(12) United States Patent
`Groman et al.
`
`(10) Patent N0.:
`(45) Date of Patent:
`
`US 7,871,597 B2
`*Jan. 18, 2011
`
`US007871597B2
`
`POLYOL AND POLYETHER IRON OXIDE
`COMPLEXES AS PHARMACOLOGICAL
`AND/OR MRI CONTRAST AGENTS
`
`6,165,378 A 12/2000 Maruno et al. ......... .. 252/6253
`
`FOREIGN PATENT DOCUMENTS
`
`Inventors: Ernest V. Groman, Brookline, MA
`(US); Kenneth G. Paul, Holliston, MA
`(US); Timothy B. Frigo, Waltham, MA
`(US); Howard Bengele, Canton, MA
`(US); Jerome M. Lewis, Newton, MA
`(Us)
`
`(73)
`
`Assignee:
`
`AMAG P harmaceuticals, Inc.,
`Lexington, MA (US)
`
`Notice:
`
`Subject to any disclaimer, the term of this
`patent is extended or adjusted under 35
`USC 154(b) by 933 days.
`
`This patent is subject to a terminal dis
`claimer.
`
`(54)
`
`(75)
`
`(21)
`
`(22)
`
`(65)
`
`(63)
`
`(60)
`
`(51)
`
`(52)
`
`(58)
`
`(56)
`
`EP
`EP
`EP
`W0
`W0
`W0
`
`8/1987
`0 230 768 B1
`10/1991
`0 450 092 B1
`0450092 B1 * 8/1996
`W0 91 09678
`7/1991
`W0 96 09840
`4/1996
`W0 00 30657
`6/2000
`
`OTHER PUBLICATIONS
`
`Bulte et al. (Short-vs. Long-Circulating Magnetoliposomes as Bone
`Marrow-Seeking MR Contrast Agents, Journal of Magnetic Reso
`nance Imaging, Feb. 1999, vol. 9, pp. 329-335).*
`Briseid, G., et al., “Dextran-Induced Anaphylactoid Reaction in Man:
`Altered Reactivity of High Molecular Weight Kininogen,” Acta
`Pharmcol. Et toxicol., 1980, 47:119-126.
`Grimm, Jan, et al., “Characterization of Ultrasmall, Paramagnetic
`Magnetite Particles as Superparamagnetic Contrast Agents in MRI,”
`Database Chemabs [Online]: Invest. Radiol, 2000, vol. 35(9), pp.
`553-556.
`Hanna, C.H., et al., “Effect of Ether and Barbiturate Anesthesia on the
`Reaction of Rats to Dextran and of Dogs to Polyvinylpyrrolidone,”
`Am. J'. Physiol. 1957, 191:615-620.
`Hasegawa, et al., “Biological Behavior of Dextran-Iron Oxide Mag
`netic Fluid Injected Intravenously in Rats,” Japan J'. Appl. Phys.,
`1998, vol. 37:1029-1032.
`Hedin, H., et al., “Prevention of Dextran Anaphylaxis,” Int. Arch.
`Allergy and Immunol, 1997: 1 13:358-359.
`Jue, C.K., et al., “Determination of Reducing Sugars in the Nanomole
`Range With Tetrazolium Blue,” J'. Biochem. Biophys. Methods, 1985,
`1 1: 109-1 15.
`Kitchen, R., “Total Polysaccharide Test and Quantitative Method for
`Dextran,” Proc. Sugar Process. Res. Conf, 1983, 232-247.
`Kumar, K., “Iron Assay and Size Exclusion High Performance Liquid
`Chromatography of Ferritin and Magnetoferritin,” J'. Liq. Chrom.
`Rel. Technol., 1997, 20, 3351-3364.
`Squire, J.R., et al., “Dextran, Its Properties and Use in Medicine”
`Charles C. Thomas, Spring?eld, IL, 1955.
`Vorhees, A.B., et al., “Reactions of Albino Rats to Injections of
`Dextran,” Proc. Soc. Exp. Biol. Med., 1951, 76:254.
`Wooding, et al., “Proteins and Carbohydrates as Alternative
`Surfactants for the Preparation of Stable Magnetic Fluids,” IEEE
`Transactions on Magnetics,vol. 24,No. 2, Mar. 1998,pp. 1650-1652.
`
`* cited by examiner
`
`Primary ExamineriSreeni Padmanabhan
`Assistant ExamineriLayla Soroush
`(74) Attorney, Agent, or FirmiSunstein Kann Murphy &
`Timbers LLP
`
`(57)
`
`ABSTRACT
`
`Pharmacological compositions, and methods for administra
`tion, of the type employing an iron oxide complex With a
`polyol or polyether. The methods of administration may com
`prise parenteral administration of an effective dose of the
`complex formulated in a biocompatible liquid delivered at a
`rate of from about 1 mL/sec to less than 1 mL/min and
`Wherein upon administration the complex provides minimal
`detectable free iron in a subject, and minimal incidence of
`anaphylaxis. The pharmacological compositions are of the
`type employing a polyol or polyether iron oxide complex,
`Which, upon parenteral administration to a subject, are sub
`stantially immunosilent, provide minimal anaphylaxis and
`minimal free iron, and undergo minimal dissolution in vivo.
`
`8 Claims, 13 Drawing Sheets
`
`Appl. No.:
`
`10/410,52 7
`
`Filed:
`
`Apr. 9, 2003
`
`Prior Publication Data
`
`US 2003/0232084 A1
`
`Dec. 18, 2003
`
`Related US. Application Data
`Continuation-in-part of application No. 09/521,264,
`?led on Mar. 8, 2000, noW Pat. No. 6,599,498.
`Provisional application No. 60/128,579, ?led on Apr.
`9, 1999.
`
`Int. Cl.
`(2006.01)
`A61B 5/055
`(2006.01)
`A01N 43/04
`(2006.01)
`A61K 31/715
`US. Cl. ..................... .. 424/9.3; 424/9351; 514/23;
`514/54; 514/59
`Field of Classi?cation Search ............... .. 424/646,
`424/93; 514/23, 54, 59
`See application ?le for complete search history.
`
`References Cited
`
`U.S. PATENT DOCUMENTS
`
`10/1958
`10/1958
`12/1958
`5/1959
`9/1961
`9/1964
`7/1978
`12/1979
`6/1984
`9/1988
`5/1989
`10/1991
`12/1991
`4/1992
`11/1992
`4/1993
`11/1993
`
`Novak et al. ........... .. 252/3635
`Novak et al. .
`260/209
`Berger et a1. .
`260/209
`
`Herb ........ ..
`
`Novak ...... ..
`
`260/209
`
`260/209
`
`Heckel et a1. ............. .. 260/209
`Hasegawa et al. ...... .. 252/6253
`
`Herb ........... ..
`
`424/180
`
`Molday .................... .. 424/11
`
`Groman et al. ........... .. 128/654
`Groman et al. .
`128/653
`
`Lewis et al. . . . . .
`
`. . . . .. 424/9
`
`Ullman et a1. ..
`
`252/6251
`
`Groman et al. .
`
`.... .. 424/ 9
`
`Groman et al. .
`Maruno et al. ..
`
`.... .. 424/ 9
`536/101
`
`Palmacci et al. ............. .. 424/9
`
`2,856,366
`2,856,398
`2,862,920
`2,885,393
`3,000,872
`3,151,107
`4,101,435
`4,180,567
`4,452,773
`4,770,183
`4,827,945
`5,055,288
`5,076,950
`5,102,652
`5,160,726
`5,204,457
`
`5,262,176 >>>>>>>>>>>>>>>>>
`
`Pharmacosmos, Exh. 1018, p. 1
`
`

`

`US. Patent
`
`Jan. 18, 2011
`
`Sheet 1 0f 13
`
`US 7,871,597 B2
`
`
`
`<mmésuan 2 82mm 52% am 9.6: 258m $3M.
`
`
`
`
`
`Don-Z
`
`$3.
`
`wwwh
`
`Amnoeea $3019
`
`Pharmacosmos, Exh. 1018, p. 2
`
`

`

`US. Patent
`
`Jan. 18, 2011
`
`Sheet 2 0f 13
`
`US 7,871,597 B2
`
`Pharmacosmos, Exh. 1018, p. 3
`
`

`

`US. Patent
`
`Jan. 18, 2011
`
`Sheet 3 0f 13
`
`US 7,871,597 B2
`
`
`
`Q8 8.2 8.2 8.8
`
`
`
`8.8 8.8 8%
`
`
`
`:59 >ozm3owE
`
`m .GE
`
`Pharmacosmos, Exh. 1018, p. 4
`
`

`

`US. Patent
`
`Jan. 18, 2011
`
`Sheet 4 0f 13
`
`US 7,871,597 B2
`
`
`
`
`
`QmE .53 0:039:25
`
`- - - - - I - - b
`
`2 2. 2. Nb 2 3 8 3 3 8 .w .@ _ H_
`
`I can?
`
`1 25;.
`
`
`
`I coup,
`
`I so:
`
`
`
`. 82 9.56 9.3.1.1002
`
`Pharmacosmos, Exh. 1018, p. 5
`
`

`

`U.S. Patent
`
`Jan.18,2011
`
`31f05LI.6ehS
`
`US 7,871,597 B2
`
`I|r||L||L|IIrIIrI|L|I|______I|+||L||1|I|TI|+||L|II______IlfiIlJIIJI|l7llfiIIJIll
`
`IlflIlJIlJIIl1IlflllJlIl______
`
`IIPIILIILIIIFIIPIILIII______I|r||L||L|I|rI|r||L|II
`IIIFIILIILIII.__
`
`emSmow.om?om?888IIrIILIILIIIFIIrIILIII______I|+I|4I|JIIITII+I|4___
`
`
`
`
`
`.z__\szo_Bm_,z_-5oam__>_:
`
`muEx9NuEmD;_Em§
`
`m.0_u_
`
`
`
`3388#._.O._mmmmm_n_oo
`
`
`
`
`
`Pharmacosmos, Exh. 1018, p. 6
`
`

`

`US. Patent
`
`Jan. 18, 2011
`
`Sheet 6 0f 13
`
`US 7,871,597 B2
`
`oww
`
`
`
`HHHUHHQZJE mozwaizoo $8-1:
`..... ITIIEWMSZEE H s: -rll
`
`
`
`
`
`. 00F
`
`Pharmacosmos, Exh. 1018, p. 7
`
`

`

`US. Patent
`
`Jan. 18, 2011
`
`Sheet 7 0f 13
`
`US 7,871,597 B2
`
`Pharmacosmos, Exh. 1018, p. 8
`
`

`

`US. Patent
`
`Jan. 18, 2011
`
`Sheet 8 0f 13
`
`US 7,871,597 B2
`
`mm .OE
`
`Pharmacosmos, Exh. 1018, p. 9
`
`

`

`US. Patent
`
`Jan. 18, 2011
`
`Sheet 9 0f 13
`
`US 7,871,597 B2
`
`Pharmacosmos, Exh. 1018, p. 10
`
`

`

`US. Patent
`
`Jan. 18, 2011
`
`Sheet 10 0f 13
`
`US 7,871,597 B2
`
`Pharmacosmos, Exh. 1018, p. 11
`
`

`

`US. Patent
`
`Jan. 18, 2011
`
`Sheet 11 0f 13
`
`US 7,871,597 B2
`
`FIG. 9A
`
`\\ \\
`
`\\\\\\\\
`.
`\\\\\
`
`-
`
`\ Q Q
`\
`
`Q
`
`'
`
`\\\\.
`
`Pharmacosmos, Exh. 1018, p. 12
`
`

`

`US. Patent
`
`Jan. 18, 2011
`
`Sheet 12 0f 13
`
`US 7,871,597 B2
`
`mm .QE
`
`Pharmacosmos, Exh. 1018, p. 13
`
`

`

`US. Patent
`
`Jan. 18, 2011
`
`Sheet 13 0f 13
`
`US 7,871,597 B2
`
`BLOOD CLEARANCE IN HUMANS AT 4 mg/kg
`
`0 6 1
`
`\
`\
`\
`
`4'0
`TIME (HOURS)
`FIG. 1 0A
`
`60
`
`BLOOD CLEARANCE lN HUMANS AT 4 mg/kg
`
`1
`
`1
`
`00
`42
`
`Km.
`
`O O 1
`
`80
`
`
`
`O0 64
`
`O 2
`
`O
`
`x.
`
`0 0 0 0 0 0 0 0 0
`
`/
`/ l0.
`/ 1
`
`\ -8.
`l2. 7
`0
`0
`4
`
`l8.
`0
`
`2
`
`0.
`
`l5. 0
`
`l0.
`4
`2
`TIME (HOURS)
`FIG. 1 OB
`
`// l0 JO
`
`.0
`
`Pharmacosmos, Exh. 1018, p. 14
`
`

`

`US 7,871,597 B2
`
`1
`POLYOL AND POLYETHER IRON OXIDE
`COMPLEXES AS PHARMACOLOGICAL
`AND/OR MRI CONTRAST AGENTS
`
`CROSS REFERENCE TO RELATED
`APPLICATIONS
`
`This application is a continuation-in-part application from
`US. patent application Ser. No. 09/521,264, ?led Mar. 8,
`2000 Which in turn claims the bene?t of Provisional Appli
`cation No. 60/128,579, ?led in the United States Patent and
`Trademark Of?ce on Apr. 9, 1999, both of Which are hereby
`incorporated by reference herein.
`
`TECHNICAL FIELD AND BACKGROUND ART
`
`The ?eld relates to complexes of polyols and polyethers
`With iron oxides including a reduced polysaccharide or
`derivatiZed reduced polysaccharide, and methods for admin
`istering as pharmacological and/or MRI contrast agents.
`
`20
`
`BACKGROUND
`
`Since the invention of magnetic resonance imaging (MRI),
`a parallel technology of injectable chemicals called contrast
`agents has developed. Contrast agents play an important role
`in the practice of medicine in that they help produce more
`useful MRI images for diagnostic purposes. In particular, tWo
`classes of imaging agents have been developed and adopted in
`clinical practice. These are: loW molecular Weight gado
`linium complexes such as Magnavist®; and colloidal iron
`oxides such as Feridex I.V.® and Combidex®. Neither of
`these tWo types of agents is ideal. Problems encountered With
`these agents are shoWn in Table 1, and include: expense of
`components; ine?iciency of synthesis; loss of coating during
`terminal steriliZation (autoclaving); narroW range of organ
`uptake for purposes of imaging; toxic side-effects; restriction
`of use to either ?rst pass or equilibrium do sing, and others that
`are described herein. Agents that overcome these problems,
`and that combine the properties of these tWo types of contrast
`agents, are highly desirable.
`
`TABLE 1
`
`Comparison of ideal properties of MRI contrast agents With properties of
`low molecular Weight gadolinium based contrast agents and colloidal iron
`oxides.
`
`Properties of an ideal
`contrast agent
`
`10W molecular Weight
`gadolinium
`
`colloidal iron
`oxides
`
`LoW production costs:
`ef?cient synthesis
`Autoclavable Without
`excipients
`T1 agent
`T2 agent
`Non toxic
`Imaging vascular
`compartment at early phase
`(as a bolus administration)
`and at a late stage
`(equilibrium phase)
`Multiple administration in
`single examination
`Image of multiple target
`organs
`Bolus injection
`LoW volume of injection
`Iron source for anemia
`
`Yes
`
`Yes
`
`Yes
`No
`Yes
`No
`
`No
`
`Yes
`
`Yes
`No
`No
`
`No
`
`No
`
`Sometimes
`Yes
`No
`No
`
`No
`
`Sometimes
`
`No
`No
`Yes
`
`25
`
`30
`
`35
`
`40
`
`45
`
`50
`
`55
`
`60
`
`65
`
`2
`SUMMARY OF THE INVENTION
`
`An embodiment in accordance With the presently claimed
`invention includes an improved method for administration of
`a pharmacological composition of the type employing an iron
`oxide complex With a polyol or polyether, Wherein the
`improvement comprises administering parenterally an effec
`tive dose of an iron oxide complex With a polyol or polyether,
`the complex formulated in a biocompatible liquid so that
`upon administration the complex provides minimal detect
`able free iron in a subject and minimal incidence of anaphy
`laxis, and effecting such administration at a rate substantially
`greater than 1 mL/min or alternatively, the administration
`may be at a rate of about 1 mL/ sec.
`We have found it possible to formulate complexes having
`the properties described above. Whereas prior art complexes
`of dextran and iron oxide can be made that have minimal
`detectable free iron, and other complexes of iron oxide may
`have minimal incidence of anaphylaxis, no prior art com
`plexes of iron oxide have both properties. We have surpris
`ingly found a Way of providing a complex of modi?ed polyols
`or polyethers With iron oxide that have both properties. We
`have found, for example, that a polysaccharide such as dext
`ran, When reduced and carboxyalkylated, can be complexed
`With iron oxide to produce a composition that continues (like
`dextran iron oxide) to have minimal detectable free iron in a
`subject, While (unlike dextran iron oxide) also having mini
`mal incidence of anaphylaxis.
`Another embodiment of the present invention includes an
`improved method for administration of a pharmacological
`composition of the type employing an autoclavable reduced
`carboxyalkylated polysaccharide iron oxide complex With a
`polyol or polyether, Wherein the improvement comprises
`administering parenterally an effective dose of an iron oxide
`complex, the complex formulated in a biocompatible liquid
`so that upon administration the complex provides minimal
`detectable free iron in a subject, and minimal incidence of
`anaphylaxis, and effecting such administration at a rate sub
`stantially greater than 1 mL/min or alternatively, a rate of
`about 1 mL/ sec.
`A particular embodiment of the presently claimed inven
`tion includes an improved method for administration of a
`pharmacological composition of the type employing an iron
`oxide complex With a polyol or polyether, Wherein the
`improvement comprises parenteral administration of the
`complex to provide minimal detectable free iron in a subject
`as measured by a catalytic bleomycin assay and minimal
`incidence of anaphylaxis.
`Another particular embodiment includes an improved
`method for administration of a pharmacological composition
`of the type employing an iron oxide complex With a polyol,
`for example dextran, or polyether, for example polyethylene
`glycol, Wherein the improvement further comprises
`parenteral administration of the complex to provide minimal
`dissolution of the complex in a human subject measured as a
`function of transferrin saturation in vivo.
`Still another particular embodiment provides an improved
`method for administration of a pharmacological composition
`of the type employing an iron oxide complex With a polyol or
`polyether, Wherein the improvement further comprises
`parenteral administration of the complex to provide the
`polyol or polyether complex as an immunosilent complex in
`a human subject.
`Another particular embodiment in accordance With the
`present invention includes an improved pharmacological
`composition of the type employing an iron oxide complex
`With a polyol or polyether, Wherein the improvement com
`
`Pharmacosmos, Exh. 1018, p. 15
`
`

`

`US 7,871,597 B2
`
`3
`prises formulating a polyol or polyether complexation With
`iron oxide to provide upon administration to a subject mini
`mal detectable free iron in the subject as measured by a
`catalytic bleomycin assay and minimal incidence of anaphy
`laxis.
`Yet another embodiment in accordance With the present
`invention is an improved pharmacological composition of the
`type employing an iron oxide complex With a polyol or poly
`ether, Wherein the improvement comprises formulating a
`polyol or polyether complex With the iron oxide to provide
`upon administration to a subject minimal dissolution of the
`complex in the subject, measured as a function of transferrin
`saturation in vivo.
`Other embodiments include an improved pharmacological
`composition of the type employing an iron oxide complex
`With a polyol or polyether, Wherein the improvement com
`prises formulating a polyol or polyether complexation With
`iron oxide to provide upon administration to a subject the iron
`oxide complex as an immunosilent complex in a human sub
`ject.
`Another embodiment in accordance With the present inven
`tion includes an improved method for administration of a
`pharmacological composition of the type employing an iron
`oxide complex With a polyol or polyether, Wherein the
`improvement comprises parenteral administration of an
`effective dose of the complex formulated in a biocompatible
`liquid delivered at a rate substantially greater than 1 mL/min
`and Wherein upon administration the complex provides mini
`mal detectable free iron in a subject, and minimal incidence of
`anaphylaxis; or alternatively, the complex is delivered at a
`rate of about 1 mL/sec. More particularly, the improved
`method may utilize an assay for determining minimal detect
`able free iron Wherein the assay is any assay knoWn in the art
`for measuring free iron concentration, including a BDI assay,
`atomic absorption spectroscopy, a % transferrin saturation
`assay, a % dialysis assay, and a bacterial groWth assay. Still
`more particularly, the assay for determining minimal inci
`dence of anaphylaxis is an ELISA assay.
`In other embodiments in accordance With the invention
`includes an improved method for administering a pharmaco
`logical composition of the type employing an iron oxide
`complex With a polyol or polyether, Wherein the improvement
`comprises parenteral administration of an effective dose of
`the complex formulated in a biocompatible liquid delivered at
`a rate substantially greater than 1 mL/min, or alternatively at
`about 1 mL/sec, and Wherein upon administration the com
`plex provides minimal detectable free iron in a subject and
`minimal incidence of anaphylaxis, and Wherein the free iron
`concentration is determined using a BDI assay, and is less
`than about 750 nM, or less than about 0.04 ug/mL, or less than
`about 0.1% of the effective dose of iron oxide, depending
`upon hoW the BDI-detected free iron measurement is
`reported. In alternative embodiments, the free iron concen
`tration is determined using atomic absorption spectroscopy,
`and is less than about 1 ppm or less than about 0.04 ug/mL, or
`less than about 0.1% of the effective dose of the iron oxide,
`depending upon hoW the atomic ab sorption-detected free iron
`measurement is reported; or, the free iron concentration is
`determined using a iron dialyZed % assay, and the dialyZed
`determined free iron percent is less than about 1%.
`Yet another embodiment of the present invention includes
`an improved method for administration of a pharmacological
`composition of the type employing an iron oxide complex
`With a polyol or polyether, Wherein the improvement com
`prises parenteral administration of an effective dose of the
`complex formulated in a biocompatible liquid delivered at a
`rate substantially greater than 1 mL/min, or alternatively at a
`
`20
`
`25
`
`30
`
`35
`
`40
`
`45
`
`50
`
`55
`
`60
`
`65
`
`4
`rate of about 1 mL/ sec, and Wherein upon administration the
`complex provides minimal detectable free iron in a subject
`and minimal incidence of anaphylaxis, and Wherein the
`improvement further comprises parenteral administration of
`the complex to provide minimal dissolution of the complex in
`a human subject. More particularly, in alternative embodi
`ment, the minimal dissolution of the complex is determined
`using a % transferrin saturation assay; and more particularly,
`the minimal dissolution of the complex determined by a %
`transferrin saturation assay is less than about 95% saturation
`for a total dose from about 1 mg/kg of body Weight to about 4
`mg/kg of body Weight, up to a total single dose of about 500
`mg to about 600. An alternative embodiment further com
`prises parenteral administration of the complex to provide the
`iron oxide complex as a substantially immunosilent complex
`in a human subject. In such embodiments, veri?cation of
`administration that provides a complex that is substantially
`immunosilent in a human complex may be determined by a
`guinea pig anaphylaxis test.
`Other embodiments in accordance With the present inven
`tion include an improved pharmacological composition of the
`type employing an iron oxide complex With a polyol or poly
`ether, Wherein the improvement comprises a polyol or poly
`ether iron oxide complex composition prepared at concentra
`tions of betWeen about 1 mg/kg of body Weight to about 4
`mg/kg of body Weight in a total volume of biocompatible
`liquid from about 1 mL to about 15 mL and for a total single
`dose from about 50 mg to about 600 mg, Wherein the phar
`macological composition is capable of being parenterally
`administered to a subject at a rate substantially greater than 1
`mL/min, or alternatively at a rate of about 1 mL/ sec, and
`Wherein the iron oxide complex provides upon administration
`minimal detectable free iron in the subject and minimal inci
`dence of anaphylaxis. More particularly, the improved phar
`macological composition may further comprise an iron oxide
`complex having minimal free iron concentration in the sub
`ject. Determination of minimal free iron can be measured
`using any standard assay for measuring free iron knoWn in the
`art, including a BDI assay, atomic absorption spectroscopy, a
`% transferrin saturation assay, a % dialysis assay, or a bacte
`rial groWth assay. Alternatively, the improved pharmacologi
`cal composition may further comprise an iron oxide complex
`that undergoes minimal dissolution in a human subj ect upon
`administration to the subject. Other alternatives envision that
`the improved pharmacological composition may further
`comprise an iron oxide complex that undergoes minimal dis
`solution upon administration in a human subject. Minimal
`dissolution may be determined using a % transferrin satura
`tion assay. Alternatively, the improved pharmacological com
`position may further comprise an iron oxide complex that is
`substantially immunosilent upon administration in a human
`subject, and particularly, the improved pharmacological com
`position may further comprise an iron oxide complex that is
`substantially immunosilent in a human subject as determined
`by a guinea pig anaphylaxis test.
`Yet another embodiment in accordance With the present
`invention includes a method of treating a subject With an iron
`oxide complex to a subject in need thereof, the method com
`prising parenterally administering the complex formulated in
`a pharmaceutically acceptable formulation in a biocompat
`ible liquid, effecting administration at a rate substantially
`greater than 1 mL/ min, and providing an effective dose in the
`range of about 1 mg/kg of body Weight to about 4 mg/kg of
`body Weight in a total volume of biocompatible liquid of
`betWeen about 1 mL and 15 mL so that minimal free iron and
`minimal anaphylaxis occurs. More particularly, the method
`may comprise effecting administration at a rate of betWeen
`
`Pharmacosmos, Exh. 1018, p. 16
`
`

`

`US 7,871,597 B2
`
`5
`about 180 uL/ sec and about 1 mL/min. Still more particularly,
`the administration of the iron oxide complex provides mini
`mal dissolution of the complex in the subject and may further
`provide a substantially immunosilent complex to the subject.
`More particularly, a guinea pig test may be used to determine
`that the complex administered in the above method for treat
`ing is substantially immunosilent to the subject.
`Another embodiment of the invention includes a method of
`treating a subject With an autoclavable reduced carboxyalky
`lated polyol, for example dextran, iron oxide complex having
`at least 750 but less than 1500 umole of carboxyalkyl groups
`per gram of polyol to a subject, the method comprising
`parenterally administering the complex formulated in a phar
`maceutically acceptable formulation in a biocompatible liq
`uid, effecting administration at a rate substantially greater
`than 1 mL/min, and providing an effective dose in the range of
`about 1 mg/kg of body Weight to about 4 mg/kg of body
`Weight in a total volume of biocompatible liquid of betWeen
`about 1 mL and 15 mL so that minimal free iron and minimal
`anaphylaxis occurs.
`Yet another embodiment includes an improved pharmaco
`logical composition of the type employing an autoclavable
`carboxyalkylated polyether iron oxide complex, for example
`polyethylene glycol, Wherein the improvement comprises a
`carboxyalkylated iron oxide complex composition having at
`least 250 umole but less than 1500 mole of carboxyalkyl
`groups per gram of polyether, prepared at concentrations of
`betWeen about 1 mg/kg of body Weight to about 4 mg/kg of
`body Weight in a total volume of biocompatible liquid from
`about 1 mL to about 15 mL and for a total single dose from
`about 50 mg to about 600 mg, Wherein the pharmacological
`composition is capable of being parenterally administered to
`a subject at a rate substantially greater than 1 mL/min and
`Wherein the iron oxide complex provides upon administration
`minimal detectable free iron in the subject and minimal inci
`dence of anaphylaxis.
`Another embodiment of the invention is a method of pro
`viding an iron oxide complex for administration to a mammal
`subject, the method comprising: producing a reduced
`polysaccharide iron oxide complex, and sterilizing the com
`plex by autoclaving. In general, the reduced polysaccharide is
`a reduced polymer of glucose. An example of a reduced
`polymer of glucose is a reduced dextran. The reduced
`polysaccharide is produced through reaction of a polysaccha
`ride With a reagent selected from the group consisting of a
`borohydride salt or hydrogen in the presence of a hydrogena
`tion catalyst. In a further aspect of the method, the iron oxide
`is superparamagnetic.
`Another particular embodiment of the invention is a
`method of providing an iron oxide complex for administra
`tion to a mammalian subject, the method comprising: produc
`ing a derivatized reduced polysaccharide iron oxide complex,
`and sterilizing the complex by autoclaving. According to this
`method, producing the complex can include derivatizing a
`reduced polysaccharide by formation of, for example, ethers,
`amides, esters, and amines at the hydroxyl positions of the
`polysaccharide. In a particular embodiment, the derivative
`formed is an ether of the polysaccharide, more particularly a
`carboxyalkyl ether of the polysaccharide, and more particu
`larly, a carboxymethyl ether of the polysaccharide. Further
`according to this method, the reduced polysaccharide can be
`a reduced dextran. The derivatized, reduced polysaccharide
`can be isolated as the sodium salt and does not contain an
`infrared absorption peak in the region of 1650-1800 cm_l. In
`one aspect of the method, producing the derivatized reduced
`polysaccharide is achieved at a temperature of less than
`approximately 50° C. In another aspect of the method, pro
`
`20
`
`25
`
`30
`
`35
`
`40
`
`45
`
`50
`
`55
`
`60
`
`65
`
`6
`ducing the derivatized reduced polysaccharide is achieved at
`a temperature of less than approximately 400 C. In a further
`aspect of the method, the iron oxide is superparamagnetic.
`In yet another embodiment, the invention provides a
`method of formulating an iron oxide complex coated With a
`reduced polysaccharide. This composition is for pharmaco
`logical use and the composition has decreased toxicity in
`comparison to a formulation of an iron oxide complex coated
`With the non-reduced polysaccharide. The method of formu
`lating such an iron oxide complex comprises: producing a
`reduced polysaccharide iron oxide complex, and sterilizing
`the complex by autoclaving. The formulation provides a
`polysaccharide Which Was produced by reacting the polysac
`charide With one of a reducing agent selected from the group
`consisting of a borohydride salt or hydrogen in the presence
`of an hydrogenation catalyst, Wherein the reduced polysac
`charide iron oxide complex so made has such decreased tox
`icity. In a further aspect of the method, the iron oxide is
`superparamagnetic.
`In yet another embodiment, the invention provides a
`method of formulating an iron oxide complex coated With a
`reduced derivatized polysaccharide. This composition is for
`pharmacological use and the composition has decreased tox
`icity in comparison to a formulation of an iron oxide complex
`coated With the non-reduced derivatized polysaccharide. The
`method of formulating such an iron oxide complex com
`prises: producing a reduced derivatized polysaccharide iron
`oxide complex; and sterilizing the complex by autoclaving.
`According to this method, producing the complex can include
`derivatizing a reduced polysaccharide by carboxyalkylation,
`for example, Wherein the carboxyalkylation is a carboxym
`ethylation. Further according to this method, the reduced
`polysaccharide can be a reduced dextran. The derivatized,
`reduced polysaccharide can be isolated as the sodium salt and
`does not contain an infrared absorption peak in the region of
`1650-1800 cm“. In one aspect of the method, producing the
`derivatized reduced polysaccharide is achieved at a tempera
`ture of less than approximately 50° C. In another aspect of the
`method, producing the derivatized reduced polysaccharide is
`achieved at a temperature of less than approximately 400 C. In
`a further aspect of the method, the iron oxide is superpara
`magnetic.
`Another embodiment of the invention provides a reduced
`derivatized polysaccharide iron oxide complex With T1 and
`T2 relaxation properties to alloW contrast agent signal
`enhancement With T1 sequences and signal diminishment
`With T2 sequences. A further aspect of the embodiment is that
`the reduced derivatized polysaccharide iron oxide can be
`administered multiple times for sequential imaging in a single
`examination. Yet another aspect of the agent is that it can be
`used to image multiple organ systems including the vascular
`system, liver, spleen, bone marroW, and lymph nodes.
`Another embodiment of the invention provides a reduced
`polysaccharide iron oxide complex for use as an intravenous
`iron supplement.
`Another embodiment of the invention provides a reduced
`derivatized polysaccharide iron oxide complex for use as an
`intravenous iron supplement.
`In yet a further embodiment, the invention provides an
`improved method of administering to a mammalian subject
`an autoclaved reduced polysaccharide iron oxide complex.
`The improved method of administration comprising: injec
`tion of an autoclaved reduced polysaccharide iron oxide com
`plex in a volume of 15 mL or less. In another aspect of the
`embodiment the injected volume is injected as a bolus. In a
`further aspect of the method, the iron oxide is superparamag
`
`Pharmacosmos, Exh. 1018, p. 17
`
`

`

`US 7,871,597 B2
`
`7
`netic. In a further aspect of the embodiment the injected
`volume provides improved image quality.
`In yet a further embodiment, the invention provides an
`improved method of administering to a mammalian subject
`an autoclaved derivatiZed reduced polysaccharide iron oxide
`complex, the improved method of admini stration comprising:
`injection of an autoclaved reduced derivatiZed polysaccha
`ride iron oxide complex in a volume of 15 mL or less. In
`another aspect of the embodiment the injected volume is
`injected as a bolus. In a further aspect of the method, the iron
`oxide is superparamagnetic. In a further aspect of the embodi
`ment the injected volume provides improved image quality.
`An embodiment of the invention provides an improved
`method of administering to a mammalian subject a reduced
`polysaccharide iron complex to a mammalian subject
`Wherein the improvement comprises administration of a
`reduced polysaccharide in formulation to provide reduced
`toxicity relative to administration of a non-reduced polysac
`charide. In a further aspect of the embodiment, the iron oxide
`is superparamagnetic.
`An embodiment of the invention provides an improved
`method of administering to a mammalian subject a reduced
`derivatiZed polysaccharide iron complex in a manner that the
`composition provides reduced toxicity, Wherein the improve
`ment comprises utiliZing a reduced derivatiZed polysaccha
`ride in formulation of the composition. In a further aspect of
`the embodiment, the iron oxide is superparamagnetic.
`An embodiment of the invention provides a reduced
`polysaccharide iron oxide complex, Wherein the reduced
`polysaccharide is derivatiZed, for example, the reduced
`derivatiZed polysaccharide is a carboxyalkyl polysaccharide.
`The carboxyalkyl is selected from the group consisting of
`carboxymethyl, carboxyethyl and carboxypropyl. Further,
`the reduced polysaccharide can be a reduced dextran, for
`example, the reduced dextran can be a reduced carboxym
`ethyl dextran. A further aspect of this embodiment of the
`invention is that the level of derivatiZation of the reduced
`dextran is at least 750 umole but less than 1500 umole of
`carboxyl groups per gram of polysaccharide Wherein said
`composition has reduced toxicity relative to composition
`With respect to loWer levels of derivatiZation.
`An embodiment of the invention provides a reduced
`polysaccharide iron oxide complex, such complex being
`stable at a temperature of at least approximately 100° C. In a
`preferred embodiment, such complex is stable at a tempera
`ture of approximately 121° C. In an even more preferred
`aspect of the reduced polysaccharide iron oxide complex,
`such complex is stable at a temperature of at least 121° C. for
`a time su?icient to steriliZe the complex. In a furt