Lypodystrophía progressive Cortícogenitalis syndrome late diabetic syndrome Morgagni syndrome Gnstroenterologiske syndromes: Hyp0 ~ or nnncídit f t Large torpid svulstev atrophic gastritis Stràlningsskydd and protection against external environmental impact: Stràlningsskydd therapy of radiation injury Meteoropati spacecraft Aviation iatrogenic damage: Protection against the harmful side effects of radiomimetic and Immuno ~ Suppression substances Treatment of steroid damage with glycocorticoid effect Protection against antibody damage caused by orally administered steroids '15 50 55ï 40 721002? -Å Q Treatment of iatrogenic cataract Phenorosis 44) Gerontological diseases: Progeria Geroderma Symptomatic and guichromic Many syndromes associated with general mesoectodermal injuries or acquired.

(Dermatological, ophthalmological, psychiatric endocrinological, pediatric, organic, pulmonological, gynecological, urogenital, etc.).

It should be noted that the above-mentioned pharmacological and clinical effects are only some characteristic effects, which can be obtained with the new hormone.

The pharmacological and chemical examinations have been carried out systematically for the past three years at 8 university clinics, state institutes and pharmaceutical factories, and the clinical examinations have been carried out systematically at 20 university clinics, state institutes and hospitals in Budapest for three years. back. Therapeutic examinations of various diseases have so far been carried out on a registered patient material consisting of 5,000 people.

The therapeutic effect on some disease states has been determined after several weeks or a few months of treatment (Bechterew's disease). However, there are also relatively rapid effects (eg Ozaena). In some cases the remission is persistent, in some cases strongly hormone dependent and after treatment discontinuation in patients rapid relapse (eg Rhinopharingitis sicca).) The pure fractions exert the therapeutic effect even at very low concentrations. (At a concentration of 0.001 ppm or less, corresponding to an oral dose of 1 microgram / kg body weight.) It is surprising that litoralone has a beneficial therapeutic effect on such a large number of apparently incoherent diseases. In connection with this, reference may be made to cortisone and derivatives thereof, which likewise have a hormonal action and have a therapeutic effect on several hundred diseases. With regard to its great importance, the therapeutic effect of Litoralon can only be compared with the cortisone derivatives and opens a new chapter in therapy, pharmacology and biochemistry. However, it differs from the cortisone derivatives not only because of the significantly different indications (these are in many cases of opposite effect) but also because of its significant '40 50 55 40 in telets, i.e. 1 "' 7210027-4 lower toxicity and In general, litoralone has a beneficial effect that develops gradually, thereby distinguishing litoralone from cortisone, which often has a rapid effect, especially in chronic, degenerative, sclerotic progressions and not during exacerbation during flare-ups, during which During clinical use, even during very long-term treatment (one year) it has not been possible to observe any significant, harmful side effects.

The working hypothesis in the discovery of the various disease pictures, which do not seem to be in any connection with each other, has been the following: The discovery of the "evolutionary therapeutic theory" has in principle proved to be helpful and has been indicative. The essence of the theory is explained as follows: The fact that the parathyroid glands have not yet developed as endocrine organs in the invertebrates and in the fish and during the development occurs in the reptiles at the same time as the bronchial apparatus disappears, ie has its origin associated with the incipient life on land and histologically developed from the pharyngeal epidemic which during life in water serves as a respiratory organ and which during development has become superfluous, has given rise to the assumption that the function of the parathyroid glands is adaptation of the organism originally living in water to life on land.

Life on land began to develop on Earth millions of years ago and thus a number of basic physiological problems had to be solved, which arose during the transition from water to land. The respiratory system changed in principle. With the formation of the lungs, the reduction of the surface tension of the pulmonary alveoli must be solved by the production of surfactants, so that the alveoli do not fall into an atectic state and breathing is facilitated. Thus, it must be ensured that the inhaled air had optimal temperature and humidity and was free from mechanical contaminants. The cornea and conjunctiva of the eyes must be protected against dehydration and against microtraumas caused by stump and air pollution. In addition, the eyes must be adapted to the ratio between the refractive index of the air and the cornea and the eyeball lubricated when rotated. The lacrimal glands and other glands developed.

Furthermore, the problem must be solved how the dry nutrients could be introduced into the stomach (formation of mucin-containing skewers).

The auditory and olfactory organs were adapted to the new biosphere.

The bands received a fundamentally new function. It must protect the body against dehydration, radiation damage and external damage.

Thermal regulation was given its important task. The seawater had a joint 55 ao 7210027-4 1: practically constant temperature in comparison with the extreme temperature changes on land. Sweat and sebaceous glands, pigment cells and additional ectodermal products formed (hair, fur and nails). The arteriovenous anastomosis (providing the fine temperature regulation) was formed. Thus, in general, every contact surface between the organism and the air must perform a fundamentally new function.

By eliminating the lifting force, the supporting tissues must be given more tasks; the hollow skeletal bones were formed. Blood formation took place in the bone marrow instead of the liver and spleen.

Due to the new biosphere being transformed in principle, respectively, the locomotor system, cartilage substance, conductors, synovial membranes, muscle tone were degraded, and so on.

During life in water, the gills exerted the nndnlng function and participated in the organism's regulation of ion management. This was made possible by the fact that the seawater contained all the ions necessary for the maintenance of life and the essential trace elements. Due to the change in the biosphere, this role accrued to the two hormones in the parathyroid glands, namely the parathyroid hormone and litoralon, as well as the adrenal cortex hormone aldosterone. The parathyroid hormone primarily regulates calcium and magnesium regulation. Aldosterone controls the potassium and sodium regulation and litoralon primarily controls the regulation of the essential trace elements, including silicon fluorine and copper. Both hormones from the parathyroid glands participate in the regulation of the phosphate and sulphate ions. The serum citrate ion concentration is satisfied by the pareth hormone and lowered by litoralone.

During the transition from water to land, the immediate uptake of the necessary ions from the biosphere via the biomembrane was made impossible and the only possibility for the higher organisms to maintain ion balance was the uptake and depletion of nutrients (hormonal regulation of intestinal lining and intestinal lining). ).

As a ion reservoir, the bone skeleton required for life on land was trained with a stable structure. litoralon significantly affects the silicon content of the blood and plays a crucial role in its regulation (the silicon content of the blood increases during a litoralone treatment of rats and hares by more than 100% within 50 days). This fact draws close attention to the very important physiological role of silicon in glycomucocyte protein synthesis and the provision of its physiological functions (silicon in this context is similar to sulphate ions). i Probably as a result of the litoralone research the silicic acid and its complex will be moved from its hitherto 40 45 50 40 <2 '? 210027-l + insignificant location and have a prominent physiological role (build-up of bradytrophic and elastic tissues, regulation of bone formation- ningen etc.).

Otherwise, the silicon content of human blood corresponds to the silicon content in seawater (5-4 ppm) in which life first developed.

The evolution of life on land caused the organisms, which adapted to this way of life, the greatest stress effect that has ever occurred in the history of life on earth. This stress effect is exerted on the organisms living on land - even after the emergence of mammals and adaptation to life on land - to a much greater extent than on the organisms that live in the water (eg climatic effects, electromagnetic radiation, etc.). Most researchers have explained the extinction and emergence of certain animal species (for example, from the extinction of reptiles about 60 million years ago in the recent Cretaceous and the behavior of mammals) by changing the composition and quantity of solar radiation, regardless of whether it is caused by polar migration. or change in the activity of the sun. On the other hand, the parathyroid glands and their degree of development (the parathyroid glands secrete the only natural substance that protects against radiation and aging) determined the survival of the higher animal world on land and contributed to more resistant species becoming even more resistant to the exchanges. surface is exposed (seawater absorbs the ionizing radiation). It can be said with certainty that the intense development of the parathyroid glands in mammals made them more resistant to cosmic radiation.

Litoralon has a very important role, which at present can not appreciate, in defense against free radicals and browning of their formation, which free radicals are caused by the background radiation and are the main cause of aging. This effect is not only prophylactic, but damage that has already occurred of this kind is gradually restored, at least in part.

The population of the adaptation syndrome is insured for the higher organisms living on land by the Thalamus-hypothalamus-pituitary-adrenal cortex system. The litoral hormone from the parathyroid glands is an immediate component of the neuroendocrine adaptation system and its action gives rise to the stabilization and increase of the V resistance of the adaptation syndrome. It increases the body's ability to defend itself against harmful external influences. In this respect, it also has a pronounced effect on the higher nerve and brain activity and is also indispensable for the physiological function of the nerve system. 101 "50 w 7210027-4 111 It should be pointed out here that the method for the species 'survival in water differs in principle from that which applies to the species' survival on land. The development of the embryo in the body cavity and the challenge of the offspring through suckling developed under coercion circumstances, which occurred in connection with the transition to life on land, to the highest form of development (mammals currently living in the water have returned from land to sea), this ensures on land the safest maintenance of the offspring.The parathyroid glands exercise definite and expressive physiological effects on the gonads and reproductive organs and also on mammals' mammary glands.

In summary, it can be said that litoralon intensifies and in, strengthens the basic properties of the living tissues, raises the Ähñubner constant and thus the potential of the living state,. which constitutes the basis for development. Through intraoellular intensification of the inequilibrium and deformed state, litoralone has enabled the invasive spread of higher life forms over the entire surface of the earth. It has therapeutic value for any disease, the etiopathogenesis of which is causally related to the development of life on land and the adaptation to it. This is the explanation for the extensive therapeutic effect of litoralone.

In this context, a single example should be mentioned: Sjögren's syndrome- Wrmet is in the light of the above a typical lack of adaptability to the ability to live on land. The overall picture (which very rarely appears in all its symptoms, but whose partial or abortive form is so, much more common) consists of the following symptoms: Keraton conjunctivitis sicca Xerostomia, dysphagia Rhino-laringo-pharingitLs sicca Bronchitis sicca Pulmonary fibrosis Poliarthritis Scleroderma Akylia gastricemia, Vulvovaginitis sicca Alopecia Light dermatosis Teleangiectasia Sweat and sebaceous gland atrophy Calcification disorders of teeth and bones Numbness weakness, hypotension 40 45 55 40 721ÛÛ27'4 Purple Subfebrility Neurosis It is to be noted that all these symptoms were also etiologically pre-evolutionary. were inaccessible from a therapeutic standpoint.

Litoralonet therapeutically affects all of the above symptoms (both objectively and subjectively) very favorably if one, several or all of these symptoms occur.

Few ways similar to the theory developed above explain the origin or therapeutic treatment of numerous other syndromes. One can also recognize new syndromes and summarize many hitherto individually registered syndromes. It is thus clear that lithoralone deficiency is an important and in many cases a decisive cause of many diseases and not only in deficient cases can one expect a therapeutic effect of lithoralone. Like other hormones, litoralone acts on the stems and cells of the higher organisms (apocrine glands, mastocytes, lymphocytes, fibroblasts, osteocytes, chondrocytes, copper cells, etc.) by stimulating the adenyl cyclase enzyme by releasing adenosine monophosphate.

The difference between the hormone types of the parathyroid glands (parathyroid hormone and litoralone) can most easily be explained by the fact that parathyroid hormone exerts an effect of B-vitamin character and litoralone has an effect of vitamin A character.

It seems likely that hormones with a lithoralone effect are produced in the so-called eosinophil cells in the parathyroid glands.

Example 1. g of lyophilized and ground glandular powder was greased in a manner known per se (according to Aurbach): the powder was mixed with 25 ml of absolute acetone at room temperature, then filtration was carried out with a glass filter G. The well-aspirated glandular powder was then mixed with 25 ml of chloroform at room temperature and filtered through a G4 glass filter. The procedure was repeated. The aspirated substance was then suspended in 25 ml of absolute acetone at room temperature and filtration was carried out with a glass filter Gu. The glandular powder thus obtained was dried in vacuo at room temperature. The weight of the dry matter varied between 7 and 9 grams depending on the fat content of the starting material. g of defatted, anhydrous glandular powder was extracted with distilled water (5 times 100 ml) at room temperature for one hour. Between the extraction steps, the extract was separated from the glandular powder by means of a laboratory centrifuge with a high rotational speed. The extracts were stored before being worked up in a nitrogen atmosphere at a temperature of -5 ° C. The aqueous extracts were positive for ninhydrin, biuret and trichloroacetic acid.

The aqueous extracts were combined and added dropwise with a 50% by volume aqueous solution of trichloroacetic acid with constant shaking, until the final concentration of trichloroacetic acid was 15%. After standing at 5-5 ° C for one hour, the trichloroacetic acid precipitate was centrifuged off.

The upper layer consisted of a clear, pale yellow aqueous solution, which was biuret and trichloroacetic acid negative but ninhydrin positive.

The excess trichloroacetic acid was removed by series extraction at room temperature with an aqueous solution of ether (1-1, parts by volume of ether: 1 part by volume of water). In general, it is necessary to repeat the extraction 17-18 times in order to completely remove the trichloroacetic acid. In practice, the extraction is carried out 20-25 times.

After the last extraction, the aqueous phase must have a pH of 4.5-5. The aqueous phase was liberated from ether under a water jet pump vacuum at a temperature not exceeding 50 ° C. The ether elimination lasted for 6 hours and resulted in a reduction in volume (some of the aqueous phase was also distilled off). The ether elimination ~ was monitored by smelling the product.

The aqueous solution was then lyophilized and stored in a tightly closed vessel. The product is hygroscopic. The yield calculated on the total amount of glandular powder (before degreasing) was 10-15% (depending on the fat content).

Example 2. In the manner set forth in Example 1, an extract free from egg whites and precipitants was prepared from aqueous extracts of defatted parathyroid glands. 5 g of a trichloroethical inactive lyophilisate was filtered through a column of Sephadex G-15. The dimensions of the column were 4.3 times 144 cm. Water was used as eluent. The flow rate was 60 ml / hour. The fraction size was 15 ml. 50 ao 17 '7210027-4 Weight of the collected and lyophilized sample.

Number of tubes' Weight, mg 46 - 85 915 84 - 98 1126 99 - 118 999 119 - 136 1u16 157 - 154 24 155 - 1'70_ 3 185 - 198 19 221 - 250 2 258 - 265 15 279 - ess 1 mo fi alt a51a mg (90%) The main fraction with tube numbers 46 - 85 is with its content of 915 mg relatively rich in litoralone and has a pronounced physiological or therapeutic effect at a dosage of 5-10 micrograms / kg body weight. This is the substance obtained at the first two tips of the gel filtration. The fractions can be conveniently separated as follows: a) spectrophotometric absorption measurement at 280 nm bd spectrophotometric absorption measurement at 254 nm c) spectrophotometric absorption measurement at 560 nm. d) measurement of the electrical conductivity e) qualitative examination for chloride ions with silver nitrate f) pH measurement in the combined fractions belonging to the individual peaks g) examination with ninhydrin h) examination with the naked eye for opalescence and color (yellowish) .

Yield of main fraction 46-85 based on the amount of dry, non-defatted parathyroid powder was approximately 2-2.5%.

Example 5.

The procedure described in Example 2 was repeated with the change that the first two peaks were captured in separate main fractions. In this way 502 mg of main fraction I and 555 mg of the main fraction II were obtained from 5 g of egg white-free water extract.

Thereafter, main fraction II was subjected to renewed gel filtration. 456 mg of substance were taken up on the column and water was used as eluent. The dimensions of the column were 4.5 times 144 cm, the flow rate 50 ml / hour. The fraction size was 10 ml and the filling fl oïf ': Ä' consisted of "Sephadex",. 7210027-4 18 G-15.

In this way, the main fraction II could be divided into further peaks. The fractions belonging to the middle peak were combined.

Their weight after lyophilization was 257.5 mg. The yield calculated on non-defatted parathyroid powder was 0.6-0.8 7. Characteristic properties: yellow hygroscopic substance containing C, H, H, O, S and / or B. Characteristic groups: -SH, -COOH, -OH . In mass spectrographic examination, the following characteristic composition was obtained: Npozs- p U .It is assumed that a molecule has a side chain with a sulphanydryl content and is a purine derivative. The fractions contain additional structures. These fractions have physiological and therapeutic effects already at a dose of 1 microgram / kg body weight. Produk- å; _20 'z0_- 4o_' Éf._ten can be further purified by chromatography or otherwise. p Using a cellulase or ion exchange column, the polluting amino acids (lysine, glutamic acid) can be removed. In this way, a still contaminated lithoralone can be obtained in a yield of 0.5-0, h ä, which, however, is effective in a concentration even below 1 microgram / kg body weight. The yield is calculated on dry parathyroid powder.

In Example 4. The substance described in Example 4 was gel filtered on Sephadex G-15 (from Pharmncia, Uppsala) with a grain size of 90-420 .mu.m with decreasing technique, i.e. in the direction from top to bottom.

The elution diagram was measured at 280 nm extinction.

When determining the flow-through volume of liquid as a function of the extinction, litoralon was obtained as the second peak of the curve calculated from the beginning.

The characteristic gel filtration parameters were as follows: Amount of starting material: 5.4 g of substance (Example 1) Diameter of the column: 4.85 cm Content of the column 149 on Flow rate: 0.78 ml / min. ride: 10 min. Fractional sizes: 7.8 ml / test tube Top A Test tube 71-105 in "B" 104-128 "C" 129-v54 n ID u 1 55_1 62 "E" '1e5-189 "F" 190-247 "G "218-2? 7 The top B contained the active ingredient. 40 'BO 55 40 0“ Û 7210027 ~ 4 The volume of elution in 104 tubes was 814 ml. The total volume of the top B was 188 ml.

Between peaks D and E, the chlorine reaction is positive. The dry weight of the lyophilized peak B varied between 0.43 and 0.55 g, starting from 5 g of substance. "Identification by gel filtration: After the gel filtration on" SephadeiG§G-15, the substance appeared in the eluate at (0.67 É 0.02) x V0 if the determination was performed in aqueous solution by absorption at 280 nn.

V0 means the volume at which the salt occurs in the electron (positive chlorine reaction). The peak B was fractionated on a column ("Dowex" 50 x 2) at pH 1.8. 501 mg of substance in a 30% aqueous solution was applied to a 152 X 1 column ("Dowex" \ - 50 x 2), previously balanced with a formic acid-acetic acid blend of pH 1.8. The flow rate was 15 ml / h, the fraction size 4 ml and the fractions were determined at 280 nm. 9 elution peaks were obtained in this way: 2922 Test tube Weight, mg A 9 - 12 8.1 »B 13 - 17 18.4 'C 18 - 25 64.4 9D 94 - 29 5.2 E 50 - 56 ^ 7, 2 F 57 - 4? 0.5 G sz - so 5.1 H 51 ~ 57 7.5 I 58 - 80 6.8 146.5 (55.6%) The majority of the amount of active material obtained in fractions D, E and F was 7.9 mg, 2.6% of peak B.

The above fractions were applied to "Whatman" C) 5 mm paper 22 mm wide and electrophoresed at pH 1.8 with 1500 V for 90 minutes. The material load was 0.5 mg / 9 cm. The composition of the buffer was 40 ml of 98% formic acid and 160 ml of glacial acetic acid in distilled water to a total of 2 liters.

The active ingredient is ninhydrin positive and travels uniformly from the starting line towards the positive pole (5 in 90 minutes at 1500 V).

The product isolated by electrophoresis was eluted with water and the biological activity was tested. The yield of isolated product * calculated on a dry basis @ 1pu1ver was 0.01-0.0%.

The litoralone thus obtained can be considered as chemical, '55' v21ooz7-t of pure and its biological activity was proved by the following experiments: The weight of the herder gland in rats was considerably increased at a dosage of šaf / 120 g (subcutaneous administration). At the same dose, the blood sugar level was significantly reduced.

With oral administration of 7 micrograms (tablets) three times daily, the symptoms of rhinolaringo-pharingitis sicca were cured in humans. The amount of adenosine monophosphate excreted in the urine increased significantly.

Example 5.

Following the procedure described in Example 1, the loophilized final product was subjected to dialysis. 700 mg of substance dissolved in 55 ml of distilled water were dialyzed by means of a dialysis membrane (Kalle 44). The dialysis was carried out against water (? X 100 ml and then 15 x 150 ml) in such a way that the outer aqueous phase was replaced at 5.5 hour intervals. During the three intermediate nights, the dialysis time was 16 hours each time. The dialysis was carried out at a temperature of 5 ° C. Inside the membrane, 41 ml of the solution (non-dialyzed portion) was retained. Dry matter after lyophilization was $ 55 mg. The outer phase was evaporated in vacuo at 30 ° C and then subjected to lyophilization. Nan thus obtained 662 mg of dry matter.

Example 6.

The procedure described in Example 1 is repeated with 20 g of glandular powder (lyophilized, non-defatted). The aqueous solution liberated from egg white and ether was evaporated to one third of its original volume and then shaken 3 times with chloroform in a 1: 1 volume ratio. The aqueous phase was liberated in vacuo from chloroform. The chloroform-free aqueous solution was further evaporated in vacuo to 20 ml and added at room temperature with 5 parts by volume of acetone. The solution was allowed to stand on ice all night.

A yellow, oily phase separated from the almost colorless ncetone phase. The acetone solution was decanted off and the oil-like phase was dried in vacuo to constant weight. 1.1 g of a yellow resin-like concentrate (A) were thus obtained. The acetone solution was evaporated in vacuo to dryness to give 1 g of a resinous concentrate (B).

Claims (1)

Methods of preparing a preparation containing a previously undescribed hormone, here called litoralon, which hormone is produced by the parathyroid glands of mammals, has a mesenchymal activating effect, degrades the silicon content of the blood and has the following characteristics: (1 ) is soluble in water but insoluble in benzene, chloroform and carbon tetrachloride, (2) contains the chemical elements C, H, O, N and S, (5) has the following physiological effects: causes phosphaturia, has a radiation-protective effect and affects trace elements - the concentration in the blood, characterized by the drying of parathyroid glands, parathyroid tissue or cell cultures thereof and grinding, the product obtained 'is defatted in a manner known per se at room temperature, whereupon it is extracted with water, the aqueous extract is liberated from solids and further purified, precipitating proteins and polypeptides with trichloroacetic acid and removing excess precipitates to give at room temperature and in series extractions with ether, until the aqueous extract shows a pH of 4.5-5, whereupon the purified aqueous extract is lyophilized and / or subjected to division into different fractions by precipitation with acetone. , chromatography, gel filtration, electrophoresis and / or dialysis. BEFORE PUBLICATIONS: Germany 580,284 (30 h: 2/10), 1,193,641 (30 h: 2/10) Other publications: f Chemical abstracts 52 (1958): 12049 g, 6461 h; 62 (1965): 6745 g.