`(43) Application published 3 Jun 1983
`
`{21} Application No 8706977
`
`(22) Date of filing 24 Mar 193?
`
`(30) Priority data
`(31) 8823228
`
`(32) 26Nov19B6
`
`(33) GB
`
`(51)
`
`INT CL‘
`A51K 31:’195 31fl35 31i7D 311715 35478
`
`(52) Domestic classification [Edition J]:
`A53 180 231 20X 20‘! 230 23X 233’ 26Y 303 30X
`30Y 401 402 403 406 «WY 421 423 426 42Y 481
`462 483 -{BY 502 503 5OY 550 55Y 576 57‘! 555
`SBY 596 59‘! 616 61Y JA
`U18 1328 A53
`
`
`
`(71) Applicant
`lnstltut Blologll Marya Dalnavostoclmogo Nauchrlogo
`Tsentra Altadernll Nauk SSSH
`
`(56) Documents cited
`Nona
`
`(Incorporated In USSR)
`
`Vladivostok. prospekt 100 list, Vladlvosloltu 159,
`Union at Soviet Socialist Republics
`
`(53) Field of search
`A58
`06E
`Selected US specifications from IPC sub-class
`A51K
`
`Nauohno-lssledovalelsky Instllul Sadovodstva
`Vinogradarstva I Vlnodella
`
`(|n¢mpgm.ad in ussn)
`
`Tbilisi. Union or Sovlat Socialist Republics
`
`Giaptarovich Ohekutishuiti
`lzratl llskovloh Brekhrnarl
`
`Alexandr Evgenlevich Bulanov
`Mira lvanovna Polozhentseva
`
`(72)
`
`_
`Inventors
`(74) A99” afldm Address hr Sam“
`Levan Alexandrovlch Mudzhlrl
`”'*""* 5‘ °‘°",‘
`_
`Gla Gennadlevlch Alkhazaslwlll
`f_:'n°g°:':‘:;’é;; g'L';"'°""»
`Elena llilnlchna Kalatozishvili
`
`
`(54) Inhibiting alcohol addiction
`
`(5?) Compositions {or inhibiting alcohol addiction contain: leukoanthooyans, catochins. flavanols, lignin, reducing sugars,
`pectin. free aminoaclds. organic acids. sterols. methylsiorols, dimethystorols. lignanes, lignano glycosidss. phenolic acids,
`phenol aldehydes and alkyllerulates. Alcoholic beverages containing these compositions are also disclosed.
`
`‘VW086LZ89
`
`USP|abs EXHIBIT 1005
`
`_;‘L-4 '
`
`‘-oval;
`"
`
`USPlabs EXHIBIT 1005 .
`
`
`
`21980~*E1
`
`Composition Inhibiting Pathological
`
`Addiction to Alcohol
`
`Field of the Invention
`
`The present invention relates to compositions for
`
`5
`
`inhibiting the development of
`
`a pathological addiction
`
`to alcohol, and to processes for their manufacture.
`
`Background of the Invention
`
`Known in the art is an alcoholic bitter liqueur
`
`comprising the following ingredients, kg/1.000
`
`10
`
`decalitres:
`
`hralia root
`
`Eleutherococcus extract
`
`schizandra (fresh berry)
`
`sugar
`
`15
`
`Natural honey
`
`Tint
`
`2.5
`
`20
`
`345
`
`200
`
`50
`
`25
`
`Aqueo—a1coho1ic liquid .
`
`the balance.
`
`9‘
`
`[See "Formulations of Liqueur-Vodka Products and
`
`Vodkas". "Legkaya i Pistschevaya Promyshlennost" (Light
`
`
`
`2
`
`and Food Industry} Publishing House. Moscow. 1981,
`
`13.133).
`
`_.L3"V4'.a._gI
`
`Also known in the art is an alcoholic bitter liqueur
`
`including the following ingredients. kg/1.000 decalitres:
`
`U:
`
`Eleutherococcus extract
`
`Schizandra (fresh berry)
`
`schizandra (seeds)
`
`Natural honey
`
`Tint
`
`200
`
`92
`
`0.6
`
`50
`
`30
`
`10
`
`Aqueo-alcoholic liquid
`
`the balance.
`
`{See “Formulations of
`
`liqueur-Vodka Products and
`
`Vodkas“, Light and Food Industry Publishing House.
`
`Moscow, 1981. p.206.)
`
`The prior art alcoholic liqueurs are produced by way
`
`of blending the starting components, a successive
`
`introduction.
`
`into the resulting blend. of an aqueo-
`
`alcoholic liquid. settling of
`
`the resulting mixture and
`
`filtration thereof.
`
`As it is seen from the above—specified formulations,
`
`20
`
`the latter contain a biologically active extract of
`
`Eleutherococcus which lowers the toxic effect of ethanol
`
`in a living organism.
`
`
`
`
`
`2
`
`However.
`
`low taste properaties of the prior art
`
`beverages caused by a limited content of aromatic
`
`compounds. as well as a "pharmaceutical" aftertaste due
`
`to the presence of Eleutherococcus extract, high costs
`
`5
`
`of
`
`the starting ingredient — Eleutherococcus extract —
`
`and limited availability of this plant do not enable a
`
`wide scale of consumption of these prior art beverages.
`
`The present
`
`invention resides in the provision of a
`
`composition inhibiting the development of a pathological
`
`10
`
`addiction to alcohol which. according to the present
`
`invention, comprises the following ingredients. mg/g:
`
`leukoanthocyanes
`
`catechins
`
`Elavanols
`
`15
`
`lignin
`
`reducing sugars
`
`pectin
`
`free amino acids
`
`organic acids
`
`20
`
`sterols
`
`methylsterols
`
`dimethylsterols
`
`lignans
`
`7
`
`lignan glycosides
`
`25
`
`phenolic acids
`
`219-270
`
`153-187
`
`81-99
`
`63-83
`
`216-264
`
`18-22
`
`27~33
`
`36-44
`
`4.5-5.5
`
`1.35-1.65
`
`1.93-2.42
`
`13.5-16.5
`
`9-11
`
`13.5-16.5
`
`
`
`phenolic aldehydes
`
`alkylferulates
`
`4.5-5.5
`
`4.5—5.5
`
`-2:"
`
`'
`
`The composition according to the present invention
`
`advantageously comprises a combination of compounds
`
`LJ1
`
`occurring in nature.
`
`The composition has a pronounced
`
`capability of affecting processes or ethanol metabolism
`
`without switching to unfavourable routes of
`
`the
`
`organism's utilization of ethanol: as a result.
`
`the
`
`process of formation of a physical dependence on alcohol
`
`10
`
`is delayed,
`
`the level of its consumption is lowered and
`
`certain alcoholic behaviour excesses disappear.
`
`Furthermore,
`
`the composition according to the present
`
`invention is not
`
`toxic and is safe upon prolonged
`
`consumption: it has positive organoleptic
`
`,_.. LJI
`
`characteristics and can be useful as a food additive to
`
`alcoholic and alcoho1—free beverages.
`
`In a preferred aspect,
`
`the present
`
`invention resides
`
`in that an alcoholic beverage comprising sugar, citric
`
`acid.
`
`a tint and an aqueous—alcoholic liquid. according
`
`20
`
`to the present
`
`invention also incorporates fruit alcohol
`
`and a composition of substances inhibiting the
`
`development of a pathological addiction to alcohol
`
`containing, mg/g:
`
`leukoanthocyanes — 219-270, catechins
`
`— 153—l87. flavanols — 81-99.
`
`lignin — 68-83,
`
`reducing
`
`35
`
`sugars - 216-264. pectin — 18-22. free amino acids
`
`4
`
`wfi.
`
`
`
`5
`
`-27-33. organic acids — 36-44. sterols - 4.5-5.5,
`
`methylsterols — 1.35-1.65, dimethylsterols — 1.98-2.42.
`
`lignans « 13.5-16.5.
`
`lignan glycosides - 9-11. phenolic
`
`acids - 13.5-16.5. phenol aldehydes — 4.5—Sl5.
`
`alkylferulates ~ 4.5-5.5.
`
`the ingredients being present
`
`in the following proportions. kg per 1.000 decalitre of
`
`the beverage:
`
`the above—specified composition
`
`473-493
`
`fruit alcohol 40°
`
`4,950—5,050
`
`10
`
`sugar
`
`citric acid
`
`tint
`
`95-105
`
`1.8-2.2
`
`28—32
`
`aqueous—a1coho1ic liquid
`
`the balance.
`
`The alcoholic beverage according to the present
`
`invention is capable of
`
`inhibiting a pathological
`
`addiction to alcohol and has high organoleptic
`
`properties — the tasting test of the beverage is not
`
`less than 9.1 points.
`
`This alcoholic beverage may be produced by a process
`
`20
`
`comprising blending of sugar. citric acid and a tint.
`
`followed by the addition.
`
`to the resulting blend. of an
`
`aqueo—a1coho1ic liquid. settling and filtration. wherein
`
`according to the present invention blended are 473—493
`
`
`
`
`
`kg of
`
`a composition inhibiting a pathologic addiction to
`
`alcohol, 4.950—S.0SO kg of
`
`a 40° fruit alcohol. 95-105
`
`kg of sugar. 1.8-2.2 kg of citric acid and 23-32 kg of
`
`tint:
`
`to the resulting blend the aqueous~alcoholic
`
`liquid is added in the amount required for the
`
`preparation of 1.000 decalitre of
`
`the beverage;
`
`prior
`
`to settling and filtration the final blend is subjected
`
`to a triple successive thermal
`
`treatment for 5-8 hours
`
`at a temperature of 70—80°C and to cooling to attain a
`
`10
`
`temperature within the range of from 0 to —10°C.
`
`Other subjects and advantages of the present
`
`invention will now be more fully apparent from the
`
`following detailed description of
`
`the alcoholic beverage
`
`and the process for producing same according to the
`
`present
`
`invention.
`
`The beverage according to the present invention
`
`contains, kg per 1,000 decalitre of
`
`the beverage:
`
`Composition inhibiting a pathological
`
`addiction to alcohol
`
`473-493
`
`20
`
`40° fruit alcohol such as pear alcohol,
`
`apple alcohol, plum alcohol.
`
`tangerine
`
`alcohol
`
`sugar
`
`4.9S0~S,050
`
`95-105
`
`ll‘
`
`
`
`
`
`citric acid
`
`tint
`
`1.8-2.2
`
`23-32
`
`an aqueo—a1coho1ic liquid
`
`the balance
`
`(comprising a mixture of water
`
`and ethanol
`
`in a desired ratio)
`
`The desired ratio of ethanol and water in this
`
`mixture depends on the final strength of
`
`the produced
`
`beverage.
`
`10
`
`15
`
`when the alcoholic beverage of the present
`
`invention
`
`is consumed.
`
`the process of
`
`the formation of a physical
`
`dependence on ethanol
`
`in the organism is inhibited.
`
`Furthermore. drinking of this beverage is not
`
`accompanied by such negative effects as a "hang~over"
`
`syndrome.
`
`The tasting test value of
`
`the above alcoholic
`
`beverage according to the present invention is not less
`
`than 9.1 points.
`
`An improvement
`
`in organoleptic
`
`properties is attained due to an increased content of
`
`aromatic substances.
`
`in particular esters of derivatives
`
`20
`
`of aromatic acids and higher alcohols. as well as due to
`
`interaction of aldehydes. ketones, acetals and
`
`alkylferulates being present
`
`in the beverage composition.
`
`An alcoholic beverage according to the present
`
`
`
`
`
`B
`
`\_JI
`
`10
`
`invention is produced in the following manner. There
`
`are blended:
`
`a composition inhibiting the development
`
`of a pathological addiction to alcohol. a fruit alcohol,
`
`sugar. citric acid, a tint. whereafter the resulting
`
`blend is added with an aqueo—ethano1ic liquid in an
`
`amount necessary to obtain the beverage of 40%
`
`strength.
`
`Then the resulting blend is subjected to a
`
`successive heat
`
`treatment for three times at a
`
`temperature of 70—80°C for a period of 5-8 hours.
`
`Thereafter.
`
`the blend is cooled for a period sufficient
`
`to acquire a temperature within the range of
`
`from 0 to
`
`-10°C.
`
`Then the thus—produced beverage is filtered.
`
`allowed to stand for 10 days at a temperature varied
`
`within the range of
`
`from 10 to 45°C and again filtered.
`
`I--‘ U!
`
`Production Example 1
`
`There are blended 483 kg of
`
`a composition
`
`containing. mg/g:
`
`leukoanthocyans — 245.0. catechins —
`
`180.0. flavanols — 90.0.
`
`lignin - 75.0.
`
`reducing sugars
`
`— 261.7. pectin — 20.0, free amino acids — 30.6, organic
`
`20
`
`acids — 39.0, sterols - 5.0. methylsterols — 1.5.
`
`dimethylsterols - 2.2.
`
`lignans — 15.0.
`
`lignan glycosides
`
`— 10.0. phenolic acids — 15.0. phenol aldehydes — 5.0,
`
`alkylferulates — 5.0. 40° pear alcohol
`
`— 4.990 kg. sugar
`
`— 105 kg. citric acid — 2.2 kg. tint — 32 kg
`
`
`
`
`
`9
`
`r.
`
`and an aqueo—ethanolic liquid in an amount required to
`
`obtain a blend following heat treatment at the
`
`temperature of 75°C for 6 hours. Thereafter the blend
`
`is cooled to a temperature of 0 to 2°C.
`
`Then the
`
`5
`
`beverage is filtered. settled for 10 days at a
`
`temperature within the range of from 1o to 15°C.
`
`The final product is filtered while being bottled.
`
`Production Example 2
`
`A beverage is produced by blending 473 kg of a
`
`10
`
`composition containing the following ingredients in the
`
`amounts specified hereinbelow. mg/g:
`
`leukoanthocyans —
`
`219, catechins — 153. flavanols — 81.
`
`lignin — 68.
`
`reducing sugars — 345.17. pectin - 16. free amino acids
`
`- 27. organic acids - 36. sterols — 4.5, methylsterols —
`
`15
`
`1.35. dimethylsterols — 1.98,
`
`lignans — 13.5,
`
`lignan
`
`glycosides — 9, phenolic acids — 13.5, phenol aldehydes
`
`- 4.5, alkylferulates — 4.5.
`
`It also contains. as indicated in Example 1. 4.950
`
`kg of a 40°C plum alcohol. 95 kg of sugar. 1.3 kg of
`
`20
`
`citric acid. 28 kg of a tint and an aqueo—ethanolic
`
`liquid in an amount sufficient to obtain a blend with
`
`'?
`
`the strength of 40 volt. Thereafter the blend is
`
`subjected to a triple successive heat treatment for 5
`
`
`
`10
`
`hours at the temperature of 80°C.
`
`Then the blend is
`
`cooled for a period of
`
`time sufficient to acquire a
`
`temperature of 0 to 1°C.
`
`Then the beverage is filtered,
`
`allowed to stand for 10 days at a temperature of 20 to
`
`22°C.
`
`The final product is again filtered when being
`
`bottled.
`
`Production Example 3
`
`There are blended 493 kg of
`
`a composition containing
`
`10
`
`the following ingredients in the amounts specified
`
`hereinbelow, mg/g:
`
`leukoanthocyans - 2?0. catechins —
`
`137, flavanols — 99,
`
`lignin — B3.
`
`reducing sugars —
`
`197.5, pectin - 22.
`
`free amino acids - 33. organic acids
`
`— 44, sterols - 5.5. methylsterols — 1.65,
`
`dimethylsterols — 2.42.
`
`lignans — 16.5.
`
`lignan
`
`glycosides — 11. phenolic acids — 16.5, phenol aidehydes
`
`— 5.5, alkylferulates — 5.5. a 40° tangerine alcohol
`
`—
`
`5.050 kg, sugar
`
`— 105 kg. citric acid — 2.2 Kg, tint —
`
`32 kg, an aqueo—a1coholic liquid in an amount sufficient
`
`20
`
`to obtain a blend of
`
`a 40 vo1.% strength. Thereafter
`
`the blend is subjected to a triple successive heat
`
`treatment for 5 hours at the temperature of 80°C.
`
`Then
`
`the blend is cooled for a period of
`
`time surficient for
`
`it to acquire a
`
`temperature of B—l0°C.
`
`Then the
`
`
`
`
`
`ll
`
`beverage is filtered, allowed to stand for 10 days at a
`
`temperature of
`
`from 40 to 45°C.
`
`The final product is again filtered when being
`
`bottled.
`
`Further objects and advantages of
`
`the present
`
`invention will now become more fully apparent from the
`
`following detailed description of a composition
`
`inhibiting the development of a pathological addiction
`
`to alcohol with reference to examples illustrating its
`
`10
`
`particular embodiments.
`
`15
`
`20
`
`The composition according to the present invention
`
`preferably contains, as leukoanthocyanes,
`
`leuko-
`
`dolphinidine.
`
`leukocyanidine and leukopelargonidine. As
`
`catechins it contains {—)epigallocatechin.
`
`(+)gallocatechin, {-}epicatechin, {+) catechin and
`
`{—)epicatechinga11ate.
`
`As flavanols the composition
`
`according to the present invention contains
`
`kaempfero1-3-monoglucoside, quercetin-3—monog1ucoside.
`
`myricetin—3—monog1ucoside and astragalin. As sugars it
`
`contains D—g1ucose. D—fructose. saccharose, raffinose,
`
`arabinose. xylose. As free amino acids the composition
`
`according to the present invention contains lysine.
`
`histidine, arginine. aspartic acid,
`
`threonine, serine,
`
`glutamic acid. proline, glycine, alanine,
`
`
`
`
`
`12
`
`cystine, valine. methionine.
`
`isoleucine,
`
`leucine.
`
`tyrosine and phenylalanine. As organic acids it
`
`contains tartaric acid, malic acid, citric acid.
`
`5
`
`ascorbic acid. n-ketoglutaric acid,
`
`fumaric acid,
`
`LJI
`
`galacturonic acid. glyceric acid. glycolic acid,
`
`glycouronic acid. oxalic acid. succinic acid. shikimic
`
`acid. As sterols the composition according to the
`
`present
`
`invention contains 8—cetostero1. stigmasterol.
`
`kaempesterol.
`
`As methylsterols it contains
`
`10
`
`obtusifoliol. citrostadienol. As dimethylsterols it
`
`incorporates d—amyrin, B—amyrin.
`
`lnpeol,
`
`taraksterol,
`
`taraxasterol, germanicol. As
`
`lignans the
`
`composition according to the present invention contains
`
`oxymatairesinol. Natairesinol. pinoresionol.
`
`liovyl,
`
`13
`
`isolariciresinol and olivyl.
`
`As
`
`lignan glycosides it
`
`contains quetinol arabinoside and querinol xyloside.
`
`As
`
`phenolic acids it contains paraoxybenzoic acid.
`
`protocatechinic acid. gallic acid. vanillic acid and
`
`syringic phenolic acids.
`
`As phenolic aldehydes the
`
`20
`
`composition according to the present
`
`invention contains
`
`vanilline. syrinqic aldehyde. sinapic aldehyde and
`
`coniferyl aldehyde.
`
`As alkylferulates it contains alkyl
`
`esters of ferulic acid with the alcohol moiety being
`
`represented by octadecanol, eicosanol. docosanol.
`
`25
`
`tetracosanol. hexacosanol.
`
`The above—mentioned composition of
`
`the
`
`f:
`
`
`
`'-t.‘
`
`13
`
`herein—before—listed ingredients can also be obtained in
`
`the form of natural1y—occurring complexes of
`
`biologically active substances of
`
`the vegetable origin.
`
`The above-mentioned composition of the heroin-
`
`before—listed ingredients is soluble in water, ethanol
`
`and aqueous—alcoholic solutions.
`
`The composition according to the present
`
`invention
`
`has a low toxicity:
`
`LD50 is 36.5 ml per 1,000 g of
`
`body—mass of a rat.
`
`10
`
`15
`
`20
`
`We have carried out pharmacological studies of
`
`the
`
`effect of the composition according to the present
`
`invention on processes of ethanol consumption and on the
`
`formation of a physical dependence on animals and human
`
`beings.
`
`Under conditions of a chronic experiment
`
`(15 weeks)
`
`on mature male rats of Wistar line the level of ethanol
`
`consumption was studied under the conditions of free
`
`choice between water and a 15% ethanol. Prior thereto
`
`the rats were tested for resistance to ethanol by the
`
`"side posture" procedure upon an intraperitoneal
`
`H"-‘
`
`administration of
`
`a 25% ethanol at the rate of 4.5 g/kg
`
`of
`
`the body mass of
`
`the animals.
`
`In the experiment rats
`
`with similar characteristics of a high tolerance towards
`
`
`
`
`
`14
`
`ethanol were used. Later on the animal were placed into
`
`cases with calibrated drinking bowls under conditions of
`
`free choice between a 15% ethanol and water. and the
`
`daily consumption of
`
`the liquids was recorded.
`
`The control group was composed of animals (12 rats)
`
`that consumed a 15% ethanol.
`
`In the test group (12 rats) the composition
`
`according to the present invention was added to a 15%
`
`ethanol
`
`in the drinking bowl
`
`in the amount of 1 ml per
`
`10
`
`50 ml of 15% ethanol. After 13 weeks of active
`
`alcoholization the animals were deprived of
`
`the access
`
`to alcohol for 10 days (deprivation period) and then the
`
`amount of consumed solutions was recorded again.
`
`The
`
`experimental data are shown in Table 1.
`
`In the group of control animals the deprivation
`
`period proceeded with withdrawal phenomena which
`
`manifested in a changed behaviour of
`
`the animals, signs
`
`of
`
`tremor were recorded. a moderate dishevelling of hair
`
`was noted.
`
`At
`
`the same time.
`
`in the control group no
`
`20
`
`withdrawal symptoms were observed.
`
`The character of consumption of a 15% ethanol under
`
`free choice conditions in the control group was
`
`different from that of consumption of a 15% ethanol with
`
`‘I.
`
`
`
`
`
`1;‘.
`
`the composition according to the present invention in
`
`the test group. Beginning from the 3th weeks a clearly
`
`15
`
`pronounced trend towards reduction of ethanol
`
`consumption in combination with the composition
`
`according to the present invention was observed and
`
`after deprivation this difference was exceeding 100%.
`
`An important indicator of a formed physical dependence
`
`on ethano1_in the control group was an increased rate of
`
`ethanol consumption after a 10-days‘ deprivation by
`
`12%.
`
`In the test group the consumption of ethanol in
`
`combination with the composition according to the
`
`present invention after deprivation remained at the same
`
`level.
`
`TABLE 1
`
`Effect of the composition according to the present
`
`invention on the amount of consumed 15% ethanol on a
`
`daily basis (in ml/kg of 1 animal's bodyweight) under
`
`free choice conditions
`
`10
`
`15
`
`ill’
`
`
`
`
`
`Tnble 1
`
`rameter
`
`Time of consumption (in weeks)
`
`
`
`33.0
`3.55
`
`32-2
`4_og
`
`31.0
`2_g3
`
`91
`
`1
`9
`3
`11
`10
`9
`n
`7
`6
`5
`4
`15
`13
`12 ~
`1. Amount of
`20.9
`20.1
`22.5
`25.0 20.3 25.5 22.5 20.0 24.0 20.2 29.1
`29.4 24.1 Henri-'29.4
`consumed 15%
`1.06
`1.18
`2.29
`2.05 1.01.97 2.32 1.332.115 2.25 1.25
`3.44 1.29 "“E§°“ 3
`,2
`ethanol (con-
`days
`trul group)
`2. Amount of
`consumed 15%
`ethanol wlth
`addition of
`the compualv
`tion of this
`invention
`(1 ml per
`50 ml of et-
`hanol)
`
`34.0 25.9 26.6 29.3 15.3 10.9 22.0 22.0 20.0 13.0 E:f”ii3.0
`3_11
`2,5} 2.96 1.3
`1.49 1.16 2.1? 1.66 2.42 2.12 gig“
`2.41
`0
`
`0.0
`
`5
`
`0.0
`
`5
`
`0.01 0.2
`
`0.1
`
`-
`
`-
`
`0.01 0.001
`0_ou1
`
`0.2 0.1 0.05 1
`days U.0D
`
`time of consumptlcn (in weeks)
`
`
`
`17
`
`Addition,
`
`to a 15% ethanol. of
`
`the-composition
`
`according to the present invention under conditions of a
`
`lonq—time forced alcoholization (38 months) with the
`
`absence of water
`
`in the food diet has resulted in a
`
`substantial redistribution of animals in groups of
`
`alcohol consumption (Table 2).
`
`TABLE 2
`
`Effect of
`
`the composition according to the present
`
`invention on distribution of rats according to the rate
`
`10
`
`of consumption of a 15% ethanol (in per cent) {forced
`
`alcoholization).
`
`F-——__flH*___—_h__Fh—T__"—_"__—_—_”**'"‘”""1F‘”**”*"‘“
`fl
`_
`Alcoholization time
`
`uroups of animals
`
`8 months
`
`6 months
`
`3 months
`
`
`
`Low-drinking (20-60
`ml per 1,000 g of
`the bodymess)
`
`25/45
`
`73/76
`
`57/79
`
`Medium-drinking
`
`(so-so ml per 15000g I
`
`of the bodymsss
`
`31/12
`
`22/21
`
`21/15
`
`Heavily-drinking
`above 80 ml per 1,000g
`12/5
`5/3
`43/13
`of the bodymass)
`
`
`-1'}
`
`Q}
`
`*) Note:
`
`In the numerator — consumption of a 15%
`
`ethanol.
`
`in the denominator — consumption of a 15%
`
`ethanol with the addition of a composition according to
`
`the present
`
`invention.
`
`
`
`
`
`18
`
`The conditions of this experiment contemplated an
`
`individual control of consumption of test solutions in
`
`L!‘
`
`groups of animals:
`
`among the rats administered with
`
`alcohol
`
`incorporating the composition according to the
`
`present invention the number of heavily-drinking animals
`
`was certainly smaller.
`
`To avoid possible organoleptic effect of the
`
`composition according to the present
`
`invention on the
`
`level of ethanol consumption under free—choice
`
`10
`
`conditions parallel experiments have been carried out
`
`where the composition was introduced intragastrically.
`
`not into the test solution.
`
`The test results turned to
`
`be identical
`
`irrespective of
`
`the routes of
`
`administration of
`
`the composition according to the
`
`present
`
`invention.
`
`Gas-liquid chromatography was used to determine the
`
`amount of ethanol
`
`in blood of animals of
`
`the test and
`
`control groups that were given the test solution for the
`
`period of
`
`3 months.
`
`90 minutes prior to slaughtering
`
`20
`
`the animals they were introperitoneally administered
`
`with a 25% ethanol
`
`in combination with the composition
`
`according to the present invention in the ratio of 1:50
`
`{test group).
`
`Q:
`
`
`
`
`
`19
`
`The test results (Table 3) point to an essential
`
`increase (by more than 4 times) of ethanol in theblood
`
`of animals that were previously administered for a long
`
`time with the composition according to the present
`
`invention.
`
`The rate of elimination of ethanol from blood
`
`depend. first of all. on activity of alcoho1dehydro-
`
`genase (ADG) which has been studied against the
`
`background of an acute and chronical alcoholic
`
`10
`
`intoxication.
`
`Upon a sing1e—time intraperitoneal administration,
`
`to animals of a 15% ethanol
`
`in the does of 4.5 g/kg of
`
`the body mass, 30 minutes thereafter the activity of
`
`alcohol dehydrogenase is 3.51 mM/min/l relative to the
`
`intact group;
`
`the composition additive according to the
`
`present invention inhibits activity of enzymes in the
`
`presence of ethanol which is 5.86 mfi/min/1.
`
`In
`
`chronical experiments upon introduction of ethanol
`
`(passive alcoholization) over the period of 1.5 months
`
`20
`
`of a daily administration of a 15% ethanol and ethanol
`
`in combination with the composition according to the
`
`present invention in the test does of 1 g/kg the data
`
`have been obtained which prove the results of the
`
`previous experiment (see Table 4).
`
`H-9
`
`
`
`
`
`20
`
`Table 3
`
`Effect of
`
`the composition according to the present
`
`-2
`
`invention on elimination of ethanol after the addition
`
`of a 25: ethanol 4.5 g/kg of the animals‘ bodyweiqht-
`
`
`
`Experiment
`
`Statistical
`parameter
`
`Content of
`ethanol in
`blood, %
`
`
`
`1. Digested content (in-
`
`troduction of a 25%
`
`ethanol)
`
`7
`
`M;m
`
`O.72:0.14
`
`2. 3-months‘ consumption
`
`of a 15% ethanol (in-
`
`troduction of 25% et-
`hanol)
`14
`
`3. 3—nonths' consumption
`
`of e 15% ethanol in
`
`combination with the
`
`composition of the
`
`present invention
`
`Mim
`
`1.0:O.14
`
`(administration of 11
`
`25% ethanol)
`
`31$
`
`9
`
`2«5210-57
`
`0-01
`
`4. 3-months‘ consumption
`
`Of a 15% ethanol in
`
`combination with the
`
`composition of this
`
`invention (administ-
`ration of 25% ethe—
`nol + composition),
`
`1:50
`
`11
`
`firm
`P
`
`4-25:0-78
`0-001
`
`
`
`
`
`m
`
`21
`
`Table 4
`
`Activity of aicoholdehydzogenase in blood serum and
`
`liver upon administration of a 15% ethanol
`
`in
`
`combination with the composition according to the
`
`present invention orally for 1.5 months.
`
`A°t“dtY°f
`al°m“fl '
`denydrogenese
`in blood
`
`Activity of alcohol- Ethanol in
`dehydrogenase accord
`blood
`ing to Bonischoen
`me thod
`
`I
`
`serum by the
`Skursky method
`mnzmin/1
`
`1. 155 etLa-i
`n01»
`| 3-‘i1.17
`15% etha—\
`nol+com-;
`position 1
`
`2.
`
`_
`Lnrer.
`Serum
`mmrmin/1 mfi/min/1
`
`>1 ml
`
`3.15¢o.14
`
`47.02:1.91
`
`16.21i1.4
`
`2-7510.33
`
`2.21¢o.o5 34.39i2.6
`
`4.32¢o.4
`
`2-5110.29
`2-5:0.33
`
`2.51¢o.o6 4o.5i3.29
`2.46:0.09 4o.51¢z.3
`
`4.9¢o.6
`4.14:o.3
`
`II I II
`
`of this
`inventi-
`on
`
`3. Composi-
`tion of
`this in-
`vention
`(aqueous
`solution
`1:50)
`
`4. Physiolo-
`gical so-
`lution
`5- Intact
`
`I
`[2-63:0-49
`
`2.s6;o.05 37.4¢1.61
`
`21.s7¢2.5
`
`
`
`32
`
`Under conditions of free choice between a 15%
`
`ethanol and water (control group) and between a 15%
`
`ethanol with the composition according to the
`
`5
`
`invention and water after 1.5 and 3 months of
`
`U1
`
`consumption the activity of alcohol dehydrogenase was
`
`studied prior to and after deprivation.
`
`The results
`
`thus obtained are shown in Table 5.
`
`TABLE 5
`
`Activity of alcoholdehydrogenase at a free choice
`
`of
`
`the test solutions
`
`
`
`
`
`
`1.5 months of
`consumption
`
`3 months of
`consumption
`
`
` after
`prior to
`prior to
`oepriva—
`depri-
`depriva-
`
`
`tion
`tion
`vation
`
`
`
`1.
`
`¢-
`
`75% ethanol +
`
`15% ethanol
`
`2.7i0.26 3.61:O.48 2.64:0.27
`
`
`composition of
`i
`this invention 1.87iO.22|3.35¢0.44 3.95:0.66
`
`
`
`3- Intact
`4.33_+_1.o8 4.79_+o.64 2.2__-50.28
`3.08-1-0.58
`
`
`IL
`
`
`
`23
`
`Therefore.
`
`the composition additive according to the
`
`present invention decelerates oxidation of ethanol
`
`in
`
`the liver by inhibiting activity of alcohol
`
`dehydrogenase.
`
`Observations were carried out to study the lipoid
`
`and carbohydrate metabolism in animals upon
`
`administration of the composition according to the
`
`present
`
`invention against the background of a 3- and
`
`6-months' alcoholization.
`
`To this end. over the period
`
`10
`
`of
`
`3 and 6 months the rats were intragastrically
`
`administered with 2 ml of
`
`the hereinbe1ow—specified
`
`solutions per 100 q of the body mass.
`
`In the control:
`
`Group 1 distilled water: Group II — 15% ethanol: Group
`
`III — 15% ethanol containing a 5% composition according
`
`to the present invention; Group IV - aqueous solution
`
`of
`
`the composition according to the present invention.
`
`The results thus obtained are shown in Tables 6. 7,
`
`8 and 9 hereinbelow.
`
`1‘
`
`LE‘
`
`
`
`
`
`Variation of the content of neutral lipoidu in the liver of rats fed with the solutions
`for 3 months (in % of the total lipoidn, Qim)
`
`'i'.1|Il(- (1
`
`I
`
`C r o u p n
`II
`
`o f
`III
`
`a n i m a l s
`IV
`
`control
`
`distillate
`
`15% ethan01+
`aqueous solution
`of the comp0uiti— 0
`9-
`-0
`on of this inven-
`C mpo ltl n
`mm
`°f 32.11.33-
`
`V
`
`15% ethanol
`
`15,011.21
`12.510.48
`12.5iO.93
`14-510.49
`Cholesterol esters
`103
`B5
`86
`100
`% of variation
`20.1+1_O4 3)
`20.6+1.7T 3)
`20.1+0.84 3)
`1T.211.112)
`Triglycerides
`148
`151
`143
`127
`% of variation
`10.6_+_0.92 ”
`11510.53
`12.3¢0.50
`12,010.40
`Free fat acids
`_
`33
`90
`96
`100
`% of variation
`15.§11.22
`14.210.6T 1)
`16.1¢O.91
`17.21O.53
`Cholesterol
`92
`86
`97
`104
`% of variation
`Residual fraction
`32.5:1.05
`38.310.9T
`39.0;1.02
`41.211.30
`39.011.00
`
`
`:
`
`_
`
`?Z
`
`14,511.10
`
`13.E10.55
`
`1
`
`12.811.60
`
`16.6:O.9
`
`2) 1:40.02;
`1) P .:o.o5;
`p - probability
`
`3) 24.0.01
`
`
`
`'!'.-Ihll‘ ?
`
`Variation of Lhe content of neutral lipoido in the liver of rated feci with the
`em1u‘t:i.one: for 6 months (in ‘)5 of the total lipoids. 111m)
`
`Control
`
`I
`
`II
`
`Groups
`
`of
`
`Neutral Illpoidfi
`
`distillate
`
`15% ethanol
`
`cholesterol esters 16.91059
`K: of variation
`Triglycerides
`96 of variation
`Free fatty acids
`% of variation
`
`15.161021
`
`16.671o.4B
`
`16.‘3210.'?0
`99
`13.831312
`92
`1'r.21_.:0.11
`103
`
`_
`1E.E2_+_0.35
`100
`1a.19¢o.25‘)
`‘I20
`14.01049”
`84
`
`animals
`III
`
`1552’: ethaz_Iol+
`composition
`C’:
`thi’§' 1”‘
`‘''‘”“31°"
`
`16.T4;_l_0.27
`100
`14.1a¢o.22‘)
`94
`17.301037
`104
`
`IV
`
`aqueous fl01u_
`tion of the
`composition
`of the inven-
`tion
`15.81;Ir_0.‘N|
`95
`13.641042 1)
`90
`18.6610.88
`112
`
`if
`
`17.0610.19
`15.7210.39
`'|6.61i0.59
`17.0710.16
`Cholesterol
`99
`97
`96
`99
`% of variation
`Residual fraction
`333310.40
`353810.74
`34.58,1_0.47
`35.06_+_0.52
`343310.61
`
`
`17.283-_0.26
`
`1}
`2)
`
`P435053
`p<o.o2
`
`
`
`Variation of activity of lyaoaomal hydrulases in the liver of rats upon con—
`sumption of ethanol and the compoaition of this invention for 3 months
`and 6 months (nunomol/ml/min,m1mJ
`
`Groups of animals
`
`3 montha
`
`6
`
`months
`
`_fi— galactonidaac
`O.3310.0l
`03510.01
`106
`
`J6~g1uco0idnse
`D.4910.05
`093410.00
`110
`
`J9—ga1act00idnse
`0.3510.O1
`041310.02
`78
`
`J9— glycouidaue
`0.471G.G4
`0.43_1_0.01
`91
`
`0.58:0.08
`
`1)
`
`123
`
`0.50_t0.05
`106
`
`Control
`1. Distillate
`% of variation
`of the control
`II. 15 % ethanol
`% of variation
`of the control
`III. 15 95 ethanol + com-
`pofiiflon of this in_
`vention
`% of variation of
`the control
`IV. Aqueous solution of
`composition 01’ this
`invention
`% of variation of the
`control
`85
`94
`78
`70
`
`
`O.3710.04
`
`U.8710.09
`
`112
`
`178
`
`03510.06
`106
`
`O.6610.0EI
`135
`
`2)
`
`1)
`
`_
`
`O.8610.O6
`
`3)
`
`93
`
`247
`
`02610.01
`75
`
`0.25_1_0.01
`
`1)
`
`04010.02
`
`03110.03
`
`0.3.-3_+_0.05
`
`1} D¢:0.05;
`
`2) P:;0.01;
`
`3) D4:O.001
`
`1:
`
`
`
`£5‘
`
`'l'.'I 1: I v '1
`
`Variation of tho conteril. of carbohydra1.e-contaiuint; siinpuilymers. in the 1:'1.\rer of
`rats fed with ethanol and with. the con: ouition of the invention for 3 r:1ontl1::
`and 6 nronths
`in‘-_-_—%, Him)
`
`
`Groups of
`animals:
`
`
`llexoacs
`
`3 mon1;l1s;
`
`llexoaamines
`
`Iiuxoses
`
`6 months
`
`?
`liexonaniinesi
`
`1. control
`2. Distillate
`35 of variation
`of the control
`
`26.6£111.54
`1B.6T11.‘I2”
`
`70
`
`3. Aqueous solution
`of the composition
`of this invention
`3:3 of variation of
`the control
`
`1)
`
`333512-73
`125
`
`325312.31’
`29.c.u12.52-
`
`91
`
`36.02114?
`111
`
`26.:56—:1.«13
`20.45_-u_1.?2
`
`T8
`
`2B.9111.34
`110
`
`4. 15?; ethanol + c_:om—
`193111.10"
`234012.65
`ia.43;1.271)
`§f13:r*1fi';n°f this
`95 of variation
`75
`B6
`59
`of the control
`5.
`15% ethanol
`15.61+_1.22”
`253431.77“
`153211.002’
`9% of variation
`01‘
`the control
`62
`79
`E2
`
`
`1) P.-.—_:0.05;
`
`2)
`
`p-=:0.0l;
`
`3) p«-.:0.oo1
`
`£3
`
`4'3.0U11.76
`6.ri.53_+_6.972)
`
`140
`
`102.43;'r.633)
`209
`
`34.72+4.213’
`_
`173
`1)
`30.221541”
`62
`
`
`
`28
`
`Then we have carried out pharmacological tests of
`
`the composition according to the present invention as an
`
`agent for improving general resistance of
`
`the organism.
`
`For this purpose the effect of the composition according
`
`LJI
`
`to the present invention on the heat—resistance of rats
`
`has been studied.
`
`{1) Overheating of nondescript female rats (60
`
`animals)
`
`is effected by irradiation with an UHF—Eie1d by
`
`means of an instrument for a microwave therapy with the
`
`frequency of 2.375 mH2 — 17 mA for 10 days once a day
`
`over
`
`the period of 4 days.
`
`The test composition is
`
`administered in the dose of 2.5 ml/kg (intragastrically
`
`in all series of experiments) for 5 days before the
`
`beginning of
`
`irradiation and. on the say of experiment.
`
`one hour before irradiation.
`
`The death rate of rats is
`
`assessed after a 4~times'
`
`irradiation.
`
`It has been found that during one day after the last
`
`irradiation in the control group 22% of the animals
`
`died, whereas among the rats administered with the
`
`20
`
`composition according to the present
`
`invention the death
`
`rate was 11% {p< 0.01).
`
`(2) Overheating of male rats of
`
`the Wistar line is
`
`effected in a
`
`thermostatted cabinet at the temperature
`
`of 43°C.
`
`The test composition in the dose of 2.5 ml/kg
`
`
`
`
`
`29
`
`'
`
`(‘.1
`
`is administered for the preventive purposes over the
`
`period of 20 days.
`
`The rectal temperature and death
`
`rate of the animals are assessed.
`
`It has been found that in the control group 76% of
`
`5
`
`the animals (28 animals out of 37) died. while against
`
`the background of
`
`the composition according to the
`
`present invention 56% of
`
`the rats died (22 rats out of
`
`39: p< 0.001).
`
`The composition'provided no effect on
`
`the rectal temperature.
`
`10
`
`(3) Under
`
`the same conditions of overheating or male
`
`rats of the wistar line the composition according to the
`
`present invention is administered prophylactically over
`
`48 days in the dose of 1 ml/kg.
`
`It has been found that in the control group 54% of
`
`15
`
`rats (30 animals out of 55) died, while upon overheating
`
`against the background of a long—time administration of
`
`the composition according to the present invention the
`
`death rate was 42% (24 rats out of 57;
`
`p (0.001).
`
`(4) Overheating of male rats of the Wistar line was
`
`20
`
`effected in much the same manner.
`
`The test composition
`
`N
`
`was administered in the dose of 1 ml/kg 50 minutes prior
`
`to overheating.
`
`The overheating duration is 40 minutes
`
`
`
`30
`
`and 2 hours.
`
`The animals were killed by decapitation.
`
`Tested were:
`
`the content of glycogen {herein and in
`
`other cases — by the Zeifter method):
`
`activity of
`
`hexokinase and g1ucoso-6—phosphatedehydrogenase {herein
`
`and in other cases — by the formation of nicotinamide—
`
`dinuoleotidephosphoric acid (NADPH).
`
`It has been found that in overheating of
`
`the rats
`
`for 40 minutes the composition inhibited the drop of
`
`the
`
`content of glycogen, as well as of
`
`the activity of
`
`hexokinase and glucosewfi-phosphate dehydrogenase in the
`
`liver (see Table 10 hereinbelow).
`
`L_.rl
`
`10
`
`Upon overheating for 2 hours the test composition
`
`provided no effect on the level of
`
`the studied
`
`parameters.
`
`Overcooling of male rats of
`
`the Wistar line was
`
`caused by placing the animals into a refrigerator
`
`chamber at a
`
`temperature of 5°C for 1 and 2 hours.
`
`The
`
`compos