CA2201768A1 - Powdery biogenic preparation from ossified deer antlers and a food additive based on said biogenic preparation - Google Patents

Abstract

The biogenic preparation from ossified deer antlers appears as a disintegrated initial product having the particle size below 0.16 mm, a specific surface of about 100 sq.m/g, and a pore volume of about 3.0 cu.m/g.
The food additive contains as a biologically potent product the powdery biogenic preparation from ossified deer antlers.

Description

~r ~2~76~
,, POW~EKY DIO~NIC l~RE~ ION I~I~OM OSSI~IED DEER ~NTLERS

J~ND A E~OOD ~DD I '1'1 VE ~'\SED ON SA I L~ B I OGEN I C PREPI~R/~'I' I ON

Field of the Invention The invention relates in general to curative and nutritive biologically active products, and more specifically it concerns a powdery biogenic preparation from oss ified deer antlers and a food additive based on said biogenic preparation.
The lnvention can find application in producing medicinal agents, foodstuffs enriched with biologically potent substances, and can also be used in cosmetics, as well as in fodder production.

Background of the Invention Known iTI the art are animal-ori~in bio~enic '' ~ 22~7~

preparations successfully applicable in medical and veterinary practice, pantocrine and rantarine being the most widely recogni~ed preparations obtained from deer antlers.
~or instarlce, ~ r~locritle wllich l~ csson~ially ~l 10%
alcoholic extract of antlers of deer (that is, Siberian stag, Manchu~ian deer, and sika deer) contains a great deal of biologicalIy-active substances that provide for a broad range of indications for its application (SU, A, 10 195,049). Pantocrine has the following chemical composition:

org~tlic su~ arlccs 75 w~
inorganic substances 25 wt.%, wherein:
total nitrogen 8.4 wt.%
protein nitrogen 8.3 wt.%
nonprotein nitrogen0.l5 wt.%
proteins 52.7 wt.~
lipids 5.6 wt.%
mono- and disaccharides 15.5 wt .~
ash 25.0 wt.%.

In addition, pantocrine contains a great proportion of macroand microelements (Ca, Na, Ag, Cu, Ti, Al, Fe, Si, Mg, Pb, P) among which Fe dominates. A principal part of the active principle of pantocrine is the lipoid fraction, while the biogenous bases, that i.s, choline .

~20~ 7~

arld ethanolamine are lhe predominan~ p~ospllolipids.
Stu~ies of pantocrine demorl 9 trated a considerable proportion Or cllolesterol, steroids (estrone, testosterone, progcsterone, cholesterol ethers with oleic, stearic, and butyric acids).
It-is noteworthy that as little as one half of lipids is transferred from antlers to pantocrine, and a ratio between their fractions is somewhat different. In addition, deer antlers feature a higher content of phospholipids and sterol ethers. The ratio between the lipids of different classes in pantocrine varies depending on extracting conditions.
The preparation rantarine is a liquid extract (1:1) from antlers of males of reindeer obtained by repercolation on 40~ ethanol ~cf. a textbook "Rantarine"
by I.I.Brekhman, Moscow, Vneshtorgizdat PH, 1978 (in Russian). As is evidenced by studies performed, raindeer antlers differ from those of sika deer only in the amount of saccharides which is twice as large. (13.4 wt.~) in reindeer antlers. That is why rantorine and pantocrine are principally similar as to chemical composition, that of rantorine being as follows:

Essential amino acids, mg/l:

arginine 0.404 valine 0.823 ~2~ 7~

histidine 0.078 isoleucine 0.340 leucine 0.831 lysi~e 0.505 methionine 0.080 threonine 0.142 phenylalanine 0.61G
______________________________________________ Total 3.319 mg/l Elements, mcm/ml:
calcium 53.93 potassium 2588.4 sodium 3019.4 magnesium 34,5 iron 2.17 copper 1.73 zinc 0.65 manganese 0.02 nickel 0.02 chromium 0.02 Total 5701.2 mcm/ml The aforementioned animal-origin medicinal preparations have found widespread use as physiological 220~ 7~

. 5 stimulators, in disorders of the nervous system, dysmetabc)lisn), athcrosclcrosi.~:, hyt-otcr)siotl~ nrl(l se~u~ll impotence.
Both rantarine and pantocrine are dispensed not only as alcoho~ic extracts ~ut also as tablets, wherein said extract serves as t~e active principle.
The potency of antler extracts is to a great extent influenced by the preservation teehniques of antlers, as well as by the extraction procedure. It is during preservation of antlers and extraction of pantocrine therefrom that the valuable components of the initial product, that is, proteins, peptides, and blood elements are subject to destruction to a considerable extent. On the other hand, unpreserved antlers can be stored for as little as 6-lO hours after cutting-off, or within lO
days at minus 4OC.
Furthermore, the generally adopted antler extraction techniques fail to provide a complete isolation of pharmacologically valuable substances, otherwise speaking, all the preparations obtained from deer antlers known currently are depleted in ingredients compared with the initial product. Thus, it has been found that more than 9O~O of a total content of valuable and biologically-active antler components remains in the waste product o~ the production process, that is, in the residues.
~part from the aforesaid, the abovementioned ~ 22~176~
.

biogenic preparations possess a pronounced side effect, corlsisting in hemolytic and allergenic properties which is due to a considerable content of proteins and peptides, many of t~em being toxic. A large cholesterol S content remaining ;n the finishe(l product, is caus~tive of atherosclerosis, lipoid hyperplasia, and other diseases stemming from an excess amount of cholesterol in human organism.
Known in thc art are nutritional additives ~nutriceutics, naturopathic agents) containing biologically-active products in the form of concentrates of na~tural biologically-active substances or those identical thercto ol>~aincd frc)m prOCCSsinJ3 vca~et~t)le <~r animal raw stock, as well as by chemical or biotechnological methods using traditional, nontraditional, or special techniques and adapted for direct intake along with food or for being introduced into specialized food products with a view to enrichirlg the ration with individual nutrients and/or biologically active substances, or with their complex.
The aforementioned nutritional additives are produced in the form of extracts, fusions, balms, isolates, powders, dry and liquid concentrates, syrups, tonics, tablets, or capsules. The problem of elaborating and applying nutritional additives enhancing resistance of human organism to harmful effects of the surroundirlg medium becomes still more urgent in the regions with ~ 22~1768 ?~

unfavorable ecological situation.

Summary of the Invention It is an object of the present invention to provide a powdery biogenic preparation from a readily available animal raw material which is better balanced, as to its compos;tion, to human organism.
It is another object of the present invention to provide such a food additivc based on animal raw material that allows of sanitation of human organism and increasing its adaptogenic resources, as well as avoiding any side er~ect thel-eor.
The foregoing objects are ~ccomplished due to the provision of a novel powdery biogenic preparation from ossified deer antlers which consists of porous particles of a disintegrated initial product, the dispersity of said particles being largely below 0.25 mm, their speci flc surface about 100 sq.m/g, and a volume of pores of about 3.0 cu.m/g.
The proposed invention is instrumental in the provision of a biogenic preparation with a retained natural potency of the initial product and possessing a high specific adsorptive activity, antacid, and enveloping properties, as well as a broad range of pharmacological action.
A variant of protection afforded by the present ~ 22~176~

invention is a powdery biogenic preparation containing substantially as follows:
lipids 0.13-0.25 wt.%
essential amino acids and pcp~idcs, 5 not less than 0.80 mg/g cholesterol 0.10-0.20 mg/g phosphorus, not less than3.5 wt.%
calcium 15.0-20.0 wt.
iodine 0.02-0.04 wt.~
10 magnesium, not more than6800.0 mcg/g barium, not more than 1.30 mcgtg manganese, nor more than 3.5 mcg/~
tin 0.50 mcg/g copper O.S-20.0 mcg/g 15 iron 60-240 mc~/g zlnc 60-120 mcg/g lead, less than 0.35 mcg/g cadmium, less than O.OOlmcg/g cesium 4-1~ mcg/g 20 strontium S-10 mcg/g potassium 300-600 mcg/g.
The present invention is instrumental in producing a biogenic preparation from ossified antlers of raindeer, wherein macroand microelements are in a ratio and amounts most completely meeting the physiological demand of human organism, said preparation having in this respect high adaptogenic activity, as well as ~ 220176~

immunostimulating and immunomodeling properties.
The herein-proposed blogenic preparation contributes, due to its composition, to a better act;vity of the cardiac muscles and promotes blood coagulation processes.
Another subject-matter of protection is a food additive containing a biologically potent active product which, according to the invention, ;s in fact a powdery biogenic preparation from ossified deer antlers, tlle porous particles of said preparation featuring a dispersity principally below 0.25 mm, a specific surface of about 100 sq.m/g, and a pore volume of about 3.0 cu.m/g; it is expedient, according to the invention that said biogenic preparation be principally of the following composition:

lipids 0.13-0.25 wt.%
essential amino acids and peptides, not less than 0.80 mg/g cholesterol 0.10-0.20 mg~g phosphorus, not less than 3.5 wt.%
calcium 15.0-20.0 wt.%
iodine 0.02-0.04 wt.%
magnesium, not more than G800 mcg/g barium, not more than 31.30 mcg/g manganese, nor more than 3.5 mcg/g tin 0.50 mcg/g copper 0 5-20.0 m~K/K

-~ ~ 2 ~

-- 1 o iron 60-240 mcg/g zinc 60-120 mcg/g lead, Ics~q than 0.~5 mc~,/p cadmium, less than O.OOln~c~/~
S cesium 4-12 mcg/g strontium 5-10 mc~/g pota~sium 300-600 mcg/g.

The hereln-proposed food additive makes possible both prevention and treatment of the aforementioned diseases. Thus, said food additive is capable of increasing human adaptogenic resources, normalizing the metabolic effect, enhancing immunobiologic potencies and defenses of human organism, and stimulating taking heavy metals and toxins O-lt of human organism.
Further objects and many of the attendant advantages of the present invention will be readily appreciated as the same becomes better understood by reference to the following detailed description of a novel powdery biogenic preparation, a food additive based on said preparation, experimental and clinical trials of the proposed biogenic preparation and of said food additive based thereon.

Description of a Preferred Embodiment The herein-proposed novel biogenic preparation appearing as a powdery product wherein approximately 80~

~201 76~

of porous particles thereof feature a dispersity below ().25 mm, ~ sE~cciric ~urr~lcc cr ~bout 100 sq.m/g, nn(l n porc v~lumc ~f nb~ 3.() cu.m/~, hns bcen ~roduccd fr~m ossified deer antlers, that is, Siberian stag, Manchurian deer, sika deer, reindeer, moose, and European stag.
The novel biogenic preparation appears MS
grey-colored loose powder having inclusions of light-brown particles, has a specific nonputrescent odor, and is virtually insoluble in water and organic solvents..
The weight percent of volatiles therein is as low as 13.0, and that of total ash is as low as 49.5.
Biological potency of the proposed preparation stems from the initial product.
A sample analysis of a batch of the proposed powdery biogenic preparation carried out by thin-layer, gas~liquid, highefficiency chromatography, UF-spectrophotometry, elementary analysis for C, H, N, S, and I, as well as chromatomass-spectrometry, polarimetry, titrometry, and gravimetry has found the following chemical composition of said preparation:
Carbon 20.0 wt.
Nitrogen 6.5 wt.
Sulfur ~ 0.1 wt.Y
Hydrogen 3.5 wt.Y
Phosphorus 4.2 wt.Yo Iodine 0.012wt.%

22017~

Calcium 174,940 mg/kg . Sodium 4,260 mg/kg = Potassium 582 mg/k~
Magne~ium 6,643 mg/kg Iron 77 mg/kg Zinc 95.5 mg/kg Copper 0.~6 mg/kg Manganese 3.5 m~/kg Chromium 0.22 mg/kg Nickel 0.~2 mg/kg Cobalt 0.04 mg/kg Lithium 0.9 mg/kg Rubidium 3.4 mg/kg Cesium 4.4 m~/k~
Barium 31.3 mg/kg Strontium 8.6 mg/kg Molybdenum 1.6 mg/kg Lead 0.32 mg/kg Cadmium < O.OOlmg/kg Arsenic 0.15 mg/kg Mercury ~ 0.005mg/kg Tin 0.50 mg/kg.

It is worth noting that analyses of the various samples of the proposed preparation may give slightly different results.
The herein-proposed powdery biogenic preparation 220~7~

l3 -from ossified reindeer antlers contains principally as ~ollows lipids 0.13-0.25 wt.
essential amino acids and peptides, not less than 0.80 mg/g cholesterol 0.10-0.20 mg/g phosphorus, not less than3.5 wt.%
calcium 15.0-20.0 wt.~
iodine 0.02-0.04 wt.%
magnesium, not more than6800 mcg/g barium, not more than31.30 mcg/g manganese, nor more than3.5 mcg/g tin 0.50 mcg/g copper 0.5-20.0 mcg/g iron 60-240 mcg/g zinc 60-120 mcg/g lead, less than 0.35 mcg/g cadmium, less than O.OOlmcg/g cesium 4-12 mcg/g strontium 5-10 mcg/g potassium 300-600 mcg/g.
The fact that the proposed powdery biogenic preparatlon from reindeer antlers contalns a combination of a large amount of biologically potent compounds and micro- and macroelements in a physiological ratio optimum for human organism, as well as its specific adsorptive activity are responsive for its abillty to ~ 22ol 7~8 regulate metabolic and redox processes, hemopoiesis, functions of endocrine glands. The preparation is also capable of increasin~ immunological status of the organism, stimulating the growth and development of the S bone and muscular tissues, and bringlng heavy metals and toxins out of human organism.
Complex studies into the chemical composition and physical properties of the powdery biogenic preparation from ossified deer antlers, as well as its preclinical 1~ trials have demonstrated that a great amount of active elements, such as calclum an~ magnesium, and barium which are the principal components of the osseous tissue, stimulates its growth and development, and also provides for a better activity of the cardiac muscles lS and promotes blood coagulation processes.
The proposed preparation features the content of steroid compounds, including those producing hormonal effect (estrogens, androgens, progesterone) one order of magnitude lower than the known preparations from deer antlers; furthermore, the proposed preparation contains the amount of cholesterol causative of atherosclerosis, lipoid hyperplasia, and other diseases, ten to fifteen times lower than the known preparations.
For instance, the iodine content in the form of iodated derivatives of amino acid tyrosine (tyronine, thyroxin, 3,5-diiodotyrosine, 3,5,3'-triiodotyronine, 3,3'-diiodotyronine) in an amount excecding 0.02 wt.

22~1 76g -lrcs ~ rlo~ rlct ivity ~.r ~llc crl(lOcri~lc ~yC;l~nl ~"
human organism, includin~ the thyroid glarld which ~ 1l 1 l u~rl~ l o ~ )o l i ,YI" .
. A great conterll ol mangarlese is imporlant l'or S activating the redox processes in human organism.
The proposed preparation contains a little amount of proteins and peptides many of which may be toxic or possess hemolytic and allergenic properties. In addition, it contain~ a great proportion of phosphorus which is extremely necessary for promoting the growth of the bone tissue and for normal functioning of the nervous system, as well as is indicated in vascular hypotension, rachitis, and diatheses.
The herein-proposed biogenic preparation from = 15 ossified deer antlers possesses antacid, adsorptive, and enveloping properties and exhibits also an acid-basic capacity which is important when using the preparation in treating peptic ulcer of the stomach and duodenum, gastritis, heartburn, gastrointestinal pain, and other gastro- and enteropathies, in which reduced acidity and proteolytic activity of the gastric juice is indicated.
The amount of heavy toxic metals (lead, cadmium, mercury, molybdenum, cobalt, and other) is within permissible ranges which do not exceed the established norms that rule out accumulation of toxic metals in human organism.
The evidences we h~ve obtained are indicative of ~ 22~7~

-- 1 ~

the fact that there is no substantial difference, as to chemical composition, between the powdery biogenic preparations produced from ossified antlers of deers of different species, that is, Siberian stag, Manchurian deer, sika deer, or reindeer.
The attained physicochemieal and organoleptic quality characteristics of the proposed powdery biogenic preparation from ossified deer antlers, including microbiological chnracteristics and content of heavy metals, provoke no objections from the hygienic standpoints.
Pharmacological and toxicological studies of the proposed powdery biogenic preparation from ossified deer antler~ have been performed on animals of different species, i.e., albino mice, rabbits, guinea pigs, using different doses of the preparation, the trialed ones inclusive. It has been found that the proposed preparatlon exhibits no acute toxicity: the doses 15-210 times (for mice) and 9-90 times (for rabbits) higher than the recommended produce no toxic effect. Nor have its toxic properties been detected in chronic experiment on rabbits administered 160 mg/kg of the preparation for a month. No changes have been observed in: weight of animals, relative weight of their viscera, a number of biochemical characteristics (total protein, albumin, globulin, creatinine, glucose), morphological characteristics of the internal organs, and ~' 22017~8 hematological characteristics of the peripheral blood~
Chronlc experiment on mice given daily doses of the preparation, according to the invention, in an amount of 225 mg/kg and 1225 mg/kg has detected no changes in the S relative weight of the internal organs and in their morphology. The proposed powdery blogenic preparation produces no local lrritant effect. No slgns of allerglc reaction have been observed when studying an allergenic action of the preparation in a dose 70 times the recommended one. The proposed prcp~ratiorl wcre administered to guinea pigs orally in a dosc Or l g/k~
three times a day; in 21 days a skin reaction was observed after applying an electuary of the proposed preparation to skin. In addition, the preparation is not cytogenetically active when administered in doses of 1300 and 7800 mg/kg (the dose six times the recommended one). When given in a dose of 1300 mg/kg the preparation produces an antimutagenic action.
Studies into the pharmacological properties of the proposed powdery biogenic preparation have detected as follows: the preparation reduces arterial blood pressure (by 13% on the average~ in rabbits, stimulates the development of the genitalia (experiments were carried out on sexually immature albino mice given the preparation in different doses, the best result being obtained with a dose of a 5~ physiological saline).
There are noticed an increase in the relative weight o~

~ 220176~
- l$ -the genitalia, a higher RNA content of the prostate epithelium (in the cytoplasm), and hypertrophy of the secretory elements, and hypertrophy of the cells of the seminal vesicles. The powdery biogenic preparation from ossified deer antlers possesses hepatoprotective and lipotroplc action, produces an effect on the immune system, and stimulates the lymphatic folllcles of the spleen. There is also established an effect produced by the proposed preparation on the intensity of oxygen absorptlon by cells in experiments in vltro. High doses o~ the preparation inhibit, and low doses stimulate oxygen absorption by cells. In addition, the proposed preparation has an adsorptive capacity and is therefore effective a~ains~ rood poisonin~.
The powdery biogenic preparation proposed herein has been studied in a number of specialized therapeutic institutes, where its positive effect has been established. Thus, it has been found in the Institute of experlmental endocrinology (Moscow, Russia) that the proposed preparation causes no negative shifts in the analyzed characteristics of the condition of the endocrine system and metabolism in test rats. The slight changes revealed are indicative of a general growth-stimulating effect produced by the preparation.
Presence of iodine in the preparation imparts a protective effect of the preparation on the thyroid gland in cases of its possible radiation injuries.

~ 220I 7~8 It has been found in the Central Institute of Traumatology and Orthopedics Research (Moscow, Russia) tl~at use Or the preparation on ~ simulation of bone fracture in experimental animals (rabbits) results in an 5 c~ r~ onlE)Ic~ Llorl ~r ~. c~ us~ ~0,lll"~
with control animals and causes no pathologic changes in the bone tissue. Use of the preparation in an exper~ment (bone-musclè lnjury) reduces inflammatory, dystrophic and microcirculatory changes, thus reducin~ secondary necrotizing of muscular tissue and promotes filling of the wound canal with maturing granulation tissue. The healing period of skin-muscle wounds is cut down nearly twofold compared with the control.
Experiments conducted in the Urology Research Institute (Moscow, Russia) enable one to infer t~at the preparation is capable of stimulating the functional capabilities of the kidneys and of the organs of the male reproductive system, it adds to resistance of tissues and cells to the effect of disturbing factors, in particular, microorganisms and their toxins.
Trialing of the proposed preparation on experimental tuberculosis ln inbred mice has demonstrated that the preparation produces a positive effect on tissue reactions in cases of tubcrculous inflammation in experimental animals infected with Mycobacterium tuberculosis lI37Rv, thus contributing to their survival. As evidenced by hislologic studies, llle proposed preparation is found to produce an inhibitory effect on the development of specific inflammation in the organs of experimental animals, starting from the initial stages of in~ection (8 days) and stimulates proliferation of lympl~ocytes not only in lhe immunocompetent organs, such as the spleen but also in target organs, that is, the lungs and the liver, affected with tuberculosis.
A positive effect of the preparation on tuberculous 10 inflammatioTI is especially conspicuous wllerl Usc(~ in complex thcrapy of tuberculosis along with arltibnCtC.rial dru~s, and i.~ most pronounced in the late period of the disease's (18 days). In this case a more complete resolution of tuberculous foci in the lungs, liver, and spleen.
Thus, the studies carried out have demonstrated thnt thc proposed prepnration possesses somc immunomodulating action which tells positively on resistance offered by test mice to tuberculosis infection.
Studies into the effect produced by the proposed preparat~on on the peripheral blood and bone marrow composition, erythrograms, and some of the biochemical characteristics of blood in rats give evidence that:
1. The general status (weight, behavioral reactions, rate of food and water consumption, the con~ition o~ thc gnstrointcstinal tract) in nnimals .

~ 220~7~$

given the preparation in doses of 7 and 13 mg differ in nothing from the similar characteristics in control animals.
2. The preparation administered to intact test rats S in the experimental doses causes no changes whatever in the amount of erythrocytes, hemoglobin, hematocrit value, and in the erythroid offshoot of the bone marrow.
3. Resistance of the peripheral blood erythrocytes to thc crfcct of u chemical hemolytic agcrlt incrcases irrespec~ive of ~he dose of the preparation.

4. When taken in both of the aforementioncd doscs the preparation contributes to a substantial increase in the amount of thrombocytes in the peripheral blood and to fibrino~en concentration in intact animals even concurrently with blood loss.

5. Intact animals administered the preparation exhibit a reduced absolute number of lymphoid elements in both peripheral blood and bone marrow.

6. When administered in both doses against a background of chronic blood loss the preparation contributes to restoration of the bone marrow erythroid offshoot and the red blood quotients under study. ~cid resistance of erythrocytes in blood-loss animals increases to a greater extent than in intact animals, this effect being dose-dependent.

7. Administration of the preparation to test animals against u back~ro~nd of blood 109s COntt'ib-ltC.q 22~7~

to restoration of the bone marrow granulocytic offshoot and of the peripheral blood polynuclear hemocytes.
Studies into the effect of the proposed biogenic preparation on experimental peptic ulcer of the stomach in test rats have demonstrated that the reparation processes in the animals given the proposed preparation proceed faster than in the control animals and thc reparation period is by 2 or 3 days shorter.
Studies into the effect of the proposed biogenic preparation on the development of a pathologic process in the pancreatic tissue in experimental pancreatitis have demonstrated as foilows.
1. The preparation is capable of inhibiting a pathologic process in the pancreatic tissue in early stage~s o~ t-~xpcrimt~n~l E~ncreatitis. This e~ret~l i.s especially clear cut in macroscopic examination (less pronolJncet3 e(lellla of tllc pnrlcrens and of ~hc surro-lrltJing tissues, the symptoms of adhesive peritonitis are nearly absent).
2. The preparation stimulates the development Or the process of reorganization in the pancreatic tissue which is evidenced by a more pronounced symptoms of same in the animals of the experimental group.
The preparation has been assessed for biological potency on a short-term culture of human epidermic cells, the following findings being obtained.
The preparatlon i.s but slightly soluble in WatCI', ~2~17~8 .

nontoxic to human epidermic cells whose adhesive properties increase with a reduction of the preparation clo~sc. Tllc l-rcl-nlntior~ ls n mnrkctlly prorlolln(c~l proliferative cffect, tllc proliferative responsc increasing wilh a rcduc~ion Or the preparatiorl dos~.
'I'hus, a dose of 0.l mcg/ml increases DNA synthesis threefold, a dose Or 1 mcg/rnl, twofold, and a dose of 5 mc~/ml, by 5~ ~erccnt, wllcre~ls the doses of J0 ~tl~1 ~n mcg/ml produce no effect whatever on the proliferative response. The dose-dependent effect of an increase in protein synthesis is the same as in the proliferative response; thus, a dose of 0.1 mcg/ml has been found to increase the protein synthesis 3.5 to 4 times.
When administered in doses of 0.1 to 0.001 mkg/ml the preparation produces no effect on expression of human histocompatibility anti~ens.
The proposed food additive has been tested, in the Nutr~tion Research Institute under the Academy of Medical Sciences of the Russian Federation (Moscow), for a number of toxic elements, calcium, and phosphorus.
Test samples were prepared in compliance with the norms prescribed in the USSR State Standard GOST 26927-86 entitled "Edible raw materials and foodstuffs.
Mineralization for determining toxic elements". Tests for lead, cadmium, zinc, copper, and calcium were conducted in keeping with the methodological instructions on atomic-adsorption techniques of ~2~1 7~

determining toxic elements in foodstuffs and edible raw materials. The maximum content of said toxic elements h~s been found to hc 0.001 mg/kg. The arsenic content of the ~roposcd roocl n(lditive was determined S colorimetrically to be 0.1 mg/kg. The inorganic phosphorus content was determined spectrophotometrically.
The examination findings are tabulated in Table 1.

Table 1.

Namc of elemeTIts Content, mg/kg 1. Lead 0.75 2. Cadmium 0.030 3. Arsenic 0.11 4. Zinc 89.1 S. Copper 2.23 6. Phosphorus, % 9.7 7. Calcium, ~ 23.0 The proposed food additive has been trialed in the clinic of dietotherapy of the Nutrition Rescarch Institute under the Academy of Medical Sciences of the Russian-Federation in the controlled hospital conditions on patients with cardiovascular pathology arl(l ~20176~

-- 2~ --Cl)tlCOmi~lrlt U(Jl~oSi ty, USlrlg U ~)I'Oa(J L'U~lgO 01' biochemical tests ch~racteristic of the lipid, carbohydrate, and protein metabolism, the immunological status and the condition of patients' antioxidant ~ 5 pro~ection. Tllc rln~litl~ bt~lirlc~ cn~ble onc to ~t~tc ~is follows.
- The answers given to a questionnaire by the patients sufferin~ from hypertensive disease with concomitant adiposity demonstrate that ingestion Or thc proposed food additive concurrently with reduced-caloricity hyl)osodiulll urlti-n~hcrosclc~osis ~Jict contributes to suppression of the sensatiorl of hunger in a ma~ority of patients. In this case, no symptoms of intolerance to said nutriceutic nor allergic reactions are observed.
- The degree of body weight loss expressed in percent of the body weight prior to the treatment and attained in the course of dietotherapy using the Ar ration and the proposed food additive, exceeds by S0 that attained when using the diet alone, which is seemlngly due to a reduced sensation of hunger the resultant psychological comfort.
- Enrichment of the hypocaloric hyposodium diet with the proposed nutriceutic promotes the hypotensive effect of the ration, which is pronounced to the greatest extent with respect to systolic arterial prcssure.

-~ 220~ 7~
. .

- Use of the ~ood additive under trial contributes to a considerably reduced abdominal discomfort in patients suffering from biliary and colic dyskinesia. In the course of treatment these patients exhibited reduced S meteorism, dyspeptic phenomena, and abdominal pain.
- The most substantial positive changes in the biochemIcal characteristics, especially pronounced in patients with concomitant hyperlipoproteinemia and attained due to enrlchment of the antiatherosclerosis diet with the proposed food additive, are observed with respect to cholesterol whose level in the blood serum is reduced about twofold compared with the control.
- Incorporation of the proposed food additive in the Ar diet renders a positive effect on the carbohydrate and protein metabolism, the functional state of the liver, and the characteristics of the blood coagulation and anticoagulation systems.
- An analysis of the changes in characteristics indicative of the condition of the system "peroxidation of liplds" - antioxidant protection - enables one to infer that the proposed food additive produces a favorable effect on the antioxidant status of the organism and on the metabolism of lipids, thus promoting a reliable reduction in the concentration of the primary and secondary products of peroxidation of lipids in the course of treatment.
- It is due to the use of the dietotherapy 22~1 76~

incorporating the proposed food additive that an immunomodulating effect of s~id nutriceutic ;s revealed, which manifests itself by a reliable increase in production of immunoglobulins A and C, within the norm limits, by about one-third compared with their level before treatment.
The recommended daily dose of the proposed food additive is 400 ml/day before taking meals (breakfast or dinner) and is to be washed down with 100 ml water. A
total duration of the treatment course i8 one month. A
treatment course can be repeated two or three times a year.
According to evidence of functional, histologic, and morphometric examinations, the proposed biogenic preparation produces an effect of two kinds when used iTl an acute infectious morbid process. On the one hand, it prevents tissue destruction in case of an acute purulent inflammation, and on the other hand, it stimulates the system of Iymphomacrophagocytes, which is morphologically demonstrated in an increased amount of lymphoid follicles and a larger area thereof. A lesser extent of tissue destruction results in a better functional intactness of the kldneys and in restoration of all characteristics deranged due to the inflammatory process.
~ The experiments conducted allow one to infer that the proposed preparation is capable of stimulating the ~201 768 functional capabilities of the kidneys and the male reproductive organs. Moreover, resistance of tissues and cells to the effect of disturblng factors, in particular, microbial agents and their toxins, is increased due to nonspecific stimulation of the defense mechanisms, whereby the function of the affected organs ls lmpaired to a less extent and normalizes more quic~ly.
. A total of 20 patients (8 males and 12 females) aged from 36 to 58 and sufFering from hypcrtensive disease of stage I-II witll concomitant s~age I-TI
adiposity were observed under controlled hospital conditions. The patients of the control group (10 persons) were given a caloricity-reduced hyposodium, hypolipidemic antiatherosclerosis diet Ar, an energy (caloric) value of the ration being 1500 kcal. 10 patients of the experimental group were instituted the same diet for the same period, and additionally they took 400 ml of the propolsed preparation daily in the morning or the day-time before meals and washed the preparation with 100 ml water.
The patients of the experimental and control groups were identical as to the nature and degree of severity of the principal disease (that is, stage I-II
hypertensive disease), the concomitant pathology (hyperlipoproteinemia of a primary and secondary y,CllCS jS, I)i I i~lry ~In(l col ic (Iy~kine~ , (lel'ollllill"

~2~ 76~

spondylosis and osteochondrosis of the cervicothoracic and lumbar spine), and the sex and a~e characteristics.
An excess body weight not exceeding 30% was noted in 25%
of the patients in both experimental and control groups.
It is no mere chance that the patients with stage I-II hypertensive disease and concomitant adiposity, as well as those with concomitant hyperlipoproteinemia were selected as the object of observation in the aforesaid clinical trials. The presence of absorptive and adaptogenic properties in the food additive under trial is directly linked with the pathogenetic mechanisms of affection of the cardiovascular system in the patients under observation and, in particular, with lipoid dysmetabolism, modified immune reactions, and disturbed interrelations in the system "peroxidation of lipids -antioxidant organism protection".
Characteristics of the Ar diet used.
The chemical composition and energy (calorific) value of the Ar diet used are presented in Table 2.

Table 2 Chemical composltion and energy value of Ar diet Characteristics Quantity Proteins, g 75.0 including animal ones 45.0 22~17~8 Fats, g 70.0 including vegetable ones 25.0 Carbohydrates, g 1~0.0 including simple ones 15.0 Caloricity, kcal 1690.0 Vitamins:
Bl, mg 0.75 B2, m~ 1.46 B6, mg 1.57 C, mg 92.0 PP, mg niac.eq. 11.6 ~, mg toc.eq. 3.25 A, mcg ret.eq. 200.0 beta-carotin, mg 4.3 Mineral substances, g:
sodium 1.8 potassium 4.0 calcium 0.8 magnesium 0.6 phosphorus 1.2 Table 3 Generalized amounts ~f losses due to heat treatment Food Bl B2 C PP A beta- Ca Mg P
additive carotin ~ ~2017~

Percent of loss 28 20 ~0 20 40 20 12 13 19 Hyposodium caloricity-reduced antiatherosclerosis diet Ar is featured by a reduced amount of animal fats and oligosaccharides, as well as by a physiological norm of protein, restriction as to cholesterol-containing foods and extractive substances, and enric~ling the ration with foods containing lipotropic substances, polyunsaturated fatty acids, and edible fibers. The mineral composition is balanced adequately to the pathogenetic essence of the coronary heart disease and t~e hypertensive disease.
A complex examination of the patients included studying the changes in the objective signs of the disease, body weight, arterial pressure, a biochemical examination performed with the aid of the model "Spectrum" analyzer available from Abbot Co., USA, using a standard set of reagents and a standard programme.
Lipoid metabolism was assessed against a concentration of total cholesterol and triglycerides in blood.
In order to assess protein metabolism, total protein of~ the blood serum and uric acid were determined, while carbohydrate metaboli~m was asscsse(l ~201 76~

by determining glucose.
The functional capability of the liver was assessed against the content of total bilirubin, AST and ALT in blood serum. The condition of the coagulation and anticoa~ulation blood systems was judged against the ~uick's prothrombin consumption index, fibrinogen was determined by the Rutberg's technlque, and blood fibrinolytic actlvity, by the Kowalski, Kopek, and Niverski technique.
The condition of the system of antioxidant protection and of peroxidation of lipids was assessed by measuring the following characteristics in erythrocyte hemolysates:
- Activity of superoxide dismutase (SOD) was determined using a classical technique according to Beauchamp C., Fridovich J., 1971.
- Activity of glutathione peroxidase (GP) was assessed using the method of Mille G., 1959 in modification for the automatic analyzer "Clinicon-Korona" available from the firm LKB, Sweden.
- Activity of glutathione reductase (GR) was determined on the basis of a classical technique of Tilbotson J.A. et al., 1971 in modification for the automatic analyzer "Clinicon-Korona" available from the firm LKB, Sweden.
- Activity of catalase was determined by the technique developed and modified for the automatic 2201 7~8 analyzer "Clinicon-Korona".
- Content of diene conjugates was determined by the method of I.D.Sta~nay~ and T.G.Garishvili, 1977 in modification for erythrocytes.
5The following components ofnonenzymatic antioxidant protection were determined:
Uro~ In 1)l ood tlOrl~lll,U~l I n8 1~1 lln~lrl~U
sp~ctrophotometric technique.
- Corlc~rl~ra~lorl or vit~mln~ A ~ln(l E ln blood ~erum ~0 w~9 (~ctormincd by ~he fluorimetric technique o~
R.I.Cherliauskene, 1984, that of vitamin C, by titration.
Concentration of immunoglobulins A, G, and M and of the components of complements C3 and C4 was determined by an immunochemical technique, usin~ the model "ICS-II"
analyzer available from Beckman, USA.
All examinations were conducted dynamically so as to assess the effects of the performed alimentary intervention.
No symptoms of intolerance of the proposed food - additive nor any allergic reactions were noticed in the course of day-to-day clinical observation of the patients during clinical trials.
Contrariwise, the patients suffering from biliary and colic dyskinesia who were given the proposed food additive, exhibited reduced meteorism, dyspeptic phenomena, and abdominal pain. A majority of the ~2~17~8 patients noted a reduced sensation of hunger.
With a view to revealing a lipolytic effect of the food additive under trial, there was performed a comparatlve analysis of the degree of body weight loss in the patients given the proposed food additive concurrently with the antiatherosclerosis diet and in the patients of the control group who were instltuted the basic diet alone. A total body weight loss in the patients administered the proposed nutriceutic was found to exceed that in the control patients, thut is, ar average loss of weight in the experimental group was 6.4~ of its level prior to treatment, whereas that in the control group equalled 5.2%. An average daily loss of body weight in the patients of the experimental and control groups was 305 g and 247 g, respectively.
Adding the proposed food additive to the ration promoted the hypotensive effect of said ration. Thus, for instance, in the course of the dietotherapy the level of the systolic arterial pressure in the patients of the experimental and control groups was reduced by 31% and 24~, and that of the diastolic pressure, by 19 and 15%, respectively, compared with the figures before the treatment.
When studying the effect of the proposed food additive on biochemical indices, attention is attracted by the pronounced changes in the level of total cholesterol in blood serum. Thus, in the course of the :, . ;

22~1 7~
.

dietotherapy a hypocholesterolemic effect in the patients of thc expelitrlcrltul Kroup was ~ual to 2()% ol' the initial level, while that in the patients of the control group was 9%. The level of triglycerides in S blood serum remalned virtually unaffected as a result of treatment, being in the limits of normal variation in the patients of both groups. A hypoglycemic ef~ect in said patients was found to be appro~imately the same as well.
Chan~es in the level Or total protein itl bloo~
serum were insignificant, and the blood uric acid content reduced rellably in the patients of both groups approximately at the same rate. There was also noticed a considerable reduction the blood level of AST and ALT, 15 as well a moderate decrease in the total bilirubin content in the patients ~iven the proposed food additives concurrently with the Ar diet (see Table 4).
Reduction of the prothrombin consumption index in the course of dietotherapy was twice as more pronounced in the patlents of the experimental group, whereas a favorable change in the fibrinogen level and fibrinolytic activity of blood was the same in the patients.of both groups.
When assessing the effect of the proposed food additive on provision with vitamins there was established a notlceable increase in the concentration of vltamlns A and E in the blood of t~le paticrlts bcinK

~201 76~

treated, whereas no such an effect was observed in the patlents of the control group (see Table S).
Use of the proposed food additive concurrently with the reduced Ar diet contributed also to improvement of S some characteristics and activation of the enzymatic antioxidant protection. Thus, for example, in the cou~se of treatment the patients of the experimental group sufferin~ from hypertensive disease and adiposity exhibited a reliable increase in the level of glutathione peroxidase and superoxide dismutase of erythrocytes, whereas such an increase was pronounced to a much less extent in the patients of the control g~oup.
The degree of reduction of the products of peroxidation of lipids (diene conjugates and malonic dialdehyde) in the course of the dietotherapy with the use of the proposed food additive was twice as high in the patients of the experimental group comparing with the control. A mar~ed antioxidant effect of the nutriceutic under trial might be, in partlcular, due to the presence of some microelements therein, which possess an antiperoxidant action, such as chromium.
Use of the proposed food additive as part of the antiatherosclerosis diet was found to produce a definite immunomodulatin~ effect. As seen f~om Table 6, no perceptible changes in the system of humoral immunity were observed in the patients of the control group given the basis diet. On the other hand, enriching the Ar diet 22~ 768 s with the proposed food additive contributed to a reliable increase in the blood concentration of immuno~lo~>ul 1 n~ ~ ~nd C .

Table 4 Changes in the clinicobiochemical characteristics resultant from the effect of the administration of the proposed food additive concurrently with the Ar diet in the patients with hypertensive disease and cQncomitant adiposity Character- Experimental group Control group istics before after before after treatment treatment treatment treatment 15 Cholesterol, mmole/l 6.59+0.175.49+0.18t 6.11+0.22 5.50+0.26 Tri-gIycerides, mmole/l 1.37+0.141.48~0.13 1.81+0.26 1.42+0.18 20 Glucose, mmole/l 6.66+1.46.17+0.85 6.23+2.11 5.64+0.gS
Total 2201 76~

l 2 3 4 5 protein, g~l 81.9+2.58 77.8+2.94 ~79.2+5.83 76.0+4.39 Uric acid, mcmole/l 252+90 261+94 320+63 255+52 Bilirubin, mmole/l 14.3+1.43 9.88+0.88 24.2+3.51 19.9+2.11 AST, IU/l 24.G+3.39 17.8+1.64 26.5+3.24 22.8+3.22 ALT,IU/l 15.6+4.20 17.7+4.78 39.4+4.25 31.5+3.97 Creatinine, mcmole/l 64.9+3.39 70.4+3.61 59.7+4.96 62,4+3.55 Urea, mmole/l 6.98+0.29 4.91+0.34 6.63+0.34 5.64+0.29 Albumin, g/l 44.9+0.48 44.3+0.62 43.1+0.29 42.7+0.35 Globulin, g/l 36.8+1.34 33.4+1.48 36.5+0.32 34.2+0.33 Calcium, mmole/l 2.48+0.37 2.48+0.29 2.52+0.24 2.49+0.25 Phosphorus, mmole/l 0.74+0.05 0.92+0.06 0.88+0.05 0.87+0.07 Prothrombin, % 98.1+2.11 90.7+0.99 92.0+4.92 84.3+2.72 Fibrinogen, mg % 343.0~23 372.5+27 371.2+45 322~-42 ~20~ 768 Fibrinolysis time, min 268.9~14.2 216.7+12.5 lg6.4~26.1 187.5~29 Note. * - with p < 0.05.

Table 5 Changes in the characteristics of the system "peroxidation of lipids - antioxidant protection" under the effect of the proposed food additive concurrently with Ar diet in the patients with hypertensive disease and concomitant adiposity Character- Experimental group Control group istics before after before after treatment treatment treatment treatment Glutathione reductase, mcmole NADPH/min/ml crythr.2.7~+0.122.45+().06 2.28+0.04 2.3~+0.12 ~ ~2~1768 Glutathione peroxidase, mcmole S NADPH/min/ml erythr, 35.4+0.27 48.2+0.14 32.4+0.29 35.0+0. 15 Catalase, kU/ml erythr. 261+16.2 244+10.0 290+7.3 297+10.2 Superoxide dismutase, AU/ml erythr. 2347+53 3741+60 2487+74 2706+81 Vltamin A, 15 sera, mg % 14.3+1.05 18.8+0.94 15.7+0.83 16.2+1.12 Vitamin E, sera, mg ~ 0.73+0.04 0.97+0.05 0.80+0.03 0.82+0.06 Vitamin C, sera, mg % 0.60+0.01 0.88+0.02 0.68+0.03 0.74+0.04 SH-groups, sera, mmole/l 2.56+0.09 2.78+0.11 2.38+0.11 2.48+0.07 Malonic dialdehyde, 25 mmole/ml 3.84+0.0~ 2.50+0.08t 3.68+0.09 3.07+0.12 2~17~g Diene conjugates, nmole/ml erythr,3.58~0.10 2.32+0.06 3.27+0.14 2.69+0.12 Table 6 Changes in the characteristics of humoral immunity under the effect of the proposed food additive concur-rently with Ar diet in the patients with hypertensive disease and concomitant adiposity Character- Experimental group Control group istics, mg/100 ml before after before after treatment treatment treatment treatment Immuno-globulin A247+14 336+181 278+20 303+14 Immuno-globulin G905+53 1234+61 947+54 912+60 Immuno-globulin M257+12 278+14 236+10 241+12 ~'~ 220~76~

Complement ~3 component 90.7~3.6 8~.0+2.1 95.4~4.0 92.3~1.7 5 Complement C~
componcrlt 28.41-l.0~ 26.3t1.34 25.7+0.94 27,4+0.76 * _ p c o.OS

Thus, in-depth chemical studies and extensive preclinical trials have demonstrated that ossificd deer antlers are a natural accumulator of valuable useful chemical elements and biologically potent components that have been maturing for 7-8 months.
That is why to use deer antlers in the stage of IS their development, that ls, antlers of young deer, implles producing medicinal preparations (pantocrine, rantarine) and food additives from an immature product which is more toxic and less efficient.
The herein-proposed po~dery bio~enic preparation from ossified deer antlers and the food additive based on said preparation having the characteristics as specified by the present invention, provide for adrninis~crin~ to llu~ n or~lr~ l the pro(Jucl r~alr~ l)y 100% its natural composition.

Claims (4)

1. A powdery biogenic preparation from ossified deer antlers, consisting of porous particles of a disintegrated initial product, the dispersity of said particles being largely below 0.25 mm, their specific surface about 100 sq.m/g, and a pore volume of about 3.0 cu.m/g.

2. A powdery biogenic preparation as set forth in claim 1, containing principally the following components;
lipids 0.13-0.25 wt.%
essential amino acids and peptides, not less than 0.80 mg/g cholesterol 0.10-0.20 mg/g phosphorus, not less than 3.5 wt.%
calcium 15.0-20.0 wt.%
iodine 0.02-0.04 wt.%
magnesium, not more than 6800 mcg/g barium, not more than 31.30 mcg/g manganese, nor more than 3.5 mcg/g tin 0.50 mcg/g copper 0.5-20.0 mcg/g iron 60-240 mcg/g zinc 60-120 mcg/g lead, less than 0.35 mcg/g cadmium, less than 0.001 mcg/g cesium 4-12 mcg/g strontium 5-10 mcg/g potassium 300-600 mcg/g.

3. A food additive, comprising a powdery biogenic preparation from ossified deer antlers, the porous particles of said preparation featuring a dispersity principally below 0.25 mm, a specific surface of about 100 sq.m/g, and a pore volume of about 3.0 cu.m/g.

4. A food additive, wherein said biogenic preparation contains the following components;
lipids 0.13-0.25 wt.%
essential amino acids and peptides, not less than 0.80 mg/g cholesterol 0.10-0. 20 mg/g phosphorus, not less than 3.5 wt.%
calcium 15.0-20.0 wt.%
iodine 0.02-0.04 wt.%
magnesium, not more than 6800 mcg/g barium, not more than 31.30 mcg/g manganese, nor more than 3.5 mcg/g tin 0.50 mcg/g copper 0.5-20.0 mcg/g iron 60-240 mcg/g zinc 60-120 mcg/g lead, less than 0.35 mcg/g cadmium, less tha 0.001 mcg/g cesium 4-12 mcg/g strontium 5-10 mcg/g potassium 300-600 mcg/g.