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`Application Number: 1 8354282
`
`Document Date: 07/1 8/2023
`
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`Form Revision Date: March 1, 2019
`
`Petitioner GE Healthcare – Ex. 1002, p. 1
`
`

`

`Doc Code: TRACK1.REQ
`Document Description: TrackOne Request
`
`PTO/A IA/424 (04-14)
`
`CERTIFICATION AND REQUEST FOR PRIORITIZED EXAMINATION
`UNDER 37 CFR 1.102(e) (Page 1 of 1)
`
`First Named
`Inventor:
`Title of
`Invention:
`
`Xing YANG
`
`Nonprovisional Application Number (if
`known):
`
`IMAGING AND RADIOTHERAPEUTICS AGENTS TARGETING FIBROBLAST-ACTIVATION PROTEIN-ALPHA (FAP-ALPHA)
`
`APPLICANT HEREBY CERTIFIES THE FOLLOWING AND REQUESTS PRIORITIZED EXAMINATION FOR
`THE ABOVE-IDENTIFIED APPLICATION.
`
`1. The processing fee set forth in 37 CFR 1.17(i)(1) and the prioritized examination fee set forth in
`37 CFR 1.17(c) have been filed with the request. The publication fee requirement is met
`because that fee, set forth in 37 CFR 1.18(d), is currently $0. The basic filing fee, search fee,
`and examination fee are filed with the request or have been already been paid. I understand
`that any required excess claims fees or application size fee must be paid for the application.
`
`2. I understand that the application may not contain, or be amended to contain, more than four
`independent claims, more than thirty total claims, or any multiple dependent claims, and that
`any request for an extension of time will cause an outstanding Track I request to be dismissed.
`
`3. The applicable box is checked below:
`
`I.
`
`~ Ori i nal A
`
`l i c a t ion Track One - Prioritized Examination under 1 . 102 e 1
`
`(a) The application is an original nonprovisional utility application filed under 35 U.S.C. 111(a).
`This certification and request is being filed with the utility application via EFS-Web.
`— -OR —(cid:173)
`(b) The application is an original nonprovisional plant application filed under 35 U.S.C. 111(a).
`This certification and request is being filed with the plant application in paper.
`
`An executed inventor's oath or declaration under 37 CFR 1.63 or 37 CFR 1.64 for each
`inventor, or the application data sheet meeting the conditions specified in 37 CFR 1.53(f)(3)(i) is
`filed with the application.
`
`II.
`
`Re uest f or Continued Examination - Prioritized Examination under 1 . 102 e 2
`
`A request for continued examination has been filed with, or prior to, this form.
`If the application is a utility application, this certification and request is being filed via EFS-Web.
`The application is an original nonprovisional utility application filed under 35 U.S.C. 111(a), or is
`a national stage entry under 35 U.S.C. 371.
`iv. This certification and request is being filed prior to the mailing of a first Office action responsive
`to the request for continued examination.
`v. No prior request for continued examination has been granted prioritized examination status
`under 37 CFR 1.102(e)(2).
`
`„„„„„/Jeffrey W. Childers/
`;;;„;, „Jeffrey W. Childers
`
`„„2023-07-18
`oII Number58 1 26
`
`Note: Th is form must be signed in accordance with 37 CFR 1.33. See 37 CFR 1.4(d) for signature requirements and certificat/'ons.
`Submit multi le formsifmore than one si natureis re uired. *
`*Total of 1
`
`forms are submitted.
`
`Petitioner GE Healthcare – Ex. 1002, p. 2
`
`

`

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`
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`
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`9. A record from this system of records may be disclosed, as a routine use, to a Federal, State, or local law
`enforcement agency, if the USPTO becomes aware of a violation or potential violation of law or regulation.
`
`Petitioner GE Healthcare – Ex. 1002, p. 3
`
`

`

`Attorney Docket: JHU-36631.303
`
`IMAGING AN D R A D I O T H E R A P EU T ICS AG E NTS TA R G E T IN G F I B RO B L A ST (cid:173)
`
`A CTIVA T IO N PR O T E I N - A L PHA ( FA P-AL PH A )
`
`C R O S S -RE F E R E N C E TO R E L A T E D A P P L IC A T IO N S
`
`This application is a continuation of U.S. Patent Application 16/7S8,182, filed
`
`April 22, 2020, which is a U.S. )371 National Entry Application of
`
`PCT/US2018/OS7086, filed October 23, 2018, which claims the benefit of U.S.
`
`Provisional Application No. 62/S7S,607, filed October 23, 2017, each of which is
`
`incorporated herein by reference in its entirety.
`
`FEDE R A L L Y S P O N S O R E D R E S E A R C H O R D E V E L O P M E N T
`
`This invention was made with <government support under CA197470 awarded
`
`b y the National Institutes of Health. T h e < overnment has certain ri< hts in the
`
`invention.
`
`B ACKGROU N D
`
`Fibroblast-activation protein-u (FAP-u) expression has been detected on the
`
`surface of fibroblasts in the stroma surroundin < >90% of the epithelial cancers
`
`examined, includin < mali< nant breast, colorectal, skin, prostate and pancreatic
`
`cancers. (Garin-Chesa, et al., 1990; Retti <, et al., 1993; Tuxhorn, et al., 2002;
`
`Scanlan, et al., 1994). It is a characteristic marker for carcinoma-associated-fibroblast
`
`(CAF), which plays a critical role in promotin <~ an<~io<~enesis, proliferation, invasion,
`
`and inhibition of tumor cell death. (A l l i nen, et al., 2004; Franco, et al., 2010). In
`
`healthy adult tissues, FAP-u expression is only limited to areas of tissue remodelin <
`
`or wound healin <~. (Scanlan, et al., 1994; Yu, et al., 2010; Bae, et al., 2008; Kraman,
`
`et al., 2010). In addition, FAP-u-positive cells are observed durin < embryo< enesis in
`
`areas of chronic inflammation, arthritis, and fibrosis, as well as in soft tissue and bone
`
`sarcomas. (Scanlan, et al., 1994; Yu, et al., 2010). These characteristics make FAP-u
`
`a potential ima < in< and radiotherapeutic tar< et for cancer and inflammation diseases.
`
`Because FAP-u is expressed in tumor stroma, anti-FAP antibodies have been
`
`investi<~ated for radioimmunotar <~etin<~ of mali<~nancies, includin<~ murine F19,
`
`sibrotuzumab (a humanized version of the F19 antibody), ESC11, ESC14, and others.
`
`(Welt, et al., 1994; Scott, et al., 2003; Fischer, et al., 2012). A n t i bodies also
`
`demonstrated the feasibility of ima <~in<~ inflammation, such as rheumatoid arthritis.
`
`Petitioner GE Healthcare – Ex. 1002, p. 4
`
`

`

`Attorney Docket: JHU-36631.303
`
`(Laverman, et al., 2015). The use of antibodies as molecular ima < in< a< ents,
`
`however, suffers from pharmacokinetic limitations, including slow blood and non(cid:173)
`
`tar< et tissue clearance (normally 2 — 5 days or ion < er) and non-specific or < an uptake.
`
`Low molecular wei < ht (LMW) a < ents demonstrate faster pharmacokinetics and a
`
`higher specific signal within clinically convenient times after administration. T h ey
`
`also can be synthesized in radiolabeled form more easily and may offer a shorter path
`
`to re< ulatory approval. (C oenen, et al., 2010; Coenen, et al., 2012; Reilly, et al.,
`
`2015). To date, however, no LM W
`
`l i g and has been reported with ideal properties for
`
`nuclear imaging of FAP-u.
`
`In some aspects, the presently disclosed subject matter provides a compound
`
`S UMM A R Y
`
`of Formula (I):
`
`B
`
`L
`
`A
`
`wherein: A i s a targeting moiety for FAP-u; B is any optical or radiolabeled
`
`functional group suitable for optical imagin <~, PET imagin<~, SPECT imagin <~, or
`
`radiotherapy; and L is a linker having bi-functionalization adapted to form a chemical
`
`bond with B and A.
`
`In particular aspects, A is an FAP-u tar < etin< moiety havin< the structure of:
`
`y (R > x)
`
`( 2x) y
`
`y ( » ' )
`
`N
`
`(R3x)y
`
`C H j
`V
`
`4x
`
`R5x
`
`R6x
`
`R7x
`
`(X');
`
`wherein each y is independently an integer selected from the group consisting
`
`of 0, 1, and 2; Ri „ R 2 „ a n d R ; „ a r e each independently selected from the< roup
`
`-O-Ci<alkyl, and -S-Ci(cid:173) (alkyl; Rix is selected
`consisting of H, OH, halo < en, Ci-(alk y l ,
`-CN, -B(OH)2, -C(O)alkyl, -C(O)aryl-, -C = C(cid:173)
`from the group consisting of H,
`C(O)aryl, -C = C-S(O)2aryl, -CO2H,
`H; R~„R<,„ and R7, are each independently selected from the < roup consistin< of H,
`
`-SO;H, -SO2NH2, -PO;H2, and 5-tetrazolyl; Rz x is
`
`OH, oxo, halogen, -Ci <alkyl, -0-Ci <alkyl,
`
`-S-Ci <alkyl, -NRxxR>x, -OR i2x, -Het2 and(cid:173)
`
`Ar2; each of Ci <,alkyl being optionally substituted with from 1 to 3 substituents
`
`Petitioner GE Healthcare – Ex. 1002, p. 5
`
`(cid:173)
`

`

`Attorney Docket: JHU-36631.303
`
`selected from -OH and halo < en; Rx„R > „ a n d Ri 2x are each independently selected
`
`from the <~roup consistin <~ of H, -OH, halo, -Ci <alkyl, -0-Ci <alkyl, -S-Ci <alkyl, and(cid:173)
`
`Ar;; Rni„ R i
`
`i „ Ri ; x and Ri<x are each independently selected from the < roup
`
`consistin<~ of H, -OH, halo <~en, -Ci <alkyl, -0-Ci <alkyl, and -S-Ci <alkyl; Ari, Ar2 and
`
`Ar; are each independently a 5- or 6-membered aromatic monocycle optionally
`
`comprisin <~ 1 or 2 heteroatoms selected from 0, N and S; each of Ari, Ar2 and Ar;
`
`bein<~ optionally and independently substituted with from 1 to 3 substituents selected
`
`from - N R n i xRi ix, -Ci <alkyl, -0-Ci <alkyl, and -S-Ci <alkyl; Het2 is a 5- or 6(cid:173)
`
`membered non-aromatic monocycle optionally comprisin <~ 1 or 2 heteroatoms
`
`selected from 0, N and S; Het2 bein <~ optionally substituted with from 1 to 3
`
`substituents selected from -NR i ; xRiz„ - C i <alkyl,
`
`- 0 - C / -( al k y l , an d - S - C f (cid:173)
`
`(alkyl; v is
`
`0, 1,2, or3; and
`
`represents a 5 to 10-membered N-containin <~ aromatic or non-aromatic mono- or
`
`bicyclic heterocycle, said heterocycle optionally further comprisin <~ 1, 2 or 3
`
`heteroatoms selected from 0, N and S; wherein ~
`
`indi c a t es a point of attachment
`
`of the FAP-u bindin <~ li<~and to the linker, L, or the reporter moiety, B, wherein the
`
`point of attachment can be throu <~h any of the carbon atoms of the 5 to 10-membered
`
`N-containin<~ aromatic or non-aromatic mono- or bicyclic heterocycle thereof; and
`
`stereoisomers and pharmaceutically acceptable salts thereof.
`
`In more particular aspects, A is an FAP-u tar < etin< mo i ety h a v i n < t h e
`
`structure of:
`
`y(R>x)
`
`( 2x)y
`
`y(R3x')
`
`N
`
`C
`
`6&
`7 +
`
`8
`
`N
`
`wherein ~
`
`indi c a t es a point of attachment of the FAP-u bindin< li< and to th e
`
`linker, L, or the reporter moiety, B, wherein the point of attachment can be throu <~h
`
`any of carbon atoms 5, 6, 7, or 8 of the quinolinyl rin <~ thereof; and stereoisomers and
`
`pharmaceutically acceptable salts thereof.
`
`In yet more particular aspects, A is selected from the <~roup consistin<~ of:
`
`Petitioner GE Healthcare – Ex. 1002, p. 6
`
`

`

`Attorney Docket: JHU-36631.303
`
`0
`
`N ~
`
`0
`
`0
`
`N~
`
`0
`
`0
`
`N~
`
`0
`
`A~
`
`N
`
`N
`
`A2
`
`;an d
`
`A3
`
`N
`
`In other aspects, the presently disclosed subject matter provides a
`
`pharmaceutical composition comprisin <~ a compound of formula (I).
`
`In some aspects, the presently disclosed subject matter provides a method for
`
`ima< in< a disease or disorder associated with fibroblast-activation protein-u (FAP-u),
`
`the method comprisin <~ administerin<~ a compound of formula (I), wherein the
`
`compound of formula (I) comprises an optical or radiolabeled functional <~roup
`
`suitable for optical ima <~in<~, PET ima<~in<~, or SPECT
`
`i m a<~in<~; and obtain in<~ an
`
`ima< e.
`
`In other aspects, the presently disclosed subject matter provides a method for
`
`inhibitin <~ fibroblast-activation protein-u (FAP-u), the method comprisin<~
`
`administerin < to a subject in need thereof an effective amount of a compound of
`
`formula (I).
`
`In yet other aspects, the presently disclosed subject matter provides a method
`
`for treatin< a fibroblast-activation protein-u (FAP-u)-related disease or disorder, the
`
`method comprisin < administerin< to a subject in need of treatment thereof an effective
`
`amount of a compound of formula (I), wherein the compound of formula (I)
`
`comprises a radiolabeled functional < roup suitable for radiotherapy.
`
`In certain aspects, the (FAP-u)-related disease or disorder is selected from the
`
`<~roup consistin<~ of a proliferative disease, includin<~, but not limited to, breast cancer,
`
`colorectal cancer, ovarian cancer, prostate cancer, pancreatic cancer, kidney cancer,
`
`lun<~ cancer, melanoma, fibrosarcoma, bone and connective tissue sarcomas, renal cell
`
`carcinoma, <pliant cell carcinoma, squamous cell carcinoma, and adenocarcinoma;
`
`diseases characterized by tissue remodelin <~ and/or chronic inflammation; disorders
`
`involvin<~ endocrinolo <~ical dysfunction; and blood clottin<~ disorders.
`
`Certain aspects of the presently disclosed subject matter havin < been stated
`
`hereinabove, which are addressed in whole or in part by the presently disclosed
`
`subject matter, other aspects will become evident as the description proceeds when
`
`taken in connection with the accompanyin < Examples and Fi < ures as best described
`
`herein below.
`
`Petitioner GE Healthcare – Ex. 1002, p. 7
`
`

`

`Attorney Docket: JHU-36631.303
`
`B RIEF DESCRIPTION OF TH E F I G U R E S
`
`The patent or application file contains at least one drawin < executed in color.
`
`Copies of this patent or patent application publication with color drawin <~s will be
`
`provided by the Office upon request and payment of the necessary fee.
`
`Havin< thus described the presently disclosed subject matter in < eneral terms,
`
`reference will now be made to the accompanyin <~ Fi<~ures, which are not necessarily
`
`drawn to scale, and wherein:
`
`FIG. 1A, FIG. 1B, and FIG. 1C show the synthetic pathway and structures of
`
`representative FAP-tar < eted a< ents, XY-FAP -01 and ]"' I n ] - X Y - F A P - 0 2. F I G . 1 A
`
`shows the multi- step synthesi s of the 1 i<~and precursor, tert-butyl(S)-(3-((4-((2-(2(cid:173)
`cyanopyrrol i di n-1-yl )-2(cid:173) oxoethyl ) carb am oyl )qui nol i n-6-yl ) oxy)propyl )carbamate.
`After each step, the reaction mixture was loaded onto a 25(cid:173) < C18 cartrid< e and
`
`purified with a MeCN/water/TFA < radient. Identity of intermediate products was
`
`confirmed with 'H N M R . F I G . 1B shows the full structure of optical ima<~in<~ a<~ent,
`
`XY-FAP-01. X Y - F A P -01 was produced with a one step reaction between the
`
`precursor and IRDye800CW- N HS. The major product was obtained at a yield of 85%
`
`after purification with HPLC . F I G . 1C shows the full structure of the SPECT ima<~in<~
`
`a<~ent, ]" ' I n ] - X Y - F A P - 02 . F i r st, the precursor was functionalized with DOTA vi a a
`
`one step reaction between the precursor and DOTA - G A ( t - B u)q-NHS. U n l abeled
`
`product was purified via HPLC to produce XY- F A P-02. Subsequent radiolabelin <
`
`w ith " ' I n and HPLC purification resulted in the radiolabeled product, ]" ' I n ] - X Y (cid:173)
`
`FAP-02;
`
`FIG. 2 shows the inhibitory activity of XY - F A P - 01 on human recombinant
`
`FAP. The inhibitory activity of XY - F A P - 01 was determined usin< ~ a flu o r o<geni FA P
`
`assay kit. Enzymatic activity of human recombinant FAP on a native substrate was
`
`inhibited in a concentration dependent fashion by XY - F A P - 0 1. Se mi-lo<~ inhibitory
`
`curves of XY- F A P - 01 activity were < enerated and the determined Ki value of X Y (cid:173)
`FAP-01 was 1.26 nM;
`
`FIG. 3A, FIG. 3B, and FIG. 3C show the assessment of thei»
`
`ii / n ~ bindin <~
`
`ability and specificity of XY - F A P - 01 and ]" ' I n ] - X Y - F A P - 0 2. F I G . 3A shows the
`
`concentration dependent uptake of XY - F A P - 01 in various cell lines. Cells incubated
`
`with various concentrations (ran<ate: SO nM to 0.78 nM) of XY - F A P - 01 were ima<~ed
`
`with the LI-COR Pearl Impulse Ima < er to assess uptake of a < ent in various FAP(cid:173)
`
`positive and FAP-ne < ative cell lines (left). D o se-response curves of X Y- F A P - 0 1
`
`Petitioner GE Healthcare – Ex. 1002, p. 8
`
`

`

`Attorney Docket: JHU-36631.303
`
`uptake in FAP-positive cell lines (NCIH2228, U87, and SKM E L 24) and FAP(cid:173)
`
`ne<~ative cell lines (PC3, NCIH226, and HCT 116) were <~enerated (ri<~ht). FIG. 3B
`
`shows the inhibition of XY - F A P - 01 uptake in FAP-positive cell-lines. Cells
`
`incubated with 25-nM XY - F A P - 01 were incubated with various concentrations of
`
`either a DPPIV and FAP inhibitor, Talabostat, or a DPPIV-only inhibitor, Sita <~liptin.
`
`Uptake of XY - F A P - 01 was measured and semi-lo< inhibitor-response curves were
`
`< enerated for both Talabostat and Sita < liptin. FIG. 3C shows the uptake of ] " ' I n ] (cid:173)
`
`XY-FAP-02 in FAP-positive U87 and FAP-ne<~ative PC3 cell lines. Cells were
`
`incubated with 1 pCi ] ' "
`
`I n ] - X Y - F A P - 02 and were washed with cold PBS.
`
`Radioactivity of the cell pellets was measured and normalized to the incubated dose;
`
`FIG. 4 is a table showin < the ex ii n~ tissue biodistribution of ]'"
`
`I n ] - X Y - F A P (cid:173)
`
`01 in tumor bearin <~ mice. At 5 min, 0.5 h, 2 h, 6 h, and 12 h after injection of 10 pCi
`
`]"'In]-X Y - F A P - 01, NOD /S KID m i ce bearin<~ U87 and PC3 tumor xeno<~rafts were
`
`sacrificed and tissues were collected for biodistribution analysis. Additionally, mice
`
`co-injected with unlabeled XY-F A P -02 and 10 pCi ] " ' I n ] - X Y - F A P - 0 1 were
`
`sacrificed at 6 h post-injection to study the effect of blockin < on uptake of the
`
`radiolabeled compound. Data presented as mean + standard deviation. "Student's t test
`
`comparison of mean %ID/ <~ of PC3 tumor versus U87 tumor demonstrated si<~nificant
`
`difference between the two < roups at 5 min, 0.5 h, 2 h, and 6 h post injection
`
`(p<0.0001). No si < nificant difference between the two < roups were seen in the
`
`blockin <~ study at 6 h. S t u dent's t test comparison of mean %ID/<~ of PC3 tumor
`
`versus U87 tumor demonstrated si < nificant difference between the two < roups at 12 h
`
`post injection (p = 0.0006).
`
`'Student's t test comparin<~ %ID/<~ between PC3 tumor and
`
`U87 tumors at 6 h post injection showed si <~nificant difference between %ID/<~ tumors
`
`in the blockin <~ study at 6 h versus the normal biodistribution results at 6 h
`
`(p<0. 0001);
`
`FIG. 5A and FIG. 5B show the time-activity relationship of the ex ii n~
`
`biodistribution of ] " ' I n ] - X Y - F A P - 02. FIG. 5A shows tissue time activity curves
`
`(TACs) of ] " ' I n ] - X Y - F A P -02 activity in U87 tumor, PC3 tumor, and blood. FIG. 5B
`
`s hows the ratios of % I D / <' between U87 tumor and PC3 tumor, blood, and muscle
`
`(mm) versus time;
`
`FIG. 6 shows serial NIRF-ima <~in<~ of XY-FAP-01 in tumor bearin<~ mice.
`
`NOD/SKID m ice bearin <~ FAP-positive U87 (yellow circle) and FAP-ne<~ative PC3
`
`(red circle) tumor xeno <~rafts were injected with 10 nmol of X Y - F A P -01 via the tail
`
`Petitioner GE Healthcare – Ex. 1002, p. 9
`
`

`

`Attorney Docket: JHU-36631.303
`
`vein followed by serial NIRF-ima<~in<~ on the LI-COR Pearl Impulse Ima<~er.
`Representative ima <~es at 0.5 h, 1 h, 2.5 h, and 4 h after injection are shown;
`
`FIG. 7 shows SPECT-CT i m a<~es of ]"' I n ] - X Y - F A P -02 at 30 min, 2 h, 6 h,
`
`and 24 h after injection in NOD/S KID fe male mice bearin< U87 and PC3 tumor
`
`xeno<~rafts in the upper flanks; and
`
`FIG. 8 show three-dimensional SPECT-CT i m a<~es of ]"' I n ] - X Y - F A P -02 at
`
`30 min, 2 h, 6 h, and 24 h after injection in NOD /S KID fe male mice bearin<~ U87 and
`
`PC3 tumor xeno<~rafts in the upper flanks.
`
`DETAILED D E SCRIPTION
`
`The presently disclosed subject matter now will be described more fully
`hereinafter with reference to the accompanyin <~ Fi<~ures, in which some, but not all
`
`embodiments of the presently disclosed subject matter are shown. L i k e numbers refer
`
`to like elements throu < hout. The presently disclosed subject matter may be embodied
`
`in many different forms and should not be construed as limited to the embodiments
`
`set forth herein; rather, these embodiments are provided so that this disclosure will
`
`satisfy applicable le < al requirements. Indeed, many modifications and other
`
`embodiments of the presently disclosed subject matter set forth herein will come to
`
`mind to one skilled in the art to which the presently disclosed subject matter pertains
`
`havin< the benefit of the teachin < s presented in the fore < oin< descriptions and the
`
`associated Fi < ures. Therefore, it is to be understood that the presently disclosed
`
`subject matter is not to be limited to the specific embodiments disclosed and that
`
`modifications and other embodiments are intended to be included within the scope of
`
`the appended claims.
`
`I MAGING AN D R A D I O T H E R A P E U T ICS AG E NTS TA R G E T I N G
`
`FIBROBLAST-ACT I V A T I O N P R O T E I N-0 (FAP-0)
`
`FAP-u is a type II inte < ral membrane serine protease of the prolyl
`
`oli< opeptidase family, which are distin < uished by their ability to cleave the Pro-AA
`
`peptide bond (where AA represents any amino acid). It has been shown to play a role
`
`in cancer by modifyin <~ bioactive si<~nalin<~ peptides throu <~h this enzymatic activity
`
`(Kelly, et al., 2005; Edosada, et al., 2006). FA P -u expression has been detected on
`
`the surface of fibroblasts in the stroma surroundin «r e a ter than 90% of the epithelial
`
`cancers, includin <~, but not limited to, mali<~nant breast, colorectal, skin, prostate,
`
`Petitioner GE Healthcare – Ex. 1002, p. 10
`
`

`

`Attorney Docket: JHU-36631.303
`
`pancreatic cancers, and the like, and inflammation diseases, includin <, but not limited
`
`to, arthritis, fibrosis, and the like, with nearly no expression in healthy tissues.
`
`Accordin <~ly, ima<~in<~ and radiotherapeutic a <~ents specifically tar<~etin<~ FAP-u is of
`
`clinical importance.
`
`FAP-u exists as a homodimer to carry out its enzymatic function. Inhibitors
`
`selectively tar < etin< FAP-u has been reported (Lo, et al., 2009; Tsai, et al., 2010;
`
`Ryabtsova, et al., 2012; Poplawski, et al., 2013; Jansen, et al., 2013; Jansen, et al.,
`
`2014). The presently disclosed subject matter provides, in part, a FAP-u selective
`
`tar< etin< moiety that can be modified with an optical dye, a radiometal chelation
`
`complex, and other radiolabeled prosthetic <groups, thus providin <~ a platform for the
`ima<~in<~ and radiotherapy tar<~etin<~ FAP-u.
`
`Radionuclide molecular ima <~in<~, includin<~ positron emission tomo<~raphy
`(PET), is the most mature molecular ima < in< technique without tissue penetration
`limitations. Due to its advanta < es of hi< h sensitivity and quantifiability, radionuclide
`
`molecular ima <~in<~ plays an important role in clinical and preclinical research (Youn,
`
`et al., 2012; Chen, et al., 2014). M any radionuclides, primarily P- and alpha emitters,
`
`have been investi < ated for tar< eted radioimmunotherapy and include both
`
`radiohalo < ens and radiometals (see Table 1 for representative therapeutic
`
`radi onucl i des).
`
`Table 1. Representative Therapeutic Radionuclides
`'"' R e '" " Re "C u
`'"Y
`' '
`I '" L u " ' Sm
`
`P-particle emitters
`
`'!
`
`'!
`
`'!
`
`'!
`
`'!
`
`'!
`
`-" - 'Pb
`
`'!
`
`u-particle emitters
`
`-'-"Ac, -"'Bi, -"-'Bi, -"' At, -"-'Pb
`
`Au<~er electron emitters
`
`l2 I
`
`'!
`
`l 2 I E7C l ill
`
`'!
`
`'!
`
`The hi <~hly potent and specific bindin<~ moiety tar<~etin<~ FAP-n enables its
`
`use in nuclear ima < in< and radiotherapy. The presently disclosed subject matter
`
`provides the first synthesis of nuclear ima <~in<~ and radiotherapy a<~ents based on this
`
`dual-tar< etin< moiety to FAP-n.
`
`Accordin <~ly, in some embodiments, the presently disclosed subject matter
`
`provides potent and selective low-molecular-wei <~ht (LMW)
`FAP-u selective inhibitor, conju < ated with a tar< etin< moiety feasible for
`
`l i <~ands of FAP-u, i.e., an
`
`modification with optical dyes and radiolabelin <~ <groups, includin <~ metal chelators
`
`Petitioner GE Healthcare – Ex. 1002, p. 11
`
`

`

`Attorney Docket: JHU-36631.303
`
`i o optical ima<~in<~, nuclear ima<~in<~(optical,
`and metal complexes, which enable ii»i
`PET and SPECT), and radiotherapy tar<~etin<~ FAP-u.
`Imp ortantly, the presently
`
`disclosed compounds can be modified, e. <~., conju<~ated with, labelin <~ <groups without
`
`si<~nificantly losin<~ their potency. The presently disclosed approach allows for the
`
`convenient labelin <~ of the FAP-u li<~and with optical dyes and PET or SPECT
`
`isotopes, includin < ~, but not limited to, '"Ga, ' Cu, '"F, "'Y , ' "Y , " ' Z r , " ' In, " " ' T c, '-" I,
`
`'-"I, for FAP-u related ima< in< applications. Further, the presently disclosed
`
`approach allows for the radiolabelin <~ of the FAP-u li<~and with radiotherapeutic
`I , - "'At, ' " I n , " ' S m , ' " ' R e ,
`' -" I , ' '
`isotopes, includin <~ but not limited to, '"Y, ' " L u ,
`-"-'Pb, -'-"Ac, -" 'Bi, -"-'Bi, -"-'Pb, and " Ga, for FAP-u related radio-therapy.
`'""Re, "Cu,
`
`In a particular embodiment, an optical a < ent conju< ated with IRDye-800CW
`
`(XY-FA P-01) was synthesized and showed selective uptakeii » i / i ' o on a FAP-u+
`
`U87 cell line and is»i i o on a FAP-u+ U 87 tumor and clearly detected the tumor, I n
`
`another particular embodiment, an ' " In labeled li <~and (XY-FAP-02-[" 'In)) was
`successfully obtained in hi <~h yield and purity from its precursor with a metal chelator.
`
`i o study showed clear tumor radiotracer uptake in mice bearin<~ FAP-u(cid:173)
`Their» i
`positive U87 tumors with minimum non-specific or <~an uptake, which allows the
`specific ima < in< of FAP-u expressin< tumors. The presently disclosed FAP-u
`
`tar< etin< moiety can be adapted for use with optical dyes and radioisotopes known in
`
`the art for ima< in< and therapeutic applications tar < etin< F A P - u .
`
`More particularly, in some embodiments, the presently disclosed subject
`
`matter provides a compound of the < eneral structure of Formula (I):
`B
`L
`A
`
`wherein: A i s a tar <~etin<~ moiety for FAP-u; B is any optical or radiolabeled
`
`functional <~roup suitable for optical ima<~in<~, positron-emiss'.:on tomo< raphy (PET)
`
`ima<~in<~, sin„~le-Iil:oton e:n i ssion co rn p u«;:d toino~ ia'<iliy (SPECT) ima <~in<~, or
`
`radiotherapy; and L is a linker havin <~ bi-functionalization adapted to form a chemical
`
`bond with B and A.
`
`Representative tar < etin< moieties for FAP-u are disclosed in U.S. Patent
`
`Application Publication No. US2014/03S76SO for Novel FAP Inhibitors to Jansen et
`
`al., published Dec. 4, 2014; U.S. Patent No. 9,346,814 for Novel FAP Inhibitors to
`
`Jansen et al., issued May 24, 2016; and International PCT Patent Publication No. WO
`
`2013/107820 for Novel FAP Inhibitors to Jansen et al., published July 2S, 2013, each
`
`of which are incorporate by reference in their entirety.
`
`Petitioner GE Healthcare – Ex. 1002, p. 12
`
`

`

`Attorney Docket: JHU-36631.303
`
`More particularly, U.S. Patent No. 9,346,814 to Jansen et al., discloses FAP-u
`
`inhibitors of formula (X), or a stereoisomer, tautomer, racemate, salt, hydrate, or
`
`solvate thereof, which are suitable for use with the presently disclosed subject matter:
`
`R ~x~~ ~ R 2x
`( , ~ R,„
`
`C H j
`V
`
`4x
`
`R5x
`
`R7x
`
`R6x
`
`(X);
`
`wherein:
`
`Rr x and R2x are each independently selected from the < roup consistin< of H,
`
`OH, halo <~en, Ci <alkyl, -0-Ci <alkyl, and -S-Ci<alkyl;
`
`R;, is selected from the <~roup consistin<~ of H, -CN, -B(OH)2, -C(0)alkyl, (cid:173)
`-PO-H2, and
`C(0)aryl-, -C = C-C(0)aryl, -C = C-S(0)2aryl, - C0 2H , -SO-H, - S02N H 2 ,
`5-tetrazol yl;
`
`R~,is H;
`
`R~„R<,„ and R7, are each independently selected from the < roup consistin< of
`
`H, -OH, oxo, halo <~en, -Ci <alkyl, -O-Ci<alkyl, -S-Ci <alkyl, -NRxxR>x, -ORi2x, -Het2
`
`and -Ar2; each of Ci <alkyl bein <~ optionally substituted with from 1 to 3 substituents
`
`selected from -OH and halo < en;
`
`Rx„R > „ a n d Ri 2x are each independently selected from the < roup consistin< of
`
`H, -OH, halo, -Ci <,alkyl, -0-Ci <,alkyl, -S-Ci <,alkyl, and -Ar;;
`
`Rni„R i i „ R i ; x and Ri<x are each independently selected from the < roup
`
`consistin<~ of H, -OH, halo <~en, -Ci <alkyl, -0-Ci <alkyl, and -S-Ci <alkyl; Ari, Ar2 and
`
`Ar; are each independently a 5- or 6-membered aromatic monocycle optionally
`
`comprisin <~ 1 or 2 heteroatoms selected from 0, N and S; each of Ari, Ar2 and Ar;
`
`bein<~ optionally and independently substituted with from 1 to 3 substituents selected
`
`from - N R n >xRi ix, -Ci <alkyl, -0-Ci <alkyl, and -S-Ci <alkyl;
`
`Het2 is a 5- or 6-membered non-aromatic monocycle optionally comprisin <~ 1
`
`or 2 heteroatoms selected from 0, N and S; Het2 bein < optionally substituted with
`
`from 1 to 3 substituents selected from -NR i ; xRi<„ - C i <alkyl,
`c,alkyl;
`
`-O-C)-(alkyl, and -S-Ci
`
`v is 0, 1, 2, or 3; and
`
`10
`
`Petitioner GE Healthcare – Ex. 1002, p. 13
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`

`

`Attorney Docket: JHU-36631.303
`
`represents a 5 to 10-membered N-containin <~ aromatic or non-aromatic mono(cid:173)
`
`or bicyclic heterocycle, said heterocycle optionally further comprisin <~ 1, 2 or 3
`
`heteroatoms selected from 0, N and S.
`
`In particular