FORM 2
THE PATENT ACT 1970
(39 of 1970)
&
The Patents Rules, 2003
COMPLETE SPECIFICATION
(See section 10 and rule 13)
1. TITLE OF THE INVENTION:
POLYMORPHS OF S-(-)-9-FLUORO-6,7-DIHYDRO-8-(4-L-ALANINYLOXYPIPERIDIN-l-YL)-5-METHYL-l-OXO-lH,5H-BENZO[I,J] QUINOLIZINE-2-CARBOXYLIC ACID METHANESULFONIC ACID SALT
2. APPLICANT (S)
(a) NAME: WOCKHARDT LTD.
(b) NATIONALITY: INDIAN
(c) ADDRESS: Wockhardt Limited, D4-MIDC Area, Chikalthana,
Aurangabad - 431 210 (M.S.) INDIA.
3. PREAMBLE TO THE DESCRIPTION
The invention relates to crystalline and amorphous polymorphs of S-(-)-9-fluoro-6,7-dihydro-8-(4-L-alaninyloxypiperidin-1 -yl)-5-methyl-1 -oxo-1 H,5H-benzo[i j] quinolizine-2-carboxylic acid methanesulfonic acid salt and its pharmaceutical compositions. The polymorphs and compositions of the invention are useful to treat bacterial Gram-positive, Gram-negative and anaerobic infections, especially infections caused by resistant Gram-positive organism and Gram-negative organism, mycobacterial infections and emerging nosocomial pathogen infections. The following specification particularly describes the invention and the manner in which it is to be performed.


Field of the Invention
The invention relates to crystalline and amorphous polymorphs of S-(-)-9-fluoro-6,7-dihydro-8-(4-L-alaninyloxypiperidin-1 -yl)-5-methyl-1 -oxo-1 H,5H-benzo[ij] quinolizine-2-carboxylic acid methanesulfonic acid salt and its pharmaceutical compositions. The polymorphs and compositions of the invention are useful to treat bacterial Gram-positive, Gram-negative and anaerobic infections, especially infections caused by resistant Gram-positive organism and Gram-negative organism, mycobacterial infections and emerging nosocomial pathogen infections.
Background of the Invention
The chiral fluoroquinolone racemic (±)-9-fluoro-6,7-dihydro-8-(4-hydroxypiperidin-l-yl)-5-methyl-l-oxo-lH,5H-benzo[ij]quinolizine-2-carboxylic acid which is known as nadifloxacin is described in Japanese patents JP 63,192,753A and JP 05,339,238A. S-(-)-nadifloxacin is disclosed in Chern. Pharm, Bull 44 (1996).
U.S. Patent No. 7,247,642, which is herein incorporated by reference, discloses the S-(-)-9-fluoro-6,7-dihydro-8-(4-hydroxypiperidin-1 -yl)-5-methyl-1 -oxo-1 H,5H-benzo[i j] quinolizine-2-carboxylic acid L-arginine salt and hydrates thereof.
U.S. Patent No. 6,514,986, which is herein incorporated by reference, discloses the substantially amorphous and substantially crystalline forms of S-(-)-9-fluoro-6,7-dihydro-8-(4-hydroxypiperidin-l-yl)-5-methyl-l-oxo-lH,5H-benzo[ij] quinolizine-2-carboxylic acid L-arginine salt.
U.S. Patent No. 6,664,267, is herein incorporated by reference, discloses the crystalline monohydrate form of S-(-)-9-fluoro-6,7-dihydro-8-(4-hydroxypiperidin-l-yl)-5-methyl-l-oxo-lH,5H-benzo[ij]quinolizine-2-carboxylic acid L-arginine salt.


U.S. Patent No. 6,750,224, is herein incorporated by reference, discloses the prodrugs of S-(-)-9-fluoro-6,7-dihydro-8-(4-hydroxypiperidin-1 -yl)-5-methyl-1 -oxo-1 H,5H-benzo[i j] quinolizine-2-carboxylic acid. The patent '224 discloses two types of prodrugs - the prodrugs at the 2-carboxylic acid and at the 4-hydroxy of the piperidine. The prodrugs disclosed at 4-hydroxy piperidine include amino acid prodrugs. Amongst the amino acid prodrugs, L-alanine and L-valine prodrugs were disclosed.
U.S. Patent No. 7,164,023, is herein incorporated by reference, discloses the most stable salt form, S-(-)-9-fluoro-6,7-dihydro-8-(4-hydroxypiperidin-1 -yl)-5-methyl-1 -oxo-lH,5H-benzo[ij]quinolizine-2-carboxylic acid L-arginine salt tetrahydrate.
U.S. Patent publication No. 2007219227, is herein incorporated by reference, discloses the methane sulphonic acid salt of prodrugs of S-(-)-9-fluoro-6,7-dihydro-8-(4-hydroxypiperidin-1 -yl)-5-methyl-1 -oxo-1 H,5H-benzo[i j]quinolizine-2-carboxylic acid. The prodrugs disclosed include amino acid prodrugs.
The invention provides crystalline and amorphous polymorphs of S-(-)-9-fluoro-6,7-dihydro-8-(4-L-alaninyloxypiperidin-1 -yl)-5-methyl-1 -oxo-1 H,5H-benzo[ij] quinolizine-2-carboxylic acid methanesulfonic acid salt with improved characteristics such as stability.
Summary of Invention
The invention is directed to crystalline and amorphous polymorphs of S-(-)-9-fluoro-6,7-dihydro-8-(4-L-alaninyloxypiperidin-1 -yl)-5-methyl-1 -oxo-1 H,5H-benzo[ij] quinolizine-2-carboxylic acid methanesulfonic acid salt, of Formula I,



NH2 .CH3SO3H

o o

Formula I
wherein R is CH3.
The invention also relates to a method of making crystalline and amorphous polymorphs of S-(-)-9-fluoro-6,7-dihydro-8-(4-L-alaninyloxypiperidin-1 -yl)-5-methyl-1 -oxo-1 H,5H-benzo[ij] quinolizine-2-carboxylic acid methanesulfonic acid salt.
The invention further relates to pharmaceutical compositions containing crystalline and amorphous polymorphs of S-(-)-9-fluoro-6,7-dihydro-8-(4-L-alaninyloxypiperidin-l-yl)-5-methyl-l-oxo-lH,5H-benzo[ij] quinolizine-2-carboxylic acid methanesulfonic acid salt and to the method for treating or preventing bacterial infections using crystalline and amorphous polymorphs of S-(-)-9-fluoro-6,7-dihydro-8-(4-L-alaninyloxypiperidin-l-yl)-5-methyl-l-oxo-lH,5H-benzo[ij] quinolizine-2-carboxylic acid methanesulfonic acid salt and compositions of the invention.
Brief description of the figures
The invention is described by the following non-limiting examples which refer to the accompanying FIGS. 1 to 6, short particulars of which are given below.
FIG. 1 shows a representative powder X-ray diffractogram of crystalline polymorph of S-(-)-9-fluoro-6,7-dihydro-8-(4-L-alaninyloxypiperidin-1 -yl)-5-methyl-1 -oxo-1 H,5H-benzo[ij]quinolizine-2-carboxylic acid methanesulfonic acid salt


FIG. 2 shows a representative powder X-ray diffractogram pattern of amorphous
polymorph of S-(-)-9-fluoro-6,7-dihydro-8-(4-L-alaninyloxypiperidin-1 -yl)-5-methyl-1 -
oxo-lH,5H-benzo[ij]quinolizine-2-carboxylic acid methanesulfonic acid salt
FIG. 3 shows a representative Differential Scanning Calorimetric analysis of crystalline
polymorph of S-(-)-9-fluoro-6,7-dihydro-8-(4-L-alaninyloxypiperidin-1 -yl)-5-methyl-1 -
oxo-lH,5H-benzo[i j]quinolizine-2-carboxylic acid methanesulfonic acid salt
FIG. 4 shows a representative Differential Scanning Calorimetric analysis of amorphous
polymorph of S-(-)-9-fluoro-6,7-dihydro-8-(4-L-alaninyloxypiperidin-1 -yl)-5-methyl-1 -
oxo-lH,5H-benzo[ij]quinolizine-2-carboxylic acid methanesulfonic acid salt
FIG. 5 shows a representative Infra-red spectrum of crystalline polymorph of S-(-)-9-
fluoro-6,7-dihydro-8-(4-L-alaninyloxypiperidin-1 -yl)-5-methyl-1 -oxo-1 H,5H-
benzo[ij]quinolizine-2-carboxylic acid methanesulfonic acid salt
FIG. 6 shows a representative Infra-red spectrum of amorphous polymorph of S-(-)-9-
fluoro-6,7-dihydro-8-(4-L-alaninyloxypiperidin-1 -yl)-5-methyl-1 -oxo-1 H,5H-benzo[i j ]
quinolizine-2-carboxylic acid methanesulfonic acid salt
Detailed Description of the invention
In one general aspect, the invention provides crystalline and amorphous polymorphs of S-(-)-9-fluoro-6,7-dihydro-8-(4-L-alaninyloxypiperidin-l -yl)-5-methyl-1 -oxo-1 H,5H-benzo[ij] quinolizine-2-carboxylic acid methanesulfonic acid salt, of Formula I,
P 0


wherein R is CH3.

NH2 .CH3SO3H

Formula I


The crystalline polymorph of S-(-)-9-fluoro-6,7-dihydro-8-(4-L-alaninyloxypiperidin-l-
yl)-5-methyl-l-oxo-lH,5H-benzo[ij] quinolizine-2-carboxylic acid methanesulfonic acid
salt is characterized by characteristic X-ray diffraction pattern, Infra-red spectrum and
Differential Scanning Calorimetric analysis as follows-
X-ray diffraction pattern (26): 9.82 ± 0.2°, 11.76 ± 0.2°, 12.66 ± 0.2°, 13.38 ± 0.2°, 15.56
± 0.2°, 16.34 ± 0.2°, 17.58 ± 0.2°, 18.26 ± 0.2°, 19.74 ± 0.2°, 20.86 ± 0.2°, 21.70 ± 0.2°,
22.64 ± 0.2°, 23.58 ± 0.2°, 28.38 ± 0.2°, 33.18 ± 0.2°.
Infra-red spectrum (cm1): 3429, 2957, 1739, 1619, 1523, 1442, 1390, 1322, 1273, 1228,
1149, 1071, 1039,883,810.
Differential scanning calorimetric endotherm at 278.50 °C (onset at 275.92 °C).
An amorphous polymorph of S-(-)-9-fluoro-6,7-dihydro-8-(4-L-alaninyloxypiperidin-l-
yl)-5-methyl-l-oxo-lH,5H-benzo[ij] quinolizine-2-carboxylic acid methanesulfonic acid
salt is characterized by characteristic X-ray diffraction pattern, Infra-red spectrum and
Differential Scanning Calorimetric analysis as follows-
X-ray diffraction pattern (29): 19.86 ± 0.2°, 20.02 + 0.2°, 20.12 ± 0.2°, 20.28 ± 0.2°,
20.36 ± 0.2°, 20.74 ± 0.2°, 21.14 ± 0.2°, 25.36 ± 0.2°, 25.76 ± 0.2°, 25.94 ± 0.2°, 26.14 ±
0.2°, 26.28 ± 0.2°.
Infra-red spectrum (cm"1): 3446, 2959, 1738, 1621, 1528, 1451, 1393, 1327, 1205, 1047,
882,810.
Differential scanning calorimetric endotherm at 44.50 °C (onset at 42.50 °C), endotherm
at 160.50 °C (onset at 154.33 °C), exotherm at 202.66 °C (onset at 190.53 °C) and
endotherm at 272.00 °C (onset at 267.97 °C).
In another aspect the invention provides a method of preparation of crystalline polymorph of S-(-)-9-fluoro-6,7-dihydro-8-(4-L-alaninyloxypiperidin-1 -yl)-5-methyl-1 -oxo-1 H,5H-benzo[ij] quinolizine-2-carboxylic acid methanesulfonic acid salt, which includes the steps of;
a) reacting (2'S,5S)-9-fluoro-6,7-dihydro-8-{4-N-tert-butoxycarbonyl-L-
alaninyloxy-piperidin-1 -yl} -5-methyl-1 -oxo-1 H,5H-benzo[i j]quinolizine-2-carboxylic acid with methanesulfonic acid in ketone solvent to get crude S-(-)-9-


fluoro-6,7-dihydro-8-(4-L-alaninyloxypiperidin-1 -yl)-5-methyl-1 -oxo-1 H,5H-benzo[ij] quinolizine-2-carboxylic acid methanesulfonic acid salt,
b) contacting the crude S-(-)-9-fluoro-6,7-dihydro-8-(4-L-alaninyloxypiperidin-l-yl)-5-methyl-l-oxo-lH,5H-benzo[ij] quinolizine-2-carboxylic acid methanesulfonic acid salt with water,
c) treating the mass with ketone solvent and
d) isolating crystalline polymorph of S-(-)-9-fluoro-6,7-dihydro-8-(4-L-alaninyloxypiperidin-1 -yl)-5 -methyl-1 -oxo-1 H,5H-benzo[i j] quinolizine-2-carboxylic acid methanesulfonic acid salt
Crude S-(-)-9-fluoro-6,7-dihydro-8-(4-L-alaninyloxypiperidin-1 -yl)-5-methyl-1 -oxo-
lH,5H-benzo[ij] quinolizine-2-carboxylic acid methanesulfonic acid salt is prepared by the method known in the art such as U.S. Patent publication No. 2007219227. This crude material is dissolved under stirring in a water to obtain slightly hazy solution. The hazy solution is washed with acetate solvent which includes ethyl acetate and butyl acetate. The clear aqueous layer was concentrated under vacuum to provide a solid mass. The solid mass is added with equal volume of water under stirring, followed by ketone solvent (volume 12 times of the weight of solid mass). The suspension is stirred for 15 to 18 hours at 30-35° C and crystalline polymorph is isolated from the reaction mass.
Ketone solvent includes one or more from the group of acetone, Methyl isobutyl ketone, Di iso butyl ketone, Methyl iso propyl ketone and Methyl ethyl ketone.
In yet another aspect the invention provides a method of preparation of amorphous polymorph of S-(-)-9-fluoro-6,7-dihydro-8-(4-L-alaninyloxypiperidin-1 -yl)-5-methyl-1 -oxo-lH,5H-benzo[ij]quinolizine-2-carboxylic acid methanesulfonic acid salt. The process includes dissolving the crystalline polymorph of S-(-)-9-fluoro-6,7-dihydro-8-(4-L-alaninyloxypiperidin-1 -yl)-5-methyl-1 -oxo-1 H,5H-benzo[i j] quinolizine-2-carboxylic acid methanesulfonic acid salt with water followed by lyophilizing at freeze drying temperature. The isolated product is dried to get amorphous polymorph.


In yet another aspect the invention provides liquid and solid pharmaceutical formulations which comprise crystalline or amorphous polymorphs of S-(-)-9-fluoro-6,7-dihydro-8-(4-L-alaninyloxypiperidin-1 -yl)-5-methyl-1 -oxo-1 H,5H-benzo[i j]quinolizine-2-carboxylic acid methanesulfonic acid salt of the invention. The formulation includes injectable solutions, suspensions, emulsions, tablets, coated tablets, coated tablet cores, capsules, solutions, troches, dispersions, pellets, granules, suppositories, hard or soft gelatin capsules, and the like.
The pharmaceutical compositions are prepared according to conventional procedures used by persons skilled in the art to make stable and effective compositions. Such methods include by mixing, stirring, suspending, dispersing, emulsifying, dissolving and the like, the active compounds with or in the pharmaceutical auxiliaries such as a carrier, diluent, solvent or excipient and processing the components to pharmaceutically suitable forms for parenteral, oral, intranasal, buccal or rectal administration and the like. In the solid, liquid, parenteral dosage forms, an effective amount of the active compound or the active ingredient is any amount, which produces the desired results.
The dosage forms can be prepared by any conventional techniques recognized in the art, but would preferably be formulated by mixing the polymorphs of the invention with other ingredients. The other ingredients utilized to formulate solid oral dosage forms would include conventional inert ingredients such as microcrystalline cellulose, methyl cellulose and the like, suitable sweetening and/or flavouring agents, and preservatives thereof if required.
Such solid oral dosage forms or dry formulations suitable for the preparation of suspensions would be formulated such that they would contain an effective dose of the compound of the invention. In general, solid dosage forms containing 100 mg-1500 mg of the compound of the invention are contemplated. Preparations suitable for oral suspension would contain a similar dosage.
Pharmaceutical formulations can be formulated together with auxiliaries and additives

usually employed in pharmacy, such as tablet binders, fillers, preservatives, tablet disintegrating agents, flow regulating, agents, plasticizers, wetting agents, dispersing agents, emulsifiers, solvents, pH altering additives, flavourings and the like. A second preferred method is parenterally for intramuscular, intravenous or subcutaneous administration.
When the pharmaceutical composition is formulated into an injectable preparation, in formulating the pharmaceutical composition into the form of a solution or suspension, all diluents customarily used in the art can be used. Examples of suitable diluents are water, ethyl alcohol, polypropylene glycol, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol, and sorbitan esters. Sodium chloride, glucose or glycerol may be incorporated into a therapeutic agent. It is preferred that the concentration of active ingredient in the injectable preparation be in the range of 0.1 mg/ml to 100 mg/ml.
In addition to the common dosage forms set out above, the compounds of the present invention may also be administered by controlled release means and/or delivery devices such as those described in U.S. Patent No. 3,845,770; U.S. Patent No 3,916,899; U.S. Patent No 3,536,809; U.S. Patent No 3,598,123 and U.S. Patent No 4,008,719; the disclosures of which are hereby incorporated by reference.
The total daily dose range is generally from about 200 mg to about 5000 mg of the polymorphs of the invention. However, the dose may be higher or lower depending on the needs and conditions of the patient.
The polymorphs and the pharmaceutical compositions of the invention are useful in the treatment of patient having a broad spectrum of bacterial diseases caused by infections such as impetigo, pneumonia, bronchitis, pharyngitis, endocarditis, urinary tract infections, gastro-intestinal infections and bacteremia. These bacterial infections could be caused by any of the following bacteria - Staphylococcus aureus, coagulase negative staphylococci, methicillin-resitant Staphylococcus aureus, methicillin-resitant coagulase negative staphylococci, enterococci, beta-haemolytic streptococci, viridans group of

streptococci, mycobacterial infections due to multi-drug resistant M. tuberculosis and other atypical mycobacteria such as M. intracellulare and M. avium, as well as newly emerging Gram-negative pathogens such as Chryseobacterium meningosepticum, Chryseobacterium indologense and other Gram-negative pathogens such as E. coli, Klebsiella, Proteus, Serratia, Citrobacter, and Pseudomonas.
The invention also encompasses an anti infective composition for the treatment of patient in need of prophylaxis and/or therapy for systemic infections especially resistant gram-positive organism infections, gram-negative organism infections, mycobacterial infections and nosocomial pathogen infections, which composition comprises an amount of the compound of the invention substantially sufficient to eradicate said infection. The compound and compositions of this invention can be administered to patient that are at risk of being infected, for example a compound or composition of this invention can be administered to a patient prior to and/or after surgery, health care workers or others who are at risk of being infected.
The invention also encompasses administering the compounds to a patient. The compound and compositions to be used in the invention must, accordingly, be pharmaceutically acceptable. As used herein, such a "pharmaceutically acceptable" component is one that is suitable for use with humans and/or animals without undue adverse side effects (such as toxicity, irritation, and allergic response) commensurate with a reasonable benefit/risk ratio.
The term patient as used herein is taken to mean birds, fishes, domestic animals and mammals, for example, humans, cats, dogs, horses, sheep, bovine cows, pigs, lambs, rats, mice and guinea pigs.
The following detailed examples serve to more fully illustrate the invention without limiting its scope. It is understood that various other embodiments and modifications in the practice of the invention will be apparent to, and can be readily made by, those ordinary skill in the art without departing from the scope and spirit of the invention as

described above. Accordingly, it is not intended that the scope of the claims appended hereto be limited to the exact description set forth above, but rather than the claims be construed as encompassing all of the features of patentable novelty that reside in the present invention, including all of the features and embodiments that would be treated as equivalents thereof by those skilled in the relevant art. The invention is further described with reference to the following experimental work.
The following detailed examples serve to more fully illustrate the invention without limiting its scope.
Examples
Preparation of crystalline polymorph of S-(-)-9-fluoro-6,7-dihydro-8-(4-L-
alaninyIoxypiperidin-l-yl)-5-methyl-l-oxo-lH,5H-benzo[ij] quinolizine-2-
carboxylic acid methanesulfonic acid salt
To a mixture of (2'S,5S)-9-fluoro-6,7-dihydro-8-{4-N-tert-butoxycarbonyl-L-alaninyloxy-piperidin-1 -yl} -5-methyl-1 -oxo-1 H,5H-benzo[i j]quinolizine-2-carboxylic acid (415 g) in acetone (4.5 L) was charged methanesulfonic acid (66 ml). Reaction mixture was stirred at 65-67 °C temperature for overnight. The suspension was filtered at 40-45 °C. Solid was washed with acetone (1.5 L) to provide an off white solid, which was dried under 40 to 45 mm vacuum at 55-60 °C temperature over the period of 3-4 h to provide crude material 383.0 g quantity. The crude material was dissolved under stirring in a water (2.4 Ltr) to obtain slightly hazy solution. The hazy solution was washed with ethyl acetate (250 ml X 3) to remove impurities. The aqueous layer was filtered over a Inflow bed. The clear aqueous filtrate was concentrated under vacuum to dryness by keeping temperature below 60°C to provide a solid mass. The solid mass was weighed and equal volume of water was added to it under stirring, followed by acetone (volume 12 times of the weight of solid mass). The suspension was stirred for 15 to 18 hrs at 30-35° C. The suspension as filtered, the wet cake was washed with acetone and was dried in oven under vacuum at 40-45°C to provide crystalline polymorph of the invention in 330 g (80%) quantity.
MF: C23H30FN3O8S, MS (ES+) m/z 432 (obtained as free base for MF: C22H26FN3O5); M.P. 278.50 °C by DSC

Infra-red spectrum (cm-1): 3429, 2957, 1739, 1619, 1523, 1442, 1390, 1322, 1273, 1228,
1149,1071, 1039,883,810.
X-ray powder diffraction (2G): 9.82 ± 0.2°, 11.76 ± 0.2°, 12.66 + 0.2°, 13.38 ± 0.2°,
15.56 ± 0.2°, 16.34 ± 0.2°, 17.58 ± 0.2°, 18.26 ± 0.2°, 19.74 ± 0.2°, 20.86 ± 0.2°, 21.70 ±
0.2°, 22.64 ± 0.2°, 23.58 ± 0.2°, 28.38 ± 0.2°, 33.18 ± 0.2°.
DSC endotherm at 278.50 °C (onset at 275.92 °C).
Preparation of amorphous polymorph of S-(-)-9-fluoro-6,7-dihydro-8-(4-L-
alaninyloxypiperidin-l-yl)-5-methyl-l-oxo-lH,5H-benzo[i,j] quinoIizine-2-
carboxylic acid methanesulfonic acid salt
Crystalline polymorph of S-(-)-9-fluoro-6,7-dihydro-8-(4-L-alaninyloxypiperidin-l-yl)-5-
methyl-l-oxo-lH,5H-benzo[ij] quinolizine-2-carboxylic acid methanesulfonic acid salt
(10 g) was dissolved in water (50 ml) and was lyophilized at freeze drying temperature.
Free flowing off white material in 10 g quantity was obtained as an amorphous
polymorph.
MF: C23H30FN3O8S, MS (ES+) m/z 432 (obtained as free base for MF: C22H26FN3O5);
m.p. 272.00 °C by DSC.
Infra-red spectrum (cm"1): 3446, 2959, 1738, 1621, 1528, 1451, 1393, 1327, 1205, 1047,
882, 810.
X-ray powder diffraction (29): 19.86 ± 0.2°, 20.02 ± 0.2°, 20.12 ± 0.2°, 20.28 ± 0.2°,
20.36 + 0.2°, 20.74 ± 0.2°, 21.14 ± 0.2°, 25.36 ± 0.2°, 25.76 ± 0.2°, 25.94 ± 0.2°, 26.14 ±
0.2°, 26.28 ± 0.2°.
DSC endotherm at 44.50 °C (onset at 42.50 °C), endotherm at 160.50 °C (onset at 154.33
°C), exotherm at 202.66 °C (onset at 190.53 °C), endotherm at 272.00 °C (onset at
267.97 °C).
Test Example-1
X-ray Powder Diffraction Analysis: 300 mg each of the teat sample prepared as above were thinly spread on a sample holder. X-ray powder diffraction analyses (40kv x 40 mA

Rigaku D/max 2200) were performed under the conditions listed below:
Scan speed 5°/ min
Sampling time 7 min
Scan mode: continuous
29/0 reflection
Cu target (Ni filter) Results of the X-ray diffraction analysis on the samples amorphous forms are depicted. From these spectra it can be verified that crystal forms differ from each other.
Test Example-2
Thermal Analysis: For the Differential Scanning Calorimetry, PERKIN-ELMER system was used. 5.4 mg of the sample was weighed into the aluminum pan, which was then press sealed with an aluminium lid. After three tiny needle holes were made on the lid the sample was tested by heating from 30°C to 300 °C at a rate of 10 °C/min.
Test Example-3
Infra-red spectrum analysis: Infra-red spectrum was obtained on BRUCKER VECTOR 22 system and by using KBr pellet.
Claim:
1. A crystalline polymorph of S-(-)-9-fluoro-6,7-dihydro-8-(4-L-alaninyloxypiperidin-1 -yl)-5-methyl-1 -oxo-1 H,5H-benzo[i j] quinolizine-2-carboxylic acid methanesulfonic acid salt having an X-ray diffraction containing the 26 value at 9.82 ± 0.2°, 18.26 ± 0.2° and 19.74 ± 0.2.
2. The crystalline polymorph as claimed in claim 1 having an X-ray diffraction containing the 20 value at 9.82 ± 0.2°, 11.76 ± 0.2°, 12.66 ± 0.2°, 13.38 ± 0.2°, 15.56 ± 0.2°, 16.34 ± 0.2°, 17.58 ± 0.2°, 18.26 ± 0.2°, 19.74 ± 0.2°, 20.86 ± 0.2°, 21.70 ± 0.2°, 22.64 ± 0.2°, 23.58 ± 0.2°, 28.38 ± 0.2°, 33.18 ± 0.2°.
3. A amorphous polymorph of S-(-)-9-fluoro-6,7-dihydro-8-(4-L-alaninyloxypiperidin-1 -yl)-5-methyl-1 -oxo-1 H,5H-benzo[i j] quinolizine-2-carboxylic acid methanesulfonic acid salt having an X-ray diffraction containing the 20 value at 19.86 + 0.2°, 20.02 + 0.2°, 20.12 ± 0.2°, 20.28 ± 0.2°, 20.36 ± 0.2°, 20.74 + 0.2°, 21.14 ± 0.2°, 25.36 + 0.2°, 25.76 ± 0.2°, 25.94 ± 0.2°, 26.14 ± 0.2°, 26.28 ± 0.2°.
4. A process for the preparation of crystalline polymorph of S-(-)-9-fluoro-6,7-dihydro-8-(4-L-alaninyloxypiperidin-1 -yl)-5-methyl-1 -oxo-1 H,5H-benzo[i j] quinolizine-2-carboxylic acid methanesulfonic acid salt, comprising the steps of:
a) reacting (2'S,5S)-9-fluoro-6,7-dihydro-8-{4-N-tert-butoxycarbonyl-L-alaninyloxy-piperidin-1 -yl} -5-methyl-1 -oxo- lH,5H-benzo[i jjquinolizine-2-carboxylic acid with methanesulfonic acid in ketone solvent to get crude S-(-)-9-fluoro-6,7-dihydro-8-(4-L-alaninyloxypiperidin-1 -yl)-5-methyl-1 -oxo-lH,5H-benzo[i,j] quinolizine-2-carboxylic acid methanesulfonic acid salt,

b) contacting the crude S-(-)-9-fluoro-6,7-dihydro-8-(4-L-alaninyloxypiperidin-1 -yl)-5-methyl-1 -oxo-1 H,5H-benzo[i j ] quinolizine-2-carboxylic acid methanesulfonic acid salt with water,
c) treating the mass with ketone solvent and
d) isolating the crystalline polymorph of S-(-)-9-fluoro-6,7-dihydro-8-(4-L-alaninyloxypiperidin-1 -yl)-5-methyl-1 -oxo-1 H,5H-benzo[ij] quinolizine-2-carboxylic acid methanesulfonic acid salt.
5. A process of preparation of amorphous polymorph of S-(-)-9-fluoro-6,7-dihydro-
8-(4-L-alaninyloxypiperidin-1 -yl)-5-methyl- 1-oxo-1 H,5H-benzo[i j]quinolizine-
2-carboxylic acid methanesulfonic acid salt, comprising the steps of;
a) dissolving the crystalline polymorph of S-(-)-9-fluoro-6,7-dihydro-8-(4-L-alaninyloxypiperidin-l-yl)-5-methyl-l-oxo-lH,5H-benzo[ij] quinolizine-2-carboxylic acid methanesulfonic acid salt with water
b) lyophilizing at freeze drying temperature and
c) isolated the amorphous polymorph of S-(-)-9-fluoro-6,7-dihydro-8-(4-L-alaninyloxypiperidin-1 -yl)-5-methyl-1 -oxo-1H, 5H-benzo[ij ]quinolizine-2-carboxylic acid methanesulfonic acid salt.

6. A pharmaceutical composition, which comprise crystalline polymorph of S-(-)-9-fluoro-6,7-dihydro-8-(4-L-alaninyloxypiperidin-1 -yl)-5-methyl-1 -oxo-1 H,5H-benzo[ij]quinolizine-2-carboxylic acid methanesulfonic acid salt and a pharmaceutically acceptable carrier.
7. A pharmaceutical composition, which comprise amorphous polymorph of S-(-)-9-fluoro-6,7-dihydro-8-(4-L-alaninyloxypiperidin-1 -yl)-5-methyl-1 -oxo-1 H,5H-benzo[ij]quinolizine-2-carboxylic acid methanesulfonic acid salt and pharmaceutically acceptable carrier.
8. The pharmaceutical composition as claimed in claim 6 or 7 comprises injectable solutions, suspensions, emulsions, tablets, coated tablets, coated tablet cores,

capsules, solutions, troches, dispersions, pellets, granules, suppositories, hard or soft gelatin capsules.
9. A method for treating bacterial diseases in a patient comprising administering an effective amount of crystalline polymorph of S-(-)-9-fluoro-6,7-dihydro-8-(4-L-alaninyloxypiperidin-1 -yl)-5-methyl-1 -oxo-1 H,5H-benzo[ij]quinolizine-2-carboxylic acid methanesulfonic acid salt.
10. A method for treating bacterial diseases in a patient comprising administering an effective amount of amorphous polymorph of S-(-)-9-fluoro-6,7-dihydro-8-(4-L-alaninyloxypiperidin-1 -yl)-5-methyl-1 -oxo-1 H,5H-benzo[i j] quinolizine-2-carboxylic acid methanesulfonic acid salt.


Abstract
The invention relates to crystalline and amorphous polymorphs of S-(-)-9-fiuoro-6,7-dihydro-8-(4-L-alaninyloxypiperidin-1 -yl)-5-methyl-1 -oxo-1 H,5H-benzo[ij ] quinolizine-2-carboxylic acid methanesulfonic acid salt and its pharmaceutical compositions. The polymorphs and compositions of the invention are useful to treat bacterial Gram-positive, Gram-negative and anaerobic infections, especially infections caused by resistant Gram-positive organism and Gram-negative organism, mycobacterial infections and emerging nosocomial pathogen infections.