(19) United States
`(12) Patent Application Publication (10) Pub. No.: US 2012/0009121 A1
`Pomper et al.
`(43) Pub. Date:
`Jan. 12, 2012
`
`US 2012.0009121A1
`
`(54)
`
`PSMA-TARGETING COMPOUNDS AND USES
`THEREOF
`
`(75)
`
`Inventors:
`
`(73)
`
`Assignee:
`
`(21)
`(22)
`(86)
`
`Appl. No.:
`
`PCT Fled:
`
`PCT NO.:
`
`Martin Pomper, Baltimore, MD
`(US); Ronnie Charles Mease,
`Fairfax, VA (US); Ray Sangeeta,
`Ellicott City, MD (US); Ying Chen,
`Timonium, MD (US)
`The Johns Hopkins University,
`Baltimore, MD (US)
`13/257,499
`
`Mar. 19, 2010
`
`Publication Classification
`
`(51) Int. Cl.
`A6II 5L/00
`A61R 49/00
`CI2N 5/00
`C07D 3II/82
`C07D 403/06
`C07D 249/04
`C07D 213/53
`C07F 5/02
`C07F 5/00
`C07C323/22
`C07D 257/02
`C07F 15/00
`CI2O 1/02
`C07D 3II/78
`
`(2006.01)
`(2006.01)
`(2006.01)
`(2006.01)
`(2006.01)
`(2006.01)
`(2006.01)
`(2006.01)
`(2006.01)
`(2006.01)
`(2006.01)
`(2006.01)
`(2006.01)
`(2006.01)
`
`(60)
`
`S371 (c)(1),
`Sep. 19, 2011
`(2), (4) Date:
`Related U.S. Application Data
`Provisional application No. 61/161,485, filed on Mar.
`19, 2009, provisional application No. 61/161,484.
`filed on Mar. 19, 2009, provisional application No.
`61/248,067, filed on Oct. 2, 2009, provisional applica
`tion No. 61/248,934, filed on Oct. 6, 2009.
`
`(52) U.S. Cl. ........... 424/1.11: 435/29: 424/9.1; 435/325:
`549/388: 548/455; 549/382:546/331:546/13;
`556/1:562/556; 540/474:546/2: 548/255
`
`ABSTRACT
`(57)
`Prostate-specific membrane antigen (PSMA) targeting com
`pounds are described. Uses of the compounds for imaging,
`therapy, cell sorting, and tumor mapping are also described.
`
`Petitioner GE Healthcare – Ex. 1005, p. 1
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`FIG. 1
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`s
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`0=y_ºHOOËN. NË OZOH H HYr×
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`Petitioner GE Healthcare – Ex. 1005, p. 5
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`Petitioner GE Healthcare – Ex. 1005, p. 6
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`Petitioner GE Healthcare – Ex. 1005, p. 7
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`Petitioner GE Healthcare – Ex. 1005, p. 8
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`Petitioner GE Healthcare – Ex. 1005, p. 9
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`Patent Application Publication
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`Petitioner GE Healthcare – Ex. 1005, p. 10
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`Petitioner GE Healthcare – Ex. 1005, p. 11
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`DORSAL
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`WENTRAL
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`PRE-NECTION
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`Petitioner GE Healthcare – Ex. 1005, p. 12
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`Petitioner GE Healthcare – Ex. 1005, p. 13
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`Petitioner GE Healthcare – Ex. 1005, p. 14
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`Petitioner GE Healthcare – Ex. 1005, p. 15
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`Petitioner GE Healthcare – Ex. 1005, p. 16
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`Petitioner GE Healthcare – Ex. 1005, p. 17
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`Patent Application Publication
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`
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`am
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`Petitioner GE Healthcare – Ex. 1005, p. 18
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`Petitioner GE Healthcare – Ex. 1005, p. 19
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`Patent Application Publication
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`OMIN
`
`30 MIN
`
`FIG. 19
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`Petitioner GE Healthcare – Ex. 1005, p. 20
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`Petitioner GE Healthcare – Ex. 1005, p. 21
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`Patent Application Publication
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`US 2012/0009121 A1
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`
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`a
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`l
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`Petitioner GE Healthcare – Ex. 1005, p. 22
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`Patent Application Publication
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`Jan. 12, 2012 Sheet 22 of 27
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`US 2012/0009121 A1
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`VENTRAL
`
`DORSA
`
`PIPSIDE
`
`FLUSIDE
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`BLOOD
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`FIG.22
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`Petitioner GE Healthcare – Ex. 1005, p. 23
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`Patent Application Publication
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`Jan. 12, 2012 Sheet 23 of 27
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`US 2012/0009121 A1
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`900'd\'ONT
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`Petitioner GE Healthcare – Ex. 1005, p. 24
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`Patent Application Publication
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`Jan. 12, 2012 Sheet 24 of 27
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`US 2012/0009121 A1
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`*****®
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`Petitioner GE Healthcare – Ex. 1005, p. 25
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`Patent Application Publication
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`Jan. 12, 2012 Sheet 25 of 27
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`US 2012/0009121 A1
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`
`
`SPECIMEN OO1-PipZERO
`(x1,000) 250; .333.
`200
`150
`?5 100
`50
`
`SEE,
`(x1,000)
`FSCA
`
`TUBE. PipZERO1
`POPULATION
`ALLEVENTS
`
`SPECIMEN OO1-Pip 1000
`(x1,000) 250 s???
`200
`15
`&5 100
`50
`
`SPECIMEN_001-PipZERO1
`
`105
`
`:
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`HEVENTS
`1334,712
`991,055
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`55
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`9%PARENT 96TOTAL
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`SPECIMEN_001-Pip 1000
`105
`104
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`FIG.25B
`
`TUBE. Pip 1000
`POPULATION
`ALLEVENTS
`
`50 OO 150 200 250
`(x1,000)
`FSC-A
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`102 103 104 105
`FITC-A
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`HEVENTS
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`%PARENT %TOTAL
`100.0
`743
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`FIG. 25A
`
`Petitioner GE Healthcare – Ex. 1005, p. 26
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`Patent Application Publication
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`Jan. 12, 2012 Sheet 26 of 27
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`US 2012/0009121 A1
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`
`
`SPECIMEN OO1-Pip 10
`
`p3 p.
`404 105
`102 103
`FITCA
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`HEVENTS
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`SPECIMEN OO-Pip (OK-3
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`SPECIMEN OO1-Pip 10 K-3
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`FIG. 25B
`
`Petitioner GE Healthcare – Ex. 1005, p. 27
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`Patent Application Publication
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`Jan. 12, 2012 Sheet 27 of 27
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`US 2012/0009121 A1
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`FROM
`FIG.25B
`
`
`
`(X1,
`
`SPECIMEN OO-Pip 100
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`SPECIMEN OO1-Pip 100
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`POPULATION
`ALLEVENTS
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`POPULATION
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`SPECIMEN OO-Pip 100-K3
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`50 OO 150 200 250
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`
`FIG. 25C
`
`Petitioner GE Healthcare – Ex. 1005, p. 28
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`US 2012/0009121 A1
`
`Jan. 12, 2012
`
`PSMA-TARGETING COMPOUNDS AND USES
`THEREOF
`
`CROSS-REFERENCE TO RELATED
`APPLICATION
`0001. This application claims priority to U.S. Provisional
`Application Nos. 61/161,484 filed Mar. 19, 2009, 61/161,
`485, filed Mar. 19, 2009, 61/248,067, filed Oct. 2, 2009, and
`61/248,934, filed Oct. 6, 2009. The entire content of each
`Provisional Application is hereby incorporated by reference
`in their entirety.
`0002 This invention was made using U.S. Government
`support under NIH grant NIH U24 CA92871. The govern
`ment has certain rights in this invention.
`
`BACKGROUND
`0003 1. Field of the Invention
`0004. The present invention relates to prostate specific
`membrane antigen (PSMA) binding compounds, chemical
`precursors of PSMA binding compounds and imaging meth
`ods of using the compounds.
`0005 2. Background
`0006 Prostate cancer (PCa) is the most commonly diag
`nosed malignancy and the second leading cause of cancer
`related death in men in the United States (Cancer Facts &
`Figures: American Cancer Society: Atlanta, Ga., 2009). In
`2009, it is estimated that 192,000 men will be diagnosed with
`prostate cancer and 27,000 men will die of the disease. Only
`one half of tumors due to PCa are clinically localized at
`diagnosis and one half of those represent extracapsular
`spread. Localization of that spread as well as determination of
`the total body burden of PCa have important implications for
`therapy, particularly as new combination and focal therapies
`become available.
`0007. The prostate-specific membrane antigen (PSMA),
`while expressed in prostate tumor epithelium, has a curious
`property in that it is expressed in the neovasculature of many
`Solid tumors but not in that of prostate cancer (Chang et al.,
`Cancer Res., vol. 59, pp.3192-3 198, 1999; Changet al., Clin.
`Cancer Res., vol. 5, pp. 2674-2681, 1999; Gong et al., Cancer
`Metastasis Rev., vol. 18, pp. 483-490, 1999; Chang et al.,
`Mol. Urol. Vol. 3, pp. 313-320, 1999; Baccala et al., Urology,
`vol. 70, pp. 385-390, 2007: Changet al., Urology, vol. 57, pp.
`801-805, 2001 Milowsky et al., J. Clin. Oncol., vol. 25, pp.
`540-547, 2007). Because of that property, an ''' In-labeled
`monoclonal antibody to an extracellular epitope of PSMA,
`'''In-J591, was capable of identifying renal, bladder, lung,
`breast, colorectal and pancreatic tumors in a Phase I clinical
`imaging study (Milowsky et al., J. Clin. Oncol., Vol. 25, pp.
`540-547, 2007). That study validated '''In-J591 as a vascular
`targeting agent in human Subjects. Since then other reports
`have further studied PSMA expression in certain tumor types.
`Baccala et al. noted that clear cell renal cell carcinoma
`expresses significantly more PSMA in its neovasculature than
`does the papillary variety (Baccala et al., Urology, Vol. 70, pp.
`385-390, 2007). Furthermore, angiomyolipoma, a benign
`renal lesion, did not express PSMA. As an enzyme with an
`extracellular active site, PSMA represents an excellent target
`for imaging and therapy directed toward Solid tumor neovas
`culature in addition to prostate cancer itself. PSMA-based
`agents can report on the presence of this marker, which is
`increasingly recognized as an important prognostic determi
`nate in PCa (Murphy et al., Urology, vol. 51, pp. 89-97, 1998).
`
`It is also the target for a variety of new PCatherapies (Galsky
`et al., J Clin Oncol, vol. 26, pp. 2147-2154, 2008).
`0008 ProstaScintTM is an 'In-labeled monoclonal anti
`body against PSMA that is clinically available for imaging
`PCa. Radioimmunotherapy based on ProstaScintTM and
`radiolabeled variations of this antibody are fraught with simi
`lar difficulties to the use of radiolabeled antibodies for imag
`ing, including prolonged circulation times, poor target to
`nontarget tissue contrast, unpredictable biological effects and
`the occasional need for pre-targeting strategies, limiting the
`utility of these agents (Lange, P. H., Urology, vol. 57, pp.
`402-406, 2001; Haseman et al., Cancer Biother Radiopharm,
`vol. 15, pp. 131-140, 2000; Rosenthal et al., Tech Urol, vol. 7,
`pp. 27-37, 2001). Furthermore, antibodies may have less
`access to tumor than low molecular weight agents, which can
`be manipulated pharmacologically.
`0009. The development of low molecular weight radio
`therapeutic agents is much different from developing radiop
`harmaceuticals for imaging in that longer tumor residence
`times can often be important for the former.
`0010 Complete detection and eradication of primary
`tumor and metastatic foci are required to effect a cure in
`patients with cancer; however, current preoperative assess
`ment often misses Small metastatic deposits. More sensitive
`imaging techniques than computed tomography, magnetic
`resonance imaging and even positron emission tomography
`(PET), which can be used easily in the operating suite, are
`required. An old technique, recently revisited because of
`improved optics and fluorescent dye chemistry, is intraopera
`tive photodiagnosis (PDD) (Toda, Keio J. Med., vol. 57, pp.
`155-161, 2008). Fluorescein dyes have been used intraopera
`tively to identify brain tumors and verify the clarity of tumor
`margins since 1948 (Toda, Keio J.Med., vol. 57, pp. 155-161,
`2008). A recent report describes its utility in identifying brain
`metastases (Okuda et al., Minim. Invasive NeuroSurg. Vol.
`50, pp. 382-384, 2007). A long history of the use of 5-ami
`nolevulinic acid (5-ALA) for brain tumor resection is also
`evident, and its use has been associated with improvement in
`progression-free Survival (Stummer et al., Lancet Oncol. Vol.
`7, pp. 392–401, 2006). PDD can be performed easily during
`Surgery due to the lack of a need for complex imaging equip
`ment. All that is needed is a light-emitting diode to excite the
`fluorophore, which can be administered systemically or
`“painted on the tissue directly. More recent incarnations of
`PDD have used quantum dots (Arndt-Jovinet al., IEEE Trans
`Nanobioscience, 2009), and more advanced dyes, such as
`indocyanine green (ICG) (Gotoh et al., J. Surg. Oncol., 2009),
`which emit in the near-infrared (NIR) region of the spectrum,
`enabling reasonable tissue penetration of emitted (and
`detected) light. Applications have included nontargeted
`approaches, such as preoperative evaluation of the vascular
`integrity of surgical flaps or identification of nodules of hepa
`tocellular carcinoma (Matsui et al., Plast. Reconstr. Surg.
`vol. 123, pp. 125e-127e, 2009). Targeted approaches are also
`emerging, such as use of a fluorophore-conjugated anti-CEA
`antibody to identify colon or pancreatic cancer (Kaushal et
`al., J. Gastrointest. Surg., vol. 12, pp. 1938-1950, 2008), or
`the use of NIR activatable probes that emit light only when
`cleaved by a tumor-associated protease (Shethet al., Gynecol.
`Oncol., vol. 112, pp. 616-622, 2009).
`10011
`Recently, the application of “Ga-labeled peptides
`has attracted considerable interest for cancer imaging
`because of the physical characteristics of Ga-68 (Reubiet al.,
`JNucl Med, vol. 49, pp. 1735-1738, 2008). Ga-68 is available
`
`Petitioner GE Healthcare – Ex. 1005, p. 29
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`US 2012/0009121 A1
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`Jan. 12, 2012
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`from an in-houseGe?' Gagenerator (“Get 270.8 day),
`which renders it independent of an onsite cyclotron. There
`fore, Ga-based PET agents possess significant commercial
`potential and serve as a convenient alternative to cyclotron
`based isotopes for positron emission tomography (PET), Such
`as 'F or '''I. Gahas a high positron-emitting fraction (89%
`of its total decay). The maximum positron energy of “Ga
`(max. energy=1.92 MeV, mean=0.89 MeV) is higher than
`that of F (max=0.63 MeV, mean=0.25 MeV). However, a
`study of spatial resolution using Monte Carlo analysis
`revealed that under the assumption of 3 mm spatial resolution
`for most PET detectors, the full-width-at-half-maximum
`(FWHM) of F and Ga are indistinguishable in soft tissue
`(3.01 mm vs. 3.09 mm) (Sanchez-Crespo et al., Eur J. Nucl
`Med Mol Imaging, vol. 31, pp. 44-51, 2004). That finding
`implies that with the standard spatial resolution of 5 to 7 mm
`for current clinical scanners, image quality using “Ga-based
`radiotracers will likely be indistinguishable from that of F
`based agents, stimulating interest in the development of Ga
`labeled compounds for medical imaging (Sanchez-Crespo et
`al., Eur J. Nucl Med Mol Imaging, vol. 31, pp. 44-51, 2004:
`Khanet al., EurJSurg. Oncol, vol.35, pp. 561-567, 2009; Fani
`et al., Contrast Media Mol Imaging, vol. 3, pp. 67-77, 2008).
`With a physical half-life of 68 min, Ga is also matched
`nicely to the pharmacokinetics of many peptides used for
`imaging. Few Ga-labeled, mechanism-based radiotracers
`for prostate cancer have been reported previously, and none
`for PSMA. Furthermore, Ga is introduced to biomolecules
`through macrocyclic chelators, which allows possible kit for
`mulation and wide availability of the corresponding imaging
`agents.
`
`SUMMARY OF THE INVENTION
`0012. The present invention satisfies the long standing and
`unmet need for new imaging and therapeutic compounds for
`targeting prostate cancer and cancer angiogenesis. The
`present invention, in particular, provides therapeutic com
`pounds and imaging agents which differ from the prior art in
`modifications which were not previously known or Sug
`gested. Furthermore, the invention provides imaging agents
`that offer better contrast between target tissues and non-target
`tissues. The invention also provides compounds with greater
`cellular retention and low molecular weight.
`0013 Embodiments of the invention include compounds
`having the structure
`
`independently selected from hydrogen or a protecting group,
`a is 1, 2, 3, or 4, and R is each independently Hor C-C alkyl.
`00.15
`Variable r is 0 or 1. TZ is a triazole group selected
`from the group consisting of
`
`NEN - - - and
`---C-
`
`N
`
`where L is
`
`(CH2)
`
`O
`
`R5
`
`1– (CH2)
`
`X2
`
`s
`
`L is --to- O
`
`NRC(S)NR , or
`X is NRC(O) , NRC(O)NR
`NRC(O)NR
`NRC(O)O : X is
`C(O)NR ,
`NRC(S)NR , or OC(O)NR : R is H, COH, or
`CO.R, where R is a C-C alkyl, C-C2 aryl, or Ca-Ca
`alkylaryl; b is 1, 2, 3, or 4; and d is 1, 2, 3, or 4.
`(0016 Variable q is 0 or 1. W is NRC(O) , – NRC(O)
`NR NRC(S)NR
`NRC(O)O
`OC(O)NR
`OC(O) , —C(O)NR-, or -C(O)O ; R and R are
`independently H, CO.H, or COR', where R is a C-C,
`alkyl, C-C, aryl, or C-C alkylaryl, wherein if one of R
`and R is CO.H or CO.R, then the other is H; n is 1, 2, 3, 4,
`5 or 6.
`
`R
`
`N
`
`G
`
`2
`J. LR
`
`O
`
`R3
`
`(CH2)
`
`R
`
`W Tzi
`g
`
`(CH,)-Y-N
`
`Y(CH), O
`
`J. C.
`
`QOC
`
`COQ
`
`0014 wherein the subunits associated with elements p, q,
`r, and S may be in any order. Z is tetrazole or COQ; each Q is
`
`(0017 Variables is 0 or 1. Y is –C(O)-, - NRC(O) ,
`—NRC(S)— —OC(O); and m is 1, 2, 3, 4, 5, or 6.
`
`Petitioner GE Healthcare – Ex. 1005, p. 30
`
`

`

`US 2012/0009121 A1
`
`Jan. 12, 2012
`
`0018 Variable p is 0, 1, 2, or 3, and when p is 2 or 3, each
`R" may be the same or different. R' is H. C-C alkyl, C-C,
`aryl, or C-C alkylaryl.
`0019 G is a moiety selected from the group consisting of
`
`0021 2) when G is
`
`-----or, V
`
`R
`
`V.
`
`V.
`
`Fo1 SX s
`V
`R
`
`A-W
`YNH R
`
`R
`V
`N-(CH),
`A-W
`
`N
`
`,
`
`and
`
`O
`
`O
`
`HO
`
`NH2
`
`e
`
`N
`
`NS N
`
`and r is 0, then q and s are both 0 or both 1:
`0022 3) when G is
`
`O
`
`HO
`
`NH2
`
`e
`N
`NSN
`
`then p is 0 and R is H, and the structure optionally includes
`a chelated metalion.
`0023 4) when G is
`
`where Ch is a metal chelating moiety, optionally including a
`chelated metal; FG is a fluorescent dye moiety which emits in
`the visible or near infrared spectrum; one of A and A' is Ch
`and the other is FG: V and V are independently —C(O)—
`NRC(O)— —NRC(S)—, or —OC(O)—, and g is 1, 2, 3,
`4, 5, or 6. The following conditions also apply:
`(0020. 1) when G is
`
`and r is 0, then if p is 0, then one of R and R is COR, and
`the other is H; and
`0024 5) when g is
`
`A-W
`
`R
`
`V.
`ch1 NN
`V
`R
`
`, or
`
`N-(CH2)
`M
`A-V
`
`and r is 0, then q and S are both 1:
`
`H
`
`O
`
`N
`
`-- O --
`
`then r is 0.
`0025 Embodiments include compounds having the struc
`ture
`
`R1
`
`R
`
`N
`
`R3
`
`(CH2)
`
`V.
`C1
`
`R
`
`O
`
`R2
`p
`
`R
`
`Petitioner GE Healthcare – Ex. 1005, p. 31
`
`

`

`US 2012/0009121 A1
`
`Jan. 12, 2012
`
`wherein Z is tetrazole or COQ; each Q is independently
`selected from hydrogen or a protecting group, a is 1, 2, 3, or
`4, and Ris each independently H or C-C alkyl. Ch is a metal
`chelating moiety optionally including a chelated metal. W is
`NRC(O) , NRC(O)NR NRC(S)NR
`NRC(O)
`O-, - OC(O)NR - OC(O) = C(O)NR , or C(O)
`O-Y is C(O)-, - NRC(O) - NRC(S)-, - OC(O).
`Vis-C(O) - NRC(O) , NRC(S)-, or - OC(O)
`In exemplary embodiments m is 1, 2, 3, 4, 5, or 6; n is 1, 2, 3,
`4, 5 or 6; and p is 0, 1, 2, or 3, and when p is 2 or 3, each R'
`may be the same or different. R' is H, C-C alkyl, C-C,
`aryl, or C-C alkylaryl. R and R are independently H,
`CO.H, or CO.R, where R is a C-C alkyl, C-C aryl, or
`
`C-C alkylaryl, wherein when one of RandR is CO, Hor
`COR, the other is H, and when p is 0, one of R and R is
`COR, and the other is H.
`0026. Some embodiments further include a chelated
`metal. In some embodiments, the chelated metal is Tc, In, Ga.
`Y. Lu, Re, Cu, Ac, Bi, Pb, Sm, Sc, Co, Ho, Gd, Eu, Tb, or Dy.
`In some embodiments, the chelated metal an isotope, for
`example. In some embodiments, the isotope is Tc-94m,
`Tc-99m, In-111, Ga-67, Ga-68, Y-86, Y-90, Lu-177, Re-186,
`Re-188, Cu-64, Cu-67, Co-55, Co-57, Sc-47, Ac-225,
`Bi-213, Bi-212, Pb-212, Sm-153, Ho-166, or Dy-166.
`Embodiments include compounds having the structure
`
`O
`
`HO
`
`NH2
`
`e
`
`N
`NS/
`
`R3
`
`(CH2)
`
`W g
`
`R
`
`(CH2)-Y-N
`s (CH), O C
`---,
`QOC
`COQ
`
`H
`
`H
`
`optionally including a chelated metal ion. Z is tetrazole or
`COQ; each Q is independently selected from hydrogen or a
`protecting group, and a is 1,2,3, or 4. Ris each independently
`Hor C-C alkyl. Wis NRC(O) , NRC(O)NR , NRC
`(S)NR-, - NRC(O)O-, - OC(O)NR
`OC(O) ,
`C(O)NR , or - C(O)O - Y is C(O) - NRC(O) ,
`- NRC(S) —OC(O)–:
`0027. In exemplary embodiments m is 1, 2, 3, 4, 5, or 6; in
`is 1,2,3,4, 5 or 6: q is 0 or 1; and s is 0 or 1. R is H, CO.H.
`or CO.R, where R is a C-C alkyl, C-C aryl, or C-C,
`alkylaryl. Some embodiments further include a chelated
`metalion. In some embodiments, the metalion is Tc, Re, Cu,
`or Ga. In some embodiments, the metal ion is Tc-99m,
`Re-186, Re-188, Cu-64, or Ga-68. In some embodiments, the
`metalion is Tc-99m.
`0028 Embodiments include compounds having the struc
`ture
`
`R1
`
`R
`
`N
`
`V.
`FC1 n N
`
`R
`
`O
`
`p
`
`R2
`
`R3
`
`(CH2)
`
`R
`
`QOC
`
`g
`
`W (CH)-Y-N
`sy(CH2). O
`-Nulls
`
`Z
`
`CO
`2O
`
`N
`H
`
`N
`H
`
`Petitioner GE Healthcare – Ex. 1005, p. 32
`
`

`

`US 2012/0009121 A1
`
`Jan. 12, 2012
`
`where p, q, and S are in the order drawn, and q and S are either
`both 0 or both 1. Z is tetrazole or COQ; each Q is indepen
`dently selected from hydrogen or a protecting group, and a is
`1, 2, 3, or 4. FG is a fluorescent dye moiety which emits in the
`visible or near infrared spectrum. R is each independently H
`or C-C alkyl. V is —C(O)— or —NRC(O)— or NRC
`(S)-. W is NRC(O) , NRC(O)NR NRC(S)NR ,
`- NRC(O)O - OC(O)NR - OC(O) –C(O)NR-,
`or –C(O)O - Y is C(O) , NRC(O)-, - NRC(S) ,
`—OC(O). In exemplary embodiments m is 1, 2, 3, 4, 5, or 6:
`n is 1,2,3,4, 5 or 6; p is 0, 1, 2, or 3, and when p is 2 or 3, each
`R" may be the same or different. R' is H, C-C alkyl, C-C,
`aryl, or C-C alkylaryl. R and R are independently H,
`CO.H, or CO.R, where R is a C-C alkyl, C-C aryl, or
`C-C alkylaryl, wherein when one of RandR is CO, Hor
`COR, the other is H. In some embodiments, the fluorescent
`dye moiety emits in the near infrared spectrum.
`0029 Embodiments include compounds having the struc
`ture
`
`R
`A-W
`R
`NN19
`V -cis--
`
`N
`
`A
`A-W
`
`R1
`
`R
`N
`
`R3
`
`(CH2)
`
`R
`
`O
`
`* R2
`
`W-(CH)-Y-N
`Y(CH.), l Z
`QOC
`N
`N
`COQ
`
`wherein Z is tetrazole or COQ; each Q is independently
`selected from hydrogen or a protecting group, and a is 1, 2, 3,
`or 4. One of A and A' is Chand the other is FG, where FG is
`a fluorescent dye moiety which emits in the visible or near
`infrared spectrum and Ch is metal chelating moiety option
`ally including a chelated metal. R is each independently Hor
`C-C alkyl. V or V" are independently —C(O)— —NRC
`(O) , or - NRC(S)-. W is
`NRC(O)-, - NRC(O)
`NR NRC(S)NR
`NRC(O)O
`OC(O)NR
`OC(O) , —C(O)NR , or –C(O)O - Y is C(O) ,
`NRC(O) ,
`NRC(S) ,
`OC(O). In exemplary
`
`embodiments m is 1, 2, 3, 4, 5, or 6; n is 1, 2, 3, 4, 5 or 6; and
`g is 1,2,3,4, 5, or 6; p is 0, 1, 2, or 3, and when p is 2 or 3, each
`R" may be the same or different. R' is H. C-C alkyl, C-C,
`aryl, or C-C alkylaryl. R and R are independently H,
`CO.H, or CO.R, where R is a C-C alkyl, C-C aryl, or
`C-C alkylaryl, wherein when one of RandR is COH or
`CO.R, the other is H. In some embodiments, the fluorescent
`dye moiety emits in the near infrared spectrum. Some
`embodiments further include a chelated metal.
`0030 Embodiments include compounds having the struc
`ture
`
`R
`
`N
`
`R1
`
`O
`
`Gl
`
`R3
`
`(CH2)
`
`2
`pR
`
`R
`
`W Tz-i (CH2)-Y-N
`g
`
`(CH2)
`
`usul.
`
`QOC
`
`N
`H
`
`N
`H
`
`Z
`
`CO
`2O
`
`Petitioner GE Healthcare – Ex. 1005, p. 33
`
`

`

`US 2012/0009121 A1
`
`Jan. 12, 2012
`
`wherein subunits associated with p, q, r, and S may be in any
`order. Z is tetrazole or CO.Q; each Q is independently
`selected from hydrogen or a protecting group, and a is 1, 2, 3,
`or 4. R is each independently H or C-C alkyl. In this exem
`plary embodiment r is 1. TZ is a triazole group having the
`Structure
`
`L- 2 L2
`
`O
`
`NN Q-- --
`-Qu
`
`NN
`
`Ll
`
`S. N-L2
`
`where
`
`L is --all- O
`R5
`
`NRC(O)NR NRC(S)NR , or
`NRC(O) ,
`X is
`NRC(O)O : X is C(O)NR - NRC(O)NR , NRC
`(S)NR , or OC(O)NR : R is H, COH, or COR,
`where R is a C-C alkyl, Ca-Caryl, or C-C alkylaryl; b
`is 1,2,3, or 4, and dis 1,2,3, or 4. In exemplary embodiments
`q is 0 or 1, W is NRC(O)-, - NRC(O)NR NRC(S)
`NR-, - NRC(O)O
`—OC(O)NR
`—OC(O) ,
`—C(O)NR , or—C(O)O ; n is 1,2,3,4,5 or 6; and Rand
`R are independently H, COH, or COR, where R is a
`C-C alkyl, C-C aryl, or C-C alkylaryl, wherein if one
`of R and R is COH or COR, then the other is H. In
`exemplary embodiments s is 0 or 1; Y is C(O)— —NRC
`(O)— —NRC(S)— —OC(O); and m is 1, 2, 3, 4, 5, or 6. In
`exemplary embodiments p is 0, 1, 2, or 3, and when p is 2 or
`3, each R' may be the same or different; and R' is H. C-C,
`alkyl, C-C2aryl, or C-C alkylaryl. G' is a moiety selected
`from the group consisting of
`
`V.
`
`V.
`
`ch1 S.X. Fo1 S.X. and
`
`R
`
`R
`
`R
`
`A scontinued
`N NH
`Y CH
`N-(CH),
`
`A-W
`
`R
`
`l
`
`X s
`
`O
`
`where Ch is a metal chelating moiety, optionally including a
`chelated metal; FG is a fluorescent dye moiety which emits in
`the visible or near infrared spectrum; one of A and A' is Ch
`and the other is FG: V and V are each independently
`C(O) , – NRC(O) - NRC(S)-, or - OC(O)-; and
`g is 1, 2, 3, 4, 5, or 6. In some embodiments, the fluorescent
`dye moiety emits in the near infrared spectrum. Some
`embodiments include a chelated metal.
`0031. Embodiments of the invention include methods of
`imaging one or more cells, organs or tissues by exposing the
`cell to or administering to a organism an effective amount of
`a compound discussed above, where the compound includes
`a fluorescent dye moiety, or a metal isotope Suitable for imag
`1ng.
`0032 Embodiments of the invention include methods of
`treating a tumor comprising administering a therapeutically
`effective amount of a compound discussed above, where the
`compound includes a therapeutically effective radioisotope.
`0033 Embodiments of the invention include methods for
`sorting cells by exposing the cells to a compound discussed
`above, where the compound includes a fluorescent dye moi
`ety, followed by separating cells which bind the compound
`from cells which do not bind the compound.
`0034 Embodiments of the invention include methods of
`intraoperative tumor mapping comprising administering an
`effective amount of a compound discussed above to a Subject,
`where the compound includes a fluorescent dye moiety.
`
`BRIEF DESCRIPTION OF THE DRAWINGS
`0035 FIG. 1 shows SPECT-CT images of a PSMA+
`LNCaP tumor-bearing mouse injected intravenously with
`exemplary compound "TclSRV32.
`0036 FIG. 2. GE explore VISTA pseudodynamic PET
`image (co-registered with the corresponding CT image) of a
`PSMA+ LNCaP tumor-bearing mouse injected intravenously
`with 0.2 mCi (7.4 MBq) of exemplary compound Ga
`SRV27.
`0037 FIG. 3. GE explore VISTA PET image (co-regis
`tered with the corresponding CT image) of a PSMA+ PIP and
`PSMA-flu tumor-bearing mouse injected intravenously with
`0.2 mCi (7.4 MBq) of exemplary compound GaSRV 100.
`0038 FIG. 4 shows a synthetic scheme for exemplary
`compound SRV 100 and '''InSRV 100.
`0039 FIG. 5 shows SPECT-CT images of a PSMA+ PC-3
`PIP tumor-bearing mouse injected intravenously with exem
`plary compound '''InSRV27.
`0040 FIG. 6 shows SPECT-CT images of a PSMA+ PC-3
`PIP tumor-bearing mouse injected intravenously with exem
`plary compound '''InSRV 100.
`0041 FIG. 7 shows SPECT-CT images of a PSMA+ PC-3
`PIP tumor-bearing mouse injected intravenously with exem
`plary dual modality compound '''InSRV73.
`0042 FIG. 8 shows the absorbance and emission spectra,
`and quantum yield of exemplary compound YC-27.
`
`Petitioner GE Healthcare – Ex. 1005, p. 34
`
`

`

`US 2012/0009121 A1
`
`Jan. 12, 2012
`
`0043 FIG. 9 shows the fluorescence decay of exemplary
`compound YC-27.
`0044 FIG. 10 shows an ICso curve of compound YC-27
`using a fluorescence-based NAALADase assay
`004.5
`FIG. 11 shows in vivo imaging of a NOD/SCID
`mouse (mouse #1), bearing PC3-PIP (forward left flank) and
`PC3-flu (forward right flank) tumors. Mouse #1 received 10
`nmol of YC-27 and dorsal (animal prone) and ventral (animal
`supine) views were obtained. Dorsal and ventral views at 40
`min p.i. (A, B, respectively); 18.5 h (C, D); 23 h (E. F); 42.5
`h (G,H); 68 h (I,J). Dorsal view of pre-injection image (K).
`Dorsal and ventral views 70.5 h p. i. (L. M). Images after
`midline laparotomy (N) and individually harvested organs
`(O) on a Petri dish at 70.5 h p.i. Images were scaled to the
`same maximum (arbitrary units).
`0046 FIG. 12 shows in vivo imaging of a NOD/SCID
`mouse (mouse #2) (left panel), bearing PC3-PIP (forward left
`flank) and PC3-flu (forward right flank) tumors. Mouse #2
`received 1 nmol of YC-27 and dorsal (animal prone) and
`ventral (animal supine) views were obtained. Dorsal and ven
`tral views of the pre-injection image (A, B, respectively): 10
`min p.i. (C, D); 20.5 h (E. F); 24 h (G, H). Images after
`midline laparotomy (I) and individually harvested organs (J)
`on a Petri dish at 24 hp.i. Right Panels: Mouse #3 in same
`orientation as mouse #2. Mouse #3 received 1 nmol of YC-27
`co-injected with 1 umol of DCIBZL, which served as a block
`ing agent to test binding specificity. Images were scaled to the
`same maximum (arbitrary units).
`0047 FIG. 13 shows SPECTCT images of a PSMA+
`LNCaP tumor-bearing mouse injected intravenously with
`exemplary compound "TcISRVI34B.
`0048 FIG. 14 shows SPECTCT images of a PSMA+
`PC3-PIP tumor-bearing mouse injected intravenously with
`exemplary compound "TcISRVI34B.
`0049 FIG. 15 shows SPECTCT images of a PSMA+
`PC3-PIP (forward left flank) and PSMA+ PC3-flu (forward
`right flank) tumor-bearing mouse injected intravenously with
`exemplary compound "TclSRVI34A.
`0050 FIG. 16 shows SPECTCT images of a PSMA+
`PC3-PIP (forward left flank) and PSMA- PC3-flu (forward
`right flank) tumor-bearing mouse injected intravenously with
`exemplary compound "TcISRVI34B.
`0051 FIG. 17 shows PC3-PIP and PC3-flu cells treated
`with fluorescent compound YC-VIII-36 (green, top left) and
`DAPI (blue), and PC3-PIP and PC3-flu cells treated with both
`YC-VIII-36 and PSMA inhibitor, PMPA.
`0.052
`FIG. 18 shows PC3-PIP cells treated with DAPI
`(blue) and varying concentrations of YC-VIII-36 (green).
`0053 FIG. 19 shows time dependent internalization of
`YC-VIII-36 into PC3-PIP cells treated with YC-VIII-36
`(green) and DAPI (blue).
`0054 FIG. 20 shows titration and detection of varying
`amounts of YC-VIII-36 injected subcutaneously into a nude
`mouse. (IVIS spectrum with 10 second exposure followed by
`spectral unmixing)
`0055 FIG. 21 shows fluorescence images of a PSMA+
`PC3-PIP and PSMA-PC3-flu tumor-bearing mouse injected
`intravenously with exemplary compound YC-VIII-36.
`0056 FIG. 22 shows fluorescence images of a PSMA+
`PC3-PIP and PSMA-PC3-flu tumor-bearing mouse injected
`intravenously with exemplary compound YC-VIII-36 180
`minutes after injection (top) and biodistribution of exemplary
`compound YC-VIII-36 180 minutes after injection (botto