DK141910B - Process for the preparation of 1- (L - (-) - gamma-amino-alpha-hydroxybutyryl) -canamycin A or acid addition salts thereof. - Google Patents
Process for the preparation of 1- (L - (-) - gamma-amino-alpha-hydroxybutyryl) -canamycin A or acid addition salts thereof. Download PDFInfo
- Publication number
- DK141910B DK141910B DK59374AA DK59374A DK141910B DK 141910 B DK141910 B DK 141910B DK 59374A A DK59374A A DK 59374AA DK 59374 A DK59374 A DK 59374A DK 141910 B DK141910 B DK 141910B
- Authority
- DK
- Denmark
- Prior art keywords
- kanamycin
- compound
- water
- amino
- preparation
- Prior art date
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D209/00—Heterocyclic compounds containing five-membered rings, condensed with other rings, with one nitrogen atom as the only ring hetero atom
- C07D209/02—Heterocyclic compounds containing five-membered rings, condensed with other rings, with one nitrogen atom as the only ring hetero atom condensed with one carbocyclic ring
- C07D209/44—Iso-indoles; Hydrogenated iso-indoles
- C07D209/48—Iso-indoles; Hydrogenated iso-indoles with oxygen atoms in positions 1 and 3, e.g. phthalimide
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D207/00—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom
- C07D207/46—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with hetero atoms directly attached to the ring nitrogen atom
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H15/00—Compounds containing hydrocarbon or substituted hydrocarbon radicals directly attached to hetero atoms of saccharide radicals
- C07H15/20—Carbocyclic rings
- C07H15/22—Cyclohexane rings, substituted by nitrogen atoms
- C07H15/222—Cyclohexane rings substituted by at least two nitrogen atoms
- C07H15/226—Cyclohexane rings substituted by at least two nitrogen atoms with at least two saccharide radicals directly attached to the cyclohexane rings
- C07H15/234—Cyclohexane rings substituted by at least two nitrogen atoms with at least two saccharide radicals directly attached to the cyclohexane rings attached to non-adjacent ring carbon atoms of the cyclohexane rings, e.g. kanamycins, tobramycin, nebramycin, gentamicin A2
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C2603/00—Systems containing at least three condensed rings
- C07C2603/02—Ortho- or ortho- and peri-condensed systems
- C07C2603/04—Ortho- or ortho- and peri-condensed systems containing three rings
- C07C2603/22—Ortho- or ortho- and peri-condensed systems containing three rings containing only six-membered rings
- C07C2603/26—Phenanthrenes; Hydrogenated phenanthrenes
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Biotechnology (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Molecular Biology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Saccharide Compounds (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
Description
(11) FREMLÆGGELSESSKRIFT 1 4 1 9 1 O(11) PRESENTATION 1 4 1 9 1 O
(S) \RR/ DANMARK <» mtci.’ o 07 h 16/22 • (21) Antegning nr. 593/74 (22) Indlaveret den 4, fet). 1974 (23) Lgbedag 4. fet). 197^ (44) Antegningen fremlagt og -,οπ * fremlæggelaeetkriftet offentliggjort den 14. JUl · 1(S) \ RR / DENMARK <»mtci. 'O 07 h 16/22 • (21) Note No. 593/74 (22) Inserted on 4, bold). 1974 (23) Lgbedag 4. bold). 197 ^ (44) The note presented and -, οπ * the submission document published on 14 July · 1
Dl REKTORATET FORDl THE RECTORATE FOR
PATENT-OG VAREMÆRKEVÆSENET (30) Morittt begæret fra denPATENT AND TRADE MARKET (30) Morittt requested from it
7. feb. 1973, 330377, USFeb 7 1973, 330377 US
(71) BRISTOL-MYERS COMPANY, 545 Park Avenue, New York, N.Y. 10022, US.(71) BRISTOL-MYERS COMPANY, 545 Park Avenue, New York, N.Y. 10022, US.
(72) Opfinders Richard H. Schrelber, R.D.No.4, Beebe Bridge Road,(72) Inventors Richard H. Schrelber, R.D.No.4, Beebe Bridge Road,
Canastota, New York, “US: John G. Kell, 4504 Limestone Drive, Manlius,Canastota, New York, US: John G. Kell, 4504 Limestone Drive, Manlius,
New York, US.New York, US.
(74) Fuldmægtig under tagene behandling:(74) Clerk under the roofs processing:
Th. Ogtenf'eld Patent bur eau A/s.__ (64) Fremgangsmåde til fremstilling af 1 -(L-(-)-gamma-amino-alfa-hydroxy* butyryl)-kanamycin A eller syreadditionssalte deraf.Th. (64) Process for the preparation of 1- (L - (-) - gamma-amino-alpha-hydroxy-butyryl) -canamycin A or acid addition salts thereof.
Den foreliggende opfindelse angår en særlig fremgangsmåde til fremstilling af 1-[L-(-)-γ-amino-a-hydroxybutyryl]-kanamycin A (IV), der er en kendt forbindelse med formlen: 141910 2 • 6« 4, ck2nh2The present invention relates to a particular process for the preparation of 1- [L - (-) - γ-amino-α-hydroxybutyryl] -canamycin A (IV), a known compound of the formula: ck2nh2
HO-k IHO-k I
/2 NAi' H0 Λ h NHp °— ?%0H H0^~7£Al™ H0 >1^ / H2N 3" 2" Γ^ν/ ΉΟ-CH : I / ch2 H0 0 ^-5¾ eller et farmakologisk acceptabelt syreadditionssalt deraf, ud fra kanamycin A./ 2 NAi 'H0 Λ h NHp ° -?% 0H H0 ^ ~ 7 £ Al ™ H0> 1 ^ / H2N 3 "2" Γ ^ ν / ΉΟ-CH: I / ch 2 H0 0 ^ -5¾ or a pharmacologically acceptable acid addition salt thereof, from kanamycin A.
Kanamycin A, som er et kendt antibiotikum beskrevet i Merck Index, 8. udgave, side 597-598, har formlen 6* CH3-NH9Kanamycin A, a known antibiotic described in the Merck Index, 8th edition, pages 597-598, has the formula 6 * CH3-NH9
H°-K I HOH ° -K I HO
' H0 Λ1' lx nh2 6" VsLlo CHpOH ε HOA-_Λ H0 ^Kj5 - /2.iN\ !"/"H0 Λ1" lx nh2 6 "VsLlo CHpOH ε HOA-_Λ H0 ^ Kj5 - /2.iN \!" /
H0 HH0 H
Det ses, at kanamycin A besidder fire primære aminfunktioner ved 1-, 3-, 6'- og 3"-stillingerne i molekylet. Det er fastslået, at 6'-aminfunktionen er den mest reaktive, og at 1-aminfunktionen er den funktion, som har den næsthøjeste reaktivitet ved behandling med et elektrofilt middel. Aminfunktionerne ved 3- og 3"-stillingerne 141910 3 er mindre reaktive end aminfunktionerne ved 1- og 61-stillingerne, men reagerer ved acylering til dannelse af små mængder uønskede acylerede materialer.It is seen that kanamycin A possesses four primary amine functions at the 1-, 3-, 6'- and 3 "positions of the molecule. It has been established that the 6'-amine function is the most reactive and that the 1-amine function is the function. The amine functions at the 3 and 3 "positions 141910 3 are less reactive than the amine functions at the 1- and 61 positions, but react by acylation to form small amounts of undesired acylated materials.
Fra beskrivelsen til dansk patentansøgning nr. 3461/72 er det kendt at fremstille 1-[L- (»·)-γ-amino-a-hydroxybutyryl]-kanamycin A eller B ved at man blokerer aminogruppen på 61-carbønatomet i kanamycin A eller B, derefter selektivt acylerer aminogruppen ved 1-stillingen med et derivat af L-(-)-Y-amino-a-hydroxysmørsyre, hvori aminogruppen er beskyttet, og endelig fjerner de beskyttende grupper fra såvel 6’-stillingen som substituenten i 1-stillingen. Når denne kendte fremgangsmåde anvendes til fremstilling af kanamycin A derivatet (IV), kan der på grundlag af eksempel 3 i den nævnte ansøgning beregnes et totaludbytte på ca. 10% i forhold til den anvendte mængde af udgangsmaterialet kanamycin A.From the specification of Danish Patent Application No. 3461/72, it is known to prepare 1- [L- (R) -γ-amino-α-hydroxybutyryl] -canamycin A or B by blocking the amino group of the 61-carbon atom of kanamycin A or B, then selectively acylates the amino group at the 1-position with a derivative of L - (-) - γ-amino-α-hydroxybutyric acid in which the amino group is protected, and finally removes the protecting groups from both the 6 'position and the substituent in 1 position. When this known method is used to prepare the kanamycin A derivative (IV), on the basis of Example 3 of the said application, a total yield of approx. 10% relative to the amount of the starting material used kanamycin A.
Den foreliggende opfindelse tilvejebringer en forbedret fremgangsmåde til fremstilling af den omhandlede forbindelse (IV) ud fra kanamycin A, hvorved man opnår et væsentligt højere udbytte, jfr. efterfølgende eksempel 3, hvor udbyttet var 19,2%.The present invention provides an improved process for preparing the subject compound (IV) from kanamycin A, thereby obtaining a substantially higher yield, cf. following Example 3, where the yield was 19.2%.
Dette er ifølge opfindelsen opnået ved fremgangsmåden, der er ejendommelig ved det i kravets kendetegnende del anførte. Denne fremgangsmåde består i almindelighed i, at man først omsætter et molækvivalent kanamycin A med højst et molækvivalent N-(bénzyloxycarbonyloxy)-succinimid i et opløsningsmiddel. Som opløsningsmiddel anvendes fortrinsvis dimethylformamid, dimethylacetamid, tetrahydrofuran, dloxan, 1,2-dimethoxyethan, methanol, ethanol, vand, acetone, pyridin, en N-alkylpiperidin, hvor alkylgruppen kan være lige eller forgrenet og indeholder fra 1 til 6 carbonatomer eller blandinger deraf, især dimethylformamid. Omsætningen sker ved en temperatur under 50°C, fortrinsvis under 25°C, og især fra ca. -20°C til ca. 10°C. Der dannes herved en forbindelse med formlen: 141910 4 Η 0This is achieved according to the invention by the method which is characterized by the characterizing part of the claim. This process generally consists in first reacting a molar equivalent of kanamycin A with at most one molar equivalent of N- (benzyloxycarbonyloxy) succinimide in a solvent. Preferably, as the solvent, dimethylformamide, dimethylacetamide, tetrahydrofuran, dloxane, 1,2-dimethoxyethane, methanol, ethanol, water, acetone, pyridine, an N , especially dimethylformamide. The reaction is carried out at a temperature below 50 ° C, preferably below 25 ° C, and especially from ca. -20 ° C to approx. 10 ° C. A compound of the formula is thus formed: 141910 4 Η 0
Η IIΗ II
ho—ν ch2-n-c-o-ch2-c6h5 Η0_\^^-0 ' no \λ \>-- ΝΗ2 V-Λ ηο—^-ΤΧΛ /^Λ_νη2 ΗΟ—ν CH2OH /ho-ν ch2-n-c-o-ch2-c6h5 Η0 _ \ ^^ - 0 'no \ λ \> - ΝΗ2 V-Λ ηο - ^ - ΤΧΛ / ^ Λ_νη2 ΗΟ — ν CH2OH /
/ II/ II
Et mplækvivalent af forbindelse II behandles dernæst med mindst 3, fortrinsvis 3-6 og især 3,5 til 4,5,molækvivalenter 4-nitrobenz-aldehyd/ benzaldehyd, 4-methoxybenzaldehyd eller trimethylacetaldehyd, fortrinsvis p-nitrobenzaldehyd eller benzaldehyd, især p-nitrobenzalde-hyd, i en monofunktionel alkohol med lige eller forgrenet carbonkæde og indeholdende 1-6 carbonatomer, fortrinsvis absolut ethanol, methanol, η-propanol, isopropanol, n-butanol, sek-butanol, tert-butanol, især absolut ethanol eller blandinger deraf, ved forhøjet temperatur, fortrinsvis i intervallet fra 50°C til tilbagesvalingstemperaturen for systemet, især ved tilbagesvalingstemperaturen, i mindst 2 timer, fortrinsvis fra 2 til 10 timer, især fra 2 til 4 timer, til dannelse af forbindelsen med formlen: 5 1A1910 H ? ch2-n-c-o-ch2-c6h5 >ί-Λ ηο.Λ— \A mp equivalent of compound II is then treated with at least 3, preferably 3-6 and most preferably 3.5 to 4.5, molar equivalents of 4-nitrobenz-aldehyde / benzaldehyde, 4-methoxybenzaldehyde or trimethylacetaldehyde, preferably p-nitrobenzaldehyde or benzaldehyde, especially p. nitrobenzaldehyde, in a monofunctional alcohol of straight or branched carbon chain and containing 1-6 carbon atoms, preferably absolute ethanol, methanol, η-propanol, isopropanol, n-butanol, sec-butanol, tert-butanol, especially absolute ethanol or mixtures thereof , at elevated temperature, preferably in the range of 50 ° C to the reflux temperature of the system, especially at reflux temperature, for at least 2 hours, preferably from 2 to 10 hours, especially from 2 to 4 hours, to form the compound of the formula: ? ch2-n-c-o-ch2-c6h5> ί-Λ ηο.Λ— \
. / —Z. / —Z
HO__ CH-,ΟΗ / 1 ·%ν 0 111 Η H _ H ,-> hvor Z betegner -N=C-^ ^-Ν03 ’ \___y ’ ^-OCHg H ch3 eller -N=C-C-CH,.HO__ CH-, ΟΗ / 1 ·% ν 0 111 Η H _ H, -> where Z represents -N = C- ^ ^ -Ν03 '\ ___ y' ^ -OCHg H ch3 or -N = C-C-CH,.
I 3 CH3I 3 CH3
Et molækvivalent af forbindelse III behandles derefter med mindst 1, fortrinsvis 1-1,3, især 1,1 - 1,3 molækvivalent L-(-)-Y-benzyl-oxycarbonylamino-α-hydroxysmørsyre N-hydroxysuccinimidester i et organisk opløsningsmiddel, fortrinsvis dimethylformamid, tetrahydrofuran, dimethylacetamid, propylenglycoldimethylether, ethylenglycoldimethyl-ether eller dioxan, især dlmethylformamid, ved en temperatur i intervallet fra 0°C til ca. 50°C, fortrinsvis fra ca.10°C til ca. 35°C, især stuetemperatur, fortrinsvis i mindst 5 timer, hvorefter man fjerner opløsningsmidlet, fortrinsvis i vakuum, og hydrogenerer remanensen, fortrinsvis in situ, med hydrogen i nærværelse af palladium, platin, rhodium, Raney-nikkel, ruthenium eller nikkel, især palladium-på-trækul, i et vand-vand blandbart opløsningsmiddelsystem, fortrinsvis vand og dioxan, tetrahydrofuran eller ethylenglycoldimethylether, især en 1:1 blanding af dioxan og vand, fortrinsvis i nærværelse af en katalytisk mængde iseddikesyre, til dannelse af forbindelsen IV, som om ønsket omdannes til et farmaceutisk acceptabelt syreadditionssalt deraf.A mole equivalent of compound III is then treated with at least 1, preferably 1-1.3, especially 1.1 - 1.3 mole equivalent of L - (-) - Y-benzyl-oxycarbonylamino-α-hydroxybutyric acid N-hydroxysuccinimide ester in an organic solvent. preferably dimethylformamide, tetrahydrofuran, dimethylacetamide, propylene glycol dimethyl ether, ethylene glycol dimethyl ether or dioxane, especially dimethylformamide, at a temperature in the range of from 0 ° C to approx. 50 ° C, preferably from about 10 ° C to approx. 35 ° C, especially at room temperature, preferably for at least 5 hours, then removing the solvent, preferably in vacuo, and hydrogenating the residue, preferably in situ, with hydrogen in the presence of palladium, platinum, rhodium, Raney nickel, ruthenium or nickel, especially palladium-on-charcoal, in a water-water miscible solvent system, preferably water and dioxane, tetrahydrofuran or ethylene glycol dimethyl ether, in particular a 1: 1 mixture of dioxane and water, preferably in the presence of a catalytic amount of glacial acetic acid, to give the compound IV, which if desired, converted to a pharmaceutically acceptable acid addition salt thereof.
141910 6141910 6
Forbindelse IV, 1-[L-(-)-γ-amino-a-hydroxybutyryl]-kanamycin A besidder fremragende antibakteriel aktivitet, som viser sig at være aktiviteten af kanamycin A selv overlegen. Nedenfor findes to tabeller, som viser de minimalt inhibitoriske koncentrationer (MIC) af kanamycin A og forbindelse IV (BB-K8) mod et antal forskellige grampositive og gram-negative bakterier, hvilke MlC-værdier tilvejebragtes ved hjælp af Steers agar-fortyndingsmetode (tabel I) og den to-foldige fortyndingsmetode (tabel II). Kueller-Hinton agar-medium anvendtes ved de til grund for tabel I liggende undersøgelser, og Heart Infusion Broth anvendtes ved de til grund for tabel II liggende undersøgelser.Compound IV, 1- [L - (-) - γ-amino-α-hydroxybutyryl] -canamycin A possesses excellent antibacterial activity which is found to be the activity of kanamycin A itself superior. Below are two tables showing the minimal inhibitory concentrations (MIC) of kanamycin A and compound IV (BB-K8) against a number of different Gram-positive and Gram-negative bacteria, which M1C values were obtained by Steer's agar dilution method (Table I) and the two-fold dilution method (Table II). Kueller-Hinton agar medium was used in the Table I studies and Heart Infusion Broth was used in the Table II studies.
Tabel ITable I
_MIC mg/ml__MIC mg / ml_
Forbindelse IV Kanamycin A (BB-K8)Compound IV Kanamycin A (BB-K8)
Organisme (6-8198) Frøve nr. 4 1. Alk. faecalis A-9423 16 8 2. " " A-20648 >125 >125 3. Ent. cloacae A-0656 4 4 4. " species A-20364 >125 2 5. " hafniae 1 A-20674 1 1 6. Ξ. coli A-0636 2 1 7. M " A-20664 16 4 8. " " A-20665 >125 1 9. " " A-20507 32 2 10. " " A-20520 >125 4 11. ” ” A-20365 >125 1 12. " " A-20684 2 2 13. " " A-20682 >125 2 14. ” " A-20683 >125 8 15. " " A-20681 >125 2 16. " " A-15119 4 4 17. K. pneumoniae A-0967 4 4 18. " species A-20328 >125 2 19. " " A-20330 32 32 141910 7Organism (6-8198) Sample No. 4 1. Alk. faecalis A-9423 16 8 2. "" A-20648> 125> 125 3. Ent. cloacae A-0656 4 4 4. "species A-20364> 125 2 5." hafniae 1 A-20674 1 1 6. Ξ. coli A-0636 2 1 7. M "A-20664 16 4 8." "A-20665> 125 1 9." "A-20507 32 2 10." "A-20520> 125 4 11." "A- 20365> 125 1 12. "" A-20684 2 2 13. "" A-20682> 125 2 14. "" A-20683> 125 8 15. "" A-20681> 125 2 16. "" A-15119 4 4 17. K. pneumoniae A-0967 4 4 18. "species A-20328> 125 2 19." "A-20330 32 32 141910 7
Tabel I (fortsat) _MIC mg/ml_Table I (continued) _MIC mg / ml_
Forbindelse IV Kanamycin A (BB-K8)Compound IV Kanamycin A (BB-K8)
Organisme (6-8198) Prøve nr. 4 20. K. species A-20634 >125 4 21. " pneumoniae A-20680 >125 4 22. " " A-0077 1 1 23. Pr. mirabilis A-9900 2 2 24. Pr. morganii A-15153 2 2 25. " vulgaris A-9555 2 1 26. " rettgeri A-9636 0,25 0,25 27. " mirabilis A-20645 4 4 28. " " A-20454 2 2 29. Providencia stuartii A-20615 2 1 30. " alkalifaciens A-20676 1 1 31. Ps. aeruginosa A-20229 32 2 32. " " A-0943A 125 16 33. " " A-20653 >125 32 34 " species A-20601 125, 63 16 35. " " A-20621 >125 >125 36. " maltophilia A-20620 32 >125 37. Sal. enteritidis A-0531 1 0,5 38. " derby A-20087 >125 1 39. Ser. marcescens A-20019 2 4 40. " " A-9933 4 8 41. " " A-20460 >125 4, 2 42. " " A-20459 4 16 43. Shig. flexneri A-9684 4 4 44. Aeromonas sp. A-20670 2 2 45. Arizona sp. A-20671 2 1 46. Citrobacter sp. A-20673 4 4 47. Edwardsiella sp. A-20678 4 4 48. Staph, aureus A-9606 1 1 49. " " A-4749 0,5 i 50. " " A-9537 2 1 51. " " A-20610 >125 2 52. " " A-20240 >125 8 53. " " A-15197 1 2 141910 8Organism (6-8198) Specimen No. 4 20. K. species A-20634> 125 4 21. "pneumoniae A-20680> 125 4 22." "A-0077 1 1 23. Per mirabilis A-9900 2 2 24. Per morganii A-15153 2 2 25. "vulgaris A-9555 2 1 26." Straightening A-9636 0.25 0.25 27. "mirabilis A-20645 4 4 28." "A-20454 2 2 29. Providencia stuartii A-20615 2 1 30. "Alkalifaciens A-20676 1 1 31. Ps. Aeruginosa A-20229 32 2 32." "A-0943A 125 16 33." "A-20653> 125 32 34" species A-20601 125, 63 16 35. "" A-20621> 125> 125 36. "maltophilia A-20620 32> 125 37. Sal. Enteritidis A-0531 1 0.5 38." derby A-20087> 125 1 39. Ser. marcescens A-20019 2 4 40. "" A-9933 4 8 41. "" A-20460> 125 4, 2 42. "" A-20459 4 16 43. Shig. flexneri A-9684 4 4 44. Aeromonas sp. A-20670 2 2 45. Arizona sp. A-20671 2 1 46. Citrobacter sp. A-20673 4 4 47. Edwardsiella sp. A-20678 4 4 48. Staph, aureus A-9606 1 1 49. "" A-4749 0.5 i 50. "" A-9537 2 1 51. "" A-20610> 125 2 52. "" A -20240> 125 8 53. "" A-15197 1 2 141910 8
Tabel I (fortsat) _MIC mg/ml_Table I (continued) _MIC mg / ml_
Forbindelse IV Kanamycin A (BB-K8)Compound IV Kanamycin A (BB-K8)
Organisme (6-8198) Prøve nr. 4Organism (6-8198) Sample # 4
Mueller-Hinton medium + 4¾ fåreblod 54. Str. faecalis A-9854 63 63 55. " " A-9575 125 >125 56. Str. pyogenes A-20200 32, 16 32 57. " " A-9604 125 125 58 " " A-15040 125 125 59. " " A-20065 125 125 60. D. pneumoniae A-9585 63, 32 63 61. " " A-20159 125 >125Mueller-Hinton medium + 4¾ sheep blood 54th Str. faecalis A-9854 63 63 55. "" A-9575 125> 125 56. Str. pyogenes A-20200 32, 16 32 57. "" A-9604 125 125 58 "" A-15040 125 125 59. "" A-20065 125 125 60. D. pneumoniae A-9585 63, 32 63 61. "" A-20159 125> 125
Tabel IITable II
_MIC mg/ml__MIC mg / ml_
Forbindelse IV Kanamycin A (BB-K8)Compound IV Kanamycin A (BB-K8)
Organisme (6-8196) Prøve nr. 4 1. D. pneumoniae ±5% serum A-0585 63 63 2. Str. pyrogenes ±5% serum A-9604 125 125 3. Staph, aureus Smith A-9537 0,5 0,5 4. Staph, aureus A-9497 0,5 0,5 5. Staph, aureus A-20239 125 4 6. Staph, aureus A-20240 125 4 7. Enter, cloacae A-0656 2 2 8. Enter, species A-20364 125 2 9. K. pneumoniae A-9867 2 4 10. E. coli K-12 ML1410 A-20361 2 4 11. E. coli K-12 ML1630 A-20363 125 2 12. E. coli K-12 A-9632 2 1 13. E. coli A-20664 32 8 14. E. coli A-20665 125 8 15. Pr. mirabilis A-9900 2 16 16. Pr. morganii A-15153 4 16 17. Pr. vulgaris A-9436 1 2 18. Ps. aeruginosa A-20227 4 1 19. Ps. species A-20499 63 4 20. Ps. aeruginosa A-20653 125 4 141910 9Organism (6-8196) Sample No. 4 1. D. pneumoniae ± 5% serum A-0585 63 63 2. Size. pyrogenes ± 5% serum A-9604 125 125 3. Staph, aureus Smith A-9537 0.5 0.5 4. Staph, aureus A-9497 0.5 0.5 5. Staph, aureus A-20239 125 4 6 Staph, aureus A-20240 125 4 7. Enter, cloacae A-0656 2 2 8. Enter, species A-20364 125 2 9. K. pneumoniae A-9867 2 4 10. E. coli K-12 ML1410 A- 20361 2 4 11. E. coli K-12 ML1630 A-20363 125 2 12. E. coli K-12 A-9632 2 1 13. E. coli A-20664 32 8 14. E. coli A-20665 125 8 15. Pr. mirabilis A-9900 2 16 16. Pr. morganii A-15153 4 16 17. Pr. vulgaris A-9436 1 2 18. Ps. aeruginosa A-20227 4 1 19. Ps. species A-20499 63 4 20. Ps. aeruginosa A-20653 125 4 141910 9
Tabel II (fortsat) MIC mg/ml _Table II (continued) MIC mg / ml
1 ~ ' F6ft)in<iélse IV1 ~ 'F6ft) in <all IV
Kanamycin A (BB-K8)Kanamycin A (BB-K8)
Organisme (6-8196) Prøve nr. 4 21. Ps. species A-20621 125 125 22. Ser. marcescens A-20019 2 4 23. Ser. marcescens A-20141 16 16Organism (6-8196) Sample No. 4 21. Ps. species A-20621 125 125 22. Ser. marcescens A-20019 2 4 23. Ser. marcescens A-20141 16 16
De ovenfor anførte MIC-værdier viser, at forbindelse IV (BB-K8) er kanamycin A overlegen med hensyn til aktivitet overfor de fleste mikroorganismer, især ved kanamycin A resistente organismer.The above MIC values show that compound IV (BB-K8) is kanamycin A superior in activity to most microorganisms, especially kanamycin A resistant organisms.
MIC-værdierne stemte også godt overens med de resultater, som opnåedes ved afprøvning af kanamycin A og forbindelse IV in vivo mod nedennævnte tre forskellige organismer.The MIC values also corresponded well with the results obtained in testing kanamycin A and compound IV in vivo against the three different organisms listed below.
Forbindelse IV og kanamycin A var lige effektive ved infektioner hos mus forårsaget af kanamycin A-følsomme stammer af E. coli A-1519 og Staph, aureus A-9537. Skønt CD^q-værdierne (helbredende dosis for 50% af letalt inficerede must) for Staph, aureus A-9537 antyder, at forbindelse IV er en smule mindre aktiv end kanamycin A, er denne lille forskel sandsynligvis ikke signifikant, fordi dosisniveauerne lå langt fra hinanden (5 x fortynding).Compound IV and kanamycin A were equally effective in infections in mice caused by kanamycin A-sensitive strains of E. coli A-1519 and Staph, aureus A-9537. Although the CD ^ q values (healing dose for 50% of lethally infected must) for Staph, aureus A-9537 suggest that compound IV is slightly less active than kanamycin A, this small difference is probably not significant because dose levels were far apart (5 x dilution).
Kanamycin A var som ventet ikke særlig effektiv in vivo mod den kanamycin-resistente stamme af E. coll A-20520, hvorimod forbindelse IV udviste en tydelig beskyttende virkning. Forbindelse IV viste sig ca. 10 gange mere aktiv overfor denne E. coli-stamme ved indgivelse i en 4-behandlingskur end ved indgivelse i en 2-behandlings-kur.As expected, kanamycin A was not very effective in vivo against the kanamycin-resistant strain of E.coll A-20520, whereas compound IV showed a distinct protective effect. Compound IV was found to be ca. 10 times more active against this E. coli strain when administered in a 4-treatment regimen than by administration in a 2-treatment regimen.
1A1910 10 rH C <d cd o υ p to μ ·η1A1910 10 rH C <d cd o υ p to μ · η
P pi 0 0 PP pi 0 0 P
•Η N "Cf od Ή- 0 ·Η fl P (¾ rH o S3 ® Ή O i-n X X XX £> o> M-j ø Q rH P 'O t»• Η N "Cf od Ή- 0 · Η fl P (¾ rH o S3 ® Ή O i-n X X XX £> o> M-j ø Q rH P 'O t»
C_j o lo o o o 0 3 PC_j o lo o o o 0 3 P
cd o vO O O Λ i—I cd 0 Pcd o vO O O Λ i — I cd 0 P
•ri od N N 1/1 Η 43 £ ^ LO Ή O 0 *rl r* øo 43 3 4b p ø• ri od N N 1/1 Η 43 £ ^ LO Ή O 0 * rl r * øo 43 3 4b p ø
•HN u S P• HN u S P
M L I to VO rXM L I to VO rX
o 0<£ CM CH 3 to 43 O lo 3 3 S bo 0 b> Ø 1—1 N 1 CM I O 4b j_i ø S 1-1 pø · m 0 tO > 't £o 0 <£ CM CH 3 to 43 O lo 3 3 S bo 0 b> Ø 1—1 N 1 CM I O 4b j_i ø S 1-1 p · m 0 tO> 't £
0 > rH p O0> rH p O
w 0 0 0 » tOw 0 0 0 »tO
m Xi bo cd 0 rX 3 3 3 rø -H tO 0 -H » 0 3 pH O Od Od 0> rH O > • H O CO .P 3 ©- Æ U Q XI XI _ ^ 5¾5¾ M CJ g cd O Pi O cd cn od OP 43 iM-i-it-r to 0 ø pj to 43 pH Ό 43 0 0 μ υ co to fd p pm Xi bo cd 0 rX 3 3 3 rø -H tO 0 -H »0 3 pH O Od Od 0> rH O> • HO CO .P 3 © - Æ UQ XI XI _ ^ 5¾5¾ M CJ g cd O Pi O cd cn od OP 43 iM-i-it-r to 0 ø pj to 43 pH Ό 43 0 0 μ υ co to fd pp
O Mrl 0 Pi cm 35 PO Mrl 0 Pi cm 35 P
n_i p I P Cd tX Ό.n_i p I P Cd tX Ό.
0 <£ P r~\ tO Cm M 43 CJ ©. O P bO Xi ø u>—i od i ·3- i cm o^ ø 3 3 p tfl s X) Od rH ·γΗ ørn 3 rH rH t-H p 43 cd Ό 00 <£ P r ~ \ tO Cm M 43 CJ ©. O P bO Xi ø u> —i od i · 3- in cm o ^ ø 3 3 p tfl s X) Od rH · γΗ eagle 3 rH rH t-H p 43 cd Ό 0
•H p - P 3 XI• H p - P 3 XI
·> DO· C J) C·> DO · C J) C
H > ** Cd 43 ø +J CO N Od "©.Od 00 44 3 p P X 4b to ι-ted øoxi xi eu· 'i i—i M lo ft bO od 0 HH O 3QO LO · rX 43 t> 3 CJ Λ Λ '—v Λ ^— Xi pH ·Μ OJ O t-H Ød bo øøH> ** Cd 43 ø + J CO N Od "© .Od 00 44 3 p PX 4b to ι-ted øoxy xi eu · 'ii — i M lo ft bO od 0 HH O 3QO LO · rX 43 t> 3 CJ Λ Λ '—v Λ ^ - Xi pH · Μ OJ O tH Desolate island
0 >. to £ tO £ r* P0>. to £ tO £ r * P
43 3 Ο- X. Od '—' Ό cd 3 u Μ Μ H 33· Η·η ø lo £ øOcd/Ό O σι '—' X 3 Ή σι43 3 Ο- X. Od '-' Ό cd 3 u Μ Μ H 33 · Η · η ø lo £ øOcd / Ό O σι '-' X 3 Ή σι
(30 0 I ft 0 P Μ vO(30 0 I ft 0 P Μ vO
00< 3 3 XJ 44 Od00 <3 3 XJ 44 Od
pH Ο P I—I 0 · rHpH Ο P I — I 0 · rH
ox -H 0 aj CM 3 M43 P43CM30-ox -H0 aj CM 3 M43 P43CM30-
+J ft U 0 rt P Ή ·Η p rH+ J ft U 0 rt P Ή · Η p rH
• H cd /—I rH I Odl PO ·Η 3·· > P2 P £ Ρ P 44 Od 3 0 0 to od d 0 43 cm P (30 H 0 U 3 CM 3 to 3 0 ·Η 0• H cd / —I rH I Odl PO · Η 3 ··> P2 P £ Ρ P 44 Od 3 0 0 to od d 0 43 cm P (30 H 0 U 3 CM 3 to 3 0 · Η 0
f i i O ©\® *Ή *Hf i i O © \ ® * Ή * H
cdP tX 3 ο ρ xi pcdP tX 3 ο ρ xi p
0 3 · co 0 S O0 3 · co 0 S O
bo e rX ε ε eg ·η d g 10 10 3 ·Ή ΉΛΟ • Η <3 3 ·Η Ρ ·Η Ρ 0 ·Η d3 p S XI <Λ 4b Ρ 003 3 t-HOd rHOd Ο 0 0 ·Η ·3" I 3 .pH.HbO P bo ε w ** < rH 0 Ό, ·Η<ζ0Ο(3θ *bo e rX ε ε eg · η dg 10 10 3 · Ή ΉΛΟ • Η <3 3 · Η Ρ · Η Ρ 0 · Η d3 p S XI <Λ 4b Ρ 003 3 t-HOd rHOd Ο 0 0 · Η · 3 "I 3 .pH.HbO P above ε w ** <rH 0 Ό, · Η <ζ0Ο (3θ *
3X0 4b>x(OxiCMP3X0 4b> x (OxiCMP
Øg ρ ·Η · -©. øoø g cd O 434bpØrH> > g3ft 3 O P( -3 · ·©·Increase ρ · Η · - ©. island cd O 434bpORH>> g3ft 3 O P (-3 · · © ·
33 -rip 3 P 3 X) P33 -rip 3 P 3 X) P
CO 44 ø £ 0 3 O ^ ftCO 44 ø £ 0 3 O ^ ft
rH ·Η O Xt 3 ·Η CMrH · Η O Xt 3 · Η CM
> 3£t0 ØPP· 3> 3 £ t0 EPP · 3
HH ε -Η P 3 -X I-HHH ε -Η P 3 -X I-H
Μ ·ΗΡΡ43.Χ030 0 0 < 3 3 3 rH 4b CM 44Μ · ΗΡΡ43.Χ030 0 0 <3 3 3 rH 4b CM 44
ø ø .H<b003C3PrXø ø .H <b003C3PrX
pH pH 3 g w cd 43 tO ·Η 0 ·Η 0 0 ·ΗpH pH 3 g w cd 43 tO · Η 0 · Η 0 0 · Η
•d xi 0 II II II• d xi 0 II II II
3 3 X3 3 X
•Η ·Η g o• Η · Η g o
4b 4b 3 CJ LO4b 4b 3 CJ LO
P P P ti RP P P ti R
0 0 3 SU I0 0 3 SU I
ft ft iX 3 4b 0 141910ft ft iX 3 4b 0 141910
IIII
Forbindelse IV er værdifuld som antibakterielt middel, profylaktisk tilsætning til foder, terapeutisk middel for fjerkræ, pattedyr og mennesker, og er især værdifuld ved infektiøse sygdomme forårsaget af gram-positive og gram-negative bakterier.Compound IV is valuable as antibacterial agent, prophylactic additive to feed, therapeutic agent for poultry, mammals and humans, and is particularly valuable in infectious diseases caused by gram-positive and gram-negative bacteria.
Forbindelse IV er ved oral indgivelse nyttig til supplerende behandling ved præoperativ sterilisation af tarmen. Både aerob og anaerob flora, som er følsom overfor dette lægemiddel, reduceres i tyktarmen. Efterfulgt af passende mekanisk rensning er det et nyttigt middel ved forberedelsen af tyktarmskirurgi.Compound IV is useful upon oral administration for adjunctive therapy in preoperative bowel sterilization. Both aerobic and anaerobic flora, which are sensitive to this drug, are reduced in the colon. Followed by appropriate mechanical cleansing, it is a useful agent in the preparation of colon surgery.
Forbindelse IV er effektiv ved behandlingen af systemiske bakterielle infektioner hos mennesket ved indgivelse parenteralt i doser i intervallet fra ca. 250 mg til ca. 3000 mg pr. dag i delte doser tre eller fire gange daglig. I almindelighed er forbindelsen effektiv ved indgivelse i en dosis på fra ca. 5,0 til ca. 7,0 mg/kg legemsvægt hver 12 timer.Compound IV is effective in the treatment of systemic bacterial infections in man by administering parenterally at doses ranging from ca. 250 mg to approx. 3000 mg per daily in divided doses three or four times daily. In general, the compound is effective when administered at a dose of from about 5.0 to approx. 7.0 mg / kg body weight every 12 hours.
Fremgangsmåden ifølge opfindelsen illustreres nærmere ved hjælp af følgende eksempler. Eksempel 1-2 illustrerer fremstillingen af udgangsmaterialer, og eksempel 3-4 illustrerer fremgangsmåden ifølge opfindelsen.The process according to the invention is further illustrated by the following examples. Examples 1-2 illustrate the preparation of starting materials and Examples 3-4 illustrate the process of the invention.
Eksempel 1 1. Fremstilling af L-(-)-γ-amino-α-hydroxysmørsyre ud fra ambutyrosln A eller B eller blandinger deraf.Example 1 1. Preparation of L - (-) - γ-amino-α-hydroxybutyric acid from ambiotic acid A or B or mixtures thereof.
Ambutyrosin A (5,0 g) [se beskrivelsen til U.S.A. patentskrift nr. 3.541.078] tilbagesvaledes med 160 ml 0,5N natriumhydroxid i 1 time. Hydrolysatet neutraliseredes med 6N HC1 og kromatograferedes på en kolonne af "CG-50"(NH^+ typen). Den ønskede L-(-)-γ-amino-a-hydroxysmørsyre isoleredes ved fremkaldelse af kolonnen med vand og fjernelse af vandet ved frysetørring. L-(-)-γ-amino-a-hydroxysmørsyre er karakteriseret som et krystallinsk materiale med et smeltepunkt på 212,5-214,5°C [beskrivelsen til U.S.A. patentskrift nr. 3.541.078, kolonne 2, linie 31-38].Ambutyrosine A (5.0 g) [see the description of U.S.A. Patent No. 3,541,078] was refluxed with 160 ml of 0.5N sodium hydroxide for 1 hour. The hydrolyzate was neutralized with 6N HCl and chromatographed on a column of "CG-50" (NH₂ + type). The desired L - (-) - γ-amino-α-hydroxybutyric acid was isolated by evoking the column with water and removing the water by freeze-drying. L - (-) - γ-amino-α-hydroxybutyric acid is characterized as a crystalline material having a melting point of 212.5-214.5 ° C [disclosure to U.S.A. Patent No. 3,541,078, column 2, lines 31-38].
2. Fremstilling af L-(-)-γ-amino-a-hydroxysmørsyre ud fra DL-g-hydroxy-γ-phthalimidosmørsyre A. Dehydroabletylammonium L-a-hydroxy-Y-phthalimidobutyrat;2. Preparation of L - (-) - γ-amino-α-hydroxybutyric acid from DL-g-hydroxy-γ-phthalimidobutyric acid A. Dehydroabletylammonium L-α-hydroxy-Y-phthalimidobutyrate;
Til en opløsning af 25 g (0,1 mol) a-hydraxy-Y-phthalimidosmør-syre (Y. Saito et al., Tetrahedron Letters, 1970, 4863) i 200 ,ml ethanol sattes en opløsning af 29 g (0,1 mol) dehydroabietylamin i 130 ml ethanol. Opløsningen rystedes kraftigt i 1 minut og henstod ved 141910 12 stuetemperatur i 5 timer, i løbet af hvilket tidsrum fine nåle udkrystalliserede. Krystallerne samledes ved filtrering, vaskedes med 50 ml ethanol og lufttørredes til opnåelse af 30,1 g (56%) af en diastereomer af dehydroabietylaminsaltet. Smeltepunkt 93-94°C. [a]^4 = +15° (c. 2,5, MeOH). Rekrystallisation fra 300 ml ethanol gav 23,2 g (43%) af det rene produkt. Smeltepunkt 94-95°C. [a]p4 - +10,8° (c. 2,5, MeOH).To a solution of 25 g (0.1 mole) of α-hydraxy-Y-phthalimido butyric acid (Y. Saito et al., Tetrahedron Letters, 1970, 4863) in 200 ml of ethanol was added a solution of 29 g (0, 1 mol) dehydroabietylamine in 130 ml of ethanol. The solution was shaken vigorously for 1 minute and allowed to stand at room temperature for 5 hours, during which time fine needles crystallized. The crystals were collected by filtration, washed with 50 ml of ethanol and air-dried to give 30.1 g (56%) of a diastereomer of the dehydroabietylamine salt. Melting point 93-94 ° C. [α] 20 = + 15 ° (c. 2.5, MeOH). Recrystallization from 300 ml of ethanol gave 23.2 g (43%) of the pure product. Melting point 94-95 ° C. [α] β 4 - + 10.8 ° (c. 2.5, MeOH).
Yderligere rekrystallisation forandrede ikke smeltepunktet og den specifikke rotation.Further recrystallization did not change the melting point and the specific rotation.
Analyse beregnet for C32H42N2°5/H20: C' 4; H, 8,02; N, 5,07.Analysis calculated for C32H42N2 ° 5 / H2O: C4; H, 8.02; N, 5.07.
Fundet: C, 69,58; H, 8,08; N, 5,07.Found: C, 69.58; H, 8.08; N, 5.07.
B. L-(-)-γ-amino-g-hydroxysmørsyreB. L - (-) - γ-amino-g-hydroxybutyric acid
Til en opløsning af 1,5 g (0,014 mol) natriumcarbonat i 40 ml vand sattes 5,3 g (0,01 mol) dehydroabietylammonium-L-a-hydroxy-γ-phthalimidobutyrat og 60 ml ether. Blandingen rystedes voldsomt, indtil alt faststoffet var blevet opløst. Etherlaget fraskiltes. Den vandige opløsning vaskedes to gange med 20 ml portioner ether og inddampedes til 15 ml under formindsket tryk. Til koncentratet sattes 10 ml koncentreret saltsyre, og blandingen tilbagesvaledes i 10 timer. Efter afkøling fjernedes udskilt phthalsyre ved filtrering. Filtratet inddampedes under formindsket tryk. Remanensen opløstes i 10 ml vand, og opløsningen inddampedes til tørhed. Dette gentoges to gange til fjernelse af overskud af saltsyre. Den sirupagtige remanens opløstes i 10 ml vand og filtreredes til fjernelse af en lille mængde uopløseligt phthalsyre. Filtratet adsorberedes på en kolonne af "lR-12d' (H+, 1 cm x 35 cm), kolonnen vaskedes med 300 ml vand og elueredes med IN ammo-niumhydroxidopløsning. Eluatet opsamledes i 15 ml fraktioner. De nin-hydrin positive fraktioner 10-16 forenedes og inddampedes under formindsket tryk til dannelse af en sirup, som gradvis krystalliserede. Krystallerne tritureredes med ethanol, filtreredes og tørredes i en vakuumeksikator til dannelse af 0,78 g (66%) L-(-)-γ-amino-a-hydroxy-smørsyre. Smeltepunkt 206-207°C. [a]^4 -29° (c. 2,5, H20). Det infrarøde spektrum var identisk med den autentiske prøve, som opnåedes fra ambutyrosin.To a solution of 1.5 g (0.014 mole) of sodium carbonate in 40 ml of water was added 5.3 g (0.01 mole) of dehydroabietylammonium L-α-hydroxy-γ-phthalimidobutyrate and 60 ml of ether. The mixture was shaken vigorously until all the solid had dissolved. The ether layer was separated. The aqueous solution was washed twice with 20 ml portions of ether and evaporated to 15 ml under reduced pressure. To the concentrate was added 10 ml of concentrated hydrochloric acid and the mixture was refluxed for 10 hours. After cooling, separated phthalic acid was removed by filtration. The filtrate was evaporated under reduced pressure. The residue was dissolved in 10 ml of water and the solution evaporated to dryness. This was repeated twice to remove excess hydrochloric acid. The syrupy residue was dissolved in 10 ml of water and filtered to remove a small amount of insoluble phthalic acid. The filtrate was adsorbed on a column of "1R-12d" (H +, 1 cm x 35 cm), the column was washed with 300 ml of water and eluted with 1N ammonium hydroxide solution. The eluate was collected in 15 ml fractions. The ninhydrin positive fractions 10- 16 were combined and evaporated under reduced pressure to form a syrup which gradually crystallized The crystals were triturated with ethanol, filtered and dried in a vacuum desiccator to give 0.78 g (66%) of L - (-) - γ-amino-α -hydroxy-butyric acid Melting point 206-207 ° C [α] 4 -4-29 ° (c. 2.5, H 2 O) The infrared spectrum was identical to the authentic sample obtained from ambutyrosine.
3. Fremstilling af L-(-)-γ-benzyloxycarbonylamino-g-hydroxysmørsyre (VI) L-(-)-γ-amino-a-hydroxysmørsyre (7,4 g, 0,062 mol) sattes til en opløsning af 5,2 g (0,13 mol) natriumhydroxid i 50 ml vand. Til den omrørte blanding sattes dråbevis ved 0-5°C i løbet af et tidsrum på 1/2 time 11,7 g (0,068 mol) carbobenzoxychlorid, og omrøringen fort- 141910 13 sattes yderligere 1 time ved samme temperatur. Reaktionsblandingen vaskedes med 50 ml ether, indstilledes til en pH-værdi på 2 med fortyndet saltsyre og ekstraheredes med fire 80 ml portioner ether. Etherekstrakterne forenedes, vaskedes med en lille mængde mættet natriumchloridopløsning, tørredes med vandfri natriumsulfat og filtreredes. Filtratet inddampedes i vakuum, og deh resulterende remanens krystalliseredes fra benzen til dannelse af 11,6 g (74%) farveløse plader, smeltepunkt 78,5-79,5°C, [a]n * 4,5° (c = 2, CH-ØH). r3. Preparation of L - (-) - γ-benzyloxycarbonylamino-g-hydroxybutyric acid (VI) L - (-) - γ-amino-α-hydroxybutyric acid (7.4 g, 0.062 mol) was added to a solution of 5.2 g (0.13 mol) of sodium hydroxide in 50 ml of water. To the stirred mixture was added dropwise at 0-5 ° C over a period of 1/2 hour 11.7 g (0.068 mol) of carbobenzoxychloride and the stirring was continued for an additional 1 hour at the same temperature. The reaction mixture was washed with 50 ml of ether, adjusted to a pH of 2 with dilute hydrochloric acid and extracted with four 80 ml portions of ether. The ether extracts were combined, washed with a small amount of saturated sodium chloride solution, dried over anhydrous sodium sulfate and filtered. The filtrate was evaporated in vacuo and the resulting residue crystallized from benzene to give 11.6 g (74%) of colorless plates, m.p. 78.5-79.5 ° C, [a] n * 4.5 ° (c = 2 , CH-EH). r
u —I Ju —I J
Infrarødt spektrum [KBr] : y 1740, 1690 caø . Kernexnagnetisk re- C"” O · sonansspektrum (acetone-dg) 6 (i ppm fra TMS): 2,0 (2H,m), 3,29 (2H,d-d, J=6,7 og 12 Hz), 4,16 (IH, d-d, J=4,5 og 8 Hz), 4,99 (2H,s>^ 6,2 (2H, bred) og 7,21 (5H,s).Infrared spectrum [KBr]: y 1740, 1690 caø. Nuclear Magnetic Resonance C o O sonon spectrum (acetone-dg) 6 (in ppm from TMS): 2.0 (2H, m), 3.29 (2H, dd, J = 6.7 and 12 Hz), 4 , 16 (1H, dd, J = 4.5 and 8 Hz), 4.99 (2H, s> 6.2 (2H, broad), and 7.21 (5H, s)).
Analyse beregnet for ci2Hi5N05: C, 56,91? H, 5,97; N, 5,53.Analysis calculated for c12 H15 NO5: C, 56.91? H, 5.97; N, 5.53.
Fundet: C, 56,66; H, 5,97; N, 5,47.Found: C, 56.66; H, 5.97; N, 5.47.
4. Fremstilling af N-hydroxysuccinimldester af L-(-)-γ-benzyloxycar-bonylamino-g-hydroxysmørsyre (VII)4. Preparation of N-hydroxysuccinimide residues of L - (-) - γ-benzyloxycarbonylamino-g-hydroxybutyric acid (VII)
En opløsning af 10,6 g (0,042 mol) af forbindelse VI og 4,8 g (0,042 mol) N-hydroxysuccinimid (G.W. Anderson et al., J. Am. Chem. Soc., j3i6, 1839 (1964)) i 200 ml ethylacetat afkøledes til 0°C, og derpå tilsattes 8,6 g (0,042 mol) dicyclohexylcarbodiimid. Blandingen opbevaredes natten over i et køleskab. Dicyclohexylurinstoffet, som udskiltes, frafiltreredes, og filtratet koncentreredes til ca. 50 ml under formindsket tryk til dannelse af farveløse krystaller af forbindelse VII, hvilke krystaller opsamledes ved filtrering, 6,4 g, smeltepunkt 121-122,5°C. Filtratet inddampedes til tørhed i vakuum, og den krystallinske remanens vaskedes med 20 ml af en benzen-n-hexan blanding til dannelse af en yderligere mængde forbindelse VII.A solution of 10.6 g (0.042 mole) of Compound VI and 4.8 g (0.042 mole) of N-hydroxysuccinimide (GW Anderson et al., J. Am. Chem. Soc., J. 18, 1839 (1964)) Ethyl acetate (200 ml) was cooled to 0 ° C and then 8.6 g (0.042 mole) of dicyclohexylcarbodiimide was added. The mixture was stored overnight in a refrigerator. The dicyclohexylurea, which is excreted, was filtered off and the filtrate was concentrated to ca. 50 ml under reduced pressure to form colorless Compound VII crystals which were collected by filtration, 6.4 g, mp 121-122.5 ° C. The filtrate was evaporated to dryness in vacuo and the crystalline residue was washed with 20 ml of a benzene-n-hexane mixture to give an additional amount of compound VII.
Det totale udbytte var 13,4 g (92%). [a]D = 1,5° (c = 2, CHCl^)· Infrarødt spektrum (KBr) Yc_0 1810, 1755, 1740, 1680 cm 1. NMR (acetone-dg) δ (i ppm fra TMS) 2,0 (2H, m), 2,83 (4H, s), 3,37 (2H, d-d, J=6,5 og 12,5 Hz), 4,56 (IH, m), 4,99 (2H, s), 6,3 (2H, bred), 7,23 (5H,s).The total yield was 13.4 g (92%). [α] D = 1.5 ° (c = 2, CHCl ^) · Infrared spectrum (KBr) Yc_0 1810, 1755, 1740, 1680 cm -1. NMR (acetone-dg) δ (in ppm from TMS) 2.0 (2H, m), 2.83 (4H, s), 3.37 (2H, dd, J = 6.5 and 12.5 Hz), 4.56 (1H, m), 4.99 (2H, s), 6.3 (2H, broad), 7.23 (5H, s).
Analyse beregnet for cigHigN2°7: c' 54,85; H, 5,18; N, 8,00.Analysis calculated for cigHigN2 ° 7: c '54.85; H, 5.18; N, 8.00.
Fundet: C, 54,79, H, 5,21, M, 8,14, 54,70; 5,20; 8,12.Found: C, 54.79; H, 5.21; M, 8.14, 54.70; 5.20; 8.12.
Eksempel 2Example 2
Fremstilling af N-(benzyloxycarbonyloxy)succlnlmld N-hydroxysuccinimid (23 g, 0,2 mol); G.W. Anderson et al., J.Preparation of N- (benzyloxycarbonyloxy) succinic acid N-hydroxysuccinimide (23 g, 0.2 mol); G. W. Anderson et al., J.
Am. Chem. Soc., 86, 1839 (1964), opløstes i en opløsning af 9 g (0,22 141910 14 mol) natriumhydroxid i 200 ml vand. Til den omrørte opløsning sattes dråbevis 34 g (0,2 mol) carbobenzoxychlorid under vandkøling, og derpå omrørtes blandingen ved stuetemperatur natten over til udskillelse af carbobenzoxy derivatet, som samledes ved filtrering, vaskedes med vand og lufttørredés. Udbytte 41,1 g (83%). Rekrystallisation fra benzen-n-hexan (10:1) gav farveløse prismer smeltende ved 78-79°C.Am. Chem. Soc., 86, 1839 (1964), was dissolved in a solution of 9 g (0.22, 14 moles) of sodium hydroxide in 200 ml of water. To the stirred solution was added dropwise 34 g (0.2 mole) of carbobenzoxy chloride under water cooling and then the mixture was stirred at room temperature overnight to separate the carbobenzoxy derivative which was collected by filtration, washed with water and air-dried. Yield 41.1 g (83%). Recrystallization from benzene-n-hexane (10: 1) gave colorless prisms melting at 78-79 ° C.
Eksempel 3Example 3
1. Fremstilling af 61-carbobenzoxykanamycin A1. Preparation of 61-carbobenzoxycanamycin A
En opløsning af 42,5 g (90 mmol) af den frie base af kanamycin A i 450 ml vand og 500 ml dimethylformamid (DMF) afkøledes til under 0°C og omrørtes kraftigt. Til opløsningen sattes dråbevis i løbet af et tidsrum på ca. 2 timer en opløsning af 22,4 g (90 mmol) N-(benzyloxy-carbonyloxy)succinimid i 500 ml DMF. Blandingen omrørtes ved en temperatur på -10°C til 0°C natten over og derpå ved stuetemperatur i 1 dag. Reaktionsblandingen inddampedes under formindsket tryk og ved en temperatur på under 50°C. Den olieagtige remanens opløstes i en blanding af 500 ml vand og 500 ml butanol, filtreredes til fjernelse af uopløseligt materiale og separeredes i to lag. Butanollaget og det vandige lag behandledes henholdsvis med butanol-mættet vand (to portioner af 500 ml) og vand-mættet butanol (to portioner af 500 ml) under anvendelse af en modstrømsfordelingslignende metode. De tre vandige lag forenedes og inddampedes til tørhed under formindsket tryk til dannelse af en olieagtig remanens, hvoraf en del krystalliserede ved henstand ved stuetemperatur. Til remanensen, omfattende krystallerne, sattes ca. 100 ml methanol, som opløste olien og adskilte den fra krystallerne. Efter tilsætning af ca. 300 ml ethanol holdtes blandingen ved stuetemperatur natten over til dannelse af en krystallinsk masse, som samledes ved filtrering. Produktet vejede 44 g. Det indeholdt en lille mængde kanamycin A, som påvistes ved tyndtlagskromatografi under anvendelse af n-propanol-pyridin-eddikesyre-vand (15:10:3:12) som opløsningsmiddelsystem og ninhydrin som reagens.A solution of 42.5 g (90 mmol) of the free base of kanamycin A in 450 ml of water and 500 ml of dimethylformamide (DMF) was cooled to below 0 ° C and stirred vigorously. To the solution was added dropwise over a period of approx. 2 hours a solution of 22.4 g (90 mmol) of N- (benzyloxy-carbonyloxy) succinimide in 500 ml of DMF. The mixture was stirred at a temperature of -10 ° C to 0 ° C overnight and then at room temperature for 1 day. The reaction mixture was evaporated under reduced pressure and at a temperature below 50 ° C. The oily residue was dissolved in a mixture of 500 ml of water and 500 ml of butanol, filtered to remove insoluble material and separated into two layers. The butanol layer and the aqueous layer were treated with butanol-saturated water (two portions of 500 ml) and water-saturated butanol (two portions of 500 ml, respectively) using a countercurrent distribution-like method. The three aqueous layers were combined and evaporated to dryness under reduced pressure to give an oily residue, some of which crystallized on standing at room temperature. To the residue, comprising the crystals, was added ca. 100 ml of methanol which dissolved the oil and separated it from the crystals. After adding approx. 300 ml of ethanol were kept at room temperature overnight to form a crystalline mass which was collected by filtration. The product weighed 44 g. It contained a small amount of kanamycin A which was detected by thin layer chromatography using n-propanol-pyridine-acetic acid water (15: 10: 3: 12) as solvent system and ninhydrin as reagent.
Det rå produkt opløstes i 300 ml vand og kromatograferedes på en kolonne (diameter, 30 mm) af"CG-50"ionbytterharpiks (NH4+ typen, 500 ml). Kolonnen tilførtes en 0,1 N ammoniumhydroxidopløsning, og eluatet opsamledes i 10 ml fraktioner. Det ønskede produkt indeholdtes i glas nr. 10-100, mens kanamycin A blev genvundet fra langsommere bevægende fraktioner og produktets isomere synes at være indeholdt i hurtigere bevægende fraktioner. Fraktionerne 10-100 forenedes og inddampedes til. tørhed under formindsket tryk til dannelse af 24,6 g (45%) af et farveløst produkt 6'-carbobenzoxykanamycin A (II) [6'-Cbz- 141910 15 kanamycin A], som begyndte at smelte og misfarves ved 204°C, og som dekomponerede ved 212°C under gasudvikling, [α]β => +106° (c = 2, H20).The crude product was dissolved in 300 ml of water and chromatographed on a column (diameter, 30 mm) of "CG-50" ion exchange resin (NH4 + type, 500 ml). The column was charged with a 0.1 N ammonium hydroxide solution and the eluate was collected in 10 ml fractions. The desired product was contained in glass # 10-100, while kanamycin A was recovered from slower moving fractions and the product isomers appear to be contained in faster moving fractions. Fractions 10-100 were combined and evaporated. reduced pressure dryness to give 24.6 g (45%) of a colorless product 6'-carbobenzoxycanamycin A (II) [6'-Cbz-kanamycin A] which began to melt and discolour at 204 ° C. and which decomposed at 212 ° C during gas evolution, [α] β => + 106 ° (c = 2, H 2 O).
Tyndtlagskromatografi (silicagel"F254" nin- hydrin) _Rf-værdi_Thin-layer chromatography (silica gel "F254" ninhydrin) _Rf value_
Opløsningsmiddelsystem 61-Cbz-kanamycin A Kanamycin ASolvent system 61-Cbz-kanamycin A Kanamycin A
n-PrOH - pyridin-AcOH-I^O 0,42 0,33 0,15 0,04 (15:10:3:12) (hoved- (andre be- produkt) standdele)n-PrOH - pyridine-AcOH-10 0.42 0.33 0.15 0.04 (15: 10: 3: 12) (main (other product) constituents)
Acetone-AcOH-Η,,Ο 0,24 0,14 (20:6:74) CHCl3-Me0H-c.NH40H-H20 0,76 0,50 (1:4:2:1)Acetone-AcOH-Η, Ο 0.24 0.14 (20: 6: 74) CHCl 3 -MeOH-c.NH40H-H2O 0.76 0.50 (1: 4: 2: 1)
AcOME-n-PrOH-c.NH.OH 0,22 0,04* (45:105:60) x Påvist ved anthron-svovlsyreAcOME-n-PrOH-c.NH.OH 0.22 0.04 * (45: 105: 60) x Detected by anthron sulfuric acid
Slutproduktet viste sig ved tyndtlagskromatografi med et af de afprøvede opløsningsmiddelsystemer at være fulgt af to andre bestanddele. Imidlertid anvendtes slutproduktet uden yderligere rensning til fremstilling af BB-K8 (IV).The final product was found to be followed by two other constituents by thin layer chromatography with one of the solvent systems tested. However, the final product was used without further purification to prepare BB-K8 (IV).
2. Fremstilling af 61-carbobenzoxy-1,3,3"-trl-p-nitro-benzal-kanamycin A (III) 61-carbobenzoxykanamycin A (9,0 g, 14,5 mmol) suspenderedes i 500 ml absolut ethanol ved 24°C og 8,7945 g (58,2 mmol) p-nitrobenzal-dehyd tilsattes. Blandingen opvarmedes til tilbagesvaling. I løbet af dette tidsrum ændrede blandingen sig til en klar, gul opløsning, men ved tilbagesvalingstemperaturen begyndte et hvidt faststof hurtigt at dannes. Blandingen opvarmedes ved tilbagesvaling i 3 timer, køledes så, og produktet opsamledes ved filtrering og vaskedes med en lille mængde absolut ethanol. Udbyttet var 14,0 g (94,5%) af det ønskede produkt III i form af et hvidt, krystallinsk faststof, smeltepunkt 233-234°C (ukorrigeret).2. Preparation of 61-carbobenzoxy-1,3,3 "-trl-p-nitro-benzal-kanamycin A (III) 61-carbobenzoxycanamycin A (9.0 g, 14.5 mmol) was suspended in 500 ml of absolute ethanol at 24-C and 8.7945 g (58.2 mmol) of p-nitrobenzaldehyde were added The mixture was heated to reflux, during which time the mixture changed to a clear yellow solution, but at the reflux temperature a white solid began to rapidly The mixture was heated at reflux for 3 hours, then cooled and the product collected by filtration and washed with a small amount of absolute ethanol, yielding 14.0 g (94.5%) of the desired product III as a white crystalline solid, mp 233-234 ° C (uncorrected).
Analyse beregnet for C47H5iN7°ig: c/ 55,46; H, 5,05; N, 9,63.Analysis calculated for C 47 H 5 N 7 ° g: c / 55.46; H, 5.05; N, 9.63.
Fundet: C, 55,39; H, 5,08; N, 9,60.Found: C, 55.39; H, 5.08; N, 9.60.
3. Fremstilling af 1-[L-(-)-γ-amino-a-hydroxybutyryl]-kanamycin A (IV).3. Preparation of 1- [L - (-) - γ-amino-α-hydroxybutyryl] -canamycin A (IV).
6'-carbobenzoxy-1,3,3"-tri-p-nitrobenzalkanamycin A (5,0 g, 4,91 mmol) opløstes i 50 ml dimethylformamid (DMF) ved 24°C. L-(-)-γ-benzyl- 141910 16 oxycarbonylamino-a-hydroxysmørsyre N-hydroxysuccinimidester (2,064 g, 5,895 mxaol) opløstes i 20 ml DMF ved 24°C og sattes langsomt under voldsom omrøring til opløsningen af Schiff-basen (III) i løbet af 75 minutter. Opløsningen omrørtes ved 24°C natten over. Under anvendelse af dampejektor-vakuum tørredes opløsningen ved lynfordampning ved ca. 40°C. Den lynfordampedes derpå gentagne gange med methanol (100 ml) til dannelse af en viskos olie. Olien opløstes i 100 ml 1:1 dioxan:H20 og 10 ml iseddikesyre tilsattes. Opløsningen anbragtes i en 500 ml Parr kolbe sammen med 2,5 g 5% palladium-på-carbon (Engelhard) og reduceredes under et hydrogentryk på 3,2 kp/cm2 i 4 timer ved 24°C. Det 2 totale trykfald i løbet af dette tidsrum (lukket kolbe) var 6,1 kp/cm , indbefattende periodiske genoprettelser af trykket. Blandingen frafil-treredes igennem en pude af diatoméjord, som derpå vaskedes med tre gange 50 ml 50% vandig dioxan. De forenede filtrater tørredes ved lynfordampning og destilleredes azeotropt med n-butanol (100 ml) og lyn-fordampedes til slut gentagne gange med methanol til dannelse af en viskos olie. Denne opløstes i 30 ml methanol og hældtes langsomt ud i en 1000 ml kold (5-10°C) diethylether under voldsom omrøring. Efter omrøring af den resulterende suspension i 1/2 time i et isbad, samledes udfældningen ved filtrering og tørredes straks i en vakuum-eksikator over P2°5· Pr°duktblandingen vejede 4,9620 g. Tyndtlags-kromatografi viste, at produktet overvejende bestod af forbindelse IV, kanamycin A og nogle spor af di- og trisubstitueret kanamycin A og indbefattede 4-amino-2-hydroxysmørsyre.6'-Carbobenzoxy-1,3,3 "-tri-p-nitrobenzalkanamycin A (5.0 g, 4.91 mmol) was dissolved in 50 ml of dimethylformamide (DMF) at 24 ° C. L - (-) - γ- benzyl oxycarbonylamino-α-hydroxybutyric acid N-hydroxysuccinimide ester (2.064 g, 5.895 mxaol) was dissolved in 20 ml of DMF at 24 ° C and added slowly with vigorous stirring to the solution of the Schiff base (III) over 75 minutes. The mixture was stirred at 24 ° C overnight, using a steam ejector vacuum, the solution was dried by flash evaporation at about 40 ° C. It was then flash-evaporated repeatedly with methanol (100 ml) to give a viscous oil. The oil was dissolved in 100 ml 1: 1 dioxane: H2 O and 10 ml glacial acetic acid were added, the solution was placed in a 500 ml Parr flask with 2.5 g of 5% palladium-on-carbon (Engelhard) and reduced under a hydrogen pressure of 3.2 kp / cm 2 for 4 hours at The 2 total pressure drop during this time (closed flask) was 6.1 kp / cm, including periodic pressure recoveries. n pad of diatomaceous earth which is then washed with three times 50 ml of 50% aqueous dioxane. The combined filtrates were dried by flash evaporation and azeotroped with n-butanol (100 ml) and finally evaporated repeatedly with methanol to give a viscous oil. This was dissolved in 30 ml of methanol and poured slowly into a 1000 ml cold (5-10 ° C) diethyl ether with vigorous stirring. After stirring the resulting suspension for 1/2 hour in an ice bath, the precipitate was collected by filtration and immediately dried in a vacuum desiccator over P2 ° 5 · The product mixture weighed 4.9620 g. Thin layer chromatography showed that the product consisted predominantly of compound IV, kanamycin A and some traces of di- and trisubstituted kanamycin A and included 4-amino-2-hydroxybutyric acid.
De forskellige fraktioner separeredes under anvendelse af en 40 x 100 mm glaskolonne pakket med"Amberlité1 ® IRC-40 (NH^+ form, type I, 1.00/200 mesh) til tilvejebringelse af et harpiksleje på ca. 980 mm. Kolonnen fødedes med 4,600 g af den rå blanding opløst i 15 ml vand. Kolonnen elueredes med vand med en NH^OH gradient på fra 0 til 2 N, idet eluatet opsamledes i 15 ml fraktioner. 295 fraktioner opsamledes, og derpå vaskedes kolonne med 1,5 liter 3N NH^OH. Fraktionerne samledes på basis af optisk rotation, og faststofferne isoleredes derfra ved lyofilisering.The various fractions were separated using a 40 x 100 mm glass column packed with "Amberlité1® IRC-40 (NH 2+ form, type I, 1.00 / 200 mesh) to provide a resin bed of about 980 mm. The column was fed 4,600 The column was eluted with water with an NH 2 OH gradient of 0 to 2 N, the eluate collected in 15 ml fractions, 295 fractions collected and then the column washed with 1.5 liters of 3N The fractions were collected on the basis of optical rotation and the solids were isolated therefrom by lyophilization.
Korrektion til vægt af Vægt samlet rå- BiologiskCorrection to Weight of Weight Total Raw Biological
Fraktion nr. S ammens ætn ing (g) produkt (g) styrke Udbytte, % 120-150 Kanamycin A 1,1654 1,257 809 42,6Fraction No. S amine etching (g) Product (g) Strength Yield,% 120-150 Kanamycin A 1,1654 1,257 809 42.6
228-241 BB-K8 0,6523. 0,7036 786±101* 19,2KX228-241 BB-K8 0.6523. 0.7036 786 ± 101 * 19.2KX
x Gennemsnit af 4 plade forsøg, + standardafvigelse, xx Hvis der tages hensyn til genvundet kanamycin A, var omdannelsen til BB-K8 (IV) 33,5%.x Average of 4 plate trials, + standard deviation, xx If recycled kanamycin A is taken into account, conversion to BB-K8 (IV) was 33.5%.
17 U191017 U1910
Ved dette forsøg var udbytteforholdet BB-K8/kanamycin A = 0,45.In this experiment, the yield ratio was BB-K8 / kanamycin A = 0.45.
Den ved fremgangsmåden ifølge opfindelsen udvundne forbindelse IV er identisk med det autentiske produkt, som opnåedes ved den i beskrivelsen til dansk patentansøgning nr. 3461/72 omhandlede fremgangsmåde. Smeltepunkt 194°C (dekomponering) . Ια]^ = +85° (c=2, E^O) . Relativ styrke imod B. subtilis (agarplade) = 960^,ug/mg (standard: fri base af kanamycin A).The compound IV obtained by the process according to the invention is identical to the authentic product obtained by the method described in the description of Danish Patent Application No. 3461/72. Melting point 194 ° C (decomposition). [Α] D = + 85 ° (c = 2, E + O). Relative potency against B. subtilis (agar plate) = 960 µg / mg (standard: free base of kanamycin A).
Analyse beregnet for ^22^43^0^3,2^0()3: c, 40,62; H, 6,68; N, 9,87.Analysis calculated for ^ 22 ^ 43 ^ 0 ^ 3.2 ^ 0 () 3: c, 40.62; H, 6.68; N, 9.87.
Fundet: C, 40,21; H, 6,96; N, 9,37, 39,79; 6,87; 9,49.Found: C, 40.21; H, 6.96; N, 9.37, 39.79; 6.87; 9.49.
Eksempel 4Example 4
Fremstilling af disulfatsaltet af 1-[L-(-)-γ-amino-a-hydroxybutyryl]-kanamycin A (BB-K8, 2H2SO4) 35 g 1-[L-(-)-γ-amino-a-hydroxybutyryl]kanamycin A i form af mono-hydrogencarbonattrihydrat opløstes i 125 ml deioniseret vand. Opløsningen havde en pH-værdi på ca- 9,0. pH-værdien sænkedes til 7 - 7,5 med 50 volumenprocent svovlsyre.Preparation of the disulfate salt of 1- [L - (-) - γ-amino-α-hydroxybutyryl] -canamycin A (BB-K8, 2H2SO4) 35 g of 1- [L - (-) - γ-amino-α-hydroxybutyryl] kanamycin A in the form of mono-hydrogen carbonate trihydrate was dissolved in 125 ml of deionized water. The solution had a pH of about 9.0. The pH was lowered to 7 - 7.5 with 50% by volume sulfuric acid.
8 1/2 g aktivt kul tilsattes, og blandingen opslæmmedes ved stuetemperatur i 1/2 time. Carbonet fjernedes ved hjælp af filtrering og vaskedes med 40 ml vand. Vaskevandet sattes til filtratet.8 1/2 g of activated carbon was added and the mixture was slurried at room temperature for 1/2 hour. The carbon was removed by filtration and washed with 40 ml of water. The wash water was added to the filtrate.
pH-værdien for det ovennævnte forenede filtrat-vaskevand indstilledes til 2-2,6 med 50 volumenprocent svovlsyre. En stor mængde carbondioxid udvikledes. Opløsningen fik lov at stå mider vakuum og omrøring i 20 minutter til uddrivelse af yderligere carbondioxid.The pH of the above combined filtrate wash water was adjusted to 2-2.6 with 50% by volume sulfuric acid. A large amount of carbon dioxide developed. The solution was allowed to stand under vacuum and stirring for 20 minutes to evaporate additional carbon dioxide.
8 1/2 g aktivt kul sattes til den afgassede opløsning. Blandingen opslæmmedes i 1/2 time ved stuetemperatur. Carbonet fjernedes ved filtrering og vaskedes med 35 ml deioniseret vand. Vandet sattes til filtratet.8 1/2 g of activated carbon was added to the degassed solution. The mixture was slurried for 1/2 hour at room temperature. The carbon was removed by filtration and washed with 35 ml of deionized water. The water was added to the filtrate.
pH-værdien for det forenede filtrat-vaskevand indstilledes til 1-1,3 med 50 volumenprocent svovlsyre. Denne opløsning sattes under hurtig omrøring i løbet af et tidsrum på 10 minutter til 600 - 800 ml methanol (3-4 volumener methanol). Blandingen omrørtes i 5 minutter ved pH 1 - 1,3, ledtes igennem en 100 mesh sigte , omrørtes i 2 minutter og fik lov at sætte sig i 5 minutter. Det meste af den ovenpå flydende væske dekanteredes fra. Den resterende opslæmning filtreredes, vaskedes med 200 ml methanol og vakuumtørredes ved 50°C i 24 timer. Udbyttet af amorft BB-K8, 2H2S04 var 32 - 34 g. [a]£2 = +74,75° (H20). Dekomponering ved 220-223°C.The pH of the combined filtrate wash water was adjusted to 1-1.3 with 50% by volume sulfuric acid. This solution was added with rapid stirring over a period of 10 minutes to 600 - 800 ml of methanol (3-4 volumes of methanol). The mixture was stirred for 5 minutes at pH 1 - 1.3, passed through a 100 mesh sieve, stirred for 2 minutes and allowed to sit for 5 minutes. Most of the liquid upstairs was decanted. The remaining slurry was filtered, washed with 200 ml of methanol and vacuum dried at 50 ° C for 24 hours. The yield of amorphous BB-K8, 2H2SO4 was 32 - 34 g. [Α] 25 D = + 74.75 ° (H 2 O). Decomposition at 220-223 ° C.
Claims (1)
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US33037773A | 1973-02-07 | 1973-02-07 | |
US33037773 | 1973-02-07 |
Publications (2)
Publication Number | Publication Date |
---|---|
DK141910B true DK141910B (en) | 1980-07-14 |
DK141910C DK141910C (en) | 1980-11-24 |
Family
ID=23289488
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
DK59374AA DK141910B (en) | 1973-02-07 | 1974-02-04 | Process for the preparation of 1- (L - (-) - gamma-amino-alpha-hydroxybutyryl) -canamycin A or acid addition salts thereof. |
Country Status (10)
Country | Link |
---|---|
JP (1) | JPS575797B2 (en) |
CA (1) | CA1021329A (en) |
CH (1) | CH593994A5 (en) |
DK (1) | DK141910B (en) |
FI (1) | FI58134C (en) |
HU (1) | HU170378B (en) |
NL (1) | NL182962C (en) |
SE (1) | SE417971B (en) |
SU (1) | SU667142A3 (en) |
YU (1) | YU30674A (en) |
Families Citing this family (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPS5342038B2 (en) * | 1973-11-19 | 1978-11-08 | ||
JPS5910360B2 (en) * | 1975-01-24 | 1984-03-08 | 萬有製薬株式会社 | Improved production method of 1-N-[L-(1)-γ-amino-α-hydroxybutyryl]-kanamycin derivative |
IT1139844B (en) * | 1981-11-25 | 1986-09-24 | Corvi Mora Paolo | PROCESS FOR THE PREPARATION OF AMIKACINA |
-
1974
- 1974-01-29 JP JP1152474A patent/JPS575797B2/ja not_active Expired
- 1974-02-04 DK DK59374AA patent/DK141910B/en not_active IP Right Cessation
- 1974-02-04 NL NLAANVRAGE7401517,A patent/NL182962C/en not_active IP Right Cessation
- 1974-02-05 FI FI309/74A patent/FI58134C/en active
- 1974-02-06 HU HUBI488A patent/HU170378B/hu unknown
- 1974-02-06 CA CA191,868A patent/CA1021329A/en not_active Expired
- 1974-02-06 SU SU741992760A patent/SU667142A3/en active
- 1974-02-07 CH CH171774A patent/CH593994A5/en not_active IP Right Cessation
- 1974-02-07 YU YU00306/74A patent/YU30674A/en unknown
- 1974-02-07 SE SE7401669A patent/SE417971B/en not_active IP Right Cessation
Also Published As
Publication number | Publication date |
---|---|
SU667142A3 (en) | 1979-06-05 |
NL182962B (en) | 1988-01-18 |
DK141910C (en) | 1980-11-24 |
YU30674A (en) | 1982-02-28 |
HU170378B (en) | 1977-06-28 |
FI58134B (en) | 1980-08-29 |
CA1021329A (en) | 1977-11-22 |
SE417971B (en) | 1981-04-27 |
JPS49102644A (en) | 1974-09-27 |
NL182962C (en) | 1988-06-16 |
NL7401517A (en) | 1974-08-09 |
JPS575797B2 (en) | 1982-02-01 |
CH593994A5 (en) | 1977-12-30 |
FI58134C (en) | 1980-12-10 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
DE2742949C2 (en) | 4-N-acylfortimicin B derivatives and their uses | |
EP0041355A1 (en) | Novel erythromycin compounds | |
JP2010527913A (en) | Antibacterial 1,4,5-substituted aminoglycoside analogues | |
CA2082216A1 (en) | Polycyclic conjugates | |
HU177261B (en) | Process for preparing new aminoglycoside derivatives | |
US3808198A (en) | Lividomycin b derivatives | |
US4170642A (en) | Derivatives of kanamycin A | |
NO139483B (en) | ANALOGICAL PROCEDURE FOR PREPARING NEW PSEUDOTRISACCHARIDES | |
EP0009670A1 (en) | Aminoglycoside derivatives, process for their preparation and pharmaceutical compositions | |
NO144851B (en) | ANALOGY PROCEDURE FOR THE PREPARATION OF THERAPEUTICALLY ACTIVE KANAMYSIN A OR KANAMYCIN B DERIVATIVES | |
US4337248A (en) | Paromomycin containing compounds and method of use | |
DK141910B (en) | Process for the preparation of 1- (L - (-) - gamma-amino-alpha-hydroxybutyryl) -canamycin A or acid addition salts thereof. | |
CS228516B2 (en) | Method for the production of guanyled aminoglykosides | |
US4181797A (en) | 1-N-(ω-amino-α-hydroxyalkanoyl) derivatives of 4'-deoxy-6'-N-methylkanamycin A | |
CA1046058A (en) | Preparation of a novel aminoglycosidic derivative and its salts thereof | |
EP0040764B1 (en) | Novel aminoglycosides, and antibiotic use thereof | |
US3974137A (en) | Process for the preparation of 1-[L-(-)-γ-amino-α-hydroxybutyryl]-kanamycin A (RD-1341A) | |
US4362866A (en) | Aprosamine derivatives | |
GB2086383A (en) | 5-deoxyapramycin | |
US4205070A (en) | 6'N-Alkyl- and 6',6'-di-N-alkyl derivatives of fortimicins A and B | |
EP0072351A1 (en) | Aminoglycoside derivatives, processes for their production, pharmaceutical compositions containing them and such derivatives for use as pharmaceuticals | |
FI56388C (en) | REFERENCE TO A FRAMEWORK FOR THERAPEUTIC USE OF THERAPEUTIC CHARACTERISTICS OF 1-N-NY-AMINO-ALPHA-HYDROXIBUTYRYL SUBSTITUTES FOR CANANYCIN (A ELLER B) -FOERENING | |
DK142991B (en) | PROCEDURE FOR THE PREPARATION OF 1- (L - (-) - Gamma-AMINO-ALFA-HYDROXY-BUTYRYL) -CANAMYCINE A OR ACID ADDITION SALTS. | |
US4016158A (en) | Cephalosporin derivatives | |
FR2465746A1 (en) | 1-EPI-2-DESOXYFORTIMICIN B AND ITS DERIVATIVES USEFUL AS ANTIBIOTICS |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PUP | Patent expired |