US008l58607B2
`
`(12) United States Patent
`Singh et al.
`
`(10) Patent N0.:
`(45) Date of Patent:
`
`US 8,158,607 B2
`Apr. 17, 2012
`
`(54) CRYSTALLINE FORM OF LAMIVUDINE
`
`(56)
`
`References Cited
`
`(75)
`
`Inventors: Girij Pal Singh, Pune (IN); Dhananj ai
`Srivastava, Pune (IN); Manmeet
`Brijkishore Saini, Pune (IN); Pritesh
`Rameshbhai Upadhyayt Pune (IN)
`
`U-S- PATENT DOCUMENTS
`5,047,407 A
`9/1991 Belleau
`5,905,082 A
`5/ 1999 R0beITS et all
`EOREIGN PATENT DOCUMENTS
`
`(73) Assignee: Lupin Limited, Mumbai (IN)
`
`$0
`
`(*) Notice:
`
`Subjectto any disclaimer; the term ofthis
`Patent 15 extended or adfllsted under 35
`U-S~C- 15403) by 480 days.
`
`(21) App1.N0.;
`.
`~
`(22) PCT F11ed'
`
`12/297,174
`Feb‘ 9’ 2007
`
`(85) PCT N04
`§ 371 (OX1)
`’
`(2): (4) Date3
`
`PCT/IN2007/000047
`
`0et- 142 2008
`
`(87) PCT Pub. No.: WO2007/119248
`
`PCT Pub. Date: Oct. 25, 2007
`
`(65)
`
`Prior Publication Data
`
`OTHER PUBLICATIONS
`
`(R) M. J. O,Nen et at (eds), “The Merck index, nth Edition,
`” Merck & Co., Whitehouse Station, NJ, 2001, only pp. 958-959
`supplied (see entry “5367,” Lamivudine).*
`Harris, R. K. et al., ‘“‘Polymorphism” in a novel anti-viral agent:
`2653-2659 XP002432450 .
`Lamivudinet,” J. Chem. S(;c., Perkin Trans. 2, vol. 12, 1997, pp.
`Jozwiakowski, M. J., “Solubility Behaviour of Lamivudine Crystal
`Forms in Recrystallisation Solvents,” Journal of Pharmaceutical Sci-
`ences, American Pharmaceutical Association, Washington, US, vol.
`85, No. 2, Feb. 1996, pp. 193-199 (XP-002210585).
`_
`Lachman et al., “The Theory and Practice of Industrial Pharmacy,”
`Third Ed., Varghese Publishing House, Bombay, (1987) pp. 317.
`International Preliminary Report on Patentabililty issued in PCT
`Application No. PCT/IN2007/000047, dated Oct. 21, 2008.
`
`* cited by examiner
`
`Primary Examiner — Lawrence E Crane
`(74) Attorney, Agent, or Firm — Leydig, Voit & Mayer, Ltd.
`
`US 2009/0281053 Al
`
`NOV. 12; 2009
`
`(57)
`
`ABSTRACT
`
`IN ........................... .. 347/KOL/2006
`)
`
`(
`
`(Si)
`
`<30
`A . 18 2006
`pr
`’
`Int CL
`(200000
`A01” 43/50
`(200601)
`A61K 31/50
`(200601)
`A61K 31/70
`(200601)
`C0 7D 239/00
`(200601)
`C07D 239/02
`(52) U.S. Cl.
`......................... .. 514/50; 544/242; 514/247
`(58) Field of Classification Search ...................... .. None
`See application file for complete search history.
`
`f0rm“10‘i00S ‘h0r00f~ A (*9 °iS‘4‘ami00‘1‘(2‘hYdr0XYm‘
`ethyl-1,3-oxathiolan-5-yl)-(1H)-pyrimidin-2-one in the fonn
`iof monoclimc céystlals l:1ai1Ch2tI'aCteI':1S11C1pOV(&ide1I' X-rayi(i111f-
`§Z°c§§sr?§i’ ii aiiiiiieonfiieiliafi .§S°hZff;1a§e3E§aYVéonf.‘
`gosition in Solid dosage unit form compprising a therapeuti-
`cally effective amount of a new Lamivudine polymorphic
`form in combination with a pharmaceutically acceptable car-
`rier is also disclosed along with a pharmaceutical composi-
`tien useful for treating HIV infections in humans.
`
`39 Claims, 16 Drawing Sheets
`
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`(Page 1 of 25)
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`

`
`U.S. Patent
`
`Apr. 17, 2012
`
`Sheet 1 of 16
`
`US 8,158,607 B2
`
`FIG.1
`
`30
`
`
`
`20Position[°2Theta]
`
`10
`
`(\l%
`LO
`’)
`l'
`(N
`
`O<
`
`QC
`
`Counts
`
`1000
`
`500
`
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`

`
`U.S. Patent
`
`Apr. 17, 2012
`
`Sheet 2 of 16
`
`US 8,158,607 B2
`
`[o] €€9'0Z
`
`FIG.2
`
`30
`
`
`
`20Position[°2Theta]
`
`:
`
`10
`
`C°””tS07030610
`
`22500
`
`10000
`
`2500
`
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`

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`U.S. Patent
`
`Apr. 17, 2012
`
`Sheet 3 of 16
`
`US 8,158,607 B2
`
`FIG.3
`
`COUMS20030607
`
`30
`
`
`
`20Position[°2Theta]
`
`“I0
`
`8000
`
`6000
`
`4000
`
`2000
`
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`

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`U.S. Patent
`
`Apr. 17, 2012
`
`Sheet 4 of 16
`
`US 8,158,607 B2
`
`FIG. 4
`
`Delta Y = 1.521%
`
`100.7
`
`100.5
`
`100.0
`
`99.5
`
`99.0
`
`95.5
`
`95.0
`
`
`94.52
`A
`3.578
`
`200
`
`220
`
`240
`
`261.5
`
`20
`
`40
`
`60
`
`80
`
`100
`
`120
`
`140
`
`160
`
`180
`
`Temperature (° C)
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`(Page 5 of 25)
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`

`
`U.S. Patent
`
`Apr. 17, 2012
`
`Sheet 5 of 16
`
`Us 8,158,607 B2
`
`
`
`g
`
`
`
`Temperature (° C)
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`

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`U.S. Patent
`
`Apr. 17, 2012
`
`Sheet 6 of 16
`
`US 8,158,607 B2
`
`FIG. 6
`
`
`
`100.6
`
`100
`
`
`
`
` x1 =so.0o°c
`Y1 =1oo.o1s%
`
`99
`
`X2 =130.00°C
`Y2 = 95.879%
`
`Delta Y = 4.140%
`
`X1 = 140.00°C
`Y1 = 95.829%
`
`0
`—
`-
`X2 = 243.28 C
`Y2 = 93.574%
`
`Delta Y = 2.255%
`
`2.11
`
`20
`
`40
`
`60
`
`80
`
`100
`
`120
`
`140
`
`160
`
`180
`
`200
`
`220
`
`240
`
`254.1
`
`Temperature (° C)
`
`I 98
`
`23
`g 97
`97CD
`5
`
`E.
`
`96
`
`95
`
`94
`
`93.32
`
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`(Page 7 of 25)
`
`

`
`U.S. Patent
`
`Apr. 17, 2012
`
`Sheet 7 of 16
`
`Us 8,158,607 B2
`
`
`
`5.495
`
`5
`
`4 3
`
`2
`
`(3
`
`'2
`
`3
`
`4
`
`'
`
`-5
`
`I
`
`
`
`HeatFlowErrdoUp(mW)
`
`.4‘
`
`A
`
`I
`
`wl
`
`-
`
`'
`
`
`Area = -72.819 mJ
`Delta H = -11.9047 J/g
`
`
`Onset = 127.06°C
`End=131.11°C
`
`
`
`
`‘$008
`60
`so
`100
`120
`140
`150
`130
`200
`220
`240 250-5
`Temperature (°C)
`
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`(Page 8 of 25)
`
`Peak =124.38C
`
`Delta H = 17.9365 J/
`
`Area = 109.415 mJ
`Onset = 11s.27°c
`
`End =
`
`Peak = 177.7o°c
`=
`Q3; Hzfflg
`
`r
`
`Onset = 176.01°C
`End =
`
`

`
`U.S. Patent
`
`Apr. 17, 2012
`
`Sheet 8 of 16
`
`US 8,158,607 B2
`
`66.92
`
`60
`
`50
`
`: 0
`
`
`
`i
`
`_
`O
`Peak — 152.32 c
`Area = 397.887 mJ
`
`e
`
`Delta H = 90.4312 J/g
`
`Onset = 140.86°C
`
`End = 160.O2°C
`
`‘
`
`_
`
`.
`
`:
`
`.
`
`v30
`Q.
`Z)
`O
`‘C
`C
`LIJ
`
`E, 20
`LI.
`
`16CD
`I
`
`
`
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`(Page 9 of 25)
`
`

`
`U.S. Patent
`
`Apr. 17, 2012
`
`Sheet 9 of 16
`
`US 8,158,607 B2
`
`
`
`Peak: 17s.45°c
`Area = 576.239 mJ
`
`; Delta H =112.7919J/g
`
`é
`
`.
`
`:
`
`T
`
`I
`
`,
`
`H
`
`-
`
`16
`
`14
`
`12
`
`10
`
`3
`
`O5
`
`—-mar-u...-.-
`
`4
`
`§g
`
`D
`
`8 2
`C
`LIJ
`
`0
`
`3 E
`
`«(-6
`CD
`
`::_2
`
`4 ‘
`
`
`
`
`
`-6
`
`-8
`
`3.269
`
`20
`
`40
`
`50
`
`80
`
`100
`
`120
`
`140
`
`160
`
`180
`
`200
`
`220
`
`240
`
`261.9
`
`Temperature (°C)
`
`Janssen Ex. 2030
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`(Page 10 of 25)
`
`

`
`U.S. Patent
`
`Apr. 17, 2012
`
`Sheet 10 of 16
`
`US 8,158,607 B2
`
`
`
`108.7
`
`100
`
`80
`
`60
`
`Peak=190.53°C
`
`Area = 648.046 mJ
`
`Delta H = 106.2370 J/g
`
`Onset = 183.41 °C
`
`End = 201 .18°C
`
`*H
`
`I
`I
`
`PHI’
`
`"A '
`
`60
`
`100
`
`120
`
`140
`
`160
`
`180
`
`200
`
`220
`
`244.2
`
`Temperature (°C)
`
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`(Page 11 of 25)
`
`E EQ
`
`402)
`O
`"C
`C
`
`IJJE
`I8
`
`E 20
`
`0
`
`20 '4
`
`-299
`
`49.68
`
`

`
`U.S. Patent
`
`Apr. 17, 2012
`
`Sheet 11 of 16
`
`US 8,158,607 B2
`
`
`
`.
`
`52.99
`
`
`
`*
`
`.
`
`1
`
`Area = 101.333 mJ
`Delta H = 16.6119 J/g
`.
`O
`Onset = 99.72 C
`
`.
`
`End =104.95°C
`
`»
`
`-A
`
`50
`
`48
`
`46
`
`44
`
`A42
`E
`;
`340
`.8
`
`C L
`
`u
`
`Peak = 179.01 °c
`
`Area = 820.259 mJ
`
`Delta H = 101.6818 J/g
`
`Onset = 177.60°C
`End=180.18°C
`
`.
`
`_
`
`.
`
`LI.
`
`-
`E38
`* L
`§36
`‘-3 fiV
`
`‘.
`
`32
`
`1"
`
`
`
`-
`
`30
`29.21
`49.56
`
`60
`
`80
`
`100
`
`120
`
`140
`
`160
`
`180
`
`200
`
`220
`
`240 250.8
`
`Temperature (°C)
`
`Janssen Ex. 2030
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`Lupin Ltd. v. Janssen Sciences Ireland UC
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`(Page 12 of 25)
`
`

`
`U.S. Patent
`
`Apr. 17, 2012
`
`Sheet 12 of 16
`
`US 8,158,607 B2
`
`164.7
`160
`
`140
`
`
`
`120
`
`I 100
`
`-
`
`Peak = 123.oo°c
`
`Area = 690.621 mJ
`Delta H = 113.2166 J/g
`
`Onset = 115.28°C
`0
`
`End=130.71 c
`
`Peak = 151 .70°C
`Area = 33.473 mJ
`Delta H = 5.4874.J/g
`
`Onset = 141 .62°C
`
`_
`
`A
`
`_
`
`'
`
`.
`
`I
`
`1
`
`.
`
`L‘
`
`3
`
`EE
`
`Q.
`:)
`
`'§ 50
`
`so
`
`L;
`E
`*5
`G, 40I
`
`
`
`-15.14
`
`
`3
`
`51.83
`
`60
`
`so
`
`100
`
`120
`
`140
`
`160
`
`180
`
`200
`
`220
`
`2453
`
`Temperature (°C)
`
`Janssen Ex. 2030
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`Lupin Ltd. v. Janssen Sciences Ireland UC
`|PR2015-01030
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`(Page 13 of 25)
`
`

`
`U.S. Patent
`
`Apr. 17, 2012
`
`Sheet 13 of 16
`
`US 8,158,607 B2
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`Lupin Ltd. v. Janssen Sciences Ireland UC
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`(Page 14 of 25)
`
`

`
`U.S. Patent
`
`Apr. 17, 2012
`
`Sheet 14 of 16
`
`US 8,158,607 B2
`
`8006000
`
`1000
`
`1200
`
`1400
`28002400200018001600
`
`
`
`
`cm
`
`FIG.14
`
`“‘~|
`
`534
`
`3200
`
`
`
`400003600
`
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`(Page 15 of 25)
`
`

`
`U.S. Patent
`
`Apr. 17, 2012
`
`Sheet 15 of 16
`
`US 8,158,607 B2
`
`FIG.15
`
`600.0
`
`800
`
`
`
`1200I1000
`
`
`
`16002‘1400
`
`20001800
`
`2400
`
`2800
`
`3200
`
`32600
`
`'4ooo.o
`
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`(Page 16 of 25)
`
`

`
`U S. Patent
`
`Apr. 17, 2012
`
`Sheet 16 of 16
`
`US 8,158,607 B
`
`FIG. 16
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`

`
`US 8,158,607 B2
`
`2
`
`1
`CRYSTALLINE FORM OF LAMIVUDINE
`
`CROSS-REFERENCE TO RELATED
`APPLICATIONS
`
`This patent application is a U.S. National Phase of Inter-
`national Patent Application No. PCT/IN2007/000047, filed
`Feb. 9, 2007, which claims the benefit of India Patent Appli-
`cation No. 347/KOL/2006, filed Apr. 18, 2006, each of which
`are incorporated herein by reference i11 their entireties.
`
`FIELD OF INVENTION
`
`The present invention relates to a new Lamivudine poly-
`morphic form, pharmaceutical formulations thereof.
`
`BACKGROUND OF THE INVENTION
`
`Lamivudine (I) (CAS No. 134678-17-4) is chemically
`known as (2R-cis)-4-arnino-1-[2-(hydroxymethyl)-1,3 -ox-
`athiolan-5 -yl]-2(1H)-pyrimidinone, also known as (—) cis-4-
`amino-1-(2-hydroxymcthyl-1,3-oxathiolan-5 -yl)-(1H)-pyri-
`midin-2-one
`
`(1)
`
`NH2
`
`N \
`
`/
`N
`
`0
`
`s
`
`HO O
`
`R
`
`S
`
`Lamivudine is a reverse transcriptase inhibitor used in the
`treatment of HIV infection alone or in combination with other
`
`class ofAnti HIV drugs.
`Lamivudine is commercially available in a pharmaceutical
`composition under the brand name EPIVIR® marketed by
`GlaxoSmithKine and is covered under U.S. Pat. No. 5,047,
`407.
`U.S. Pat. No. 5,047,407 claims 1,3-oxathiolane deriva-
`tives,
`their geometric and optical
`isomers and mixtures
`thereof. The patent also discloses the preparation of cis and
`trans isomers of 2,5 substituted 1,3-oxathiolane derivatives.
`U.S. Pat. No. 5,905,082 describes two polymorphic modi-
`fications of Lamivudine viz form I and II. Form 1 crystals are
`short rods or long thin needles with orthorhombic crystal
`system. Form 1 is a hydrate of Lamivudine consisting of one
`molecule of water per five molecules of Lamivudine. This
`form melts at 146° C. (Journal ofChem. Soc., Perkin Trans. 2,
`page 2655 (1997)). The DSC thermogram (the rate ofheating:
`2° C./min) of this form shows first an endotherm at 123.6° C.
`followed by an exotherm at 128° C., finally another endot-
`herm at 179.6° C. This second endotherm is due to conversion
`
`of crystal form I to form II, hence form 1 is a metastable
`crystalline form.
`However with rate of heating of 100° C./min form I shows
`a single endotherm at 146° C., which is it’s melting point. The
`TGA shows a single step sharp weight loss of 2%.
`Form I as per U.S. Pat. No. 5,905,082 is prepared by
`heating a suspension of 64.8 gm Lamivudine in 200 ml water
`at 45° C. to give a solution and cooling the solution to 30° C.
`
`10
`
`15
`
`20
`
`25
`
`The product crystallizes out as an unstirrable mass. Further
`breaking this mass and cooling it to 10° C. with stirring and
`thereafter filtering and drying at 45° C. for 24 hours gives
`form I crystals.
`Form II crystals as disclosed in U.S. Pat. No. 5,905,082 are
`bipyramidal in shape with tetragonal crystal system. It is an
`anhydrous form of Lamivudine. This form melts at 177° C.
`(Journal of Chem. Soc., Perkin Trans. 2, page 2655 (1997)).
`The DSC thermogram of this form at all scan speeds shows a
`single peak of endotherm at 177° C. Form II is a stable
`crystalline form of Lamivudine and is claimed in U.S. Pat.
`No. 5,905,082.
`Form II as per U.S. Pat. No. 5,905,082 is prepared by
`following procedure: Heat a suspension of 10 gm Lamivudine
`in 200 ml of industrial methylated spirit to reflux to obtain a
`clear solution. Filter the solution while hot; distil half the
`amount of the solvent from the filtrate then stop heating and
`seed the concentrated solution with authentic form II crystals.
`The seeded solution is then cooled from 80° C. to 25° C.
`during one hour. Crystal formation starts at 79° C. Further
`cooling the suspension to 15° C. and stirring for an hour,
`filtration, washing with IMS and drying gives Form II crys-
`tals.
`Crystalline form I have inferior flow property and also
`lower bulk density, which create problem in handling the
`product during formulation. In view of the literature cited
`hereinbefore Lamivudine form I also suffers from stability
`issues. Therefore, it is desirable to develop a crystalline fonn
`of Lamivudine having improved stability and also compa-
`rable if not better bioavailability.
`30 When slurried in water both crystal form I and II get con-
`verted to another polymorphic form not yet reported in the
`literature, which is really not a desirable feature for manufac-
`turing practices. Form I converts to form II during milling and
`formulation operation and because of this the invention
`embodied in U.S. Pat. No. 5,905,082 for getting form II, a
`35 thermodynamically stable polymorph, used for formulation.
`The present inventors have surprisingly i found that Larni-
`vudine can also be obtained in a third crystalline form (here-
`inafter form III), which not only have distinct powder X-ray
`diffractogram but also have entirely different single crystal
`X-ray diffraction when compared to form I and II.
`
`40
`
`OBJECTS OF THE INVENTION
`
`Thus an object ofthe present invention is to provide a novel
`crystalline hemihydrate form of Lamivudine with better flow
`property and bulk density, which enables to have a formula-
`tion without any difiiculty.
`Another object of the present invention is to provide a
`novel crystalline hemihydrate form of Lamivudine with com-
`parable dissolution rate with the reported polymorphic forms
`of larnivudine.
`Yet another object of the present invention is to provide a
`novel crystalline form ofLamivudine that is stable during wet
`granulation using water as a granulating solvent, thereby
`ensuring the physical stability of the finished solid dosage
`form.
`
`A further object of the present invention is to provide a
`process for preparation of novel crystalline hemihydrate of
`Lamivudine using eco-friendly solvent “water”.
`Another object of the present invention is to provide suit-
`able pharmaceutical dosage forms of novel crystalline hemi-
`hydrate of Lamivudine alone or in combination with other
`anti HIV agents.
`
`SUMMARY OF INVENTION
`
`45
`
`50
`
`55
`
`60
`
`65
`
`Thus in the present invention there is provided a crystalline
`hemihydrate (form III) of Lamivudine having characteristic
`
`Janssen Ex. 2030
`
`Lupin Ltd. v. Janssen Sciences Ireland UC
`|PR2015-01030
`
`(Page 18 of 25)
`
`

`
`US 8,158,607 B2
`
`3
`powder and single crystal X-ray diffraction as shown in FIGS.
`1 and 16 with characteristic 20 values as given in Table III.
`According to another aspect of the present invention there
`is provided a method for formation of Fonn III by dissolving
`Lan1ivudine in water at 45° C., then cooling the clear solution 5
`to 30° C., optionally seeding with form III crystals and further
`cooling to 10° C. at the rate ranging from 0.5° C./min to 3.5°
`C./min, isolating the crystals by filtration optionally washing
`with alcohol and drying at 45-55° C.
`
`10
`
`DESCRIPTION OF THE INVENTION
`
`As mentioned earlier both form I and form II polymorphs
`when slurried in water get converted to polymorphic form III,
`which happens to be thermodynamically stable and does not
`undergo any change in crystal structure during milling.
`This crystal form has been found to have better flow prop-
`erty and higher bulk density in comparison with literature
`reported forms.
`Further study on single crystal X-ray diffraction reveals
`that it is a hemihydrate form (four molecules of Lamivudine
`with two molecules of water) of Lamivudine. This product
`melts at 176-177° C. The DSC thermogram (at the rate of
`
`15
`
`20
`
`TABLE I
`
`Form I
`
`0.46
`0.60
`33.66°
`
`Form II
`
`Form III
`
`0.38
`0.55
`32.00°
`
`0.64
`0.83
`32.00°
`
`Property
`
`Bulk Density (gm/cc)
`Tap Density (gm/cc)
`Flow Property
`(Angle ofRepose$)
`
`$measured as per the procedure provided on page 317 of ‘The Theory and Practice of
`Industrial Pharmacy’ by Leon Lachman et al., Third Ed. Varghese Publishing House, Bom-
`bay; (1987)
`
`Lamivudine Form I and Form II when slurried in water at
`
`ambient temperature for 24 to 48 hours get converted to Fonn
`III, which is not at all desirable since during formulation
`especially in wet granulation such conversion would lead to
`physical instability of the finished formulation. Hence, use of
`Lamivudine Form III crystals would certainly have an added
`advantage over other polymorphic forms mentioned in the
`literature.
`
`The crystalline form III of Lamivudine as disclosed herein
`was found to be stable for more than three months when
`stored at 40:2° C. RH 75:5%.
`
`Comparative thermal analysis data is tabulated in Table II
`TABLE II
`
`Crystal
`Form
`
`Melting Point
`
`DSC
`
`I
`
`II
`
`135-145° C.
`124-127° C.*
`135° C.#
`
`@ 2° C./min: exotherm at 123°
`then at 177° (FIG. 7)
`@ 100° C./min: 146° C. (FIG. 8)
`
`177-178° C.
`177—178° c.*#
`
`@ 2° C./min and 100° C./min:
`177° C. (FIG. 9 & 10)
`
`III
`
`176-177° C.
`
`@ 2° C./min first peak at 100° C.
`and second at 177° C. (FIG. 11)
`@100° C./min: 120° C. (FIG. 12)
`
`TGA
`
`One step weight loss
`between temp 80° C. to
`140° C. = 1.52%
`(FIG. 4)
`No weight loss due to
`crystal bound water.
`(FIG. 5)
`One step weight loss
`between temp 80° C. to
`140° C. = 4.14%
`(FIG. 6)
`
`heating:2° C./min) shows first peak of endotherm (A
`H:l 6.61 J/g) at 100° C. and the second peak ofendotherm (A
`H:l01.68 J/g) at 179.6°. This crystal form is found to be
`stable and has better flow property than form 1, and is found
`to posses comparable bioavailability.
`The crystal form III of Lamivudine is obtained by subject-
`ing the hot (45° C.) supersaturated solution of Lamivudine for
`controlled cooling. Whereas if such solution is cooled sud-
`denly it gives form 1 crystals of Lamivudine.
`
`40
`
`45
`
`50
`
`Therrnogravimetric analysis (as shown in FIG. 6) of form
`III crystals of Lamivudine shows 3.5 to 4% single step loss of
`weight. Moisture content of this crystal form by Karl Fischer
`titration is in the range of 3.5 to 4.0%, which confirms pres-
`ence of approximately one mole ofwater per every two moles 5
`of Lamivudine.
`
`5
`
`Single crystal structure X-ray data (FIG. 16) reveals two
`molecules of water are associated with four molecules of
`lamivudine presumably through hydrogen bonds in polymor-
`phic forrn III. In other words the material ofpresent invention
`is a hemihydrate having four molecules of lamivudine and
`two molecules of water. Form III thus obtained has a melting
`point of 176 to 177° C.
`The novel crystalline hemihydrate form (form III) ofLarni-
`vudine has better flow property and bulk density, which are
`important parameters for formulation (Table I).
`
`60
`
`65
`
`The powder X-ray diffraction analysis of form III also
`shows characteristic 20 values. Comparative data of 20 values
`form III and other literature reported polymorphic forms is
`provided in Table III
`
`Form I (FIG. 1)
`(20 values)
`5.20
`6.66
`8.53
`8.81
`9.65
`9.85
`10.15
`10.41
`11.27
`11.38
`11.63
`12.34
`12.60
`12.93
`13.22
`14.60
`15.01
`15.17
`15.67
`15.81
`
`TABLE III
`
`Form II (FIG. 2)
`(20 values)
`10.70
`12.17
`13.42
`14.30
`14.76
`15.86
`16.83
`17.55
`18.63
`19.68
`20.63
`21.44
`22.13
`22.60
`23.03
`24.44
`24.94
`25.70
`26.51
`27.68
`
`Form III (FIG. 3)
`(20 values)
`5.50
`7.60
`9.00
`9.62
`0.98
`1.97
`2.52
`2.81
`3.52
`5.19
`5.71
`5.94
`6.57
`6.72
`7.11
`7.57
`7.98
`8.30
`9.26
`9.68
`
`Janssen Ex. 2030
`
`Lupin Ltd. v. Janssen Sciences Ireland UC
`|PR2015-01030
`
`(Page 19 of 25)
`
`

`
`US 8,158,607 B2
`
`5
`TABLE III-continued
`
`Form I (FIG. 1)
`(26 values)
`
`Form II (FIG. 2)
`(26 values)
`
`Form III (FIG. 3)
`(26 values)
`
`16.51
`17.59
`
`17-93
`18.13
`18.72
`1910
`19.30
`19-75
`3471::
`23706
`25.44
`3-901
`'
`29.46
`31_00
`
`28.41
`28.93
`
`29-72
`30.67
`30.90
`3130
`31.47
`31-99
`37513
`33_14
`34.01
`3213
`'
`37.27
`3g_46
`
`20.37
`21.04
`
`22-00
`22.86
`23.40
`2370
`24.04
`24-58
`
`27_70
`28.74
`38-23
`'
`31.94
`3325
`
`The single crystal X-ray diffraction data obtained for form
`III crystalline form of Lamivudine is tabulated in Table IV
`Suitable pharmaceutical formulations may conveniently
`be presented containing predetermined amount oflamivudine
`in crystalline form III
`
`DESCRIPTION OF ACCOMPANYING FIGURES
`
`FIG. 1: Powder X-ray diffractogram ofcrystalline form I of
`Lamivudine.
`
`FIG. 2: Powder X-ray diffractogram of crystalline form II
`of Lamivudine.
`
`FIG. 3: Powder X-ray diffractogram of crystalline form III
`of Lamivudine.
`
`FIG. 4: TGA thermogram of crystalline form I of Lamivu-
`dine.
`
`FIG. 5: TGAthermogra1n of crystalline form II of Lami-
`vudine.
`
`6
`The present invention is illustrated in more detail by refer-
`ring to the following Examples, which are not to be construed
`as limiting the scope of the invention.
`
`EXAMPLE 1
`
`Preparation of Lamivudine Form III
`,
`,
`,
`,
`A suspension of the Lamivudine forrn-II (25.0) g in water
`(75.0 ml) was heated to 45° C. in 20 min to give a clear
`solution. The solution was cooled to 30° C. during a period of
`30 min. The crystallization. started at 30° C. The masswas
`further cooled to 10° C. during a period of20 min and stirred
`for 1 hour. The product was filtered and washed with ethanol
`(2><l0 ml) then dried in vacuum at 45° C. for 24 hours.
`Yield:23.0 gms.
`,
`_
`Spectra [Nu]Ol
`(CII1 1)Z
`1640,1600,1522,1460,1376,1296,1226,1193,1155,1135,
`1106, 1044, 976, 927, 844, 788, 722 (FIG. 15)
`X-ray powder diffraction analysis shows peaks at about
`5.50, 7.60, 9.00, 9.62, 10.98, 11.97, 12.52, 12.81, 13.52,
`15.19,15.71,15.94,16.57,16.72,17.11,17.57,17.98,18.30,
`19.26, 19.68, 20.37, 21.04, 22.00, 22.86, 23.40, 23.70, 24.04,
`24.68, 25.15, 26.97, 27.70, 28.74, 30.35, 30.60, 31.94,
`33.25:0.2°26.
`
`The single crystal X-ray analysis is carried out using
`SMART APEX CCD diffractometer by full-matrix least-
`squares refinement on F2; goodness of fit on F2 was 1.050. A
`total of 20474 reflections were measured on diffractometer
`with monochromatised Cu—K0t radiation. The data was col-
`
`lected at 6 ranging from 1.26 to 25°. The structure was solved
`by direct method and the non-hydrogen atoms refined aniso-
`tropically. All H atoms were refined isotropically. Refinement
`converged to give R1:0.0538, wR2:0.1428. Minimum
`residual electron density was -0.403 e. A‘3 and maximum
`residual electron density was 0.887 A‘3 . The data is as shown
`below in Table IV:
`
`5
`
`10
`
`15
`
`20
`
`25
`
`30
`
`35
`
`FIG. 6: TGA thermogram of crystalline form III of Lami- 40
`.
`Vud1ne'
`FIG. 7: DSC thermogram of crystalline form I of Lamivu-
`dine at heating rate 20 C'/min'
`FIG. 8: DSC thermogram of crystalline form I ofLamivu- 45
`dine at heating rate 100° C./min.
`
`FIG. 9: DSC thermogram of crystalline form II of Lami-
`vudine at heating rate 2° C./min.
`FIG. 10: DSC thermogram of crystalline form II of Lami-
`vudine at heating rate 100° C./min.
`FIG. 11: DSC thermogram of crystalline form III of Lami-
`vudine at heating rate 2° C./min.
`FIG. 12: DSC thermogram of crystalline form III of Lami-
`vudine at heating rate 100° C./min.
`FIG. 13: FTIR spectra of crystalline form I of Lamivudine.
`FIG. 14: FTIR spectra of crystalline form II ofLamivudine.
`FIG. 15: FTIR spectra of crystalline form III of Lamivu-
`dine.
`
`FIG. 16: crystal structure and packing diagram of crystal-
`line form III of Lamivudine obtained by Single crystal X-ray
`diffraction analysis. (a) Crystal structure of Lamivudine form
`III. (Disordered atom of minor component S9D1 has been
`omitted for clarity.) (b) Packing diagram of Lamivudine form
`III. (Disordered atom of minor component S9Dl has been
`omitted for clarity.)
`
`50
`
`55
`
`60
`
`65
`
`,
`,
`Empirical Formula
`Formula weight
`CYYS131 System
`§I1if1tc:§llOd1fnensions
`
`Z, calculated density
`Cellvolume
`C1YS1a1S1Z€
`
`TABLE IV
`
`2(C8H11N3O3S)°(H2O)
`476.53
`1V1°H°°11H1°
`§2=l11.714 (9) A
`b = 11-214 (9)21
`° = 16197 (1210
`2, 1.493 Mg/m3.
`2120.4 (3)213
`0-18 X 0-11 X 0-09
`
`OL = 90°
`[5 = 94-63°
`7 = 990
`
`Powder pattern generated from single crystal data using
`MERCURY software was found to be identical to the experi-
`mental powder X-ray diffraction pattern of the material of
`invention (as provided for Form III in Table III and in FIG. 3).
`The differential scanning calorimetric analysis at the rate
`ofheating 2° C./min shows first peak of endotherin at 100° C.
`and second at 177° C. (FIG. 11), and at the rate of heating
`100° C./min shows single peak of endotherm at 120° C. (FIG.
`12).
`The thermogravimetric analysis exhibits one-step weight
`loss of4.14% between temp 80° C. to 140° C. (FIG. 6).
`
`EXAMPLE 2
`
`Preparation of Lamivudine Form III
`
`A suspension of the Lamivudine forrn-II (20.0) g in water
`(60.0 ml) was heated to 45° C. in 25 min to give a solution.
`
`Janssen Ex. 2030
`
`Lupin Ltd. v. Janssen Sciences Ireland UC
`|PR2015-01030
`
`(Page 20 of 25)
`
`

`
`US 8,158,607 B2
`
`7
`The solution was cooled to 30° C. in 15 min. The mass was
`
`then cooled to 10° C. in 20 min and stirred for 1 h. The product
`was filtered and washed with IMS (2><10 ml) then dried in
`vacuum at 45° C. for 24 h. Yield:17 gms.
`Powder X-ray diffraction pattern superimposable with that
`of form 111 as obtained in Example 1.
`
`8
`Powder X-ray diffraction pattern superimposable with that
`of form 111 as obtained in Example 1.
`
`EXAMPLE 8
`
`Preparation of Lamivudine Form 111
`
`EXAMPLE 3
`
`Prcparation of Lamivudinc Form 111
`
`A suspension of the Lamivudine form-ll (20.0) g in water
`(60.0 ml) was heated to 45° C. in 25 min to give a solution.
`The solution was cooled to 30° C. in 30 min. The mass was
`
`then cooled to 10° C. in 20 min and stirred for 1 h. The product
`was filtered and washed with ethanol (2><l0 ml), then dried in
`vacuum at 45° C. for 24 h. Yield:17 gms.
`Powder X-ray diffraction pattern superimposable with that
`of form 111 as obtained in Example 1.
`
`EXAMPLE 4
`
`Preparation of Lamivudine Form 111
`
`A suspension of the Lamivudine form-ll (10.0) g in water
`(30.0 ml) was heated to 45° C. in 20 min to give a clear
`solution. The solution was cooled to 30° C. in 15 min. The
`reaction mass was then cooled to 10° C. in 20 min and stirred
`for 1 h. The product was filtered and dried in vacuum at 45° C.
`for 24 h. Yield:8.5 gms.
`Powder X-ray diffraction pattern superimposable with that
`of form 111 as obtained in Example 1.
`
`EXAMPLE 5
`
`Preparation of Lamivudine Form 111
`
`A suspension of the Lamivudine form-1 (10.0) g in water
`(30.0 ml) was heated to 45° C. in 20 min to give a clear
`solution. The solution was then cooled to 10° C. in 10 min and
`stirred for 1 11. Tl1e product was filtered and dried ir1 vacuur11
`at 45° C. for 24 h. Yield:7 gms
`Powder X-ray diffraction pattern superimposable with that
`of form 111 as obtained in Example 1.
`
`EXAMPLE 6
`
`Preparation of Lamivudine Form 111
`
`10
`
`15
`
`20
`
`25
`
`30
`
`35
`
`40
`
`45
`
`A suspension of the Lamivudine form-H (20.0) g in water
`(80.0 ml) was heated to 45° C. in 25 min to give a clear
`solution. Thc solution was coolcd slowly to 30° C. in 55 min.
`The product was then cooled to 10° C. in 5 min and stirred for
`1 h at the same temperature. The product was filtered and
`dried in vacuum for 24 hr at 50-55° C. Yield: 18 gm.
`Powder X-ray diffraction pattern superimposable with that
`of form 111 as obtained in Example 1.
`
`EXAMPLE 9
`
`Preparation of Lamivudine Form 111
`
`A suspension of the Lamivudine form-H (20.0) g in water
`(100.00) was heated to 45° C. in 25 min to give a clear
`solution. The solution was cooled slowly to 30° C. in 55 min.
`The product was then cooled to 10° C. in 5 min and stirred for
`1 h at the same temperature. The product was filtered and
`dried in vacuum for 24 hr at 50-55° C. Yield 18.7 gm.
`Powder X-ray diffraction pattern superimposable with that
`of form 111 as obtained in Example 1.
`
`EXAMPLE 10
`
`Preparation of Lamivudine Form 111
`
`A suspension of lamivudine (Form 1 or Form 11 or mixture
`thereof) (35 gm) in water (105 ml) was heated to 45° C. in 17
`minutes to give a clear solution. The solution was cooled
`slowly to 37° C. in 50 minutes. The solution was seeded with
`lamivudine form 111. The mixture was then cooled to 10° C. in
`
`10 minutes and stirred for one hour. The product was filtered
`and dried ir1 vacuum at 45° C. for 24 hours. Yield 32 gr11.
`Powder X-ray diffraction pattern superimposable with that
`of form 111 as obtained in Example 1.
`
`EXAMPLE 11
`
`Preparation of Lamivudine Form 111
`
`A suspension of the Lamivudine form-ll (5.0 gm) in water
`A suspension of the Lamivudine form-ll (10.0) g in water 50 (5.0 ml) was stirred at 25° C. for 48 hours. The suspension
`(30.0 ml) was heated to 45° C. in 20 min to give a clear
`was cooled and stirred at 10° C. for one hour. The product was
`solution. The solution was then cooled to 10° C. in 10 min and
`filtered and then dried under vacuum at 45° C. for 24 hours.
`
`stirred for 1 hr. The product was filtered and dried in vacuum
`at 45° C. for 24 hr. Yield:8 gm.
`Powder X-ray diffraction pattern superimposable with that
`of form 111 as obtained in Example 1.
`
`EXAMPLE 7
`
`Preparation of Lamivudine Form 111
`
`A suspension of the Lamivudine form-ll (50.0) g in water
`(150.0 ml) was heated to 45° C. in 17 min. to give a clear
`solution. The solution was cooled slowly to 30° C. in 1.0 hr 40
`min. The product was then cooled to 10° C. in 10 min and
`stirred for 1 h. The product was filtered and dried in vacuum
`1.0 mm at 45° C. for 24 h. Yield:44 gm
`
`55
`
`60
`
`65
`
`Yield:4.5 gms
`Powder X-ray diffraction pattern superimposable with that
`of form 111 as obtained in Example 1.
`
`EXAMPLE 12
`
`Preparation of Lamivudine Form 111
`
`A suspension of the Lamivudine form-I (2.0 gm) in water
`(2.0 ml) was stirred at 25° C. for 24 hours. The suspension
`was cooled and stirred at 10° C. for one hour. The product was
`filtered and then dried under vacuum at 45° C. for 24 hours.
`
`Yield:1.6 gms
`Powder X-ray diffraction pattern superimposable with that
`of form 111 as obtained in Example 1.
`
`Janssen Ex. 2030
`
`Lupin Ltd. v. Janssen Sciences Ireland UC
`|PR2015-01030
`
`(Page 21 of 25)
`
`

`
`9
`EXAMPLE 13
`
`Preparation of Lamivudine Form I
`
`US 8,158,607 B2
`
`10
`The thermogravimetric analysis reveals that it is an anhy-
`drous product. (FIG. 5).
`
`EXAMPLE 12
`
`Pharmaceutical Formulations
`
`(a) 150 mg Lamivudine Tablet
`
`Ingredients per Tablets
`
`Weight (mg.)
`
`(_F°““ 111)
`L3_miVudiH€
`§’i1§fjI;1§f:jf£“;1;:flj:j:fi51‘;F
`co11o1da1s111con dioxidg N1:
`Magnessium stearateNF
`
`Totalweight
`
`150
`
`2_25
`5.63
`
`45000
`
`Lamivudine (form III), microcrystalline cellulose, sodium
`starch glycolate and colloidal silicon dioxide were sieved and
`blended in octagonal for about 15 minutes. Sieved magne-
`sium stearate was then added and blending continued for a
`further 2 minutes
`
`The blend was compressed in standard tabletting equip-
`ment.
`
`Analysis:
`Tablet weight: 450 mg:5%
`Thickness: 5.0-5.2 mm
`Hardness: 150 to 200 N
`
`Disintegration Time: 25 seconds.
`% friability: 0.1%.
`(b) Larnivudine Form III/Zidovudine Combination Tablets:
`
`EXAMPLE”
`.
`.
`.
`Preparation of Lamivudine Form II
`
`E;Zi‘Li;”:?£f§2?;2I§1i:§§3i;“°NF
`P 'fi d
`t
`E::rt:a.egr:n:1e;:r
`
`45
`
`A suspension ofthe Lamivudine (10.0) g in water (30.0 ml)
`was heated to 45° C. in 30 min to give a solution. The solution
`was cooled to 30° C. in 0.5 min. The product was crystallized
`as an unstirrable mass. This was broken up and suspension
`stirred at 10.0° C. for 1 hr. The product was filtered and
`washed with IMS (2><5 ml) then dried in vacuum at 45° C. for 10
`24 hi Yie1d:6'0 gm
`IR Spectra [Nujol Mull] (cm‘1): 3356, 3199, 2923, 2854,
`1639,1611,1461,1402,1376,1309,1288,1252,1196,1166,
`1145, 1107, 1052, 970, 932, 839,786,720 (FIG. 13).
`X-ray powder diffraction analysis shows peaks at about
`5.20, 6.66, 8.53, 8.81, 9.65, 9.85, 10.15, 10.41, 11.27, 11.38,
`
`15
`
`11.63, 12.34,12.60, 12.93,13.22,14.60,15.01,15.17, 15.67,
`15.81, 16.51,17.59, 17.98,18.13,18.72,19.10,19.30, 19.76,
`21.79,
`23.49,
`23.71,
`25.44,
`25.90,
`27.34,
`29.46, 20
`31 .00:0.2°20.
`
`The differential scarming calorimetric analysis at the rate
`ofheating 2° C./min shows first peak of endotherm at 123° C.
`and second at 177° C. (FIG. 7), and at the rate ofheating 100°
`C./min shows single peak of endotherm at 146° C. (FIG. 8).
`The thermogravimetric analysis exhibits one-step weight
`loss of 1.52% between temp 80° C. to 140° C. (FIG. 4).
`
`EXAMPLE 14
`
`Preparation of Lamivudine Form I
`
`25
`
`30
`
`A suspension ofthe Lamivudine (250.0 g) in the mixture of
`water (750.0 ml) and DNS (250.0 ml) was heated to 45° C. in
`12 min to give a solution. The solution was cooled to 30° C. in
`15 min and seeded with form I crystals. The product was then
`cooled to 10° C. in 30 min and stirred for 1 h. The product was
`filtered washed wished with 100 ml water DNS mixture (3:1)
`.
`.
`.
`.
`and dried in vacuum at 45° C. for 24 h. Yield: 220.0 gm.
`Powder X-ray diffraction pattern supenmposable with that 40
`of form I as obtained in Example 13.
`
`35
`
`A suspension ofthe I,amivudine (10.0) g in ethanol (200.0
`.
`.
`ml) was heated to refluxed to give a clear solution. The solu-
`tion thus formed was subjected to distillation and about 100
`ml of ethanol was distilled out at atmospheric pressure. The 50
`remaining solution was then cooled to 15° C. in 35 min. The
`suspension stirred at 15° C. for 1.0 hr. The product was
`filtered and washed with ethanol (10.0 ml) then dried in
`vacuum at 50° C. for 12 hr to get 8.2 gm.
`IR Spectra [Nujol Mun] (Cm—1): 3322, 3194, 2950, 2870, 55
`1651,1611,1496,1