FI60217C - FRAME FOR THE FRAMSTATION OF AV 1- (L - (-) - NY-AMINO-ALPHA-HYDROXIBUTYRYL) CANANYCIN A - Google Patents

Description

Γβΐ ANNOUNCEMENT, n 0. n W (11) UTLÄGGNINGSSICIUFT 6021 7 (^) Patent cieddelat ^ '* ~ 0' (51) Ky.ik.Wci.3 C 07 H 15/22 ENGLISH I — FI N LAN D (21) Petent application * - Pttenttrwdknlng 3270 / 71 + (22) Hakemlipttv * - An * 6knlnf * di | 11.11.71+ (23) Alkupihrl —Glltlghetadig II.II.7I + (41) Public interpreters - Bllvlt offentllg 15.05.75

National Board of Patents and Registration ..A ......,. ,. , _,, (44) Date of Nihtlvlkslpenon and hearing | - n n

Patent-och registrstyrelsen 'Antttkan utlagd oeh utl.skrlften publicerad 31.08.81 (32) (33) (31) Privilege claimed— Begird priority 11 + .11.73 23.05.7 ^ USA (US) I + I5589, 1 + 72780 (71) ) Bristol- Company, 3 + 5 Park Avenue, New York, NY, USA (72) Takayuki Naito, Tokyo, Susumu Nakagawa, Tokyo, Masahisa Oka, Tokyo,

Japan i-Japan (JP) (7I +) Oy Kolster Ab

(5I +) Process for the preparation of 1- / L - (-) - Y'-amino-Q1-hydroxybutyryl-7-kanamycin A - Preparation of 1- (L) (-) - Y'-amino-β-hydroxybutyryl-7-kanamycin A A

The present invention relates to a process for the preparation of 1-N, N - (-) - (N) -amino (-hydroxybutyryl) 7-kanamycin A of the formula CH0-NH0 HO-2

/B

HO I NH0 0 CH OH HO —-7v ^ \

HO- ^ * / ^ V-NH

\ - 0 / C = 0

H2N / H0-CH

»° \ / H

7¼ HH2 2 60217 which method is characterized by the formula

H S

ho _ ch2-n-c-o-ch2-c6h5

HO

/ III

A compound according to H0 z ° - HO-i - Λ “VL o / ^ - zz / ^ Λ / H0 \ / 0 /, where Z is a group H V_ Ά. / ~ \ HJ V- -N = C / NO2 ' -N = C \ / '_N = C \ / 0CH3 »

HO

hV \ H

-N = C 'V 7 or -N = C-C ~ -CH 3 ^ ch 3 is reacted in situ in a molar ratio of at least 1: 0.5 with a compound of formula 0 2 2

) N-O-C-CH-CH 0 -CH-NH-C-O-CH 0 -C (H). XX

5 "5 and the reaction product is hydrogenated in situ to give the compound of formula IV.

3,60217

Kanamycin A in the formula 6 'V CH2-NH2 H0 - H0 _ ° \

,, NH

, „CH20H HO-1 — _T \ H0 —iL 5 Ας ^ Α1

VaJ 5 "/ m2 H? N - ^ ° \ / 2 ^ T7 ^" / H0 Ö has four primary amine functions at the 1-, 3-, 6'- and 3 "positions of the molecule.

It has been shown that the 6'-amine function is the most reactive and the 1-amine function is the second most reactive when treated with an electrophilic substance. Both the 3 'and 3' positions are less reactive than either the 1- or 6'-amine function, but they react to give a low amount of undesired acylated substances.

The preparation of 1- [L - (-) * γ-amino- (C-hydroxyhutyryl) 7-kanamycin A (BB-K8) is known from FI patent publication 56 388. However, according to the method described in this publication, the yield of BB-K8 is rather low. . In Example 2, 552 mg (19.2%) of BB-K8 were obtained. However, the product contained only 560 μg / mg BB-Κδ (purity 56%). Purification of the product gave 98 mg of pure 96- $ product, corresponding to a total yield of 3.85% of 96 # BB-K8. By the process of the present invention, BB-K8 is obtained, as can be seen from the examples below, in significantly better yields.

The preparation of a compound of formula III is described in F1 patent application No. 309/7 **.

The preparation of a compound of formula IV using aldehydes of formula III as an intermediate is described in FI patent application 309/7 * +. However, it has reacted a compound of formula III with L- (-) - "N-Bentyl] oxycarbonylamino (N-hydroxybutyric acid N-hydroxysuccinimide ester) using LY-benzyloxycarbonylamino-.beta.-hydroxybutylimido N-hydroxy-5-norbornene-2,3-dicarboxyimide ester (XX).

and 60217

When a compound of formula XX is used, the selectivity of the reaction is better than when a succinimide ester is used. In Example 1 of patent application 309/7 * +, the yield of the final product BB-K8 is 19 »* +% or calculated on the basis of reacted kanamycin A 33.5% · The yield of BB-K8 using the norbornene derivative of formula XX is as follows: from Example U can be seen, 36.52%. At the same time, the number of by-products is smaller.

The reaction between a compound of formula III and a compound of formula XX is preferably carried out in a molar ratio of about 1: 1 at a temperature in the range of about -10 ° to about + 35 ° C, more preferably + 5 ° to about 25 ° C, with a reaction time of at least 1-3 hours. after which the organic solvent is removed before hydrogenation. The in situ hydrogenation is carried out in the presence of a metal catalyst such as palladium, platinum, Raney nickel, rhodium, ruthenium and nickel, but especially in the presence of a palladium catalyst and in particular a palladium on carbon catalyst, in water or in a water-miscible solvent system. solvent. These include water and dioxane, tetrahydrofuran, ethylene glycol dimethyl ether, propylene glycol dimethyl ether and the like. The pH of the reaction mixture is 3 to 5, but most preferably h.

The process of the invention is preferably carried out by treating a compound of formula III in situ or after isolation with a compound of formula XX in a molar ratio: about 0.5 to 1.0 moles of compound XX per mole of compound III, in the temperature range + 5-35 ° C for one hour to 3 hours in absolute ethanol, methanol, n-propanol, sec-butanol, tert-butanol, methylene chloride, tetrahydrofuran, dioxane, dimethylformamide or acetone, or a mixture thereof with water; the organic solvent is then removed and the residue is hydrogenated in situ with water in the presence of a metal catalyst such as palladium, platinum, Raney nickel, rhodium, ruthenium and nickel, but especially palladium or especially palladium on carbon, preferably in a mixture of water and water-miscible solvent, of the systems: water and dioxane, tetrahydrofuran, ethylene glycol dimethyl ether and propylene glycol dimethyl ether, but most preferably water, at pH 3-5, more preferably * + to give a compound of formula IV.

The process according to the invention can also be carried out by carrying out several reaction steps in situ, in which case the compound of the formula IV is obtained in the same very good yields at a lower cost.

For example, a compound of formula XX may be prepared in situ and reacted with a compound of formula IIT in a single reaction step.

5,60217

Thus, a compound of formula TV can be prepared by reacting a compound of formula III with formulas 0 0

H-O-C-CH-CH 0 -CH-NH-C-O-CH-CH 2 H 2 -V

, £ - 2 2 o 5

OH

and 0

} N-OH XIX

O

in the presence of dicyclohexylcarbodiimide.

In this case, about 0.5 to 1.0 moles of both compound VI and dicyclohexylcarbodiimide and about 0.005 to 0.25 moles of compound XIX are used per mole of compound III, the reaction temperature is about -10 to + 35 ° C, more preferably +5 to about 25 ° C, the reaction time is at least 10 hours, and one of the systems described above is used as the solvent. The organic solvent is then removed and the residue is hydrogenated in situ as described above to give a compound of formula IV.

The compound of formula IV is a valuable antibacterial agent, a food supplement in animal nutrition, a therapeutic agent in poultry and animals, including humans, and is particularly valuable in the treatment of infectious diseases caused by gram-positive and gram-negative bacteria.

The antibacterial and therapeutic properties of the compound of formula IV are discussed in FI patent publication 56,388.

The following examples illustrate the invention.

Example 1 1-N-ZL - (-) - β-amino-N-hydroxybutyryl-kanamycin A (BB-K8, IV) via benzaldehyde-Schiff base

To a stirred solution of 6'-Cbz-kanamycin (II) (6.19 g, 0.01 mol) in 50 ml of 50%. THF was added 3 mL (0.03 mol) of benzaldehyde. The mixture was stirred at room temperature for 3 hours, cooled to 5 ° C and treated with a solution of compound XX (1+ g, 0.01 mol) in 50 ml of THF.

The mixture was stirred for 1 hour at 5 ° C, the pH was adjusted to 2 with 6N HCl, stirred for a further hour at room temperature, then the organic solution was evaporated in vacuo. The resulting aqueous solution was washed with 20 mL of CH 2 Cl 2, the pH was adjusted to 6,602,17 1+ with 10% NH 4 OH, and the solution was hydrogenated overnight at normal pressure at room temperature using 1.25 g of 10% palladium on carbon as a catalyst. The catalyst was filtered off and washed with 20 ml of water. The filtrate and washings were combined, the pH was adjusted to 8 with $ 10 NH 4 OH and the solution was passed through a CG-50 column (NH 4, 200 mL), eluting with 0.1 L of water, 1.2 L of 0.1 -N NH 4 OH, with 2.06 liters of 0.3-n NH 4 + OH, 0.92 liters of 0.5 N NH 4 OH and finally with 1.2 liters of 1.0 N NH ^ 0H added. The eluate was collected in 20 ml fractions and divided into subsequent fractions based on Rf value (TLC, silica gel plate S-110, ninhydrin) and a disk test using B. subtilis PCI 219 and P. aerugenosa A 98-3. Each fraction was evaporated in vacuo and lyophilized.

7 60217 i id • h: d t> p d: d P d p · Η

□ Or- co Ό p -P

r- · Η d P -P

d W id ® -P dd IW λ P · Η <L> P id CP t> a co dpd -π CP PP id ω dd CP d: d id · η co · η + -PI — IE · co d * H + · Η H d ® • h co On On t— OL * idid d CO >> CM CM CO T- H!> P d • he >> ω id: dd ddWWWWto H-pid * h O id t III 0Ö dpa id

ρ <33ρρρρρΡΡΡ · η ®> 3 P

dPAJPPPPPPCPTJ M id d p d ro a! <L> ή d cm mm co d

co d P · Η P

13¾ d o H

r- · 13¾ P Αί H P

r- O d <L), r ~ 3 cn mm d d ο

P CM AI H P P

O I H, d H

O νο o P d d H

ο σν o \ in daw • η -p · - m -π d co CP · - 1- d p d

0 co -h W

g -π p I

3 ο ο CP

0) p o PP

d -π ft d p id 'M-3 · Η, ¾

^ Vt ^ dp AJ

Vi. Vt Vi Vt H- ~ -d d o m o h d d o pm co vo m cm cm p h ^ p d -— - · - ή h d d ω o d d

dbD bp öO M bO bp ppp O

ws a a a a a ρ p p;

O f- (· H * H CO

dm m vo o vo «Ai o Ai • ho co t— c— o - = t · · η B od vo p-mco <- * - · η d H" -3 CL, t- t— d Ρ · Η p Pi do d 1 P <A! J d co ΰ odpd · η ή A · dd HHPP ® h -H d Ö d Ai

.. p -d P ω I

K P O p A! O ·

O 3 3 Ai I 00 C

P · CP CO Ah On VO P

«> 5 r- O p S co d p; cm cm pd · Η iO, p • d ddddd hpl> »m <3 pi III i IH pr> ο ω • nm mm in in ο · ηοο Ph p P ^ Ci ΛΛΛΛΛ OO -P CÖÖ • HO OOOO i- d. ri CO CO · Η to CP p W pd On

p I · η H O CM

> id p · · η a

d p d d p 3 W

d I h d H p P I

p m id p d d cp

• ri ·> B Id O CO Pi CP

Ο Ο P d Ai d P id p · · d PP id · Η CP fed · Ρ cd · η> a aj p · η ddd • ri ddppp »pp id id« - p Pi pp · - P d P dpp Ο · Η d Pi S >> id

·· · η d C -'d I

d P 3 · Η Ah X PP

P P P PP

d · - o \ t— m t— p —-d d d P Ov VO VO CT \ O in p ό p p "• H ON '-'-'- CMCM -Hdo P P d

Ail lllll d co d p p-ri PO cm cm o co co O co co p P d 3 On LA VO f— O '. m 3 · Η Ή · Η · Η (li i-i-i-i-CM copid d dp

P P O O CO., LTD

p ä p d d b O d P co co s A! CO P d o o> 3 p p d • rt rH id O O d O CP O d W W Ah

• H

^ oj cn ~ = t lt \ vo c \ j is u 8 60217

Fraction U (1.877 g) was dissolved in 10 ml of deionized water. The pH of the solution was adjusted to 6 with 10 # HCl and the solution was adsorbed onto a CG-50 column (NH 4, 15 mL) which was eluted with 100 mL of 0.3 N NH 4 OH and 200 mL of 0.5 N HCl. NH ^ OHtta. The eluate was collected in 20 ml fractions. Tubes 6 and 8-17 were ninhydrin positive. The contents of tubes 8-17 were combined and concentrated in vacuo to about 1 mL. The concentrate was treated with 3 mL of MeOH and filtered. The filter was washed with 0.5 mL of $ 75 aqueous MeOH. The filtrate and washings were combined, diluted with an additional 10 mL of MeOH and left at room temperature overnight. The amorphous precipitate was filtered, and crystalline embryos were seeded into the filtrate to give compound IV as colorless crystals, which was identical to the authentic sample. The filtrate was evaporated in vacuo and lyophilized. The yield and experiment of each fraction are shown in the following table.

IV. Quantity Test Yield B. subtilis K. pneumoniae B. subtilis K. pneumoniae

Amorphous 70 mg 737 [mu] l / mg 927 [mu] l / mg 0.9% 1.1% powder

Crystals 1098 mg 1163 "1087" 19 Ϊ 19 Ϊ

Lyophilized 180 mg 608 "50U" 1.9 ^ 1.6% powder BB-K 11 is 3 "-N- / L - (-) -" γ-amino-cis-hydroxybutyrylZ-kanamycin A.

BB-K 29 is 3-N- [(-) - iT-amino-? N-hydroxybutyryl] -kanamycin A.

The starting material was prepared as follows: Preparation of N-hydroxy-5-norbornene-2,3-dicarboximide ester (XX) of L-1-benzyloxycarbonylamino-β-hydroxybutyric acid Dicyclohexylcarbodiimide (DDC) (2.06 g, 0.01 mol) was added at 10 ° To a stirred solution of L-γ-benzyloxycarbonylamino-β-hydroxybutyric acid (2.53 g> 0.01 mol) and compound XIX (1.79 g, 0.01 mol) in 50 ml of dry THF (tetrahydrofuran). The mixture was stirred overnight at room temperature. The precipitated dicyclohexylurea was removed by filtration and washed with THF.

The filtrate and washings were combined and evaporated in vacuo. The oily residue (U, 5 g) was passed through a column of silica gel and eluted with ethyl acetate to give U.0 g (96%) of the active ester XX as a colorless oil. Infrared spectrum (IR, Neat): 3300, 1820, 1780, 1735, 1720, 1695 cm-1. Nuclear Magnetic Resonance Spectrum (NMR) (CDCl 3 + 1 drop of D 2 O): δ (ppm) 1.63 (2H, q, J = 9Hz), 2.15 (2H, m) 3, U0 (6h, m), U .60 (1H, dd, J = 7 and 5 Hz), 5.13 (2H, s), 6.20 (2H, broad), 7.1 * (5H, s).

7 60217 I: 3 • H * 3 <0> O β: 3 P 3 ρ · η ao «- to t) -p 4-3 T- · η q <υ p 3« "ö 3 -μ c 3 i Ai, ρ p -h <u P: 3 pq i >> 0 β -p 3 · η pq W tee to oi β PQ 3: 3: 3 · Η to -rl + 4-3 Η Θ to β · Η + · Γ4 HC 0

• H 0 OM Om O As β: 3 H

0 A> cm cm oo * - ha φ β • HS >> (U: tö: cfl β 3 3 Ai Ai Ai Ai 0 rl 43: qJ · Η OCIIII CÖ β PS = 3 4-3 co 3 pq pq PQ pq · Η 3 As 43 G +3 pq pq pq pq Ό to: 3 β 43

3 not M 0 -H

3 cm co co to β en 3 0 · η 0

tÖ O 1 — I

r- '-¾¾ %% 43 Jtj lH β T- o β 0 Ό ro co m cö tö ai 0 CM M H 43 tl)

O I H rC H

X O MO O 033 H

O On om ctm 3 S 0 • H pr 1— to * h to ao pq r- T- G -P β O to · Η Ai

S -h 43 I

3oo PQ

0 -P o PQ

a -h ft 1), ο «d r- '· ^' 1-3 · Η Ai -« r- »^ 3 43 Ai

Vä. Vä. H β G

O (O O rH 3 3 0

p CO 00 MO CO CM CM 0H P

β —- --- ^ · Η H cd 3 0 O 3 β

3 M ÖDMMbDM p Ai P O

«E eeaee 0 φ p O H · Η · Η 3 difference CO MO e— O MO Ai Ai O Ai

• HO 00 E— t— O pj- · Η S O

3 MO _d-rOOOr - <- Ή β I — I • '-i PG r- .— 3 0 Ή 4> ft 3 0 3 1 P, X · Cd

to β O 3 P

3 -H · Η x 3 g H H P -P ® rH -H 3 G β Ai

p Ό 0 3 I

Ä p O 0 M o · O 3 3 Ai I CO β ps- 0 0 Ai OM MO 0 K A »<- 0 o a 0 3 pi CM CM p

3 -H: θ, P

• β βββββ HP As H <3 PI CII t I rH PAO 0 • H to CO CO UM LCM O · ΗΟΟ pH 0 pL, * · «· '»> *> »> OOP 3β • HO OOOO * - 3 -HW 0 "rl w pq p (¾ · ρ β θ \

0 l -H 1 — I O CM

a s3 p · -h a β P G 3 p 3 fcci

3 I H 3 H P 0 I

P (O: 3 rH 3 β PQ

• H «· S i3 O 0 ft W

O O 0 β Ai 3 H: 3 0 · · 3 rH Φ: 3 · Η PQ Ai · Ό W · <~ 3 o A! .

rH · <Η β β β • H 3 β Φ 0 ..

i> ρ 0: 3 t3 T- P Ρβ p p «-

Ρ β 0 3 P P

O · γ4 β Ρβ .H Ai Ai

• H β β: 3: 3 I

β P 3 -H Ai Ai «

P P P PQ

G t-0 \ t— COE— 0 ^ 3 3 3 0 OM MOMOOmOLCM p p P P "• H OM r- t— 1 CM CM · Η 3 O Ρ ρβ

Ail lllll β 0 3 Φ 0 · Η PO CM CM Ο ΟΟ CO O00 Ρ Ρβ

3 OM U ~ \ MO t— O. to 3 · Η -H Ή -H

pit r-i— t— CM 00: 3 β β · H

ρ ρ Ο Ο 0 0 Ρ Ρ 3 3 As

0 3 0 0 0 E

Ai 0 Ρ 3 O O S 0 0 β • Η Η «3 Ο 0 3 O pq Ο β Ai Ai Ai

• M

CM CO -tf to, MO r- CM CO

β & ·, 8 60217

Fraction Π (1.877 g) was dissolved in 10 ml of deionized water. The pH of the solution was adjusted to 6 with 10 # HCl and the solution was adsorbed onto a CG-50 column (NH 4, 15 mL) eluting with 100 mL of 0.3 N NH 4 OH and 200 mL of 0.5 ~ n NH ^ OH ^ a. The eluate was collected in 20 ml fractions. Tubes 6 and 8-1T were ninhydrin positive. The contents of tubes 8-17 were combined and concentrated in vacuo to about 1 mL. The concentrate was treated with 3 mL of MeOH and filtered. The filter was washed with 0.5 mL of 75% aqueous MeOH. The filtrate and washings were combined, diluted with an additional 10 mL of MeOH and left at room temperature overnight. The amorphous precipitate was filtered, and crystalline embryos were seeded into the filtrate to give Compound IV as colorless crystals, which was identical to the authentic sample. The filtrate was evaporated in vacuo and lyophilized. The yield and experiment of each fraction are shown in the following table.

IV. Quantity Test Yield B. subtilis K. pneumoniae B. subtilis K. pneumoniae

Amorphous 70 mg 737 / mg 927 / mg 0.9% 1.1% powder

Crystals 1098 mg 1163 "1087" 19 JC 19%

Lyophilized 180 mg 608 "50U" 1.9 ^ 1.6% powder BB-K 11 is 3 "-N-ZL - (- J-TT-amino-c <-hydroxybutyryl / -canamycin A.

BB-K 29 is 3-N- [L - (-) - N-amino- (N-hydroxybutyryl) -kanamycin A.

The starting material was prepared as follows: Preparation of N-hydroxy-5-norbornene-2,3-dicarboximide ester (XX) of L-N-benzyloxycarbonylamino-d.-Hydroxybutyric acid Dicyclohexylcarbodiimide (DDC) (2.06 g, 0.01 mol) was added to a stirred solution of Ly-benzyloxycarbonylamino-β-hydroxybutyric acid (2.53 g, 0.01 mol) and compound XIX (1.79 g, 0.01 mol) in 50 ml of dry THF (tetrahydrofuran) at 10 ° C. . The mixture was stirred overnight at room temperature. The precipitated dicyclohexylurea was removed by filtration and washed with THF.

The filtrate and washings were combined and evaporated in vacuo. The oily residue (k, 5 g) was passed through a column of silica gel and eluted with ethyl acetate to give H.0 g (96%) of active ester XX as a colorless oil. Infrared spectrum (IR, Neat): 3300, 1820, 17O, 1735, 1720, 1695 cm-1. Nuclear Magnetic Resonance Spectrum (NMR) (CDCl 3 + 1 drop of D 2 O): δ (ppm) 1.63 (2N, q, J = 9Hz), 2.15 (2H, m) 3, HO (6H, m), H, 60 (1H, dd, J = 7 and 5 Hz), 5.13 (2H, s), 6.20 ( 2H, broad), 7, H0 (5H, s).

9,60217

Example 2 Preparation of 1-N- [L - (-) -] -? - amino- [X-hydroxybutyryl] canamycin A (IV)

To a stirred solution of 6'-N-Cbz-kanamycin (II) (1.36 g, 2.2 mmol) in 60 mL of 50% aqueous THF was added dropwise at room temperature over 30 minutes 930 mg (2 mmol) of active ester XX in 10 mL of THF. The mixture was stirred overnight and then hydrogenated under normal pressure at room temperature with 10% palladium on carbon (1.0 g). The catalyst was removed and washed with water. The filtrate and washings were combined, the organic solvent was evaporated in vacuo. The resulting aqueous solution was applied to a CG-50 column (NH 4, 70 mL) eluted sequentially with 0.1+ liters of water, 0.5 L of 0.1 N NH 4 OH, 0.5 L of 0.3 N NH 4 OH, with 1 liter of 0.5 N NH 4 OH and finally with 0.5 liter of 1 N NH 4 OH. The eluate was collected in 20 ml fractions and divided according to the Rf value (TLC on a silica gel plate, S-110, ninhydrin) and B sublitis based on disc experiments using PCI 219 and K. pneumoniae type 223038 A 20680. Each fraction was evaporated in vacuo and lyophilized.

Example 3 In situ preparation of 1-N-ZL - (-) -?

In a suitable apparatus, 1000 g (1.07 * + moles) of 6'-carbobenzoxy-1,3,3 "-trisalicylalanicamycin A in 11 L + 00 ml of THF were slurried at 22-25 ° C. 181.5 g (0.716 moles) of 1 + -benzyloxycarbonylamino-2-hydroxybutyric acid and 0.12 moles of N-hydroxy-5-norbornene endo-2,3-dicarboximide (ΧΙχ) were added to the solution. The pH ranged from 5.5 to 6.0 The temperature was maintained at 22-25 ° C, and 162.3 g (0.787 moles) of dicyclohexylcarbodiimide dissolved in 3000 ml of dry THF was added to the mixture. At -25 ° C, then cooled at 0-5 ° C for 2 hours to crystallize the dicyclohexylurea.

The urea was removed by filtration and the precipitate was washed with 800 ml of 95/5 THF-water. The filtrate and washings were combined and evaporated under reduced pressure to 3000 ml.

1 + 000 ml of methanol were added and the mixture was again evaporated in vacuo to 3000 ml. To the solution thus obtained was added 2000 ml of absolute methanol at 1 + 0 ° C, then the solution was inoculated with 6'-carbobenzoxy-1,3,3 "-trisalicylalicamycin A and allowed to cool and crystallize for 0.5-1 hours. 2000 ml of water over 1/2 hour and stirred well for 2 hours at about 22-25 ° C.

The temperature was then lowered to 0-5 ° C for one hour.

10 6021 7

The precipitate was removed by filtration and washed with 900 ml of cold 2: 1 methanol-water mixture and then with 800 ml of methanol. The filtrate and washings were collected. The precipitate was mainly recovered 6'-carbobenzoxy-1,3,3 "-trisalicylalanine mycin A.

The filtrate and washings obtained above were diluted with 1000 ml of water. To this was added 2000 ml of methylene chloride. The pH was adjusted to 1.8-2.0 at 25 ° C with 6N HCl. This pH was maintained for 1/2 hour.

The methylene chloride layer was separated and the aqueous layer was washed with an additional 2000 mL of methylene chloride. The combined organic phases are recovered to recover any salicylaldehyde.

The pH of the aqueous phase is raised to 3.5 ~> 0 with concentrated ammonium hydroxide.

2 This solution was hydrogenated at room temperature under a pressure of 3.5 kg / cm 3 with hydrogen gas in the presence of 5 # Pd / C. The bioavailability of the desired product in solution was 30-5% of theory.

Preparation of N-hydroxy-5-norbornene endo-2,3-dicarboximide (HONB) (XIX) as starting material

Preparation was performed according to L. Bauer and S.V. By the method of Miarka (J. Org. Chem., 2U, 1293, (1959)). 5-Norbornene endo-2,3-dicarboxylic anhydride (115 g, 0.0 M moles) was added to a hydroxylamine solution prepared by adding * + 7.5 g (0.1 + 5 moles) of sodium carbonate to hydroxylamine hydrochloride (60.5 g , 0.87 moles) in a solution in 1 x 0 ml of water. The mixture was heated for one hour at 60-70 ° C and allowed to stand in a refrigerator overnight. The crystals were filtered off, washed with 120 ml of cold 5N HCl and dried to give 8.9 g (70%) of compound XIX, m.p. 172 ° C. The individual solution was acidified to pH 2 and extracted with CHCl 3 (70 mL x 10). Evaporation of the CHCl 3 extracts gave a second crop of XIX (7.2 g, 6 #), m.p. 170-172 ° C.

Example U

Preparation of compound IV using various amounts of N-hydroxy-5-norbornene-2,3-dicarboximide while keeping the other molar #s constant Using the N-hydroxy-5-norborneene-2,3-dicarboxyimide used therein in the process of Example 3. instead of the amount of dicarboxyimide (0.12 moles of HONB), the following varying mole percentages of the compounds were as follows:

Experiment_Mole- #HONB_Yield of Compound IV # 1 0 9.33 2 2 20.07 3 10 31.72 k 50 36.52 5 100 36.01 11 6021 7

It can be seen from the above results that compound IV is obtained in the reaction completely without HONB. However, it can also be seen that the best yields of compound IV are obtained when the molar amount of HONB is 10-50% of the molar amount of 1 * -benzyloxycarbonylamino-2-hydroxybutyric acid used in the acylation reaction. The exact mechanism of the acylation reaction of protected kanamycin A is unknown. However, it has been found that the presence of H0NB for some reason produces better yields than previously obtained.

Claims (2)

A process for the preparation of 1- / L - (-) - Y-amino-c * -hydroxybutyryl7-kanamycin A of the formula HO CVNH2 H °> 4 H ° 1 NH_ ° VK ch2oh ho -i-7 \\ H0-I / - NH 'y' * - o / c = o HN --1- \ / ΤΆ / HO \ / CH2 ° CH? m2 characterized in that the compound of the formula H ° ho ch2-nco-ch2-c6h5 H ° ^ —7 \\ 111 H ° A z CHpOH HO ^ —— ho> L * -Wa / “V 13 £ 0217, wherein Z is a group - »= 0 ^ 0, -„ ^ 0-och3, HO H, / Λ · H -N = C- ^ y or -N = CC -CH3 \ = / Sv'CH3 is reacted in situ in at least a molar ratio of 1 : 0.5 with a compound of formula 0 2 2 I) N-O-C-CH-CH 2 -CH 2 -NH-C-O-CH 2 -CgH XX OH 0 and the reaction product is hydrogenated in situ to give a compound of formula IV. 6 0 21 7 1U For use in the preparation of 1- / L - (-) - * N '- amino-o-hydroxybutyryl7-kanamycin A with the formula HO-K PH2-NH2 HO \ NH V_ ch2oh ho -j-7 \\ HO— ^ -o / c = o 7 "Λ / HOiH„ 0 V c „m2 kännetecknat därav, att main omsätter föreningen med formeln HS CHo-NC-0-CHo-C £ Hc HO -V 2 2 6 5 HO 0- CH2OH ho -j - tO \ / ^ zz / HO 0