CA2620142A1 - Pure and stable tiotropium bromide - Google Patents

Abstract

This invention relates to solvates of tiotropium bromide having a purity of at least 99%, process for preparing such pure solvates, and their use in pharmaceutical, formulations. This invention also provides tiotropium bromide solvates containing less than about 0.15% area by HPLC of 2,2-dithienyl glycolic acid.

Description

PURE A= STABLE TIOTROPIUM BROMIDE
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application claims the benefit of. the filing date of. United States Provisional Patent Application No. 60/836,037 filed August 7, 2006; United States Provisional Patent Application No..60/835,200 filed August. 3, 2006; United States Provisional Patent Application No. 60/835,201 filed August 3, 2006; United States Provisional Patent Application No. 60/752;672 filed December 19, 2005; United States Provisional Patent Application No. 60/754,530 filed December 27, 2005; United States Provisional Patent Application No. 60/761,437 filed January 23, 2006; United States Provisional Patent Application-No. 60/774,051 filed on February 1.5, 2006; United States Provisional Patent Application No. 60/780,310 filed March 7, 2006; United States Provisional Patent Application' No. 60/830,231 filed July 10, 2006; United States Provisional Patent Application No. 60/832,189 filed July 20, 2006; United States Provisional Patent Application No. 60/851,223 filed October 12, 2006; and United States Provisional Patent Application No_ 60/852,740 filed October 18, 2006, the disclosures of which are hereby incorporated herein by reference.
FIELD OF THE'INVENTION

[0002] The present invention is directed to pure and stable Tiotropium bromide.
BACKGROUND OF THE INVENTION

[0003] Tiotropium bromide, (1a, 2(3, 4(3, 5a, 7p) -7= [(hydroxydi-2-thienylacetyl).oxy]-9,9-dimethyl-3-oxa-9-azoniatricyclo[3..3.1.0]nonane bromide or 6p,7(3-epoxy-30-hydroxy-8-methyl-1aH,5aH-tropanium -bromide, di-2-thienyiglycolate having the following chemical structure:

Me., Me N+
O Br O
H
S O

tOH
Tiotropium bromide CisHasNO4SaBr MW: 472-.4 is- an anticholinergic drug with specificity for muscarinic receptors. As a bronchodilator it provides therapeutic benefit =in=
the =treatment of asthma or chronic obstructive pulmonary disease (COPD). This pharmaceutical ingredient is administered by inhalation, and is available commercially as SPIRIVA
HandiHal.er . =
[00041 -The preparation. and crystallization of. =Tiotropium' bromide from a mixture of acetone and methanol are disclosed in US patent no. 5,610,163.
[0005] It* is reported in "Summary Basis of Approval" of the FDA (NDA 21-395) that Tiotropium bromide is subject to- non-enzymatic hydrolysis to N-methyl scopine and 2,2-dithienyl glycolic acid, as illustrated by the following scheme.
Br . = H
N
O_ 0 O OH \ t Bre O OH N
C S f{
OXOH S ! ~ + OH TIOTROPIUM dithienyl glycolic acid N-methyl scopine BROMIDE
[0006] Degradation of the Tiotropium, bromide API in capsules which were removed from their protective package and exposed to air and humidity were reported in the'early chemistry review of NDA 21-395. The level of one.degradant was reported to be lower' than 1. 0 %- ( HPLC ) .
[0007] As a result, marketed tiotropium bromide is=packed in a moisture-resistant foil blister, and it is recommended to remove the capsule -from the package only immediately before use, as exposure to moisture in the air. can cause decomposition.
[0008] Like any synthetic compound, Tiotropium bromide can contain extraneous compounds or impurities that can -come from many sources. Some 'of these extraneous compounds or impurities may be unreacted starting materials, by-products of the reaction including the products of side. reactions, or degradation products;- vuherein the degradation products are. related to the stability -of. the API during storage.--Impurities in Tiotropium bromide or any active pharmaceutical ingredient (API) are undesirable and, in extreme: cases, might even be harmful to a patient being treated.with.a dosage form containing the API.
[0009] Therefore, there is a need- in the art for stable Tiotropium brornide, arid also for pure Tiotropium bromide solvates.
SUMMARY OF INVENTION .
[0010] In accordance with one aspect, the present invention provides a Tiotropium bromide solvate of the following.formulas:
Me Me \N,e O
Br O
H Solvent o (~tOH

having a purity of at least 99% area by HPLC.
[.0011] In another aspect, the present invention provides a Tiotropium bromide 'solvate, with less than about 0.15% area by HPLC of 2,2-dithienyl glycolic'acid. of the following formula-.

OH
O OH
s -~- D
s [0012] In- yet another aspect, the present 'inventiori provides , Tiotropium bromide solvate with a purity of at least 99% area as measured-by.HPLC, and with less than about 0.15%'area as measured by HPLC of 2,2-dithienyl glycolic acid [0013] In one aspect, the present invention provides stable Tiotropium bromide solvate.
[0014] In another aspect, the present invention provides stable Tiotropium bromide solvate with a-purity o.f at least 99%
area as measured by HPLC, and with less than about 0.15% area by HPLC of 2,2-dithienyl glycolic acid.
[001,5] In yet another aspect, the present invention provides an HPLC method for determining the purity of Tiotropium bromide solvate, and the amount of 2,2-dithienyl glycolic acid- in Tiotropium bromide. 'The method comprises: (a) combining -Tiotropium bromide sample with a mixture of acetonitrile:acetic acid in water in a ratio.of about 0.1%:99.9%, to obtain a solution;
(b)- injecting the solution into a 250X4.6 mm 'X'0.5 pm CPS
Hypersil (or similar) column;
(c) eluting the sample from the column at about 3.63 min using a mixture of perchloric acid:water in a- ratio of 7:3 (referred to as eluent A) and acetonitrile (referred to as.eluent B) as an eluent; and (d) measuring the 2.,2-dithienyl glycolic acid content in the relevant sample with a UV detector.
[0016] in one aspect, the*present.invention provid'es a process for preparing stable Tiotropi_um bromide solvate having-a purity of at least 99%.area by HPLC and with less'than aboutØ15% area of 2;2-dithienyl- glycolic acid, as measured by. HPLC, comprises crystallizing Tiotropium bromide from a solvent system comprising an organic -acid wherein the ratio of *Tiotropium bromide to the solvent system is of at least about 1 to about 5, respectively.
Organic acids which may be utilized as part of the present invention include, but are not limited to, acetic.acid,. propanoic acid, oxalic acid, maleic acid, fumaric acid, and ta.rtaric acid.
in a preferred embodiment, the organic acid.is acetic'acid.
[0017] In another aspect, the present inverition provides process for preparing stable Tiotropiurn bromide solvate having a purity of at least 99R; area by HPLC and with .less than about 0.05% area of 2,2-dithienyl glycolic acid as measured by HPLC, comprises crystallizing Tiotropium bromide from a sol=vent. system comprising an organic acid wherein the ratio of Tiotropium bromide to the solvent system is of at least about 1 to about 10, respectively: Organic acids which may be utilized as part of the present invention include, but are not limi-ted to,=acetic acid, propanoic acid, oxalic.acid, maleic acid, fumaric acid, and-tartaric acid. in a pref,erred = embodiment,* the organic acid is.
acetic acid.
[0018] In yet another'aspect, the laresent invention=provides a~
process for preparing stable Tiotropium bromide solvate having a purity of at least. 99% area by HPLC and "with less' than about 0.02% area of 2,2-dithienyl glycolic acid as measured by HPLC,.
comprises crystallizing Tiotropium bromide from a solvent system comprising an organi.c= acid wherein the ratio= of Tiotropium bromide to 'the solvent system is of at least about 1'to about 20, respectively. Organic acids which may be utilized as part of the present invention include, but are not limited to-,. acetic 'acid, propanoic acid, oxalic acid, maleic acid, fumaric acid, and tartaric. acid. -in a preferred embodiment-, the organic acid is ' acetic acid. .[0019] In one aspect, the present invention provides a process for preparing stable Tiotropium -bromide solvate having a purity of at least 99% area by HPLC and with less than about -0.01% area of 2,2-dithienyl glycolic acid as, measured by HPLC, comprises crystallizing Tiotropium bromide from a solvent system.comprising an organic acid wherein the ratio of Tiotropium bromide to the solvent sys.tem ' is of at least about 1 to. about 30, respectively.
Organic acids which 'may be utilized as -part of the present invention include, but are not limited to, acetic acid,.*propanoic acid, oxalic acid, maleic acid, fumaric acicl, and ta=rtaric acid.
in a preferred embodiment, the organic acid is acetic acid;
[0020] In another aspect, the present invention provides a.
pharmaceutical, composition comprising stable Tiotropium bromide solvate with a purity of at least 99% area as measured by HPLC, and with less than about 0.15% area as measured by HPLC of 2,2-dithienyl glycolic acid, and pharmaceutically acceptable excipients.' .[0021] In yet-another aspect, the present invent-ion provides a proeess for preparing pharmaceutical composition comprising stable Tiotropium bromide solvate with a purity of at least 99%' area as measured by HPLC, and with less than about 0.15% area as measured by - HPLC -of 2,2-dithienyl glycolic acid, and pharmaceutically acceptable excipients.
[0022] In one aspect, the present invention provides the use of the stable Tiotropium bromide solvate with a -purity"of at least 99% area as measured by. HPLC, and with less than about 0.15% area by HPLC of 2,2-dithienyl glycolic acid,. of the present invention for the.manufacture of a pharmaceutical.composition., [0023] In yet another embodiment, the present ..invention encompasses a process for preparing Tiotropium bromide with less than about Ø.15% area by -HPLC of 2,2=dithienyl glycolic acid' comprising the steps of -(a) ~obtaining one or more samples of one or more Tiotrpium bromide batches;
(b) measuring the level of 2,2-dithienyl glycolic acid in each of the samples of (a);
.(c). selecting the.Tiotropium bromide batch that comprises-a level of 2,2-dithienyl glycolic acid of less than about 0.-15% area by HPLC, based on the measurement or measurements conducted.in step (b); and (d) using. the batch selected in step (c) to prepare said any Tiotropium bromide comprising less than =about 0.15%
area by HPLC of 2',2-dithienyl glycolic acid. .

DETAILED DESCRIPTION OF THE INVENTION
[0024] An analysis of Spiriva11H. capsules:; containing monohydrate tiotropium.bromide,-supports the hydrolysis theory of Tiotropium bromide degradation by demonstrating-that the level of dithienyl glycolic acid in Tiotropium bromide monohydrate is above 0.7% area by HPLC.* in contrast, the present invention .succeeds to provide substantially pure. Tiotropium bromide solvate. The present invention also provides stable Tiotropium bromide solvate i.e., Tiotropium bromide solvate that is significantly less prone to"hydrolysis upon storage. According to general ICH guidelines, acceptable purities of drug products for human treatment are generally greater than-99.0% area by HPLC, and preferably greater than 99.55% area as measured by HPLC, where the conterit of each single impurity is preferably less than about 0.15% area as measured by HPLC. Thus, providing such pure Tiotropium bromide solvate, which is also less prone to decomposition. upon storage, is preferable considering- that prolonged shelf life-is an advantage for industrial manufacturing and safety of patients.
[0025] The present invention provides Tiotropium bromide solvate of the following formula:

Me ~ Me Ni!

Br O

O
H Solvent OH
s having a purity -of at least 99% area by HPLC.= PreferabTy,' the above Tiotropium bromide has purity ranging from about 99% area to about 100% area as measured by HPLC,-.more preferably ranging from about 99.5% area'to about 100% area as-measured by HPLC, and most preferably ranging. from. about 99 . 7% area -to -about.'100% =area as measured by HPLC. One skilled in the art'would recognize that the solvate:may contain any number of solvent molecules. , [0026] . Typically, the.term "solvate" as used herein,*refers to a substance that includes any solvent other than water at levels of more than 1%. 'Preferably, the solvate form of Tiotropium bromide is selected from a group consisting of an alcoholate and an acetic acid solvate. Preferably, the alcoholate is a C1_$' alcoholate, more preferably a* C1_6 alcoholate, even . more preferably a Cl_5 alcoholate, -and most preferably a C1_4 alcohoTate:
Preferably, the C1_4 alcoholate is selected . from the. group consisting of methanolate, ethanolate, isopropanolate, n-propanolate and n-butanolate. Most preferably, the Cl_4 alcoholate is methanolate,- ethanolate or n-propanolate.
[0027] -The present invention also provides Tiotropium bromide*
solvate containing less than about 0.15% area of 2,2 -dithienyl' glycolic acid as measured by HPLC. Preferably, the' said Tiotropium bromide is with about 0.15% area by HPLC to -the=
detection limit of an HPLC method of 2=,2-di=thienyl glycolic acid.
[0026] The terms "detection limit" or "detection limit of an HPLC method" refer to any. HPLC method used to determine the purity of Tiotropium bromide, and in particular, to determine the amount of 2,2-dithienylglycolic acid in any Tiotropium bromide sample. Preferably, the detection limit is the detection limit of the HPLC
method used in the present invention-, or of any other equivalent method. [0029] Preferably, Tiotropium bromide solvate is containing less than about 0.05% area as measured by'HPLC of 2,2-dithi.enyl, glycolicacid, more preferably, the said Tiotropium bromide is with about -0.05% area by HPLC to the detection limit of *an HPLC
method of 2,2-dithienyl glycolic acid.
[0030] Preferably, Tiotropium bromide solvate is containing less than about 0.02% area as measured'by HPLC of 2;2=-dithienyl glycolic acid, more preferably, the said Tiotropium bromide is with about 0.02% area by HPLC to the detection limit of an HPLC
method of 2,2-di=thienyl glycolic acid.
[00311 Preferably, Tiotropium bromide solvate. is containing less than about 0.01% area as measured by HPLC of 2,2-dithienyl glycolic . acid, more preferably, the said Tiotropium=bromide is with about 0. 01 % area by HPLC to the detection limit = of an HPLC
method, of 2,2-dithienyl glycolic acid.
[00321 The present invention further provides Tiotropi-um bromide solvate with.a purity of at least 99% area as measured by ' HPLC, and containing less than about 0.15% area as measured :by HPLC of 2,2-dithienyl glycolic acid.
E00331 The present: invention also provides* stable Tiotropium bromide solvate. As used herein, the term "stable;." in reference to Tiotropium bromide, means Tiotropium bromide wherein the level of a specific impurity does not'increase to more than a specific limit, when maintained at a specific relative humidity and temperature for a. specific period of time. More specifically,.
the term "stable" means Tiotropium bromide wherein .the-level of-the 2,2-dithienyl glycolic acid, shown below, does not increase' ==to more than 0.15~ of the total amount'of tiotropium bromide area as measured by HPLC, when maintained at a temperature ranging.
from about 42C to about 302C, for at least about two months.
OH
O OH
S
S

[0034] The present invention provides stable Tiotropium bromide solvate with a purity of at least 99% area as measured by HPLC,and containing less than about' 0.15% area as measured by HPLC of 2,2-dithienyl glycolic acid.
[0035]The stability and.-purity of Tiotrop'ium bromide vaere tested. Data -indicated that when storing Tiotropium bromide monohydrate-at a temperature above 44C, an' increase in' the content of the 2,2-dithienyl glycolic acid ' was-- detected.
However, - 'when Tiotropium bromide solvate, such as hemi-ethanolate, was stored at a temperature above 49C,. the presence of 2,2-dithienyl glycolic acid was detected only ..after -two months, and even then, the level was significantl-y lower than.the level detected in the monohydrate product.
[0036] The purity of Tiotropium bromide, as well as the amount of- 2,2-dithienyl glycolic acid in Tiotropium' bromide.' is-determined by an-HPLC method comprising: (a) . combining. 'Tiotropium bromide -sample with a mixture of acetonitrile:acetic acid in water in a ratio of about 0.1%:99.9%, to obtain a solution;

(b) injecting the solution into -a 250X4.6 mm X0.5 p.m CPS
Hypersil (or similar) column; (c) eluting the'sample from the column at about 3.63 min using an eluent mixture of perchioric acid:water in, a ratio of 7:3 (referred to as eluent A) and acetonitrile (referred to as eluent B) as an eluent;. and ' -(d) measuring the 2,2-dithienyl- glycolic-acid content in .the relevant sample with'a W detector.
[0037] The eluent used may be a mixture of eluent A and eluent B, =wherein .theratio of them varies over the time, '. i.e. a gradient eluent. At the time 0 minutes, the eluent contains 70%
of eluent A and 300% of eluent B. At 23 miriutes, the ' eluent contains 55% of eluent A and 45% of eluent B. At 30.minutes, the eluent contains 50% of eluent A and-50% of eluent B. At 35 minutes, the'eluent contains-50% of eluent-A and_50% of eluent B.
At 40 -minutes, the eluent contains .35% of eluent A. and ' 65% of eluent B,.and.at 41 minutes, the eluent contains 70%,of eluent A
and 30% of eluent B. . .
[0038] Preferably, the 2,2-dithienyl glycolic acid content-is measured at a wave length of 240 nm.
[0039] Such pure and stable Tiotropium bromide solvates can be prepared by a process comprises crystallizing Tiotropium bromide from a suitable solvent system; whereiri the s'olvent system comprises. acetic acid. .
[0040] The process for_ preparing stable T-iotropium bromide solvate,having a purity of at least 99% area by HPLC and with less than about 0.15% area of 2,2-dithienyl glycol=ic acid as measured by HPLC, comprises crystallizing Tiotropium bromide from a solvent system comprising acetic acid; wherein the ratio of Tiotropiunu bromide to the solvent system is of at least about 1 to about 5, respectively. [0041]. The process for preparing stable Tiotropium=
bromide solvate having a purity of at least 99% area by HPLC and with less than about 0.05% area of 2,2-dithienyl glycolic acid as measured by HPLC, comprises crystallizing Tiotrop.ium bromide from a solvent system comprising an=organic acid wherein the ratio of Tiotropium bromide to the solvent system is of at. least about. 1- .
to about 10, respectively. Organic acids which.may be utilized as part of the 'present.invention include, but are not. limited to, acetic acid, = propanoic =.acid, oxalic ''acid, maleic acid, fumaric acid, and _ tartaric acid. In a preferred embodiment, the.organic-acid is acetic acid. [0042] The process for preparing stable Tiotropium bromide solvate having a purity of at least 99% area by HPLC and with less than about.. 0.02%= area of 2,2-dithienyl glycolic acid as measured by HPLC, comprises crystallizing Tiotropium bromide from a solvent.system comprising an organic acid-wherein the ratio of Tiotropium bromide to the solvent system is of at'least about 1 to about 20, respectively: Organic acids which may be utilized as part of the present invention include, but are not limited to, acetic acid, propanoic acid, oxalic =acid, maleic. 'acid; fuinaric acid, and tartaric acid. In a preferred embodiment, the organic acid is acetic acid:
[00437 The process for preparing stable Tiotropium bromide solvate having a purity of at least 99% area by. HP.LC.'-and with, less than -about 0.01% area of 2,2-dithienyl glycolic= ac=id as measured-by:HPLC, comprises crystallizing Tiotropium..bromide,from a-solvent system comprising an organic acid wherein the ratio of Tiotropium bromide to the solvent system. is of at le'ast about 1 to about 30, respective.ly. Organic acids which may be utilized as part of the present invention include, but' are not limited. to, acetic acid, propanoic acid, oxalic acid, maleic acid, fumaric acid, and tartaric acid.' In a preferred embodiment,-the -organic -acid. is..
acetic acid. -[00443 Preferably, the crystallization process comprises providing a solution of tiotropium bromide in the so].vent system' comprising organic acid, and cooling to obtain asuspension.
Preferably,' the organic.acid is acetic.acid.
[00453 -In yet another embodiment, the present invention"
encompasses a pr-ocess-for preparing Tiotropium bromide.with less-than about 0.15% area by HPLC of 2,2-dithienyl glycolic acid comprising the steps of . = .
(a) obtaining one or more samples of one or more'Tiotrpium bromide batches.;
(b) measuring the level of 2,2-dithienyl= glycolic,acid in each of the samples of (a);
.(c) selecting the Tiotropium bromide batch that comprises*a level of 2,2-dithienyl glycolic acid of less- than about 0..1.5%. area by HPLC, based on the measurement. or measurements=conducted in step (b);and (d) using the batch -selected instep (c) to prepare said' any Tiotropium bromide comprising. less than about 0.15%
area by HPLC of 2,2-dithienyl glycolic acid:

[0046] Typically, the Tiotrpium bromide of step '(a) comprises a sufficiently low level of 2,2-dithienyl glycolic: acid. More' preferably, the. Tiotrpium bromide of step (a) comprises _less than about 0.15% area by HPLC of 2,2-dithienyl glycolic acid-.
[0047] When the sample of Tiotrpium bromide of formula II of step (a) comprises' more than about 0.15% area -by HPLC of 2,2=
dithienyl glycolic acid, according to the measurement in step (b), the sample may be purified, prior to performing step (c).*
[0048] Typically, the purifed Tiotropium bromide.=comprises a lower level of 2,2-d'ithienyl glycolic acid than the level present before purification. Preferably, the tiotrpium bromide sample of step (a) obtained after purification,- comprises less than =about=
0.15% areaby HPLC of 2,2-dithienyl glycolic=acid.=
[0049] i.7nless specified otherwise, the tiotropium bromide of step (d) of the above process may be any form tiotropium bromide, including, for example, crystalline forms and amorphous form of -tiotropium bromide.
[0050] The Tiotropium bromide used as.a sta.rting material for the crystallization processes may be prepared, for =example', according to -tZie process disclosed in Co-pending application No. 60/835,201 entitled 'PROCESS FOR THE PREP.ARATION- OF' TRIOTROPIUM BROMIDE filed in the U.S. Patent and Trademark Office on August 3, 2006, according to the proces=s disc=losed in Co-pending application No. 60/830,231 entitled PROCESS FOR THE
PREPARATION -OF TIOTROPIUM BROMIDE filed in the 'U.S. Patent and=
Trademark Office oin Jizly. 10, '2006, or by any other process known to one skilled in the art. [0051] =- The. process disclosed in Co-pending application Nos. 60/830,231 -and 60=/835,201 disclose combining- methyl-di-(2-thienyl)-glycolate of formula I, an inorganic base, a polar organic solvent, and scopine salt'of formula II-s, xe =
O OMe NH
OH H. ~ =
s C S H

I .= =II--s containing aboutØ5% to about 40% of salts;'heating; recovering;
and adding'an organic solveiit. [0052] The process may be performed according to the following s cheme : . . = Xe NH
H Br e ~ N iN .
0 OMe OH ' H _ H
SCOPINE salts II-s 0;~ O
OHS O O MeBr _ .. O O
~ S KZC03, DMF ~\OH Acetnnitrile OH
t_S \ S ~ ~
METHYL DI(2-THIENY.L) S S
GLYCOLATE (I) TIOTROPIUM BROMIDE
N-DEMETHYL TIOTROPIUM (IV) (III) [00531 The glycolate of formuZa=T may be=prepared by combining.
2-bromo-thiophene of the,following formula;

Br ~ =
. .5.. , . .
Mg, and an ethereal solvent; combining with dimethyloxalate of' the following formula, '=' O ==

OMe MeO O . ly and quenching.
[0054] Combining 2-bromo-tiophene, Mg, and an ethereal,solvent provides a Grignard reagent that can be 'prepared, for example, "according to the process disclosed in Nyberg, K. Acta Chemica Scandinavica, 24,.1970,:1590-1596.
[0055] Methyl' di-(2-thienyl)glycolate of formula I may be purified' by*.crystallization from a= mixture of ethanol and heptane, absolute -ethanol and heptane, isopropanol- and. heptane, or from toluene and heptane.
[0055]* Preferably, the scopine salt of formula II-s is suspended in -a polar organic solvent. Preferably, the polar organic solvent is selected from a groiup consisting= o.f dimethylformamide, N-methyl-2-pyrrolidone,' dimethylacetamide., dimethylsulfoxide, acetonitrile, sulfolane, and mixtures thereof..
More preferably, the polar organic solvent is dimethylformamide.
Preferably, the salt is an HBr salt.
[0057] Preferably, the i.norganic base and inethyl-di-(2-thienyl)-glycolate of formula I are added to the suspension. More preferably, the inorganic base is -anhydrous. Even more preferably, the inorganic base has a pKa of about 6 to about 12, more preferably of about 9 to about 10. Yet even more preferably, the inorganic base is selected from a group consisting -of : K2C03;
NaHCO3, Na2CO3, Li2CO3, Cs2CO3, KOtBu, and LiOtBu. Most preferably, the inorganic base is K2C03. The inorganic base is added in an amount of 0.45 to 2.5 mole equivalent per mole equivalent, more preferably, 2 to 2_5 mole equivalent per mole equivalent of' scopine salt.
[0058] Preferably, methyl-di-(2-thienyl)-glycolate of fvrmula I is added in the form of a solution in the polar organic solvent. Preferably, the inorganic base and methyl-di-(2-thienyl)-glycolate of formula I are added at a temperature of about 25 C to about 65 C, more preferably.at about 604C to about 654C. .
[0059] Preferably, the suspension containing all the above substances is heated to a temperature of below 702C, more preferably to a temperature ranging from about 252C to- about 65sC, even more preferably at a temperatu:re ranging from about 602C to about 652C, and most preferably at a temperature rangirig from about 632C to about 65sC. Preferably, heating is.done under reduced pressure-. Preferably, the pressure is of about 70 to .
about 100 milibar. Preferably, nitrogen is bubbled during the reaction, through a second inlet. More preferably; nitrogen is bubbled in a rate of about 1.8 to about 2.6 L/min, even more preferably of about 2.0 to about 2.4 L/min, and yet even more preferably of. about' 2.2 to about 2.4 L/min. Heating under pressure, while bubbling nitrogen from a second inlet,_ assist-s in*.
evaporating methanol, which is formed. during the reaction. As such, the reaction shifts towards the formation of.the product.
Preferably, heating is done for a time ranging from about 17 - to about 24 hours, more preferably for about.18 to about 20 hours.[0060] N-demethyi-tiotropi.um 'of formula III may 'be recovered by a) cooling the suspension; b) adding an acid; c) extracting the aqueous phase; d) adding a base to the aqueous phase; e) filtering; and f) washing and drying. Preferably, the acid is HBr. Preferably, 'the suspension is cooled to a temperature 'of about 102C to about -104C, more preferably to about 59C to about 02C. Preferably, the addition of the acid provides a pH of about 3. Preferably, the aqueous phase is extracted with toluene.
Preferably, the base is added at a temperature of about O4C to about 52C. More preferably, the base is K2C03. Preferably, the*
addition of the base causes precipitation of- N-demethyl-tiotropium of formula III. Preferably, the precipitate is washed with water to obtain a pH of about 7.
[0061]'' Optionally, scopine base may be used. When scopine base is used, preferably a smaller amount of the inorganicbase is used. Preferably; about .1 to 1.5 mole e.quivalent- of -inorganic base per mole equivalent of scopine base may beused.
[0062] After N-demethyl-tiotropium of formula III is obtained, it is converted to Tiotropium. bromide by reacting with methylbromide in an organic solvent. Preferably, the organic solvent is selected- from a group consisting of : C2_4.nitrile, C4_8 linear or cyclic ether, mixtures of C2_4 nitrile and C4_8 linear or cyclic ether, mixtures of C7_e 'aromatic hydrocarbon and C2_4 nitrile, and mixtures of C2-4 nitrile and C3_1Q ketone. Preferably, the C2-4 nitrile is acetonitrile. A preferred C4_8 linear or cyclic ether is tetrahydrofuran. Preferably, a mixture of- C2_4 nitrile and C4_8 linear or cyclic ether is that of acetonitrile- and tetrahydrofuran. A preferred mixture of C7_8 'aromatic hydrocarbon and C2_4 nitrile is that of toluene and -acetonitrile. Preferably, a mixture of C2_4 nitrile and C3-10 ketone is. that of-, acetone and acetonitrile, and heating is conducted to a temperatiire=of about 202C to about 40gC. Preferably, the=solvent is. acetonitrile.
Preferably, heating is done to a temperature of, about 202C to .about 252C.. .More, preferably, heating is done for about= 12 to about 64 hours, even more preferably, for about 18 to about 22 hours.
10063] Initially, crude Tiotropium bromide is dissolved in the solvent system which is comprised of an organic acid.Examples of such organic acids include, but are not limited to, trifluoroacetic acid, tartaric acid, maleic acid, propionic acid, oxalic acid, p-toluen sulphonic acid, methan sulphonic acid,= HCl,=
HBr, H2SO4 and acetic acid.. Preferably the organic- acid is acetic acid.
[0064] Preferably, the solvent system=comprises= acetic acid, C1_$ alcohol and acetic acid, C1_8 alcohol, acetic acid and acetone or Cl_8 alcohol, acetic acid and water. Preferably;'. ==the.
alcoholate is a C1_8 alcohol, more preferably a C1_6 alcohol, even more preferably a C1_5 alcohol, and most preferably a. Cl_4 alcohol.
Preferably, the C1_4 alcohol is selected from the group consisting of methanolate, ethanolate, isopropanolate, n-propanolate and n-butanolate. Most preferably, the C1_4 alcoholate is methanolate, ethanolate or n-propanolate.. =
[0065] Typically, the dissolution is achieved by'heating' the combination of Tiotropium bromide and -the solvent system.
Preferably, the heating is to a temperature ranging from about 604C to about 784C, more preferably from to about 654C to about 782C, most preferably from about 65QC to about 752C.
[0066] Preferably, the solution is then cooled to a temperature ranging from about 252C to about 04C; more preferably from about 254C to' about 52C, and most''preferably -from about 52C
to about 02C, to induce precipitation-'of the crystallized product . Preferably, cooling is done over a period: '.of a]aouit 4 to about 10 hours, more 'preferably from about 6, to about 9 hours, most preferably of about 8 to about 9 hours..[0067] Typically, the suspension is maintained-.to increase the yield of, the precipitated crystallized product. Preferably, the suspension is maintai-ned'for a time period ranging from at least about 3 hours to about 21 hours, more preferably from about 6 hours to about 12 hours, and.most preferably from about 13 hours to about 18 hours.
[0068] The crystallization process may further comprise a recovery step. The precipitate-may be recove'red by 'any method known to a skill"ed artisan. Preferably, the recover compris'es filtering the suspension, washing the filtered p.roduct; and drying. .
[0069] The present invention also provides a pharanaceutical composition comprising a stable Tiotropium bromide solvate having a purity of at least 99% area as- measured by HPLC, aild with less than about 0.15% area by HPLC of 2,2-dithienyl glycolic acid, and pharmaceutically acceptable excipients. The present invention also provides a. pharmaceutical.. composition comprisirig a~stable Tiotropium bromide solvate having.a purity of at least 99.3%'area as measured by HPLC, and with less than about 0.15% area.by .HPLC
of 2,2-dithienyl'glycolic acid, and pharmaceutically acceptable excipients. The-present invention.also provides a pharmaceutical compositi.on comprising a.stable Tiotropium bromide solvate having a purity of at least 99.5% area as measured by HPLC, and with~~
less than about 0.15% area by HPLC of 2,2-dithi:enyl glycolic acid, and pharmaceutically acceptable excipi.ents. ' [0070] The present invention further provides a process'for' preparing pharmaceutical composition comprising stable.Tiotropium'.
brom'ide solvate with a purity of at least 99% -area by HPLC,*and containing less than-about 0.15% area as measured by HPLC-of 2,2-dithienyl, =glycolic acid, and pharmaceutically. acceptable excipients. The pure and stable Tiotropium bromide can be' micronized to. prepare material suitable for formulation.
Typically, the term "suitable for formulation" in reterence'to micronized Tiotropium bromide corresponds to Tiotropium bromide having at least 90% of the particles below 20 microns. The micronization process can, optionally,. be followed' by a process .comprising exposing the micronized form-to a suitable solvent to restore the. initial content of solvent in the.solvate: Usually, the term "suitable solvent" corresponds to the kind of solvent in the original solvated form.*
[0071] The present invention further provides a method of treating asthma or chromic pulmonary disease by.administration of an effective- amount of a pharmaceutical composition comprising s'table Tiotropium bromide solvate having a purity of at least 99%
area as measured by HPLC,- and containing less than about 0.15%~
area as measured by HPLC, of 2,2-dithienyl glycolic 'acid, and pharmaceutical excipients..
[0072] The present invention provides the use of the stable Tiotropium bromide solvate with a purity of at least 99% area as measured by HPLC, and.containing less than about 0:15% area as measured,by HPLC..of 2,2-dithienyl glycolic acid, of the present invention.for the manufacture of a pharmaceutical composition.
[0073] . Methods of - administration- of a pharmaceutical composition of the present invention cari be administered in various preparations depending on the age,.sex, and symptoms of the patient. The pharmaceutical compositions can -be administered, for example, as tablets, pills, powders, liquids, suspensions, emulsions, granules, capsules, . suppositories, injection preparations (solutions and suspensions), and the like.
[0074] Pharmaceutical compositions of the present invention can optionally be mixed with other forms of =Tioti-opium bromide solvate and/or other active ingredients such-as HMG-CoA reductase inhibitors. In addition, pharmaceutical compositions of ..the present invention. can contain .inactive ingredients.' such as-diluents, carriers, fillers, bulking agents, binders, disintegrants, disintegration inhibitors, absorption accelerators, -wetting agents, lubricants-, glidants, surf'ace active agents, flavoring agents, and the like.' [0075] Diluents increase the bulk of a solid pharmacei.itical composition:and can make a pharmaceutical dosage form containing the composition easier for the patierit and care giveac to handle.
Diluents for, solid compositions include, for -example,.
microcrystalline cellulose (e.g., Avicel ), microfine cellulose, lactose, starch, pregelitinized starch, calcium carbonate, calcium sulfate, sugar, dextrates, dextrin, dextrose, dibasic calcium phosphate dihydrate, tribasic calcium phosphate, kaolin, magnesium carbonate, magnesium oxide, maltodextrin., mannitol, polyinethacrylates (e.g., Eudragit ), potassium chloride, powdered cellulose, sodium chloride, sorbitol, or talc.
[0076] Carriers for use in the pharmaceutical comp'ositions'may include,* but are not limited to, lactose, white sugar, sodium .chloride, glucose, urea, starch, calcium carbonate, kaolin,' crystalline cellulose,-or silicic acid. .-[0077] Binders help bind the active ingredient and other excipients together after compression. Binders for solid' pharmaceutical- compositions include for example acacia, alginic acid, carbomer (e.g. carbopol), carboxymethylcellulose, sodium, dextrin, ethyl -cellulose, gelati:n, guar gum,- hydrogenated vegetable oil, hydroxyethyl cellulose, hydroxypropyl cellulose (e.g. Klucel ), hydroxypropyl methyl cellulose (e.g. Methocel ), liquid glucose, magnesium. aluminum silicate, maltodextrin, methylcellulose, polymethacrylates, povidone. -(e.g. Koll.idon~, Plasdone ); pregelatinized starch, sodium alginate, or s-tarch.
[0078] Disintegrants can increase dissolution. Disintegrants include, for example, alginic acid, carboxymethylcellulose calcium, carboxymethylcellulose.= sodium (e.g. Ac-Di-So1 , Primellose ), colloidal silicon dioxide, croscarmellose sodium-,-crospovidone (e.g. Kollidon , Polyplasdone ), guar gum, magnesium aluminum silicate, methyl cellulose, microcrystalline cellulose, polacrilin potassium, powdered cellulose, pregelatinized starch, sodium alginate, sodium starchglycolate (e.g.. Explotab ) and starch.
[0079] Disintegration inhibitors may include, but are not limited to, -whi.te sugar, stearin, cocoin.ut' butter;- hydrogenated oils, and the like.Absorption accelerators may include, but are not limited to, quaternary ammonium base, sodium laurylsulfate, and the like. [0080] Wetting- agents may include-, but are -not limited to, glycerin, starch, and 'the like. Adsorbing- agents used include, but are not limited to, starch, lactose, kaolin, bentonite, colloidal silicic acid, and the like-. [0081] A lubricant can be added to the composition to reduce adhesion and ease release of the product from a punch or dye -during tableting. Lubri,cants include for example magnesium stearate, calcium stearate, glyceryl -monostearate, glyceryl palmitostearate,* hydrogenated castor oil, hydrogenated vegetable oil, mineral oil, polyethylene glycol, sodium benzoate, sodium lauryl sulfate, sodium stearyl fumarate, stearic acid, talc and zinc stearate. . -' [0082] Glidants can be added to improve the flowability of non-compacted solid composition and improve the accuracy of-dosing. Excipients that can func-tion as glidants -include *for example colloidal silicon- dioxide, magnesium trisilicate, powdered cellulose, starch, talc and tribasic calcium'phosphate.
.[0083] Flavoring agents and flavor enhancers make the dosage form more palatable to-the patient. Common flavoring agents and flavor enhancers for pharmaceutical products that can be included in the composition of the present-invention include' for example maltol, vanillin, ethyl vanillin, menthol, ci:tric, acid, fumaric.-acid, ethyl maltol, and tartaric acid.

[0084] Tablets can be further coated with commonly known coating materials such as sugar coated=tablets,. gelatin film coated tablets, tablets coated with enteric coatings; tablets coated with f-ilms, double layered tablets,. and multi-layered .tablets. Capsules can be coated with shell made, for' example, from gelatin -and optionally contain. a plasticizer such as glycerin-and sorbitol, and an opacifying agent or,colorant.
[0085] Solid and liquid compositions can also.be dyed using any pharmaceutically acceptable colorant to improve their appearance and/or 'facilitate 'patient identification of the product and unit dosage level.
[0086] In liquid pharmaceutical- compositions of 'the present invention, the Tiotropium bromide solvate forms -described herein and any other solid ingredients are dissolved or suspended in* a liquid carrier, such as water, vegetable oil, - alcohol, polyethylene glycol, propylene glycol or glycerin: [0087] Liquid pharmaceutical. compositions -=can contain emulsifying agents to disperse uniformly throughout the composition an active ingr.edient= or other excipient'that is not*
soluble in the liquid carrier.- Emulsifyi.ng agents that can be useful in liquid compositions of the present invention iriclude, for example,' gelatin, egg yolk, casein, cholesterol, acacia, tragacarith, chondrus, pectin, methyl' cellulose, carbomer, cetostearyl alcohol and cetyl alcohol.
Liquid pharmaceutical.compositions of the present invention can also contain viscosity enhancing agents to improve the mouth-feel of the product and/or coat the lining of 'the gastrointestinal tract. Such agents include for example acacia, alginic.acid bentonite, carbomer, carboxymethylcellulose calcium or sodium, cetostearyl alcohol, methyl -cellulose, ethylcellulose, "gelatinguar gum, hydroxyethyl cellulose; hydroxypropyl cellulose, hydroxypropyl methyl cellulose, maltodextrin, polyviny.l alcohol, povi.done, propylene carbonate, propylene glycol alginate,-sodium alginate,_sodium starch*glycolate, starch-tragacanth and'xanthan gum.
[0088] Sweetening agents such as sorbitol, saccharin, sodium saccharin, sucrose, aspartame, fructose, manni.tol 'and invert sugar can be added to improve the taste.
[0089] Preservatives and chelating agents such as alcohol, sodium benzoate, butylated. hydroxy toluene, butylated hydroxyanisole and ethylenediami.ne tetraacetic acid can be added at safe levels to improve storage stability.
[0090] A liquid composition according to the present invention can also contain a buffer such as gluconic acid, lactic acid, citric acid or acetic acid, sodium.gluconate, sodium lactate, sodium citrate.or sodium acetate. .
[0091] Selection of excipients and the amounts to use can be readily determined -by =an experienced formulation scientist in view of standard procedures and reference works known'.in the art.' [0092] A composition for tableting or capsule= filing can- be prepared'by wet granulation. In wet granulation some or all of the active ingredients and excipients in powder form are blended and then further mixed in. the 'presence of a liquid, typically water, which causes the powders to clump up into granules. The granulate. is screened and/or milled, dri-ed and then screened and/or milled to the desired particle size. The granulate can then be tableted or, other excipients can be added prior to tableting, such as a glidant and/or a lubricant.
[0093] A tableting composition can be prepared conventionally by dry blending. For instance, the blended composition .of'.the actives and excipients= can be compacted into a slug or a sheet and then comminuted into compacted granules. The compacted=
granules can be compressed subsequently into a tablet.
[0094] As an alternative to . dry granulation,=..a blended composition can be compressed - -directly into a compacted dosage form using direct compression techniques_ Direct compression produces a more uniform tablet without granules.- Excipients that are particularly well-suited to direct compression tableting include microcrystalline cellulose, spray dried lactose, dicalcium' phosphate dihydrate and colloidal silica. The proper use of these and other excipients in direct compressiori tabletincg is. known to those in the art with experience and skill in particular formulation challenges of direct 'compression tableting. [0095] A capsule filling.of the present invent-ion can comprise any of the aforementioned blends and granulates.=that were described with reference to tableting, only they are not subjected to a final tableting step."
[0096] wh.en shaping' the pharmaceutical composition into pill form, any commonly known excipient used in the.art can be.used.
For example,- carriers include, but are-not' limited to, lactose, atarch, coconut butter, hardened vegetable oils,- kaolin, talc, and- the like. Binders used .include, but are not limited to, guni arabic. powder, tragacanth gum powder, gelatin, ethanol, and the like. Disintegrating agents used include, but are not limited to, agar, lazninalia, and the like.
[0097] For the purpose of shaping the pharmaceutical composition in the form of suppositories, any cominonly known excipient-used in.the art can be used. For example,, excipients.
include, but* are not limited to, polyethylene glycols; coconut butter, higher alcohol.s, esters of higher alcohols, gelatin, .
semisynthesized.glycerides, and the like.
[0098] when preparing injectable pharmaceutical compositions,-solutions and suspensions are sterilized and are preferably.made isotonic to blood. Injection preparations may use carriers commonly known in the art. For example, carriers for injectable preparations include, but. are not limitedto, water, ethyl alcohol, propylene glycol, ethoxylated isostearyl- alcohol, polyoxylated isostearyl alcohol, and fatty acid esters of polyoxyethylene sorbitan. One of ordinary skill in the art can easily determine with little or no experimentation the amount of sodium chloride, glucose, or glycer-in necessary to make' the injectable preparation isotonic. Additional ingredients, such as dissolving agents, -buffer agents, and analgesic agents may be added. If necessary; coloring' agents, preservativ-es, perfumes, seasoning agents, sweetening agents,'and other medicines may also be *added to the desired preparations- during" the treatment of schizophrenia.
. . :

[0099] The amourit of Tiotropium bromide solvate or pharmaceutically acceptable salt thereof contained in. a pharmaceutical composition for reducing cholesterol according to the present invention is not specifically restricted; however, the dose should be sufficient to treat; ameliorate, or reduce the condition. For example, Tiotropium bromide 'solvate may be present in an amount of about 1%.to about 70%.
[0100] The dosage-of a pharmaceutical composition for reducing cholesterol according to the present invention will depend on the method of use, the age, sex, weight and=condition of the patient.
Typically, about 1 mg to 200 mg of Tiotropium bromide solvate may be containea.. in an administration unit form, preferably a l'O mg tablet. [01011 Having described the inveiztion with reference to certain preferred embodiments, other embodiments will become apparent to one skilled in the art from consideration of the specification. The invention is further defined by r=eference to the following examples describing in detail the process.andcompositions of the invention. It will -be apparent to those skilled in the art that-many modifications, both to materials and methods, may be practiced without departing from the scope of the invention. EXAMPLES

Column: CPS Hypersil; 5 m.; 250.x 4.6 mm.
Mobile phase: Eluent A: 3-mL Perchloric acid 70%w/v in 1000 mL of water Eluent B: Acetonitrile Gradient : Time (min) Mobile Phase AM
Mobile Phase B (~) 35 . 5C
50 40 3'-.30 Flow: = 1.8 ml/min ' Run time: 40 min. Column temperature: 25 C. .
Detector: = W at 240nm.
injection volume: 5 l.
Sample preparation: Tiotropium bromide (1mg/mL in mobile phase) Diluent: Acetonitrile 0.1% V/V; acetic acid in water (50:50 V/V) Post time: _ 5 min.=
-in these.conditions:
Retention time tiotropium bromide: about 3.63 min:
Retention time=dithienylglycolic acid: about 5.4 min Detection limit: 0.005%.

Stability tests of Tiotropium bromide [0102] TABLE 1: Tiotropium Ethanolate dry: stability at 4 C
Time Dithienylglyaolic acid 11 days 20 days - 1 month 1-2 months 0.04%
3 months -[0103] TABLE 2: Tiotropium Ethanolate dry: stability at r.t.
Time Dithienylglycolic ac3.d 11 days -20 days -l month -2 mon.ths 0. 0 4%
-. . 3 month [0104]- TABLE 3: Tiotropium monohydrate micronized: stability at 4 C. Time Dithienylglycolic acid 0.49% . . =
11 days 0.49%
20 days 0. 6 3%
1 month 0_7% 2 months 0=.82%

3 months 0.5% [0105] TABLE 4:,Tiotropium monohydrate micronized: stability-at 25QC. .
Ti.me Dithienylglycolic acid , 0.49% 11 days 0.73%
= .
20 days 0.58~
1 month 0.65%
2 months. 0.72%
3 months 0.58% '.

[0106] TABLE 5: Tiotropium ethanolate micronized: stability at 4oC = = . ' Tim.e Dithienylglycblic acid .
2 -months -3 months - =

Example 1:-Analysis of the SPIRIVA HandiHaler . capsules [0107]. The capsule that were analyzed were part of Lot 4089,66, expiry date 'May 2 0'05 . [0108] 50 mg of sample was dissolved in 50 ml of diluent. The solution was injected, into the chromatographic syst-em equipped with a suitable .injection device - as -blank (as .. Diluent) '. The analysis showed that Tiotropium bromide had a purityof' 98.94$' area by HPLC,, and a content of 0.77% of dithisnylgly.co3.ic acid.
Example 2: Preparation of crude Tiotropium.bromide [.0109] 0.52 g. of N-demethyl tiotropium (1.39 mmol.) * was suspended in 5.23 mL of CH3CN under nitrogen.
[0110] 1.35 g of CH3Br 50% 'w/w solution in CH3CN (-0.0071 mol) were loaded, and the suspension was left under stirring at 22 C
for 12 hours. The product was filtered and washed with 1m1 of.
CH3CN.
[0111] 572 mg of wet Tiotropium bromide were obtained (HPLC
purity 99.89%, dithienylglycolic acid not detected)..
Example 3:. Preparation of Tiotropium bromide [0112]= 4.96g=of N-demethyl tiotropium (13.2 mmol) were loaded in a flask under nitrogen' with 49.6mL of CH3=CN. A suspension was' obtained. 12'.61g of CH3Br 50% w/w -CH3CN sol:uti.on:= (0.066 mol) were loaded.
[01131 The suspension was left under stirring at-22 -C for 64 hours. The product was filtered and washed'with 2mL of CH3CN.
[0114] 6.93g of wet Tiotropium were obtained, and dried under vacuum'at 45 C for 22h (residual pressu=re 4 mbar). 5.9 g of dry product (purity 99.8%, dithienyiglycolic acid-not detected) were obtained.
-Example 4: Crystallization of Tiotropium bromide from absolute ethanol [0115] Tiotropium bromide (1.00' g) was dissolved in absolute ethanol (65 ml) at 78 C. The solution.was heated to 78' C for ..about = 3 0 min, and theri was cooled to 22 C in - at least 6 hours..
The obtained suspension was maintained at 22 C for at least 3 hours, and then was filtered on a sintered glass -funnel, 'and*the solid was washed two times with absolute ethanol -(2 x 1.0 ml)., The= solid 'was dried for 30 min. at 22 C under N2 flow, and= then =for 9 hours at 60 C = under reduced pressure (17 mbar). 0.66 g of Tiotropium bromide(purity .99.68%, dithienylglycolic 'acid-0.01%) were obtained. Example 5: Crys'tallization of Tiotropium bromide=from a-mixture of ethanol and acetic acid [0116] Crude Tiotropium bromide 1.18.6 g) was suspended in ethanol-96%/CH3COOH 98/2 (558 ml). The suspension was heated to 65/70 C until a-solution was obtained, -and then was cooledto 55 C -in at. least 3 hours. and at 0 5 C- in at least 3 hours. The obtained suspension was maintained at 0f5 C for at least ~6 hours, and then was filtered on a sintered glass funnel, and the solid was washed two times with ethanol 96%/CH3COOH 98/2.(3 x 10.0'ml).
The solid was dried for 20.min. at 45 C under reduced.pressure (4 mbar). 16.04 g of Tiotropiumbromide was obtained (purity 99.9%, dithienylglycoTic acid -not detected).
Example 6: Crystallization 'of Tiotropium bromide -from . a. mixture of ethanol and acetic acid -[0117] Crude Tiotropium bromide *(10 g) is suspended in.ethanol 96%/CH3COOH 98/2 (50, ml).The suspensiozi is heated to 65/70. C.
urntil a solution is. obtairi-ed,. and then it cooled, to 55 C in at least 3 hours and at 0 5 C in, at' least 3 hours. 'The obtained-suspension- is maintained',at 0 5 C for at least -6 hours, and then it is filtered on=a sintered glass funnel, and the -solid -is washed two times with ethanol 96$/CH3COOH 98/2 .(3 x 5 ml). The solid is dried for 20 min. at 45 C under reduced, pressure (4 mbar). 8 g of Tiotropium brom'ide isobtained'. (purity' 99.8%, dithienylglycolic acid-0.13%).
Example 7: 'Crystallization of Tiotropium bromide froin 'a mixture of ethanol and-acetic acid 101183 Crude -Tiotropiurct bromide (10 g) is -suspended in ethariol -96%/CH3COOH 98/2 (100 ml.). The suspension- is heated to 65/70 C
until a solution is obtaa.ned,. and then it is 'cooled to 55 C, in at least 3 hours and at 0 5 C in at least 3 hours. The obtained suspension is maintained at 0 5 C for at-:least *6 hours, and then it is filtered=on a sintered glass funnel,.and..the solid' is washed two times with ethanol 96%/CH3COOH 98/2 (3 x 5 ml). Thesolid is .dried for 20 min.. at 45 C under reduced pressure (4 mbar). 8.5 g of Tiotropium bromide is obtained (purity 99.9%, dith3:enylglycolic acid 0.03%). .

Example 8: Crystallization of Tiotropium bromide=from a mixture of-ethanol and acetic acid .
[0119] Crude Tiotropium bromide (10 g) is suspended in ethanol .96%/CH3COOH .98/2 .(200 ml). The suspension is heated to 65/70 C
until a.solution is obtained, and then it is cooled to 55 C in at=
].east 3= hours 'and at 0 5 C 'in at least 3 hour=s. The obtained .suspension is maintained at 0t5 C for at least 6 hours, and then it is filtered on a sintered glass funnel, and. the soli=d i-si washed two times with ethanol 96%/CH3COOH 98/2: (3 x 5 ml).. The solid 'is dried for= 20 min. at' 45 C under reduced pr.essure. (4 mbar): 8.2 g of Tiotropium bromide is obtained (purity 99.9%;
dithienylglycolic acid 0.02%): =
Example 9: Crystallization of Tiotropium bromide ftom a mixture of methanol and acetic acid=. [0120] Crude Tiotropium bromide (10 g) is suspended in methanol 96%/CH3COOH 98/2 (200 ml). The suspension-is heated to 60/65 C until a solution isobtained, and then it is cooled to 45 C in at least 3 hours- and at 0t5 C in, at least 3 hours. The obtained suspen sion. is maintained at 0 5 C for at least 6 hours, ,and then it 'is filtered on, a. sintered glass funnel, and the-solid.
is washed 'two times with methanol 96%/CH3QoOH 98/2 (3 x 5 ml) .
The solid is dried for 20 min. at 45 C under reduced pressure (4 mbar). 8.2 g of Tiotropium bromide is obtained (purity 99.9%, dithienylglycolic acid0.02%).
Example 10: Crystallization of Tiotropium bromide from a mixture of methanol, acetone and acetic acid ' [0121] Crude Tiotropiuin bromide=(10 g) is suspended in mixture' of inethanol/acetone/CH3COOH 73.5/24.5/2= (50 ml): The suspension is heated to 60/65 C until a solution-is obtained, aiad then it,is.
cooled to 45 C in at least 3 hours and at 0t5 C in'at least 3 hours'. The= obtained suspension is maintained at 0 5 C forat least 6 hours, and then it is filtered on a sintered glass funnel, and the solid is washed tw, times with methanol 96%/CH3COOH 98/2 .(3 x 5 ml) . The solid is dried for 2a = m.in. at 45 C = under reduced pressure (4 mbar) .'.8 . 2 g of Tiotr.dpium bromide is obtained (purity 99.9%, dithienylglycolic acid 0..05%).
Example 11: Crystallization of Tiotropium bromide from a mixture of methanol, acetone and acetic-acid.
[0122] Crude Tiotropium bromide (10 g) is suspended in mixture of inethanol/acetone/CH3COOH 24.-5/73.5/2 (50 .m2). The suspension is heated to 60/65 C until a solution is obtained,.and then it is cooled to 45 C in at least 3 hours and at . 0 5 C in at. 3.east 3 hours. The obtained suspension is rnaintained at 0 5 C for. at least 6 hours; and then it is filtered ..on a sintered. glass funnel, and the solid is washed two times -with= methanol 96%/CH3COOH 98/2 '(3 x 5. ml) . The solid is dried for 20 min. -at 45 C under reduced pressure. 8.2- g of '.Tiotropium bromide is obtained' (purity 99.9.%, dithienylglycolic aci:d 0.05%)..
Example 12: Crystallization of Tiotropium bromide from 'a-mixture of methanol,. acetone and acetic.acid .[0123.] Crude Tiotropium bromide (10 g): is suspended in mixture of inethanol/acetone/CH3COOH_24.5/7'3.5/2 (100 ml)_ The-suspension is heated to,60/6-5 C-until a solution is obtained, and then it is cooled to 45 C in at least 3 hours and at 0 5 C in at least 3-hours. The. obtained suspension is maintained at -0+-5 C* for at least .6 hours, and then it is filtered on a sintered glass funnel, and the. solid is washed two times with' methanol 96%/CH3COOH 98/2 (3 x 5 ml) : The solid 'is dried for 20 min. at 45 C under reduced pressure. 7.7 g of Tiotropium bromide a.s-obtained (purity 99.9%, dithienylglycolic acid 0.03%).
Example 13: Crystallization of-Tiotropium bromide from a mixture of methanol, acetone and acetic acid 40124] Crude Tiotropium-bromide (10 g)' is suspended in mixture of inethanol/acetone/CH3COOH 24.5/73:5/2' (300 ml) . .-The 'suspension is heated to 60/65 C until a solution is obtained, arid then it is cooled to. 459C in at least 3 hours and at 0 5 C in at least 3 hours. The obtained suspension is maintained at.0 5 C for at 'least 6 hours, and thery - it is filter.ed. on a sinte.red glass funnel, and the solidi.s washed two times with methanol . 96%%CH3COOH 98/2 (3 x 5 ml) . The solid is dri.ed for 20 min. at 45 C under reduced pressure. 5.1 g of Tiotropium 'bromide is =obtained (purity 99.9%, dithi:enylglycolic acid not detectable)..
Example.14: -Crystallization of Tiotropium bromide from.a mixture.
of n-propanol, water and aceta.c acid ' .[0125]. Crude Tiotropium bromide (10 g) is suspended in n-propanol 93/ water 5/ CH3COOH 2 (200 ml). The suspension is~
heated to-60/65 C until a solution is obtained, and then it is cooled' to 45 C- in at least 3'hours and at 0 5 C in at least 3 hours: The obtained suspension.is maintained at 0 5 C for at least 6 hours, and then it' is filtered on a=sintered gla'ss funnel, and the solid is washed 'two times with n-propanol 96%/CH3COOH' 98/2 (3 x 5 ml).. The scilid is dried for 20_ min. at 45 C under reduced pressure. 8.2 g of Tiotropium bromide is obtained (purity 99.9%, 'dithienylglycolic acid 0.02%). Example 15:
Crystallization. of Tiotropium bromide f=rom a mixture of n-propanol and acetic .acid- [0126] Wet crude.Tiotropium bromide (10g)'is suspended in n-propanol - 98/ . CH3COOH 2 (8500 ml ). The suspension is heated to 60/65 C until a solution is obtained, and then it is..cooled to, 45 C iri at least 3 hours and at 0f5 C in at least3 hours.. The obtained suspension is maintained at.0 5 C for.at least 6hours, and then it is filtered on a sintered glass funnel, and the solid=
is washed two times with n-propanol 9 6%/ CH3COOH 9 8/ 2 (3 x 5 ml).
The solid is dried for 20 min. at 45 C under reduced pressure.
4.2 g 'of Tiotropium bromide is obtained (purity 99.9%, dithienylglycolic=acid not-detectable).
Example 16: Crystall'i2ation of Tiotropium bromide from a mixture.
of n-propanol and acetic acid [0127] Wet crude Tiotropium bromide (10 g) is suspended in n-propanol 98/ CH3COOH 2 (50 ml). The suspension is heated to .60/65 C until a solution is obtained, and then it is cooled to 45 C in at least .3 hours and at 0 5 C ' in at least 3 hours. The obtained suspension*is maintained at '0 5 C for at least 6 hours, and theri it is filtered on a sintered glass funnel,.and the solid is washed two times with n-propanol 96%/CH3COOH 9 8/ 2 0. x 5 ml ).
The solidis dried-for 20 min.- at 45 C under reduced pressure.
8.2 g of Tiotropium bromide is obtained (purity 99.9%, dithienylglycolic acid 0.08). . . . Example 17: Crystallization of Tiotropium-bromide -from a=mixture of n-propanol and acetic acid .
[0128] Wet crude Tiotropium bromide (10 g) is suspended in n-propanol 98/ CH3.COOH 2 (100 ml). The suspension =is heated 'to 6.0/65 C until a solution is obtained, and then* it is cooled ta 45 C in at least =3 '.hours and at 0t5 C= in at least 3 hours. The obtained suspension is maintained at 0 5 C for at ieast-6. hours,-and then it is filtered on'a sintered glass funnel., and the sol.id,.
is washed two times with ri-propanol 96%/CH3COOH 98/2 (3 'x 5 ml).
The solid is dried for 20 mi.n.. at 45 C under reduced pr.es-sure.
4.2 g of Tiotropium bromide, is obtained (purity. 999.9%, dithienylglycolic acid 0.03). .
.Example 18: Preparation of Tiotropium bromide from: Ac'OH/MeOH/n-Heptan 7/2/2.3 [0129] Tiotropium bromide (1.00 g) was dissolved at .45 C with a mixture 7/2 (V/V) of -acetic acid/methanol (11 ml), the solution was heated to 45 C.for 1.5 hours and n-heptane'(2.75 ml) was then added drop-wise in at least.' 20 min.. The obtained solution was heated to 45 C for one hour Ano solid formation=
observed) ; was cooled to 23..5 C ' in at least 3 hours and the suspension was maintained at .23.5 C for at least 3'hours.. After=
filtration on a sintered glass funnel, the solid'was washed -three times with 3.0 mL of-n-heptane. Drying for 16 hours at 60 C under reduced pressure (18 . mbar) , yielded'0. 67 g of.Tiotropium biomide (purity 99.9%., dithienylglycolic acid 0.03). ' Example 19: Micronization process [0130]. Tiotropium Bromide was micronized to obtain P.S.D
. =
target of:

Min. 80%<5 . 84-~un Min. 70% between 0.6 and 10 microns The micronizer in use was a Jet-mill -MC 5-0. (made by Micro-Macinazionne).. 32 05' angle nozzles.we=re installed.

Nitrogen was used as-the micronization gas. Micronizatiori air Pressure'was 10 bars.

Feed rate was 0.2 kg/hr.
The micronized Tiotropium bromide obtained by the:-above, process has.a PSD value:
80%5'5. 84pm- .
93.76% between 0..6 and 10 microns.

Claims (57)

1. Tiotropium bromide solvate of the following formula:

2. The Tiotropium bromide solvate of claim 1, wherein the Tiotropium bromide has a purity ranging from about 99% to about 100%.

3. The Tiotropium bromide solvate of claim 2, wherein the Tiotropium bromide has a purity ranging from about 99.5% to about 100%.

4. The Tiotropium bromide of solvate claim 3, wherein the Tiotropium bromide has a purity ranging from about 99.7% to about 100%.

5. The Tiotropium bromide solvate as defined in any of claims 1 to 4, wherein the Tiotropium bromide includes less than about 0.15% of 2,2-dithienyl glycolic acid.

6. A Tiotropium bromide solvate with less than about 0.15%
of 2,2-dithienyl glycolic acid.

7. The Tiotropium bromide solvate of claim 6, wherein the Tiotropium bromide contains from about 0.15% to the detection limit of an HPLC method of 2,2-dithienyl glycolic acid.

8. The Tiotropium bromide solvate of claim 7, wherein the Tiotropium bromide contains less than about 0.05% of 2,2-dithienyl glycolic acid.

9. The Tiotropium bromide solvate of claim 8, wherein the Tiotropium bromide contains from about 0.05% to the detection limit of an HPLC method of 2,2-dithienyl glycolic acid.

10. The Tiotropium bromide solvate of claim 6, wherein the Tiotropium bromide contains less than about 0.02% of 2,2-dithienyl glycolic acid.

11. The Tiotropium bromide solvate of-claim 10, wherein the Tiotropium bromide contains from about 0.02% to the detection limit of an HPLC method of 2,2-dithienyl glycolic acid.

12. The Tiotropium bromide solvate of claim 6, wherein the Tiotropium bromide contains less than about 0.01% of 2,2-dithienyl glycolic acid.

13. The Tiotropium bromide solvate of claim 1-2, wherein the Tiotropium bromide contains from about 0.01% to the detection limit of an HPLC method of 2,2-dithienyl glycolic acid.

14. A stable Tiotropium bromide solvate.

15. The stable Tiotropium bromide solvate of claim 14, wherein the level of 2,2-dithienyl. glycolic acid does not increase to more than 0.15%, when maintained at a temperature ranging from about 4°C to about 30°C, for at least about two months.

16. The Tiotropium bromide solvate as defined in any of claims 1 to 13, wherein the Tiotropium bromide solvate is stable.

17. The Tiotropium bromide solvate as defined in any of claims 1 to 15, wherein the Tiotropium bromide solvate is selected from the group consisting of an acetic acid solvate and an alcohol solvate.

18. The solvate of claim 17, wherein the alcohol solvate is a C1-8 alcoholate.

19. The solvate of claim 18, wherein the C1-8 alcoholate is C1-6 alcoholate.

20. The solvate of claim 19, wherein the C1-6 alcoholate is C2-5 alcoholate.

21. The solvate of claim 20, wherein the C1-5 alcoholate is C1-4 alcoholate.

22. The solvate of claim 21, wherein the C1-4 alcoholate is selected from the group consisting of methanolate, ethanolate, sopropanolate, n-propanolate and n-butanolate.

23. The solvate of claim 22, wherein the C1-4 alcoholate is selected from the group consisting of methanolate, ethanolate or n-propanolate.

24. An HPLC method comprising:
(a) combining a Tiotropium bromide sample with a mixture of acetonitrile:acetic acid in water in a ratio of about 0.1%:99.9%, to obtain a solution;
(b) injecting the solution into a column;
(c) eluting the sample from the column at about 3.63 min using an eluent mixture comprised of a first eluent and a second eluent, the first eluent comprising perchloric acid:water in a ratio of 7:3 and the second eluent comprising acetonitrile; and (d) measuring the 2,2-dithienyl glycolic acid content in the relevant sample with a UV detector.

25. The HPLC method of claim 24, wherein the column is a 250×4.6 mm ×0.5 µm CPS Hypersil (or similar) column.

26. The method of claim 24, wherein the ratio of the first and second eluents varies over time.

27. The method of claim 26, wherein at the time 0 minutes, the eluent contains 70% of the first eluent and 300% of the second eluent.

28. The method of claim 26, wherein at the time 23 minutes, the eluent contains 55% of the first eluent and 45% of the second eluent.

29. The method of claim 26, wherein at the time 30 minutes, the eluent contains 50% of the first eluent and 50% of the second eluent.

30. The method of claim 26, wherein at the time 35 minutes, the eluent contains 50% of the first eluent and 50% of the second eluent.

31. The method of claim 26, wherein at the time 40 minutes, the eluent contains 35% of the first eluent and 65% of the second eluent.

32. The method of claim 26, wherein at the time 41 minutes, the eluent contains 70% of the first eluent and 30% of the second eluent.

33. The method of claim 24, wherein the 2,2-dithienyl glycolic acid content is measured at a wave length of 240 nm.

34. A process for preparing stable Tiotropium bromide solvate having a purity of at least 99% and containing less than about 0.15% of 2,2-dithienyl glycolic acid, comprising crystallizing Tiotropium bromide from a solvent system comprising an organic acid, wherein the ratio of Tiotropium bromide to the solvent system is at least about 1 to about 5.

35. A process for preparing stable Tiotropium bromide solvate having a purity of at least 99% and with less than about 0.05% of 2,2-dithienyl glycolic acid as measured by HPLC, comprising crystallizing Tiotropium bromide from a solvent system comprising an organic acid, wherein the ratio of Tiotropium bromide to the solvent system is at least about 1 to about 10.

36. A process for preparing stable Tiotropium bromide solvate having a purity of at least 99% and with less than about 0.02% of 2,2-dithienyl glycolic acid, comprising crystallizing Tiotropium bromide from a solvent system comprising an organic acid, wherein the ratio of Tiotropium bromide to the solvent system is of at least about 1 to about 20.

37. A process for preparing stable Tiotropium bromide solvate having a purity of at least 99% and with less than about 0.01% of 2,2-dithienyl glycolic acid, comprising crystallizing Tiotropium bromide from a solvent system comprising an organic acid, wherein the ratio of Tiotropium bromide to the solvent system is of at least about 1 to about 30.

38. The process as defined in any of claims 34 to 37, wherein the organic acid is selected from the group consisting of

39 trifluoroacetic acid, tartaric acid, maleic acid, propionic acid, oxalic acid, p-toluen sulphonic acid, methan sulphonic acid and acetic acid.

39. The process of claim 38, wherein the organic acid is acetic acid.

40. The process as defined in any of claims 34 to 37, further comprising cooling the solution to obtain a suspension.

41. The process of claim 40, wherein the solvent system further comprises an alcohol.

42. The process of claim 41, wherein the alcohol is a C1-8 alcohol.

43. The process of claim 42, wherein the C1-8 alcohol is C1-6 alcohol.

44. The process of claim 43, wherein the C1-6 alcohol is C1-5 alcohol.

45. The process of claim 44, wherein the C1-5 alcohol is C1-4-alcohol.

46. The process of claim 45, wherein the C1-4 alcohol is selected from the group consisting of methanol, ethanol, isopropanol, n-propanol and n-butanol.

47. The process of claim 46, wherein the C1-4 alcoholate is selected from the group consisting of methanol, ethanol and n-propanol.

48. The process as defined in any of claims 34 to 47, further comprising heating the combination of Tiotropium bromide and the solvent system.

49. The process of claim 48, wherein the heating is to a temperature ranging from about 60°C to about 78°C.

50. The process of claim 40, wherein the solution is cooled to a temperature ranging from about 25°C to about O°C.

51. The process of claim 50, wherein the cooling is done over a period of about 4 to about 10 hours.

52. The process of claim 40, wherein the process further comprises recovering the crystallized product.

53. A pharmaceutical composition comprising a tiotropium bromide solvate as defined in any of claims 1 to 23, and at least one pharmaceutically acceptable excipient.

54. Use of a tiotropium bromide solvate as defined in any of claims 1 to 23 for the manufacture of a composition for the treatment of asthma or chronic obstructive pulmonary disease.

55. The triotropium bromide solvate as defined in any of claims 1 to 23, wherein the tiotropium bromide solvate is micronized.

56. A process for preparing Tiotropium bromide including less than about 0.15% area by HPLC of 2,2-dithienyl glycolic acid comprising the steps of:
(a) obtaining one or more samples of one or more Tiotrpium bromide batches;
(b) measuring a level of 2,2-dithienyl glycolic acid in each of the samples by HPLC;
(c) selecting the Tiotropium bromide batch that comprises a level of 2,2-dithienyl glycolic acid of less than about 0.15% area by HPLC; and (d) using the selected batch to prepare a Tiotropium bromide comprising less than about 0.15% area by HPLC of 2,2-dithienyl glycolic acid.

57. The process of claim 56, further comprising purifying the samples prior to measurement.