1111111111111111 IIIIII IIIII 1111111111 11111 lllll 111111111111111 11111 111111111111111 11111111
`US 20100298253Al
`
`c19) United States
`c12) Patent Application Publication
`Ionescu et al.
`
`c10) Pub. No.: US 2010/0298253 Al
`Nov. 25, 2010
`(43) Pub. Date:
`
`(54) PHARMACEUTICAL COMPOSITIONS
`COMPRISING FORMS OF 5-AZACYTIDINE
`
`(76)
`
`Inventors:
`
`Dumitru Ionescu, Ann Arbor, MI
`(US); Peter Blumbergs, Royal
`Oak, MI (US); Gary L. Silvey,
`Overland Park, KS (US)
`
`Correspondence Address:
`JONES DAY
`222 E. 41ST. STREET
`NEW YORK, NY 10017 (US)
`
`(21) Appl. No.:
`
`12/787,214
`
`(22) Filed:
`
`May 25, 2010
`
`Related U.S. Application Data
`
`(60) Continuation of application No. 11/458,365, filed on
`Jul. 18, 2006, now Pat. No. 7,772,199, which is a
`continuation of application No. 11/052,615, filed on
`Feb. 7, 2005, now Pat. No. 7,078,518, which is a divi(cid:173)
`sion of application No. 10/390,578, filed on Mar. 17,
`2003, now Pat. No. 6,887,855.
`
`Publication Classification
`
`(51)
`
`Int. Cl.
`A61K 311706
`(2006.01)
`C07H 19112
`(2006.01)
`A61P 9/00
`(2006.01)
`(52) U.S. Cl. .......................................... 514/43; 536/28.3
`ABSTRACT
`(57)
`
`The invention provides novel polymorphic and pseudopoly(cid:173)
`morphic crystalline forms of 5-azacytidine, along with meth(cid:173)
`ods for preparing said forms, wherein 5-azacytidine is repre(cid:173)
`sented by the formula:
`
`H
`
`H
`
`NH2
`NAN
`HO!AlNAO
`OH
`OH
`
`H
`
`H
`
`The invention also includes pharmaceutical compositions
`comprising said forms.
`
`CELGENE 2039
`APOTEX v. CELGENE
`IPR2023-00512
`
`

`

`Patent Application Publication Nov. 25, 2010 Sheet 1 of 8
`
`US 2010/0298253 Al
`
`(I)
`.....
`.c
`
`olLO'l£
`069£'0£
`oLLl'6t- l6~'6l
`
`ol8l'Ol---===::::
`o6t0'6 l oll9'8 l
`
`o0ff9 l---=:::::::
`o66£'t l----===.
`
`otl0'£ l--===~~
`
`ol8l'll
`
`I,...
`(I)
`'"O
`3
`
`0 a.. e I
`
`X
`
`0
`0
`0
`0
`0
`0
`0 0 0 0 0 0
`LOOLOOLOO
`LO
`LO
`..,_
`..,_
`,.-:,
`,.-:,
`
`g g g g
`
`LOO LOO
`N
`N
`
`0
`
`0
`0
`LO
`
`,...,
`
`LO
`
`0 ,...,
`
`LO
`
`0 .....
`
`

`

`Patent Application Publication Nov. 25, 2010 Sheet 2 of 8
`
`US 2010/0298253 Al
`
`l=i'
`"'O
`C:
`
`0 --c:
`
`en o
`e:..-:.
`'- 0 o·-
`1..t... -g
`Q) a::::
`ts
`_g~
`a.. ::::,
`"'O C..>
`Q) ..........
`
`Cl)
`
`-~ (I)
`::E Q)
`- "'5i
`~~
`:§CD
`~N
`0
`0
`......
`
`~~ C:
`'c5°E
`c: e
`~a..
`:i:::: Q)
`0
`'(cid:173)
`a.. 0
`E
`C:
`.Q Q)
`...... .s::.
`0
`...... e ..c:
`:::::~
`631:
`'- "'O
`CL> Q,)
`-o-
`31: ~
`0 O
`
`a.. .....J e I
`
`X
`
`LO
`I")
`
`0
`I")
`
`0 ..--
`
`N .
`(!)
`LL
`
`o H.TOZ--_,,,====-'
`o£60'6 t --o=L::::::L9:::;.8::;:=t ::::::::::-
`* \>8v9 'H
`otl9 9t --======
`ol9f v l ---===:::(cid:173)
`
`zgo·r t*o89vT :.-~l =~~=-
`
`ovvl'Zl--===:::..
`
`0
`
`0
`0
`0
`LO
`
`0
`0
`LO
`-.;:i-
`
`0
`0
`2
`...,
`
`0
`0
`LO
`I")
`
`0
`0
`0
`I")
`
`0
`0
`LO
`N
`
`0
`0
`0
`N
`
`0
`0
`LO
`...-
`
`0
`0
`0
`
`0
`
`0
`0
`I,{")
`
`

`

`Patent Application Publication
`
`Nov. 25, 2010 Sheet 3 of 8
`
`US 2010/0298253 Al
`
`Cl>
`.&::.
`
`.....
`.....
`.&::.
`"j:
`
`o£t9'lt
`oL~l'6£ --==~
`
`-0
`Cl> a;
`.Cl o.---...
`_J C
`~-Q
`I=:! t,
`E :.a
`,._ 0
`0 a:::
`LL. ~
`(1.):::.:::
`·=:::,
`;g~
`·u en
`--~
`0
`Cl>
`~ O'I
`
`0
`C (J)
`,._ N
`Cl> .....
`:::t: C
`0
`Cl>
`0.. C c·E
`.Q e
`0 a.. e Cl>
`.... ,._
`: : 0 OE
`,._
`Cl>
`-0
`31:
`0 a..
`>-e
`
`I
`X
`
`Cl>
`
`0 ..... M
`i:!::
`(.9
`I
`0
`31: LL
`I-
`
`■
`
`oltf8Z -~=== ========-:::::::::
`
`o~0£'9lo -;:£;:;tL:;.~l;-=~2
`o£9£'tl
`
`o9tt.Ll ----======:::'.
`
`08£ l'~ l ---===='.
`o680'£l ---~====-=:::
`0£86' l l ----===~
`
`099~'9
`
`0
`0
`I.(')
`I")
`
`C)
`0
`0
`I")
`
`0
`0
`I.(')
`N
`
`0
`C)
`I.(')
`
`0
`0
`0
`N
`Al!SUBlUI
`
`0
`C)
`0
`
`0
`0
`LO
`
`0
`
`0
`I")
`
`I.(')
`N
`
`0
`N
`
`I.(')
`
`0 .....
`
`I.(')
`
`

`

`Patent Application Publication Nov. 25, 2010 Sheet 4 of 8
`
`US 2010/0298253 Al
`
`Q)
`.....
`..c
`
`~
`
`Q) -a
`31=
`0 a..
`~
`I
`X
`
`o6gQ'g£
`
`o690"l£
`oU9'0£
`o£lL'8l oVll'O£
`oag·gz ol66'Ll
`o006'9Z
`o9vr·gz
`o890'tl
`9t9TZ
`
`o60L'll
`ol~WZl
`o6ZL'Olo60flZ
`olff.'Ol
`
`0£66'9 l
`ort6·g1
`
`oOLO'vl
`
`olLg'l l
`
`o990'8l
`
`o£9~tll
`
`ov0fg
`
`LO
`I"")
`
`C>
`I"")
`
`LO
`N
`
`...... ~ Q)
`0
`.
`..c
`I- (9
`I
`C> 0
`N ~ LL
`
`LO
`
`C> -
`
`LO
`
`C>
`C>
`C>
`I"")
`
`C>
`C>
`LO
`N
`
`8 C>
`
`N
`
`8
`LO
`..-
`
`C>
`C>
`C>
`
`.....
`
`C>
`C>
`LO
`
`0
`
`

`

`Patent Application Publication Nov. 25, 2010 Sheet 5 of 8
`
`US 2010/0298253 Al
`
`(1)
`
`..c ...
`..c:
`=-= 3
`
`"'O
`(1)
`(1)
`
`..0 o....--..
`....J C:
`0 >~
`E =o
`L.. 0
`oO:::
`
`(1)
`
`~~
`.s::::,
`;gs
`'<3 Cl)
`0
`(1)
`~en
`0~
`c: CD
`L..N
`2 ...
`-
`C:
`0
`(1)
`a.. C:
`c: .E
`0
`0
`· - L..
`0 a..
`e (1)
`!"=o
`-
`L..
`OE
`L..
`(1)
`"'O
`3
`0 a..
`>.. e
`I
`X
`
`o££l'l£ofl6'Q£
`o l t0'6l ofilfll
`vll'Ll o69t'9l
`
`0
`
`o6Ll'£l
`o0L6' ll
`
`a8l9'll
`
`o6t9'8l
`
`o699'tl
`
`o6l£'0l 6l8'6I
`ol0l'6l 0
`o9L6'Ll
`
`o999'9l
`
`olt9'9l
`a8t9'tl
`.
`oLtL £~gff£l
`o l9£'l l
`o8l0'l l
`
`0
`C>
`C>
`
`,.,.,
`
`C>
`0
`l.O
`N
`
`C>
`
`8
`
`N
`
`0
`0
`0
`C>
`0
`l.O
`...-
`Al!SUalUI
`
`0
`C>
`LO
`
`0
`
`0
`~
`
`l.O
`
`,.,.,
`
`0
`
`,.,.,
`
`l.O
`N
`
`... L!)
`.
`0
`(1)
`I=
`(.!J
`I
`0
`3 u..
`0
`N I-
`
`l.O
`...-
`
`0
`
`l.O
`
`

`

`Patent Application Publication Nov. 25, 2010 Sheet 6 of 8
`
`US 2010/0298253 Al
`
`QJ
`..c
`......
`..c
`......
`'ii
`""O
`QJ
`QJ
`..a
`_gc
`0 5=:g
`E~
`a a:::
`Lt... ~
`-:::.:::
`QJ .s ::,
`:e..._.,
`
`""Ou
`
`0
`Cl)
`C, <1>
`~~
`
`-~
`
`o8gl"L£
`o6LQ'g£
`o0l8'££
`
`o8gfor
`
`0Btg·5z
`o6tg·gz
`
`o0ll'Ll
`0899"9
`
`otgo·tz
`
`oar;· t z
`o80l'll o0l6'0l
`
`o0L6'l
`o£l9"ll
`
`o6l6'8 l
`
`ol08Tl
`
`o£96'Zl
`o£rg·zt
`
`0
`'q"
`
`LO
`l'0
`
`0
`l'0
`
`LO
`N
`
`0
`N
`
`LO
`
`.....
`
`0
`
`LO
`
`C,
`<1>
`
`...... (0
`~ .
`(!)
`I
`0
`:II: LL
`t-
`
`0
`0
`LO
`
`.....
`
`0
`0
`0
`
`.....
`
`0
`0
`LO
`
`AlJSU.~lUI
`
`0
`
`0
`E~
`QJ ......
`::t:: C
`C, <1>
`0... C c.E
`·- ....
`0
`0
`0 a..
`e <1>
`OE
`....
`<1>
`""O
`:II:
`0 a..
`>-
`e
`
`- .... ~o
`
`I
`X
`
`

`

`Patent Application Publication Nov. 25, 2010 Sheet 7 of 8
`
`US 2010/0298253 Al
`
`0
`-.:I"
`
`LO
`I"')
`
`0
`I"')
`
`LO
`N
`
`0
`-+-'
`Q)
`
`.=
`
`I
`0
`0
`3:
`N I-
`
`LO
`..--
`
`0
`
`r-,,..
`.
`(.9
`u..
`
`-
`
`"'O
`C: o .........
`C:
`0
`Cl) :;:;
`E.!2
`L... "O
`0
`0
`l.J.. a:::
`Q) ~
`~~
`.s::.
`::,
`a.. u
`" ' 0 -
`Q) Cl)
`-~ Q)
`:::E O'>
`Q>°~
`£co
`32N
`·u _.
`-+-'
`0 C:
`~-~
`._E
`0
`0
`L...
`ca..
`
`L...
`Q) Q)
`-+-' L...
`-+-' 0
`d!.E
`C Q)
`o.s::.
`:;:; -+-'
`(.) .s::.
`
`e~
`
`:::::: 3:
`ci"'O
`L... Q)
`Q) Q)
`"'O ..0
`3: 0
`0 _J
`a..
`>-s!
`e
`I
`X
`
`o<;tt ·a
`
`oZ8~t6Z
`o96Z'6Z
`
`*ogzg·gz
`
`ot88TZ
`*o<;9£"ll oc;c;n
`069 ·oz
`oOOl"Ol
`oWl"6l
`o6t9'8l
`o68t'9l
`
`o8tOT r8 l ft l
`*oB<;L"ll
`o80Z'Zl
`
`*
`
`*o6LC<;
`
`o6t0"£l
`
`s!
`E
`&
`..s
`~
`
`(.)
`C:
`:.,::::;
`en
`:.s
`en
`cu
`.5
`~
`:.s
`.5
`....
`
`(.)
`
`0
`0
`LO
`I"')
`
`0
`0
`0
`I"')
`
`0
`0
`LO
`N
`
`0
`0
`0
`N
`
`0
`0
`LO
`
`0
`0
`0
`
`0
`0
`LO
`
`0
`
`Al!SU8lUI
`
`

`

`Patent Application Publication Nov. 25, 2010 Sheet 8 of 8
`
`US 2010/0298253 Al
`
`0
`LL.
`
`Cl)
`C:
`::,
`~ (.)
`·u en
`-+-'
`0
`Cl)
`~0-,
`
`Cl)
`.r=
`-+-'
`.r=
`-+-'
`·3:
`"'O
`Cl)
`Q)
`..0
`..9---
`C
`!s! :g
`- 0
`"'O E~
`ts -~
`·---
`-~
`
`0 (D
`EN
`
`OJ -+-'
`~ C:
`0
`Cl)
`a..-~
`CE
`oe
`:.::;a..
`0
`0 ~
`'--
`~ 0
`Cl E
`'--Cl)
`"'O
`~
`0
`a..
`>.. e
`I
`X
`
`0
`~
`
`a90£'8£
`
`aVLO'~£
`
`a8ffl£
`
`o£6l.6l
`
`a0vt£l
`
`olgg·oz
`aS::tz·oz
`
`oga·Ll
`a6t8·gl
`
`a8g£"£l
`
`0099·01
`
`o66g·g
`
`LO
`I"')
`
`0
`I"')
`
`1.0
`N
`
`0
`N
`
`1.0
`
`0 -
`
`ot<;~·<;z
`
`aOLL'll
`
`a6v9'Zl
`
`a009'll
`
`0
`0
`LO
`I"')
`
`0
`0
`0
`I"')
`
`0
`0
`LO
`N
`
`C)
`0
`0
`N
`
`0
`0
`LO
`
`i\l!SU9lUI
`
`0
`
`C)
`C)
`LO
`
`C)
`C)
`0
`
`-
`
`0
`Cl)
`
`-+-' co
`I=
`(.9
`I
`0
`.!
`LL
`
`■
`
`

`

`US 2010/0298253 Al
`
`Nov. 25, 2010
`
`1
`
`PHARMACEUTICAL COMPOSITIONS
`COMPRISING 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-Azacyti(cid:173)
`dine", 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 of5-azacytidine
`(also known as azacitidine and 4-amino-l-~-D-ribofurano(cid:173)
`syl-S-triazin-2(1H)-one). 5-azacytidine may be used in the
`treatment of disease, including the treatment of myelodys(cid:173)
`plastic 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, interfa(cid:173)
`cial 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 phar(cid:173)
`maceutical 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 Modern 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 clini(cid:173)
`cal trials for the treatment of myelodysplastic syndromes
`(MDS). See Kornblith 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 C 8H 12N4 0 5 , a molecular weight of 244.20 and a
`structure of:
`
`NH2 NAN
`
`HO~ lNAO
`
`H
`
`H
`
`OH
`
`H
`
`H
`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
`exists in at least eight different polymorphic and pseudopoly(cid:173)
`morphic 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 each have characteristic X-ray power diffraction
`(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 Form IV, Form V, or Form
`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).
`
`DETAILED DESCRIPTION OF THE PREFERRED
`EMBODIMENTS
`5-azacytidine Crystalline Forms I-VIII
`[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.
`
`

`

`US 2010/0298253 Al
`
`Nov. 25, 2010
`
`2
`
`[0017] Form I
`[0018] A single sample of the 5-azacytidine drug substance
`was synthesized from 5-azacytosine and 1,2,3,5,-Tetra-O(cid:173)
`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 product 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 intensities
`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)
`
`20Angle ( 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 ana(cid:173)
`lyzed by XRPD. The retained drug substance samples com(cid:173)
`prised 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
`22.3° 28. Table 2 provides the most prominent 28 angles,
`d-spacing and relative intensities for this mixed phase.
`
`TABLE2
`
`5-azacytidine, Mixed Phase Forms I and II - the most prominent
`28 angles cl-spacing and relative intensities (Cu Ka radiation)
`
`20Angle ( 0
`
`)
`
`cl-spacing (A)
`
`Relative Intensity
`
`12.244
`13.082
`13.458*
`
`7.223
`6.762
`6.574
`
`34.8
`37.0
`29.2
`
`TABLE 2-continued
`
`5-azacytidine, Mixed Phase Forms I and II - the most prominent
`28 angles cl-spacing and relative intensities (Cu Ka radiation)
`
`20Angle ( 0
`
`)
`
`cl-spacing (A)
`
`Relative Intensity
`
`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
`
`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
`
`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-azacytidine
`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 pre(cid:173)
`sented 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 material. The XRPD
`powder pattern for Form III is distinctly different from that of
`all of the other forms of 5-azacytidine.
`
`TABLE3
`
`5-azacytidine, Form III - the most prominent 20 angles, cl-spacing
`and relative intensities (Cu Ka radiation)
`
`20Angle ( 0
`
`)
`
`cl-spacing (A)
`
`Relative Intensity
`
`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
`33.536
`
`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
`2.670
`
`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
`15.1
`
`

`

`US 2010/0298253 Al
`
`Nov. 25, 2010
`
`3
`
`TABLE 3-continued
`
`5-azacyticline, Form III - the most prominent 20 angles, cl-spacing
`and relative intensities (Cu Ka radiation)
`
`20Angle ( 0
`
`)
`
`cl-spacing (A)
`
`Relative Intensity
`
`36.371
`39.157
`41.643
`
`2.468
`2.299
`2.167
`
`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 crystallineformof5-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 sys(cid:173)
`tem (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 rela(cid:173)
`tive intensities for this crystalline material. The XRPD pow(cid:173)
`der pattern for Form IV is distinctly different from that of any
`other form.
`
`TABLE4
`
`5-azacytidine Form IV - the most prominent 20 angles, cl-spacing
`and relative intensities (Cu Ka radiation)
`
`20Angle ( 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 in
`Example 6.
`[0032] Form V
`[0033] Form Vis a novel crystalline form of 5-azacytidine.
`Form V was recovered by fast recrystallization of 5-azacyti(cid:173)
`dine 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 intensities for
`
`this crystalline material. The XRPD powder pattern for Form
`V is distinctly different from that of any other form.
`
`TABLES
`
`5-azacytidine Form V - the most prominent 28 angles, cl-spacing
`and relative intensities (Cu Ka radiation)
`
`20Angle ( 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 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 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.
`
`TABLE6
`
`5-azacyticline Form VI - the most prominent 20 angles, cl-spacing
`and relative intensities (Cu Ka radiation)
`
`20Angle ( 0
`
`)
`
`cl-spacing (A)
`
`Relative Intensity
`
`12.533
`12.963
`13.801
`18.929
`20.920
`21.108
`21.527
`22.623
`22.970
`24.054
`26.668
`27.210
`28.519
`29.548
`
`7.057
`6.824
`6.411
`4.6843
`4.243
`4.205
`4.125
`3.922
`3.869
`3.697
`3.340
`3.275
`3.127
`3.021
`
`10.1
`10.2
`100.0
`10.0
`34.2
`49.4
`47.0
`10.7
`13.8
`77.8
`23.0
`33.7
`12.9
`27.2
`
`

`

`US 2010/0298253 Al
`
`Nov. 25, 2010
`
`4
`
`TABLE 6-continued
`
`TABLES
`
`5-azacytidine Form VI - the most prominent 20 angles, cl-spacing
`and relative intensities (Cu Ka radiation)
`
`5-azacytidine, Form VIII - the most prominent 20 angles,
`cl-spacing and relative intensities (Cu Ka radiation)
`
`20Angle ( 0
`
`)
`
`cl-spacing (A)
`
`Relative Intensity
`
`20Angle ( 0
`
`)
`
`cl-spacing (A)
`
`Relative Intensity
`
`30.458
`33.810
`35.079
`37.528
`
`2.932
`2.649
`2.556
`2.411
`
`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 distinctive 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 20 angles,
`cl-spacing and relative intensities (Cu Ka radiation)
`
`20Angle ( 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
`
`[0040] Thermal analysis of Form VII is presented in
`Example 6
`[0041] Form VIII
`[0042] Form VIII is a novel crystalline form of 5-azacyti(cid:173)
`dine. Form VIII was recovered by recrystallizing 5-azacyti(cid:173)
`dine 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.
`
`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 propylene
`glycol, polyethylene glycol and DMSO. See Example 8.
`[0045] Pharmaceutical Formulations
`[0046] 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 excipi(cid:173)
`ent.
`[0047] 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 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 cap(cid:173)
`sules), suppositories, sterile injectable solutions, and sterile
`packaged powders.
`[0048] Examples of suitable excipients include, but are not
`limited to, starches, gum arabic, calcium silicate, microcrys(cid:173)
`talline 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; emulsi(cid:173)
`fying 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, tartrate, 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 composi(cid:173)
`tions of the invention can be formulated so as to provide
`
`

`

`US 2010/0298253 Al
`
`Nov. 25, 2010
`
`5
`
`quick, sustained, controlled, or delayed release of the drug
`substance after administration to the patient by employing
`procedures well known in the art.
`[0049]
`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 antifriction
`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 diox(cid:173)
`ide, 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.
`[0050] Soft gelatin capsules may be prepared in which cap(cid:173)
`sules contain a mixture of the 5-azacytidine form(s) and veg(cid:173)
`etable 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, pulverulent carrier,
`such as, for example, lactose, saccharose, sorbitol, mannitol,
`potato starch, corn starch, amylopectin, cellulose derivatives,
`or gelatin.
`[0051] 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 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 uni(cid:173)
`form suspension. The moistening typically causes the par(cid:173)
`ticles 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 mix(cid:173)
`ture 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.
`[0052]
`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 recipient.
`Aqueous and non-aqueous suspensions may include sus(cid:173)
`pending agents and thickening agents. The formulations may
`be present in unit-dose or multi-dose containers, for example,
`sealed ampules and vials. Extemporaneous injection solu(cid:173)
`tions and suspensions may be prepared from sterile powders,
`granules, and tablets of the kind previously described.
`[0053] 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 poly(cid:173)
`morph(s), sugar, and a mixture of ethanol, water, glycerol,
`and propylene glycol. If desired, such liquid preparations
`
`contain coloring agents, flavoring agents, and saccharin.
`Thickening agents such as carboxymethylcellulose may also
`be used.
`[0054] 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-