DK149134B - METHOD FOR PREPARING THE DINATRUM SALT OF N- (PHOSPHONOACETYL) -L-ASPARAGINIC ACID - Google Patents

METHOD FOR PREPARING THE DINATRUM SALT OF N- (PHOSPHONOACETYL) -L-ASPARAGINIC ACID Download PDF

Info

Publication number
DK149134B
DK149134B DK506278AA DK506278A DK149134B DK 149134 B DK149134 B DK 149134B DK 506278A A DK506278A A DK 506278AA DK 506278 A DK506278 A DK 506278A DK 149134 B DK149134 B DK 149134B
Authority
DK
Denmark
Prior art keywords
pala
liters
salt
phosphonoacetyl
water
Prior art date
Application number
DK506278AA
Other languages
Danish (da)
Other versions
DK506278A (en
DK149134C (en
Inventor
Robert John Schultz
Fred William Starks
Original Assignee
Starks Ass
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Starks Ass filed Critical Starks Ass
Publication of DK506278A publication Critical patent/DK506278A/en
Publication of DK149134B publication Critical patent/DK149134B/en
Application granted granted Critical
Publication of DK149134C publication Critical patent/DK149134C/en

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
    • C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic Table
    • C07F9/02—Phosphorus compounds
    • C07F9/28—Phosphorus compounds with one or more P—C bonds
    • C07F9/38—Phosphonic acids [RP(=O)(OH)2]; Thiophosphonic acids ; [RP(=X1)(X2H)2(X1, X2 are each independently O, S or Se)]
    • C07F9/3804—Phosphonic acids [RP(=O)(OH)2]; Thiophosphonic acids ; [RP(=X1)(X2H)2(X1, X2 are each independently O, S or Se)] not used, see subgroups
    • C07F9/3808—Acyclic saturated acids which can have further substituents on alkyl
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/66—Phosphorus compounds
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
    • C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic Table
    • C07F9/02—Phosphorus compounds
    • C07F9/28—Phosphorus compounds with one or more P—C bonds
    • C07F9/38—Phosphonic acids [RP(=O)(OH)2]; Thiophosphonic acids ; [RP(=X1)(X2H)2(X1, X2 are each independently O, S or Se)]
    • C07F9/40—Esters thereof
    • C07F9/4003—Esters thereof the acid moiety containing a substituent or a structure which is considered as characteristic
    • C07F9/4006—Esters of acyclic acids which can have further substituents on alkyl

Landscapes

  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Animal Behavior & Ethology (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Biochemistry (AREA)
  • Molecular Biology (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • General Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Epidemiology (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Description

149134149134

Opfindelsen angår en fremgangsmåde til fremstilling af det hidtil ukendte dinatriumsalt af N-(phospho-noacetyl)-L-asparaginsyre. Den frie tetravalente syre N-(phosphonoacetyl)-L-asparaginsyre (i det følgende i 5 visse tilfælde betegnet PALA) og dens tetranatriumsalt er kendte forbindelser.This invention relates to a process for preparing the novel disodium salt of N- (phosphonoacetyl) -L-aspartic acid. The free tetravalent acid N- (phosphonoacetyl) -L-aspartic acid (hereinafter referred to in some cases as PALA) and its tetrasodium salt are known compounds.

Den kendte tetravalente N-(phosphonoacetyl)-L-asparaginsyre (PALA) blev først fremstillet af Stark et al., J. Biol. Chem., 246, 6599 (1971). Tetranatriumsal-10 tet af PALA er et kendt antitumormiddel som anført i litteraturen, specielt Cancer Research, 36, 2720 (1976).The known tetravalent N- (phosphonoacetyl) -L-aspartic acid (PALA) was first prepared by Stark et al., J. Biol. Chem., 246, 6599 (1971). The tetrasodium salt of PALA is a known antitumor agent as stated in the literature, especially Cancer Research, 36, 2720 (1976).

F. eks. blev overlevelsestiden for mus med intraperito-neal P388 leukæmi forlænget med op til 64% ved behandling med PALA-tetranatriumsalt i et dosisniveau mellem 15 188 og 750 mg/kg, i.p. Lewi's lungesarkoma var meget følsom overfor PALA-tetranatriumsalt hos mus ved i.p. doser fra 240 til 490 mg/kg. Mus med B16 melanoma overlevede 77 til 86% længere end kontroldyr, når der skete behandling med PALA-tetranatriumsalt (490 mg/kg, i.p.).For example, the survival time of mice with intraperitoneal P388 leukemia was extended by up to 64% by treatment with PALA tetrasodium salt at a dose level between 15 188 and 750 mg / kg, i.p. Lewi's pulmonary sarcoma was very sensitive to PALA tetrasodium salt in mice at i.p. doses from 240 to 490 mg / kg. Mice with B16 melanoma survived 77 to 86% longer than control animals when treated with PALA tetrasodium salt (490 mg / kg, i.p.).

20 Medens syntese af tetrasyren PALA ikke giver van skeligheder, har fremstillingen af tetranatriumsaltet, specielt i mængder i kilogramstørrelsen, vist sig at volde problemer, da udbytte og renhed for produktet er meget varierende. Fremgangsmåden ifølge opfindelsen er 25 særligt egnet til produktion af dinatriumsaltet i sådanne mængder og i ren form.While synthesis of the tetraic acid PALA does not cause difficulties, the preparation of the tetrasodium salt, especially in amounts in the kilogram size, has been found to cause problems, as the yield and purity of the product are very variable. The process of the invention is particularly suitable for producing the disodium salt in such amounts and in pure form.

Dinatriumsaltet af N- (phosi&onoacetyl) -L-asparaginsyre har formlen | jj'The disodium salt of N- (phosi & onoacetyl) -L-aspartic acid has the formula | jj '

NHCCH..P - OHNHCCH..P - OH

/ i 30 HOOC - CH2 - CH ONa ^COONa 35 Det hidtil ukendte dinatriumsalt af N-phosphono- acetyl)-L-asparaginsyre har ligesom det kendte PALA-tetranatriumsalt antitumorvirkning in vivo.The novel disodium salt of N-phosphonoacetyl) -L-aspartic acid, like the known PALA tetrasodium salt, has antitumor effect in vivo.

2 149134 PALA-dinatriumsaltet kan foreligge som et hydrat, som fra én præparation til en anden varierer med hensyn til indhold af hydratvand, typisk fra ca. 0,2-2 mol vand. Forbindelsen kan også som et solvat indeholde ethanol (f.The PALA disodium salt may be present as a hydrate which varies from one preparation to another with regard to the content of hydrate water, typically from 0.2-2 moles of water. The compound may also contain, as a solvate, ethanol (f.

5 eks. fra ca. 0,1 til 0,5 mol), eddikesyre (f.eks. fra ca. 0,03 til 0,4 mol) og natriumacetat (f.eks. ca. 0,2 mol). Eddikesyren og ethanolen kan fjernes ved frysetørring. 2-3 frysetørringer giver et opløsningsmiddel-frit materiale. Til anvendelse som antitumormiddel er 10 det hidtil ukendte dinatrium-PALA praktisk taget ækvivalent med det kendte tetranatrium-PALA med hensyn til aktivitet og toksicitet. Dinatrium-PALA kan anvendes til udnyttelse af dets antitumoraktivitet efter oparbejdning til farmaceutiske præparater, indeholdende en 15 forenelig farmaceutisk acceptabel bærer. Præparaterne kan også indeholde antimikrobielle midler og andre antitumormidler. Præparaterne kan oparbejdes i enhver farmaceutisk form, som er egnet til den påtænkte administreringsvej . Eksempler på sådanne præparater omfatter fa-20 ste præparater til oral administrering, såsom tabletter, kapsler, piller, pulvere og granulat, flydende præparater til lokal eller oral administrering, såsom opløsninger, suspensioner, saft og eliksirer og præparater til parenteral administrering, såsom sterile opløsninger, 25 suspensioner eller emulsioner.5 items from approx. 0.1 to 0.5 mole), acetic acid (e.g., from about 0.03 to 0.4 mole) and sodium acetate (e.g., about 0.2 mole). The acetic acid and ethanol can be removed by freeze-drying. 2-3 freeze dryers provide a solvent-free material. For use as an antitumor agent, the novel disodium PALA is practically equivalent to the known tetrasodium PALA in terms of activity and toxicity. Disodium PALA can be used to utilize its anti-tumor activity after processing into pharmaceutical compositions containing a compatible pharmaceutically acceptable carrier. The compositions may also contain antimicrobial agents and other antitumor agents. The compositions may be reprocessed in any pharmaceutical form suitable for the intended route of administration. Examples of such compositions include solid compositions for oral administration such as tablets, capsules, pills, powders and granules, liquid or local or oral preparations such as solutions, suspensions, juices and elixirs and parenteral administration preparations such as sterile solutions. , 25 suspensions or emulsions.

Til anvendelse som antitumormiddel administreres præparaterne i sådanne doser, at tumorvæksten hæmmes. Et foreslået dosisniveau til anvendelse som antitumormiddel (specielt til faste tumorer som beskrevet ovenfor) hos 30 pattedyr, er 50 til 500 mg dinatrium-PALA pr. kg legemsvægt til en enkelt daglig parenteral (f.eks. intravenøs infusion som en 2%'s vandig opløsning) behandling.For use as an antitumor agent, the compositions are administered in such doses as to inhibit tumor growth. A suggested dose level for use as an antitumor agent (especially for solid tumors as described above) in 30 mammals is 50 to 500 mg disodium PALA per day. kg of body weight for a single daily parenteral (eg intravenous infusion as a 2% aqueous solution) treatment.

Dinatrium-PALA er fordelagtigt ved at være et mobilt, fritstrømmende partikelformet stof, der let kan 35 håndteres, analyseres og vejes til formuleringsformål.Disodium PALA is advantageous in being a mobile, free-flowing particulate substance that can be easily handled, analyzed and weighed for formulation purposes.

Ved sammenligning med det kendte tetranatrium-PALA er det relativt ikke-hygroskopisk. Tetranatrium-PALA absorberer atmosfærisk fugtighed 1,5 gange hurtigere end 149134 3 dinatrium-PALA og er vanskeligt at håndtere og analysere. Dinatrium-PALA udmærker sig ved at have en opløselighed i vand, som er mindst 950 mg/ml. Det er således mere vandopløseligt end tetranatrium-PALA.When compared to the known tetrasodium PALA, it is relatively non-hygroscopic. Tetrasodium PALA absorbs atmospheric humidity 1.5 times faster than disodium PALA and is difficult to handle and analyze. Disodium PALA is distinguished by having a solubility in water of at least 950 mg / ml. Thus, it is more water soluble than tetrasodium PALA.

5 Følgende oplysninger tjener til identifikation af dinatrium-PALA: Dinatrium-PALA har som en 2 vægt/- volumen-% opløsning i vand en pH-værdi på ca. 4, sammenlignet med trinatrium-PALA og tetranatrium-PALA-opløs-ninger med pH-værdier på henholdsvis ca. 8 og ca. 9.5 The following information serves to identify disodium PALA: Disodium PALA has a pH value of approx. 2% w / v% solution in water. 4, compared with trisodium PALA and tetrasodium PALA solutions having pH values of approx. 8 and approx. 9th

10 Dinatrium-PALA er også karakteristisk ved at have et 60-MC kærnemagnetisk resonansspektrum (nmr), som typisk udviser en dublet svarende til methylengruppen (CH2), som er α-stillet til gruppen -CH, medens tetranatrium-PALA karakteristisk har et nmr-spektrum, som udviser en 15 treliniet multiplet svarende til den nævnte methylen-gruppe.Disodium PALA is also characterized by having a 60-MC nuclear magnetic resonance spectrum (nmr), which typically exhibits a doublet corresponding to the methylene group (CH spectrum which exhibits a triline multiple multiplied by the said methylene group.

Fremgangsmåden ifølge opfindelsen til fremstilling af dinatriumsaltet af N-(phosphonoacetyl)-L-aspara-ginsyre med formlen 20The process of the invention for the preparation of the disodium salt of N- (phosphonoacetyl) -L-aspartic acid of formula 20

O O .OHO O .OH

ir \ / ^IH-C-CH2-P^ HGOC-CH2-CH 0Na 2 5 COONa er ejendommelig ved det i kravets kendetegnende del anførte .The COONa is peculiar to the characterizing part of the claim.

Tetranatriumsaltet er hidtil fremstillet ved en 30 metode, hvor en først syntetiseret opløsning af den frie syre renses ved tids- og arbejdskrævende processer, bl. a. under anvendelse af ionbytningssøjler, derpå inddampes i vakuum, genopløses, titreres til pH ca. 9 og atter inddampes og tritureres gentagne gange.The tetrasodium salt has so far been prepared by a method in which a first synthesized solution of the free acid is purified by time and labor intensive processes, i. a. Using ion exchange columns which are then evaporated in vacuo, redissolved, titrated to pH ca. 9 and again evaporated and triturated repeatedly.

35 Fremstillingen af dinatrium-PALA ved fremgangsmå den ifølge opfindelsen medfører følgende fordele: (1) Der kræves kortere tid til syntese af materialet som følge af (a) fuldstændig eliminering af den 4 149134 tidligere anvendelse af ionbytningskolonner og (b) væsentlig formindskelse af den vandmængde, der skal bortdampes; (2) de beregnede omkostninger for en mængde af dinatri- 5 um-PALA på 5 kg er mindst 30% mindre end for 5 kg tetranatrium-PALA, fremstillet på sædvanlig måde; (3) den altid for lave hydrogenanalyse for tetranatri-um-JPALA, som rejser tvivl om, hvorvidt en given portion af stoffet faktisk er ren, er ikke længere 10 et problem; (4) syntesen er velegnet til gennemførelse i stor målestok; (5) det opnåede produkt er mindre hygroskopisk end tetranatrium-PALA, og i modsætning til sidstnævnte er 15 det et mobilt, fritstrømmende, partikelformet stof, som let kan håndteres og vejes med henblik på formulering, og (6) produktet har en meget stor opløselighed i vand.The preparation of disodium PALA by the method according to the invention has the following advantages: (1) Less time is required for the synthesis of the material as a result of (a) complete elimination of the prior use of ion exchange columns and (b) substantial reduction of the amount of water to be evaporated; (2) the calculated cost for an amount of 5 kg of disodium PALA of 5 kg is at least 30% less than that of 5 kg of tetrasodium PALA produced in the usual manner; (3) the always-too-low hydrogen analysis for tetrasodium-JPALA, which doubts whether a given portion of the substance is actually pure, is no longer a problem; (4) the synthesis is suitable for large-scale implementation; (5) the product obtained is less hygroscopic than tetrasodium PALA, and, in contrast to the latter, it is a mobile, free-flowing, particulate substance that can be easily handled and weighed for formulation, and (6) the product has a very large solubility in water.

Opfindelsen illustreres nærmere ved hjælp af føl-20 gende eksempel.The invention is further illustrated by the following example.

EKSEMPEL' L-Asparaginsyre-dibenzylester-p-toluensulfonat (I).Example 'L-Aspartic acid dibenzyl ester p-toluenesulfonate (I).

En omrørt blanding af 133 g, 1,00 mol, L-aspara-25 ginsyre, 650 g, 6,00 mol, benzylalkohol, 194 g, 1,02 mol, p-toluensulfonsyre-monohydrat og 400 ml tør benzen . blev opvarmet til tilbagesvaling i 6,5 timer. Det vand, der blev dannet ved reaktionen (46 ml), blev fjernet ved hjælp af en Dean-Stark-fælde. Den resulterende opløs-30 ning blev afkølet til stuetemperatur og derpå fortyndet med 400 ml benzen og 1200 ml ether. Ved afkøling til 10°C udskiltes et fast stof. Dette materiale blev opsamlet på et filter, vasket med vand, 1,0 1, og tørret, hvorved blev opnået 384 g, 79%, af det rå stof I. En 35 portion på 304 g af dette produkt blev omkrystalliseret fra 425 ml methanol til opnåelse af 186 g, (61,2%'s udvinding) af et materiale egnet til yderligere omdannelse, smp. 155,5-158°C; litteraturen angiver smp. 158 - 160°C.A stirred mixture of 133 g, 1.00 mol, L-aspartic acid, 650 g, 6.00 mol, benzyl alcohol, 194 g, 1.02 mol, p-toluenesulfonic acid monohydrate and 400 ml of dry benzene. was heated to reflux for 6.5 hours. The water formed by the reaction (46 ml) was removed by a Dean-Stark trap. The resulting solution was cooled to room temperature and then diluted with 400 ml of benzene and 1200 ml of ether. On cooling to 10 ° C, a solid is separated. This material was collected on a filter, washed with water, 1.0 L, and dried to yield 384 g, 79% of crude I. A 354 g portion of this product was recrystallized from 425 ml of methanol. to obtain 186 g, (61.2% recovery) of a material suitable for further conversion, m.p. 155.5 to 158 ° C; the literature indicates m.p. 158 - 160 ° C.

5 1491.345 1491.34

Phosphonoacetylchlorid (III) .Phosphonoacetyl chloride (III).

Til en omrørt blanding af 2000 g, 14,28 mol, phosphonoeddikesyre (II), 208,8 g, 2,856 mol, N,N-dime-thylformamid og 7,15 liter dioxan blev dråbevis sat 5 3568 g, 29,99 mol, thionylchlorid i løbet af 1,5 timer.To a stirred mixture of 2000 g, 14.28 mole, phosphonoacetic acid (II), 208.8 g, 2.856 mole, N, N-dimethylformamide and 7.15 liters of dioxane was added dropwise to 5,568 g, 29.99 mole. , thionyl chloride over 1.5 hours.

Temperaturen blev holdt under 30°C under tilsætningen.The temperature was kept below 30 ° C during the addition.

Den resulterende opløsning blev opvarmet til 45°C i 2\ time og derpå afkølet til 5°C. Derpå blev tilsat 283 ml, 15,7 mol, vand opløst i 2,5 liter dioxan dråbevis 10 i løbet af to timer. Temperaturen blev holdt under 10°C under tilsætningen. Denne opløsning af phosphonoacetylchlorid (III) blev omrørt ved 5-10°C i 40 minutter og derpå anvendt i nedenstående reaktion uden yderligere analyse.The resulting solution was heated to 45 ° C for 2 hours and then cooled to 5 ° C. Then, 283 ml, 15.7 mol, water dissolved in 2.5 liters of dioxane was added dropwise over two hours. The temperature was kept below 10 ° C during the addition. This solution of phosphonoacetyl chloride (III) was stirred at 5-10 ° C for 40 minutes and then used in the reaction below without further analysis.

15 L-Asparaginsyre-N-(phosphonoacetyl)-dibenzylester (IV).L-Aspartic acid N- (phosphonoacetyl) dibenzyl ester (IV).

En omrørt suspension af 4625 g, 9,525 mol, L-asparaginsyre-dibenzylester-p-toluensulfonat (I) i 20,0 liter dioxan blev afkølet til 15°C, hvorpå 4820 g, 47,63 mol, triethylamin blev tilsat som en svag strøm i 2Q løbet af en time. Den resulterende opløsning blev omrørt i 20 minutter og derpå blev tilsat den ovenfor beskrevne opløsning af phosphonoacetylchlorid (III), hvilken tilsætning skete dråbevis i løbet af fem timer. Temperaturen blev holdt under 20 C°under tilsætningen.A stirred suspension of 4625 g, 9.525 mol, L-aspartic acid dibenzyl ester p-toluenesulfonate (I) in 20.0 liters of dioxane was cooled to 15 ° C, to which 4820 g, 47.63 mol, triethylamine was added as a weak power in 2Q over an hour. The resulting solution was stirred for 20 minutes and then the above-described solution of phosphonoacetyl chloride (III) was added, which was added dropwise over five hours. The temperature was kept below 20 ° C during the addition.

25 1162 g, 11,48 mol, yderligere triethylamin blev tilsat og reaktionsblandingen omrørt i en time. Efter henstand i otte timer ved stuetemperatur blev blandingen fortyndet med 5,5 liter acetone og tørret i fem minutter, hvorpå uopløseligt materiale blev opsamlet på et filter og 30 vasket med 10,0 liter dioxan. En anden reaktion blev udført samtidig under de samme betingelser og under anvendelse af de samme mængder reagenser. Filtraterne fra de to forsøg blev forenet og inddampet i vakuum. Remanensen, som var en orange, viskos olie, blev opløst i 35 110,0 liter methylenchlorid, hvorpå den organiske fase blev vasket med vand og tørret over 11,3 kg natriumsulfat og 2,3 kg magnesiumsulfat. Uopløselige materialer blev filtreret fra (Celite-pude), og filtratet inddampet 6 1491341162 g, 11.48 mol, additional triethylamine was added and the reaction mixture stirred for one hour. After standing for eight hours at room temperature, the mixture was diluted with 5.5 liters of acetone and dried for five minutes, after which insoluble material was collected on a filter and washed with 10.0 liters of dioxane. Another reaction was carried out simultaneously under the same conditions and using the same amounts of reagents. The filtrates from the two experiments were combined and evaporated in vacuo. The residue, which was an orange viscous oil, was dissolved in 110.0 liters of methylene chloride, then the organic phase was washed with water and dried over 11.3 kg of sodium sulfate and 2.3 kg of magnesium sulfate. Insoluble materials were filtered off (Celite pad) and the filtrate evaporated 6 149134

i vakuum til konstant vægt. Udbytte af dibenzyl-PALAin vacuum to constant weight. Yield of dibenzyl-PALA

(IV) 7970 g (96,1%). Denne gule, viskose olie var egnet til yderligere omdannelse.(IV) 7970 g (96.1%). This yellow, viscous oil was suitable for further conversion.

L-Asparaginsyre-N-(phosphonoacetyl)-dibenzylester-cyclo-5 hexylaminsalt.L-Aspartic acid N- (phosphonoacetyl) dibenzyl ester cyclohexylamine salt.

1815 g, 18,30 mol, cyclohexylamin blev dråbevis sat til en kold (7°C) omrørt opløsning af 7970 g, 18,30 mol L-asparaginsyre-N-(phosphonoacetyl)-dibenzylester (IV) i 24,0 liter acetone i løbet af 1,25 timer. Tempe-10 raturen blev holdt under 15°C under tilsætningen. Kølebadet blev fjernet, og den resulterende opløsning omrørt i en time. Blandingen blev henstillet ved stuetemperatur i seks timer, hvorpå det udfældede faste stof blev opsamlet på et filter, vasket med 15,0 liter acetone og 15 tørret. Udbytte 4932 g, smp. 176,5 - 177,5°C. Dette stof blev omkrystalliseret fra 35,0 liter kogende methanol og derpå tørret til opnåelse af 1663 g af det rensede salt, smp. 178 - 181°C.1815 g, 18.30 mol, cyclohexylamine was added dropwise to a cold (7 ° C) stirred solution of 7970 g, 18.30 mol L-aspartic acid N- (phosphonoacetyl) dibenzyl ester (IV) in 24.0 liters of acetone within 1.25 hours. The temperature was kept below 15 ° C during the addition. The cooling bath was removed and the resulting solution stirred for one hour. The mixture was allowed to stand at room temperature for six hours, after which the precipitated solid was collected on a filter, washed with 15.0 liters of acetone and dried. Yield 4932 g, m.p. 176.5 - 177.5 ° C. This substance was recrystallized from 35.0 liters of boiling methanol and then dried to give 1663 g of the purified salt, m.p. 178 - 181 ° C.

Moderluden blev koncentreret i vakuum til et vo-20 lumen på 20,0 liter. Opløsningen blev fortyndet med 16,0 liter acetone og afkølet til -10°C til opnåelse af yderligere 967 g af produktet. Smp. 177 - 180°C. Et tredie udbytte på 429 g af materialet blev opnået ved at inddampe ovennævnte methanol-acetonefiltrat til omtrent-25 lig tørhed og suspendere remanensen i 5,0 liter acetone. Totalmængde af renset aminsalt egnet til yderligere omdannelse var 3059 g (62,0% udvinding).The mother liquor was concentrated in vacuo to a volume of 20.0 liters. The solution was diluted with 16.0 liters of acetone and cooled to -10 ° C to give an additional 967 g of the product. Mp. 177 - 180 ° C. A third yield of 429 g of the material was obtained by evaporating the above methanol-acetone filtrate to approximately 25 dryness and suspending the residue in 5.0 liters of acetone. Total amount of purified amine salt suitable for further conversion was 3059 g (62.0% recovery).

L-Asparaginsyre-N-(phosphonoacetyl)-tetranatriumsalt (V).L-Aspartic acid N- (phosphonoacetyl) tetrasodium salt (V).

Til en kold (5°C) omrørt opløsning af 1191 g, 30 32,28 mol, natriumhydroxid i 20,5 liter vand blev por tionsvis i løbet af 30 minutter sat 3059 g (svarende til 5,378 mol, hvis aminsaltet har samme empiriske formel som den analytisk rene prøve), L-asparaginsyre-N-(phosphonoacetyl)-dibenzylester-cyclohexylaminsalt.To a cold (5 ° C) stirred solution of 1191 g, 32.28 mole, sodium hydroxide in 20.5 liters of water was added 3059 g (305 g) in portion over 30 minutes, corresponding to the amine salt of the same empirical formula. as the analytically pure sample), L-aspartic acid N- (phosphonoacetyl) -dibenzyl ester cyclohexylamine salt.

35 Reaktionsblandingen blev omrørt ved 5-15°C i 3,5 timer og derpå ekstraheret med 2 x 8,5 liter methylen-chlorid og 1 x 8,5 liter ether. Den vandige opløsning blev klaret ved filtrering, koncentreret'i vakuum 149134 7 (<35°C, 3-5 mm Hg) til et volumen på 14,6 liter og derpå fortyndet med 51,4 liter ethanol. Den resulterende blanding blev omrørt i en time og henstillet ved stuetemperatur i 12 timer. Den vandige ethanolopløsning 5 blev fjernet efterladende rå (V) som en lysegul olie egnet til yderligere omdannelse.The reaction mixture was stirred at 5-15 ° C for 3.5 hours and then extracted with 2 x 8.5 liters of methylene chloride and 1 x 8.5 liters of ether. The aqueous solution was clarified by filtration, concentrated in vacuo (<35 ° C, 3-5 mm Hg) to a volume of 14.6 liters and then diluted with 51.4 liters of ethanol. The resulting mixture was stirred for one hour and allowed to stand at room temperature for 12 hours. The aqueous ethanol solution 5 was removed leaving crude (V) as a pale yellow oil suitable for further conversion.

L-Asparaginsyre-N-(phosphonoacetyl)-dinatriumsalt.L-aspartic acid-N- (phosphonoacetyl) disodium salt.

8,0 liter iseddikesyre blev sat til den ovenfor udskilte olie (rå asparaginsyre-N-(phosphonoacetyl)-te-10 tranatriumsalt (V) fremstillet ud fra 3059 g af aminsal-tet). Blandingen blev omrørt, hvorpå en gelatinøs udfældning blev filtreret fra. Det klare lysegule filtrat blev fortyndet med 24,0 liter ethanol. Den resulterende blanding blev omrørt i 1', 75 timer, hvorpå det udfæl-15 dede materiale blev opsamlet på et filter. Dette stof blev suspenderet i 14,5 liter ethanol, og blandingen omrørt kraftigt i en time. Produktet blev opsamlet på fire filtre og derpå partielt tørret i vakuum (30 - 45°C, aspiratortryk,derpå 3-5 mm Hg). Det klumpede materiale 20 (28,70 g) blev opløst i 5,25 liter vand, opløsningen blev klaret ved filtrering og filtratet (ca. 6,9 liter) fortyndet med 21,0 liter ethanol. Den'resulterende blanding blev omrørt i 30 minutter, hvorpå man lod den udskilte olie samle sig i en time. Den vandige ethanol-25 opløsning blev fjernet og olien vasket en gang med 4,3 liter ethanol. Dette materiale blev opløst i 8,15 liter vand, og opløsningen (9,8 liter) opdelt i tre portioner (to på 4,0 liter, en på 1,8 liter). Hver portion blev i løbet af 13 timer sat til den indre hvirvel i kraftigt 30 omrørt ethanol (10 gange volumenet af den vandige opløsning: 2 x 40,0 liter, 1 x 18,0 liter). Efter omrøring af blandingerne i to timer blev vand-ethanolopløsninger-ne fjernet, og det faste stof fra de tre udfældninger blev forenet. Dette materiale blev omrørt i 30 minut-35 ter i 10,0 liter ethanol, opsamlet på filter og derpå tørret til konstant vægt i vakuum ved stuetemperatur over phosphorpentaoxid. Det tørrede produkt (1748,0 g) blev passeret gennem en 150 μ rustfri stålsigte og bian- 8 149134 det omhyggeligt til opnåelse af dinatrium-PALA som et hvidt pulver.8.0 liters of glacial acetic acid was added to the above oil (crude aspartic acid N- (phosphonoacetyl) -tetrasodium salt (V) prepared from 3059 g of the amine salt). The mixture was stirred and a gelatinous precipitate was filtered off. The clear pale yellow filtrate was diluted with 24.0 liters of ethanol. The resulting mixture was stirred for 1 ', 75 hours, after which the precipitated material was collected on a filter. This substance was suspended in 14.5 liters of ethanol and the mixture stirred vigorously for one hour. The product was collected on four filters and then partially dried in vacuo (30 - 45 ° C, aspirator pressure, then 3-5 mm Hg). The bulky material 20 (28.70 g) was dissolved in 5.25 liters of water, the solution clarified by filtration and the filtrate (about 6.9 liters) diluted with 21.0 liters of ethanol. The resulting mixture was stirred for 30 minutes, then the separated oil was allowed to collect for one hour. The aqueous ethanol solution was removed and the oil washed once with 4.3 liters of ethanol. This material was dissolved in 8.15 liters of water and the solution (9.8 liters) divided into three portions (two of 4.0 liters, one of 1.8 liters). Each portion was added over 13 hours to the inner vortex in vigorously stirred ethanol (10 times the volume of the aqueous solution: 2 x 40.0 liters, 1 x 18.0 liters). After stirring the mixtures for two hours, the water-ethanol solutions were removed and the solid from the three precipitates was combined. This material was stirred for 30 minutes in 10.0 liters of ethanol, collected on filter and then dried to constant weight in vacuo at room temperature over phosphorus pentoxide. The dried product (1748.0 g) was passed through a 150 µ stainless steel screen and carefully mixed to obtain disodium PALA as a white powder.

Analyse:Analysis:

Beregnet for: '5 CgH^gNOgP - 2,4Na - 2 H20 - 0,5 C2HgO C 22,91, H 4,01, N 3,82, P 8,44, Na 15,04 Fundet: C 23,16, H 3,76, N 3,79, P 8,57, Na 15,18.Calcd for: 5 CgH2GNOgP - 2.4Na - 2H2O - 0.5 C2HgO C 22.91, H 4.01, N 3.82, P 8.44, Na 15.04 Found: C 23.16 , H 3.76, N 3.79, P 8.57, Na 15.18.

Natriumanalyse angiver en sammensætning svarende til:Sodium analysis indicates a composition similar to:

10 60% di-Na-PALA60% di-Na-PALA

40% tri-Na-PALA.40% tri-Na-PALA.

Baseret på den empiriske formel: % H20 = 9,8% % EtOH = 6,3%.Based on the empirical formula:% H 2 O = 9.8%% EtOH = 6.3%.

1515

Spektraldata: Kærnemagnetisk resonans (D20) δ 1,17 (t, 1,5, -CHg i ethanol), 2,74 (d, 2, **CH2 α til -CH), 2,77 (d, 2, J=20 Hz, -CH2 a til P), 3,63 (q, 1, 20 -CH2 i ethanol), 4,48 (t, 1, -CH)Spectral Data: Nuclear Magnetic Resonance (D 2 O) δ 1.17 (t, 1.5, -CH 2 in ethanol), 2.74 (d, 2, ** CH 2 α to -CH), 2.77 (d, 2, J = 20 Hz, -CH 2 a to P), 3.63 (q, 1, 20 -CH 2 in ethanol), 4.48 (t, 1, -CH)

Optisk drejning:Optical rotation:

Fundet + 14,73 (c, 2,098 i vand) 22Found + 14.73 (c, 2.098 in water) 22

Litteraturen angiver: [α]β + 14,86 (c, 1,998 i 25 vand).The literature indicates: [α] β + 14.86 (c, 1.998 in water).

Chromatografi:chromatography:

Tyndtlags chromatograf i 30 (Cellulose, Quanta/Gram Qf2 glasplader)Thin-layer chromatograph in 30 (Cellulose, Quanta / Gram Qf2 glass plates)

Opløsningsmiddelsystem R^-værdi 1. Lithiumchlorid (0,6 M)-ethanol- ammoniumhydroxid (5:5:1) 0,52 2. Ehtanol-vand (2:3) 0,72 35 3. Ethanol-ammoniumhydroxid- 0,16 vand (6:1:3) (aflang) 4. n-Butanol-eddikesyre-vand 0,22 (5:2:3) (kometformet)Solvent system R ^ value 1. Lithium chloride (0.6 M) -ethanol-ammonium hydroxide (5: 5: 1) 0.52 2. Ethanol-water (2: 3) 0.72 3. Ethanol-ammonium hydroxide-0, 16 water (6: 1: 3) (oblong) 4. n-Butanol acetic acid water 0.22 (5: 2: 3) (comet shaped)

DK506278A 1977-11-14 1978-11-14 METHOD FOR PREPARING THE DINATRUM SALT OF N- (PHOSPHONOACETYL) -L-ASPARAGINIC ACID DK149134C (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US85138277A 1977-11-14 1977-11-14
US85138277 1977-11-14

Publications (3)

Publication Number Publication Date
DK506278A DK506278A (en) 1979-05-15
DK149134B true DK149134B (en) 1986-02-10
DK149134C DK149134C (en) 1986-07-14

Family

ID=25310650

Family Applications (1)

Application Number Title Priority Date Filing Date
DK506278A DK149134C (en) 1977-11-14 1978-11-14 METHOD FOR PREPARING THE DINATRUM SALT OF N- (PHOSPHONOACETYL) -L-ASPARAGINIC ACID

Country Status (14)

Country Link
JP (1) JPS5492920A (en)
AT (1) AT374485B (en)
AU (1) AU520966B2 (en)
CA (1) CA1110266A (en)
CH (1) CH636595A5 (en)
DE (1) DE2849396A1 (en)
DK (1) DK149134C (en)
FI (1) FI65261C (en)
FR (1) FR2408617A1 (en)
GB (2) GB2008118B (en)
GR (1) GR65247B (en)
NL (1) NL7811252A (en)
NZ (1) NZ188909A (en)
SE (1) SE439774B (en)

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4284579A (en) * 1979-07-17 1981-08-18 The United States Of America As Represented By The Of The Department Of Health & Human Services (N-Phosphonacetyl-L-aspartato)(1,2-diaminocyclchexane)platinum(II) or alkali metal salt

Also Published As

Publication number Publication date
GR65247B (en) 1980-07-31
DK506278A (en) 1979-05-15
GB2008118B (en) 1982-08-04
ATA813278A (en) 1983-09-15
NL7811252A (en) 1979-05-16
FI65261B (en) 1983-12-30
AT374485B (en) 1984-04-25
SE439774B (en) 1985-07-01
FI65261C (en) 1984-04-10
GB2051070A (en) 1981-01-14
CA1110266A (en) 1981-10-06
DK149134C (en) 1986-07-14
DE2849396A1 (en) 1979-05-17
AU4152878A (en) 1979-05-24
GB2051070B (en) 1982-08-04
NZ188909A (en) 1980-10-08
GB2008118A (en) 1979-05-31
CH636595A5 (en) 1983-06-15
FR2408617A1 (en) 1979-06-08
AU520966B2 (en) 1982-03-11
FI783467A (en) 1979-05-15
JPS5492920A (en) 1979-07-23
SE7811736L (en) 1979-05-15
FR2408617B1 (en) 1983-11-18

Similar Documents

Publication Publication Date Title
EP0041355A1 (en) Novel erythromycin compounds
SU1375136A3 (en) Method of producing dioxipine or pyridoxine, or 2-guanidino-4-(2-methyl-4-imidazolyl)thiazole, or pyrebutyrole, or isoproterenole, or trimisosinic salt of pyroxycame
EP0003029B1 (en) New 2-oxo-1-pyrrolidine acetic acid derivatives, process for their preparation, medicaments containing them and process for their preparation
CN107286220B (en) 1,2, 4-triazole coupled dihydromyricetin derivative and preparation method and application thereof
CA1246062A (en) Anthracyclin derivatives, a process for their preparation and their use as cytostatic agents
CN111848690A (en) Anticancer tetravalent platinum complex capable of inhibiting inflammation and immune escape and preparation method thereof
DK149134B (en) METHOD FOR PREPARING THE DINATRUM SALT OF N- (PHOSPHONOACETYL) -L-ASPARAGINIC ACID
CN107382944B (en) Coumarin gossypol derivatives with anti-tumor activity and synthesis method thereof
US3812097A (en) Yranuronosyl)oxy)olean-12-en-30-oic acid
CN111333676A (en) Alkyl tin complex with antitumor activity and preparation method thereof
US2552896A (en) Methanol extraction of cascara sagrada bark
US5753695A (en) Flavilium compounds and method of using
EP2542560B1 (en) Arylfluorophosphate inhibitors of intestinal apical membrane sodium/phosphate co-transport
US4916220A (en) Palmitoyl-fructose-1,6-diphosphate, its therapeutic use and relevant preparation methods
EP0089532A2 (en) Substance with antiinflammatory and platelet antiaggregation activity extracted from plants of the family bromeliaceae or derived from bromelain and its extraction procedure
US4215070A (en) N-(Phosphonoacetyl)-L-aspartic acid compounds and methods for their preparation
US2929845A (en) Eulicinine
US4267126A (en) N-(Phosphonoacetyl)-L-aspartic acid compounds
CN112608327B (en) Furanoquinoline derivative, preparation method and application thereof
CN110818638B (en) Nitroxide free radical and preparation method and application thereof
EP0257488B1 (en) Novel diaminopyrimido[4,5-d]pyrimidine glycoside and glycotide and process
JPS6318590B2 (en)
EP0299413B1 (en) New tetramethyl-cis-diaza-bicyclo[4.2.0]octane-3,5-dione derivatives having a differentiation-inducing activity
CN110981897A (en) Zinc (II) complex, synthesis method and application thereof in antitumor drugs
JPH0689010B2 (en) Process for producing di-μ-hydroxoplatinum (II) binuclear complex nitrate

Legal Events

Date Code Title Description
PBP Patent lapsed