HK1065803B - Process for preparing 4"-substituted-9-deoxo-9a-aza-9a-homoerythromycin a derivatives - Google Patents

Description

Intermediate for preparing 4' -substituted 9-deoxy-9A-aza-9A-homoerythromycin A derivative and preparation method thereof

The present invention is a divisional application of the chinese patent application filed on 25/4/2002 under the name of 02118435.6 entitled "a method for preparing a 4" -substituted 9-deoxo-9A-aza-9A-homoerythromycin a derivative ".

The technical field is as follows:

the present invention relates to a process for the preparation of C-4 "substituted 9-deoxy-9 a-aza-9 a-homoerythromycin a (hereinafter referred to as" azalide ") derivatives useful as antibacterial and antiprotozoal agents in mammals, including humans, as well as fish and birds. The invention also relates to a process for the preparation of a stable intermediate of a target nitrogen impurity in a process for the preparation of the target nitrogen impurity, and to a crystalline salt of the intermediate. The invention also relates to pharmaceutical compositions containing the novel compounds made according to the targeted methods, and methods of treating bacterial and protozoal infections in mammals, fish and birds by administering the novel compounds made according to the targeted methods to mammals, fish and birds in need of such treatment.

Background art:

macrolide antibiotics are known to be effective in the treatment of a broad spectrum of bacterial and protozoal infections in mammals, fish and birds. Such antibiotics include various derivatives of erythromycin A such as the commercially available azithromycin, see U.S. Pat. Nos. 4,474,768 and 4,517,359, both of which are incorporated herein by reference in their entirety. Like azithromycin and other macrolide antibiotics, the macrolide compounds of the present invention have potent activities against various bacterial infections and protozoa infections as described below.

The generation of targeted nitrogen impurities on a commercial scale presents several difficulties, including, but not limited to, lower yields and instability of some synthetic intermediates, as well as the presence of undesirable impurities.

The invention comprises the following contents:

the invention relates to a general formula1A process for the preparation of a compound of (1) or a pharmaceutically acceptable salt thereof,

the method comprises the following steps:

general purpose medicineFormula (II)2The compound of (1):

and formula HNR⁸R¹⁵In an organic solvent comprising isopropanol;

wherein the reaction is carried out at a temperature of at least about 40 ℃;

wherein:

R³is-CH 2NR⁸R¹⁵；

R⁸Is C₁-C₁₀An alkyl group; and is

R¹⁵Is H or C₁-C₁₀An alkyl group.

In a preferred embodiment of the process, R⁸Is propyl and R¹⁵Is H. In a particularly preferred embodiment, R⁸Is n-propyl and R¹⁵Is H.

In a particularly preferred embodiment, the organic solvent is isopropanol.

In another preferred embodiment, the present invention relates to a process of the formula2With n-propylamine in an organic solvent containing isopropanol1a compound of a or a pharmaceutically acceptable salt thereof;

wherein the reaction is carried out at a temperature of at least about 40 ℃. In a particularly preferred embodiment thereof, the organic solvent is isopropanol.

It should be noted that the terms "solution" and "mixture" as used herein, unless otherwise stated, may be used interchangeably regardless of the state of dispersion of the components thereofThe application is as follows. The phrase "organic solvent comprising isopropanol", as used herein, unless otherwise stated, refers to a non-aqueous solvent or a mixture of non-aqueous solvents, wherein at least one solvent is isopropanol. In the present application, the term "formula1The compound of (1) comprises1A compound of formula (II) and1a compound of (a). For all embodiments and preferred embodiments of the process of this application, the general formula1a compound of the formula1Particularly preferred embodiments of the compounds of (1).

In one embodiment of the process described herein, the temperature is less than about 95 deg.C, and in a preferred embodiment of the above process, the temperature is less than about 80 deg.C. In a more preferred embodiment of the above process, the temperature is from about 50 ℃ to about 76 ℃. In a particularly preferred embodiment of the above process, the temperature is from about 50 ℃ to about 55 ℃.

In a preferred embodiment of the process described herein, the reaction is carried out at about atmospheric pressure. In this application, the term "atmospheric pressure" refers to a pressure within the normal meteorological atmospheric pressure range at a particular altitude, while the term "elevated pressure" refers to a pressure above atmospheric pressure. In another embodiment of the process described herein, the reaction is carried out under high pressure. In another embodiment of the invention, triethylamine may be present in addition to isopropanol.

In addition to the applicants' preferred embodiments, the general formula2With amines to produce compounds of the formula1The process of the compound of (1) can be successfully carried out in a solvent not containing isopropanol. The invention therefore also relates to a process for the preparation of a compound of the formula2With a general formula HNR⁸R¹⁵The amine of formula (II) is reacted to prepare1The process of (a), wherein the solvent is selected from the group consisting of benzyl alcohol, acetone, methyl isobutyl ketone, DMSO, t-butanol, n-butanol, diisopropyl ether, a mixture of MTBE and DMF, and combinations thereof, wherein the reaction is carried out at a temperature of at least about 40 ℃. The reaction may be carried out at elevated pressure,preferably at about atmospheric pressure. In a further embodiment of the above process, the reaction is accelerated by the addition of a catalytic amount of a Lewis acid. In one embodiment of the above process, the lewis acid is a reagent such as magnesium bromide, potassium iodide, lithium perchlorate, magnesium perchlorate, lithium tetrafluoroborate, hydrochloride salt of pyridine, or tetrabutylammonium iodide. Preferably, the lewis acid is magnesium bromide.

In one embodiment of the process described herein, the molar amount of amine is at least about that of the formula2Five times the molar amount of the compound of (a). In another embodiment of the process described herein, the concentration of amine in the isopropanol is at least about 5 molar. In a particularly preferred embodiment the concentration of n-propylamine in isopropanol is from about 6 to 7 molar.

In one embodiment of the above process, the general formula2With an amine for at least about 24 hours. In a preferred embodiment of the above process, the amine is present in a molar amount of at least about that of formula2Five times the molar amount of the compound of (1), and the formula2With an amine for at least about 24 hours. In a more preferred embodiment of the above process, the temperature is from about 50 ℃ to about 80 ℃. In a more preferred embodiment of the above process, the molar amount of amine is of the formula2About twenty times the molar amount of the compound of (1), the concentration of the amine in isopropanol is about 6 molar concentration, and the general formula2With an amine at a temperature of from about 50 ℃ to about 55 ℃ for at least about 24 hours.

Another embodiment of the process described herein further comprises the formula1A crystalline free base form of the compound of (a). In one embodiment of the formula1The crystalline free base form of the compound is crystallized from a mixture of aqueous solvents. In a preferred embodiment of the above process, the aqueous solvent mixture comprises water and a non-aqueous solvent selected from the group consisting of methanol, ethanol, isopropanol and acetone. In another embodiment of the formula1The free base form of the compound of (A) is derived from organic (C)₆-C₁₀) An alkane solvent or a mixture of such organic alkane solvents. In a preferred embodiment of the above process, the compounds of formula (I) are1The compound of (a) is crystallized by heating the compound with an alkane solvent followed by cooling. In a preferred embodiment of the above process, the organic (C) is₆-C₁₀) The alkane solvent is selected from heptane or octane, most preferably heptane. In another embodiment, the free base may be derived from the general formula1Acid addition salts of the compounds of (1). It is to be understood that "alkane" as used herein, unless otherwise stated, includes saturated monovalent hydrocarbons having straight, cyclic or branched moieties, or mixtures thereof.

In a further embodiment of the process described herein, the general formula1By treating a solution comprising an acid in a water-miscible solvent1The compound of (1). In a preferred embodiment of the above process, the process comprises the general formula1An acid solution is added to the solution of the compound and water. In a more preferred embodiment of the above process, the acid is phosphoric acid, L-tartaric acid or dibenzoyl-D-tartaric acid. In a particularly preferred embodiment thereof, the acid is phosphoric acid. In another more preferred embodiment of the above process, the solvent comprises ethanol. In another preferred embodiment of the above process, the above process further comprises isolating the compound of formula1Acid addition salts of the compounds.

In one embodiment, the process described herein produces a compound of the formula1The compound of (a) is at least 90% pure, more preferably at least 95% pure, and most preferably at least 98% pure. In particular, the process of the invention produces compounds of the formula1Having the formula1The compounds are of purity for use in the preparation of formulations for parenteral administration. The specification and quality requirements for parenteral formulations are known in the art, for example, the particular purity and particle size of the formulation in solution for sterility and pyrogen removal (see Remington's Pharmaceutical Sciences, Mack publishing, Inc.)ing Company, Easton, Pa., eighteenth edition, Gennaro, ed. (1990), p 1545-1580.

In another more preferred embodiment of the above process, the above process further comprises treating the compound of formula (la) with a base in a mixture of water and a non-polar solvent1Acid addition salts of compounds to give compounds of the general formula1The free base form of the compound. In a more preferred embodiment of the above process, the base is a binary carbonate, and in a particularly preferred embodiment, the binary carbonate is potassium carbonate. In another more preferred embodiment thereof the non-polar solvent is dichloromethane. In another preferred embodiment, the process further comprises crystallizing the compound of formula (la) as described above1The free base form of the compound and wherein reference is made to another embodiment as described above.

The invention also relates to a general formula2A process for preparing a compound of (1), comprising:

(a) general formula (II)3In the form of the free base of the compound of (1)

Reacting with sulfonium methyl ylide (sulfonium methyl iodide) ions;

(b) terminating the reaction of step (a) with an aqueous weak acid and partitioning the product in a non-aqueous solution; and is

(c) Deprotecting the product of step (b) to give a compound of formula 2;

wherein R is⁴Is a hydroxyl protecting group.

In one embodiment, the above process further comprises the general formula2Isolation of the compound of (1). In a preferred embodiment of the above process, the compounds of formula (I) are2Is isolated as a hydrate, more preferably as a monohydrate. In the embodiments of the above-mentioned methodThe water content is determined by the Karl Fischer method. In one embodiment of the above process, the hydrate is formed from a compound containing formula2In a mixture with a solvent or a solvent mixture selected from the group consisting of acetone, acetone/water, acetone/heptane and MTBE/heptane. In other embodiments, of formula (III)2Is isolated in the form of its acetate, L-tartrate or dibenzoyl-D-tartrate salt.

The invention relates to a compound of the general formula2A monohydrate of the compound. In a preferred embodiment of the above process, R⁴Is benzyloxycarbonyl.

In another preferred embodiment of the above process, step (a) is carried out at a temperature of from about-80 ℃ to about-45 ℃.

In another embodiment of the above process, the compounds of formula (II) are3The free base of the compound can be prepared from an acid addition salt of the compound of formula 3. In a preferred embodiment of the above process, the acid addition salt is a trifluoroacetic acid addition salt. In another embodiment of the above process, the compounds of formula (II) are3The acid addition salt of the compound is selected from dibenzoyl-D-tartrate, L-tartrate, or phosphate. The acid addition salts of the compounds disclosed herein can be readily prepared by conventional methods.

In one embodiment of the above process, the sulfonium methylylide is dimethylsulfonium methylylide. In a preferred embodiment of the above process dimethylsulfonium methylylide is prepared by reacting trimethylsulfonium halide or sulfonate with a strong base. In a more preferred embodiment of the above process, trimethylsulfonium halide, preferably trimethylsulfonium bromide, is used. In another more preferred embodiment of the above process, trimethylsulfonium halide is reacted with a strong base in an inert organic solvent or a mixture thereof. In another particularly preferred embodiment of the above process, the inert organic solvent is an ethereal solvent, most preferably tetrahydrofuran, or a mixture of tetrahydrofuran and dichloromethane.

In one embodiment, step (c) comprises catalytic hydrogenation, wherein R is⁴Is benzyloxycarbonyl. In a preferred embodiment of the above process, the catalyst used for the hydrogenation is a palladium on carbon catalyst. In a particularly preferred embodiment, the palladium on carbon catalyst is 10% Pd/C (Johnson-Matthey type A402028-10). In a further embodiment of step (C), the product of step (b) is deprotected by catalytic transfer hydrogenation, preferably with ammonium formate, Pd/C in methanol. In a further embodiment, the product of step (b) is treated with bleaching earth prior to hydrogenation. Suitable solvents for the hydrogenation process are acetone, ethyl acetate, THF, MTBE, isopropanol, ethanol and methanol. The preferred solvent is acetone.

The invention also relates to 2' -benzyloxycarbonyl protected compounds II:

this compound is obtained by omitting step (c) of the above-mentioned method.

The invention relates to a general formula3The preparation method of the compound (2):

by oxidation of the general formula4C-4' hydroxy preparation of Compounds

Wherein R is⁴Is a hydroxyl protecting group.

In one embodiment, the oxidation is carried out by reacting a compound comprising the general formula4Adding dimethyl sulfoxide ("DMSO") to a solution of the compound and solvent, cooling the mixture to about-70 deg.C, and adding trifluoroacetic acidAnhydride, followed by addition of triethylamine. In another embodiment, DMSO employs oxalyl chloride (with or without trimethylsilyl acetamide present), polyphosphoric acid, pyridine, SO₃Or acetic anhydride activation. In a further embodiment of the above process, the temperature is maintained between-70 ℃ and-60 ℃ during the addition of trifluoroacetic anhydride. In another preferred embodiment of the above process the solvent is dichloromethane. A particular advantage of the above method is the in situ activation of DMSO in the presence of the reacting alcohol, which avoids the formation of impurities typically encountered in the oxidation of activated DMSO, which typically involves the introduction of an alcohol into a solution containing activated DMSO.

In one embodiment, the above method further comprises the general formula3Isolation of the acid addition salt of the compound. In a preferred embodiment, the acid addition salt is a dibenzoyl-D-tartrate salt or a phosphate salt. In a particularly preferred embodiment, the present invention relates to a process for the preparation of compounds of the general formula3A process for the preparation of an acid addition salt of a compound of formula (I), which process comprises treating a compound of formula (II) with trifluoroacetic acid3A compound of (1); crystallizing the resulting acid addition salt;

wherein R is⁴Is a hydroxyl protecting group.

In a preferred embodiment of the above process, R⁴Is benzyloxycarbonyl.

In another preferred embodiment of the above process, the acid addition salt is crystallized from isopropanol.

In another preferred embodiment of the above process, the acid addition salt is crystallized from a mixture of dichloromethane and methyl tert-butyl ether.

The trifluoroacetic acid addition salts prepared by the process of the invention are not pharmaceutically acceptable, but provide excellent purification and stability, in the general formula1The compounds can be stored and transported as suitable starting materials in the industrial preparation.

In one embodiment of the above process, the general formula4Of (2) to (b)The compound is represented by the protection general formula5The 2' -hydroxyl group of the compound is prepared,

in a preferred embodiment, the 2' -hydroxy group is protected with a benzyloxycarbonyl group. In another preferred embodiment, of formula (III)5Is reacted with at least two molar equivalents of benzylformyl chloride. In a more preferred embodiment, the reaction is carried out in dichloromethane. In a more preferred embodiment, the dichloromethane is present in a volume excess of at least 15 times the volume of the starting material. The invention also relates to the general formula3Trifluoroacetic acid addition salts of compounds wherein R⁴Is benzyloxycarbonyl:

in a preferred embodiment of the above process, the salt has the formula3a structure shown in the specification:

wherein R is⁴Is benzyloxycarbonyl.

The invention also relates to the general formula3dibenzoyl-D-tartrate salt of compound, wherein R⁴Is benzyloxycarbonyl:

the term "hydroxy-protecting group" as used herein, unless otherwise indicated, includes acetyl, benzyloxycarbonyl, and various hydroxy-protecting Groups well known to those skilled In the art, including T.W.Greene, P.G.M.Wuts, In "Protective Groups In organic Synthesis" (J.Wiley&Sons, 1991). Preferably, the hydroxy-protecting group R⁴Is benzyloxycarbonyl ("CBZ").

The term "halogen", as used herein, unless otherwise indicated, includes fluorine, chlorine, or bromine, and the term "halide" refers to the corresponding monovalent anion, which is F-, Cl-, or Br-, respectively.

As used herein, "alkyl", unless otherwise indicated, includes saturated monovalent hydrocarbon radicals having straight, cyclic or branched moieties, or mixtures thereof.

The phrase "pharmaceutically acceptable salts" as used herein, unless otherwise indicated, includes salts of acidic or basic groups of the compounds of the present invention. The compounds prepared by the process of the invention are basic in nature, such as, in particular, of the formula1The free base forms of the compounds are capable of forming a wide variety of salts with a variety of inorganic and organic acids. Acids which may be used to prepare pharmaceutically acceptable acid addition salts of such basic compounds of the present invention are those which form non-toxic acid addition salts, i.e. salts containing pharmacologically acceptable anions, such as hydrochloride, hydrobromide, hydroiodide, nitrate, sulphate, bisulfate, phosphate, acid phosphate, isonicotinate, acetate, lactate, salicylate, citrate, acid citrate, tartrate, pantothenate, bitartrate, ascorbic acid, succinate, maleate, gentisate, fumarate, gluconate, glucarate, saccharate, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate and pamoate [ i.e., 1, 1' -methylene-bis- (2-hydroxy-3-naphthoate) ].]. The compounds prepared by the process of the present invention include an amino moiety, and may form pharmaceutically acceptable salts with various amino acids other than the acids described above.

The term "treatment" as used herein, unless otherwise indicated, includes the treatment or prevention of a bacterial infection or a protozoal infection provided by the methods of the present invention.

The present invention includes compounds of the present invention and their pharmaceutically acceptable salts, wherein one or more of their hydrogen, carbon, nitrogen or other atoms may be replaced by their isotopes. Such compounds can be used as research and diagnostic tools for pharmacokinetic studies and for binding assays.

Detailed description of the invention

The process of the present invention can be carried out according to the following schemes 1 to 4 and the following description. In the following schemes, the substituents R unless otherwise stated³，R⁴，R⁸And R¹⁵As defined above.

Reaction scheme1

Reaction scheme2

Reaction scheme3

General formula for the starting materials for the process of the invention4From which R can be easily derived⁴Preparation of compound 5 as hydrogen is described in WO98/56802, and U.S. Pat. Nos. 4,328,334, 4,474,768, and 4,517,359, all of which are incorporated herein by reference in their entirety.

The reaction schemes given above are merely illustrative and are further detailed below and in the examples below. In scheme 1, the general formula2To the epoxide of the formula1Wherein R is³is-CH₂NR¹⁵R⁸Wherein R is¹⁵And R⁸As defined above. In the most preferred embodiment of the invention, the amine is n-propylamine, i.e., R⁸Is n-propyl and R¹⁵Is H.

To prepare a general formula1Preferably in the presence of a suitable solvent such as isopropanol, or a mixture of organic solvents comprising isopropanol, preferably at a temperature of from about 40 ℃ to about 95 ℃2With a compound of the formula HNR¹⁵R⁸Treatment of a compound of (1), wherein R¹⁵And R⁸As defined above. The optimum temperature for carrying out the reaction is from about 50 ℃ to about 55 ℃, although higher temperatures, e.g., 76 ℃, may also be used. The most preferred pressure at which the reaction is carried out is at about atmospheric pressure; however, the reaction can also be carried out under elevated pressure.

In a previously open formula2Protection of the 2' -hydroxy group in the epoxide Process (see WO98/56802, examples 48, 50, 51 and 70) and the general formula1Production of the Compound (or formula)1a, respectively) requires simultaneous hydrolysis of the protecting group and amination of the epoxide. This method is undesirable because hydrolysis during the epoxide opening step is inefficient and the general formula is subject to the presence of unhydrolyzed protecting groups and other impurities1Isolation of the compounds is more difficult. In another prior process, formula2The compound of (2' -hydroxy is unprotected), is reacted with pure alkylamine, i.e. in the absence of an organic solvent. In this case, the reaction proceeds slowly at the normal boiling temperature of n-propylamine (about 48 ℃ C.). Therefore, in order to generate an elevated temperature, the reaction has to be carried out at high pressure, a feature which is less desirable on an industrial scale. (see WO98/56802, example 8 (preparation 2), 11% yield). In addition, a catalyst is used in the reaction. Applicants have found that a mixture of n-propylamine and isopropyl alcohol having a boiling point of 76 ℃ at laboratory atmospheric pressure allows the reaction to be carried out at temperatures of about 50 ℃ to 55 ℃ in high yields (greater than 85%) without the use of pressurized reaction vessels or catalysts. Applicants' method provides more than previous methodsHigh yield (85%) and better purity, and various crystallization methods can be used for the free base form and the general formula1Acid salts of compounds to give the general formula1Highly pure forms of the compounds, for example, may be used in formulations for which parenteral administration is desired.

In scheme 2, the general formula2Can be prepared by treating a compound of formula (II) with a sulfonium methyl ylide at a temperature of from about-80 ℃ to about-45 ℃3Followed by removal of the 2' -protecting group by conventional methods to provide compounds of formula (II a)2The compound of (1). The starting materials for the process of scheme 2 are preferably of the formula3Trifluoroacetic acid addition salts of the compounds, which are first converted to the free base form, cooled to a low temperature, about-70 ℃, and then reacted with a low temperature solution of the thiomethylate. The sulfomethylate is preferably dimethylsulfonium methylylide, e.g., (CH)₃)₂S+CH₂Prepared by conventional methods, e.g. in ethereal solvents, e.g. THF, or in CH₂CI₂DMF, or DMSO, or mixtures of two or more of the above solvents, by treating the trimethylsulfonium salt, e.g., (CH), with an active agent such as potassium hydroxide, potassium tert-butoxide, sodium tert-butoxide, potassium ethoxide, sodium ethoxide, potassium hexamethyldisilazide (KHMDS) or sodium methoxide, preferably potassium tert-butoxide₃)₃SX, wherein X is halogen, preferably bromine, or sulfonate, more preferably trimethylsulfonium bromide. The protecting group is removed by conventional means, e.g. when R is⁴CBZ is removed by catalytic hydrogenation.

In scheme 3, the 4' ketone is prepared in a continuous process from the general formula5The preparation of the compound of (1). In the first step of the process, the 2' hydroxyl group is selectively protected by conventional means, preferably by treatment with a solution of benzylformyl chloride in dichloromethane wherein R is⁴Is of the formula52' -hydroxy of the compound to give R⁴Is the general formula of benzyloxycarbonyl ("CBZ")4The compound of (1). Preferably at least 2 molar equivalents of benzylformyl chloride are used to ensure 2'Complete conversion of the hydroxyl group to its protected form. Preferably, dichloromethane is used as the solvent to carry out the reaction therein using at least 15 volumes of dichloromethane relative to the volume of the starting material to minimize the formation of bis-CBZ impurities. General formula (VII)4Wherein R is⁴For CBZ, isolated as its dibenzoyl-D-tartrate salt, this method scavenges potential bis-benzyloxycarbonyl impurities. However, due to the general formula4The presence of benzylamine formed by the alkylation of a compound with an amine of benzyl chloride (formed by the decomposition of benzylcarbonyl chloride) destabilizes the isolated product and thus the general formula4The water extraction treatment of the compound of (4) is not preferable. Therefore, the general formula (VII) is preferably not isolated after the protection step of the reaction mixture4The compound (2) is directly transferred to the second step. The second step, which may be carried out in the same vessel as the first step, comprises the oxidation of the 4 "-hydroxyl group to give the general formula34 "-ketone of (1). The oxidation is preferably an activated-DMSO oxidation as described above, i.e. an in situ activation of DMSO performed at reduced temperature, e.g. -60 to-70 ℃, and comprising the addition of trifluoroacetic anhydride followed by triethylamine to a cooled DMSO solution of the compound. Then water was added to the reaction mixture and gradually warmed to room temperature. The mixture is preferably washed in water to give a mixture of formula3Solutions of the compounds.

General formula (VII)3The trifluoroacetate salt of compound (ii) can be prepared by washing the reaction mixture of the oxidation step with water, followed by the addition of trifluoroacetic acid, followed by the addition of a solvent suitable for crystallization of the salt, such as isopropanol or a mixture of dichloromethane and methyl tert-butyl ether ("MTBE"). Other acid addition salts, such as dibenzoyl-D-tartrate and phosphate, may also be prepared in conventional manner. dibenzoyl-D-tartrate and phosphate salts are effective for use in the process of the invention, but are less preferred than trifluoroacetic acid.

As shown in scheme 4, the present invention relates generally to a two-step process for the preparation of compounds of the general formula1Methods of Compounds: in the first step, in a single containerIn-situ preparation general formula3A process for preparing a compound of the formula5Benzyloxycarbonyl protection of the 2' -hydroxy group of the compound of (1) to give the compound of the formula4Followed by direct reaction of a compound of the formula4Oxidation of the 4 "-hydroxyl group of (a) to give a compound of the formula3Preferably as the trifluoroacetic acid addition salt thereof. In the second step, the compound of formula3The free base form of the compound of (a) is preferably prepared from its trifluoroacetate salt, is converted to the compound of formula (la)2Removing the 2 '-protecting group to recover the 2' -hydroxyl group, which epoxide is ring-opened with an amine by heating in a mixture containing isopropanol to give a compound of the general formula1The compound of (1).

Reaction scheme4

First step and second step

The compounds prepared by the process of the present invention are basic in nature and are capable of forming a wide variety of different salts with a variety of inorganic and organic acids. Although such salts must be pharmaceutically acceptable for mammalian administration, it is common practice to initially isolate a compound which is a pharmaceutically unacceptable salt prepared from the reaction mixture by the process of the present invention, and then simply convert the latter to the free base compound by treatment with a basic agent for subsequent reaction or for the preparation of a pharmaceutically acceptable acid addition salt. The acid addition salts of the basic compounds prepared by the process of the present invention can be readily prepared by treating the basic compound with substantially equivalent amounts of the selected inorganic or organic acid in an aqueous solvent medium or a suitable organic solvent. Careful evaporation of the solvent readily affords the desired solid salt. As requiredSalts may also be precipitated from solutions of the free base in an organic solvent by addition of a suitable inorganic or organic acid to the solution. The general formula prepared by the invention1The compounds of (a) and their pharmaceutically acceptable salts (hereinafter referred to as "active compounds") may be administered by the oral, parenteral, topical, or rectal route in the treatment of bacterial and protozoal infections.

In general, the active compound is preferably administered in a dosage range of from about 0.2 milligrams per kg body weight per day (mg/kg/day) to about 200 mg/kg/day, in single or divided doses (i.e., from 1 to 4 doses per day), although these will necessarily vary with the species, body weight and condition of the subject being treated and the particular route of administration chosen. However, it is most preferred to use dosage levels of from about 4 mg/kg/day to about 50 mg/kg/day. However, will vary depending upon the species of mammal, fish or bird being treated, and the individual response to the agent, as well as the type of pharmaceutical formulation selected, and the time period and interval over which such administration is carried out. In some instances, dosage levels below the lower limit of the aforesaid range are sufficient, while in other instances no harmful side effects are caused despite the use of large doses, provided that such large doses are first divided into small doses for administration throughout the day.

The active compounds may be administered alone or in combination with a pharmaceutically acceptable carrier or diluent by the routes previously described, and such administration may be in single or multiple doses. More particularly, the active compounds may be administered in a wide variety of different dosage forms, i.e., they may be combined with various pharmaceutically acceptable inert carriers in the form of tablets, capsules, lozenges, troches, hard gelatin capsules, powders, sprays, emulsions, salves, suppositories, gels, ointments, lotions, ointments, aqueous suspensions, injectable solutions, elixirs, syrups, and the like. Such carriers include solid diluents or fillers, sterile aqueous media and various non-toxic organic solvents and the like. Furthermore, oral pharmaceutical compositions may suitably be sweetened and/or flavoured. Typically, the active compound is present in such dosage forms at a concentration ranging from about 5.0% to about 70% by weight.

For oral administration, tablets containing various excipients such as microcrystalline cellulose, sodium citrate, calcium carbonate, dicalcium phosphate and glycine may be employed along with various disintegrants such as starch (and preferably corn, potato or tapioca starch), alginic acid and certain complex silicates, together with granulation binders such as polyvinylpyrrolidone, sucrose, gelatin and acacia. Additionally, lubricating agents such as magnesium stearate, sodium lauryl sulfate, and talc are often useful for tableting. Solid compositions of the same type may be used as fillers in capsules; preferred materials in this regard also include lactose or milk sugar and high molecular weight polyethylene glycols. When aqueous suspensions and/or elixirs are desired for oral use, the active compound may be combined with various sweetening or flavouring agents, colouring matter or dyes and, if desired, emulsifying and/or suspending agents, together with diluents such as water, ethanol, propylene glycol, glycerin and similar combinations of each.

For parenteral administration, solutions of the active compound can be used either as sesame or peanut oil or as aqueous propylene glycol. The aqueous solution should be suitably buffered if necessary and the liquid diluent first gives isotonic compensation. These aqueous solutions are suitable for intravenous purposes. The oily solutions are suitable for intra-articular, intramuscular and subcutaneous injection purposes. The preparation of all these solutions under sterile conditions can be readily accomplished by standard pharmaceutical techniques known to those skilled in the art.

In addition, the active compounds of the present invention may also be administered topically, by the route of creams, gels, ointments, patches, ointments and the like, in accordance with standard pharmaceutical practice.

For administration to animals other than humans, such as cattle or livestock, the active compound may be administered orally in an animal feed or veterinary drench composition.

The active compounds may also be administered in the form of liposomes, such as small unilamellar liposomes, large unilamellar liposomes and multilamellar liposome delivery systems. Liposomes can be composed of various phospholipids, such as cholesterol, stearylamine or phosphatidylcholines.

The active compound may also be conjugated to a soluble polymer as a carrier for the drug of interest. Such polymers may include polyvinylpyrrolidone, pyran copolymer, polyhydroxypropylmethacrylamide phenyl, polyhydroxyethylaspartamide-phenol, or polyethylene oxide-polylysine substituted with palmitoyl residues. In addition, the active compounds may be used with biodegradable polymer classes effective for achieving controlled release of drugs, for example, polyacids, polyglycolic acids, copolymers of polyacids and polyglycolic acids, polyepsilon caprolactones, polyhydroxybutyric acids, polyorthoesters, polyacetals, polydihydropyrans, polycyanoacrylates and crosslinked or hydrogel amphiphilic group-blocked copolymers.

Detailed Description

The following examples further illustrate the processes and intermediates of this invention. It should be understood that the invention is not limited to the specific details provided in the following examples.

Example 1

Preparation of (2R, 3S, 4R, 5R, 8R, 10R, 11R, 12S, 13S, 14R) -13- [ (2, 6-dideoxy-3-C-methyl-3-O-methyl- α -L-ribohexopyranosyl) oxy ] -2-ethyl-3, 4, 10-trihydryl-3, 5, 8, 10, 12, 14-hexamethyl-11- [ [3, 4, 6-trideoxy-3- (dimethylamino) -2-O- [ (phenylmethoxy) carbonyl ] - β -D-xylopyranosyl ] oxy ] -1-oxa-6-azacyclopentadecan-15-one

To a solution of 25kg of (2R, 3S, 4R, 5R, 8R, 10R, 11R, 12S, 13S, 14R) -13- [ (2, 6-dideoxy-3-C-methyl-3-O-methyl-. alpha. -L-ribopyranosyl) oxy ] -2-ethyl-3, 4, 10-trihydroxy-3, 5, 8, 10, 12, 14-hexamethyl-11- [3, 4, 6-trideoxy-3- (dimethylamino) -beta-D-xylopyranosyl ] oxy ] -1-oxa-6-azacyclopentadecane-15-one in 425L of dichloromethane was added a solution of 13.7kg of benzylformyl chloride cooled to 0-5 ℃ at a rate of keeping the temperature below 5 ℃. The resulting mixture was stirred at this temperature for three hours and then concentrated to 148 liters to give a dry solution containing approximately 26.6kg (90%) of the product (by HPLC-Waters Symmetry C8, 15 cm. times.3.9 mM internal diameter column, 25mM potassium phosphate buffer (pH 7.5) acetonitrile: methanol (35: 50: 15) mobile phase, 2.0ml/mM flow rate, electrochemical detection, retention time 8.2 minutes). This mixture was used directly in example 2.

Example 2

Preparation of (2R, 3S, 4R, 5R, 8R, 10R, 11R, 12S, 13S, 14R) -13- [ (2, 6-dideoxy-3-C-methyl-3-O-methyl- α -L-ribopyranosyl) oxy ] -2-ethyl-3, 4, 10-trihydroxy-3, 5, 8, 10, 12, 14-hexamethyl-11- [ [3, 4, 6-trideoxy-3- (dimethylamino) -2-O- [ (phenylmethoxy) carbonyl ] - β -D-xylopyranosyl ] oxy ] -1-oxa-6-azacyclopentadecan-15-one bistrifluoroacetate

To the solution obtained in example 1 was added 58.6kg of dimethyl sulfoxide ("DMSO"), followed by cooling to-70 ℃. The temperature was maintained between-70 and-60 ℃, 16kg of trifluoroacetic anhydride was added, the mixture was then stirred for 30 minutes, 17.2kg of triethylamine was then added, and the resulting mixture was stirred for a further 30 minutes. The reaction mixture was added to 175 liters of water, gradually warmed to ambient temperature and then the layers were separated. The organic layer was washed twice with 170 liters of water and then concentrated to about 100 liters. Next, 7.8kg of trifluoroacetic acid was added followed by 236 liters of isopropanol, and the mixture was concentrated to crystallize 29.5kg (87.9%) of product (98% purity by HPLC) therefrom.

Analyzing data: mp 187-. Elemental analysis (C)₄₉H₇₆F₆N₂O₁₈Theoretical value: c, 53.74; h, 6.99; f, 10.41; n, 2.56; measured value: c, 53.87; h, 6.99; f, 10.12; and N, 2.59. HPLC system: same as example 1; retention time 9.5 min X-ray powder diffraction (d space): 6.3,8.3,8.8,9.4，10.8，11.8，12.6，13.0，14.3，15.4，15.9，16.4，17.1，17.4，17.8，18.1，19.1，19.8，20.4，21.1，21.5，21.7，22.8，23.4，24.0。

Example 3

Preparation of (2R, 3S, 4R, 5R, 8R, 10R, 11R, 12S, 13S, 14R) -2-ethyl-3, 4, 10-trihydroxy-13- [ [ (3S, 4S, 6R, 8R) -8-methoxy-4, 8-dimethyl-1, 5-dioxaspiro [2.5] oct-6-yl ] oxy ]3, 5, 8, 10, 12, 14-hexamethyl-11- [ [3, 4, 6-trideoxy-3- (dimethylamino) -2-O- [ (phenylmethoxy) carbonyl ] -beta-D-hexylglucopyranosyl ] oxy ] -1-oxa-6-azacyclopentadecan-15-one

(a) 109kg of the product of example 2 in 327 l of dichloromethane were treated with 27.5kg of potassium carbonate in 327 l of water. The layers were separated, the aqueous layer was washed with 327 liters of dichloromethane, the combined organic layers were dried and evaporated to about 327 liters and cooled to-70 ℃.

(b) In a separate vessel, a suspension of 29.7kg trimethylsulfonium bromide in 436 liters of tetrahydrofuran ("THF") was evaporated to about 170 liters, cooled to-12 deg.C, and treated with 36.8kg of potassium tert-butoxide at-10 to-15 deg.C for 75 minutes. The mixture is then added to the dichloromethane solution of step (a) over about 30 minutes, maintaining the temperature at-70 to-80 ℃, and the resulting mixture is then warmed to-65 ℃ and stirred for at least 1 hour. This mixture was then added to 469 l of an aqueous solution of 55.4kg of ammonium chloride. After stirring the mixture at 15-25 ℃ for 15 minutes, the layers were separated, the aqueous layer was washed with 360 l dichloromethane and the combined organic layers were evaporated to approximately 227 l. To the resulting mixture was added 750 liters of acetone. The mixture was finally evaporated to 227 l of a solution containing approximately 70.1kg (80%) of the title product (HPLC system: Metasil AQ C18 column (from MetaChem, batch No. 0520-. This mixture was used directly in example 4.

Example 4

Preparation of (2R, 3S, 4R, 5R, 8R, 10R, 11R, 12S, 13S, 14R) -2-ethyl-3, 4, 10-trihydroxy-13- [ [ (3S, 4S, 6R, 8R) -8-methoxy-4, 8-dimethyl-1, 5-dioxaspiro [2.5] oct-6-yl ] oxy ]3, 5, 8, 10, 12, 14-hexamethyl-11- [ [3, 4, 6-trideoxy-3- (dimethylamino) - β -D-hexylglucopyranosyl ] oxy ] -1-oxa-6-azacyclopentadecan-15-one

The solution containing the product of example 3 was mixed with 11kg of activated carbon, 17.5kg of 10% palladium on carbon (Johnson-Matthey type A402O28-10) and 637 liters of acetone. The resulting mixture was treated with hydrogen at 20-25 deg.C under a pressure of 50psi until the reaction was complete, and then filtered. The filtrate was concentrated to about 350 liters, followed by the addition of 1055 liters of water over 90 minutes. The crystalline product was collected by filtration, washed with a mixture of 132 l of water and 45 l of acetone and dried to yield 57.5kg (94.4%) of the monohydrate of the title epoxide (water content determined by the Kaplan method).

Analyzing data: HPLC system: same as in example 3; retention time 13.3 min. X-ray powder diffraction (d space): 6.0,8.5,9.4, 11.9, 12.7, 13.4, 15.2, 16.9, 17.5, 18.0, 18.9, 19.4, 19.9, 20.7, 21.2, 21.6, 22.8.

Example 5

Preparation of (2R, 3S, 4R, 5R, 8R, 10R, 11R, 12S, 13S, 14R) -13- [ (2, 6-dideoxy-3-C-methyl-3-O-methyl-4-C- [ (propylamino) methyl ] α -L-hexopyranosyl) oxy ] -2-ethyl-3, 4, 10-trihydroxy-3, 5, 8, 10, 12, 14-hexamethyl-11- [ [3, 4, 6-trideoxy-3- (dimethylamino) - β -D-hexopyranosyl ] oxy ] -1-oxa-6-azacyclopentadecan-15-one bisphosphate

56kg of the epoxide monohydrate of example 4 were mixed with 280 liters of isopropanol and 108.2kg of n-propylamine. The mixture was heated at 50-55 ℃ for thirty hours and then concentrated under vacuum to about 112 liters. To this concentrate was added 560 liters of ethanol and 44.8 liters of water. To the resulting mixture was added over a period of about two hours a solution of 16.8kg phosphoric acid in 252 liters of ethanol and crystallized to give the product. After stirring the resulting suspension for 18 hours, the mixture was filtered, the solid was washed with 28 liters of ethanol, and the product was dried to obtain 64.6kg (88%) of the title compound (by HPLC-HPLC system: YMC-Pack Pro C18(YMC Inc. part # AS-12S03-1546WT), 50mM dipotassium hydrogen phosphate buffer (pH 8.0): acetonitrile: methanol 61: 21: 18 mobile phase, 1.0ml/mM flow rate, electrochemical detection. retention time: 26.4 minutes).

Example 6

Preparation of (2R, 3S, 4R, 5R, 8R, 10R, 11R, 12S, 13S, 14R) -13- [ (2, 6-dideoxy-3-C-methyl-3-O-methyl-4-C- [ (propylamino) methyl ] - α -L-hexopyranosyl) oxy-2-ethyl-3, 4, 10-trihydroxy-3, 5, 8, 10, 12, 14-hexamethyl-11- [ [3, 4, 6-trideoxy-3- (dimethylamino) - β -D-hexopyranosyl ] oxy ] -1-oxa-6-azacyclopentadecan-15-one free base

64.6kg of the product from example 5 were mixed with 433 l of dichloromethane, 433 l of water and 27.6kg of potassium carbonate. After stirring the mixture for thirty minutes, the layers were separated and the aqueous layer was washed with 32 liters of dichloromethane. The combined organic layers were clarified by filtration and then evaporated to about 155 liters. To this concentrate 386 liters of heptane were added and the solution was evaporated to about 155 liters and then cooled to 20-25 ℃ for crystallization. After stirring the resulting suspension for six hours, the solid was collected by filtration, washed with 110 l of heptane and dried to yield 40.3kg (77%) of the title compound (by HPLC; same HPLC system as in example 5; retention time 26.4 minutes).

Claims (5)

1. Preparation general formula2The method of (a) to (b),

the method comprises the following steps:

(a) general formula (II)3The free base form of the compound of (a):

reacting with sulfonium methyl ylide ions;

(b) terminating the reaction of step (a) with an aqueous weak acid and partitioning the product in a non-aqueous solution; and is

(c) Deprotection of the product of step (b) to give compounds of the formula2A compound of (1);

wherein R is⁴Is a hydroxy protecting group;

wherein the general formula3The free base form of the compound is prepared from the trifluoroacetic acid addition salt of the compound of formula 3.

2. The method of claim 1, wherein R⁴Is benzyloxycarbonyl.

3. The process of claim 2, wherein step (c) comprises catalytic hydrogenation.

4. The method of claim 1, wherein the compound of formula 2 is a monohydrate.

5. The process of any of claims 1-4 wherein3The reaction of the compound of (a) with a sulfonium methylylide ion is carried out at a temperature of-80 ℃ to-45 ℃.