U.S. patent application number 12/318845 was filed with the patent office on 2009-07-16 for salt of naphthyridine carboxylic acid derivative.
This patent application is currently assigned to LG Life Sciences, Ltd.. Invention is credited to Jay Hyok Chang, Hoon Choi, Jong Ryoo Choi, Ae Ri Kim, Jin Hwa Lee, Tae Hee Lee, Do Hyun Nam, Ki Sook Park.
Application Number | 20090182006 12/318845 |
Document ID | / |
Family ID | 19500439 |
Filed Date | 2009-07-16 |
United States Patent
Application |
20090182006 |
Kind Code |
A1 |
Kim; Ae Ri ; et al. |
July 16, 2009 |
Salt of naphthyridine carboxylic acid derivative
Abstract
7-(3-Aminomethyl-4-methoxyiminopyrrolidin-1-yl)-1-cyclopropyl-6-fluoro-4--
oxo-1,4-dihydro-1,8-naphthyridine-3-carboxylic acid
methanesulfonate and hydrates thereof, processes for their
preparation, pharmaceutical compositions comprising them, and their
use in antibacterial therapy.
Inventors: |
Kim; Ae Ri; (Daejeon,
KR) ; Lee; Jin Hwa; (Daejeon, KR) ; Park; Ki
Sook; (Daejeon, KR) ; Choi; Jong Ryoo;
(Daejeon, KR) ; Lee; Tae Hee; (Daejeon, KR)
; Chang; Jay Hyok; (Daejeon, KR) ; Nam; Do
Hyun; (Daejeon, KR) ; Choi; Hoon; (Daejeon,
KR) |
Correspondence
Address: |
FINNEGAN, HENDERSON, FARABOW, GARRETT & DUNNER;LLP
901 NEW YORK AVENUE, NW
WASHINGTON
DC
20001-4413
US
|
Assignee: |
LG Life Sciences, Ltd.
|
Family ID: |
19500439 |
Appl. No.: |
12/318845 |
Filed: |
January 9, 2009 |
Related U.S. Patent Documents
|
|
|
|
|
|
Application
Number |
Filing Date |
Patent Number |
|
|
11635069 |
Dec 7, 2006 |
|
|
|
12318845 |
|
|
|
|
11402048 |
Apr 12, 2006 |
|
|
|
11635069 |
|
|
|
|
10824609 |
Apr 15, 2004 |
|
|
|
11402048 |
|
|
|
|
10223850 |
Aug 20, 2002 |
6723734 |
|
|
10824609 |
|
|
|
|
09923580 |
Aug 6, 2001 |
|
|
|
10223850 |
|
|
|
|
09381491 |
Dec 8, 1999 |
|
|
|
PCT/KR98/00051 |
Mar 20, 1998 |
|
|
|
09923580 |
|
|
|
|
Current U.S.
Class: |
514/300 ;
546/123 |
Current CPC
Class: |
C07D 471/04 20130101;
A61K 31/4375 20130101; A61P 31/04 20180101; A61P 13/02 20180101;
C07B 2200/13 20130101; A61P 11/16 20180101; C07C 309/04
20130101 |
Class at
Publication: |
514/300 ;
546/123 |
International
Class: |
A61K 31/4375 20060101
A61K031/4375; C07D 471/04 20060101 C07D471/04; A61P 31/04 20060101
A61P031/04 |
Foreign Application Data
Date |
Code |
Application Number |
Mar 21, 1997 |
KR |
1997/9840 |
Claims
1.
7-(3-Aminomethyl-4-methoxyiminopyrrolidin-1-yl)-1-cyclopropyl-6-fluoro-
-4-oxo-1,4-dihydro-1,8-naphthyridine-3-carboxylic acid
methanesulfonate.
2.
7-(3-Aminomethyl-4-methoxyiminopyrrolidin-1-yl)-1-cyclopropyl-6-fluoro-
-4-oxo-1,4-dihydro-1,8-naphthyridine-3-carboxylic acid
methanesulfonate.nH.sub.2O, wherein n is in the range of from 1 to
4.
3. A compound according to claim 2 wherein n is 1.5.
4. A compound according to claim 2 having peaks at 2.theta.=8.0,
12.2 and 14.70 in its X-ray diffraction pattern.
5. A compound according to claim 2 having an X-ray diffraction
pattern substantially as shown in FIG. 7.
6. A compound according to claim 2 wherein n is 3.
7. A compound according to claim 2 having peaks at 2.theta.=7.7 and
11.8.degree. in its X-ray diffraction pattern.
8. A compound according to claim 2 having an X-ray diffraction
pattern substantially as shown in FIG. 6.
9. A compound according to claim 2 which has a moisture content of
from 4 to 6%.
10. A compound according to claim 2 which has a moisture content of
from 9 to 11%.
11. A pharmaceutical composition comprising a compound according to
any one of the preceding claims, together with a pharmaceutically
acceptable carrier or excipient.
12. A compound according to any one of claims 1 to 10, for use as a
pharmaceutical.
13. A method of treating bacterial infections in humans and animals
which comprises administering a therapeutically effective amount of
a compound according to any one of claims 1 to 10.
14. The use of a compound according to any one of claims 1 to 10
for the manufacture of a medicament for treating bacterial
infection.
15. A process for the preparation of a compound according to any
one of claims 1 to 10, which comprises reacting
7-(3-aminomethyl-4-methoxyiminopyrrolidin-1-yl)-1-cyclopropyl-6-fluoro-4--
oxo-1,4-dihydro-1,8-naphthyridine-3-carboxylic acid with
methanesulfonic acid and crystallizing the resulting compound from
solution, and where desired or necessary, adjusting the hydration
of the compound.
16. A process for the preparation of a compound according to any
one of claims 2 to 10, comprising exposing
7-(3-aminomethyl-4-methoxyiminopyrrolidin-1-yl)-1-cyclopropyl-6-fluoro-4--
oxo-1,4-dihydro-1,8-naphthyridine-3-carboxylic acid
methanesulfonate anhydrate or a solvate thereof to a relative
humidity of at least 75%.
17. A process according to claim 16, wherein the solvate is a
solvate with one or more organic solvents selected from
C.sub.1-C.sub.4 haloalkanes and alcohols.
18. A solvate of
7-(3-aminomethyl-4-methoxyiminopyrrolidin-1-yl)-1-cyclopropyl-6-fluoro-4--
oxo-1,4-dihydro-1,8-naphthyridine-3-carboxylic acid
methanesulfonate with one or more organic solvents.
Description
TECHNICAL FIELD
[0001] The present invention relates to a salt and associated
hydrates of racemic
7-(3-aminomethyl-4-methoxyiminopyrrolidin-1-yl)-1-cyclopropyl-6-f-
luoro-4-oxo-1,4-dihydro-1,8-naphthyridine-3-carboxylic acid,
processes for their preparation, pharmaceutical compositions
comprising them, and their use in antibacterial therapy.
BACKGROUND ART
[0002] EP 688772 (corresponding to Korean Patent Laid open
Publication No 96-874) discloses novel
quinoline(naphthyridine)carboxylic acid derivatives, including
anhydrous
7-(3-aminomethyl-4-methoxyiminopyrrolidin-1-yl)-1-cyclopropyl-6-fluoro-4--
oxo-1,4-dihydro-1,8-naphthyridine-3-carboxylic acid of formula I,
having antibacterial activity.
##STR00001##
DISCLOSURE OF INVENTION
[0003] According to the invention there is provided
7-(3-Aminomethyl-4-methoxyiminopyrrolidin-1-yl)-1-cyclopropyl-6-fluoro-4--
oxo-1,4-dihydro-1,8-naphthyridine-3-carboxylic acid
methanesulfonate.
[0004]
7-(3-Aminomethyl-4-methoxyiminopyrrolidin-1-yl)-1-cyclopropyl-6-flu-
oro-4-oxo-1,4-dihydro-1,8-naphthyridine-3-carboxylic acid
methanesulfonate (hereinafter referred to as `the
methanesulfonate`) may be obtained as an anhydrate or a hydrate
i.e. methanesulfonate.nH.sub.2O.
[0005] Hydrates of the methanesulfonate wherein n is in the range 1
to 4 are preferred. Particular hydrates of the methanesulfonate
which may be mentioned are those in which n is 1, 1.5, 2, 2.5, 3,
3.5 and 4. Particularly preferred compounds are those in which n is
1.5 or 3, with n=1.5 being most preferred.
[0006] The moisture content of the methanesulfonate hydrates varies
with the hydration number (n) of the hydrated molecule. The
methanesulfonate has a molecular weight of 485.5 thus the
calculated moisture content of hydrates where n is 1, 1.5, 2, 2.5,
3, 3.5 and 4 is 3.6%, 5.0%, 6.9%, 8.5%, 10.0%, 11.5% and 12.9%
respectively. However, the actual moisture content of the
methanesulfonate hydrates may differ from the calculated value
depending on various factors including recrystallization conditions
and drying conditions. The observed moisture content for the
methanesulfonate hydrates where n is 1, 1.5, 2, 2.5, 3, 3.5 and 4
is shown in Table 1:
TABLE-US-00001 TABLE 1 Hydration Number (n) Moisture Content (%
w/w) 1 2~4 1.5 4~6 2 6~8 2.5 8~9 3 9~11 3.5 11~12 4 12~13
[0007] It is possible to mix methanesulfonate hydrates having
different moisture contents together to give materials having
intermediate moisture contents.
[0008] Preferred methanesulfonate hydrates have a moisture content
of from 4 to 6% or from 9 to 11%, especially a moisture content of
from 4 to 6%.
[0009] The methanesulfonate has been observed to exist as a stable
hydrate over a range of hydration numbers (n). Stability of the
hydrate refers to its resistance to loss or gain of water molecules
contained in the compound. The methanesulfonate hydrates maintain a
constant moisture content over an extended relative humidity range.
The n=3 hydrate has a constant moisture content at a relative
humidity of from at least 23 to 75%, and the n=1.5 hydrate has a
constant moisture content at a relative humidity of from 23 to 64%
(see FIGS. 3 and 4). In contrast, moisture absorption by the
anhydrate varies greatly with relative humidity.
[0010] Both the methanesulfonate anhydrate and n=3 hydrate undergo
transition to the n=1.5 hydrate in aqueous suspension indicating
that the latter is thermodynamically more stable. The n=1.5 hydrate
is a sesquihydrate at 11 to 64% of relative humidity. Above 75%
relative humidity, it takes up water over 10% and its X-ray
diffraction pattern changes. The hydrate (another form of n=3
having different physiochemical properties from the n=3 hydrate of
Example 2) obtained from n=1.5 hydrate at 93% relative humidity is
not stable at lower relative humidity, it converts back to n=1.5
hydrate at a relative humidity below 75%.
[0011] Since the moisture content of the anhydrate changes readily
depending on the environment e.g. relative humidity, formulation
additives etc, it may require careful handling during storage or
formulation, with operations such as quantifying procedures being
performed in a dry room. The hydrates do not change in moisture
content as easily and hence products which are stable to prolonged
storage and formulation may be obtained. The hydrate can be
tabletted without the addition of a binder since the water
contained in the compound itself acts a binder, whereas it may not
be possible to tablet the anhydrate at a similar pressure.
[0012] The present invention also provides a process for the
preparation of
7-(3-aminomethyl-4-methoxyiminopyrrolidin-1-yl)-1-cyclopropyl-6-fluoro-
-4-oxo-1,4-dihydro-1,8-naphthyridine-3-carboxylic acid
methanesulfonate and hydrates thereof which comprises reacting
7-(3-aminomethyl-4-methoxyiminopyrrolidin-1-yl)-1-cyclopropyl-6-fluoro-4--
oxo-1,4-dihydro-1,8-naphthyridine-3-carboxylic acid with
methanesulfonic acid and crystallizing the resulting
methanesulfonate from solution, and where desired or necessary
adjusting the hydration of the compound.
[0013] The methanesulfonate and its hydrates may be prepared by the
addition of methanesulfonic acid to the free base which may be
prepared as described in EP 688772. Preferably, 0.95 to 1.5 molar
equivalents of methanesulfonic acid is added to the free base, or 1
molar equivalent of methanesulfonic acid dissolved in a suitable
solvent is added to the free base. Suitable solvents for the
preparation of the methanesulfonate and its hydrates include any
solvent in which the methanesulfonate is substantially insoluble,
suitable solvents include C.sub.1-C.sub.4 haloalkanes,
C.sub.1-C.sub.8 alcohols and water, or mixtures thereof.
Dichloromethane, chloroform, 1,2-dichloroethane, methanol, ethanol,
propanol and water, or mixtures thereof, are preferred solvents. If
necessary, the free base may be heated in the solvent to facilitate
solution before methanesulfonic acid is added, alternatively the
methanesulfonic acid may be added to a suspension, or partial
suspension, of the free base in the solvent. Following addition of
the methanesulfonic acid the reaction mixture is preferably allowed
to stand or is stirred for 1 to 24 hours at a temperature of from
about -10 to 40.degree. C. The resulting methanesulfonate is
obtained as a solid which can be isolated by filtration or by
removal of the solvent under reduced pressure.
[0014] Different hydrates may be obtained by altering the
recrystallization conditions used in the preparation of the
methanesulfonate, such conditions may be ascertained by
conventional methods known to those skilled in the art.
[0015] The present invention also provides a process for the
preparation of a hydrate of
7-(3-aminomethyl-4-methoxyiminopyrrolidin-1-yl)-1-cyclopropyl-6-fluoro-4--
oxo-1,4-dihydro-1,8-naphthyridine-3-carboxylic acid
methanesulfonate comprising exposing the methanesulfonate anhydrate
or a solvate thereof to a high relative humidity.
[0016] The methanesulfonate anhydrate or solvate thereof is
preferably exposed to a relative humidity of at least 75%.
[0017] The methanesulfonate anhydrate or solvate thereof may be
exposed to high relative humidity by passing humidified nitrogen
gas through the methanesulfonate anhydrate or solvate thereof or by
standing the methanesulfonate anhydrate or solvates thereof under a
high relative humidity.
[0018] The humidified nitrogen gas used in this process, for
example nitrogen gas having a humidity of at least 75%, may be made
by conventional methods. In this process it is desirable to
maintain the temperature in the range above which moisture
condensation could occur. Also, particularly in large scale
production, it is preferable to stir the sample thoroughly while
the humidified nitrogen gas is passed through. When the hydrate is
prepared by standing the methanesulfonate anhydrate or solvate
thereof under a high relative humidity, for example a relative
humidity of at least 75%, it is preferable to spread the sample as
thinly as possible in order to raise the conversion efficiency.
[0019] The solvates of methanesulfonate anhydrate which may be used
in the process according to this aspect of the present invention
include solvates with one or more organic solvents. Preferred
solvents include C.sub.1-C.sub.4 haloalkanes and C.sub.1-C.sub.8
alcohols, for example those selected from the group consisting of
ethanol, dichloromethane, isopropanol and 2-methyl-2-propanol.
[0020] Solvates of the methanesulfonate anhydrate are novel. Thus
according to a further aspect of the invention there is provided a
solvate of
7-(3-aminomethyl-4-methoxyiminopyrrolidin-1-yl)-1-cyclopropyl-6-fluoro-4--
oxo-1,4-dihydro-1,8-naphthyridine-3-carboxylic acid
methanesulfonate with one or more organic solvents.
[0021] The solvates of the methanesulfonate are prepared by
recrystallization and controlled by the condition of
recrystallizing system.
[0022] The methanesulfonate and its hydrates exhibit the same
potent antibacterial activity as the corresponding free base
disclosed in EP 688772. The methanesulfonate and its hydrates also
exhibit desirable physicochemical properties including improved
solubility and constant moisture content regardless of the ambient
relative humidity when compared to the free base and other salts
thereof. The methanesulfonate and its hydrates thus exhibit greater
ease of handling, quality control and formulation than the free
base and other salts thereof.
[0023] As mentioned above the methanesulfonate and its hydrates
exhibit antibacterial activity. The methanesulfonate and its
hydrates may be formulated for administration in any convenient way
for use in human or veterinary medicine, according to techniques
and procedures per se known in the art with reference to other
antibiotics, and the invention therefore includes within its scope
a pharmaceutical composition comprising
7-(3-aminomethyl-4-methoxyiminopyrrolidin-1-yl)-1-cyclopropyl-6-fluoro-4--
oxo-1,4-dihydro-1,8-naphthyridine-3-carboxylic acid
methanesulfonate or a hydrate thereof together with a
pharmaceutically acceptable carrier or excipient.
[0024] Compositions comprising the methanesulfonate or hydrate
thereof as active ingredient may be formulated for administration
by any suitable route, such as oral, parenteral or topical
application. The compositions may be in the form of tablets,
capsules, powders, granules, lozenges, creams or liquid
preparations, such as oral or sterile parenteral solutions or
suspensions. Tablets and capsules for oral administration may be in
unit dose presentation form and may contain conventional excipients
such as binding agents, for example, hydroxypropyl methyl
cellulose, hydroxy propyl cellulose, syrup acacia, gelatin,
sorbitol, tragacanth, or polyvinylpyrrollidone; fillers, for
example microcrystalline cellulose, lactose, sugar, maize-starch,
calcium phosphate, sorbitol or glycine; tabletting lubricants, for
example magnesium stearate, talc, polyethylene glycol or silica;
disintegrants, for example sodium starch glycolate, cross linked
polyvinyl pyrollidone or potato starch; or acceptable wetting
agents such as sodium lauryl sulfate. The tablets may be coated
according to methods well known in normal pharmaceutical practice.
Oral liquid preparations may be in the form of, for example,
aqueous or oily suspensions, solutions, emulsions, syrups or
elixirs, or may be presented as a dry product for reconstitution
with water or other suitable vehicle before use. Such liquid
preparations may contain conventional additives such as suspending
agents, for example sorbitol, methyl cellulose, glucose syrup,
gelatin, hydroxyethyl cellulose, caboxymethyl cellulose, aluminium
stearate gel or hydrogenated edible fats; emulsifying agents, for
example lecithin, sorbitan monooleate, or acacia; non-aqueous
vehicles (which may include edible oils), for example almond oil,
oily esters, glycerine, propylene glycol, or ethyl alcohol;
preservatives, for example methyl or propyl p-hydroxybenzoate or
sorbic acid; and, if desired conventional flavouring or coloring
agents. Suppositories will contain conventional suppository base,
e.g. cocoa-butter or other glyceride.
[0025] For parenteral administration, fluid unit dosage forms are
prepared utilising the compound and a sterile vehicle, water being
preferred. The methanesulfonate or hydrate thereof, can be either
suspended or dissolved in the vehicle, depending on the vehicle and
concentration used. In preparing solutions the methanesulfonate or
hydrate thereof can be dissolved in water for injection and filter
sterilized before filling into a suitable vial or ampoule and
sealing. Advantageously, agents such as local anaesthetic,
preservative and buffering agents can be dissolved in the vehicle.
To enhance the stability, the composition can be lyophilised and
the dry lyophilised powder sealed in a vial, an accompanying vial
of water for injection may be supplied to reconstitute the powder
prior to use. Parenteral suspensions are prepared in substantially
the same manner except that the methanesulfonate or hydrate thereof
is suspended in the vehicle instead of being dissolved and
sterilization cannot be accomplished by filtration. The
methansulfonate or hydrate thereof can be sterilized by exposure to
ethylene oxide before suspending in the sterile vehicle.
Advantageously, a surfactant or wetting agent is included in the
composition to facilitate uniform distribution of the
methanesulfonate or hydrate thereof.
[0026] The methanesulfonate or hydrate thereof may also be
formulated as an intramammary composition for veterinary use.
[0027] The composition may contain from 0.1% to 100% by weight,
preferably from 10 to 99.5% by weight, more preferably from 50 to
99.5% by weight of the active ingredient measured as the free base,
depending on the method of administration. Where the compositions
comprise dosage units, each unit will preferably contain from
50-1500 mg of the active ingredient measured as the free base. The
dosage as employed for adult human treatment will preferably range
from 100 mg to 12 g per day for an average adult patient (body
weight 70 kg), for instance 1500 mg per day, depending on the route
and frequency of administration. Such dosages correspond to
approximately 1.5 to 170 mg/kg per day. Suitably the dosage is from
1 to 6 g per day.
[0028] The daily dosage is suitably given by administering the
active ingredient once or several times in a 24-hour period, e.g.
up to 400 mg maybe adminstered once a day, in practice, the dosage
and frequency of administration which will be most suitable for an
individual patient will vary with the age, weight and response of
the patients, and there will be occasions when the physician will
choose a higher or lower dosage and a different frequency of
administration. Such dosage regimens are within the scope of this
invention.
[0029] The present invention also includes a method of treating
bacterial infections in humans and animals which method comprises
administering a therapeutically effective amount of
7-(3-aminomethyl-4-methoxyiminopyrrolidin-1-yl)-1-cyclopropyl-6-fluoro-4--
oxo-1,4-dihydro-1,8-naphthyridine-3-carboxylic acid
methanesulfonate or a hydrate thereof.
[0030] In a further aspect, the present invention also provides the
use of
7-(3-aminomethyl-4-methoxyiminopyrrolidin-1-yl)-1-cyclopropyl-6-fluoro-4--
oxo-1,4-dihydro-1,8-naphthyridine-3-carboxylic acid
methanesulfonate or a hydrate thereof for the manufacture of a
medicament for treating bacterial infection.
[0031] The methanesulfonate and its hydrates are active against a
broad range of Gram-positive and Gram-negative bacteria, and may be
used to treat a wide range of bacterial infections including those
in immunocompromised patients.
[0032] Amongst many other uses, the methanesulfonate and its
hydrates are of value in the treatment of skin, soft tissue,
respiratory tract and urinary tract infections, and sexually
transmitted diseases in humans. The methanesulfonate and its
hydrates may also be used in the treatment of bacterial infections
in animals, such as mastitis in cattle.
BRIEF DESCRIPTION OF DRAWINGS
[0033] The following examples and figures illustrate the invention
but are not intended to limit the scope in any way.
[0034] FIG. 1 shows the moisture sorption profile of
methanesulfonate anhydrate of Example 1 at 25.degree. C. at several
relative humidities.
[0035] FIG. 2 shows the isothermal moisture sorption profile of
methanesulfonate anhydrate of Example 1 at 25.degree. C.
[0036] FIG. 3 shows the equilibrium moisture content of the
methanesulfonate n=3 hydrate of Example 2 at a relative humidity of
23 to 75%.
[0037] FIG. 4 shows the equilibrium moisture content of the
methanesulfonate n=1.5 hydrate of Example 3 at a relative humidity
of 23 to 75%.
[0038] FIG. 5 shows the powder X-ray diffraction pattern of the
methanesulfonate anhydrate of Example 1.
[0039] FIG. 6 shows the powder X-ray diffraction pattern of the
methanesulfonate n=3 hydrate of Example 2. The characteristic peaks
are 2.theta.=7.7, 11.8.degree.. The exact position of peaks can
vary slightly depending on the experimental conditions.
[0040] FIG. 7 shows the powder X-ray diffraction pattern of the
methanesulfonate n=1.5 hydrate of Example 3. The characteristic
peaks are 2.theta.=8.0, 12.2, 14.7.degree.. The exact position of
peaks can vary slightly depending on the experimental
conditions.
[0041] FIG. 8 shows the variation in moisture content with elapsed
time of the methanesulfonate anhydrate of Example 1, taken after 0,
5, 10, 20, 30, and 60 minutes, respectively, from the initial point
of passing humidified nitrogen gas through;
[0042] FIG. 9 shows the Differential Scanning Calorimetry on the
methanesulfonate anhydrate of Example 1 and the methanesulfonate
n=3 hydrate of Example 2.
[0043] FIG. 10 shows the results of thermogravimetric analysis on
the methanesulfonate n=3 hydrate of Example 2.
[0044] FIG. 11 shows the change in X-ray diffraction pattern with
elapsed time of the methanesulfonate solvate (ethanol content
0.11%) of Example 4, from initial point of passing the humidified
nitrogen gas having a relative humidity of 93% through.
[0045] FIG. 12 shows the change in X-ray diffraction pattern with
elapsed time of the methanesulfonate solvate (ethanol content 1.9%)
of Example 5, from the initial point of standing the sample under a
relative humidity of 93%.
[0046] FIG. 13 shows the change in X-ray diffraction pattern of the
methanesulfonate solvate (ethanol content 0.12%) of Example 5 under
various relative humidities, that is, relative humidity of 93% (1),
relative humidity of 52% (2) and relative humidity of 11% (3),
respectively.
BEST MODE FOR CARRYING OUT THE INVENTION
[0047] The present inventors have performed several experiments in
order to identify the moisture content and physicochemical property
of the methanesulfonate anhydrate and each hydrate, and the results
are described in connection with the drawings in the following.
[0048] FIG. 1 shows the moisture sorption velocity profile of
7-(3-aminomethyl-4-methoxyiminopyrrolidin-1-yl)-1-cyclopropyl-6-fluoro-4--
oxo-1,4-dihydro-1,8-naphthyridine-3-carboxylic acid
methanesulfonate anhydrate at several relative humidities. Over the
whole range of relative humidity tested, the initial moisture
adsorption proceeds rapidly at each relative humidity. In most
cases equilibrium is achieved within 2 hours. FIG. 2 shows the
isothermal moisture sorption profile of the methanesulfonate
anhydrate according to the change in relative humidity at
25.degree. C. The weight increment (%) of Y-axis represents the
equilibrium moisture content, from which it can be recognized that
the equilibrium moisture content depends on the relative humidity.
FIG. 3 shows the equilibrium moisture content of the n=3 hydrate
(which is obtained by recrystallization from a solvent mixture of
ethanol and water) after it is allowed to stand for 2 weeks under
relative humidities in the range of 23 to 75%. The result shows
that the n=3 hydrate is more stable than the anhydrate since it
maintains a moisture content of around 10% under the relative
humidities tested. FIG. 4 shows the isothermal moisture adsorption
profile of the n=1.5 hydrate. Here, it maintains a moisture content
of around 5% under the relative humidity in the range of 23 to 64%.
Thus, it is also identified as a stable hydrate.
[0049] It has been identified that the physical properties of the
hydrate are very different from those of the anhydrate.
[0050] For example, by comparing the powder X-ray diffraction
patterns of the anhydrate in FIG. 5, the n=3 hydrate in FIG. 6, and
the n=1.5 hydrate in FIG. 7, it can be seen that their crystal
forms are different from each other. In addition, the thermal
analysis using Differential Scanning Calorimetry (DSC) shows that
the endothermic peak produced by the vaporization of the water
molecules contained in the n=3 hydrate begins at around 50.degree.
C. and the exothermic peak by thermal decomposition is observed at
around 185 to 220.degree. C., whereas the anhydrate shows only an
exothermic peak at around 185 to 220.degree. C. due to the thermal
decomposition without any endothermic peak (see, FIG. 9). At the
same time, the thermogravimetric analysis shows a weight decrement
at the temperature range of endothermic peak, the extent of which
corresponds to the moisture content quantified by Karl-Fisher
method (Mettler Toledo DL37KF Coulometer)(see, FIG. 10). Therefore,
it is verified that the endothermic peak shown in the DSC analysis
is due to the evaporation of a water molecule.
[0051] The present inventors also compared the chemical stability
under heating of the hydrates with that of the anhydrate in order
to identify the influence of hydration on the chemical stability.
In this test, the anhydrate and hydrate were each kept at
70.degree. C. for 4 weeks, and the extent of decomposition is
analyzed by liquid chromatography. No difference in the extent of
decomposition was noticed between the hydrates and the anhydrate,
and thus confirming that the hydrate has the same chemical
stability as the anhydrate.
[0052] The methanesulfonate anhydrate or a solvate thereof may be
converted into a hydrate under appropriate conditions as described
above. This process can be monitored by the change in the X-ray
diffraction pattern of the compound and the decrease in the amount
of organic solvent in the compound. Such changes being caused by
the water molecules newly intercalated into the crystal
structure.
[0053] As can be seen from FIG. 11, the X-ray diffraction peaks
based on the solvate disappear with the passing of humidified
nitrogen gas to leave the peaks based on the hydrate. This shows
that all the solvates is converted into hydrates. The residual
solvent is decreased to the amount of less than the quantitative
limit simultaneously with the change of X-ray diffraction. FIG. 12
shows that the X-ray diffraction peaks based on the solvate
disappear when the solvate is allowed to stand under a relative
humidity of 93%. However, there is no change in the X-ray
diffraction pattern when the solvate is allowed to stand under a
relative humidity of 11% or 52% (see FIG. 13). Therefore, it is
recognized that the change shown in FIG. 12 occurs not by the
spontaneous evaporation of the residual solvent but by the
substitution of the organic solvents in the crystal by water
molecules.
[0054] In preparing the hydrate according to the processes
described above, the respective hydrates having a different
hydration number can be obtained by changing conditions such as
humidity, time, temperature, etc. or by changing the
recrystallization condition. Such conditions should be adjusted
according to whether the starting material is the anhydrate or a
solvate, and depending on the nature of the solvate.
[0055] The present invention will be more specifically explained by
the following examples and experimental examples. However, it
should be understood that the examples are intended to illustrate
but not in any manner limit the scope of the present invention.
Example 1
Synthesis of
7-(3-aminomethyl-4-methoxyiminopyrrolidin-1-yl)-1-cyclopropyl-6-fluoro-4--
oxo-1,4-dihydro-1,8-naphthyridine-3-carboxylic Acid
Methanesulfonate Anhydrate
[0056]
7-(3-Aminomethyl-4-methyloxyiminopyrrolidin-1-yl)-1-cyclopropyl-6-f-
luoro-4-oxo-1,4-dihydro-1,8-naphthyridine-3-carboxylic acid (3.89
g, 10 mmol) was suspended in a mixture of dichloromethane and
ethanol (110 ml, 8:2 v/v). Methanesulfonic acid (0.94 g, 9.8 mmol)
was added dropwise and the resulting solution was stirred for 1
hour at 0.degree. C. The resulting solid was filtered, washed with
ethanol then dried to give the title compound (4.55 g).
[0057] m.p.: 195.degree. C. (dec.)
[0058] .sup.1H NMR (DMSO-d.sub.6) .delta. (ppm): 8.57 (1H, s), 8.02
(1H, d), 7.98 (3H, br), 4.58 (2H, br), 4.39 (1H, m), 3.91 (3H, s),
3.85 (1H, m), 3.71 (1H, m), 3.42 (1H, m), 3.20.about.3.10 (2H, m),
1.20.about.1.10 (4H, m)
Example 2
Synthesis of
7-(3-aminomethyl-4-methoxyiminopyrrolidin-1-yl)-1-cyclopropyl-6-fluoro-4--
oxo-1,4-dihydro-1,8-naphthyridine-3-carboxylic Acid
Methanesulfonate n=3 Hydrate
[0059] A sonicator filled with water was adjusted to 40.degree. C.,
sealed with a lid and a nitrogen inlet and outlet connected. When
the pressure of the dried nitrogen introduced through the inlet was
20 psi the relative humidity of the nitrogen exiting through the
outlet was more than 93%. The anhydrate of Example 1 having a
moisture content of 2.5% (1.0 g) was introduced into a fritted
filter and the humidified nitrogen produced as described above
passed through the filter. Samples were taken after 0, 5, 10, 20,
30, and 60 minutes and the moisture content measured. From the
results shown in FIG. 8 it can be seen that a moisture content of
about 10% is maintained when the humidifying procedure is carried
out over about 30 minutes. The X-ray diffraction pattern of the
humidified sample was identical to that of the n=3 hydrate obtained
by recrystallization.
Example 3
Synthesis of
7-(3-aminomethyl-4-methoxyiminopyrrolidin-1-yl)-1-cyclopropyl-6-fluoro-4--
oxo-1,4-dihydro-1,8-naphthyridine-3-carboxylic Acid
Methanesulfonate n=1.5 Hydrate
[0060] The title compound was prepared by the following routes:
[0061] Route A
[0062] The anhydrate of Example 1 (1.0 g) was dissolved in a
mixture of water and acetone (17 ml, 10:7 v/v). The solvent was
slowly evaporated in darkness leaving the title compound as a solid
(0.8 g).
[0063] Route B
[0064] The anhydrate of Example 1 (5.0 g) was added to water (10
ml) and the mixture was heated to 45.degree. C. to aid dissolution.
Ethanol (20 ml) was added and the resulting solution stirred then
allowed to stand. The resulting solid was filtered and dried under
a flow of nitrogen to give the title compound (2.6 g).
Example 4
Synthesis of the hydrate from
7-(3-aminomethyl-4-methoxyiminopyrrolidin-1-yl)-1-cyclopropyl-6-fluoro-4--
oxo-1,4-dihydro-1,8-naphthyridine-3-carboxylic Acid
Methanesulfonate Solvate Using a Humidified Nitrogen Gas
[0065] A sonicator filled with water was adjusted to 40.degree. C.
and was sealed with a lid. Then, a nitrogen inlet and outlet were
connected to the vessel. When the pressure of the dried nitrogen
introduced through the nitrogen inlet was adjusted to about 20 psi,
the relative humidity of the humidified nitrogen gas exiting
through the outlet was more than 93%. The solvate (1 g, ethanol
0.11%) of the anhydrate of Example 1 was introduced into a fritted
filter and the humidified nitrogen gas prepared as described above
was passed through the filter. Samples were taken after 40 minutes,
3.5 and 6 hours, respectively. The change in the amount of residual
organic solvent and X-ray diffraction pattern with the lapse of
time were examined. After 3.5 hours, it was identified that the
product contained the organic solvent in an amount of less than 50
ppm and that the peaks based on the solvate disappeared, whilst the
peaks based on the mixture of n=3 hydrate and n=1.5 hydrate
appeared.
Example 5
Synthesis of the Hydrate from
7-(3-aminomethyl-4-methoxyiminopyrrolidin-1-yl)-1-cyclopropyl-6-fluoro-4--
oxo-1,4-dihydro-1,8-naphthyridine-3-carboxylic Acid
Methanesulfonate Solvate Using a High Relative Humidity
[0066] Saturated aqueous potassium nitrate solution was placed in a
desiccator, and accordingly the relative humidity inside the
desiccator was controlled to 93%. For tests under relative humidity
of 11% or 52%, desiccators containing saturated aqueous solutions
of lithium chloride and magnesium nitrate, respectively, were
prepared. Into the desiccator having a relative humidity of 93% was
introduced a solvate (1.9% ethanol) of the anhydrate of Example 1,
and into each of the desiccators having a relative humidity of 93%,
52% or 11% was introduced a solvate (0.12% ethanol) of the
anhydrate of Example 1. The solvates were stored so as not to
directly contact the aforementioned salt solutions. After a certain
period of time has passed, samples were taken and subjected to gas
chromatography in order to analyze the residual solvent. As a
result, it was identified that solvates stored for 4 weeks under a
relative humidity of 93% contained the organic solvent in an amount
of less than 50 ppm. Also, it was identified by X-ray diffraction
pattern that peaks based on the solvates disappeared after 4 weeks.
To the contrary, in the case where the samples were stored under a
relative humidity of 52% or 11%, the amount of residual organic
solvent and X-ray diffraction pattern after 4 weeks were identical
with those at the beginning.
Example 6
Synthesis of n=3 Hydrates from
7-(3-amino-methyl-4-methoxyiminopyrrolidin-1-yl)-1-cyclopropyl-6-fluoro-4-
-oxo-1,4-dihydro-1,8-naphthyridine-3-carboxylic Acid
Methanesulfonate Solvates
[0067] Dried nitrogen gas and humidified nitrogen gas having a
relative humidity of 78 to 84% were passed over 24 hours,
respectively, through 10 g of four different solvates each of which
had a different kind and amount of organic solvent from the others.
The amount of residual organic solvent was measured and the change
in X-ray diffraction pattern was analyzed, the results of which are
shown in Table 2. The X-ray diffraction analysis shows that the
samples through which dried nitrogen gas was passed remained as the
original solvates, while the samples through which humidified
nitrogen gas was passed had the same X-ray diffraction pattern and
crystallinity as those of the n=3 hydrate obtained by
recrystallization.
[0068] The results from this Example suggests that water molecules
contained in the humidified nitrogen gas replace the organic
solvents in the solvate. This suggestion is also supported by the
change in X-ray diffraction pattern influenced by a relative
humidity.
TABLE-US-00002 TABLE 2 The kind and amount of the The kind and
amount of the Sam- residual organic solvent after residual organic
solvent after ple dried nitrogen gas has passed humidified nitrogen
gas (78~84% No. for 24 hours RH) has passed for 24 hours 1
Methylene chloride 1.14%, 0.08% Ethanol 3.73% <50 ppm 2
Isopropanol 0.45% 0.06% 3 2-Isopropanol 0.24% 0.04% 4
2-Methyl-2-propanol 0.07% 0.01% Ethanol 0.06% <50 ppm
Example 7
Synthesis of the Ethanolate Containing Ethanol 0.11%
[0069] The anhydrate of Example 1 (5.0 g) was added to a solvent
mixture of ethanol (25 ml) and water (25 ml) and the mixture was
heated to 50.degree. C. to facilitate dissolution. Then, the
solution was cooled slowly to -3.degree. C. and allowed to stand at
that temperature for about 3 hours. The resulting solid was
filtered and washed with a solvent mixture of ethanol and water
(16.5 ml, ethanol:water=20:8, v/v) to give the title compound
quantitatively.
Test Example 1
Moisture Sorption of the Anhydrate of Example 1
[0070] The moisture sorption velocity and the equilibrium moisture
content of the anhydrate of Example 1 was determined by means of an
automatic moisture sorption analyzer (MB 300G Gravimetric Sorption
Analyzer). This instrument produces a specific relative humidity at
a specific temperature and continuously records the weight change
of a sample due to adsorption or desorption of moisture as measured
by a micro balance inside the instrument. The anhydrate of Example
1 (16 mg) was loaded onto the micro balance and the moisture
contained in the sample removed under a stream of dried nitrogen at
50.degree. C. A weight change of less than 5 .mu.g per 5 minutes
was the criterion for complete dryness. Thereafter, the inner
temperature was adjusted to 25.degree. C. and the sample tested at
5% intervals whilst varying the humidity from 0 to 95%. The sample
was considered to have reached equilibrium when the weight change
was less than 5 .mu.g per 5 minutes. FIG. 1 shows the moisture
adsorption velocity, that is the time required for the sample to
reach equilibrium at each relative humidity. As can be seen initial
moisture adsorption proceeded rapidly at each relative humidity
tested, in most cases equilibrium was reached within 2 hours. FIG.
2 shows the weight increment at each relative humidity, i.e. the
equilibrium moisture content. It is clear from FIG. 2 that the
equilibrium moisture content of the anhydrate is dependent on the
relative humidity.
Test Example 2
Thermal Analysis of the Anhydrate of Example 1 and n=3 Hydrate of
Example 2
[0071] For the Differential Scanning Calorimetry, METTLER TOLEDO
DSC821e and METTLER TOLEDO STARe System were used. The sample (3.7
mg) was weighed into the aluminum pan which was then press sealed
with an aluminum lid. Three tiny needle holes were made on the lid
and the sample tested by heating from normal temperature to
250.degree. C. at a rate of 10.degree. C./min. As can be seen from
FIG. 9, the endothermic peak due to the vaporization of the water
molecules contained in the n=3 hydrate begins at around 50.degree.
C. and the exothermic peak due to the thermal decomposition is
observed at around 180 to 220.degree. C. In contrast, the anhydrate
showed only an exothermic peak due to the thermal decomposition at
around 185 to 220.degree. C. without any endothermic peak.
[0072] In the thermogravimetric analysis, SEIKO TG/DTA220 was used.
The sample (3.8 mg) was weighed into an aluminum pan and was heated
from normal temperature to 250.degree. C. at a rate of 10.degree.
C./min according to the temperature raising program. As can be seen
from FIG. 10, weight decrement was observed at the temperature
range of endothermic peak, the extent of which corresponds to the
moisture content determined by Karl-Fisher method (Mettler Toledo
DL37KF Coulometer).
Test Example 3
Equilibrium Moisture Content Determination of Hydrates
[0073] Six saturated aqueous salt solutions were introduced into
each desiccator to control the inner relative humidity to a
specific value as shown in Table 3. Then, equilibrium moisture
contents of n=3 hydrate and n=1.5 hydrate of Examples 2 and 3,
respectively, were determined at several relative humidities.
TABLE-US-00003 TABLE 3 Saturated salt solutions inside the
desiccator Salt Solution Relative Humidity (%) at 25.degree. C.
Potassium Acetate 23 Magnesium Chloride 33 Potassium Carbonate 43
Magnesium Nitrate 52 Sodium Nitrite 64 Sodium Chloride 75
[0074] The sample (100 mg) was spread on a pre-weighed Petri dish
and the total weight was accurately measured, then three of the
sample were placed in each desiccator of Table 3. The desiccators
were allowed to stand at normal temperature for 7 days and then the
sample was taken to be weighed. After 13 days, one of the three
samples inside each desiccator was taken and the moisture content
of each was measured by the thermogravimetric analysis described in
Test Example 2. Equilibrium moisture content at each relative
humidity is represented in FIG. 3 (n=3 hydrate) and FIG. 4 (n=1.5
hydrate). FIG. 3 shows that moisture content of the n=3 hydrate is
maintained around 10% for the whole relative humidity range tested;
FIG. 4 shows that the moisture content of the n=1.5 hydrate is
maintained around 5% at the relative humidity of 23 to 64%. Both
hydrates are stable since they maintain a constant equilibrium
moisture content regardless of the relative humidity change.
Test Example 4
X-Ray Diffraction Analysis
[0075] The anhydrate of Example 1, n=3 hydrate of Example 2 and
n=1.5 hydrate of Example 3 (50 mg of each) were thinly spread on
the sample holder, X-ray diffraction analysis (35 kV.times.20 mA
Rigaku Gergeflex D/max-III C) were performed under the conditions
listed below. [0076] scan speed (2.theta.) 5.degree./min [0077]
sampling time: 0.03 sec [0078] scan mode: continuous [0079]
2.theta./.theta. reflection [0080] Cu-target (Ni filter)
[0081] Results of X-ray diffraction analyses on the anhydrate, n=3
hydrate, and the n=1.5 hydrate are shown in FIGS. 5, 6, and 7. The
diffraction patterns illustrate the difference in crystal form of
these 3 compounds.
[0082] According to a further aspect of the invention we provide
7-(3-aminomethyl-4-methoxyiminopyrrolidin-1-yl)-1-cyclopropyl-6-fluoro-4--
oxo-1,4-dihydro-1,8-naphthyridine-3-carboxylic acid
methanesulfonate having an X-ray diffraction pattern substantially
as shown in FIG. 5, 6 or 7.
[0083] We also provide
7-(3-aminomethyl-4-methoxyiminopyrrolidin-1-yl)-1-cyclopropyl-6-fluoro-4--
oxo-1,4-dihydro-1,8-naphthyridine-3-carboxylic acid
methanesulfonate hydrate having peaks at 2.theta.=8.0, 12.2 and
14.7.degree. in its X-ray diffraction pattern; and
7-(3-aminomethyl-4-methoxy-iminopyrrolidin-1-yl)-1-cyclopropyl-6-fluoro-4-
-oxo-1,4-dihidro-1,8-naphthyridine-3-carboxylic acid
methanesulfonate hydrate having peaks at 2.theta.=7.7 and
11.8.degree. in its X-ray diffraction pattern.
[0084] The change of crystallinity during the conversion from the
solvate to the hydrate in Examples 4 and 5 was identified by X-ray
diffraction analysis under the same conditions as mentioned above
(see, FIG. 11 to 13). FIG. 11 shows the X-ray diffraction pattern
of the solvate is changed into that of the n=3 hydrate (see,
Example 4); FIG. 12 represents the change in X-ray diffraction
pattern of the solvate containing 1.9% of ethanol before and after
storage of one week, two weeks, three weeks and four weeks at 93%
of relative humidity; and FIG. 13 represents the change in X-ray
diffraction pattern of the solvate containing 0.12% of ethanol
after storage of four weeks at 93%, 52% and 11% of relative
humidity, respectively (see, Example 5).
Test Example 5
Chemical Stability
[0085] The chemical stability of the n=3 hydrate of Example 2 and
the n=1.5 hydrate of Example 3 and the anhydrate of Example 1 were
compared at elevated temperature in order to determine the effect
on chemical stability of the extent of hydration.
[0086] The anhydrate and each of the hydrates were introduced into
a glass vial and maintained at 70.degree. C. The extent of
decomposition with elapsed time was analyzed by liquid
chromatography. The results obtained are shown in Table 4.
TABLE-US-00004 TABLE 4 Thermal stability with elapsed time (at
70.degree. C., Unit: %) Time(week) Sample Initial 1 2 3 4 Anhydrate
100 99.8 98.6 97.7 96.7 n = 3 hydrate 100 102.4 100.7 99.2 99.2 n =
1.5 hydrate 100 97.3 95.8 97.2 96.2
[0087] As can be seen from Table 4, the n-=3 hydrate and the n=1.5
hydrate both show the same degree of chemical stability as the
anhydrate.
Test Example 6
In Vitro Antibacterial Activity
[0088] In order to determine whether
7-(3-aminomethyl-4-methyloxyiminopyrrolidin-1-yl)-1-cyclopropyl-6-fluoro--
4-oxo-1,4-dihydro-1,8-naphthyridine-3-carboxylic acid
methanesulfonate has the same antibacterial activity as the free
base, in vitro antibacterial activity of the methanesulfonate was
measured using agar medium dilution method. The results are shown
in Tables 5. The minimum inhibitory concentration (MIC, .mu.g/ml)
was simply calculated in the ratio of weight without considering
the molecular weight, and ciprofloxacin was chosen as the
control.
TABLE-US-00005 TABLE 5 In vitro Antibacterial activity (Minimum
Inhibitory Concentration: MIC, .mu.g/ml) Methanesulfonic Cipro-
Test Strains acid salt floxacin Staphylococcus aureus 6538p 0.016
0.13 Staphylococcus aureus giorgio 0.016 0.13 Staphylococcus aureus
77 0.031 0.25 Staphylococcus aureus 241 4 128 Staphylococcus aureus
epidermidis 887E 0.016 0.13 Staphylococcus aureus epidermidis 178 4
128 Staphylococcus aureus faecalis 29212 0.13 0.5 Bacillus subtilis
6633 0.016 0.031 Micrococcus luteus 9431 0.13 2 Escherichia coli
10536 0.008 <0.008 Escherichia coli 3190Y 0.008 <0.008
Escherichia coli 851E 0.016 <0.008 Escherichia coli TEM3 3455E
0.25 0.5 Escherichia coli TEM5 3739E 0.13 0.13 Escherichia coli
TEM9 2639E 0.031 0.016 Pseudomonas aeruginosa 1912E 0.25 0.13
Pseudomonas aeruginosa 10145 0.5 0.5 Acinetobacter calcoaceticus
15473 0.031 0.25 Citrobacter diversus 2046E 0.031 0.016
Enterobacter cloacae 1194E 0.031 0.016 Enterobacter cloacae P99
0.016 <0.008 Klebsiella aerogenes 1976E 0.13 0.13 Klebsiella
aerogenes 1082E 0.031 0.016 Proteus vulgaris 6059 0.25 0.031
Seratia marsecence 1826E 0.13 0.063 Salmonella thypimurium 14028
0.031 0.031
Test Example 7
Water Solubility of the Anhydrate of Example 1
[0089] The water solubility of the free base and various salts of
7-(3-aminomethyl-4-methoxyiminopyrrolidin-1-yl)-1-cyclopropyl-6-fluoro-4--
o xo-1,4-dihydro-1,8-naphthyridine-3-carboxylic acid, including the
methanesulfonate of Example 1, was measured at 25.degree. C. The
results are shown in Table 6.
TABLE-US-00006 TABLE 6 Water Solubility (at 25.degree. C.)
Solubility in water Sample (mg/ml) Free form 0.007 Tartrate 6.7
Sulfurate 11.4 p-Toluenesulfonate 7.5 Methanesulfonate >30
[0090] As can be seen, the methanesulfonate shows increased water
solubility compared to that of the tartrate, the sulfurate, and the
p-toluenesulfonate and the free base.
* * * * *