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`CEPH—4604/CP391 D US
`PATENT APPLICATION
`TRANSM ITTAL
`(0nlyfornew nonprovisional applications under 37CFR 1.53(b))
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`FRESENIUS KABI 1003-OOO1
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`FRESENIUS KABI 1003-OOO2
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`CEPH-4604 CP391D
`
`PATENT
`
`BENDAMUSTINE PHARMACEUTICAL COMPOSITIONS
`
`CROSS REFERENCE TO RELATED APPLICATIONS
`
`This application is a continuation of U.S. Application No. 13/719,409, filed
`
`December 19, 2012, which is a continuation of U.S. Application No. 13/654,898, filed
`
`October 18, 2012, now U.S. 8,461,350, which is a continuation of U.S. Application No.
`
`11/330,868, filed January 12, 2006, now U.S. 8,436,190, which claims the benefit ofU.S.
`
`Provisional Application No. 60/644,354, filed January 14, 2005, the entireties of which
`
`are incorporated herein for all purposes.
`
`FIELD OF THE INVENTION
`
`The present invention pertains to the field of pharmaceutical compositions for the
`
`treatment of various disease states, especially neoplastic diseases and autoimmune
`
`diseases. Particularly, it relates to pharmaceutical formulations comprising nitrogen
`
`mustards, particularly the nitrogen mustard bendamustine, e.g., bendamustine HCl.
`
`BACKGROUND OF THE INVENTION
`
`The present invention claims the benefit of and priority to US Serial No.
`
`60/644,354, filed January 14, 2005, entitled, “Bendamustine Pharmaceutical
`
`Compositions,” which is incorporated herein by reference in its entirety, including figures
`
`and claims.
`
`The following description includes information that may be useful in
`
`understanding the present invention. It is not an admission that any such information is
`
`prior art, or relevant, to the presently claimed inventions, or that any publication
`
`specifically or implicitly referenced is prior art.
`
`Because of their high reactivity in aqueous solutions, nitrogen mustards are
`
`difficult to formulate as pharmaceuticals and are often supplied for administration in a
`
`lyophilized form that requires reconstitution, usually in water, by skilled hospital personal
`
`prior to administration. Once in aqueous solution, nitrogen mustards are subject to
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`CEPH-4604 CP39 l D
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`PATENT
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`degradation by hydrolysis, thus, the reconstituted product should be administered to a
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`patient as soon as possible after its reconstitution.
`
`Bendamustine, (4- {5 - [Bis(2-chloroethyl)amino] - l -methyl-2-benzimidazolyl}
`
`butyric acid, is an atypical structure with a benzimidazole ring, whose structure includes
`
`an active nitrogen mustard (see Formula I, which shows bendamustine hydrochloride).
`
`O
`N
`cl/fiN
`Clf \E:Er\{\>—\_>-OH .HC|
`
`Formula I
`
`l0
`
`Bendamustine was initially synthesized in 1963 in the German Democratic
`
`Republic (GDR) and was available from l97l to l992 in that location under the name
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`Cytostasan®. Since that time, it has been marketed in Germany under the tradename
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`Ribomustin®. It has been widely used in Germany to treat chronic lymphocytic
`
`leukemia, Hodgkin’s disease, non-Hodgkin’s lymphoma, multiple myeloma, and breast
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`l5
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`cancer.
`
`Due to its degradation in aqueous solutions (like other nitrogen mustards),
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`bendamustine is supplied as a lyophilized product. The current lyophilized formulation of
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`bendamustine (Ribomustin®) contains bendamustine hydrochloride and mannitol in a
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`sterile lyophilized form as a white powder for intravenous use following reconstitution.
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`The finished lyophilisate is unstable when exposed to light. Therefore, the product is
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`stored in brown or amber-colored glass bottles. The current lyophilized formulation of
`
`bendamustine contains degradation products that may occur during manufacturing of the
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`drug substance and/or during the lyophilization process to make the finished drug product.
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`Currently bendamustine is formulated as a lyophilized powder for injection with
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`100 mg of drug per 50 mL vial or 25 mg of drug per 20 mL vial. The vials are opened and
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`reconstituted as close to the time of patient administration as possible. The product is
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`reconstituted with 40 mL (for the 100 mg presentation) or l0 mL (for the 25 mg
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`presentation) of Sterile Water for Injection. The reconstituted product is further diluted
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`20
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`25
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`FRESENIUS KABI 1003-OOO4
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`CEPH-4604 CP391D
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`PATENT
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`into 500 mL, q.s., 0.9% Sodium Chloride for Injection. The route of administration is by
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`intravenous infusion over 30 to 60 minutes.
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`Following reconstitution with 40 mL Sterile Water for Injection, Vials of
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`bendamustine are stable for a period of 7 hours under room temperature storage or for 6
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`days upon storage at 2-8°C. The 500 mL admixture solution must be administered to the
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`patient within 7 hours of Vial reconstitution (assuming room temperature storage of the
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`admixture).
`
`The reconstitution of the present bendamustine lyophilized powder is difficult.
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`Reports from the clinic indicate that reconstitution can require at least fifteen minutes and
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`may require as long as thirty minutes. Besides being burdensome and time-consuming for
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`the healthcare professional responsible for reconstituting the product, the lengthy exposure
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`of bendamustine to water during the reconstitution process increases the potential for loss
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`of potency and impurity formation due to the hydrolysis of the product by water.
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`Thus, a need exists for lyophilized formulations of bendamustine that are easier to
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`reconstitute and which have a better impurity profile than the current lyophilate
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`(lyophilized powder) formulations of bendamustine.
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`German (GDR) Patent No. 34727 discloses a method of preparing oa-[5-bis-(B-
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`chloroethyl)-amino-benzimidazolyl-(2)]-alkane carboxylic acids substituted in the 1-
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`position.
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`German (GDR) Patent No. 80967 discloses an injectable preparation of y-[1-
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`methyl-5-bis-(B-chloroethyl)-amino-benzimaidazolyl-(2)]-butric acid hydrochloride.
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`German (GDR) Patent No. 159877 discloses a method for preparing 4-[l-methyl-
`
`5-bis (2-chloroethyl) amino-benzimidazolyl-2)-butyric acid.
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`German (GDR) Patent No. 159289 discloses an injectable solution of
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`10
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`15
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`20
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`25
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`bendamustine.
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`Ribomustin® bendamustine Product monograph (updated 1/2002)
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`http://www.ribosepharm.de/pdf/ribomustin bendamustin/productmonographpdf provides
`
`information about Ribomustin® including product description.
`
`Ni et al. report that the nitrosourea SarCNU was more stable in pure tertiary
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`30
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`butanol than in pure acetic acid, dimethyl sulfoxide, methylhydroxy, water or in
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`TBA/water mixtures (Ni et al. (2001) Int]. J. Phamaceutics 226:39-46).
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`-3-
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`FRESENIUS KABI 1003-0005
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`CEPH-4604 CP391D
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`PATENT
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`Lyophilized cyclophoshamide is known in the art see e. g., US Patent Nos.
`
`5,418,223; 5,413,995; 5,268,368; 5,227,374; 5,130,305; 4,659,699; 4,537,883; and
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`5,066,647.
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`The lyophilized nitrogen mustard Ifosfamide is disclosed in International
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`Publication No. WO 2003/066027; US Pat. Nos. 6,613,927; 5,750,131; 5,972,912;
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`5,227,373; and 5,204,335.
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`Teagarden et al. disclose lyophilized formulations of prostaglandin E-1 made by
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`dissolving PGE-1 in a solution of lactose and tertiary butyl alcohol (US Pat. No.
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`5,770,230).
`
`SUMMARY OF THE INVENTION
`
`The present invention is directed to stable pharmaceutical compositions of
`
`nitrogen mustards, in particular lyophilized bendamustine and its use in treatment of
`
`various disease states, especially neoplastic diseases and autoimmune diseases.
`
`An embodiment of the invention is a pharmaceutical composition of bendamustine
`
`containing not more than about 0.5% to about 0.9% (area percent of bendamustine) HP1,
`
`as shown in Formula II,
`
`Ho/fl
`r”r:”w}oH
`"{
`
`Cl
`
`Formula II
`
`at the time of release or where the HP1 is the amount of HP1 present at time zero after
`
`reconstitution of a lyophilized pharmaceutical composition of bendamustine as described
`
`herein. In a preferred embodiment is a pharmaceutical composition of bendamustine
`
`containing not more than about 0.5% (area percent of bendamustine) HP1, preferably not
`
`more than about 0.45%, more preferably not more than about 0.40%, more preferably not
`
`more than about 0.35%, even more preferably not more than 0.30%.
`
`Another embodiment of the invention is a lyophilized preparation of bendamustine
`
`containing not more than about 0.1 % to about 0.3 % bendamustine dimer as shown in
`
`Formula III at release or at time zero after reconstitution
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`10
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`15
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`20
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`25
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`FRESENIUS KABI 1003-0006
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`CEPH-4604 CP39 l D
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`PATENT
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`Ho/fl
`
`HO/\/N N\
`
`N
`
`N
`
`o
`
`0 OJ N\%\—>—OH
`
`Formula III.
`
`Yet another embodiment of the invention is a lyophilized preparation of
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`5
`
`bendamustine containing not more than about 0.5%, preferably 0. 15% to about 0.5%,
`
`bendamustine ethylester, as shown in Formula IV at release or at time zero after
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`reconstitution
`
`CI
`
`COOCHZCH3
`R
`(NQIWN
`
`I
`
`Formula IV.
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`10
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`Yet another embodiment of the invention is a lyophilized preparation of
`
`bendamustine wherein the concentration of bendamustine ethylester (Formula IV) is no
`
`more than 0.2%, preferably 0. l%, greater than the concentration of bendamustine
`
`ethylester as found in the drug substance used to make the lyophilized preparation.
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`15
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`In another embodiment of the invention is a lyophilized preparation of
`
`bendamustine containing not more than about 0.5% to about 0.9% (area percent of
`
`bendamustine) HPl at the time of drug product release. In a preferred embodiment is a
`
`lyophilized preparation of bendamustine containing not more than about 0.50% (area
`
`percent of bendamustine) HPl , preferably not more than about 0.45%, more preferably
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`20
`
`not more than about 0.40%, more preferably not more than about 0.35%, even more
`
`preferably not more than 0.30%. An aspect of this embodiment is lyophilized
`
`preparations of bendamustine containing not more than about 0.5% to about 0.9%,
`
`preferably 0.5%, (area percent of bendamustine) HPl at the time of release of drug
`
`product where the lyophilized preparation is packaged in a vial or other pharrnaceutically
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`25
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`acceptable container.
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`FRESENIUS KABI 1003-0007
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`CEPH-4604 CP39 l D
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`PATENT
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`In yet another aspect of the invention, the lyophilized preparations of
`
`bendamustine are stable with respect to the amount of HP1 for at least about 6 months,
`
`preferably 12 months, preferably 24 months, to about 36 months or greater when stored at
`
`about 2° to about 30°. Preferred temperatures for storage are about 5° C and about room
`
`temperature.
`
`Another embodiment of the invention is a pharmaceutical dosage form that
`
`includes a pharmaceutical composition of bendamustine containing not more than about
`
`0.5% to about 0.9% HP1 , preferably not more than about 0.50%, preferably not more than
`
`about 0.45%, more preferably not more than about 0.40%, more preferably not more than
`
`l0
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`about 0.35%, even more preferably not more than 0.30%, where the HP1 is the amount of
`
`HPl present at release or at time zero after reconstitution of a lyophilized preparation of
`
`bendamustine of the present invention. In preferred aspects of the invention, the dosage
`
`form can be about 5 to about 500 mg of bendamustine, about 10 to about 300 mg of
`
`bendamustine, about 25 mg of bendamustine, about 100 mg of bendamustine, and about
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`15
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`200 mg of bendamustine.
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`Yet another embodiment of the invention is a pharmaceutical dosage form that
`
`includes a lyophilized preparation of bendamustine containing not more than about 0.5%
`
`to about 0.9%, preferably 0.5%, HP1. Preferred dosage forms can be about 5 to about 500
`
`mg of bendamustine, about 10 to about 300 mg of bendamustine, about 25 mg of
`
`bendamustine, about 100 mg of bendamustine, and about 200 mg of bendamustine.
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`In still another embodiment, the invention includes a pharmaceutical composition
`
`of bendamustine including bendamustine containing not more than about 0.5% to about
`
`0.9% (area percent of bendamustine), preferably not more than about 0.50%, preferably
`
`not more than about 0.45%, more preferably not more than about 0.40%, more preferably
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`not more than about 0.35%, even more preferably not more than 0.30%, and a trace
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`amount of one or more organic solvents, wherein said HP1 is the amount of HP1 present
`
`at release or time zero after reconstitution of a lyophilized pharmaceutical composition of
`
`bendamustine as disclosed herein. In different aspects of this embodiment, the organic
`
`solvent is selected from one or more of tertiary butanol, n-propanol, n-butanol,
`
`isopropanol, ethanol, methanol, acetone, ethyl acetate, dimethyl carbonate, acetonitrile,
`
`dichloromethane, methyl ethyl ketone, methyl isobutyl ketone, l-pentanol, methyl acetate,
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`FRESENIUS KABI 1003-0008
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`CEPH-4604 CP39 l D
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`PATENT
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`carbon tetrachloride, dimethyl sulfoxide, hexafiuoroacetone, chlorobutanol, dimethyl
`
`sulfone, acetic acid, and cyclohexane. Preferred organic solvents include one or more of
`
`ethanol, methanol, propanol, butanol, isopropanol, and tertiary butanol. A more preferred
`
`organic solvent is tertiary butanol, also known as TBA, t-butanol, tert-butyl alcohol or
`
`tertiary butyl alcohol.
`
`The present invention involves a method for obtaining agency approval for a
`
`bendamustine product, the improvement which includes setting a release specification for
`
`bendamustine degradants at less than about 4.0%, preferably about 2.0 % to about 4.0 %,
`
`(area percent bendamustine) or otherwise to achieve the pharmaceutical compositions
`
`described herein. An aspect of this embodiment is a method for obtaining agency
`
`approval for a bendamustine product which includes setting a release specification for
`
`HPl to be less than or equal to 1.5% (area percent Bendamustine). The bendamustine
`
`product herein contains not more than about 0.5% (area percent of bendamustine) HPl at
`
`release.
`
`Another embodiment is a method for obtaining agency approval for a
`
`bendamustine product, the improvement which includes setting a shelf-life specification
`
`for bendamustine degradants at less than about 7.0%, preferably about 5.0% to about
`
`7.0%, (area percent bendamustine) where the product is stored at about 2°C to about
`
`30°C. Preferred temperatures for storage are about 5°C and about room temperature. The
`
`bendamustine product herein contains not more than about 0.5% (area percent of
`
`bendamustine) HP1 at release.
`
`Another embodiment of the invention is a process for manufacturing a lyophilized
`
`preparation of bendamustine which includes controlling for the concentration of
`
`bendamustine degradants in the final product, such that the concentration of bendamustine
`
`degradants is less than about 4.0%, preferably no more than about 2.0 % to about 4.0 %,
`
`(area percent of bendamustine) at release or otherwise to achieve the pharmaceutical
`
`compositions described herein. The bendamustine product herein contains not more than
`
`about 0.5% to about 0.9%, preferably about 0.5%, (area percent of bendamustine) HP1 at
`
`release.
`
`The present invention discloses a process for manufacturing a lyophilized
`
`preparation of bendamustine which comprises controlling for the concentration of
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`FRESENIUS KABI 1003-0009
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`CEPH-4604 CP39 l D
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`PATENT
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`bendamustine degradants in the final product, such that, at release, the concentration of
`
`HP1 is less than 0.9%, preferably 0.5%, (area percent of bendamustine) and, at the time of
`
`product expiration, the concentration of bendamustine degradants is less than about 7.0%,
`
`preferably no more than about 5.0% to about 70%; wherein said product is stored at about
`
`2°C to about 30°C.
`
`Another embodiment of the invention is a bendamustine pre-lyophilization
`
`solution or dispersion comprising one or more organic solvents where the solution or
`
`dispersions include at least one stabilizing concentration of an organic solvent which
`
`reduces the level of degradation of bendamustine so that the amount of HP1 produced
`
`during lyophilization from about 0 to 24 hours does not exceed about 0.5% to about 0.9%
`
`(area percent of bendamustine) preferably 0.50%, preferably 0.45%, more preferably
`
`0.40%, more preferably 0.35%, even more preferably 0.30%. An aspect of this
`
`embodiment is the lyophilized powder produced from the pre-lyophilization solution or
`
`dispersion.
`
`Still another embodiment of the invention is a bendamustine pre-lyophilization
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`solution or dispersion comprising one or more organic solvents where the solution or
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`dispersions include at least one stabilizing concentration of an organic solvent which
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`reduces the level of degradation of bendamustine so that the amount of bendamustine
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`ethylester produced during lyophilization from about 0 to 24 hours does not exceed about
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`0.5% (area percent bendamustine). An aspect of this embodiment is the lyophilized
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`powder produced from the pre-lyophilization solution or dispersion.
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`Still another embodiment of the invention is a bendamustine pre-lyophilization
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`solution or dispersion comprising one or more organic solvents where the solution or
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`dispersions include at least one stabilizing concentration of an organic solvent which
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`reduces the level of degradation of bendamustine so that the amount of bendamustine
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`ethylester (as shown in Formula IV) produced during lyophilization from about 0 to 24
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`hours is no more than 0.2%, preferably 0.1%, greater than the concentration of
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`bendamustine ethylester as found in the drug substance used to make the pre-
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`lyophilization solution. A preferred organic solvent is tertiary butanol.
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`The invention also discloses methods for preparing a bendamustine lyophilized
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`preparation that includes dissolving bendamustine in a stabilizing concentration of an
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`FRESENIUS KABI 1003-0010
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`

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`CEPH-4604 CP39 l D
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`PATENT
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`alcohol solvent of between about 5% to about 100% (V/V alcohol to form a pre-
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`lyophilization solution; and lyophilizing the pre-lyophilization solution; wherein the
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`bendamustine lyophilized preparation made from such methods contains not more than
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`about 0.5% to about 0.9%, preferably 0.5%, (area percent of bendamustine) HPl as shown
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`in Formula 11, wherein said HPl is the amount of HPl present at release or at time zero
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`after reconstitution of the lyophilized pharmaceutical composition of bendamustine.
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`Other alcohol concentrations include about 5% to about 99.9%, about 5% to about 70%,
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`about 5% to about 60%, about 5% to about 50%, about 5% to about 40%, about 20% to
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`about 35%. Preferred concentrations of alcohol are from about 20% to about 30%.
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`l0
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`Preferred alcohols include one or more of methanol, ethanol, propanol, iso-propanol,
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`butanol, and tertiary-butanol. A more preferred alcohol is tertiary-butanol. A preferred
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`concentration of tertiary-butanol is about 20% to about 30%, preferably about 30%. An
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`aspect of this embodiment is the addition of an excipient before lyophilization. A
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`preferred excipient is mannitol. Preferred pre-lyophilized concentrations of bendamustine
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`l5
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`are from about 2 mg/mL to about 50 mg/mL.
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`In a preferred method for preparing a bendamustine lyophilized preparation,
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`lyophilizing the pre-lyophilization solution comprises i) freezing the pre-lyophilization
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`solution to a temperature below about -40°C, preferably -50°C,
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`to form a frozen solution;
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`ii) holding the frozen solution at or below -40°C, preferably -50°C, for at least 2 hours;
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`iii) ramping the frozen solution to a primary drying temperature between about -40°C and
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`about -10°C to form a dried solution; iv) holding for about 10 to about 70 hours; V)
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`ramping the dried solution to a secondary drying temperature between about 25°C and
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`about 40°C; and Vii) holding for about 5 to about 40 hours to form a bendamustine
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`lyophilized preparation. In a more preferred method lyophilizing the pre-lyophilization
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`solution comprises i) freezing the pre-lyophilization solution to about -50°C to form a
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`frozen solution;
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`ii) holding the frozen solution at about -5 0°C for at least 2 hours to about
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`4 hours; iii) ramping to a primary drying temperature between about -20°C and about -
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`12°C to form a dried solution; iv) holding at a primary drying temperature for about l0 to
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`about 48 hours; V) ramping the dried solution to a secondary drying temperature between
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`about 25°C and about 40°C; and Vi) holding at a secondary drying temperature for at least
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`5 hours up to about 20 hours. A preferred alcohol is tertiary-butanol. A preferred
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`FRESENIUS KABI 1003-0011
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`

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`CEPH-4604 CP39 l D
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`PATENT
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`concentration of tertiary-butanol is about 20% to about 30%, preferably about 30%. An
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`aspect of this embodiment is the addition of an excipient before lyophilization. A
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`preferred excipient is mannitol. Preferred pre-lyophilized concentrations of bendamustine
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`are from about 2 mg/mL to about 50 mg/mL.
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`Another embodiment of the invention is the lyophilized powder or preparation
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`obtained from the methods of preparing a bendamustine lyophilized preparation disclosed
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`herein.
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`The invention also involves bendamustine formulations for lyophilization that
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`include an excipient and a stabilizing concentration of an organic solvent. A preferred
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`formulation includes bendamustine at a concentration of about l5 mg/mL, mannitol at a
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`concentration of about 25.5 mg/mL, tertiary-butyl alcohol at a concentration of about 30%
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`(v/v) and water. Included in this embodiment of the invention are the lyophilized
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`preparations made from such bendamustine formulations.
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`Included in the inventions are methods of treating a medical condition in a patient
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`that involve administering a therapeutically effective amount of a pharmaceutical
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`composition of the invention where the condition is amenable to treatment with said
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`pharmaceutical composition. Some conditions amenable to treatment with the
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`compositions of the invention include chronic lymphocytic leukemia (CLL), Hodgkin’s
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`disease, non-Hodgkin’s lymphoma (NHL), multiple myeloma (MM), breast cancer, small
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`cell lung cancer, hyperproliferative disorders, and an autoimmune disease. Preferred
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`conditions include NHL, CLL, breast cancer, and MM. Preferred autoimmune diseases
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`include rheumatoid arthritis, multiple sclerosis or lupus.
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`Included in the inventions are the use of the pharmaceutical compositions or
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`pharmaceutical preparations of the invention in the manufacture of a medicament for the
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`treatment of a medical condition, as defined herein, in a patient that involve administering
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`a therapeutically effective amount of a pharmaceutical composition of the invention where
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`the condition is amenable to treatment with said pharmaceutical composition.
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`Also included in the invention are methods of treating in which the pharmaceutical
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`compositions of the invention are in combination with one or more anti-neoplastic agents
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`where the antineoplastic agent is given prior, concurrently, or subsequent to the
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`FRESENIUS KABI 1003-0012
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`CEPH-4604 CP391D
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`PATENT
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`administration of the pharmaceutical composition of the invention. Preferred
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`antineoplastic agents are antibodies specific for CD20.
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`Another embodiment of the invention is a lyophilization cycle for producing lyophilized
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`bendamustine preparations of the invention. A preferred lyophilization cycle includes a)
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`freezing to about -50°C over about 8 hours; b) holding at -50°C for about 4 hours; c)
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`ramping to -25°C over about 3 hours; d) holding at -10°C for 30 hours; e) ramping to
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`between about 25°C and about 40°C or higher for about 3 hours; f) holding between about
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`25°C and about 40°C for about 25 hours; g) ramping to about 20°C in 1 hour; h) unloading
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`at about 20°C, at a pressure of 13.5 psi in a pharrnaceutically acceptable container that is
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`hermetically sealed; wherein the pressure is about 150 microns throughout primary drying
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`and 50 microns throughout secondary drying. An aspect of this cycle involves step (e)
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`which is ramped to about 30-35°C for 3 hours and then ramped to 40°C for 5 hours.
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`Another aspect of this embodiment is the lyophilized powered prepared from such
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`lyophilization cycles. A more preferred lyophilization cycle includes i) starting with a
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`shelf temperature of about 5°C for loading; ii) freezing to about -50°C over about 8 hours;
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`iii) holding at -50°C for about 4 hours; iv) ramping to about -20°C over about 3 hours; v)
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`holding at about -20°C for 6 hours; ramping to about -15°C over about 1 hour; vi) holding
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`at -15°C for about 20 hours; vii) ramping to about -15°C over about 1 hour; viii) holding
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`at about -15°C for about 20 hours; ix) ramping to about -12°C over about 0.5 hours; X)
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`holding at about -12°C for about 15.5 hours; xi) ramping to between about 25°C and about
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`40°C or higher for about 15 hours; xii) holding between about 25°C and about 40°C for
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`about 10 hours; xiii) ramping to about 40°C over about 1 hour; and xiv) holding at about
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`40°C for about 5 hours; unloading at about 5°C, at a pressure of about 13.5 psi in a