CZ2000224A3

CZ2000224A3 - Alatrofloxacin parenteral composition

Info

Publication number: CZ2000224A3
Application number: CZ2000224A
Authority: CZ
Inventors: Robert Mark Guinn; John Francis Lambert; Subramanian Sam Guhan; Stanley Walter Walinsky
Original assignee: Pfizer Products Inc.
Priority date: 1998-07-23
Filing date: 1998-07-23
Publication date: 2000-11-15

Abstract

Alatrofloxacinmesylat v podstatě prostý méně polárních nečistot, parenterální kompozice altrofloxacinmesylatu pro použití k léčení bakteriálních infekcí a způsobů čištění alatrofloxacinmesylatu.Alatrofloxacin mesylate is substantially free of less polar impurities, parenteral altrofloxacin mesylate composition for use in the treatment of bacterial infections and purification methods alatrofloxacinmesylat.

Description

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Alatrofloxacin parenteral compositions

Technical field

The present invention relates to naphthyridone alatrofloxacin mesylate substantially free of rr. antibiotic polar impurities / parenteral compositions of this alatrofloxa-cinmesylate and methods of purifying alatrofloxacin mesylate.

Background Art

Alatrofloxacinmcsylate is the naphthyridone antibiotic trovafloxaein related prodrug salt. Alatrofloxacinmcsylate (prepared by commercially available methods) is produced in very high purity. However, the alatrof loxacinrae-sylate prepared by these conventional methods contains minute less polar impurities which tend to precipitate from the parental compositions after standing indefinitely. These less polar impurities cannot be adequately removed by commercially viable methods such as recrystallization, standard flash silica gel flash chromatography, solvent / solvent extraction methods, or silica processing. certainty can be a reaction product, a well-known literary inventor has discovered that it has not been successfully removed by treating the raw material with a polystyrene resin produced by any of the other methods.

It has been known to date that the quinoline antibiotic cipro-floxacin also contains impurities causing this product to be unsuitable for parenteral formulations. European Patent Publication No. 237,226, published October 26, 1, S8 discloses the purification of ciprofloxaein by treatment with diatomaceous earth to obtain a product that is suitable for parenteral formulation. - 2 - ·· · · • • · · · · · · · · • · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · ·

As mentioned above, the processing of the alatrofloxacin mesylate by the silicone does not adequately remove less polar impurities.

Antibactrial activity and the synthesis of the alatroflo: cacin mentioned above are described in U.S. Patent Nos. 5,154,402 and 5,222,395 issued November 17, 1992, and July 20, 1993, the disclosure of which is incorporated by reference in its entirety. The foregoing patents are cited in conjunction with the present invention.

International Patent Publication WO 27/00253 published January 3, 1997 claims an alternative procedure for the preparation of alatrofloxacin. Thus, this publication is incorporated herein. by reference in its entirety.

International Patent Publication No. 97/03191, published March 1997, discloses a third process for the preparation and polymorphs of alatrofloxacin, which disclosure is incorporated herein by reference in its entirety. This patent publication is cited together with the present application.

International Patent Publication WO 95/19351, published July 20, 1995, U.S. Patent Application No. 50/021419 filed July 2, 1995 and International Patent Application No. PCT / EP9G / 04732 filed October 31, 1995, claim new intermediates for the preparation of alatrofloxacin and trovafloxacin, the disclosure of which is hereby incorporated by reference in its entirety. The foregoing patent publications and applications are cited together with the present application.

U.S. Patent No. 5,475,116, issued December 12, 1395 and U.S. Patent No. 5,255,791, issued October 25, 1993, claim various azabicyclohexane intermediates useful in the synthesis of alatrofloxacin and trovafloxacin, the disclosure of which is incorporated herein by reference in its entirety. - 3 - ♦ ♦ • • · · · · · · · · ·. The foregoing patents are assigned together with the present application. U.S. Patent Application 08 / 764,655, filed December 2, 19SS, relates to methods of treating Helicobacter pylori infections with alatrofloxacin and trovafloxacin, the disclosure of which is incorporated herein by reference in its entirety. The foregoing patent application is cited together with the present application.

International Patent Publication 26/33405, published December 12, 1925, claims a novel crystalline form of anhydrous 7- (1eC, 5o, 5C, GeC) -S-amino-3-azabicyclo [3.1.0] hex-yl) -6 -fluoro-1- (2,4-difluorophenyl) -1,4-dihydro-4-oxo-1,8-naphthyridin-3-carboxylic acid methanesulfonic acid salt. a method of using the compound of the invention in mammals, particularly humans, and the disclosure herein includes foi in the treatment of a bacterial infection-pharmaceutical composition whose reference in its entirety. The foregoing patent publication is cited together with the present application.

International Patent Application July 22, 1926 Application PCT / IB 9O / 0075é, claims the z-itterionic forms of trovafloxacin which are useful for administering a drug to a suspension, the disclosure of which is incorporated herein by reference in its entirety. is - 4 - · ♦ ♦ · ♦ • · • • • • • «« «« «« «« «« «« «« «« ♦♦ ·· ··

SUMMARY OF THE INVENTION

impurities involving the treatment of the impure reaction product containing said compound of formula (I) and less polar ones with a hydrophobic resin.

These less polar impurities are defined by their high pressure liquid chromatography (HPLC) retention times from sample injection to a Hewlett-Packard Model 1100 Series Kigh Pressure Liquid Chromatographer (R) eluting with a mobile phase solution of 40% acetonitrile, 0.05 # dihydrogenphosphonate Potassium (KI ^ PO, and 0.3¾ perchloric acid (v / v) at a flow rate of about 2 ml per minute to Puresil (r)

1 P 5 micron C silica HPLC column (4 µl0 mm) with 20 µl injection volume and Hewlett-Packard 1100 Series Diode Arrav Detector © with a detector set to record 270 wavelength. The HPLC column was used at ambient temperature (20-25 ° C). - 5 - ·· ·· ························ · ·· ·· ··

These less polar impurities defined by the above HPLC conditions have a retention time of about 2.1 minutes to about 30 minutes. A particularly undesirable less polar impurity has a retention time of 9 to 12 minutes under the above-mentioned HPLC conditions. The above-mentioned impurity with a retention time of 9 to 12 minutes has the formula II

also referred to herein as " oligomeric impurity ". Preferred less polar impurities after processing contain manna than 500 ppm, more preferably less than 60 ppm, relative to the weight of the crude product. More preferably, these impurities are less than 20 ppm. Preferably, the resin is CG-1S1®. a polystyrene-crosslinked hydrophobic resin such as Diaion HP-20 R or Ambsrchrom Presented

also refers to the compound of formula I · · · · · · · · · · · · · · · · · · · · · · · · · · · ·

- O

OH substantially free of less polar impurities / more preferably substantially free of the compound of Formula II. The present invention also relates to a parenteral composition comprising an antibacterially effective amount of a compound of formula I or a pharmaceutically acceptable salt thereof, substantially less less polar impurities, preferably substantially free of the compound of formula II, and a pharmaceutically acceptable carrier. Preferably, the pharmaceutically acceptable carrier is water. The present invention also relates to a patented composition as described above wherein the compound of Formula I is a lyophilizate. The present invention also relates to a pharmaceutical composition as described above. While the compound of formula I contains about 10 mg to about 700 mg, preferably about 275 mg to about 500 mg of the compound in a unit dose container, preferably about 300 mg. refers to a compound of formula

II

The present invention, comprising the formula II or the invention, also relates to a pharmaceutical complex-effective amount of a pharmaceutically acceptable salt thereof. The present invention relates to a method of rial infection comprising administering to a subject a bacterial infection, administering an antibacterial compound of Formula II. The present invention also relates to a compound of formula III

The present invention also relates to a pharmaceutical composition comprising an antibacterially effective amount of a compound of formula III or a pharmaceutically acceptable salt thereof. The present invention also relates to a method of treating a bacterial infection comprising administering to a subject infected with a bacterial infection, an antibacterial effective amount of a compound of Formula III.

DETAILED DESCRIPTION OF THE INVENTION atr1 atrofloxacin can be described in published patents or in three ways that patent publications have been published. Specifically, the alatrofloxacin may be prepared by binding of (L-Ala-L-Ala) -azabicyclohexane with activated quinoline as described in US Patent 5,154,402 issued on Nov. 17, issued July 23, Topad 1S92 and US Patent 132

The second method for the preparation of alatrofloxacin involves the reaction of a phthyridine publication of SOC-L-Ala-L-Ala with a protected azabicyclohexane nucleus as described in International Patent '70 37/30253, published January 3, 1337. Finally, a third method of alatrofloxacin production involves binding of trcvafloxacin mesylate with BOC- L-Ala-L-Ala as described in International Patent Publication UO J ' 7/03191, published March 6, 1397.

The publications of any of the above patents are incorporated herein by reference in their entirety. Q - The foregoing patents and publications are cited together with the present application.

Crude alatrofloxacin prepared by any of the above methods contains small amounts of less polar impurities. The specific type and amount of impurity depends on the process and conditions used. The crude alatrofloxacin drug product prepared according to each of the above three procedures has characteristic HPLC chromatograms when recorded according to the above conditions as shown in Figures 1-3.

Specifically, Fig. 1 describes a chromatogram of alatroflorcacin masylate prepared by the methods described in Brighty et al., U.S. Patent 5,164,402, issued November 17, 1992, and U.S. Patent 5,229,336, issued July 20, 1393. Approximate concentration of impurities of this original method when normalized to well known oligcmer concentrations are shown in Table 1 below.

FIG. 2 depicts an HPLC chromatogram of alatrofloxacin mesylate prepared by the method described in International Patent Publication 770 97/00258, published January 3, 1397. The approximate concentration of impurities produced by this second method when normalized to the known oligomer concentration is shown in Table 2 below.

FIG. 3 depicts the MPLC chromatogram of alatrofloxacinme-sylat prepared by the methods described in International Patent Publication 770 97/00191, published March 6, 1997. The approximate concentration of impurities produced by this third method when normalized to a known oligomer concentration is shown in Table 3 below. - ♦ * ·· f · • · · · ·

As stated above, the less polar impurities are defined by their retention time from sample injection in the Hewlett-Packard Model 1100 Series High Pressure Li-quid Chromatograph, eluting with a mobile phase solution of 40% acetonitrile, 0.0511 potassium dihydrogen phosphate (KH 2). PO *) and 0.3% perchloric acid (bbj / v) at a flow rate of about 2 ml per minute on Puresil® Smicron C silica column (4.5.150 mm) with a 20 microlitre injection volume and an Ilevette-Packard 1100 Series Diode Array Detector T T ) with a detector set to record at a wavelength of 270 nm.

Said less polar impurities defined by the above conditions have a retention time of about 2.2 minutes to about 30 minutes. One particularly undesirable impurity has a retention time of mxnc of 12 minutes (approximately 10.5. Above-mentioned impurity minutes) under the above-mentioned retention time of 9 to 12 minutes

has formula II

The impurity contains about ppm ppm of the reaction product formed by the third method described above. The absolute and applied conditions, less than about 100 ppm 50 ppm, most preferably the amount of impurity depends on the process. After cleaning, the TiO 2 impurity contains the reaction product, preferably less than 20 ppm. - 11 - - 11 -

Other impurities have retention times of about 23.0, 20.1, 13.5, 11.7, 11.0, 10.3, 9.3, 8.4, 7.5, 7.1, 6.2, 5.5, 5.0, 4.8, 3.9, 3.5, 3.1, 2.8, 2.5, and 2.1 minutes, as described above. Compounds corresponding to this retention regimen have not yet been characterized by their low concentration. these impurities can be removed by resin chromatography eluting with a polar solvent. In particular, the reaction product is dissolved in a small amount of water. The dissolved crude reaction product is then added to a column filled with polystyrene resin, such as hydrated Amberchron CG-IoK®, polystyrene resin (Tosohaas Chemicals 155 Kaystona Drive, Ilontgomery Mily, PA 18335), Amberlite (r) XAD-4 XAD-15 ( Rohm and Haas) or Diaion-IP-20 (r) (Mitsubishi Chemical Co), preferably hydrated Anberchrom CG-1S1 R polystyrene resin.

Polystyrene rubber styrene-based resin-shaped non-polar aromatic experts know that yories are hydrophobic spherical-type macroporous spheres that emboss surfaces for adsorption. Barbecue nonionic polymer resins are available in a variety of ball sizes, pore sizes and surface areas. Those skilled in the art will be aware of separation conditions, that resin parameters may be shirred to optimize results for particular applications.

The weight ratio of crude reaction product (kg) to dry resin weight (kg) is from 8 to about 15 kg of crude reaction product to about 1 kg of dry resin. Preferably, the weight to resin ratio is the crude reaction product ratio: about 10 to 12 kg of crude reaction product

The product per kg of dry resin.

The volume ratio of the eluent (liters) to the weight of the raw drug substance (kg) varies from 1 to about 70 liters of solvent per kg of drug substance, preferably 10 to 40 liters of water per kg drug substance at ambient temperature. Most preferably, about 25 liters of water per kg of drug substance at ambient temperature elutes the pure product. The eluent flow through the column is in the range of 1 to 10 layer volumes per hour (i.e., the amount of eluent required to fill and empty the resin layer at 1 hour). Preferably, the flow of eluent solution is 3 to 5 layer volumes per hour.

The product prepared by the methods described above is substantially free of less polar impurities and is suitable for use as a parenteral composition. Purified alatrofloxacin mesylate, where the starting materials are crude alatroflonacin drug products prepared according to the above three procedures, have HPLC chromatograms when recorded according to the above conditions as shown in Figures 4 to 5.

In particular, Fig. 4 shows a chromatogram of purified alatroflooccine mesylate after resin processing prepared by treating crude alatroflo: cacinmesylate prepared according to the methods described in Brighty et al., U.S. Patent 5,164,402 issued November 17, 1992 and U.S. Patent 5,229,3SS issued July 20, 1393 The approximate concentration of impurities after purification when assigned to a known amount of oligomer is presented in Table 1 below.

FIG. 5 discloses an I-IPLC chromatgram of a purified alatrof ionacin mesylate prepared by treating a crude alatrof loracin mesylate prepared according to the procedures described in International Patent Publication WO 97/00258, published January 3, 1997. 13 ·· · · · · · · · · · e · What is approximated to the concentration of impurities after purification when the known amount of oligomer is shown in Table 2 below.

FIG. 6 depicts an HPLC chromatogram of purified alatro-flonacin mesylate prepared by treating alatrofloxacin mesylate prepared according to the procedures described in International Patent Publication No. 0 97/03191, published March 5, 1997. The approximate impurity concentration after purification when normalized to a known amount of oligomer is shown in the following Table 3. 1

Table

Retention time (minutes) Pre-treatment concentration. (ppm) concentration after treatment (ppm) m / z 214. • ito 250 1 3.1 197 b 3.5 94 - 0 w / ✓ 35 -. ·: &Gt; / o 15. - 5.0 17 - 5.5 17 - 5.2 33 - / / 0 27 - O 0 ^ fV 30 - O u 10.9 10 20.1 211 - 23.0 23 - 14 - · '· ·· ............ · · · · · · · · · · · · ·

Table 2

Retention time Pre-treatment concentration. concentration after processing (minutes) (ppm) (pan) 0 i Z, A. o * > ouz 2.5 234 3.1 77 2 5.5 7 6.2 55 7.1 IQ 7.6 113 O Λ A o, ·. · *** 9/8 76 11.7 5 13.6 C 20 , 1 4S

Table 3 Retention time. Concentration before processing concentration after processing (minutes) (ppm) (ppm) 2.1 23 2.5 O 2.3 5 3.1 33 5 5.0 5 5.5 124 13 O / i. 22 gt > · O · # ť- O υ 10,5 700 11,0 97 20,1 22 · - 15 - ψ · · • ·

Parenteral compositions are more affected by trace contaminants than other pharmaceutical compositions. This is because parenteral compositions, whether diluted or reconstituted, are dosage forms whose quality standards are obvious to visual inspection. Visual inspection of parenteral compositions may record less polar impurities of up to 10 ppm. The presence of visible impurities affects the public's belief in safety, stability and product use. It is therefore important to remove as many less polar impurities as possible from parenteral compositions because less polar impurities are generally less water soluble and more likely to precipitate from the formulation. Further, most industrialized countries require that parenteral compositions suitable for treatment achieve a minimum standard for purity and absence of particulate contamination for drug approval.

The minimum standard in the United States is that the parenteral composition is substantially free of particles that may be detected by visual inspection, United States Pharmacopeia, R-24S (129d) - United States Pharmaco-psia (USP) does not create concretization for a minimum amount of particulate matter, that is, impurities that meet this standard. The USP provides some tests to quantify sub-visible foreign particles that would be ancillary to qualifying the amount of particulate matter. However, none of these tests is sufficient to meet the standard that the composition should be essentially free of particles that may be detected by visual inspection ". Although none of the assays are accepted as assays confirming the purity of the composition, turbidity analysis is one of the commonly accepted measurements to quantify the presence of particulate matter, see UJPassl et al., Analyst, 105,512-515 (1900). - 15 - · · · · · · · · · · · 9 9 9 9 9 · 9 · 9 9 9% * 999 99 99 999

According to standard turbidity measurements, the 0 - 0/45 non-cellular haze (IITU) value is accepted as a particulate-free solution.

The product of the pre-annealed invention, when the oyl is analyzed by a turbidometric device (Helligsurbiaimster, (877 Stewart Avenue / Garden City, USA; York 11530), routinely obtained low 0, UXU turbidity measurements. The crude starting material produced by the method described in International Patent Publication 110 97/08101 obtained a zakal measure of about 1.0 µm.

FIG. 7 shows the relationship of turbidity to the function of the concentration of impurities (ppm of oligomer in relation to the raw material). The process of the present invention includes all compositions having a base measurement of less than 0.5 KTU- changes in dosage will be made and the sub state is. which will be in any case

The thus purified product may be formulated for parenteral administration (e.g., intravenous, intramuscular, or subcutaneous) to mammals, including humans, for the treatment of bacterial infections by procedures well known to those skilled in the art or as described below. about 0.5 to about 1 g mg / kg / day, preferably about 4 to about 7 mg / kg / day, a negative of about 4.5 mg / kg / day. However, it is also a person who is responsible for dosing an appropriate dose for an individual subject.

The alatroflonacin mesylate of this invention can be administered widely by a variety of different dosage forms, generally the therapeutically active compounds of this invention are present in these dosage forms at concentration levels in the range of about 0.5 to about 70% (100S for lyophilic). - 17 - · · · · · · · · · · · · · · · · · · · ·

For parenteral administration (intramuscular, intraperitoneal / subcutaneous and intravenous use), a sterile pyrogen-free injectable solution of the active ingredient is customarily prepared. Solutions of the compound of formula I substantially free of the polar impurities, preferably compounds of formula II, can be prepared in a solvent such as sesar oil, peanut oil, water, or aqueous propylene glycol. the preferred solution is an unbuffered aqueous solution.

Aqueous solutions can be suitably adjusted and buffered, if necessary, and the liquid diluent is isotonic. These aqueous solutions are suitable for intravenous injection purposes. Oil solutions are suitable for intra-articular intramuscular and subcutaneous injection purposes. The preparation of all these solutions under sterile conditions is readily accomplished by standard pharmaceutical techniques well known to those skilled in the art. In the case of animals, the compounds may be administered intra-muscularly or subcutaneously at dosage levels of about 0.1 to 20 mg / kg / day, preferably about 0.2 to about 10 mg / kg / day given in a single dose or up to three divided doses.

Also, in vitro, the antibacterial agents of formula II, when hydrolyzed in the L-Ala-L-Ala-dipeptide moiety, have trial activities. This is hydrolyzed by salt

the compounds have the formula III - 13 - · · · · · · · · · · · · · h2n

OH

(HIM

The antibiotic activity of Formula III can be determined by methods well known to those of ordinary skill in the art, such as those described in S. Steers et al., Antibiotics and Chemotherapy, 9, 307 (19). The compound of formula III, when tested according to the method described above, gave a minimum inhibitory concentration (MIC) of 25 microg / ml to Streptococcus pyogenes and 12.5 to 25 microg / ml to numerous strains of Staphylococcus aureus. The compound of formula III may be formulated by a method using one or more compounds; / carriers. The maca can be prepared for oral, buccal or rectal administration.

For oral administration, a pharmaceutical composition of a compound of formula III may take the form of, for example, tablets or capsules prepared by conventional means with pharmaceutically acceptable excipients such as binders, (e.g., pregelatinized corn starch, polyvinylpyrrolidone or hydroxypropyl ethylcellulose), fillers (e.g., lactose, microcrystalline cellulose or phosphate) calcium), lubricants (e.g. magnesium stearate, talc or silica), disintegrants (e.g., potato starch or sodium starch glycolate), wetting agents (e.g., sodium lauryl sulfate) - - 19 - f · · · t · ... and

Liquid preparations for oral administration may take the form of, for example, solutions, syrups, or suspensions, or may be presented as a dry product for constitution with water or other suitable vehicle before use. Such liquid preparations can be prepared by conventional pharmaceutical compositions. acceptable acceptable additives such as suspending agents (for example, sorbitol syrup, methyl cellulose or hydrated edible fats), emulsifying agents (e.g., lecithin or acacia), non-aqueous vehicles (e.g., almond oil, oily esters or ethyl alcohol) and preservatives (e.g., methyl or propyl p-hydroxybenzoates or seric acid).

For buccal administration, the composition of the compound of formula IU may take the form of tablets or lozenges formulated in a conventional manner.

The compound of formula III may also be formulated in rectal compositions such as suppositories or retention enemas, for example, containing conventional rosehip bases such as i. cocoa butter or other glycerides.

The proposed dosage of a compound of formula III for oral, rectal or buccal administration to an average adult human for treating a bacterial infection is between about 0.1 to about 0.5 to a dosage unit of about 500 mg / kg / day of active ingredient to about 50 mg / kg / day and could be 4 times a day. served, for example, lx to - 20 -

Examples of carrying out the invention are not corrected in the compound. (d) and the NHA composition is the following examples illustrate the present invention. The melting points are e in parts per million referenced to the dautarial stabilization signal from the sample plate (deuteriodimathylsulfoxide unless otherwise specified).

Commercial reagents were used for further purification, referring to H, β-dimethylformamide. Chromatography refers to high pressure liquid chromatography of Hewlett-Packard model 1100 Series High Pressura Liquid Chromatograph (§) eluting with a mobile phase solution of 40µl acetonitrile, 0.01: dinyldrogenphosphorous potassium (KH 2 PO 4) and 0 3 µl perchloric acid (v / v) at a flow rate of about 2 ml per minute to Puresil (1 µm) micronized silica HPLC quartz (4.6.150 mm) with a 20 microliter injection volume and Ξ T1. - / lett-Packard 1100 Series Diode Array Dstector (r) with cict O '. : torque set to record at 270 nm. The column is used at ambient temperature. Room temperature or ambient temperature refers to 20-25 ° C. All non-reactive reactions were performed under nitrogen for comfort and maximum yields. Concentration under reduced pressure means that rotacap evaporator is used. Example 1

Batch resin processing of alatroflouacin mesylate

Alkaline acyl mesylate (50 g) containing approximately 700 gpm of oligomeric impurity in addition to other less polar impurities was dissolved in 0.05 µl aqueous methanesulfonic acid (714 µl) and then Mitsubishi Diacon phi-20 ($) hydrophobic resin (50 g) was added . - 21 -

After stirring the resin slurry for 24 hours in the dark, the slurry was filtered and the solution was analyzed by high pressure liquid chromatography (HPLC). (HPLC conditions were as described above.) The filtered solution contained 19 ppm of oligomeric impurity at 80% recovery of aiatrofloxacin mesylate.

Olimer had the following HMR (1e 5MR DMSO dg) S: 2.9 3 (d, 1H), 8.79 (s, III), 8.51 (s, 1H), 8.45 (d, 1H), 8 , 16 (d, 1H) 3.03 (d, 1H), 7.92 (d, 1H), 7.7 ° (t, 2H), m / z 1 (m, 2H), 7.33 ( m, 2H) 4.18 (g, 1H), 3.7 7 (q, 1H), 3.60 (m, Ul-1), 2.54 (s, 1H), 2.34 (3.111) 2.27 (3.311), 1.92 (bs, 211), 1.72 (b3.2Ii), 1.28 (d, 3i1), 1.17 (d, 3j Example 2 Resin Column Processing aiatrof loxacin mesylate

Aiatrofloxacin mesylate was dissolved in 10-40 L water per kg drug substance at ambient temperature. The solution was filtered and then passed through (3 to 5 layer volumes per hour) column containing hydrated Amberchrom CG-polystyrene resin (about 10 to 12 kg of crude reaction product per kg of dry resin). The column was washed with water (3 L per kg of crude reaction product) and then with a reagent and the washings were combined. The processed alla-trofloxacin mesylate solution was analyzed by HPLC. The polystyrene resin column treatment reduced the level of impurity of formula II from 500 ppm to less than 30 ppm relative to the drug, in addition, the drug was obtained in 95% resin yield. Example 3

Insulation ·· · 9 · 9 · 9 · 9 9 9 9 9 9 9 · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · ·

· © • •

A solution containing the resin-treated alatrofloxacinmesylate (10 g) (prepared in Examples 1 or 2), deionized water (154 ml) and ethanol (31 ml) was neutralized to a pH of about δ / 5 to 7.5 by dropwise addition of 10% aqueous lye. The suspension was granulated for 30 minutes at 24 ° C and then filtered. After solids were washed with ethanol (30 mL), the zwitterion was dried under vacuum at 40 ° C for about 1 hour. Alatrofloxacin (800 g) was obtained in 95% yield as white trapezoidal crystals. Example 4

Creating a1a tro f1oxad i nme with y1a here

Alatrofloxacin zv / itterion (25 g wet cake) as beech wet cake or dry product was combined with 10% aqueous ethanol (242 mL) and then methanesulfonic acid (1.9 mL) was added to the slurry to adjust pH to about 3.5 to 4.5. The reaction mixture was then heated to reflux, filtered and then concentrated at atmospheric pressure to about 1/2 of its original volume. The solution was then cooled to 0-5 ° C and the resulting solid was granulated for 1 hour. After filtering the solids and washing with ethanol (39 mL), the alatrofloxacinmaslate was dried for 2 hours at 40 ° C. Alatrofloxacinmasylate (17.7 g) suitable for parenteral formulation was isolated in 90% yield. 5 i'l. lc *. ^

ParGrial compositions in vials

Alatrofloxacin mesylate (314.44 mg) prepared according to Example 4) was dissolved in a minimum amount of water for injection, United States Pharmacopeia (WFI). - 23 - ·· M ** • · »« • • • • • • • • • • • • • • • • • • * t fl • · • • • • • •. · · · · · · · · · · · · · · · · · · · · · ·

The pH of the resulting solution was adjusted to pH 4 with a 4% w / w hydrochloric acid solution, National Formulary (sJF) in water for injection and / or lazy / w / w sodium hydroxide solution, Ilatinal Formulary in the injection compartment and diluted to 7.85 mg / ml. The resulting solution (4.0.12 g) was filtered and filled into a 40 mL vial and stoppered. The stoppered vial was autoclaved at 115 ° C for 15 minutes. Example 6

Parenteral composition in ampoules

Alatrofloxacin mesylate (314.4 mg) prepared according to Example 4 was dissolved in a minimal amount of water for injection, United States Pharmacopeia (VFI). The pH of the resulting solution was adjusted to 4 with a lazy nm / wt solution of hydrochloric acid, National Formulary (UF), in water for injection, or lazy w / w sodium hydroxide solution, National Formulary, in water for injection and diluted to a concentration of 7, 85 rng / ml. The resulting solution (40.12 g) was filtered and filled with a 40 mL ampoule and sealed. The sealed vial was sterilized by autoclaving at 115 ° C for 15 minutes. Example 7

Lyophil

Alatrofloxacin mesylate (314/4 mg) prepared according to Example 4) and lactose (500 mg, National Formulary) were dissolved in sufficient water for injection, United States Pharmacopeia to give a final weight of 10,000 mg. - 24 - Μ ·· · * · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · • The IT bottle was lyophilized for 24 hours under a vacuum of 0.1 atmosphere and then stoppered and sealed. The sealed vial was sterilized by autoclaving at 115 ° C for 15 minutes. Example 8

Isotonic parenteral composition in vials

Daxtrose (500 mg, United States Pharmacopeia) and alatrofloxacimesylate (314.46 mg, prepared according to Example 4) were dissolved in a minimum amount of water for injection (United States Pharmacopeia) and pM was adjusted to 4 µl / wt hydrochloric acid solution (National Porularya) in water for injection and / or a w / w solution of sodium hydroxide in water for injection (United States Pharma mg / ml. The resulting solution is 100 ml vial and sterilized by autopsy) and diluted to a concentration of 3.14 (101.7 g) was filtered and filled into a stopper. The stoppered vial was a key at 115 ° C for 15 minutes. Example

Isotonic parenteral composition in ampoules

Dextrose (500 mg, United States Ph trofloxacin mesylate (314.4 mg, prepared 4), was dissolved in the minimum amount of armacopaia) according to the water supply for the United States Pharmacopeia and the pH was adjusted to 4 by weight. / wt. of a solution of hydrochloric acid (phthalate formulations) in water for injection and / or with any sodium hydroxide solution in water for injection (United States Pharmacopeia) and diluted to 3.14 mg / ml. The resulting solution (101.7 g) was filtered and filled into a 100 ml ampoule and sealed. - 25 - - 25 - ♦ · · · 0 · · · · · · · · · · · · · · · · · · ff • · · · · · · · · · · · ·

The sealed vial was autoclaved at 115 ° C for 15 minutes. Example 10

L-Alanyl-N - [3- [5 - [[[3- [S-carboxy-3- (2,4-difluorophenyl) -3-fluoro-5,8-dihydro-5-oxo- 1,3-naphthyridin-2-yl] -3-azabicyclo [3.1.0] hex-5-yl] arauro] carbonyl] -3- (2,4-difluoro-phenyl) -3-fluoro- 5,3-dihyro-5-oxo-1,3-naphthyridin-2-yl [-3-azabicyclo [3.1.0] hex-5-yl] -L-alaninamide 2 5Qml single neck round bottom flask equipped with re -flux cooler and magnetic stir bar. The flask was rinsed and then maintained under a dry nitrogen atmosphere, othvlaster (1g4, 0C, ogC) - 7 - (o - ciuuino - 3 - azabicyiclo - [3.1.0] hex-3-yl) -1 " -difluorophenyl) -δ-fluoro-1,4-dihydro-4-oxo-1,8-naphthyridine-3-carboxylic acid (2.50 g, 5.62mmol) (prepared according to the method described in International Patent Publication | (1 S, 5 S, 6 S) - H - [(1,1-Dimethyl-ethoxy) -carbonyl] -L-alanyl-N-N '- [3- (2,4-difluorophenyl) -S- carboxy-3-fluoro-5,3-dihydro-S-oxo-1,3-naphthyridin-2-yl] -3-azabicyclo- [3.1.0] hex-3-yl] -L-alanine amide (3.37) g, 5.11 mmol) (prepared as described in International Patent Publication '7097 / 08191) and 100 ml of methylene chloride in this flask to give a white ivy. Then, 4-dimethylaminopyridine (0.37 g, 3/07 mmol) and bromotrispyrrolidine phosphonium hexafluorophosphate (2.35 g, 6.14 mmol) were added to the reaction mixture, and 1.3 mL of diisopropyl-thiianamine (10 mL) was added slowly. 2 mmol) over 30 minutes. Overnight • The reaction mixture was stirred at 25-30 and then thin layer chromatography (silica gel silica gel, 5.10 cm silica gel, using ammonium hydroxide / methanol / methylene chloride (1:10:30)) showed that the reaction it was complete. ·· 9 • 9 9 · 9 · 9 4 9 9 4 9 9 99 - 26

The thick slurry was filtered, the filter cake was washed with methylene chloride (2.20 mL) and then dried under vacuum at 25 to 20 ° C to give 4.53 g (33% yield) of an oxo-protected intermediate, a compound of formula II. (CDCl 3, 1 H, 8.58 (s, III), 8.3 5 (s, III), 8.02 (d, J = 12.7 Hz, III), 7.95 (d, 1 H); J = 12.5: 1H, III), 7.33 (m, 2H), 7.05 (m, 1H), 6.70 (m, 1H), 5.24 (m, 1H), 4.39 (q, J = 7.25 Hz, III), 4.33 (q, J = 7.2 5 Hz, 2 K), 4.03 (q, J = 1.7 µz, 11 I), 3.8 7 (m, H), 3.55 (m, 4H), 2.55 (s, 1H), 2.40 (s, 1H), 1.87 (m, 411), 1.41 (s, 9H), and 1.35 (m, 3H). FA2MS: m / z 1086 (MH +).

Hethane sulfonic acid (1.4 mL) and water (3.1 mL) were added to the bipolar intermediate (4.53 g, 4.23 mmol) suspended in 135 mL of 1-methyl-2-pyrrclidinone. The solution was formed after heating the reaction mixture for 15 minutes at 80-85 ° C and the reaction was substantially complete after heating for a further 48-58 hours at 80-85 ° C.

The solution was cooled to room temperature and 270 mL of methylene chloride was added slowly. The resulting slurry was stirred for 2 days at room temperature and then filtered. The solid was dried under reduced pressure to give 4.0 g of crude product. The granular solid was crushed with a pestle in a dowel and fired suspended in 150 ml of methylene chloride. After stirring the slurry for 24 hours at room temperature, the solid was filtered and then air dried to give compound II as a mesylate salt (3.20 g, 75% yield). 1H (IR) (DMSO-d-): 00.93 (d, 1H), 3.7 (s, 1H), 8.51 (s, 1H), 3.45 (d, 1H), 8 1S (d, 1H), 3.03 (d, J = 12.5Hz, 1H), 7.92 (d, J = 12.9Hz, III), 7.75 (m, 2H), 7, 51 (m, 2H), 7.33 (m, 2H), 4.13 (q, J = S, S5Hz, 1H), 3.77 (q, J = 7.05Hz, 111), - 27 - • 3.00 (01.811), 2.54 (s, 1H), 2.34 (s, 1H), 2.27 (s, 3H), 1.92 (s, 2H), 1.72 ( s, 211), 1, 28 (d, J = 0, 35 Hz, 3I) and 1.17 (cl, J = 7.06 Hz, 3H). PABMS: m / z 957 (1HCl 3 SO 3 H). Example 11

Ilethanesulfonate (1, 5, Gtx) -7- (5- (S-amino-3-azabicyclo [3.1.0] 113: 3-yl) -1- (2 4-difluorophenyl) -5-fluoro-4-one-1,4-dihydro-1,8-naphthyridine-3-carbonamide] - (1 << 5 >, 5λ) -3-azabi- cyclo [3,1,0] hen-3-yl) -1- (2,4-difluorophenyl) -5-fluoro-4-oxo 1,4-dihydro-1,8-naphthyridine-3-carbonyl acid

Into a 20 ml flask with a 1. 1: .th in the bottom of the flask, which was fitted with a fluctuating target, the ball was poured into the ball, and the ball was driven. (lck, 5c < 5, < 1 >) -1- (2,4-difluorophenyl) 7- [c- [[(1, 1-diethyl) r < / RTI > - [3, 1, 3] cyano-3-yl) -5-fluoro-1,4-dihydro-4-oxo-1-yl-phthyridine-carboxylic acid (2 2 2 mg, 0.45 mmol) (prepared according to U.S. Pat. No. 5,154,402), triethylamine (1.14 g > 11.3 mmol), 2.0 ml of 50% w / w; This is a cycle of 1-prop and Ώ.Σ O-oxides (1, 43 O 4, 4.50 nu: ol) in thylacetate and 10 ml of tetrahydrofuran (vli). F). The resultant suspension was maintained under an inert atmosphere of a separate flask equipped with a magnetic stir bar and an ethyl ester suspension prepared by a pressure equalizing addition funnel (1 < *, 5 <<<<<<<<<<< 7- (5-amino-3-azabicyclo [3.1.0] hept-3-yl) -1- (2,4-difluorophenyl) -5-fluoro-1,4 -dihydro-10-ol, O-naphthyridine-3-carbonylic acid (200 mg, 0.43 nmol) (prepared as described in International Patent Publication No. S7 / Q02G3), in 5 ml Γ1Γ. The yellow triethylamine-containing slurry was poured into an addition funnel and then slowly added over 20 minutes to the flask containing the ethyl ester (5i, 5C, 7C) -7- (6-amino-3-azabicyclo [3.1.0]). hex-3-yl) -1- (2,4-difluorophenyl) -fluoro-1,4-dihydro-4-oxo-1,5-naphthyridine-3-carboxylic acid. The mixture was stirred for 5 hours at room temperature when thin layer chromatography [Kieselgel GQ silica plate (5.10 cm) using matlianol / mathylene chloride (1: 9)] eluant showed the reaction to be complete. The yellow suspension was poured into 70 mL of water and the pH adjusted to 3.0 with hydrochloric acid. The aqueous solution was extracted with 70 mL of methylene chloride and then the methylene chloride solution was washed with water (70 mL), the methylene chloride was removed under reduced pressure to give a crude solid (449 mg) which was triturated with 5 mL of ethanol for G hours. The solid was washed with 2 mL of ethanol and then dried in vacuo for 18 hours. p i- 1 ~ 'Ολο, c j_ms 0 get. 1 o 3 04 mg (72% yield) of two-fold protective yarn in the form of a white crystalline solid. M.p .: 2: -251 ° C. 1íí (CDC 1) · X3 10.0 (d, 2Π), 3.6 0 (s, 1H), 3.35 (Ξ, 111), 8.05 (d, lil). 7.3 (c, 1H) 7.3 3 (m, 211), 7.0 5 (m, η K), 4.70 (r, 1H), 4.35 (q, 2H), 5.82 (m, 311) 5.0 o (m, from), z, o 3 (s, 1H), 2.23 (ε, 111), 1.85 (m, 2K), 1.77 (m, m) 211) 1.5 3 (s, 511) and 1, 3 8 (t, 311) r * S: ra / z 843 (li JL ') · i latii ar.su Ifon's acid in (0, 07 ml) and water (1, 1 µ ml) were added to the two-dose buffer between oroduk here (240 mg, 0/3 mmol) with a suspension and not in 7 ml methy _ methyl L 1-2 py rr clidi nonu. The reaction mixture was heated for 50 to 58 hours at 80 to 85 ° C. The solution was then cooled to room temperature and 14 mL of methylene chloride was slowly added. The resulting slurry was stirred for 2 days at room temperature and pal; filtered. Solid and material were dried under reduced pressure to give 110 mg of crude product. The crude solid was triturated with 3 mL of methylene chloride / methartol (1: 1) for 4 hours. O,

The solid was dried under vacuum at 45 to give the compound of the formula III as a precipitate salt (105 mg, 45% crude yield). Melting point above 230 ° C. II NMfl, recorded in trifluoroacetic acid, showed the absence of 30C and ethyl ester protecting groups and was consistent with the structure of compound III. FABMS: m / z 815 (ΐΓ ° Γΐ3303Η)

Claims

1. A method of purifying a compound of formula I

OH characterized by treating an impure reaction product containing said compound of formula I and less polar impurities with a hydrophobic resin.

2. The process of claim 1 wherein the less polar impurities have a retention time of about 2.1 minutes to about 30 minutes.

3. The process of claim 1 wherein the less polar impurities have a retention time of about 9 to about 12 minutes.

4. The process of claim 1 wherein one of said less polar impurities is of Formula II

- 31 -

5. The method of claim 1 wherein said non-post-processing purities comprise less than 50 porn in relation to 3: total weight of purified product.

5. The process of claim 1 wherein the post-treatment impurities include less than about 20 ppm relative to the total weight of the purified product. /. 2. A method according to claim 1, wherein; the resin is crosslinked with polystyrene resin. The process of claim 1 wherein the hydrophobic resin is Diaion P-20 R or Amberchrom CG-R. A method according to claim 3, characterized in is a Diaion HP-20 p or an Arab 10 compound of Formula I

11. A compound of the formula according to claim 10 wherein said I is substantially free of a compound of formula II.

12. Parentor antibacterially less polar or pharmacologically susceptible, characterized in that it contains an effective amount of a compound of the formula: acceptable salt, substantially purifying, and a pharmaceutically acceptable carrier.

12. A parenteral composition comprising an antibacterial effective amount of a compound of formula I or a pharmaceutically acceptable salt thereof, substantially free of a compound of formula II, and a pharmaceutically acceptable carrier. 15. The parenteral composition of claim 10 wherein the compound of Formula I is a lyophilate. 15. The parenteral composition of claim 11, wherein the compound of Formula I comprises about 10 mg to about 700 of the compound in a unit dose container. 33

A parenteral composition according to claim 11, wherein the compound of formula (I) contains from about 275 mg to about 500 mg of the compound in a unit dose container.

13. A parenteral composition according to claim 10 wherein the compound of Formula I comprises about 10 mg to about 700 mg of the compound in a unit dose container.

13. A parenteral composition according to claim 10, wherein the compound of formula (I) comprises about 275 ng to about 500 m-c of the compound in a unit dose container.

20. A compound of formula II

i · ar me omo ozic2 v v z n a c e na im. that it comprises an antibacterially effective morning 20 or a pharmaceutically acceptable compound thereof according to an acceptable salt. claim

22. A method of treating a bacterium that is administered to a subject affects a bacterially effective amount of other infection characterized in that the bacterial infection of the compound according to claim 20-34 is administered to the subject.

A compound of formula III

25. A method of treating a bacterial infection, characterized in that an anti-bacterially effective amount of a compound of claim 23 is administered to a subject afflicted with a bacterial infection.

pup: