SE179098C1

SE179098C1 -

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Publication number: SE179098C1
Authority: SE
Publication date: 1962-01-01

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Uppfinnare: C W F Damaskus Prioritet begard fain den 3 juni 1953 (USA) Foreliggande uppfinning avser ett satt att framstalla ett preparat, som innehaller adrenokortikotropt hormon, vilket preparat lam-par sig for intramuskular eller subkutan injektion och uppvisar okad adrenokortikotrop verkan. Inventor: C W F Damascus Priority Required June 3, 1953 (USA) The present invention relates to a method of preparing a composition containing adrenocorticotropic hormone, which composition lambs for intramuscular or subcutaneous injection and exhibits increased adrenocorticotropic activity.

Adrenokortikotropt hormon Sr ett amne, som utsondras fran hypofys ens framlob och antagligen av blodstrommen i kroppen fores till binjuren, dar det utovar en inverkan pa utvecklingen, tillvaxten och aktiviteten av binjurebarken. Under senare Sr ha extrakt av adrenokortikotropt hormon, utvunnet ur dj urhypofyser, t. ex. fran svin, far ()eh notkreatur, visat sig utomordentligt verksamma vid be-handling av vissa sjuldiga tillstand has manniskor, i det att hormonet injiceras parenteralt, och det har uppkommit en standigt stigande efterfragan efter preparat av detta hormon lampade for injektion i manniskokropp en. Adrenocorticotropic hormone is a substance that is secreted from the pituitary gland and probably carried by the bloodstream in the body to the adrenal glands, where it has an effect on the development, growth and activity of the adrenal cortex. In later Sr have extracts of adrenocorticotropic hormone, extracted from dj ur pituitaries, e.g. from pigs, far () eh cattle, has been shown to be extremely effective in the treatment of certain rare human beings, in that the hormone is injected parenterally, and there has been a steadily increasing demand for preparations of this hormone lamped for injection into human bodies. .

Med hansyn till de f5rhallandevis knappa mangder, van i sadana djurhypofyser aro tillgangliga, ur vilka adrenokortikotropt hormon utvinnes, och Sven med hansyn till dessa hypofysers utomordentligt ringa storlek och ringa halt av verksamt amne Sr det av stor betydelse att ha tillgang till metoder icke blott for ernaende av storsta mojliga utbyte av aktivt amne ur de forhandenvarande kortlarna utan Sven for Mining av effekten hos det aktiva amnet i sh hog grad som mojligt. In view of the relatively scarce amounts accustomed to such animal pituitaries are available, from which adrenocorticotropic hormone is extracted, and Sven in view of the extremely small size and small content of active substance in these pituitary glands, it is of great importance to have access to methods not only for ernaende of the largest possible exchange of active subject from the available cards without Sven for Mining of the effect of the active subject to as high a degree as possible.

Enligt en hand metod har det visat sig, att det adrenokortikotropa hormonet, da det anvandes i forbindelse med gelatin som barsubstans' Sr betydligt mera verhsamt an samma mangd adrenokortikotropt hormon, som injiceras i form air en vanlig vattenlosning. SSsom namnt i forbindelse med denna kanda metod innebar Okningen av den effekt, som erMlles vid anvandande av gelatin, en fordubbling eller tredubbling av den andrenokortiko tropa verkan. Det har visat sig, att gelatinet medfor den verkan pa det adrenokortikotropa hormonet, att man genast uppnar en okning av den hormonala effekten, och att den fysiologiska verkningsbilden Sr i huvudsak densamma som den vilken orsakas av en vattenlosning av hormonet bortsett fran att blott en mycket mindre dos kraves, da man anvander en kombination av det adrenokortikotropa hormonet och gelatin, an nar man anvander en vattenlosning av hormonet. Denna mycket speciella verkan av gelatin pa adrenokortikotropt hormon har icke pavisats for de andra barsubstanser, som i allmanhet anvandas i injicerbara preparat. According to one method, it has been found that the adrenocorticotropic hormone, when used in conjunction with gelatin as a carrier substance, is significantly more potent than the same amount of adrenocorticotropic hormone injected into the air as a normal aqueous solution. As mentioned in connection with this known method, the increase in the effect obtained with the use of gelatin meant a doubling or tripling of the andrenocortico - tropical effect. It has been found that the gelatin has the effect of the adrenocorticotropic hormone, that one immediately achieves an increase in the hormonal effect, and that the physiological action picture is essentially the same as that which is caused by an aqueous solution of the hormone except that only a very a smaller dose is required when using a combination of the adrenocorticotropic hormone and gelatin than when using an aqueous solution of the hormone. This very special effect of gelatin on adrenocorticotropic hormone has not been demonstrated for the other carriers that are commonly used in injectable preparations.

Det Sr ett andamal med foreliggande uppfinning att astadkomma en metod for fram- tallning av ett preparat innehallande adrenokortikotropt hormon, som uppvisar en annu kraftigare verkan an den som Sr kand fran den ovannamnda metoden. Ett annat andamal Sr att astadkomma en metod fdr framstall, ning av en kombinationsprodukt av adrenokortikotropt hormon och en speciell gelatinbarsubstans, som Sr oshadlig och lampar sig for intramuskular eller subkutan injektion, varjamte den samtidigt astadkommer en samverkan mellan hormonet och barsubstansen, som medf5r en vasentligt storre okning av den adrenokortikotropa verkan an den som ernas med andra typer ay gelatin. It is an object of the present invention to provide a method for the development of a preparation containing adrenocorticotropic hormone, which has an even stronger effect than that known from the above-mentioned method. Another purpose is to provide a method for the preparation of a combination product of adrenocorticotropic hormone and a special gelatin bar substance, which is harmless and illuminates for intramuscular or subcutaneous injection, while at the same time providing a synergistic interaction between the hormone and the bar substance. greater increase in the adrenocorticotropic effect than that of other types of gelatin.

Foreliggande metod for framstallning av ett preparat, som innehaller adrenokortikotropt hormon, kannetecknas darav, att det adrenokortikotropa hormonet forsattes med oxipolygelatin. The present method for preparing a preparation containing adrenocorticotropic hormone may be characterized in that the adrenocorticotropic hormone is continued with oxypolygelatin.

Den produkt, som framstalles enligt uppfinningen, Sr en produkt innehallande det adrenokortikotropa hormonet och oxipolygelatin. Det adrenokortikotropa hormonet anvant i kombination med oxipolygelatin sasom barsubstans har en vasentligt storre effekt Sn 2— — samma mangd adrenokortikotropt hormon injicerat tillsammans med vilken som heist annan kand typ av gelatin och är naturligtvis annu kraftigare verkande an samma hormon I vanlig vattenlasning. Den &fling av verkan, som eras vid anvandande av oxipolygelatin, har visat sig innebara en fordubbling eller tredubbling av den adrenokortikotropa effekten matt i forhallande till den, som uppnas med andra typer av gelatin. The product prepared according to the invention is a product containing the adrenocorticotropic hormone and oxypolygelatin. The adrenocorticotropic hormone used in combination with oxypolygelatin as a bar substance has a significantly greater effect Sn 2 - the same amount of adrenocorticotropic hormone injected together with any other known type of gelatin and is of course even more potent than the same hormone in ordinary water reading. The effect of the use of oxypolygelatin has been found to be a doubling or tripling of the adrenocorticotropic effect in relation to that obtained with other types of gelatin.

Den ovan beskrivna almingen av effekten innebar en verklig Caning till skillnad fran en endast sadan forlanging av effekten som ibland kan orsakas av barsubstansens viskositet. Detta framgar av det faktum, att forbindelsen mellan oxipolygelatin och hormon omedelbart efter injektion astadkommer en plots-hg stark verkan, som vida overstiger den, vilken orsakas av en lika stor dos av hormonet I en vattenlosning, i vilken oxipolygelatin lace fir tillstades. Det her visat sig, att under dygnet efter en injektion den fysiologiska verkningsbilden far en given dos av oxipolygelatin-adrenokortikotropt hormon liknar den for en identisk dos av en saltlosning av adrenokortikotropt hormon bortsett frau att verkan av oxipolygelatin-adrenokortikotropt hormon vid en godtycklig tidpunkt ar storre, sasom om man hade injicerat en mycket stone dos. Denna verkan kan joke forklaras blott pa basis av en absorption, medforande en forlangd verkan, utan maste anses innebara en hell speeiell egenskap hos oxipolygelatin, som upptrader, dá detta amne anvandes tillsammans med adrenokortikotropt hormon, men som icke visar sig, da det anvandes i forening med andra biologiskt aktiva amnen. The generalization of the effect described above meant a real caning, in contrast to only such a demand of the effect which can sometimes be caused by the viscosity of the bar substance. This is evident from the fact that the connection between oxypolygelatin and hormone immediately after injection produces a sudden strong effect which far exceeds that caused by an equal dose of the hormone in an aqueous solution in which oxipolygelatin lace fir was allowed. It has been found that during the day following an injection the physiological action picture receives a given dose of oxipolygelatin-adrenocorticotropic hormone similar to that of an identical dose of a saline solution of adrenocorticotropic hormone except that the action of oxipolygelatin-adrenocorticotropic hormone at any time , as if a very stone dose had been injected. This effect can be joked explained only on the basis of an absorption, giving rise to a desired effect, but must be considered to have a very special property of oxypolygelatin, which occurs when this substance is used together with adrenocorticotropic hormone, but which does not show up when used in association with other biologically active substances.

Oxipolygelatin, den barsubstans som anvandes enligt foreliggande uppfinning, as en val kand produkt, som Or beskriven i litteraturen, och som kan erhallas sasom vanlig handelsprodukt (se t. ex. en artikel av Campbell et al med titeln The Preparation and Properties of a Modified Gelatin (Oxipolygelatin) as an Oncotic Substitute for Serum Albumin», Texas Reports on Biology and Medicine, bd 9, nr 2, sid. 235-280, sommaren 1951). Van_ ligtvis framstalles oxipolygelatin genom koppling av gelatinmolekylerna till starre komplex med ett kopplande amne, varefter den hogmolekylara gelatinpolymeren underkastas oxidation, varigenom molekylen nedbrytes till anskad storlek och konfiguration. En lamplig metod for framstallning av oxipolygelatin innefattar kondensation av gelatin med glyoxal och efterfoljande oxidation med vateperoxid. Det antages, att det vid reaktionen sker en andring av det ursprungliga gelatinets tenforformiga molekyl till en molekyl, som Or mera kulformig. Oxypolygelatin, the bar substance used according to the present invention, is a well-known product, as described in the literature, and which can be obtained as a common commercial product (see, e.g., an article by Campbell et al entitled The Preparation and Properties of a Modified Gelatin (Oxipolygelatin) as an Oncotic Substitute for Serum Albumin », Texas Reports on Biology and Medicine, vol. 9, no. 2, pp. 235-280, summer 1951). Usually, oxypolygelatin is prepared by coupling the gelatin molecules to rigid complexes with a coupling agent, after which the high molecular weight gelatin polymer is subjected to oxidation, whereby the molecule is degraded to the desired size and configuration. A suitable method for producing oxypolygelatin involves condensation of gelatin with glyoxal and subsequent oxidation with hydrogen peroxide. It is believed that in the reaction, the tin-shaped molecule of the original gelatin changes to a molecule which is more spherical.

Oxipolygelatin har framstallts och anvandes huvudsakligen sfisom plasmautspadningsmedel, och dess speciella verkan, dO det anvandes sasom barsubstans for adrenokortiko tropt hormon, har hittills kite varit kand. man emellertid tager 1 betraktande dess kanda egenskaper, nanaligen att det Or ogiftigt, icke antigent och icke innehaller pyrogena amnen avensom den hittills okanda egenskapen, att det Okar verkan av adrenokortikotropt hormon, sO. inses, att anvandbarheten sasom barsubstans for detta hormon Or en utomordentligt viktig och nyttig egenskap. Oxypolygelatin has been produced and used mainly as a plasma diluent, and its special effect, when used as a carrier substance for adrenocorticotropic hormone, has so far been known. however, one takes into account its known properties, namely that it is non-toxic, non-antigenic and does not contain pyrogenic substances in addition to the hitherto unknown property, that it increases the effect of adrenocorticotropic hormone, sO. It will be appreciated that the utility as a bar substance for this hormone is an extremely important and useful property.

Vid framstallning av produkten enligt uppfinningen Or det att foredraga att anvanda en vattenlosning av oxipolygelatin med en halt av 35-%. Det Or vanligtvis anskvart, att koncentrationen av oxipolygelatin ligger ffiom omradet c:a 5 till e:a 50 vikt% av den losning, som skall anvandas for injektion, Oven om oxipolygelatinets formaga att Oka det adrenokortikotropa hormonets effekt till en viss grad ocksa gar sig gallande, om koneentrationen av oxipolygelatin ligger Over eller under dessa granser. When preparing the product according to the invention, it is preferable to use an aqueous solution of oxypolygelatin with a content of 35%. It is usually assumed that the concentration of oxypolygelatin is in the range of about 5 to about 50% by weight of the solution to be used for injection. Although the ability of oxypolygelatin to increase the effect of the adrenocorticotropic hormone to some extent also increases, bile, if the concentration of oxypolygelatin is Above or below these limits.

Vilken som helst form av det adrenokortikotropa hormonet kan anvandas tillsammans med oxipolygelatin f8r att astadkomma den farbattrade adrenokortikotropa verkan, som innefattas av uppfinningen. Det adrenokortikotropa hormonet bur annu icke isolerats och dess molekylara struktur har annu icke beskrivits pa belt bestamt satt. Man anser, att hormonet Or ett figgviteamne, och vissa processer vid framstallningen anses medfara att en polypeptidmolekyl bildas, som utovar den adrenokortikotropa verkan. Enligt uppfinningen foredrages anvandningen ay ett golf, rent preparat, men ett godtyckligt ay de Ian& adrenokortikotropa hormonpreparaten, antingen de besta av det s. k. »hela aggviteamnet» eller ay »polypeptiden» och oberoende ur vilket ramaterial de utvunnits och vilken framstallningsmetod som kommit till anvand- kan anvandas tillsammans med oxipolygelatin fOr uppnaende av de forbattrade resultaten enligt uppfinningen. Any form of the adrenocorticotropic hormone may be used in conjunction with oxypolygelatin to provide the enhanced adrenocorticotropic action contained in the invention. The adrenocorticotropic hormone should not yet be isolated and its molecular structure has not yet been described in a definite way. The hormone Or is believed to be a fig white matter, and certain processes in the preparation are thought to result in the formation of a polypeptide molecule which exerts the adrenocorticotropic action. According to the invention, the use is preferred in a golf, pure preparation, but an arbitrary use in the Ian & adrenocorticotropic hormone preparations, either the best of the so-called "whole agglomerate" or the "polypeptide" and regardless of the raw material from which they are extracted and the method of preparation used. can be used together with oxypolygelatin to achieve the improved results of the invention.

Det sail, varpa det aktiva amnet och oxipolygelatin samverka f8r astadkommande av den forbattrade verkan Or f. n. icke med sakerhet kfint. En majlig forklaring kan vara, att den oxidationsreaktion, vilken oxipolygelatin underkastas, tillfor molekylen ett okat antal sura karboxylgrupper, som kunna fistedkomma att oxipolygelatinet bindes vid det basiska adrenokortikotropa hormonet, sa att det bildas ett kemiskt komplex med Rad adrenokortikotrop verkan. En ytterligare eller kanske annan forklaring Or att oxipolygelatin, nar det anvandes i samband med det adrenokortikotropa hormonet, kan forete en specifik, stabiliserande verkan p0 hormonet, sO att det skyddar detta mot inaktivering genom vissa enzymer eller andra forstorande krafter, som verka vid injektionsstallet. Under alla omstandigheter visa bade effektbestamningar och kliniska undersokningar pa patienter, som behandlats med adrenokorti- — —3 kotropt hormon, att anvandningen av oxipolygelatin medfor en vasentligt kraftigare verkan fran varje dos av hormonet an fallet är, da man anvander en saltlosning eller andra typer av gelatin som barsubstans. The sail, warp the active substance and oxypolygelatin interact to achieve the improved effect Or f. N. Not with certainty kfint. One possible explanation may be that the oxidation reaction to which oxypolygelatin is subjected gives the molecule an increased number of acidic carboxyl groups, which may cause the oxypolygelatin to bind to the basic adrenocorticotropic hormone, so that a chemical complex with Rad adrenocorticotropic action is formed. A further or perhaps different explanation is that oxypolygelatin, when used in conjunction with the adrenocorticotropic hormone, may have a specific, stabilizing effect on the hormone, so that it protects it from inactivation by certain enzymes or other magnifying forces acting at the injection site. In any case, both efficacy assessments and clinical trials in patients treated with adrenocorticotropic hormone show that the use of oxypolygelatin has a significantly stronger effect on each dose of the hormone than when using a saline solution or other types of gelatin as a bar substance.

Uppfinningen kommer i det foljande att forklaras i samband med exempel. The invention will be explained in the following in connection with examples.

Exempel 1. En portion av det adrenokortikotropa hormonet framstalldes genom extraktion med isattika vid 70° C av svinhypofysframlober, som avvattnats med aceton. Darefter tillsattes en halv volym aceton och en lietn portion natriumklorid till losningen for att utfalla icke onskade amnen. En liten volym etyleter tillsattes till losningen for att utfalla en raprodukt av det adrenokortikotropa hormonet. Fallningen renades genom adsorption pa oxicellulosa och eluering harifran, och ett jonbytande harts tills attes eluatet for att omvandla hormonet till acetatformen. Den pa detta salt framstallda losningen frystorkades, sit att ett torrt pulver bildades, vilket upplostes i pyrogenfritt, destillerat vatten. L8sningen sterilfiltrerades och infordes i glas med 40 enheter per glas, varefter innehallet frystes och frystorkades. Example 1. A portion of the adrenocorticotropic hormone was prepared by extraction with glacial acetic acid at 70 ° C from porcine pituitary lobes dehydrated with acetone. Then half a volume of acetone and a small portion of sodium chloride were added to the solution to precipitate unwanted substances. A small volume of ethyl ether was added to the solution to precipitate a rap product of the adrenocorticotropic hormone. The precipitate was purified by adsorption on oxicellulose and eluting therefrom, and an ion exchange resin until the eluate was added to convert the hormone to the acetate form. The solution prepared on this salt was lyophilized until a dry powder formed, which was dissolved in pyrogen-free distilled water. The solution was sterile filtered and introduced into glasses at 40 units per glass, after which the contents were frozen and lyophilized.

Den ovannamnda frystorkade produkten, vilken ar av den typ, som stundom kallas polypeptidformen av det adrenokortikotropa hormonet, uppl8stes i en losning innehallande 37 % oxipolygelatin och 0,5 % fenol. Den erhallna losningen, som inneholl det adrenokortikotropa hormonet, 37 % oxipolygelatin och 0,5 % fenol, underkastades de vanliga kliniska proven och visade sig ha 2-3 ganger sit kraftig verkan som det vanliga adrenokortikotropa hormonet givet tillsammans med vanlig gelatin. The above lyophilized product, which is of the type sometimes called the polypeptide form of the adrenocorticotropic hormone, was dissolved in a solution containing 37% oxypolygelatin and 0.5% phenol. The resulting solution, which contained the adrenocorticotropic hormone, 37% oxypolygelatin and 0.5% phenol, was subjected to the usual clinical trials and was found to have 2-3 times its potent effect as the usual adrenocorticotropic hormone given together with regular gelatin.

Den ovannamnda losningen av oxipolygelatin framstalldes av en vanlig 18sning av oxipolygelatin i form av en handelsprodukt. Denna vanliga losning framstalldes genom kondensation av gelatin med glyoxal foljt av oxidation med vateperoxid, och handelsprodukten astadkoms i form av en 5 %-ig losning av oxipolygelatin i normal saltlosning. Denna produkt bereddes for anvandning i föreliggande exempel pa sit satt, att saltet avlagsnades genom dialys, varefter 18sningen koncentrerades i vakuum genom destillation tills koncentrationen 'Mats till 37 % oxipolygelatin, varefter 0,5 % fenol tillsattes och det hela sterilfiltrerades. Den harigenom erhallna losningen anvandes sedan tillsammans med det frystorkade adrenokortikotrOpa hormonet sasom ovan beskrivits. The above-mentioned solution of oxypolygelatin was prepared from a common solution of oxipolygelatin in the form of a commercial product. This usual solution was prepared by condensation of gelatin with glyoxal followed by oxidation with hydrogen peroxide, and the commercial product was obtained in the form of a 5% solution of oxypolygelatin in normal saline. This product was prepared for use in the present example in such a way that the salt was removed by dialysis, after which the solution was concentrated in vacuo by distillation until the concentration was adjusted to 37% oxypolygelatin, after which 0.5% phenol was added and the whole was sterile filtered. The resulting solution was then used in conjunction with the lyophilized adrenocorticotropic hormone as described above.

Exempel 2. Frystorkat adrenokortikotropt hormon, framstallt sasom exempel 1, upplostes I en. 16 %-ig gelatinlosning, sit att en produkt med adrenokortikotrop verkan pa 16,5 enheter per milliliter erholls. Example 2. Freeze-dried adrenocorticotropic hormone, prepared as in Example 1, was dissolved in a. 16% gelatin solution, provided that a product with an adrenocorticotropic effect of 16.5 units per milliliter is obtained.

Denna losning delades i tva lika stora delar och anvandes for framstallning av de ne dan namnda losningarna A och B pa foljande satt: Lasning A framstalldes genom utspadning av utgangslosningen till 0,010 enheter per 0,5 ml med 16 %-ig gelatinlosning innehallande 0,5 % fenol. This solution was divided into two equal parts and used to prepare the following solutions A and B in the following manner: Solution A was prepared by diluting the starting solution to 0.010 units per 0.5 ml with 16% gelatin solution containing 0.5 % phenol.

Losning B framstalldes genom utspadning av utgangslosningen till 0,040 enheter per 0,5 ml med en 37 %-ig losning av oxipolygelatin innehallande 0,% fenol. Solution B was prepared by diluting the starting solution to 0.040 units per 0.5 ml with a 37% solution of oxypolygelatin containing 0.1% phenol.

Losningarna A och B provades darefter genom standardmetoden, varvid den adrenala utskiljningen av askorbinsyran provades pa hypofysektomiserade rattor. Det visade sig, att losning B, som innehaller oxipolygelatin, hade en verkan, som var c:a 150-200 % storre an verkan av losning A, vilken endast inneholl vanligt gelatin. Solutions A and B were then tested by the standard method, the adrenal excretion of ascorbic acid being tested on pituitary-stained dials. It was found that solution B, which contained oxypolygelatin, had an effect which was about 150-200% greater than the effect of solution A, which contained only ordinary gelatin.

Exempel 3. Frystorkat adrenokortikotropt hormon, framstallt pa samma salt som i exempel 1, upplostes i 16 %-ig gelatin, sit att en produkt med en verkan av 16,5 enheter per 0,5 ml erholls. Example 3. Freeze-dried adrenocorticotropic hormone, prepared on the same salt as in Example 1, was dissolved in 16% gelatin, yielding a product having an effect of 16.5 units per 0.5 ml.

Denna 15sning delades i tva lika stora delar och anvandes till framstallning av de nedan omtalade 18sningarna C och D pa foljande satt: Losning C framstalldes genom utspadning av utgangslosningen till 0,120 enheter per 0,5 ml med en 16 %-ig losning av gelatin innehallande 0,5 % fenol. This solution was divided into two equal parts and used to prepare the solutions C and D mentioned below as follows: Solution C was prepared by diluting the starting solution to 0.120 units per 0.5 ml with a 16% gelatin solution containing 0 , 5% phenol.

Losning D framstalldes genom utspadning av utgangslosningen till 0,120 enheter per 0,5 ml med en 37 %-ig losning av oxipolygelatin innehallande 0,5 % fermi. Solution D was prepared by diluting the starting solution to 0.120 units per 0.5 ml with a 37% solution of oxypolygelatin containing 0.5% fermi.

Losning C och D provades genom standardmetoden, varvid den adrenala utskiljningen av askorbinsyra provades pa hyofysektomiserade rattor. Det visade sig, att losning D, vilken inneholl oxipolygelatin, hade en verkan som uppgiek till 150-200 % av verkan has losningen C, vilken endast inneholl vanligt gelatin. Solutions C and D were tested by the standard method, the adrenal excretion of ascorbic acid being tested on hyophysectomized rats. It was found that solution D, which contained oxypolygelatin, had an effect equal to 150-200% of the effect of solution C, which contained only ordinary gelatin.

De ovannamnda enheterna i samband med verkan av det adrenokortikotropa hormonet aro den internationella enhet, som godkants av World Health Organization. En internationell enhet definieras som svarande mot verkan av ett milligram av preparatet LA-1-A, dk detta provas i overensstammelse med den metod, som beskrivits av M. Sayers, G. Sayers oeh L. A. Woodbury i tidskriften Endocrinology, 42, 379 (1948). The above-mentioned units in connection with the action of the adrenocorticotropic hormone are the international unit approved by the World Health Organization. An international unit is defined as corresponding to the action of one milligram of the preparation LA-1-A, dk this is tested in accordance with the method described by M. Sayers, G. Sayers and LA Woodbury in the journal Endocrinology, 42, 379 (1948 ).

Far att visa effekten av olika oxipolygelatinkoncentrationer jamforda med ACTH-gelatin-standardpreparatet innehallande 16 % gelatin hanvisas till foljande forsoksresultat. Forsoken avse sterogen effekt hos marsvin. Varje varde avser den steroida utsondringen hos en grupp om 8 djur. 1/3 av totala dosen gays intramuskulart med 12 timmars mellanrum; 3 ganger under 48 timmars urinuppsamling, vars 17-hydroxikortikoidhalt jamfordes med den under en 48 timmars kontrollperiod. To show the effect of different oxypolygelatin concentrations compared to the standard ACTH gelatin preparation containing 16% gelatin, reference is made to the following test results. The experiments concerned the sterogenic effect in guinea pigs. Each value refers to the steroid secretion in a group of 8 animals. 1/3 of the total dose is given intramuscularly at 12-hour intervals; 3 times during 48 hours of urine collection, the 17-hydroxycorticoid content of which is compared with that of a 48 hour control period.

— — Resultatet yid injicering ay ett ACTH-preparat innehallande 16 % yanIigt gelatin: En dos om 2,0 U.S.P.-enheter gay 200 % inning ay utsondringen ay 17-hydroxikortikoid. - - The result of injection of an ACTH preparation containing 16% yanIlig gelatin: A dose of 2.0 U.S.P. units gay 200% inhalation of the secretion of 17-hydroxycorticoid.

En dos om 1,0 U.S.P.-enheter gay 130 % okning av utsondringen av 17-hydroxikortikoid. A dose of 1.0 U.S.P. units gay 130% increase in the secretion of 17-hydroxycorticoid.

En dos om 0,5 g U.S.P.-enheter gay 70 % okning av utsondringen av 17-hydroxikortikoid. A dose of 0.5 g U.S.P. units gay 70% increase in the secretion of 17-hydroxycorticoid.

Da ett ACTEI-preparat innehallande 16 % oxipolygelatin injicerades erhtillos foljande resultat: En dos om 1,0 U.S.P.-enheter gay 160 % okning ay utsondringen ay 17-hydroxikortikoid. When an ACTEI preparation containing 16% oxypolygelatin was injected, the following results were obtained: A dose of 1.0 U.S.P. units gay 160% increase in the secretion of 17-hydroxycorticoid.

En dos om 0,5 U.S.P.-enheter gay 1% Mining ay utsondringen ay 17-hydroxikortikoid. A dose of 0.5 U.S.P. units gay 1% Mining ay excretion ay 17-hydroxycorticoid.

Da ett ACTH-preparat innehallande 20 % oxipolygelatin injicerades erhollos foljande resultat: En dos om 1,0 U.S.P.-enheter gay 190 % Mining av utsondringen av 17-hydroxikortikoid. When an ACTH preparation containing 20% oxypolygelatin was injected, the following results were obtained: A dose of 1.0 U.S.P. units gay 190% Mining of the secretion of 17-hydroxycorticoid.

Da ett ACTH-preparat innehallande 28 % oxipolygelatin injicerades erhollos foljande resultat: En dos om 1,0 U.S.P.-enheter gay 210 % Caning ay utstindringen ay 17-hydroxikortikoid. When an ACTH preparation containing 28% oxypolygelatin was injected, the following results were obtained: A dose of 1.0 U.S.P. units gay 210% Caning ay the excretion ay 17-hydroxycorticoid.

En dos om 0,5 U.S.P.-enheter gay 150 % Wining av utsondringen av 17-11.3-droxikortikoid. A dose of 0.5 U.S.P. units gay 150% Wining of the secretion of 17-11.3-droxycorticoid.

Dâ ett ACTH-preparat innehallande 37 % oxipolygelatin injicerades erhollos f8ljande resultat: En dos om 1,0 U.S.P.-enheter gay 270 % Wining ay uts5ndringen ay 17-hydroxikortikoid. When an ACTH preparation containing 37% oxypolygelatin was injected, the following results were obtained: A dose of 1.0 U.S.P. units gay 270% Wining ay the excretion of 17-hydroxycorticoid.

En dos om 0,67 U.S.P.-enheter gay 195 % alining ay utsondringen ay 17-hydroxikortikoid. A dose of 0.67 U.S.P. units gay 195% alining ay excretion ay 17-hydroxycorticoid.

En dos mil 0,33 U.S.P.-enheter gay 160 % Wining ay utsondringen ay 17-hydroxikortikoid. One dose mile 0.33 U.S.P. units gay 160% Wining ay excretion ay 17-hydroxycorticoid.

En. dos om 0,17 U.S.P.-enheter gay 57 % okning ay utsondringen ay 17-hydroxikortikoid. One. dose of 0.17 U.S.P. units gay 57% increase in secretion ay 17-hydroxycorticoid.

Claims

Patent claim:

1. Prepared to produce a preparation containing adrenocorticotropic hormone, characterized in that the adrenocorticotropic hormone was continued with oxypolygelatin.

2. According to claim 1, it is claimed that an aqueous solution with 35-40% of oxypolygelatin was used.

Salt according to claims 1 and 2, characterized in that oxypolygelatin was added in the side amount, that the concentration of the finished preparation amounts to 5%. Request publications: U.S. Patent Nos. 2,591,133, 2,669,537. Stockholm 1962. liungl. Boktr. P. A. Norstedt & Boner. 626080