(12) INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT)
`
`(19) World Intellectual Property
`Organization
`International Bureau
`
`Mill N il I
`
`(43) International Publication Date
`30 September 2004 (30.09.2004)
`
`PCX
`
`(10) International Publication N u m b er
`
`WO 2004/082619 A2
`
`(51) International Patent Classification7:
`
`A61K
`
`(21) International Application Number:
`
`(74) Agents: HIRD, Steven et a l; Swanson & Bratschun LLC,
`1745 Shea Center Drive, Suite 330, Highlands Ranch, CO
`80129 (US).
`
`(22) International Filing Date: 16 March 2004 (16.03.2004)
`
`PCT/US2004/007895
`
`(25) Filing Language:
`
`(26) Publication Language:
`(30) Priority Data:
`10/390,578
`
`English
`
`English
`
`17 March 2003 (17.03.2003)
`
`US
`
`(71) Applicants
`except US):
`(for all de.signate.d States
`PHARMION CORPORATION
`[US/US]; 2525 28th
`Street, Boulder, CO 80301 (US). ASH STEVENS INC
`[US/US]; 5861 John C Lodge Freeway, Detroit, MI 48202
`(US).
`
`(72) Inventors; and
`IONESCU, Du-
`(75) Inventors/Applicants (for US only):
`mitru [RO/US]; 2828 Grant Drive, Ann Arbor, MI 48108
`(US). BLUMBERGS, Peter [US/US]; 4105 Springer,
`Royal Oak, MI 48073 (US). SILVEY, Gary, L. [US/US];
`10139 Switzer Circle, Overland Park, KS 66212 (US).
`
`(54) Title: FORMS OF 5-AZACYTIDINE
`
`(81) Designated States (unless otherwise
`indicated, for every
`kind of national protection available): AE, AG, AL, AM,
`AT, AU, AZ, BA, BB, BG, BR, BW, BY, BZ, CA, CH, CN,
`CO, CR, CU, CZ, DE, DK, DM, DZ, EC, EE, EG, ES, FI,
`GB, GD, GE, GH, GM, HR, HU, ID, IL, IN, IS, IP, KB,
`KG, KP, KR, KZ, LC, LK, LR, LS, LT, LU, LV, MA, MD,
`MG, MK, MN, MW, MX, MZ, NA, NI, NO, NZ, OM, PG,
`PH, PL, PT, RO, RU, SC, SD, SE, SG, SK, SL, SY, TJ, TM,
`TN, TR, I T, I Z, UA, UG, US, UZ, VC, VN, YU, ZA, ZM,
`ZW.
`
`(84) Designated States (unless otherwise
`indicated, for every
`kind of regional protection available): ARIPO (BW, GH,
`GM, KE, LS, MW, MZ, SD, SL, SZ, TZ, UG, ZM, ZW),
`Eurasian (AM, AZ, BY, KG, KZ, MD, RU, TJ, TM), Euro(cid:173)
`pean (AT, BE, BG, CH, CY, CZ, DE, DK, EE, ES, El, FR,
`GB, GR, HU, IE, IT, LU, MC, NL, PL, PT, RO, SE, SI, SK,
`TR), OAPI (BE, BJ, CF, CG, CI, CM, GA, GN, GQ, GW,
`ML, MR, NE, SN, TD, TG).
`
`[Continued on next page]
`
`Fignre 1.
`
`Z-sray Psw/dter ESffiractioa iPattern af Asadtidiae, F a na 1, Labeled with the
`more PnnBdHsni 28 Amfles (Co Res HaifeftBm)
`
`1 i "i
`
`|
`
`i
`
`i > > \
`
`i
`
`i
`
`i f
`
`|
`
`i
`
`t' t
`
`i
`
`\
`
`i
`
`i
`
`i i
`
`|
`
`i
`
`i
`
`i
`
`i [ i
`
`i
`
`i
`
`i
`
`|
`
`t
`
`t
`
`i
`
`i )
`
`5500
`
`5000
`
`4500
`
`4000
`
`3500
`
`3000
`
`1 2500
`
`5 2000
`
`1500
`
`1000
`
`500
`
`0
`
`<
`
`''
`
`•
`
`'
`
`'
`
`*
`
`•
`
`'
`
`*
`
`•
`
`•
`
`•
`
`*
`
`'
`
`'
`
`•
`
`*
`
`*
`
`'
`
`'
`
`'
`
`•
`
`*
`
`'
`
`•
`
`'
`
`'
`
`»
`
`*
`
`10
`
`15
`
`20
`
`25
`
`30
`
`35
`
`•
`
`•
`
`'
`
`•
`
`40
`
`Two-Theta
`
`O
`
`(57) Abstract: The invention provides novel polymorphic and pseudopolymorphic crystalline forms of 5-azacytidine, along with
`methods for preparing said forms, wherein 5-azacythidine is represented by the formula (I). The invention also includes pharmaceu(cid:173)
`tical compositions comprising said forms.
`
`Apotex v. Cellgene - IPR2023-00512
`Petitioner Apotex Exhibit 1004-0001
`
`

`

`WO 2 0 0 4 / 0 8 2 6 19 A2
`
`I lllllll II11 111 III III III II III lllll III I II Mill 111 III llll 111 llll
`
`Published:
`— without international search report and to be republished
`upon receipt of that report
`
`For two-letter codes and other abbreviations, refer to the "Guid-
`ance Notes on Codes and Abbreviations" appearing at the begin-
`ning of each regular issue of the PCT Gazette.
`
`Apotex v. Cellgene - IPR2023-00512
`Petitioner Apotex Exhibit 1004-0002
`
`

`

`WO 2004/082619
`
`PCT7US2004/007895
`
`5
`
`FORMS OF 5-AZACYTIDINE
`
`FIELD OF THE INVENTION
`
`The invention relates to the isolation of crystalline polymorphic and pseudopolymorphic
`
`forms of 5-azacytidine (also known as azacitidine and 4-amino-l-P-D-ribofuranosyl-S-triazin-
`
`10
`
`2(lH)-one). 5-azacytidine may be used in the treatment of disease, including the treatment of
`
`myelodysplastic syndromes (MDS).
`
`BACKGROUND OF THE INVENTION
`
`Polymorphs exist as two or more crystalline phases that have different arrangements
`
`15
`
`and/or different conformations of the molecule in a crystal lattice. When a solvent molecule(s)
`
`is contained within the crystal lattice the resulting 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
`
`20
`
`thermodynamic, spectroscopic, interfacial and mechanical properties (See H. Brittain,
`
`Polymorphism in Pharmaceutical Solids, Marcel Dekker, New York, NY, 1999, pp. 1-2).
`
`Polymorphic and pseudopolymorphic forms of the drag 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
`
`25 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.,
`
`Knapmari, K Modem Drug Discoveries, March 2000: 53).
`
`5-azacytidine (also known as azacitidine and 4-amino-l-P-D-ribofuranosyl-l,3,5-triazm-
`
`2(1/0-0116; Nation Service Center designation NSC-102816; CAS Registry Number 320-67-2)
`
`30
`
`has undergone NCI-sponsored clinical trials for the treatment of myelodysplastic syndromes
`
`(MDS). See Komblith et a!., J. Clin. Oncol. 20(10): 2441-2452 (2002) and Silverman et al, J.
`
`Clin. Oncol. 20(10): 2429-2440 (2002). 5-azacytidine may be defined as having a formula of
`
`C8H12N4O5, a molecular weight of 244.20 and a structure of:
`
`Apotex v. Cellgene - IPR2023-00512
`Petitioner Apotex Exhibit 1004-0003
`
`

`

`WO 2004/082619
`
`PCT/US2004/007895
`
`NH2
`
`M1
`Ht
`
`ft
`
`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.
`
`10
`
`SUMMARY OF THE INVENTION
`
`It has been unexpectedly found that 5-azacytidine exists in at least eight different
`
`polymorphic and 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-
`
`15
`
`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.
`
`20
`
`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 diffraction (XRPD)
`
`patterns and are easily distinguished from one another using XRPD.
`
`Also included in the present invention are methods for robustly and reproducibly
`
`synthesizing 5-azacytidine drug substance substantially as Form IV, Form V, or Form VIII.
`
`25 Also provided are methods for robustly and reproducibly 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.
`
`Apotex v. Cellgene - IPR2023-00512
`Petitioner Apotex Exhibit 1004-0004
`
`

`

`WO 2004/082619
`
`PCT/US2004/007895
`
`5
`
`BRIEF DESCRIPTION OF THE FIGURES
`
`Figure I presents the X-Ray Powder Diffraction (XRPD) pattern of 5-azacytidine, Form I,
`
`labeled with the most prominent 29 angles (Cu Ka radiation).
`
`Figure 2 presents the XRPD pattern of 5-azacytidine, mixed phase Form I and Form II,
`
`10
`
`labeled with the most prominent 29 angles (Cu Ka radiation).
`
`Figure 3 presents the XRPD pattern of 5-azacytidine, Form III, labeled with the most
`
`prominent 29 angles (Cu Ka radiation).
`
`Figure 4 presents the XRPD pattern of 5-azacytidine, Form IV, labeled with the most
`
`prominent 29 angles (Cu Ka radiation).
`
`15
`
`Figure 5 presents the XRPD pattern of 5-azacytidine, Form V, labeled with the most
`
`prominent 29 angles (Cu Ka radiation).
`
`Figure 6 presents the XRPD pattern of 5-azacytidine, Form VI, labeled with the most
`
`prominent 29 angles (Cu Ka radiation).
`
`Figure 7 presents the XRPD pattern of 5-azacytidine, mixed phase Form I and Form VII,
`
`20
`
`labeled with the most prominent 29 angles (Cu Ka radiation).
`
`Figure 8 presents the XRPD pattern of 5-azacytidine, Form VIII, labeled with the most
`
`prominent 29 angles (Cu Ka radiation).
`
`- 3-
`
`Apotex v. Cellgene - IPR2023-00512
`Petitioner Apotex Exhibit 1004-0005
`
`

`

`WO 2004/082619
`
`PCT7US2004/007895
`
`5 DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS
`
`5-azacytidine Crystalline Forms I-VIII
`
`It has been discovered that 5-azacytidine exists in at least eight different polymorphic and
`
`pseudopolymorphic crystalline forms, and also in an amorphous form.
`
`10
`
`Form I
`
`A single sample of the 5-azacytidine drug substance was synthesized from 5-azacytosine
`
`and 1,2,3,5,-Tetra-O-acetyl-P-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
`
`15
`
`from a DMSO/methanol co-solvent system. Specifically, the crude synthesis product is
`dissolved in DMSO (preheated to about 90oC), and then methanol is added to the DMSO
`solution. The co-solvent mixture is equilibrated at approximately -20oC to allow 5-azacytidine
`crystal formation. The product is collected by vacuum filtration and allowed to air dry.
`
`The X-Ray Powder Diffraction (XRPD; see Example 5) pattern of the resulting 5-
`
`azacytidine is shown in Figure 1 along with some of the 20 values. Table 1 provides the most
`
`20
`
`prominent 20 angles, d-spacing and relative intensities for this material, which is designated as
`
`Form I.
`
`29 Angle (0)
`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
`
`d-spacing (A)
`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
`
`Relative Intensity
`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
`
`Apotex v. Cellgene - IPR2023-00512
`Petitioner Apotex Exhibit 1004-0006
`
`

`

`WO 2004/082619
`
`PCT/US2004/007895
`
`5
`
`Table 1: 5-azacytidine Form I - the most prominent 28 angles, d-spacing and relative
`
`intensities (Cu Ka radiation)
`
`Thermal analysis of Form I indicates that this form of 5-azacytidine is anhydrous. See
`
`Example 6.
`
`10
`
`Form II
`
`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 Drag (IND)
`
`7574) were also analyzed by XRPD. The retained drug substance samples comprised either
`
`15
`
`Form I, or a mixed phase of Form I and another polymorph: Form II. See Example 5.
`
`The XRPD powder pattern of mixed phase Forms I and II is shown in Figure 2 along with
`some of the 20 values. Peaks distinctive to Form II are observed at 13.5, 17.6 and 22.3 0 20.
`
`Table 2 provides the most prominent 26 angles, d-spacing and relative intensities for this
`
`mixed phase.
`
`26 Angle (0)
`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
`
`d-spacing (A)
`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
`
`Relative Intensity
`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
`
`Apotex v. Cellgene - IPR2023-00512
`Petitioner Apotex Exhibit 1004-0007
`
`

`

`WO 2004/082619
`
`PCT7US2004/007895
`
`5
`
`Table 2: 5-azacytidine, Mixed Phase Forms I and II - the most prominent 29 angles, d-
`
`spacing and relative intensities (Cu Ka radiation)
`
`These results indicate that the prior art 5-azacytidine synthesis procedures for the drug
`
`substance produce either Form I substantially free of other forms, or a Form I/II mixed phase
`
`10
`
`/. e. a solid material in which 5-azacytidine is present in a mixed phase of both Form I and
`
`Form II.
`
`Thermal analysis of mixed phase Form I/II is presented in Example 6.
`
`Form III
`
`15
`
`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 aforementioned 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 Figure 3
`
`20
`
`along with some of the 29 values. Table 3 provides the most prominent 29 angles, d-spacing
`
`and relative intensities for this crystalline material. The XRPD powder pattern for Form III is
`
`distinctly different from that of all of the other forms of 5-azacytidine.
`
`29 Angle (0)
`6.566
`11.983
`13.089
`15.138
`17.446
`20.762
`21.049
`22.776
`24.363
`25.743
`26.305
`28.741
`31.393
`32.806
`33.043
`
`d-spacing (A)
`13.450
`7.380
`6.758
`5.848
`5.079
`4.275
`4.147
`3.901
`3.651
`3.458
`3.385
`3.104
`2.847
`2.728
`•2.709
`6-
`
`Relative Intensity
`32.9
`52.5
`71.0
`38.9
`48.2
`10.8
`34.8
`89.5
`13.7
`22.8
`39.9
`100.0
`22.5
`11.8
`10.1
`
`Apotex v. Cellgene - IPR2023-00512
`Petitioner Apotex Exhibit 1004-0008
`
`

`

`WO 2004/082619
`
`PCT7US2004/007895
`
`33.536
`36.371
`39.157
`41.643
`
`2.670
`2.468
`2.299
`2.167
`
`15.1
`11.0
`19.3
`12.1
`
`Table 3: 5-azacytidine5 Form III - the most prominent 29 angles, d-spacing and relative
`
`intensities (Cu Ka radiation)
`
`Thermal analysis and proton (1H) NMR spectroscopy indicate that Form III is a
`pseudopolymorphic form of 5-azacytidine, specifically a monohydrate. See Examples 6-7.
`
`10
`
`Form IV
`
`Form IV is a novel crystalline form of 5-azacytidine. 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
`
`15
`
`powder pattern of Form IV is shown in Figure 4 along with some of the 29 values. Table 4
`
`provides the most prominent 20 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.
`
`20 Angle (0)
`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
`
`d-spacing (A)
`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
`
`Relative Intensity
`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
`
`Apotex v. Cellgene - IPR2023-00512
`Petitioner Apotex Exhibit 1004-0009
`
`

`

`WO 2004/082619
`
`PCT/US2004/007895
`
`25.346
`25.346
`26.900
`27.991
`28.527
`28.723
`30.124
`30.673
`31.059
`35.059
`38.195
`38.403
`Table 4: 5-azacytidme Form IV - the most prominent 29 angles, d-spacing and relative
`
`3.526
`3.511
`3.312
`3.185
`3.126
`3.106
`2.964
`2.912
`2.877
`2.557
`2.354
`2.342
`
`12.0
`12.5
`11.0
`11.4
`25.7
`34.1
`14.7
`53.6
`15.7
`18.1
`15.0
`12.6
`
`intensities (Cu Kcc radiation)
`
`Thermal analysis of Form IV is presented in Example 6.
`
`Form V
`
`10
`
`Form V is a novel crystalline form of 5-azacytidine. 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 V is shown in Figure 5 along with some of the 20 values.
`
`Table 5 provides the most prominent 20 angles, d-spacing and relative intensities for this
`
`crystalline material. The XRPD powder pattern for Form V is distinctly different from that of
`
`15
`
`any other form.
`
`26 Angle Q
`11.018
`12.351
`13.176
`13.747
`14.548
`15.542
`16.556
`17.978
`18.549
`19.202
`19.819
`20.329
`
`d-spacing (A)
`8.024
`7.160
`6.714
`6.436
`6.084
`5.697
`5.350
`4.930
`4.780
`4.618
`4.476
`4.365
`
`- 8-
`
`Relative Intensity
`40.0
`29.6
`28.3
`42.9
`18.3
`14.2
`47.8
`18.1
`83.9
`25.0
`12.1
`28.6
`
`Apotex v. Cellgene - IPR2023-00512
`Petitioner Apotex Exhibit 1004-0010
`
`

`

`WO 2004/082619
`
`PCT/US2004/007895
`
`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
`
`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
`
`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
`
`Table 5: 5-azacytidine Form V - the most prominent 29 angles, d-spacing and relative
`
`intensities (Cu Ka radiation)
`
`Thermal analysis indicates that Form V is a solvate. See Example 6.
`
`10
`
`Form VI
`
`The drug product used in the aforementioned NCI investigation was typically prepared by
`
`lypohilizing a solution of 5-azacytidine and mannitol (1:1 w/w). The resultant drug product
`
`comprised 100 mg of 5-azacytidine and 100 mg mannitol as a lyophilized cake in a vial and
`
`was administered by subcutaneous injection as an aqueous suspension ("slurry"). XRPD
`
`15
`
`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 20
`
`angles, d-spacing and relative intensities for Form VI.
`
`20 Angle (0)
`12.533
`12.963
`13.801
`18.929
`20.920
`21.108
`21.527
`22.623
`22.970
`
`d-spacing (A)
`7.057
`6.824
`6.411
`4.6843
`4.243
`4.205
`4,125
`3.922
`3.869
`
`-9
`
`Relative Intensity
`10.1
`10.2
`100.0
`10.0
`34.2
`49.4
`47.0
`10.7
`13.8
`
`Apotex v. Cellgene - IPR2023-00512
`Petitioner Apotex Exhibit 1004-0011
`
`

`

`WO 2004/082619
`
`PCT7US2004/007895
`
`24.054
`26.668
`27.210
`28.519
`29.548
`30.458
`33.810
`35.079
`37.528
`
`3.697
`3.340
`3.275
`3.127
`3.021
`2.932
`2.649
`2.556
`2.411
`
`77.8
`23.0
`33.7
`12.9
`27.2
`50.3
`11.6
`12.6
`24.7
`
`Table 6: 5-azacytidine Form VI - the most prominent 29 angles, d-spacing and relative
`
`intensities (Cu Ka radiation)
`
`Thermal analysis and proton (1H) NMR spectroscopy of Form VI is presented in Examples
`6-7.
`
`10
`
`Form VII
`
`Form VII is a novel crystalline form of 5-azacytidine. 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 Figure 7 along with some of the
`
`15
`
`29 values and the Form VII distinctive peaks indicated with asterisks. Table 7 provides the
`
`most prominent 29 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 0 29 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.
`
`20
`
`29 Angle (0)
`5.779
`11.537
`12.208
`12.759
`13.048
`14.418
`16.489
`18.649
`19.101
`20.200
`
`d-spacing (A)
`15.281
`7.664
`7.244
`6.932
`6.780
`6.138
`5.372
`4.754
`4.643
`4.392
`10
`
`Relative Intensity
`14.7
`8.3
`28.0
`21.7
`34.4
`22.5
`21.6
`13.5
`34.7
`34.4
`
`Apotex v. Cellgene - IPR2023-00512
`Petitioner Apotex Exhibit 1004-0012
`
`

`

`WO 2004/082619
`
`PCT7US2004/007895
`
`10.5
`11.7
`29.9
`100.0
`23.1
`13.3
`52.9
`26.2
`11.3
`12.9
`Table 7: 5-azacytidine, mixed Forms I and VII - the most prominent 29 angles, d-spacing
`
`20.769
`21.355
`22.365
`23.049
`23.884
`26.628
`27.145
`29.296
`29.582
`32.078
`
`4.273
`4.157
`3.972
`3.856
`3.723
`3.345
`3.282
`3.046
`3.017
`2.788
`
`and relative intensities (Cu Ka radiation)
`
`Thermal analysis of Form VII is presented in Example 6.
`
`Form VIII
`
`Form VIII is a novel crystalline form of S-azacytidine. Form VIII was recovered by
`
`10
`
`recrystallizing 5-azacytidine 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 Figure 8 along
`
`with some of the 29 values. Table 8 provides the most prominent 29 angles, d-spacing and
`
`relative intensities for this material. The XRPD pattern for Form VIII is distinctly different
`
`from that of any other form.
`
`15
`
`29Angle (0)
`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
`
`d-spacing (A)
`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
`
`11 -
`
`Relative Intensity
`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
`
`Apotex v. Cellgene - IPR2023-00512
`Petitioner Apotex Exhibit 1004-0013
`
`

`

`WO 2004/082619
`
`PCT7US2004/007895
`
`5
`
`Table 8: 5-azacytidine, Form VIII - the most prominent 29 angles, d-spacing and relative
`
`intensities (Cu Ka radiation)
`
`Amorphous 5-azacytidine
`
`Amorphous 5-azacytidine may be recovered from equilibrium saturated solutions of 5-
`
`10
`
`azacytidine in propylene glycol, polyethylene glycol and DMSO. See Example 8.
`
`Pharmaceutical Formulations
`
`For the most effective administration of drug substance of the present invention, it is
`
`preferred to prepare a pharmaceutical formulation (also known as the "drug product")
`
`15
`
`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.
`
`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
`
`20
`
`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 polymorph(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),
`
`25
`
`suppositories, sterile injectable solutions, and sterile packaged powders.
`
`Examples of suitable excipients include, but are not limited to, starches, gum arable,
`
`calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, and
`
`methyl cellulose. The formulations can additionally include lubricating agents such as, for
`
`example, talc, magnesium stearate and mineral oil; wetting agents; emulsifying and
`
`30
`
`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, tartrate, succinate, and the like. Other inert fillers which may be
`- 1 2-
`
`Apotex v. Cellgene - IPR2023-00512
`Petitioner Apotex Exhibit 1004-0014
`
`

`

`WO 2004/082619
`
`PCT7US2004/007895
`
`5
`
`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
`
`10
`
`known in the art.
`
`In certain embodiments of the invention, the 5-azacytidine 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 antifriction agent such as for
`
`15
`
`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 arable, gelatin, talc, titanium dioxide, or with a lacquer
`
`dissolved in the volatile organic solvent or mixture of solvents. To this coating, various dyes
`
`20 may be added in order to distinguish among tablets with different active compounds or with
`
`different amounts of the active compound present.
`
`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
`
`25
`
`powder of the 5-azacytidine polymorph in combination with a solid, pulverulent carrier, such
`
`as, for example, lactose, saccharose, sorbitol, mannitol, potato starch, corn starch,
`
`amylopectin, cellulose derivatives, or gelatin.
`
`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
`
`30
`
`particle size, and a binding agent is homogenized and suspended in a suitable solvent. The 5-
`
`azacytidine 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
`
`35
`
`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,
`
`- 1 3-
`
`Apotex v. Cellgene - IPR2023-00512
`Petitioner Apotex Exhibit 1004-0015
`
`

`

`WO 2004/082619
`
`PCT7US2004/007895
`
`5
`
`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.
`
`In the event that the above formulations are to be used for parenteral administration, such a
`
`formulation typicalty comprises sterile, aqueous and non-aqueous injection solutions
`
`10
`
`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-oxidants,
`
`buffers, bacteriostats, and solute; which render the formulation isotonic with the blood of the
`
`intended recipient. 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
`
`15
`
`example, sealed ampules and vials. Extemporaneous injection solutions and suspensions may
`
`be prepared from sterile powders, granules, and tablets of the kind previously described.
`
`Liquid preparations for oral administration are prepared 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, glycerol, and propylene glycol. If desired, such liquid
`
`20
`
`preparations contain coloring agents, flavoring agents, and saccharin. Thickening agents such
`
`as carboxymethylcellulose may also be used.
`
`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
`
`25
`
`about 5 to about 200 mg, more usually about 100 mg of the 5-azacytidine form(s). The term
`
`"unit dosage form" refers to physically discrete units suitable as unitary dosages for human
`
`subjects/patients or other mammals, each unit containing a predetermined quantity of the 5-
`
`azacytidine polymorph calculated to produce the desired therapeutic effect, in association with
`
`preferably, at least one pharmaceutically acceptable carrier, diluent, or excipient.
`
`30
`
`The following examples are provided for illustrative purposes only, and are not to be
`
`construed as limiting the scope of the claims in any way.
`
`14
`
`Apotex v. Cellgene - IPR2023-00512
`Petitioner Apotex Exhibit 1004-0016
`
`

`

`WO 2004/082619
`
`5 Examples
`
`PCT7US2004/007895
`
`Example 1: Prior Art Procedure for Synthesis of 5-azacytidine Drug Substance
`
`Using commercially available 5-azacytosine (1) and 1,2,3,5-Tetra-O- p-acetyl-
`
`ribofuranose (2) (RTA), 5-azacytidine (3) may be synthesized according to the pathway
`
`NH2
`
`NHSKCHsJj
`
`HN(Si(CH3)3),
`
`(NH2)2S04, Heat
`
`(H3C)3SiO
`
`N
`
`H O'
`
`" N'
`
`(1)
`
`NHSKCHah
`
`N^
`
`N
`
`(H^ChSiO
`
`N
`
`below.
`
`HCL
`
`(1) SnCl4,CH3CN
`
`(2) NaOCHs, CH3OH
`
`OAc
`
`"H
`
`H I
`OAc
`
`H;
`
`I H
`OAc
`
`(2)
`
`NH2
`
`N
`
`^N
`
`;H
`
`H I
`OH
`
`H:
`
`I H
`OH
`
`(3)
`
`10
`
`The crude synthesis product is dissolved in DMSO (preheated to about 90oC), and then
`methanol is added to the DMSO solution. The co-solvent mixture is equilibrated at
`approximately -20oC to allow 5-azacytidine crystal formation. The product is collected by
`vacuum filtration and allowed to air dry.
`
`Example 2: Slow Recrystallization from DMSO/toluene
`
`15
`
`Dimethyl sulfoxide (DMSO) was used as the primary solvent to solubilize Form I of 5-
`
`azacytidine and toluene was used as the co-solvent as follows. Approximately 250 mg of 5-
`
`azacytidine was dissolved with approximately 5 mL of DMSO, preheated to approximately 90
`0C, in separate 100-mL beakers. The solids were allowed to dissolve to a clear solution.
`Approximately 45 mL of toluene, preheated to approximately 50 0C, was added to the solution
`and the resultant solution was mixed. The solution was covered and allowed to equilibrate at
`
`20
`
`ambient conditions. The product was collected by vacuum filtration as white crystals using a
`
`Buchner funnel. The collected product was allowed to air dry.
`
`Example 3: Fast Recrystallization from DMSO/methanoi DMSO/toluene, and
`
`25
`
`DMSO/chloroform
`
`- 1 5-
`
`Apotex v. Cellgene - IPR2023-00512
`Petitioner Apotex Exhibit 1004-0017
`
`

`

`WO 2004/082619
`
`PCT7US2004/007895
`
`5
`
`10
`
`Approximately 250 mg of 5-azacytidine was dissolved with approximately 5 mL of
`DMSO as the primary solvent, preheated to approximately 90 0C, in separate 100-ml beakers.
`The solids were allowed to dissolve to a clear solution. Approximately 45 mL of the selected
`co-solvent (methanol, toluene, or chloroform), preheated to approximately 50 0C, was added
`to the solution and the resultant solution was mixed. The solution was covered and placed in a
`freezer to equilibrate at approximately -20oC to allow crystal formation. Solutions were
`removed from the freezer after crystal formation.
`
`The product from the methanol and toluene solutions was collected by vacuum filtration
`
`using a Buchner funnel. The resulting white crystalline product was allowed to air dry.
`
`The chloroform product was too fine to be collected by vacuum filtration. Most of the
`
`15
`
`solvent was carefully decanted from the chloroform solution and the solvent from the resultant
`
`slurry was allowed to evaporate at ambient temperature to dryness. The chloroform solution
`
`evaporated to a white product. Note that fast recrystallization using the DMSO/methanol co-
`
`solvent system has typically been used to prepare 5-azacytidine drug substance in the prior art
`
`(see the last step of the procedure provided in Example 1).