1111111111111111 IIIIII IIIII 11111 1111111111 lllll 111111111111111 11111 11111 1111111111 11111111
`US 20060247189Al
`
`c19) United States
`c12) Patent Application Publication
`Ionescu et al.
`
`c10) Pub. No.: US 2006/0247189 Al
`Nov. 2, 2006
`(43) Pub. Date:
`
`(54) FORMS OF 5-AZACYTIDINE
`
`Publication Classification
`
`(75)
`
`Inventors: Dumitru Ionescu, Ann Arbor, MI (US);
`Peter Blumbergs, Royal Oak, MI (US);
`Gary L. Silvey, Overland Park, KS
`(US)
`
`(51)
`
`Int. Cl.
`C07H 191048
`(2006.01)
`A61K 3117072
`(2006.01)
`(52) U.S. Cl. ............................................. 514/43; 536/28.1
`
`Correspondence Address:
`SWANSON & BRATSCHUN L.L.C.
`1745 SHEA CENTER DRIVE
`SUITE 330
`HIGHLANDS RANCH, CO 80129 (US)
`
`(57)
`
`ABSTRACT
`
`The invention provides novel polymorphic and pseudopoly(cid:173)
`morphic crystalline forms of 5-azacytidine, along with
`methods for preparing said forms, wherein 5-azacytidine is
`represented by the formula:
`
`(73) Assignee: Pharmion Corporation, Westminster,
`CO (US)
`
`(21) Appl. No.:
`
`11/458,365
`
`(22) Filed:
`
`Jul. 18, 2006
`
`Related U.S. Application Data
`
`(60) Continuation of application No. 11/052,615, filed on
`Feb. 7, 2005, now Pat. No. 7,078,518, which is a
`division of application No. 10/390,578, filed on Mar.
`17, 2003, now Pat. No. 6,887,855.
`
`HO
`
`0
`
`OH
`
`OH
`
`The invention also includes pharmaceutical compositions
`comprising said forms.
`
`CELGENE 2038
`APOTEX v. CELGENE
`IPR2023-00512
`
`

`

`Patent Application Publication Nov. 2, 2006 Sheet 1 of 8
`
`US 2006/0247189 Al
`
`Figure 1.
`
`X-ray Powder Diffraction Pattern of Azaciti.dine, Form I, Labeled with the
`more Prominent 28 Angles (Cu Ka Radiation)
`
`5500
`
`5000
`
`4500
`
`4000
`
`3500
`
`3000
`
`,t, ·- 2500
`!
`.!! = 2000
`1500
`
`~
`
`1000
`
`500
`
`0
`
`5
`
`10
`
`15
`
`20
`
`25
`
`30
`
`35
`
`40
`
`Two-Theta
`
`

`

`Patent Application Publication Nov. 2, 2006 Sheet 2 of 8
`
`US 2006/0247189 Al
`
`Figure 2 X-ray Powder Diffraction Pattern of A7.acitidine, Mixed Phase Forms I and II,
`Labeled with the more Prominent 29 Angles (Cu Ka Radiation)
`
`• indicates fines distinctive to Form II
`
`b
`Ei
`~
`
`1-,
`i!S
`,..;
`~-~
`,-,i ~
`
`- ~ ~
`~
`
`0
`
`0
`
`~ §
`
`-
`
`N
`
`00
`
`.,,
`~
`
`~
`!!i
`
`....
`~
`
`00
`
`*
`~
`el
`~ i::i
`1-,
`i;;;
`i::i
`
`~
`
`a
`
`5000
`
`4500
`
`4000
`
`3500
`
`3000
`
`·-
`t;, 2500
`~
`-
`i 2000
`1500
`
`1000
`
`Azacitidine
`500 Mixed Form; I & II
`
`0
`
`5
`
`JO
`
`15
`
`20
`
`25
`
`30
`
`35
`
`40
`
`Two-Theta
`
`

`

`Patent Application Publication Nov. 2, 2006 Sheet 3 of 8
`
`US 2006/0247189 Al
`
`Figure 3 X-ray Powder Diffraction Pattern of A7Jlcitidine, Form III, Labeled with the
`more Prominent 28 Angles (Cu Ka Radiation)
`
`4000
`
`3500
`
`3000
`
`2500
`
`2000
`
`...
`.0
`ltl
`-
`=
`CII 1500
`
`1000
`
`500
`
`0
`
`:0
`~
`
`~
`
`~
`:ti
`
`M
`~
`
`~
`!:::
`
`00
`~
`'!i
`
`el
`"
`-'
`N
`
`g
`"'
`
`""!
`
`~
`
`~
`~
`
`:2
`~
`
`~
`~
`~ ::i
`
`~
`~
`
`=::
`i;i
`
`~
`;;
`
`5
`
`10
`
`15
`
`20
`
`25
`
`30
`
`35
`
`40
`
`Two-Theta
`
`

`

`Patent Application Publication Nov. 2, 2006 Sheet 4 of 8
`
`US 2006/0247189 Al
`
`Figure 4 X-ray Powder Diffraction Pattern of Azacitidine, Form IV, Labeled with the
`more Prominent 20 Angles (Cu Ka Radiation)
`
`3000
`
`2500
`
`2000
`
`1500
`
`1000
`
`500
`
`0
`
`.e-
`....
`~
`-
`i
`
`...
`.,..
`;:j
`
`b
`E=i
`;:!;
`
`~
`ct
`'.d
`
`~
`:e
`
`~
`...,
`.,..
`
`~
`~
`
`§
`e!i
`
`0
`N
`
`... 0
`
`.,..
`i-1B
`....
`r-1
`N
`
`1,,
`....
`N
`d
`N
`
`t
`
`..i
`N
`
`.,.;
`N
`
`~! ~ 00
`~~
`,0 " "'"'
`
`~
`!o
`g
`
`1,,
`a
`~
`
`~
`.....
`~~
`.,.,
`
`N
`
`5
`
`10
`
`15
`
`20
`
`25
`
`30
`
`35
`
`40
`
`Two-Theta
`
`

`

`Patent Application Publication Nov. 2, 2006 Sheet 5 of 8
`
`US 2006/0247189 Al
`
`Figure 5 X-ray Powder Diffraction Pattern of Az.acitidine, Form V, Labeled with the
`more Prominent 28 Angles (Cu Kcx Radiation)
`
`0 co
`:! N
`
`3000
`
`2500
`
`2000
`
`£ 1500
`~ QI = 1000
`-
`
`500
`
`0
`
`5
`
`15
`
`20
`
`25
`
`30
`
`35
`
`40
`
`Two-Theta
`
`

`

`Patent Application Publication Nov. 2, 2006 Sheet 6 of 8
`
`US 2006/0247189 Al
`
`Figure 6 X-ray Powder Diffraction Pattern of Azacitidine, Form VI, Labeled with the
`more Prominent 28 Angles (Cu Ka Radiation)
`
`1500
`
`1000
`
`,e,
`·;
`f
`
`o-4
`
`500
`
`0
`
`~
`!'.i
`
`;.
`a
`~
`
`:! 0
`~:
`"; ~
`
`N
`
`b
`N "'
`~
`
`~
`~
`
`~~
`~~
`N
`
`s
`l::i
`~
`'°
`
`N
`
`N
`
`00
`
`~
`~
`
`:lo
`;;i;
`~
`
`~
`00
`N
`
`00
`
`~ ,..:
`..,
`
`8
`~
`
`5
`
`10
`
`15
`
`20
`
`25
`
`30
`
`35
`
`40
`
`

`

`Patent Application Publication Nov. 2, 2006 Sheet 7 of 8
`
`US 2006/0247189 Al
`
`Figure 7 X-ray Powder Diffraction Pattern of Azacitidine, Mixed Phase Forms I and
`VII, Labeled with the more Prominent 28 Angles (Cu Ka. Radiation)
`
`• indcates lines distinctive to Form VII
`
`3000
`
`2500
`
`2000
`
`...
`.e-
`~ 1500
`-
`J:!
`=
`
`1000
`
`500
`
`0
`
`* :lo
`g, c!i
`.... - 00
`8o ~ ~
`ON
`j
`
`"':
`:;;
`
`1ll
`:e
`
`st
`
`~
`'."!
`
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`~
`~ ~ :a
`.... ~
`"1
`....
`gg
`"'
`
`~
`t::i
`
`~
`N
`gi
`&l
`"1 8
`;:l
`....
`"'
`
`00
`N
`
`*
`'°
`~
`
`N
`
`0
`
`.,..
`
`~~
`;:l"'
`
`5
`
`IO
`
`15
`
`20
`
`25
`
`30
`
`35
`
`40
`
`Two-Theta
`
`

`

`Patent Application Publication Nov. 2, 2006 Sheet 8 of 8
`
`US 2006/0247189 Al
`
`Figure 8 X-ray Powder Diffraction Pattern of Amcitidine, Form VIII, Labeled with the
`more Prominent 28 Angles (Cu Ka. Radiation)
`
`3500
`
`3000
`
`2500
`
`2000
`
`1500
`
`1000
`
`500
`
`0
`
`..,
`;t::
`~
`~
`
`-
`
`b-
`~
`.,;
`
`~ .,..
`.,..
`~
`
`~
`"'
`
`~
`:!
`~
`
`§
`~
`
`b
`
`"1
`
`"'
`~ -
`:g B :
`~
`
`~
`~ ~
`r--=
`
`-
`
`0
`
`~::;:
`~~
`
`~
`...,
`~
`
`~
`"'
`c,;
`N
`
`;!;
`N
`<'"l
`
`~
`i:i
`vi
`<'"l
`
`rs
`::l
`"'
`
`5
`
`10
`
`15
`
`20
`
`25
`
`30
`
`35
`
`40
`
`Two-Theta
`
`

`

`US 2006/0247189 Al
`
`Nov. 2, 2006
`
`1
`
`FORMS OF 5-AZACYTIDINE
`
`RELATED APPLICATIONS
`[0001] This application is a continuation of U.S. Pat. No.
`7,078,518, filed Feb. 7, 2005, entitled, "Forms of 5-Azacy(cid:173)
`tidine", which is a divisional of U.S. Pat. No. 6,887,855,
`filed Mar. 17, 2003, entitled "Forms of 5-Azacytidine". All
`of the above listed references are hereby incorporated in
`their entirety.
`
`FIELD OF THE INVENTION
`
`[0002] The invention relates to the isolation of crystalline
`polymorphic and pseudopolymorphic forms of 5-azacyti(cid:173)
`dine (also known as azacitidine and 4-amino-l-~-D-ribo(cid:173)
`furanosyl-S-triazin-2(1H)-one). 5-azacytidine may be used
`in the treatment of disease, including the treatment of
`myelodysplastic syndromes (MDS).
`
`BACKGROUND OF THE INVENTION
`
`[0003] Polymorphs exist as two or more crystalline phases
`that have different arrangements and/or different conforma(cid:173)
`tions of the molecule in a crystal lattice. When a solvent
`molecule(s) is contained within the crystal lattice the result(cid:173)
`ing crystal is called a pseudopolymorph, or solvate. If the
`solvent molecule(s) within the crystal structure is a water
`molecule, then the pseudopolymorph/solvate is called a
`hydrate. The polymorphic and pseudopolymorphic solids
`display different physical properties, including those due to
`packing, and various thermodynamic, spectroscopic, inter(cid:173)
`facial and mechanical properties (See H. Brittain, Polymor(cid:173)
`phism in Pharmaceutical Solids, Marcel Dekker, New York,
`N.Y., 1999, pp. 1-2). Polymorphic and pseudopolymorphic
`forms of the drug substance (also known as the "active
`pharmaceutical ingredient" (API)), as administered by itself
`or formulated as a drug product (also known as the final or
`finished dosage form, or as the pharmaceutical composition)
`are well known and may affect, for example, the solubility,
`stability, flowability, fractability, and compressibility of drug
`substances and the safety and efficacy of drug products, (see,
`e.g., Knapman, K Modem Drug Discoveries, March 2000:
`53).
`[0004] 5-azacytidine (also known as azacitidine and
`4-amino- l-~-D-ribofuranosyl-1,3,5-triazin-2(1H)-one;
`Nation Service Center designation NSC-102816; CAS Reg(cid:173)
`istry Number 320-67-2) has undergone NCI-sponsored
`clinical trials for the treatment of myelodysplastic syn(cid:173)
`dromes (MDS). See Komblith et al., J. Clin. Oneal. 20(10):
`2441-2452 (2002) and Silverman et al., J. Clin. Oneal.
`20(10): 2429-2440 (2002). 5-azacytidine may be defined as
`having a formula of C8H 12N4 0 5 , a molecular weight of
`244.20 and a structure of:
`
`HO
`
`OH
`
`OH
`
`The polymorphic form of 5-azacytidine drug substance and
`drug product has never been characterized. It is an object of
`the present invention to characterize the polymorphic forms
`of 5-azacytidine.
`
`SUMMARY OF THE INVENTION
`
`[0005]
`It has been unexpectedly found that 5-azacytidine
`in at
`least eight different polymorphic and
`exists
`pseudopolymorphic crystalline forms (Forms I-VIII), in
`addition to an amorphous form. Form I is a polymorph found
`in prior art retained samples of 5-azacytidine drug substance.
`Form II is a polymorph found in some prior art retained
`samples of the 5-azacytidine drug substance; in those
`samples, Form II is always found in mixed phase with Form
`I. Form III is a hydrate, and is formed when prior art retained
`and current samples of the drug product are reconstituted
`with water to form a "slurry" prior to administration to the
`patient. Form VI is found in prior art retained samples of the
`5-azacytidine drug product, either substantially free of other
`polymorphs, or in mixed phase with Form I.
`
`[0006] The invention provides novel crystalline forms
`referred to as Form IV, Form V, Form VII and Form VIII.
`Forms I-VIII each have characteristic X-ray power diffrac(cid:173)
`tion (XRPD) patterns and are easily distinguished from one
`another using XRPD.
`[0007] Also included in the present invention are methods
`for robustly and reproducibly synthesizing 5-azacytidine
`drug substance substantially as Fom1 IV, Form V, or Fom1
`VIII. Also provided are methods for robustly and reproduc(cid:173)
`ibly synthesizing a Form I/VII mixed phase. The invention
`also provides pharmaceutical compositions comprising the
`various forms of 5-azacytidine together with one or more
`pharmaceutically acceptable excipients, diluents, or carriers.
`
`BRIEF DESCRIPTION OF THE FIGURES
`
`[0008] FIG. 1 presents the X-Ray Powder Diffraction
`(XRPD) pattern of 5-azacytidine, Form I, labeled with the
`most prominent 28 angles (Cu Ka radiation).
`
`[0009] FIG. 2 presents the XRPD pattern of 5-azacytidine,
`mixed phase Form I and Form II, labeled with the most
`prominent 28 angles (Cu Ka radiation).
`[0010] FIG. 3 presents the XRPD pattern of 5-azacytidine,
`Form III, labeled with the most prominent 28 angles (Cu Ka
`radiation).
`
`[0011] FIG. 4 presents the XRPD pattern of 5-azacytidine,
`Form IV, labeled with the most prominent 28 angles (Cu Ka
`radiation).
`[0012] FIG. 5 presents the XRPD pattern of 5-azacytidine,
`Form V, labeled with the most prominent 28 angles (Cu Ka
`radiation).
`
`[0013] FIG. 6 presents the XRPD pattern of 5-azacytidine,
`Form VI, labeled with the most prominent 28 angles (Cu Ka
`radiation).
`[0014] FIG. 7 presents the XRPD pattern of 5-azacytidine,
`mixed phase Form I and Form VII, labeled with the most
`prominent 28 angles (Cu Ka radiation).
`
`[0015] FIG. 8 presents the XRPD pattern of 5-azacytidine,
`Form VIII, labeled with the most prominent 28 angles (Cu
`Ka radiation).
`
`

`

`US 2006/0247189 Al
`
`Nov. 2, 2006
`
`2
`
`DETAILED DESCRIPTION OF THE
`PREFERRED EMBODIMENTS
`
`22.3° 28. Table 2 provides the most prominent 28 angles,
`d-spacing and relative intensities for this mixed phase.
`
`5-azacytidine Crystalline Farms I-VIII
`
`TABLE 2
`
`[0016]
`It has been discovered that 5-azacytidine exists in
`at least eight different polymorphic and pseudopolymorphic
`crystalline forms, and also in an amorphous form.
`
`5-azacytidine, Mixed Phase Forms I and II - the most prominent
`28 angles cl-spacing and relative intensities (Cu Ka radiation)
`
`20 Angle ( 0
`
`)
`
`cl-spacing (A)
`
`Relative Intensity
`
`[0017] Form I
`
`[0018] A single sample of the 5-azacytidine drug sub(cid:173)
`stance was synthesized from 5-azacytosine and 1,2,3,5,(cid:173)
`Tetra-O-acetyl-~-D-ribofuranose according to the prior art
`method provided in Example 1. The last step of this method
`is a recrystallization of the crude synthesis product from a
`DMSO/methanol co-solvent system. Specifically, the crude
`synthesis product is dissolved in DMSO (preheated to about
`90° C.), and then methanol is added to the DMSO solution.
`The co-solvent mixture is equilibrated at approximately
`-20° C. to allow 5-azacytidine crystal formation. The prod(cid:173)
`uct is collected by vacuum filtration and allowed to air dry.
`
`[0019] The X-Ray Powder Diffraction (XRPD; see
`Example 5) pattern of the resulting 5-azacytidine is shown
`in FIG.1 along with some of the 28 values. Table 1 provides
`the most prominent 28 angles, d-spacing and relative inten(cid:173)
`sities for this material, which is designated as Form I.
`
`TABLE 1
`
`5-azacytidine Form I - the most prominent 20 angles, cl-spacing
`and relative intensities (Cu Ka radiation)
`
`20 Angle ( 0
`
`)
`
`cl-spacing (A)
`
`Relative Intensity
`
`12.182
`13.024
`14.399
`16.470
`18.627
`19.049
`20.182
`21.329
`23.033
`23.872
`26.863
`27.135
`29.277
`29.591
`30.369
`32.072
`
`7.260
`6.792
`6.146
`5.378
`4.760
`4.655
`4.396
`4.162
`3.858
`3.724
`3.316
`3.284
`3.048
`3.016
`2.941
`2.788
`
`39.1
`44.1
`31.5
`27.1
`16.0
`35.9
`37.0
`12.4
`100.0
`28.0
`10.8
`51.5
`25.6
`11.5
`10.8
`13.4
`
`[0020] Thermal analysis of Form I indicates that this form
`of 5-azacytidine is anhydrous. See Example 6.
`
`[0021] Form II
`
`[0022] Retained samples of the drug substance previously
`used to the formulate the drug product in the NCI-sponsored
`Cancer and Leukaemia Group B (CALGB) investigations
`(Phase 2 trial 8291 and Phase 3 trial 9221) for the treatment
`of MDS (Investigational New Drug (IND) 7574) were also
`analyzed by XRPD. The retained drug substance samples
`comprised either Form I, or a mixed phase of Form I and
`another polymorph: Form II. See Example 5.
`
`[0023] The XRPD powder pattern of mixed phase Forms
`I and II is shown in FIG. 2 along with some of the 28 values.
`Peaks distinctive to Form II are observed at 13.5, 17.6 and
`
`12.244
`13.082
`13.458*
`14.452
`16.521
`17.648*
`18.677
`19.093
`20.231
`21.353
`22.309*
`23.070
`23.909
`26.641
`26.813
`27.158
`29.309
`29.609
`30.384
`32.074
`
`7.223
`6.762
`6.574
`6.124
`5.361
`5.022
`4.747
`4.645
`4.386
`4.158
`3.982
`3.852
`3.719
`3.343
`3.322
`3.281
`3.045
`3.015
`2.939
`2.788
`
`34.8
`37.0
`29.2
`25.4
`19.0
`12.1
`12.7
`41.3
`42.1
`15.5
`35.1
`100.0
`18.9
`18.2
`12.6
`46.0
`27.3
`12.7
`10.5
`12.0
`
`[0024] These results indicate that the prior art 5-azacyti(cid:173)
`dine synthesis procedures for the drug substance produce
`either Form I substantially free of other forms, or a Form I/II
`mixed phase i.e. a solid material in which 5-azacytidine is
`present in a mixed phase of both Form I and Form II.
`[0025] Thermal analysis of mixed phase Form I/II is
`presented in Example 6.
`[0026] Form III
`[0027] An additional crystalline form of 5-azacytidine,
`designated Form III, is found in slurries of 5-azacytidine.
`See Example 8. Moreover, it has been found that all forms
`of 5-azacytidine (including the 5-azacytidine in the prior art
`drug product) convert to Form III in water. See Example 8.
`Thus, reconstitution of the drug product used in the afore(cid:173)
`mentioned NCI trials would have led to the formation of a
`saturated solution ( or "slurry") in which the remaining solid
`5-azacytidine was Form III. The XRPD powder pattern of
`Form III is shown in FIG. 3 along with some of the 28
`values. Table 3 provides the most prominent 28 angles,
`d-spacing and relative intensities for this crystalline mate(cid:173)
`rial. The XRPD powder pattern for Form III is distinctly
`different from that of all of the other forms of 5-azacytidine.
`
`TABLE 3
`
`5-azacytidine, Form III - the most prominent 20 angles, cl-spacing
`and relative intensities (Cu Ka radiation)
`
`20 Angle ( 0
`
`)
`
`cl-spacing (A)
`
`Relative Intensity
`
`6.566
`11.983
`13.089
`15.138
`17.446
`20.762
`21.049
`22.776
`24.363
`
`13.450
`7.380
`6.758
`5.848
`5.079
`4.275
`4.147
`3.901
`3.651
`
`32.9
`52.5
`71.0
`38.9
`48.2
`10.8
`34.8
`89.5
`13.7
`
`

`

`US 2006/0247189 Al
`
`Nov. 2, 2006
`
`3
`
`TABLE 3-continued
`
`5-azacyticline, Form III - the most prominent 20 angles, cl-spacing
`and relative intensities (Cu Ka radiation)
`
`20 Angle ( 0
`
`)
`
`cl-spacing (A)
`
`Relative Intensity
`
`25.743
`26.305
`28.741
`31.393
`32.806
`33.043
`33.536
`36.371
`39.157
`41.643
`
`3.458
`3.385
`3.104
`2.847
`2.728
`2.709
`2.670
`2.468
`2.299
`2.167
`
`22.8
`39.9
`100.0
`22.5
`11.8
`10.1
`15.1
`11.0
`19.3
`12.1
`
`[0028] Thermal analysis and proton (1H) NMR spectros(cid:173)
`copy indicate that Form III is a pseudopolymorphic form of
`5-azacytidine, specifically a monohydrate. See Examples
`6-7.
`[0029] Form IV
`[0030] Form IV is a novel crystalline form of 5-azacyti(cid:173)
`dine. Form IV was recovered by slow recrystallization from
`a DMSO/toluene co-solvent system (see Example 2) or by
`fast recrystallization from the DMSO/chloroform co-solvent
`system (see Example 3). The XRPD powder pattern of Form
`IV is shown in FIG. 4 along with some of the 28 values.
`Table 4 provides the most prominent 28 angles, d-spacing
`and relative intensities for this crystalline material. The
`XRPD powder pattern for Form IV is distinctly different
`from that of any other form.
`
`TABLE 4
`
`5-azacyticline Form IV - the most prominent 20 angles, cl-spacing
`and relative intensities (Cu Ka radiation)
`
`20 Angle ( 0
`
`)
`
`cl-spacing (A)
`
`Relative Intensity
`
`5.704
`11.571
`12.563
`14.070
`15.943
`16.993
`18.066
`20.377
`20.729
`21.484
`21.803
`22.452
`22.709
`23.646
`24.068
`25.346
`25.346
`26.900
`27.991
`28.527
`28.723
`30.124
`30.673
`31.059
`35.059
`38.195
`38.403
`
`15.408
`7.642
`7.040
`6.289
`5.555
`5.213
`4.906
`4.355
`4.281
`4.132
`4.073
`3.957
`3.913
`3.760
`3.695
`3.526
`3.511
`3.312
`3.185
`3.126
`3.106
`2.964
`2.912
`2.877
`2.557
`2.354
`2.342
`
`24.9
`97.8
`22.2
`100.0
`67.4
`51.0
`20.1
`44.7
`49.0
`36.30
`11.2
`66.7
`64.0
`17.3
`19.4
`12.0
`12.5
`11.0
`11.4
`25.7
`34.1
`14.7
`53.6
`15.7
`18.1
`15.0
`12.6
`
`[0031] Thermal analysis of Form IV is presented
`Example 6.
`
`in
`
`[0032] Form V
`
`[0033] Form V is a novel crystalline form of 5-azacyti(cid:173)
`dine. Form V was recovered by fast recrystallization of
`5-azacytidine from a DMSO/toluene co-solvent system (see
`Example 3). The XRPD powder pattern of Form Vis shown
`in FIG. 5 along with some of the 28 values. Table 5 provides
`the most prominent 28 angles, d-spacing and relative inten(cid:173)
`sities for this crystalline material. The XRPD powder pattern
`for Form Vis distinctly different from that of any other form.
`
`TABLE 5
`
`5-azacyticline Form V - the most prominent 20 angles, cl-spacing
`and relative intensities (Cu Ka radiation)
`
`20 Angle ( 0
`
`)
`
`cl-spacing (A)
`
`Relative Intensity
`
`11.018
`12.351
`13.176
`13.747
`14.548
`15.542
`16.556
`17.978
`18.549
`19.202
`19.819
`20.329
`21.518
`21.970
`22.521
`23.179
`24.018
`24.569
`27.224
`28.469
`29.041
`29.429
`30.924
`31.133
`37.938
`
`8.024
`7.160
`6.714
`6.436
`6.084
`5.697
`5.350
`4.930
`4.780
`4.618
`4.476
`4.365
`4.126
`4.042
`3.948
`3.834
`3.702
`3.620
`3.273
`3.133
`3.072
`3.033
`2.889
`2.870
`2.370
`
`40.0
`29.6
`28.3
`42.9
`18.3
`14.2
`47.8
`18.1
`83.9
`25.0
`12.1
`28.6
`100.0
`65.6
`11.5
`66.5
`13.0
`40.7
`50.2
`24.2
`24.8
`15.0
`15.6
`22.6
`10.7
`
`[0034] Thermal analysis indicates that Form Vis a solvate.
`See Example 6.
`
`[0035] Form VI
`
`[0036] The drug product used in the aforementioned NCI
`investigation was typically prepared by lypohilizing a solu(cid:173)
`tion of 5-azacytidine and mamiitol (1:1 w/w). The resultant
`drug product comprised 100 mg of 5-azacytidine and 100
`mg mamiitol as a lyophilized cake in a vial and was
`administered by subcutaneous injection as an aqueous sus(cid:173)
`pension ("slurry"). XRPD analysis of retained samples of
`the drug product used in the NCI investigation revealed the
`existence of another polymorph, Form VI. The retained drug
`product samples comprised either Form VI alone, or a Form
`I/VI mixed phase. Table 6 provides the most prominent 28
`angles, d-spacing and relative intensities for Form VI.
`
`TABLE 6
`
`5-azacyticline Form VI - the most prominent 20 angles, cl-spacing
`and relative intensities (Cu Ka radiation)
`
`20 Angle ( 0
`
`)
`
`cl-spacing (A)
`
`Relative Intensity
`
`12.533
`12.963
`13.801
`
`7.057
`6.824
`6.411
`
`10.1
`10.2
`100.0
`
`

`

`US 2006/0247189 Al
`
`Nov. 2, 2006
`
`4
`
`TABLE 6-continued
`
`5-azacyticline Form VI - the most prominent 20 angles, cl-spacing
`and relative intensities (Cu Ka radiation)
`
`20 Angle ( 0
`
`)
`
`cl-spacing (A)
`
`Relative Intensity
`
`18.929
`20.920
`21.108
`21.527
`22.623
`22.970
`24.054
`26.668
`27.210
`28.519
`29.548
`30.458
`33.810
`35.079
`37.528
`
`4.6843
`4.243
`4.205
`4.125
`3.922
`3.869
`3.697
`3.340
`3.275
`3.127
`3.021
`2.932
`2.649
`2.556
`2.411
`
`10.0
`34.2
`49.4
`47.0
`10.7
`13.8
`77.8
`23.0
`33.7
`12.9
`27.2
`50.3
`11.6
`12.6
`24.7
`
`[0037] Thermal analysis and proton (1H) NMR spectros(cid:173)
`copy of Form VI is presented in Examples 6-7.
`[0038] Form VII
`[0039] Form VII is a novel crystalline form of 5-azacyti(cid:173)
`dine. Form VII was produced by fast recrystallization from
`a DMSO/methanol co-solvent system (see Example 3).
`Form VII was always isolated by this recrystallization
`method as a mixed phase with Form I. The XRPD powder
`pattern of mixed phase Forms I and VII is shown in FIG. 7
`along with some of the 28 values and the Form VII distinc(cid:173)
`tive peaks indicated with asterisks. Table 7 provides the
`most prominent 28 angles, d-spacing and relative intensities
`for this mixed phase. Form VII exhibits distinctive peaks at
`5.8, 11.5, 12.8, 22.4 and 26.6° 28 in addition to peaks
`displayed in the Form I XRPD powder pattern. The XRPD
`pattern for mixed phase Forms I and VII is distinctly
`different from that of any other form.
`
`TABLE 7
`
`5-azacytidine, mixed Forms I and VII - the most prominent
`28 angles cl-spacing and relative intensities (Cu Ka radiation)
`
`20 Angle ( 0
`
`)
`
`cl-spacing (A)
`
`Relative Intensity
`
`5.779
`11.537
`12.208
`12.759
`13.048
`14.418
`16.489
`18.649
`19.101
`20.200
`20.769
`21.355
`22.365
`23.049
`23.884
`26.628
`27.145
`29.296
`29.582
`32.078
`
`15.281
`7.664
`7.244
`6.932
`6.780
`6.138
`5.372
`4.754
`4.643
`4.392
`4.273
`4.157
`3.972
`3.856
`3.723
`3.345
`3.282
`3.046
`3.017
`2.788
`
`14.7
`8.3
`28.0
`21.7
`34.4
`22.5
`21.6
`13.5
`34.7
`34.4
`10.5
`11.7
`29.9
`100.0
`23.1
`13.3
`52.9
`26.2
`11.3
`12.9
`
`[0041] Form VIII
`
`[0042] Form VIII is a novel crystalline form of 5-azacy(cid:173)
`tidine. Form VIII was recovered by recrystallizing 5-azacy(cid:173)
`tidine Form I from a N-methyl-2-pyrrolidone (NMP) single
`solvent system (see Example 4). The XRPD powder pattern
`of Form VIII is shown in FIG. 8 along with some of the 28
`values. Table 8 provides the most prominent 28 angles,
`d-spacing and relative intensities for this material. The
`XRPD pattern for Form VIII is distinctly different from that
`of any other form.
`
`TABLE 8
`
`5-azacyticline, Form VIII - the most prominent 20 angles,
`cl-spacing and relative intensities (Cu Ka radiation)
`
`20Angle ( 0
`
`)
`
`cl-spacing (A)
`
`Relative Intensity
`
`6.599
`10.660
`12.600
`13.358
`15.849
`17.275
`20.243
`20.851
`21.770
`22.649
`25.554
`25.740
`29.293
`32.148
`35.074
`38.306
`
`13.384
`8.292
`7.020
`6.623
`5.587
`5.129
`4.383
`4.257
`4.079
`3.923
`3.483
`3.458
`3.046
`2.782
`2.556
`2.348
`
`2.9
`2.2
`23.4
`2.6
`2.0
`4.2
`5.8
`7.8
`74.4
`32.1
`100.0
`7.8
`3.8
`8.8
`7.4
`2.5
`
`[0043] Amorphous 5-azacytidine
`
`[0044] Amorphous 5-azacytidine may be recovered from
`equilibrium saturated solutions of 5-azacytidine in propy(cid:173)
`lene glycol, polyethylene glycol and DMSO. See Example
`8.
`
`Pharmaceutical Formulations
`
`[0045] For the most effective administration of drug sub(cid:173)
`stance of the present invention, it is preferred to prepare a
`pharmaceutical formulation (also known as the "drug prod(cid:173)
`uct") preferably in unit dose form, comprising one or more
`of the 5-azacytidine forms of the present invention and one
`or more pharmaceutically acceptable carrier, diluent, or
`excipient.
`
`[0046] Such pharmaceutical formulation may, without
`being limited by the teachings set forth herein, include a
`solid form of the present invention which is blended with at
`least one pharmaceutically acceptable excipient, diluted by
`an excipient or enclosed within such a carrier that can be in
`the form of a capsule, sachet, tablet, buccal, lozenge, paper,
`or other container. When the excipient serves as a diluent, it
`may be a solid, semi-solid, or liquid material which acts as
`a vehicle, carrier, or medium for the 5-azacytidine polymor(cid:173)
`ph( s ). Thus, the formulations can be in the form of tablets,
`pills, powders, elixirs, suspensions, emulsions, solutions,
`syrups, capsules (such as, for example, soft and hard gelatin
`capsules), suppositories, sterile injectable solutions, and
`sterile packaged powders.
`
`[0040] Thermal analysis of Form VII is presented in
`Example 6
`
`[0047] Examples of suitable excipients include, but are not
`limited to, starches, gum arabic, calcium silicate, microc-
`
`

`

`US 2006/0247189 Al
`
`Nov. 2, 2006
`
`5
`
`rystalline cellulose, polyvinylpyrrolidone, cellulose, water,
`syrup, and methyl cellulose. The formulations can addition(cid:173)
`ally include lubricating agents such as, for example, talc,
`magnesium stearate and mineral oil; wetting agents; emul(cid:173)
`sifying and suspending agents; preserving agents such as
`methyl- and propyl-hydroxybenzoates; sweetening agents;
`or flavoring agents. Polyols, buffers, and inert fillers may
`also be used. Examples of polyols include, but are not
`limited to: mannitol, sorbitol, xylitol, sucrose, maltose,
`glucose, lactose, dextrose, and the like. Suitable buffers
`encompass, but are not limited to, phosphate, citrate, tar(cid:173)
`trate, succinate, and the like. Other inert fillers which may be
`used encompass those which are known in the art and are
`useful in the manufacture of various dosage forms. If
`desired, the solid pharmaceutical compositions may include
`other components such as bulling agents and/or granulating
`agents, and the like. The compositions of the invention can
`be formulated so as to provide quick, sustained, controlled,
`or delayed release of the drug substance after administration
`to the patient by employing procedures well known in the
`art.
`
`[0048]
`In certain embodiments of the invention, the 5-aza(cid:173)
`cytidine forms( s) may be made into the form of dosage units
`for oral administration. The 5-azacytidine forms(s) may be
`mixed with a solid, pulverant carrier such as, for example,
`lactose, saccharose, sorbitol, mannitol, starch, amylopectin,
`cellulose derivatives or gelatin, as well as with an antifric(cid:173)
`tion agent such as for example, magnesium stearate, calcium
`stearate, and polyethylene glycol waxes. The mixture is then
`pressed into tablets or filled into capsules. If coated tablets,
`capsules, or pulvules are desired, such tablets, capsules, or
`pulvules may be coated with a concentrated solution of
`sugar, which may contain gum arabic, gelatin, talc, titanium
`dioxide, or with a lacquer dissolved in the volatile organic
`solvent or mixture of solvents. To this coating, various dyes
`may be added in order to distinguish among tablets with
`different active compounds or with different amounts of the
`active compound present.
`
`[0049] Soft gelatin capsules may be prepared in which
`capsules contain a mixture of the 5-azacytidine form(s) and
`vegetable oil or non-aqueous, water miscible materials such
`as, for example, polyethylene glycol and the like. Hard
`gelatin capsules may contain granules or powder of the
`5-azacytidine polymorph in combination with a solid, pul(cid:173)
`verulent carrier, such as, for example, lactose, saccharose,
`sorbitol, mannitol, potato starch, corn starch, amylopectin,
`cellulose derivatives, or gelatin.
`
`[0050] Tablets for oral use are typically prepared in the
`following manner, although other techniques may be
`employed. The solid substances are gently ground or sieved
`to a desired particle size, and a binding agent is homog(cid:173)
`enized and suspended in a suitable solvent. The 5-azacyti(cid:173)
`dine form(s) and auxiliary agents are mixed with the binding
`agent solution. The resulting mixture is moistened to form a
`uniform suspension. The moistening typically causes the
`particles to aggregate slightly, and the resulting mass is
`gently pressed through a stainless steel sieve having a
`desired size. The layers of the mixture are then dried in
`controlled drying units for a pre-determined length of time
`to achieve a desired particle size and consistency. The
`granules of the dried mixture are gently sieved to remove
`any powder. To this mixture, disintegrating, anti-friction,
`and anti-adhesive agents are added. Finally, the mixture is
`
`pressed into tablets using a machine with the appropriate
`punches and dies to obtain the desired tablet size.
`
`[0051]
`In the event that the above formulations are to be
`used for parenteral administration, such a formulation typi(cid:173)
`cally comprises sterile, aqueous and non-aqueous injection
`solutions comprising one or more 5-azacytidine forms for
`which preparations are preferably isotonic with the blood of
`the intended recipient. These preparations may contain anti(cid:173)
`oxidants, buffers, bacteriostats, and solute; which render the
`formulation isotonic with the blood of the intended recipi(cid:173)
`ent. Aqueous and non-aqueous suspensions may include
`suspending agents and thickening agents. The formulations
`may be present in unit-dose or multi-dose containers, for
`example, sealed ampules and vials. Extemporaneous injec(cid:173)
`tion solutions and suspensions may be prepared from sterile
`powders, granules, and tablets of the kind previously
`described.
`
`[0052] Liquid preparations for oral administration are pre(cid:173)
`pared in the form of solutions, syrups, or suspensions with
`the latter two forms containing, for example, 5-azacytidine
`polymorph(s), sugar, and a mixture of ethanol, water, glyc(cid:173)
`erol, and propylene glycol. If desired, such liquid prepara(cid:173)
`tions contain coloring agents, flavoring agents, and saccha(cid:173)
`rin. Thickening agents such as carboxymethylcellulose may
`also be used.
`
`[0053] As such, the pharmaceutical formulations of the
`present invention are preferably prepared in a unit dosage
`form, each dosage unit containing from about 5 mg to about
`200 mg, more usually about 100 mg of the 5-azacytidine
`form(s ). In liquid form, dosage unit contains from about 5 to
`about 200 mg, more usually about 100 mg of the 5-azacy(cid:173)
`tidine form(s). The term "unit dosage form" refers to physi(cid:173)
`cally discrete units suitable as unitary dosages for human
`subjects/patients or other mammals, each unit containing a
`predetermined quantity of the 5-azacytidine polymorph cal(cid:173)
`culated to produce the desired therapeutic effect, in associa(cid:173)
`tion with preferably, at least one pharmaceutically accept(cid:173)
`able carrier, diluent, or excipient.
`
`[0054] The following examples are provided for illustra(cid:173)
`tive purposes only, and are not to be construed as limiting the
`scope of the claims in any way.
`
`EXAMPLES
`
`Example 1
`
`Prior Art Procedure for Synthesis of 5-azacytidine
`Drug Substance
`
`[0055] Using commercially available 5-azacytosine (1)
`and 1,2,3,5-Tetra-O-~-acetyl-ribofuranose (2) (RTA), 5-aza(cid:173)
`cytidine (3) may be synthesized according to the pathway
`below.
`
`(1)
`
`

`

`US 2006/0247189 Al
`
`Nov. 2, 2006
`
`6
`
`-continued
`
`+
`
`A cO~ OAc
`0
`H
`H
`
`H
`
`OAc
`(2)
`
`H
`
`OAc
`
`solvent, preheated to approximately 90° C., in separate
`100-ml beakers. The solids were allowed to dissolve to a
`clear solution. Approximately 45 mL of the selected co(cid:173)
`solvent (methanol, toluene, or chloroform), preheated to
`approximately 50° C., was added to the solution and the
`resultant solution was mixed. The solution was covered and
`placed in a freezer to equilibrate at approximately -20° C.
`to allow crystal formation. Solutions were removed from the
`freezer after crystal formation.
`
`[0059] The product from the methanol and toluene solu(cid:173)
`tions was collected by vacuum filtration using a Buchner
`funnel. The resulting white crystalline product was allowed
`to air dry.
`
`[0060] The chloroform product was too fine to be col(cid:173)
`lected by vacuum filtration. Most of the solvent was care(cid:173)
`fully decanted from the chloroform solution and the solvent
`from the resultant slurry was allowed to evaporate at ambi(cid:173)
`ent temperature to dryness. The chloroform solution evapo(cid:173)
`rated to a white product. Note that fast recrystallization
`using the DMSO/methanol co-solvent sys