JPH0149356B2 - - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention provides 9-deoxo-
9a-aza-9a-related to novel derivatives of homoerythromycin A and its 4″-epimer, processes for their production and compositions. For details, see 9-deoxo-9a
-Aza-9a-Relates to cyclic ether derivatives of homoerythromycin A and its epimers, and further relates to pharmaceutically suitable acid addition salts thereof, compositions of the above-mentioned compounds as antibacterial agents, and processes for their preparation. Erythromycin A is a macrolide antibiotic produced by fermentation and is described in US Pat. No. 2,653,899. A number of derivatives of erythromycin A have been produced to modify its biological and/or pharmacokinetic properties. mono-
and esters of erythromycin A with dicarboxylic acids, Antibiotics Annual, 1953-
1954, Proc.Symposium Antibiotics
(Washington, DC) pages 500-513 and 514
−Reported on page 521. US Patent No.
No. 3,417,077 describes cyclic carbonate esters of erythromycin A, reaction products of erythromycin A, and ethylene carbonate as active antimicrobial agents. U.S. Pat. No. 4,328,334, issued May 4, 1982, describes 9-deoxo-9a-aza-9a-homoerythromycin A and 11-aza-10-deoxo-10.
- Described as dihydroerythromycin A. The above-mentioned compounds are ring-expanded (homo) derivatives of erythromycin, and additionally contain nitrogen (aza) in the ring system.
Since it is a compound into which an atom has been introduced, the parent ring of the compound of the present invention includes 9-deoxo-9a-aza-9a-
Homoerythromycin A is well named. Belgian patent number issued June 1, 1982
892357 and its UK application 2094293A (1982
9-deoxo-9a-
describes the N-methyl derivative of aza-9a-homoerythromycin A, which was published in 1982.
U.S. Patent Application No. 399,401, filed July 19, 2006, seeking priority from U.S. Patent Application No.
No. 441981 (filed on November 15, 1982) is in dispute. U.S. Pat. No. 4,382,085, issued May 3, 1983, describes 4-eperythromycin A or 4″-
Describes compounds in which the OH group has an axial bond. The 4″-OH group of erythromycin A is an equatorial bond. Certain cyclic ethers of 9-deoxo-9a-aza-9a-homoerythromycin A and its 4″-epimer are potent against Gram-positive and Gram-negative bacteria. It turned out to be an antibacterial agent. The compound has the formula (). where n is 1, 2 or 3; the wavy line in the 4"-position is generic and includes both epimers. When n is 1, the 4"-hydroxy group is an equatorial bond. When n is 2 or 3, the epimeric forms of the above-mentioned compounds differ structurally only in the bonding of the chiral center in the 4''-position, ie the 4''-OH group is an axial and equatorial bond. An axial bond is represented by a solid line or a wedge-shaped bond with an OH group at the 4″-position, and an equatorial bond is represented by a dashed line. “Axial” and “equatorial” as used here refer to the 4″-hydroxyl and Represents two possible chiral epimers for hydrogen. The present invention also encompasses pharmaceutically suitable acid addition salts of the above-mentioned compounds, which are useful for the same purpose as the compounds of formula. These salts include, but are not limited to, the salts listed below, such as hydrochloride, hydrobromide, sulfate, phosphate, formate,
acetate, propionate, butyrate, citrate,
Glycolate, lactate, tartrate, malate, maleate, fumarate, gluconate, stearate, mandelate, pamoate, benzoate, succinate, p-toluenesulfonate , aspartate. The scope of the present invention also includes a method for producing the formula. Intermediates useful in its preparation are represented by the following formulas and: In the formula, R ₁ is -(CH ₂ ) _p CN or -(CH ₂ ) _n -
NH ₂ ; p is 1 or 2; m is 2 or 3; The formula and the wavy line at each 4"-position in the formula represent both epimers, i.e. axial and equatorial bonds at that position. The present invention further relates to the by-product (n Also falling within that scope are compounds of formula (), formed as 3). The wavy line at the 4″-OH group in the formula represents an axial and equatorial bond at that position. , Staphylococcus aureus ( Staphylococcus
aureus), Streptococcus pyogenes and Crum-negative bacteria,
For example, Pasturella multocida ( Pasturella
multocida), Neisseria sitsuka ( Neisseria
sicca ) in vitro ,
It has effective antibacterial properties. Furthermore, the compound of formula () is derived from Neisseria gonorrhea ( Neisseria
gonorrhea) and Haemophilus
It shows excellent activity against many Gram-positive and Gram-negative bacteria in vitro and in vivo against many Gram-positive and Gram - negative bacteria. In terms of oral anti-infective activity, compounds of formula () are similar to 9-deoxo-9a-methyl-9a-aza-9a-homo-erythromycin A and are effective in vivo.
The corresponding 9- with no actual oral activity in vivo)
It is completely different from deoxo-9a-aza-9a-homoerythromycin A. Compounds of formula () where n is 3 can be prepared by cyanoethylation of the appropriate 9-deoxo-9a-aza-9a-homoerythromycin A epimer.
Acrylonitrile is used in excess and is used as both reactant and solvent. The reaction is generally carried out at the reflux temperature of the acrylonitrile, about 77°C, until the reaction is complete. For example, the reaction can be carried out using a low temperature such as 50°C, but it is better not to react at a low temperature because the reaction time will be longer. The reaction is carried out at a higher temperature using a reaction inert solvent other than excess acrylonitrile, that is, a solvent that does not enter the reaction system together with the reactants and/or products and does not adversely affect the yield of the desired cyanoethyl derivative. I can do it. Of course, the reaction-inert solvent can be used at any temperature at which the reaction will proceed. Suitable solvents are C _1-4 alkanols; cyclic or acyclic ethers such as dioxane, tetrahydrofuran, diethyl ether, diethylene glycol dimethyl ether; aromatic hydrocarbons such as benzene, toluene, xylene. However, it is a good practice to use excess acrylonitrile as solvent. The reaction progresses using a silica gel plate.
Perform thin layer chromatography (TLC) developed with CH ₂ Cl ₂ /CH ₃ OH/concentrated NH ₄ OH (6/1/0.1), spray with vanillin/ethanol/H ₃ PO ₄ and heat, and check. do. The solvent is evaporated and the product is obtained as a colorless foam. For convenience, this can be used as is in the next step. At T.L.C.
This indicates that it is essentially a monocyanoethylated product. The presence of trace amounts of polar products was also detected by TLC. The thus obtained 9a-cyanoethyl derivative () is converted into the corresponding 9a-(gamma-aminopropyl) derivative () by hydrogenation with Raney nickel according to the method described below. This is reacted with at least a stoichiometric amount of isoamyl nitrite and glacial acetic acid according to the method described below to convert it into the corresponding 7-membered ring, cyclic ether. At this time, the corresponding 9a-(gamma-acetoxypropyl) derivative, a compound of formula (), is obtained as a by-product. Compounds of the formula where n is 2 are suitable, 9-
Deoxo-9a-aza-9a-hydroxy-9a-homoerythromycin A3′-N-oxide 4″-
The OH epimer is reacted with excess bromoacetonitrile in a reaction inert solvent from 20°C to 100°C (room temperature is preferred).
Produced by alkylation. Suitable solvents are halogenated hydrocarbons such as methylene chloride, chloroform; benzene, toluene, xylene,
Aromatic hydrocarbons such as; ethers such as diethylene glycol dimethyl ether, dioxane, tetrahydrofuran, and diethyl ether. The reaction is carried out in the presence of an acid acceptor, for example a C _1-4 trialkylamine, an organic base such as pyridine, an inorganic base such as an alkali _metal carbonate or bicarbonate. The acid acceptor is used in equivalent amounts to the excess bromoacetonitrile used to achieve optimal conversion of the macrolide reactant to product. Hydrogenation () in the presence of Pd/C in an inert solvent such as ethanol to remove the 9a- and 3'-oxides yields a cyanomethyl compound of formula () where p is 1. The thus obtained 9a-cyanomethyl derivative (formula ()) is reduced with sodium borohydride in the presence of cobalt chloride in a solvent with or without a hydroxy group at room temperature to form the corresponding 9a-(beta-aminoethyl). Obtain the derivative. In this reaction,
A preferred molar ratio of CoCl ₂ to NaBH ₄ is 1:5. In practice, _NaBH4 should be added in excess, e.g.
10 moles of NaBH ₄ are used for every mole of substrate. 9a
Reaction of the -(beta-aminoethyl) derivative with at least a stoichiometric amount of amyl nitrite/glacial acetic acid gives the corresponding 6-membered cyclic ether. 5-membered ring of formula (), cyclic ether (n=1,
4'-OH is equatorial coordination) is reacted with 9-deoxo-9a-aza-9a-homoerythromycin A with 1-4 equivalents of formaldehyde (conveniently a 37% aqueous solution) and formic acid in an inert solvent at room temperature. It is produced by reacting with Typical solvents are halogenated hydrocarbons such as methylene chloride, chloroform and carbon tetrachloride. This reaction is limited to relatively low reaction temperatures. The reaction proceeds easily at about 15°C to 30°C (20°C to 25°C is better). For example, a 9a-methyl derivative is obtained when the reaction is carried out at a high temperature such as reflux temperature.
That is, reductive methylation occurs. Methylation occurs exclusively at high temperatures of about 50°C. However, under the reaction conditions of the present invention, cyclic ether formation occurs between the 11beta-OH and the 9a-aza group. Compounds of formulas (), () and (); R ₁ is -
Compounds with 2′ and/or 4″-hydroxy groups in (CH ₂ ) _p CN are described, for example, by Jones et al., J.Med.Chem. 15 ;
631 (1972); Banaszek et al. Rocy.Chem. 43 , 763
(1969), the corresponding C _2-3 alkanoyl derivatives can be prepared by conventional acylation. The 2′- and 4″-hydroxy groups are acylated by reaction with a suitable acid anhydride (e.g. (R ₂ CO) ₂ O) in pyridine. Solvolysis of the 2′,4″-ester with methanol A mixed ester, e.g. 2'-acetyl-4"-propionyl, is prepared by reacting the 4"-ester (R ₃ = propionyl) in an inert solvent with the mixed ester of Jones et al. (supra). The acid addition salts of the present invention are prepared by reacting a compound of the formula with at least an equivalent amount of the appropriate acid in an inert reaction solvent. In the case of hydrochloride, it is reacted with pyridine hydrochloride.Since one or more basic groups are present in the compound of the formula, a sufficient amount of acid is added to form a polyacid which is acid-added with each basic group. Addition salts can be produced.When producing acid addition salts of compounds of the formula in which 2'-OH is acylated, isopropanol is used as a solvent to prevent solvolysis of the alkanoyl group.Acid addition salts are If it is insoluble in the reaction inert solvent, it can be obtained by precipitation by adding a non-solvent to the acid addition salt or by distilling off the solvent.Various Gram-positive bacteria and certain Gram-negative bacteria,
For example, for spherical or oval (coccus) bacteria, the formula (),
Sensitive to compounds () and ().
In vitro activity is readily determined in in vitro assays against various bacteria in brain-heart infusion medium using the 2-fold dilution method. Their in vitro activity makes them useful for topical application in the form of ointments, creams; for sterilization of hospital equipment, etc.; for use as antimicrobial agents in industrial applications such as water treatment, sludge treatment, paint and wood preservatives. Become. For in vitro use, such as topical application, by methods known to those skilled in the art.
The selected compounds may conveniently be formulated into lotions, ointments, creams, or gels. For this purpose, it is common to use active ingredients at concentrations of about 0.01 percent to 10 percent (by weight) of the total ingredients. The method of administration is to apply any amount to the site of infection, generally at least once a day. The compounds of the present invention (compound-) and conventional compounds (compound-) against various microorganisms are shown below.
Comparison of minimum inhibitory concentration (MIC) (mg/ml) with [Table] In addition, the compound and the formula () (in the formula n
=3) is the equatorial epimer and axial epimer of the compound. Table 1 shows that the compounds of the invention have particularly pronounced antibacterial properties against Staphylococcus aureus, E. coli and H. influenzae. Table 2 below shows the results of pharmacodynamic testing of the compound, , in experimental animals. [Table] [Table] * Maximum blood concentration
** Area under the curve obtained by plotting blood concentration over time
The above table shows the large increase in half-life as an indication of the unexpectedly high utility of the compounds of the invention. This is particularly important with respect to the major microorganism Haemophilus influenzae. The above table also shows that the excellent pharmacodynamic properties of the compounds are also unexpected. It is not possible to predict from the antibacterial spectrum of the compound the fact that the compound of formula () exhibits an antibiotic activity equivalent to that of the compound. Furthermore, the Cmax and AUC values of the compounds of the present invention in Table 2 are generally smaller than those of conventional compounds, and blood concentrations do not become higher than necessary. That is, it should be noted that the compounds of the present invention can prevent acute toxicity due to increased blood concentration. As described above, the compounds of the present invention have highly desirable pharmacodynamic properties such that moderate blood concentrations can be maintained for a long period of time. Additionally, compounds of the formula of the present invention may be used in Gram-positive bacteria,
against certain Gram-negative bacteria, in animals including humans,
In vivo by oral and/or parenteral administration
active in vivo ). These in vivo activities are more restricted with respect to the microorganisms being tested, and the activity is determined by infecting mice of substantially uniform body weight with the bacteria to be tested and then administering the test compound orally or by subcutaneous injection. Find out. In practice, for example, 10 mice are inoculated intraperitoneally with a bacterial culture diluted to about 1 to 100 times the LD ₁₀₀ (the lowest concentration of bacteria required for 100% mortality). To check for changes in the virulence of the test bacteria, a baseline test is performed simultaneously using mice inoculated with low dilution bacteria. Test compound was administered 30 minutes after inoculation, 4, 24,
Repeat administration 48 hours later. Four days after administration, surviving mice are retained and the number of survivors is recorded. For in vivo use, these new compounds can be administered orally or parenterally, such as by subcutaneous or intramuscular injection, at approximately 1 kg body weight per day.
Administer mg to 200 mg. A good dosage range is about 5 mg/Kg/day to 100 mg/Kg/day;
Kg/day to 50 mg/Kg/day is suitable. Good excipients for parenteral administration include aqueous vehicles such as water, isotonic saline, isotonic dextrin aqueous Ringer's solution; vegetable oils (cottonseed oil, peanut oil, corn oil, sesame oil), dimethyl sulfonate. other non-aqueous vehicles (glycerin, polyethylene glycol, sorbitol) that do not prevent the therapeutic effect of the formulation and are non-toxic in the amounts or ratios used. Additionally, it may be advantageous to prepare compositions suitable for extemporaneous solutions prepared prior to administration. Such compositions include liquid diluents such as propylene glycol, diethyl carbonate, glycerin, sorbitol; buffers; hyaluronidase;
Local anesthetic; Contains inorganic salts and provides good pharmacological activity. These compounds can be mixed with pharmaceutically suitable inert carriers such as solid diluents, aqueous vehicles, non-toxic organic solvents, and prepared into capsules, tablets, lozenges, troches, dry mixes, suspensions, solutions, etc.
It can be in the form of an elixir, parenteral solution or suspension. Generally the compound will be about 0.5 to 0.5 of the total composition.
Various dosage forms are used at 90 percent concentration. The examples presented below are not written to obtain the highest amount of product produced or to achieve the highest yield of product. The examples are merely illustrative of the steps and products obtained therefrom. In all examples "vanillin/ethanol/H ₃ PO ₄ spray" and "vanillin/H ₃ PO ₄
"Spray" refers to a solution of 1.0 g vanillin, 100 ml ethanol and 100 ml H ₃ PO ₄ . Reference example 1 4″-epi-9-deoxo-9a-(beta-cyanoethyl)-9a-aza-9a-homoerythromycin A 4″-epi-9-deoxo-9a-aza-9a-homoerythromycin A (11.6 g , 15.8 mmol)
Dissolve in 100ml of acrylonitrile. solution 19
Reflux for an hour and concentrate under vacuum to give an ivory foam. TLC check (silica gel board;
Developed with CH ₂ Cl ₂ /CH ₃ OH/concentrated NH ₄ OH = 6/1/0.1; Sprayed with vanillin/H ₃ PO ₄ ; Heated;
Rf0.51) contains trace amounts of impurities (highly polar). Shows a single product (less polar). This is used in the next step without being purified. ¹³ C―nmr [CDCl ₃ , (CH ₃ ) ₄ Si internal standard]
ppm177.92 (lactone C=0), 118.86 (-C≡
N), 102.49 (C-1′), 96.04 (C-1″), 40.25
[(CH ₃ ) ₂ N—]. Reference example 2 4″-epi-9-dexo-9a-(gamma-aminopropyl)-9a-aza-9a-homoerythromycin A 12.8 g dissolved in 250 ml of absolute ethanol
(16.2 mmol) of 4″-epi-9-deoxo-9a
-(beta-cyanoethyl)-9a-other-9a-
A solution of homoerythromycin A is mixed with 12.8 g of Raney Nickel catalyst (50% water-wet) and hydrogenated for 19 hours at 50 psi (3.52 Kg/cm ² ) in a Parr apparatus. TLC check (silica gel plate: CHCl ₃ /
Development with CH ₃ OH/concentrated NH ₄ OH = 6/1/0.1; vanillin/H ₃ PO ₄ spray; heating; Rf 0.13) indicates that the reaction is complete. Filter the catalyst and concentrate the liquid under vacuum to obtain a foam. The crude product is approx.
Dissolve in 100ml of methylene chloride. Shake the solution with an equal volume of saturated aqueous sodium bicarbonate and separate the organic layer. Dry over sodium sulfate and concentrate under vacuum to give an ivory foam (10.5 g). Crystallize with hot ether to obtain 4.0 g. mp135℃ (decomposition) ¹³ C―nmr [CDCl ₃ , (CH ₃ ) ₄ internal standard]
ppm 177.07 (lactone C=O) 102.22 (C-1'),
95.77 (C-1″), 40.30 [(CH ₃ ) ₂ N-]. Example 1 4″-Epi-9,11-deoxo-11beta, 9a-
(epoxypropano)-9a-aza-9a-homoerythromycin A [formula (), n=3, 4″-axial hydroxy] and 4″-epi-9-deoxo-9a-(gamma-acetoxypropyl)-
9a-Aza-9a-Homoerythromycin A 4″-Epi-9- dissolved in 20ml of chloroform
Deoxo-9a- (gamma-aminopropyl)-
9a-Aza-9a-Homoerythromycin A (3.37g,
574 mg (0.66 ml; 4.9 mmol) of amyl nitrite and 511 mg (8.52 mmol) of
of glacial acetic acid. The mixture is refluxed vigorously for 1 hour. TLC check (silica gel plate; briefly immersed in formamide/acetone solution and air-dried.
Developed with CHCl ₃ /acetone = 1/1; Vanillin/
H ₃ PO ₄ spray; heating) indicates completion of the reaction and indicates the presence of two major products. The reaction mixture is shaken with 50 ml of 10% potassium carbonate water and the organic layer is separated. Wash with 50 ml of saturated saline and dry with sodium sulfate. Concentrate under vacuum to give a colorless foam. (3.17g) The formamide-treated silica gel was well stirred.
300g silica gel (230-240 mesh)/600ml
Add 120ml of formamide to the acetone slurry,
A powder is obtained by removing the solvent on a rotary evaporator. Using 300 g of formamide-treated silica gel, the crude product was dissolved in CHCl ₃ /hexane = 98/
The effluent and fractions in step 2 are purified by chromatography checked with TLC (deployment system described above). Two components purified by reaction; the indicated compound (low polarity, 344
mg, 10% yield) and 4″-epi-9-deoxo-9a
-(gamma-acetoxypropyl)-9a-aza-9a-homoerythromycin A (400 mg, 11% yield) is isolated as a colorless amorphous solid. Next, the title product purified by chromatography was crystallized from acetone/water (yield: 94 mg, m.
p.139-141℃) 4″-Epi-9,11-deoxo-11beta,9a-
(Epoxypropano)-9a-aza-9a-homoerythromycin A: ¹ H-nmr (CDCl ₃ ) delta 2.28
[6H, s, (CH ₃ ) ₂ N—], 3.28 (3H, s, cladinose CH ₃ O—]; ¹³ C—nmr [CDCl ₃ , (CH ₃ ) ₄ Si
Internal standard〕ppm176.28 (lactone C=O),
102.36 (C-1′), 96.18 (C-1″), 40.26 [(CH ₃
) ₂ -
N-]; Mass spectrum (m/e) 774 (molecular ion), 616.4, 599.4, 458.3, 442.3, 158.2, 127.1. 4″-epi-9-deoxo-9a-(gamma-acetoxypropyl)-9a-aza-9a-homoerythromycin A: ¹ H-nmr (CDCl ₃ ) delta 2.02
(3H, s, [Formula]), 2.27 (6H, s, (CH ₃ ) ₂ N-), 3.28 (3H, s, Cladinose
_CH3O― ). Reference example 3 9-deoxo-9a- (beta-cyanoethyl)
-9a-aza-9a-homoerythromycin A In 10 ml of acrylonitrile, 9-deoxo-
9a-Aza-9a-Homoerythromycin A (1.0g)
dissolve. The mixture is refluxed for 6 hours. Then stir overnight at room temperature. The mixture is concentrated under vacuum to give a brown foam. Silica gel (40g, 70-230
The crude product was purified by chromatography using a mixed solvent of CH ₂ Cl ₂ /CH ₃ OH/concentrated ammonia = 10/1/0.1, and the fractions were purified by TLC.
(Silica gel plate: developed with CH ₂ Cl ₂ /CH ₃ OH/concentrated ammonia = 6/1/0.1, vanillin/H ₃ PO ₄ spray, heated; Rf 0.57) and checked with colorless foam. Obtain it as a thing. 605 mg (56% yield). ¹ H―nmr (CDCl ₃ ) delta 2.34 [6H, s,
(CH ₃ ) ₂ N―], 3.33 (3H, s, Cladinose
CH ₃ O-); ¹³ C-nmr [CDCl ₃ , (CH ₃ ) ₄ Si internal standard] ppm 177.62 (lactone C=O), 118.85 (-
C≡N), 103.01 (C-1′), 95.91 (C-1″), 40.
33
[(CH ₃ ) ₂ N—]. Reference example 4 9-deoxo-9a-(gamma-aminopropyl)-9a-aza-9a-homoerythromycin A 47 g (59.6 mmol) of 9-deoxo-9a-(beta-cyanoethyl)-9a dissolved in 520 ml of ethanol -Ather-9a- A solution of homoerythromycin A was mixed with 47 g of Raney-nickel catalyst (50% water-wet) at 50 psi in a Parr apparatus.
Hydrogenate for 2.75 hours. In TLC check (Silica gel plate: CHCl ₃ / CH ₃ OH / concentrated NH ₄ OH) 6/
Developed at 1/0.1; Vanillin/H ₃ PO ₄ spray;
heating), indicating that the reaction is not complete at this point.
Add another 25 g of catalyst to the mixture and apply 50 psi.
(3.52Kg/cm ² ) for 1.25 hours. The catalyst is filtered and the liquid is concentrated in vacuo to give a colorless foam. Dissolve the crude product in 600 ml of ethyl acetate.
Stir the solution with 800ml of water and adjust the pH to 9.5 with 6N sodium hydroxide. The organic phase is dried over sodium sulfate and concentrated under vacuum to give a foam. Silica gel chromatography (800g, 70-230 mesh)
Therefore, CHCl ₃ /CH ₃ OH/concentrated NH ₄ OH=6/
Elution at 1/0.05 (Rf 0.15) gives 14.7 g of pure title compound as a colorless foam (31% yield). 1.1
g sample was crystallized with diethyl ether to obtain 545 mg of colorless crystals. mp180−183℃. ¹ H―nmr (CDCl ₃ ) delta 2.30 (6H, s,
(CH ₃ ) ₂ N―], 3.32 (3H, s, cladinose
CH ₃ O-); ¹³ C-nmr [CDCl ₃ , (CH ₃ ) ₄ Si internal standard] ppm 177.01 (lactone C=O), 102.69 (C
-1′), 95.27 (C-1″), 40.33 [(CH ₃ ) ₂ N-). Example 2 9,11-deoxo-11beta, 9a-(epoxypropano)-9a-aza-9a- Homoerythromycin A [formula (), n=3,4″-equatorial hydroxyl] and 9-deoxo-9a-
(gamma-acetoxypropyl)-9a-aza-
9a-Homoerythromycin A 1.01 g (1.16 ml, 8.63 mmol) of amyl nitrite and 0.97 g (0.92 ml, 16.2 mmol) of glacial acetic acid are added and the mixture is refluxed vigorously for 1 hour. The mixture is stirred with 150 ml of water and brought to pH 8.0 with saturated sodium bicarbonate. Separate the organic layer and wash with an equal volume of water. Dry over sodium sulfate and concentrate under vacuum to give a yellow foam. A TLC check shows the presence of two main products in the crude product. (Silica gel plate, 15:85, briefly immersed in formamide/acetone solution. Then air-dried.
Developed with CHCl ₃ /acetone = 1:1, vanillin /
_H3PO4 spray, heat) _. The formamide-treated silica gel was well stirred.
900g silica gel (230-400 mesh)/1800ml
Add 360ml of formamide to the acetone slurry,
The solvent is distilled off using an evaporator to obtain a powder. 900
g of formamide-treated silica gel, initially 2
The crude product (5.8 g) is purified by chromatography, eluting with 1 liter of CHCl ₃ /hexane 7/3 and then 3 liters of CHCl ₃ /hexane 8/2. Check the fractions taken in 5 ml aliquots using TLC (the system described above). In this way, the 2 produced by the reaction
Seed composition - listed compound (low polarity) (0.79g, 13%
yield) and 9-deoxo-9a-(gamma-acetoxypropyl)-9a-aza-9a-homoerythromycin A (0.76 g, 12% yield), respectively, as colorless, amorphous solids. 9,11-deoxo-11beta,9a-(epoxypropano)-9a-aza-9a-homoerythromycin A: ¹ H-nmr (CDCl ₃ ) delta 2.30 [6H,
s, (CH ₃ ) ₂ N―], 3.33 (3H, s, cladinose
CH ₃ O-); ¹³ C-nmr (CDCl ₃ ) ppm 176.3 (lactone C=O), 103.2 (C-1'), 103.2 (C-1')
,
96.2 (C-1″), 40.4 [(CH ₃ ) ₂ N-]; Mass spectrum (m/e) 616, 599, 458, 442, 158, 127. 9-deoxo-9a- (gamma-acetoxypropyl) -9a-Aza-9a-Homoerythromycin A: ¹ H-nmr (CDCl ₃ ) delta 2.01 (3H, s,
[Formula]), 2.29 [3H, s, [Formula]], 3.31 (3H, s, Cladinose CH ₃ O-); Mass spectrum (m/e) 834.6. Reference example 5 9-deoxo-9a-(cyanomethyl)-9a-
Aza-9a-homoerythromycin A3',9a-bis-N-oxide 9-deoxo-9a-hydroxy-9a-aza-9a-homoerythromycin A3'-N-oxide (11.0 g, 14.4
79 g (0.57 mol) of anhydrous potassium carbonate and 69 g (0.58 mol) of bromoacetonitrile are added to the solution. The mixture is stirred at room temperature for 5.25 hours. TLC check (silica gel plate, CHCl ₃ /
Developed with CH ₃ OH/concentrated NH ₄ OH = 6/1/0.1; vanillin/H ₃ PO ₄ spray, heating), the starting material is consumed and the product (low polarity, Rf 0.28) is shown. This indicates the presence of trace amounts of impurities. Strain the mixture;
Concentrate the liquid under vacuum to give a viscous oil. The oil is stirred in 600 ml of diethyl ether to give a granular solid, which is filtered. Wash with 100 ml of ether and air dry. The title compound thus obtained (10.4 g, 89.9% yield, colorless granular solid)
is used as is in the next step. Reference Example 6 9-deoxo-9a-aza-9a-cyanomethyl-9a-homoerythromycin A Place the product described in Reference Example 5 (1.0 g), 10 ml of ethanol, and 0.25 g of 5% Pd-C in a Pearl container. , replace with nitrogen gas repeatedly. Next, 10 psi (0.70 Kg/cm ² ) of hydrogen gas is introduced and hydrogenation is carried out at room temperature for 4.5 hours. The reaction mixture is removed from the vessel, filtered and concentrated under vacuum to give a foam. Dissolve this in 100ml of methylene chloride and add 100ml of water. Add 6N NaOH to adjust pH to 9.5.
Separate the methylene chloride layer and dry (Na ₂ SO ₄ ). Evaporation under vacuum gives 700 mg of foam. Using silica gel (21 g, 70-230 mesh), _CH2Cl2 / _MeOH /concentrated _NH4OH ,
Perform effluent chromatography on 18/1/0.05. Distillation of the fractions yields the title product. (135mg) Rf [silica gel plate; developing system CHCl ₃ /CH ₃ OH/
Concentrated NH ₄ OH = 6/1/0.1; vanillin/H ₃ PO ₄ spray; heating], 0.61 ¹ H−nmr (CDCl ₃ ) delta 2.26 [6H, s,
(CH ₃ ) ₂ N―], 3.28 (3H, s, Cladinose
CH ₃ O-); ¹³ C-nmr [CDCl ₃ , (CH ₃ ) ₄ Si internal standard] ppm 178.0 (lactone C=O), 116.76 (-C
≡N), 102.78 (C-1′), 95.16 (C-1″), 40.30
[(CH ₃ ) ₂ N—]. Reference example 7 9-deoxo-9a-(beta-aminoethyl)
-9a-aza-9a-homoerythromycin A 9-deoxo-9a-cyanomethyl-9a-aza-9a-homoerythromycin A (2.4 g, 3.1 mmol), 72 ml of methanol and anhydrous cobalt chloride (0.792 g, 6.1 mmol) To the mixture is added sodium borohydride (1.17 g, 0.0309 mol) at room temperature. An exothermic reaction occurs and foaming occurs. The mixture is stirred at room temperature for 2 hours and then concentrated in vacuo (aspirator) to give a black oil. Residue 1:1,
Dissolved in 5 ml of water: methylene chloride mixture, 1N
Adjust HClPH to 2.5. Separate the aqueous layer and add 25 ml of methylene chloride. Add 1N NaH and adjust the pH
Make it 9.5. Separate the organic layer and add an equal amount of water.
Add 1N HCl to adjust the pH to 2.0. Separate the aqueous layer again and add 25ml of methylene chloride to 1N
Add NaOH to adjust pH to 9.5. Separate the organic layer;
Dry (Na ₂ SO ₄ ). Concentrate under reduced pressure to obtain the foamy crude product (1.15 g). 1.05 g of the crude product was purified using 30 g of 70-230 mesh silica gel in _CHCl3 : _CH3OH : _NH4OH .
(6:1:0.1) and purified by chromatography in fractions of 5 ml. The target substance (highly polar) is contained in fractions 80-150. These are combined and concentrated to a small volume under reduced pressure. The concentrate is washed with saturated aqueous sodium bicarbonate and dried. Concentration under vacuum gives 75 mg of the title product as a foam. Example 3 9,11-deoxo-11beta,9a-(epoxyethano)-9a-aza-9a-homoerythromycin A [Formula (), n = 2, 4″-equatorial hydroxy] 9-deoxo-9a-( beta-aminoethyl)
-9a-Aza-9a-Homoerythromycin A (0.38
A mixture of chloroform (4 ml), isoamyl nitrite (0.072 ml, 0.536 mmol), and glacial acetic acid (0.056 ml, 0.976 mmol) is refluxed for 1 hour. Additional isoamyl nitrite and glacial acetic acid are added and the mixture is refluxed for 2 hours. Cool and add 10 ml of chloroform and 15 ml of saturated sodium bicarbonate water. Add 1N NaOH to adjust pH to 9.5.
Separate the organic layer and dry (Na ₂ SO ₄ ). Concentrate under reduced pressure to obtain 0.37 g of brown foam. Using 55 g of formamide-treated silica gel (prepared as in Example 1), chloroform:hexane (90:10)
The title product was obtained from fractions 81-130 by column chromatography using the effluent as a fraction of 5 ml. The combined fractions are washed with water and dried (Na ₂ SO ₄ ). Concentrate under reduced pressure to obtain a white solid.
Dissolve the solid in methylene chloride and filter. Evaporation under vacuum gives 26.5 mg of product as a white solid. ¹³ C―nmr [CDCl ₃ , chloroform internal standard]
ppm177.71 (lactone carbonyl), 103.36 (C-
1'), 95.97 (C-1''), 40.40 [(CH ₃ ) ₂ N-]; Mass spectrum (m/e) 602, 444, 428. Example 4 9,11-deoxo-11beta, 9a- (Epoxymethano)-9a-aza-9a-homoerythromycin A [formula (), n=1, 4″-equatorial hydroxy] 10.0 g dissolved in 150 ml of chloroform
(13.64 mmol) of 9-deoxo-9a-aza-9a
-Homoerythromycin A, 1.19g (1.10ml,
37% formaldehyde aqueous solution (14.0 mmol),
A solution of 0.488 g (0.40 ml, 0.01 mol) formic acid is stirred at room temperature (approximately 25° C.) for 70 hours. In TLC (silica gel plate, CH ₂ Cl ₂ /CH ₃ OH/concentrated NH ₄ OH = 6/
Developed at 1/0.1; vanillin/H ₃ PO ₄ spray,
heating) indicates the presence of at least five components. The mixture was stirred with saturated aqueous sodium bicarbonate (400ml) and
Adjust the pH of the aqueous layer to 9.5 with 6N sodium hydroxide. Separate the organic layer and dry with sodium sulfate. Concentrate under vacuum to obtain a colorless foam. The crude product was purified by silica gel chromatography (405 g,
70−230 mesh silica gel; CH ₂ Cl ₂ /
CH ₃ OH/concentrated NH ₄ OH = 12/1/0.05) 563
mg (5.5% yield) of pure title product is obtained as a colorless foam. Rf (silica gel plate; CH ₂ Cl ₂ /
_CH3COH /concentrated _NH4OH =6/1/0.1), 0.5. ¹³ C―nmr [CDCl ₃ , (CH ₃ ) ₄ Si internal standard]
ppm173.37 (lactone C=O), 103.48 (C-1′),
97.07 (C-1″), 83.31 (C-5), 81.01 (-O-
CH ₂ ―N′―), 78.01, 76.11, 75.56, 40.24
[(CH ₃ ) ₂ N-]; Mass spectrum (m/e) 588,
571, 430, 414, 158, 127. Production example A 9-deoxo-9a-aza-9a-hydroxy-
9a-Homoerythromycin A3'-N-oxide 9-deoxo- dissolved in 40 ml of methanol
9a-Aza-9a-Homoerythromycin A (10.0g)
30% hydrogen peroxide solution (50 ml) is added dropwise to the solution under stirring over 5 to 10 minutes. After stirring overnight at room temperature,
The mixture is poured into a stirred slurry of ice (200 g), ethyl acetate (200 ml), water (100 ml).
Decompose excess hydrogen peroxide by carefully adding saturated sodium sulfite water dropwise until the iodine-starch reaction ceases. Separate the layers and wash the aqueous layer twice with 200 ml of ethyl acetate. The organic extracts are combined, dried over sodium sulfate, and evaporated to give crude 9-deoxo-9a-aza-9a-hydroxy-9a-homoerythromycin A3'-N-oxide as a colorless foam (8.6 g). . The crude product is sufficient for use in the following manufacturing process, but purification can be easily carried out by silica gel chromatography (elution with methylene chloride:methanol:concentrated aqueous ammonia system (12:1:0.1)). I can do it. The progress of separation and purification was determined by thin layer chromatography using a silica gel plate (methylene chloride: methanol: concentrated ammonia water, (9:
1:0.1)). The silica gel plate was sprayed with vanillin [ethanol (50ml):
85% H ₃ PO ₄ (50 ml): Vanillin (1.0 g)]
Heat. ¹ H―nmr (CDCl ₃ ) delta 3.21 [6H, s,
[Formula]], 3.39 (3H, s, cladinose CH ₃ O-). MS: Main peak m/e 576 (ion from desource amine fragment), 418 (aglycone ion - both sugars). Both peaks are characteristic of the [formula] residue in the aglycone. Preparation Example B 4″-epi-erythromycin A oxime 4″-epi-erythromycin A (50 g, 0.0646 mol) is dissolved in 265 ml of pyridine. Hydroxylamine hydrochloride (112.2 g, 1.615 mol) is added and the slurry is stirred for 16 hours. The reaction mixture is distilled off to a viscous slurry. Dilute with 300 ml of isopropanol and stir well. 3x after this
Wash with 100 ml isopropanol. The liquid and washing liquid are combined and distilled off to give a water-soluble foam. This is triturated with ether to give the crude product as the hydrochloride salt (100 g). The crude product was dissolved _{in CH2Cl2}
and purified by partitioning into NaHCO ₃ water adjusted to pH 9.5 with dilute NaOH. Separate the aqueous layer and wash with ethyl acetate and then ether. Combine all organic layers and dry (Na ₂ SO ₄ ). This is evaporated to give the title product as a white foam (59.5 g). 59.5g; tlcRf0.5 (60:10: _{1CH2Cl2 :}
CH ₃ OH: concentrated NH ₄ OH); delta 2.31 [6H, s,
(CH ₃ ) ₂ N―], 3.32 (3H, s, cladinose
_CH3O― ). Preparation Example C 4″-Epi-9a-Aza-9a-Homoerythromycin A To a solution of the named product of Preparation B (59.2 g, 0.0787 mol) dissolved in 400 ml of acetone is added 225 ml of
Add a slurry of NaHCO ₃ (60 g) in H ₂ O.
A solution of methanesulyl chloride (36.3 g, 24.5 ml) in 50 ml of acetone is added dropwise over 10 minutes. During this time, keep the temperature below 30°C in a cold bath. The mixture is stirred for 4.5 hours and the acetone is distilled off.
Add CH ₂ Cl ₂ (400 ml) to the aqueous residue and dilute with 6N HCl.
Set the pH to 5.6. Separate the aqueous layer and wash twice with CH ₂ Cl ₂ . Next, adjust the pH to 9.5 with 6N NaOH. The aqueous base solution is extracted with 2x CH ₂ Cl ₂ , 1x ethyl acetate and 1x ether. Combine the extracts and dry (Na ₂ SO ₄ ). This is distilled off to give the title product as an ivory foam. 41g 41g; tlc Rf0.4 (60: ₁₀ : _1CH2Cl2 : _CH3CH :
Concentrated NH ₄ OH); ¹ H―nmr (CDCl ₃ ) delta 2.27
[6H, s, (CH ₃ ) ₂ N—], 3.29 (3H, s, cladinose CH ₃ O—); ¹³ C—nmr [CDCl ₃ , (CH ₃ ) ₄ Si
Internal standard〕ppm177.24 (lactone C=O),
163.53 (amide C=O), 102.29 and 95.24 (C-
3, C-5), 40.22 [(CH ₃ ) ₂ N-]. Production Example D 4″-epi-9-deoxo-9a-aza-9a-homoerythromycin A 600 ml of the product described in Production Example C (40 g)
Dissolve NaBH ₄ in CH ₃ OH while keeping below 38 °C.
(45g) over 45 minutes. The reaction mixture is stirred for 64 hours and evaporated to yield a viscous slurry containing sodium borohydride and the product boron ester complex. Combine the latter complex with 500 ml of CH ₂ Cl ₂
and distribute in 500 ml of water. Repeat the following process three times. Keep the mixture at pH 2.5 with dilute HCl under stirring;
Stir vigorously for 25 minutes; separate the H ₂ O layer and
Combine with ml _{of CH2Cl2} ; bring to PH9.5 with dilute NaOH. Separate the _{two CH2Cl} layers. Combine the _two layers of CH ₂ Cl at PH 9.5 and add 500 ml H ₂ O. After repeating three times, the _two CH ₂ Cl layers were dried (Na ₂ SO ₄ ) and evaporated.
The crude product is obtained as a foam. 34g. 150ml
Crystallize with hot isopropyl ether, cool and dilute with 300 ml of pentane. The pure title product is obtained. 25.8g. White crystal. tlc Rf0.5 (9:1 CHCl ₃ : diethylamine);
Rf0.1 (90:10:1CH ₂ Cl ₂ :CH ₃ OH: concentrated
NH ₄ OH), mp170−180°; ¹ H−nmr (CDCl ₃ ) delta 2.26 [6H, s, (CH ₃ ) ₂ N−], 3.29 (3H, s,
Cladinose _CH3O- ); ^13C -nmr [ _CDCl3 ,
(CH ₃ ) ₄ Si internal standard] ppm 179.44 (Lactone C=
O), 103.57 and 96.70 (C-3, C-5); 41.50
[(CH ₃ ) ₂ -N-]. Production example E 4″-epi-9-deoxo-9a-hydroxy-
9a-Aza-9a-Homoerythromycin A3'-N
- Oxide Stirring, under _N2 gas, the labeled product of Preparation D (3.0
g) in 15 ml of THF:CH ₃ OH, 1:1.
Add 30% H ₂ O ₂ (5 ml). 0.5 hours later, another 30
Add % _H2O2 ( _2.5ml ) and after 0.5h remove excess
Carefully pour the reaction mixture into a 1:1 mixture of CH ₂ Cl ₂ :H ₂ O containing Na ₂ SO ₃ (exothermic). PH is 9. The aqueous layer is washed with CH ₂ Cl ₂ and then with ethyl acetate. Combine the organic layers and dry (Na ₂ SO ₄ ).
Evaporation gives the title product. 2.7g, tlc Rf0.15
(60:10:1, CH ₂ Cl ₂ :CH ₃ OH: concentrated NH ₄ OH):
¹ H-NMR (CDCl ₃ ) delta, 3.21 [6H, s,
(CH ₃ ) ₂ N→O], 3.38 (3H, s, cladinose
MS: Main peak m/e 576 (desousamine fragment ion at C-5 position), 418 ( _N
-Hydroxy aglycone ion-both sugars), both peaks are characteristic of the -N-OH group in the aglycone.

Claims (1)

[Claims] 1 Formula: and pharmaceutically suitable acid addition salts thereof (where n is 1, 2 or 3; the wavy line of the 4''-OH group indicates the axial and equatorial bonds at this position; provided that when n is 1) is a 4″-OH group is an equatorial bond). 2. The compound of claim 1, wherein n is 2. 3. The compound of claim 1, wherein n is 3. 4. The compound according to claim 3, in which the 4"-OH group is an axial bond. 5. The compound according to claim 3, in which the 4"-OH group is an equatorial bond. 6. The compound of claim 1, wherein n is 1. 7. An antibacterial composition comprising a compound of the following formula or a pharmaceutically suitable acid addition salt thereof and a pharmaceutical carrier. (In the formula, n is 1, 2 or 3; the wavy line of the 4''-OH group indicates an axial and equatorial bond at the position; however, when n is 1, the 4''-OH group is an equatorial bond). 8 Formula: of the formula: with at least stoichiometric amounts of isoamyl nitrite and acetic acid in an inert solvent. A method for producing a compound. However, in the above formula, m is 2 or 3; n is 2 or 3; the wavy line in the 4''-OH group represents an axial and equatorial bond at this position. 9 Formula: 9-deoxo-9-aza-9-homoerythromycin A of the formula: A method for producing a compound. 10. The method of claim 9, wherein the solvent is chloroform.