DK146778B - ANALOGY PROCEDURE FOR PREPARING SEMI-SYNTHETIC OLEANDOMYCIN COMPOUNDS OR NOT TOXIC ACID ADDITION SALTS THEREOF - Google Patents

Description

(19) DENMARK \ rbJ

f (i2) PUBLICATION MANUAL (n) 146778 B

Dl The Patent and Trademark Office

(21) Patent Application No. 0571/77 (51) IntCI.3: C 07 H 17/08 (22) Filing Date: 10 Feb 1977 (41) Alm. available: 04 Sep 1977 (44) Submitted: 02 Jan 1984 (86) International Application No: - (30) Priority: 03 Mar 1976 US 663467 08 Jul 1976 US 703464 29 Jul 1976 US 709703 (71) Applicant: 'PFIZER INC .; New York, US.

(72) Inventor: Frank C. SciavolIno; US.

(74) Plenipotentiary: Hofman-Bang & Boutard Patent Office (54) Analogous Process for the Preparation of Semi-Synthetic Oleandomycin Compounds or Non-Toxic Acid addition Salts thereof

BACKGROUND OF THE INVENTION 1. Field of the Invention This invention relates to an analogous method for the preparation of novel semi-synthetic oleandomycin compounds having the general formula II or acid addition salts thereof set forth in claim 1, which compounds are useful as antibiotics for the treatment of bacterial infections, such as oleandomycin. fermentation liquids and its use as an antibacterial agent were first described in United States Patent Specification No. 5,197,117.

2 757 123 to which reference is made. The naturally-occurring compound is known to have the structural formula 2

OrV f: l H0> 10

"N'-fY

ii

o 0H

OCH,

D

The conventionally accepted numbering is shown at a number of different positions. Several synthetic modifications of this compound are known, especially those in which from 1 to 3 of the free hydroxy groups found at the 2'-, 4'- and 11-positions are esterified as acetyl esters. U.S. Patent No. 3,022,219 discloses other modifications in which the acetyl group of the above esters is replaced by another, preferably unbranched, alkanoyl group having 2-6 carbon atoms or trifluoroacetyl. Semi-synthetic oleandomycins are also known in which one or more of the hydrogen atoms in the above hydroxy groups is replaced by a tri (lower alkyl) silyl group and preferably trimethylsilyl.

The semi-synthetic oleandomycin compounds of this invention, having a methylene group at the 8-position, are generally more active against Gram-positive microorganisms than against Gram-negative. They provide protection against Staph in vivo. aureus infections comparable to natural oleandomycins and their antibacterial activity in vitro against a wide range of organisms are comparable to the activity of the commercially available semi-synthetic compound triacetyl-oleandomycin while exhibiting a surprisingly high activity against the patho -genic Gram-negative organism Neisseriae sicca 66C000.

The process according to the invention is characterized by the characterizing part of claim 1.

Acetone, an alkanol of 1-4 carbon atoms or tetrahydrofuran is preferred as reaction-inert solvent and CrCl 2 is the preferred reducing agent.

The process of the invention can be carried out by adding a solution of chromium (II) chloride, prepared by treating a hydrochloric acid solution of chromium (III) chloride with zinc dust and mercury (II) chloride, to a flask under a carbon dioxide atmosphere under simultaneous addition of an acetone solution of a compound of formula I IV (see claim 1), wherein R, R and R have the meanings given in claim 1, and are preferably hydrogen and / or acetyl. Additions are made within 10 - 20 minutes. After stirring at room temperature for 20 - 40 minutes, water and ethyl acetate are added and stirring is continued for another 10 - 20 minutes. The ethyl acetate phase is separated, washed with water and sodium hydrogen carbonate solution at pH 8.5, dried and concentrated in vacuo to give a compound of formula II.

A compound of formula II having a trialkylsilyloxy group of 1-6 carbon atoms in the alkyl moieties at 11 position may also be prepared by bringing a compound of formula II wherein R 2 is hydrogen and R 2 and R 2 are alkanoyl of 2 to 3 carbon atoms. , in contact with a corresponding trialkylsilyl chloride in the presence of imidazole and dimethylformamide at room temperature. After adding water and ethyl acetate and adjusting the pH to 10.5, the organic phase is separated, washed, dried and evaporated under reduced pressure.

A compound of formula II wherein R, R 2 and R 2 are alkanoyl of 2-3 carbon atoms can be converted to the corresponding compound, which is hydrogen, by selective hydrolysis of the alkanoyloxy group in the 4 "position, f. for example with sodium isopropoxide in isopropyl alcohol.

The pharmaceutically acceptable acid addition salts of the obtained semi-synthetic oleandomycins can be prepared by contacting a solution of a compound of formula II in a solvent such as acetone with a stoichiometric equivalent of a mineral acid such as hydrogen chloride, hydrogen bromide, phosphoric acid or sulfuric acid. , an organic acid selected from aspartic acid, citric acid, tartaric acid, gluconic acid, succinic acid and stearic acid, or an alkyl sulfuric acid such as lauryl sulfuric acid. The salt is precipitated after neutralization or, if necessary, after partial evaporation of the reaction solution. The product can be recovered by filtration, centrifugation or lyophilization.

The oleandomycin compounds prepared by the process of the invention are effective in inhibiting the growth of microorganisms, especially Gram positive microorganisms. The high activity against Gram-positive organisms exhibited by these compounds is in some cases in contrast to the low activity against Gram-negative organisms. The following table illustrates the in vitro antibiotic spectrum of the compounds of Formula II compared to the known compound triacetyloleandomycin (TAO). The tests were performed according to the "minimum inhibitory concentration" (MIC) procedure as described by Ericsson and Sherris / E.Fi. Ericsson and J.D. Sherris, Acta. Pathol. Microbiol. Scand. Suppl., 217B 64 (19711? · 5 145778 MIC values (yug / ml) for some semi-synthetic oleandomycins.

N (CH3) 2 R20 / "r r

..... / Y

J 0¾ 6ch3 ¥ (ch3) 2 ”% γΥ.

n TAO

° (L 1 /// oi> uvv'v j OAc and 3666778

Compounds of Formula II R = Ac

Microorganisms TAO _ _ Ac Ac Ac _ * £ »3« l e r (1) Staph. aur. 01A005 3.12> 200 3.12 25 200 (2) 01A052 3.12> 200 3.12 50 200 (3) 01A109R> 50> 200> 200> 200> 200 (4) 01A110R> 50> 200> 200> 200> 200 (5) 01A111R 3.12 25 3.12 3.12 0.78 (6) 01A087RR> 50> 200> 200 -> 200 (7) 01A400R 6.25> 200> 200 200> 200 (8) Strp.fae. 02A006 6.25> 200 12.5 200> 200 (9) Strp.pyog. 02C203 0.78 6.25 0.78 25> 200 (10) 02C040R> 50 - - (11) Myco.smeg. 05A001 25 “-200 -> 200 (12) B.sub. 06A001 1.56> 200 3.12 - 6.25 (13) E.coli 51A229> 50 *> 200> 200> 200> 200 (14) 51A266> 50 *> 200> 200> 200> 200 (15) 51A125R> 50 *> 200> 200> 200> 200 (16) 52A104> 50 *> 200> 200> 200> 200 (17) Klebs.pn. 53A009> 50 *> 200> 200> 200> 200 (18) 53AQ31R> 50 *> 200> 200> 200> 200 (19) Prot.mira. 57C064 -> 200> 200> 200> 200 (20) Prot.morg. 57G001> 50 *> 200> 200> 200> 200 (21) Salm.chol.su. 58B242> 50 *> 200> 200> 200> 200 (22) S L typhm. 58D009> 50 *> 200> 200> 200> 200 (23) 58D013C> 50 *> 200> 200> 200> 200 (24) Past.multo. 59A001 50> 200 50> 200> 200 (25) Serr.amr. 63A017> 50 *> 200> 200> 200> 200 (26) Ent.aero. 67A040> 50 *> 200> 200> 200> 200 (27) Ent.cloa. 67B003> 50 *> 200> 200> 200> 200 (28) Neiss.sic. 66C000 50 3.12 0.20 3.12 1.56 rk

The tests were only conducted at 50 µg / ml, but previous results show no activity at the 200 µg / ml level.

In the table, the effect of Compounds II is compared to the action of the commercially available oleandomycin derivative, triacetyloleandomycin (TAO), against a variety of Gram-positive (Nos. 1-12) and Gram-negative (Nos. 11-28). microorganisms. It is seen that compounds II have an activity of the same size as TAO to most organisms and a much higher activity to the pathogenic Gram-negative organism Neisseriae sicca 66C000.

For effective prophylactic and anti-infectious use in vivo, the semi-synthetic oleandomycin compounds produced by the method of the invention may be administered either alone or in combination with a pharmaceutically acceptable carrier and by both oral and parenteral routes. The choice of route of administration and dose is ultimately made by the treating physician and the choice is based on the individual patient's condition. However, the usual dosage for administration to humans may be in the range of 500-2000 mg per day. per day, and preferably in 1-4 doses. However, this dosage may vary somewhat with the individual's weight; In general, from about 10 to about 40 mg / kg of body weight per day can be used. day.

For pharmaceutical use, the compounds can be combined with inert pharmaceutical excipients such as lactose, mannitol and starch, and formulated in dosage forms such as tablets, capsules and the like. For parenteral administration, the compounds can be formulated with an inert, parenterally acceptable medium such as water, saline, sesame oil, propylene glycol and the like. These various pharmaceutical dosage forms are prepared by well known pharmaceutical methods.

The process according to the invention is more fully illustrated by the following embodiments.

EXAMPLE 1 1 8,8a-Deoxy-8,8a-Methylene-2 1-4ll-diacetyloleandomycin

A 5 liter 3 neck round bottom flask was charged with zinc dust (200 g) and mercury (II) chloride (20 g). After the solids were well mixed, 1N hydrochloric acid (500 ml) was added and the mixture was stirred vigorously for 15 min. The aqueous supernatant was removed, fresh IN hydrochloric acid (500 ml) was added and the flask was placed under a carbon dioxide atmosphere. A filtered solution of chromium (m) chloride (1 kg in 1300 ml of 1N HCl) was quickly added to the zinc amalgam. The mixture was stirred under a carbon dixoid atmosphere for 1 hour, during which a light blue color was developed which showed the presence of chromium (II) chloride (CrCl 2). Stirring was stopped after 1 hour and the zinc amalgam was allowed to settle on the bottom of the flask. ..

A solution of 2 ', 4 "diacetyloleandomycin hydrochloride (500 g) in acetone (3.5 l) and water (1.75) was placed in a dropping funnel on a 12 l 3-necked round bottom flask equipped with a mechanical stirrer. To this flask was added, under a carbon dioxide atmosphere and with stirring, the solution of 2 ', 4 "-diacetyloleandomycin hydrochloride and the pre-prepared solution of chromium (II) chloride. The solutions were added simultaneously at such a rate that both were added at the same time. The addition took about 12 min.

After stirring at room temperature for 35 min, water (2 L) and ethyl acetate (2 L) were added to the reaction mixture and stirring was continued for 15 min. The ethyl acetate layer was separated and washed with water (1600 ml). The ethyl acetate was separated and the aqueous extracts combined and washed with fresh ethyl acetate (2 L). The ethyl acetate layer was again separated and washed with water (1.7 L). The organic phase was separated and the aqueous washing solutions combined and treated with sodium chloride (1500 g). The additional ethyl acetate which was separated was suctioned off and combined with the other ethyl acetate extracts. Water was added to the combined ethyl acetate extracts and the mixture was adjusted to pH 8.5 with sodium bicarbonate. The organic layer was separated, washed with water, saturated sodium chloride and dried over anhydrous sodium sulfate. Filtration and evaporation of the solvents under reduced pressure gave a white solid which was recrystallized from ethyl acetate / heptane to give the above compound (237 g, mp 184-186 ° C).

element Analysis

Calculated: C 61.96, H 8.67, N 1.85 Found: C 61.78, H 8.54, N 1.87.

spectral

NMR (CDCl1): Y = 5.61 (1H) bS 5.53 (1H) bS 3.36 (3H) δ

J '2.26 (6H) S 2.10 (3H) S 2.06 (3H) S

IR (CHCl 3): 5.75 5.90 and 6.15 µm UV (CH 3 OH): 225 nm < 5338 l / mole EXAMPLE 2 8,8a-deoxy-8,8a-methylene-2 ', 4 ", 11-triacetyloleandomycin

A solution of 2 ', 4 ", 11-triacetyloleandomycin (500 g) in acetone (3.25 L) and water (1.75 L) was treated with a solution of chromium (II) chloride / prepared as described in Example 1 from chlorine (III) chloride (1000 g), zinc dust (200 g) and mercury (II) chloride (20 g // in a carbon dioxide atmosphere as described in Example 1. The final ethyl acetate extracts (2500 ml) were concentrated to 800 ml, 3 volumes of heptane were added and the crystallization allowed to proceed overnight to give the above compound in 2 aliquots (426 g, 132-134 ° C).

element Analysis

Calculated: C 61.71, H 8.46, N 1.76 Found: C 61.59, H 8.42, N 1.72.

146778 ίο

spectral

NMR (CDCl3): δ = 5.86 (1H) bS 5f61 (1H) bS 3.33 (3H) δ

J 2.26 (6H) S 2.06 (6H) S 2.01 (3H) S

IR (CHCl 3): 5.70 5.90 6.00 and 6.17 µm UV (CH 3 OH): 228 nm £ = 5927 l / mole EXAMPLE 3 8,8a-deoxy-8,8a-methylene-21-acetyloleandomycin

A solution of 2'-acetyloleandomycin (29.2 g) in acetone (200 ml) and water (100 ml) was treated with a solution of chromium (II) chloride / prepared from chromium (II) chloride (50 g), zinc dust (10 g) and mercury (II) chloride (1 g) by the method of Example 1 / in a carbon dioxide atmosphere as described in Example 1. The final ethyl acetate extracts were concentrated to dryness under reduced pressure and the residue crystallized from ether. petroleum ether followed by recrystallization from ethyl acetate / petroleum ether to give the above compound (8.4 g, m.p.

183.5-185 ° C).

element Analysis

Calculated: C 62.16, H 9.02, N 1.95 Found: G 61.97, H 8.91, N 2.01.

spectral

NMR (CDCl3): (f = 5.63 (1H) bS 5.58 (1H) bS 3.43 (3H) S

2.36 (6H) S 2.08 (3H) S

IR (CHCl 3): 5.78 5.90 5.95 and 6.12 µm UV (CH 3 OH): 224 nm £ = 4468 µmol / mole EXAMPLE 4 8,8a-deoxy-8,8a-methylene 2l, 11-diacetyloleandomycin

To a stirred solution of 8,8a-deoxy-8,8a-methylene-2 ', 4 ", 11-triethylethyleandomycin in isopropyl alcohol (50 ml) at room temperature under nitrogen was added a solution of sodium isopropoxide in isopropyl alcohol (17.8 ml After 3.5 hours, another portion of sodium isopropoxide solution (17.8 ml, 0.176 M) was added and stirring was continued for 1 hour at room temperature. The pale yellow solution was poured into water and extracted with ethyl acetate. was separated, washed with water and saturated sodium chloride solution, dried over sodium sulfate, filtered and evaporated The resulting foam was crystallized from ethyl acetate / hexane to give the above compound (2.35 g, mp 117-119 ° C).

spectral

NMR (CDCl3): δ = 5.96 (1H) bS 5.73 (1H) bS 3.45 (3H) δ

J 2.33 (6H) S 2.10 (3H) S

EXAMPLE 5 8,8a-Deoxy-8,8a-Methylene-21,4 "-diacetyl-11-trimethylsilyloleandomycin In a flame-dried 100 ml 3-neck round bottom flask fitted with magnetic stirrer and dryer tube, a suspension of 8,8a-deoxy-8 , 8α-methylene-2 ', 4 "-diacetyloleandomycin (20 g) and imidazole (4.5 g) in dry dimethyl fomamide (32 ml) treated with trimethylsilyl chloride (6.72 ml) and stirred at room temperature for 90 min. The pale yellow solution was poured into water (300 ml) and ethyl acetate (300 ml) and adjusted to pH 10.5 with 4N sodium hydroxide solution. The aqueous phase was separated and the ethyl acetate layer washed with water (300 ml) and saturated sodium chloride solution (300 ml), dried over anhydrous sodium sulfate and evaporated under reduced pressure to a white foam (22 g), which was homogeneous by thin layer chromatography.

Claims (2)

14677δ 12 (silica gel plates eluted with ethyl acetate / acetone 3: 1, 0.6; carbon tetrachloride / diethylamine 9: 1, R f 0.7). Spectral data NMR (CDCl3); <$ = 6.00 (1H) bS 5.73 (1H) bS 3.36 (3H) S J 2.33 (6H) S 2.13 (6H) S 0.16 (9H) S P a t e n t k r a v: An analogous process for the preparation of semi-synthetic oleandomycin compounds of formula I II Y "- OCH3 wherein R is alkanoyl of 2-3 carbon atoms, R, is hydrogen or alkanoyl of 2-3 carbon atoms and R2 is hydrogen, alkanoyl with 2-3 carbon atoms or trialkylsilyl having 1-6 carbon atoms in the alkyl moiety, or non-toxic acid addition salts thereof, characterized in that a compound of the formula