WO2013039851A1 - Optical agents for imaging and visualization of matrix metalloproteinase enzymes - Google Patents

Optical agents for imaging and visualization of matrix metalloproteinase enzymes Download PDF

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Publication number
WO2013039851A1
WO2013039851A1 PCT/US2012/054550 US2012054550W WO2013039851A1 WO 2013039851 A1 WO2013039851 A1 WO 2013039851A1 US 2012054550 W US2012054550 W US 2012054550W WO 2013039851 A1 WO2013039851 A1 WO 2013039851A1
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compound
formula
cor
cancer
compounds
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PCT/US2012/054550
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French (fr)
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John N. Freskos
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Mallinckrodt Llc
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
    • C07D401/06—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings linked by a carbon chain containing only aliphatic carbon atoms
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
    • C07D401/08—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings linked by a carbon chain containing alicyclic rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
    • C07D401/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings linked by a chain containing hetero atoms as chain links

Definitions

  • Matrix metalloproteinases are a class of zinc-dependent endopeptidases having functionality for degradation and remodeling of the extracellular matrix and subendothelial basement membrane. This family of more than 20 enzymes is further classified in several main groups, including gelatinases, interstial collagenases, stromelysins, and membrane-type matrix metalloproteinases, and is typically referenced using the abbreviation "MMP,” optionally followed by a number providing differentiation between specific enzymes of the family (e.g., MMP-1 , MMP-2, etc.).
  • MMP matrix metalloproteinases
  • MMPs are capable of degrading all components of the extracellular matrix, including fibrallar and non-fibrallar collagens, fibronectin, laminin, and basement membrane glycoproteins.
  • the biological activity of MMPs is regulated on a number of levels.
  • MMP genes are transcriptionally responsive to a variety of oncogenes, growth factors, cytokins and hormones.
  • MMP activity is regulated by the combination of secretion, enzyme mediated activation and inhibition via naturally occurring MMP inhibitors such as a 2 - macroglobulin and a variety of tissue inhibitors of metalloproteinases (TIMPs).
  • TMPs tissue inhibitors of metalloproteinases
  • MPPs are believed to play a role in a number of physiological and pathological processes involved in the onset and progression of certain diseases characterized by breakdown of connective tissue and/or extracellular matrix.
  • Pathological conditions for which MMPs are thought to be implicated include: rheumatoid arthritis; osteoarthritis; septic arthritis, corneal, epidermal and gastric ulceration; tumor metastasis, invasion and angiogenesis;
  • MMP-2 and MMP-9 are frequently coexpressed in a number of human cancers, such as colorectal carcinomas, breast cancer and nonsmall cell lung cancers.
  • MMPs play a critical role in establishing and maintaining a macroenvironment that facilitates growth and angiogenesis of tumors at primary and metastatic sites.
  • MMPs are believed to be involved with biological processes associated with inflammation, cardiovascular disease and acute and chronic neuropathic pain states. Elevated serum levels of MMP-3, for example, have been observed in rheumatoid arthritis patients and have also been shown to correlate with systemic markers of inflammation under some conditions.
  • Serum levels of collagenase, stromelysin-1 , and TIMP-1 Serum levels of collagenase, stromelysin-1 , and TIMP-1 .
  • MMPs inhibitors in development and in various stages of clinical trials include broad spectrum inhibitors capable of simultaneously blocking multiple MMPs.
  • the broad spectrum MMP inhibitor marmastat for example, has been reported as effective for the treatment of gastric cancer and pancreatic cancer.
  • Prinomastat is another broad spectrum MMP inhibitor in advanced clinical trials for the treatment of cancer.
  • Selective MMP inhibitors are also in development that are capable of blocking activity of a selected MMP or group of MMPs associated with specific pathological condition.
  • a motivation for development of selective MMP inhibitors is enhancing efficacy for certain therapeutic applications and reducing unwanted side effects associated with blocking the activity of certain MMPs.
  • Aranapakam et al. describe a series of a-sulfone hydroxamates developed in the context of treating osteoarthritis that are alleged to exhibit activity for MMP inhibition.
  • MMPs are also of potential interest as diagnostic agents.
  • a number of studies demonstrate the potential for various classes of MMPs to provide biomarkers for identifying the onset and the prognosis of certain cancers.
  • Expression of stromelysin-3 has been observed in invasive breast carcinomas and is not observed in normal breast tissue, thereby supporting a potential diagnostic application of this class of MMPs.
  • Stromelysin-3 An independent prognostic factor for relapse-free survival in node-positive breast cancer and demonstration of novel breast carcinoma cell expression.
  • matrilysin has also been suggested to be of prognostic value in evaluation of colon and esophageal cancers.
  • MMP 7 matrilysin
  • Ichikawa, Y. et al. Detection of regional lymph node metastases in colon cancer by using RT-PCT for matrix metalloproteinase-7, matrilysin; Clin. Exp. Metastasis, 16:3- 8, 1998; and Yamamoto, H. et al., Association of matrilysin expression with recurrence and poor prognosis in human esophageal squamous cell carcinoma,; Cancer Res. 59:3313-3316, 1999.]
  • MMPs are a well recognized target for development of therapeutic and diagnostic agents, their potential for biomedical imaging and visualization remains much less explored.
  • Development of optical agents for imaging expression of MMPs in certain tumors and lesions has potential application for diagnosis, staging and monitoring of cancer and for the evaluation of tissue degradation associated with pathological conditions including heart disease and arthritis.
  • optical agents capable of selective delivery to cells and tissue expressing MMPs also has potential for visually differentiating the tissue of a tumor or lesion from normal tissue during a surgical procedure, such as a biopsy or therapeutic tissue removal procedure.
  • optical agents preferably have large extinction coefficients for visible and near infrared (NIR) electromagnetic radiation having wavelengths from about 350 nm to about 900 nm, and for fluorescent optical agents optionally have fluorescence quantum yields and Stokes shifts useful for fluorescence imaging and detection.
  • NIR visible and near infrared
  • fluorescent optical agents optionally have fluorescence quantum yields and Stokes shifts useful for fluorescence imaging and detection.
  • optical agents for these applications preferably exhibit low systemic toxicity, low mutagenicity, and rapid clearance from the blood stream.
  • optical agents for these applications are preferably compatible with effective formulation and administration to a target tissue expressing MMPs, for example by exhibiting a reasonable solubility in aqueous solution, a low tendency for aggregation in solution and/or compatibility with pharmaceutical delivery systems, such as liposomes or micelle delivery agents.
  • optical agents for imaging and visualization for diagnosis, evaluation and treatment of disorders associated with expression of MMPs.
  • optical agents for imaging and visualization are needed exhibiting specificity for important target tissues expressing MMPs, such as tumors and other lesions.
  • optical agents are needed having optical, physiochemical and pharmacokinetic properties providing for useful administration, delivery and excitation with electromagnetic radiation.
  • the invention relates generally to optical agents for biomedical applications including imaging, visualization, phototherapy and diagnostic monitoring of cells and tissues expressing MMPs and/or tissues associated with the expression of MMPs.
  • optical agents of the present invention selectively bind to, or otherwise preferentially associate with, a target tissue expressing one or more MMPs, and function to couple
  • optical agents enable a versatile diagnostic platform useful for in vivo, in vitro and ex vivo diagnostic monitoring, visualization and imaging applications, such as, but not limited to, tomographic, photoacoustic and/or sonofluorescent imaging; monitoring and evaluating organ functioning; anatomical visualization; coronary angiography; and fluorescence endoscopy.
  • the invention provides tumor-specific optical agents for detection, diagnosis and/or imaging of cancer.
  • optical agents of the present invention provide tandem imaging and therapeutic functionality, including phototherapy, for example by simultaneously functioning as a MMP inhibitor, an optical probe and/or a phototherapeutic agent, such as a Type 1 or Type 2 phototherapy agent.
  • optical agents including compositions, preparations and formulations, and methods of using and making optical agents for diagnostic and clinical applications.
  • Optical agents of the invention include compounds having a sulfone hydroxamic acid or hydroxamate backbone component providing molecular recognition and/or targeting functionality for one or more MMPs and/or tissues expressing or associated with expression of MMPs and an optical dye component providing preselected optical properties, such as absorption and/or emission in the visible and/or near infrared region of the electromagnetic spectrum (e.g., 350 nm - 1300 nm or preferably for some applications 400 nm to 900 nm).
  • an optical agent of the invention comprises a sulfone hydroxamic acid or hydroxamate derivative including a pyrazine, cyanine, indocyanine, azulene or azaazulene component having ring substituents providing large absorption in the visible and/or near infrared region of the electromagnetic spectrum, and optionally a fluorescence quantum yield and Stokes shift useful for optical excitation and detection in biomedical procedures.
  • embodiments of the present invention provide exogenous optical agents for biomedical and bioanalytical applications including imaging, visualization, diagnostic monitoring, and therapy.
  • the invention provides compounds being of formula (FX1 ) or (FX2):
  • each ring Z is independently
  • Compounds of the invention for example compounds having formula (FX1 ) - (FX113),
  • the invention provides compounds being of the formula (FX3) or (FX4):
  • optical dye refers to a functional group of a compound that absorbs, emits, and/or scatters electromagnetic radiation in the visible and/or infrared regions of the
  • electromagnetic spectrum for example, electromagnetic radiation having wavelengths selected over the range of 350 - 1300 nanometers, and optionally for some applications over the range of 400 nm to 900 nm.
  • Optical dyes of some compounds of the invention function as a chromophore, fluorophore and/or functional group exhibiting photoacoustic properties, for example, when provided in an in vivo environment.
  • Optical dyes of some compounds of the invention enable the compound to undergo emission processes upon absorption of
  • electromagnetic radiation for example via fluorescence and/or phosphorescence pathways.
  • Optical dyes of some compounds of the invention enable the compound to undergo
  • fluorescence of visible or infrared electromagnetic radiation for example, via emission of electromagnetic radiation having wavelengths selected over the range of 350 - 1300 nanometers, and optionally for some applications over the range of 400 nm to 900 nm.
  • Optical dyes of some compounds of the invention enable the compound to undergo fluorescence of visible or infrared electromagnetic radiation, for example, by exhibiting a fluorescence quantum yield in an in vivo environment that is greater than or equal to 0.0001 , optionally for some embodiments greater than or equal to 0.001 , optionally for some embodiments greater than or equal to 0.01 , and optionally for some embodiments greater than or equal to 0.1.
  • Optical dyes of some compounds of the invention enable the compound to undergo fluorescence of visible or infrared electromagnetic radiation, for example, by exhibiting a Stokes shift in an in vivo environment greater than or equal to 5 nanometers, optionally for some embodiments greater than or equal to 10 nanometers and optionally for some embodiments greater than or equal to 20 nanometers.
  • Optical dyes in some compounds are directly or indirectly covalently linked to the sulfone hydroxamic acid or hydroxamate backbone of the compound, for example, by linking groups and/or spacing groups that do not interfere with the molecular recognition and/or targeting functionality of the hydroxamic acid or hydroxamate backbone with respect to MMPs and/or tissues associate with MMPs.
  • the invention provides compounds having any of formula (FX1 ) - (FX27), wherein A is a group corresponding to a pyrazine, a cyanine, an indocyanine, phthalocyanine, a rhodamine, a thiazole, a phenylxanthene, a phenothiazine, a
  • phenoselenazine a squaraine, a dipyrrolo pyrimidone, an anthraquinone, a tetracene, a quinoline, an acridine, an acridone, a phenanthridine, an azo dye, a phenoxazine, an azulene, an azaazulene, a triphenyl methane dye, an indole, a benzoindole, an indocarbocyanine, a Nile Red dye, a thionin dye, an isosulfan blue dye, or a benzoindocarbocyanine, including for example, derivatives thereof having one or more electron withdrawing groups, electron donating groups and/or photosensitizer groups.
  • a compound having any of formula (FX1 ) - (FX27), is provided wherein A is a group corresponding to a pyrazine, a cyanine, azulene, azaazulene or an indocyanine, including for example, derivatives thereof having one or more electron withdrawing groups, electron donating groups and/or
  • reference to embodiments wherein q, x and/or r is equal to 0 refers to compounds where L 1 , L 2 or L 3 , respectively, is not present and reference to embodiments wherein q, x and/or r is equal to 1 refers to compounds where L 1 , L 2 or L 3 , respectively, is present.
  • W 2 is directly linked to ring Z when x is equal to 0 and W 3 is directly linked to ring Z when r is equal to 0.
  • W 1 is directly linked to an adjacent piperidine group when q is equal to 0.
  • Embodiments wherein W 2 is a single bond and x is equal to 0 refer to compositions having a phenyl group or piperidine group (optionally substituted) directly bonded to ring Z and embodiments wherein W 3 is a single bond and r is equal to 0 refer to formula having an optical dye (optionally substituted) directed bonded to ring Z.
  • W 1 is a single bond and q is equal to 0 refer to compositions having an optical dye (optionally substituted) directed bonded to an adjacent piperidine group.
  • a group corresponding to an indicated species expressly includes a radical of the species or group, such as an aromatic radical or heterocyclic aromatic radical of the species or group of species provided in a covalently bonded configuration, optionally with one or more substituents, including but not limited to electron donating groups, electron withdrawing groups, photosensitizers and/or targeting ligands.
  • the sulfone hydroxamic acid backbone of some compounds having formula (FX1 ) or (FX2) provides selective binding with MMPs and the optical dye component provides useful optical functionality.
  • Optical agents of the present invention providing the optical dye component in the solvent exposed portion of the MMP binding pocket or in the SV MMP binding pocket are particularly useful for some applications, as binding of the optical agent to a MMP may generate an observable change in the optical properties of the optical agent, such as an observable change in the excitation or emission wavelengths, fluorescence quantum yield or Stokes shift, which may be useful for enhancing optical detection, imaging or monitoring cells and/or tissue expressing MMPs or associate with MMP expression.
  • the invention provides compounds being of the formula (FX5), (FX6), (FX7) or (FX8):
  • the invention provides compounds being of the formula (FX9), (FX10),
  • the invention provides compounds being of the formula (FX13) or (FX14): eutically acceptable salt or ester thereof; wherein: B is -O- or -S- and wherein Ring Z, L 1 , W 1 , A, R 1 to R 5 , q, and U are as set forth in the description of formula (FX1 ) and (FX2).
  • the invention provides compounds being of the formula (FX15) or (FX16):
  • Ring Z, L 1 , W 1 , A, R 1 to R 5 , q, and U are as set forth in the description of formula (FX1 ) and (FX2).
  • the invention provides compounds being of the formula (FX17), (FX18) or (FX19): (FX19); or a pharmaceutically acceptable salt or ester thereof: wherein Ring Z, L 3 , W 3 , A, R 10 , r, and U are as set forth in the description of formula (FX1 ) and (FX2).
  • the invention provides compounds being of the formula (FX20), (FX21 ), (FX22) or (FX23):
  • the invention provides compounds being of the formula (FX24), (FX25),
  • the invention provides compounds having any one of formula (FX1 ) - (FX27), wherein A is a group corresponding to a pyrazine, for example a pyrazine providing a chromophore and/or fluorophore that absorbs electromagnetic radiation having wavelengths in the visible and/or infrared regions of the electromagnetic spectrum (e.g., 350 nm to 1300 nm, or optionally 400 nm to 900 nm).
  • Embodiments of the invention include, for example, compounds having any one of formula (FX1 ) - (FX27), wherein A is a group corresponding to a pyrazine having one or more electron donating groups, electron withdrawing groups and/or
  • the invention provides compounds being of the formula (FX28) or (FX29):
  • each of X 1 , X 2 and X 3 is independently hydrogen, - OCF 3 , C C 6 alkyl, C 3 -C 6 cycloalkyl, C 5 -C 10 aryl, C 5 -C 10 heteroaryl, C C 6 acyl, C 2 -C 6 alkenyl, C 2 - C 6 alkynyl, C 5 -C 10 alkylaryl, halo, halomethyl, dihalomethyl, trihalomethyl, -C0 2 R 4 °, -SOR 41 , - OSR 42 , -S0 2 OR 43 , -CH 2 (CH 2 OCH 2 ) c CH 2 OH, -P0 3 R 44 R 45 , -OR 46 , -SR 47 , -NR 48 R 49 , - NR 50 COR 51 , -CN, -CONR 52 R 53 , -COR 54 , -N
  • the invention provides compounds being of the formula (FX30), (FX31 ),
  • the invention provides compounds being of the formula (FX34), (FX35), (FX36) or (FX37):
  • the invention provides compounds being of the formula (FX38) or (FX39):
  • the invention provides compounds being of the formula (FX40) or (FX41 ):
  • the invention provides compounds being of the formula (FX42), (FX43) or (FX44):
  • the invention provides compounds being of the formula (FX45), (FX46), (FX47) or (FX48):
  • the invention provides compounds being of the formula (FX49), (FX50), (FX51 ) or (FX52): (FX52); or a pharmaceutically acceptable salt or ester thereof; wherein: m, n, X 1 to X 3 , R 10 , R 48 , R 49 , and R 54 are as set forth in the description of formula (FX1 ), (FX2), (FX28) and (FX29).
  • the invention provides compounds being of the formula (FX53), (FX54), (FX55) or (FX56):
  • B is -O- or -S-; wherein ring Z, a, m, n, X 1 to X 3 , R 1 to R 5 are as set forth in the description of formula (FX1), (FX2), (FX28) and (FX29).
  • the invention provides compounds being of the formula (FX57), (FX58), or
  • the invention provides compounds being of the formula (FX60), (FX61 ), or
  • R 48 R 49 (FX62); or a pharmaceutically acceptable salt or ester thereof; wherein: R 48 and R 49 are as set forth in the description of formula (FX1 ), (FX2), (FX28) and (FX29).
  • the invention provides compounds having any one of formula (FX1 ) - (FX27), wherein A is a group corresponding to an indocyanine, for example an indocyanine providing a chromophore and/or fluorophore that absorbs electromagnetic radiation having wavelengths in the visible and/or infrared regions of the electromagnetic spectrum (e.g., 350 nm to 1300 nm, or optionally 400 nm to 900 nm).
  • A is a group corresponding to an indocyanine, for example an indocyanine providing a chromophore and/or fluorophore that absorbs electromagnetic radiation having wavelengths in the visible and/or infrared regions of the electromagnetic spectrum (e.g., 350 nm to 1300 nm, or optionally 400 nm to 900 nm).
  • Embodiments of the invention include, for example, compounds having any one of formula (FX1 ) - (FX27), wherein A is a group corresponding to an indocyanine having one or more electron donating groups, electron withdrawing groups and/or photosensitizer groups as substituents.
  • the invention provides compounds being of the formula (FX63) or (FX64):
  • each of X 1 to X 17 is independently hydrogen, -OCF 3 , Ci-C 6 alkyl, C 3 -C 6 cycloalkyl, C5-C10 aryl, C5-C10 heteroaryl, CrC 6 acyl, C 2 -C 6 alkenyl, C 2 -C 6 alkynyl, C5-C 10 alkylaryl, halo, halomethyl, dihalomethyl, trihalomethyl, -C0 2 R 4 °, -SOR 41 , -OSR 42 , - SO 2 OR 43 , -CH 2 (CH 2 0CH 2 ) c CH 2 OH, -POaR ⁇ R 45 , -OR 46 , -SR 47 , -NR 48 R 49 , -NR 50 COR 51 , -CN, -CONR 52 R 53 , -COR 54 , -NO 2 , -SO
  • the invention provides compounds being of the formula (FX65), (FX66), (FX67) or (FX68):
  • the invention provides compounds being of the formula (FX73) or (FX74):
  • L 1 to L 3 , W 1 to W 3 , X 1 to X 17 , U, j, i, q, r, and x are as set forth in the description of formula (FX1 ), (FX2), (FX63) and (FX64).
  • the invention provides compounds having any one of formula (FX1 ) - (FX27), wherein A is a group corresponding to an azulene or azaazulene, for example an azulene or azaazulene providing a chromophore and/or fluorophore that absorbs
  • electromagnetic radiation having wavelengths in the visible and/or infrared regions of the electromagnetic spectrum (e.g., 350 nm to 1300 nm, or optionally 400 nm to 900 nm).
  • Embodiments of the invention include, for example, compounds having any one of formula
  • G 1 is -N-, -C(K)-, or -C(X 1 )-
  • G 2 is -N-, -C(K)-, or - C(X 2 )-
  • G 3 is -N-, -C(K)-, or -C(X 3 )-
  • G 4 is -N-, -C(K)-, or -C(X 4 )-
  • G 5 is -N-, -C(K)-, or - C(X 5 )-
  • G 6 is -N-, -C(K)-, or -C(X 6 )-
  • G 7 is -N-, -C(K)-, or -C(X 7 )-
  • G 8 is -N-, -C(K)-, or - C(X 8 )-; wherein at most one of G 1 - G 8 is -N-; and
  • -C(K)- indicates a direct or indirect linkage between an intra-ring carbon in the azulene or azaazulene group and the backbone of the compound.
  • the -C(K)- linkage is direct wherein the azulene or azaazulene group is directly bonded to ring Z (e.g., a phenyl or piperidine group) or a piperidine of the backbone.
  • the -C(K)- linkage is indirect wherein the azulene group or azaazulene group is bonded to W 1 -W 3 .
  • the -C(K)- linkage is indirect wherein the azulene group or azaazulene group is bonded to L 1 -L 3 .
  • K is a single bond to W 1 , W 2 , or W 3 which is also designated as a single bond refers to direct linking of the azulene or azaazulene group to L 1 , L 2 , or L 3 , if present, or direct linking of the azulene or azaazulene group to the piperidine group or ring Z of the backbone.
  • the invention provides compounds being of the formula being of the formula (FX79) or (FX80):
  • the invention provides compounds being of the formula being of the formula (FX81 ) or (FX82):
  • the invention provides compounds being of the formula (FX83) or (FX84):
  • the invention provides compounds being of the formula (FX85), (FX86), (FX87) or (FX88):
  • the invention provides compounds being of the formula (FX89), (FX90), (FX91 )
  • the invention provides compounds being of the formula (FX93), (FX94), (FX95)
  • L 1 to L 3 , W 1 to W 3 , U, ring Z, G 1 to G 8 , R 1 - R 5 , x, r and q are as described in formulas (FX1 ), (FX2), (FX75), (FX76), (FX77) and (FX78).
  • the invention provides compounds being of the formula (FX97), (FX98),
  • the invention provides compounds being of the formula (FX101 ), (FX102), (FX103) or (FX104):
  • the present invention includes therapeutic agents useful for biomedical applications comprising purified stereoisomers (e.g., enantiomers and diastereomers), salts (including quarternary salts), and/or ionic forms (e.g., protonated and deprotonated forms) of the compounds of any of formula (FX1 ) - (FX113) , and mixtures thereof.
  • purified stereoisomers e.g., enantiomers and diastereomers
  • salts including quarternary salts
  • ionic forms e.g., protonated and deprotonated forms
  • acidic functional groups and basic functional groups of the compounds of any of formula (FX1 ) - (FX113) may be in protonated or deprotonated states depending on the molecular environment (e.g., pH, ionic strength, composition, etc.), for example during synthesis, formulation and/or administration.
  • the invention includes, but is not limited to, the hydroxamate forms, and related salts, of all of the hydroxamic acids shown in formula (FX1 ) - (FX113).
  • compositions having formula (FX1) - (FX113) is selected to achieve preselected properties, such as optical, physiochemical and pharmacokinetic properties useful for biomedical applications.
  • the invention provides, for example, compositions having any one of (FX1 ) - (FX113) wherein at least one of X 1 to X 3 or X 1 to X 17 X 1 to X 8 is an electron
  • EWG electron withdrawing group
  • Incorporation of a combination of an EWG and an EDG as substituents of different carbon atoms of the pyrazine group or indocyanine group or azulene group or azaazulene group is particularly beneficial for providing optical agents having large extinction coefficients in the visible and near infrared regions of the electromagnetic spectrum (e.g., 350 nm - 1300 nm, optionally 400 nm to 900 nm), emission in the visible and near infrared regions(e.g., 350 nm - 1300 nm, optionally 500 - 900 nm), a large fluorescence quantum yield (e.g., >0.1 ) and a Stoke's shift useful for optical detection and imaging (e.g., Stoke's shift > 10 nm).
  • optical agents having large extinction coefficients in the visible and near infrared regions of the electromagnetic spectrum (e.g., 350 nm - 1300 nm, optionally 400 nm to 900 nm),
  • an electron withdrawing group and electron donating group are positioned on adjacent carbon atoms of the pyrazine group or indocyanine group or azulene group or azaazulene group.
  • the invention includes embodiments wherein an electron withdrawing group and an electron donating group are positioned on non-adjacent carbon atoms of the pyrazine group or indocyanine group or azulene group or azaazulene group.
  • Multiple electron withdrawing groups and/or electron donating groups on each substituent arm of the pyrazine group or indocyanine group or azulene group or azaazulene group are contemplated by the compositions of this aspect of the invention.
  • one EWG arm may comprise two, three, or more electron withdrawing groups bonded to the pyrazine or indocyanine or azulene group or azaazulene group core via a common linking moiety.
  • the invention provides a compound having any one of formula (FX1 ) - (FX113), wherein at least one of X 1 to X 3 or X 1 to X 17 or X 1 to X 8 is an electron donating group, and wherein at least one of X 1 to X 3 or X 1 to X 17 or X 1 to X 8 is an electron withdrawing group.
  • the invention provides a compound having any one of formula (FX1 ) - (FX113), wherein at least one of X 1 to X 3 or X 1 to X 17 or X 1 to X 8 is Ci-C 6 alkyl, -OR 46 , -SR 47 , -NR 48 R 49 , or -NR 50 COR 51 ; and wherein at least one of X 1 to X 3 or X 1 to X 17 or X 1 to X 8 is -CN, -C0 2 R 4 °, -S0 2 OR 43 , -CONR 52 R 53 , -COR 54 , -N0 2 , -SOR 41 , -S0 2 R 55 , -P0 3 R 44 R 45 , halo, C C 6 acyl, trihalomethyl, or -S0 2 NR 56 R 57 .
  • formula (FX1 ) - (FX113) wherein at least one of X 1 to X
  • the invention provides a compound having any one of formula (FX1 ) - (FX113), wherein at least one of X 1 to X 3 or X 1 to X 17 or X 1 to X 8 is -OR 46 , -SR 47 , -NR 48 R 49 , or -NR 50 COR 51 .
  • the invention provides a compound having any one of formula (FX1 ) - (FX113), wherein at least one of X 1 to X 3 or X 1 to X 17 or X 1 to X 8 is -NR 48 R 49 , or -NR 50 COR 51 .
  • the invention provides a compound having any one of formula (FX1 ) - (FX113), wherein at least one of X 1 to X 3 or X 1 to X 17 or X 1 to X 8 is -CN, -C0 2 R 40 ,- COR 54 , -N0 2 , -S0 2 R 55 , or -S0 2 NR 56 R 57 .
  • the invention provides a compound having any one of formula (FX1 ) - (FX113), wherein at least one of X 1 to X 3 or X 1 to X 17 or X 1 to X 8 is -C0 2 R 40 , -COR 54 , -S0 2 NR 56 R 57 or -S0 2 R 55 .
  • the invention provides a compound having any one of formula (FX1 ) - (FX113), wherein at least one of X 1 to X 3 or X 1 to X 17 or X 1 to X 8 is -NR 48 R 49 , and wherein at least one of X 1 to X 3 or X 1 to X 17 or X 1 to X 8 is -CO 2 R 40 ,-COR 54 , -S0 2 NR 56 R 57 or -S0 2 R 55 .
  • the present invention provides compositions having any one of formula (FX28) - (FX62), wherein at least one of X 1 , X 2 and X 3 is -OR 46 , -SR 47 , -NR 48 R 49 , and - NR 50 COR 51 .
  • the present invention provides compositions having any one of formula (FX28) - (FX62), wherein at least one of X 1 , X 2 and X 3 is -NR 48 R 49 , or -NR 50 COR 51 .
  • the present invention provides compositions having any one of formula (FX28) - (FX62), wherein at least one of X 1 , X 2 and X 3 is -CN, -CO ⁇ -COR 54 , -N0 2 , -S0 2 R 55 , or— S0 2 NR 56 R 57 .
  • the present invention provides compositions having any one of formula (FX28) - (FX62), wherein at least one of X 1 , X 2 and X 3 is -COzR ⁇ -COR 54 ,—
  • the invention provides compounds with electron-donating and electron-withdrawing groups attached to adjacent positions of a pyrazine optical dye component.
  • any one of X 1 and X 2 is C C 6 alkyl, -OR 46 , -SR 47 , -NR 48 R 49 , or -NR 50 COR 51 and the other of X 1 and X 2 is -CN, -C0 2 R 40 , -S0 2 OR 43 , -CONR 52 R 53 , -COR 54 , -N0 2 , -SOR 41 , -S0 2 R 55 , - POsR ⁇ R 45 , halo, C C 6 acyl, trihalomethyl, or -S0 2 NR 56 R 57 ;
  • any one of X 1 and X 3 is C C 6 alkyl, -OR 46 , -SR 47 , -NR 48 R 49 , or -NR 50 COR 51 and the other of X 1 and X 3 is -CN, -C0 2 R 40 , -S0 2 OR 43 , -CONR 52 R 53 , -COR 54 , -N0 2 , -SOR 41 , -S0 2 R 55 , - POsR ⁇ R 45 , halo, C C 6 acyl, trihalomethyl, or -S0 2 NR 56 R 57 ;
  • any one of X 2 and X 3 is C C 6 alkyl, -OR 46 , -SR 47 , -NR 48 R 49 , or -NR 50 COR 51 and the other of X 2 and X 3 is -CN, -C0 2 R 40 , -S0 2 OR 43 , -CONR 52 R 53 , -COR 54 , -N0 2 , -SOR 41 , -S0 2 R 55 , - POsR ⁇ R 45 , halo, C C 6 acyl, trihalomethyl, or -S0 2 NR 56 R 57 ;
  • any two of X 1 , X 2 and X 3 is C C 6 alkyl, -OR 46 , -SR 47 , -NR 48 R 49 , or -NR 50 COR 51 and the other of X 1 , X 2 and X 3 is -CN, -C0 2 R 40 , -S0 2 OR 43 , -CONR 52 R 53 , -COR 54 , -N0 2 , -SOR 41 , - S0 2 R 55 , -POsR ⁇ R 45 , halo, C C 6 acyl, trihalomethyl, or -S0 2 NR 56 R 57 ; or
  • any two of X 1 , X 2 and X 3 is -CN, -C0 2 R 40 , -S0 2 OR 43 , -CONR 52 R 53 , -COR 54 , -N0 2 , -SOR 41 , -S0 2 R 55 , -P0 3 R 44 R 45 , halo, C C 6 acyl, trihalomethyl, or -S0 2 NR 56 R 57 and the other of X 1 , X 2 and X 3 is C C 6 alkyl, -OR 46 , -SR 47 , -NR 48 R 49 , or -NR 50 COR 51 .
  • a compound of the present invention has any one of formula (FX1 ) - (FX113), wherein each of R 1 and R 5 is hydrogen. In an embodiment, a compound of the present invention has any one of formula (FX1 ) - (FX113), wherein each of R 1 , R 2 , R 4 and R 5 is hydrogen. In an embodiment, a compound of the present invention has any one of formula
  • a compound of the present invention has any one of formula (FX1 ) - (FX113), wherein R 3 is a group other than hydrogen. In an embodiment, a compound of the present invention has any one of formula
  • a compound of the present invention has any one of formula (FX1) - (FX113), wherein R 3 is PS 1 or PS 2 , and optionally each of R 1 - R 2 and R 4 - R 5 is hydrogen.
  • a compound of the present invention has any one of formula (FX1 ) - (FX113) wherein at least one of X 1 to X 3 or X 1 to X 17 or X 1 to X 8 is PS 1 or PS 2 .
  • linking moieties may be used to covalently link the sulfone hydroxamic acid or hydroxamate backbone and the optical dye component of compounds of the present invention.
  • Some embodiments incorporate an optional spacer moiety (e.g., L 1 , L 2 and/ or L 3 ) between the sulfone hydroxamic acid or hydroxamate backbone and the optical dye component, for example, to ensure that the molecular recognition functionality of the sulfone hydroxamic acid or hydroxamate backbone is retained and/or to preselect or optimize the optical functionality of the optical dye component.
  • the invention provides compounds having any one of formula (FX1 ) - (FX113), wherein at least one of W 1 and L 1 , W 2 and L 2 , or form: -(CH 2 ) n - -(CH 2 ) m NHCO- -CONH(CH 2 ) n CO- , -
  • the invention provides compounds having any one of formula (FX1 ) - (FX113), wherein at least one of W 1 and L 1 , W 2 and L 2 , or W 3 and L 3 combine to form
  • n and a are as set forth in the description of formulas (FX1 ) and (FX2).
  • the invention provides a compound having any of formula (FX1 ) - (FX113), wherein each of R 1 to R 5 and R 8 to R 10 is hydrogen, Ci - C 6 alkyl or -OCF 3 , and optionally for some embodiments wherein each of R 1 to R 5 and R 8 to R 10 is hydrogen or Ci - C 6 alkyl, and optionally for some embodiments wherein each of R 1 to R 5 and R 8 to R 10 is hydrogen or Ci - C 3 alkyl, and optionally for some embodiments wherein each of R 1 to R 5 and R 8 to R 10 is hydrogen.
  • the invention provides a compound having any of formula (FX1 ) - (FX113), wherein at least one of R 1 to R 5 and R 8 to R 10 is -OCF 3 , and optionally wherein R 3 is - OCF 3 .
  • the invention provides a compound having any of formula (FX1 ) - (FX113), wherein at least one of R 1 to R 5 and R 8 to R 10 is PS 1 .
  • the invention provides a compound having any of formula (FX1 ) - (FX113), wherein at least one of R 1 to R 5 and R 8 to R 10 is PS 2 .
  • the invention provides a compound having any of formula (FX1 ) - (FX113), wherein R 1 is hydrogen; and/or R 2 is hydrogen; and/or R 4 is hydrogen; and/or R 5 is hydrogen.
  • the invention provides a compound having any of formula (FX1 ) - (FX113), wherein U is -O- or -S- and optionally for some embodiments wherein U is -0-.
  • the invention provides a compound having any of formula (FX1 ) - (FX113), wherein each of R 11 to R 34 and R 60 to R 79 is independently hydrogen or CrC 6 alkyl; and optionally wherein each of R 11 to R 34 and R 60 to R 79 is
  • the invention provides a compound having any of formula (FX63) - (FX74), wherein each of X 1 to X 17 is hydrogen, an electron donating group or an electron withdrawing group, and optionally wherein at least two of X 1 to X 17 is an electron donating group or an electron withdrawing group and the rest of X 1 to X 17 is hydrogen.
  • the invention provides a compound having any of formula (FX28) - (FX62), wherein each of X 1 to X 3 is hydrogen, an electron donating group or an electron withdrawing group, and optionally wherein at least two of X 1 to X 3 is an electron donating group or an electron withdrawing group and the rest of X 1 to X 3 is hydrogen.
  • the invention provides a compound having any of formula (FX75) - (FX104), wherein each of X 1 to X 8 is hydrogen, an electron donating group or an electron withdrawing group, and optionally wherein at least two of X 1 to X 8 is an electron donating group or an electron withdrawing group and the rest of X 1 to X 8 is hydrogen.
  • the invention provides a compound having any of formula (FX1 ) - (FX113), wherein each of each of R 40 to R 57 is hydrogen or C-
  • the invention provides a compound having any of formula (FX1 ) - (FX113), wherein Y is -CR 2 -, and optionally wherein Y is -CH-.
  • the invention provides a compound having any of formula (FX1 ) - (FX113), wherein Y is -N-.
  • the invention provides a compound having any of formula (FX1 ) - (FX113), wherein each of h and g are 1 or 2, and optionally wherein each of h and g are 1.
  • a compound of the invention having any of formula (FX1 ) - (FX113) selectively binds to one or more MMPs, preferably for some applications under in vivo conditions.
  • compounds of the present invention are selective with respect to which MMP they bind to or otherwise associate with.
  • the invention provides compounds that bind to a single MMP or a group of selected MMPs, such as MMP-2, MMP-9 and/or MMP-13.
  • the invention provides compounds that bind to some MMPs and do not bind with one or more selected MMPs, such as MMP-1 .
  • the compounds provided are MMP selective compounds.
  • a compound provided has a sulfone hydroxamic acid or hydroxamate backbone that binds selectively to one or more MMPs.
  • the invention provides compounds having any of formula (FX1 ) - (FX113) that inhibit the biological activity of one or more MMPs, for example by inhibiting biological activity associated with an MMP or a class of MMPs involved in degradation and/or remodeling of the extracellular matrix.
  • a compound provided selectively binds to a related metalloproteinase called TNF-a Converting Enzyme (TACE) which is implicated in the cartilage catabolism pathway.
  • TACE TNF-a Converting Enzyme
  • compounds of the invention may optionally include a poly(ethylene glycol) (abbreviated as PEG) component.
  • PEG poly(ethylene glycol)
  • the invention provides a composition having any one of the formula (FX1) - (FX113), wherein at least one of W 1 to W 3 , L 1 to L 3 , R 1 to R 5 and R 8 to R 10 is a substituent comprising -(CH 2 OCH 2 ) m-, or a derivative thereof, wherein m is selected from the range of 1 to 100.
  • Incorporation of a poly(ethylene glycol) component in some compositions of the invention provides
  • Poly(ethylene glycol) containing compounds of some embodiments of the present invention provide enhanced biocompatibility, low toxicity and suppress immune responses upon administration.
  • Poly(ethylene glycol) containing compounds of some embodiments of the invention facilitate formulation, administration and/or delivery, for example, by enhancing solubility.
  • the invention provides a compound being of the formula (FX105),
  • the invention further provides a compound having any one of formula (FX1 ) - (FX113), or a pharmaceutical formulation thereof, for use in an optical imaging, diagnostic, and/or therapeutic biomedical procedure.
  • the invention provides an optical agent comprising a pharmaceutically acceptable formulation, wherein at least one active ingredient of the formulation is a compound having any one of formula (FX1 ) - (FX113) provided in a therapeutically or diagnostically effective amount.
  • the invention includes, for example, formulations comprising a compound having any one of formula (FX1 ) - (FX113) and one or more pharmaceutically acceptable carriers or excipients.
  • the biomedical procedure comprises: (i) administering to a subject a therapeutically or diagnostically effective amount of the compound having any one of formula (FX1 ) - (FX113) under conditions sufficient for contacting a target tissue and/or target cell with the compound, wherein the compound selectively binds to matrix metalloproteinase enzyme expressed by, or otherwise associated with, the target tissue or target cell (e.g., target tissue characterized by an elevated
  • the compound is administered to the subject in an amount sufficient to inhibit the biological activity of MMPs expressed or otherwise generated by the target tissue or target cells.
  • the compound is administered to the subject in an amount sufficient to suppress or otherwise attenuate biological processes associated with MMPs in the target tissue, such as the degradation and/or remodeling of the extra cellular matrix.
  • the compound is administered to the tissue of a tumor of the subject in an amount sufficient to suppress or prevent the growth, proliferation and/or metastasis of the tumor.
  • the administered compound is exposed at the site of the target tissue or target cell to electromagnetic radiation having wavelengths selected over a range of 350 nanometers to 1300 nanometers, optionally having wavelengths selected over a range of 350 nanometers to 900 nanometers.
  • exposing the administered compound to electromagnetic radiation generates fluorescence, wherein the biomedical procedure further comprises detecting fluorescence from the administered compound.
  • exposing the administered compound to electromagnetic radiation generates a diagnostically effective amount of fluorescence, for example an amount of fluorescence allowing for optical detection and/or imaging of the target tissue .
  • a method of the invention further comprises exposing the administered compound at the target tissue to electromagnetic radiation having sufficient power, fluence, intensity and/or dose (net number of photons provided to the target tissue) to provide optical detection and/or imagining of the target tissue.
  • a method of the invention further comprises exposing the administered compound at the target tissue to a diagnostically effective and/or therapeutically effective amount of electromagnetic radiation.
  • a method of the invention further comprises generating an image of the fluorescence from the compound.
  • a method of the invention further comprises visualizing the fluorescence from the compound.
  • a method of the invention further comprises measuring the intensity, energy or power of fluorescence from the compound, optionally as a function of illuminated area.
  • the biomedical procedure comprises administering or otherwise targeting the administered compound to a target tissue or target cell of the subject, such as a tumor, lesion, site of inflammation, vasculature tissue, or an organ, wherein the target tissue or target cell expresses or otherwise generates MMPs.
  • a target tissue or target cell of the subject such as a tumor, lesion, site of inflammation, vasculature tissue, or an organ
  • the target tissue or target cell expresses or otherwise generates MMPs.
  • the target tissue is a tissue type selected from the group consisting of colon, prostate, gastric, esophageal, uterine, endometrial, pancreatic, breast, cervical, brain, skin, gallbladder, lung, heart, central nervous system and ovary.
  • Another aspect of the invention is directed to methods of using the compounds of any one of formulas (FX1 ) - (FX113) in biomedical procedures including treatment and diagnosis of diseases and other pathological conditions.
  • the invention includes, for example, methods of using compounds of any one of formulas (FX1 ) - (FX113) in a biomedical procedure, including photodiagnostic and therapeutic methods such as optical imaging, anatomical visualization, endoscopic visualization, image guided surgery, and optical biopsy of tumors and other lesions.
  • the invention includes, for example, methods of using compounds of any one of formulas (FX1 ) - (FX113) in a therapeutic procedure involving inhibition of the biological activity of MMPs in the target tissue.
  • a therapeutically effective amount of a compound of any one of formulas (FX1 ) - (FX113) is administered to a subject (e.g., via intravenous or intraarterial injection, oral administration, topical administration, etc.).
  • a method of the present invention further comprises exposing the administered compound to a subject (e.g., via intravenous or intraarterial injection, oral administration, topical administration, etc.).
  • electromagnetic radiation such as electromagnetic radiation having wavelengths in the visible and near infrared regions of the electromagnetic spectrum (e.g., 350 nm to 1300 nm).
  • electromagnetic radiation exposed to the compound of any one of formulas (FX1 ) - (FX113) does not have wavelengths in the X-ray region of the electromagnetic spectrum.
  • the electromagnetic radiation exposed to the compound of any one of formulas (FX1 ) - (FX113) does not have wavelengths in the ultraviolet region of the electromagnetic spectrum.
  • non-ionizing electromagnetic radiation such as electromagnetic radiation having wavelengths in the visible and near infrared regions of the electromagnetic spectrum (e.g., 350 nm to 1300 nm).
  • Non-ionizing electromagnetic radiation refers to electromagnetic radiation wherein a single photon does not have enough energy to completely remove at least one electron from an atom or molecule of the subject's body.
  • the present invention provides methods for treating cancer or a cancer-related disorder.
  • a method includes administering a therapeutically effective amount of a compound having any one of formulas (FX1 ) - (FX113) to a subject in need of treatment for cancer.
  • these cancers may include colorectal, prostate, gastric, esophageal, uterine-endometrial, pancreatic, breast, cervical, head and neck, hepatic, skin, gallbladder, lung, and ovarian cancers.
  • the present invention provides methods of treating inflammation or inflammation-associated disorders, the method including administering a therapeutically-effective amount of a compound having any one of formulas (FX1 ) - (FX113) to a subject in need of treatment for inflammation or an inflammation- associated disorder.
  • these disorders may include, but are not limited to rheumatoid arthritis; osteoarthritis; and septic arthritis.
  • the invention further provides a compound having any one of formula (FX1 ) - (FX113), or a pharmaceutical formulation thereof, for use in a medical phototherapy procedure, such as a Type 1 or Type 2 phototherapy procedure.
  • a compound of the invention has any one of formula (FX1) - (FX113), wherein at least one of R 1 to R 5 and X 1 to X 3 or X 1 to X 17 or X 1 to X 8 is PS 1 or PS 2 .
  • the invention provides a compound having any one of formula (FX1) - (FX113), wherein at least one of R 1 to R 5 and X 1 to X 3 or X 1 to X 17 or X 1 to X 8 is PS 1 or PS 2 .
  • the invention provides a compound having any one of formula (FX1) - (FX113), or a pharmaceutical formulation thereof, for use in a medical phototherapy procedure, such as a Type 1 or Type 2 phototherapy procedure.
  • a compound of the invention has any one
  • phototherapeutic agent comprising a pharmaceutically acceptable formulation, wherein an active ingredient of the formulation provided in a therapeutically effective amount is a compound having any one of formula (FX1 ) - (FX113).
  • the invention includes, for example, formulations comprising a compound having any one of formula (FX1 ) - (FX113) and one or more
  • phototherapy procedure comprises: (i) administering to a subject in need of treatment a therapeutically effective amount of the compound having any one of formula (FX1 ) - (FX113); and (ii) exposing the administered compound to electromagnetic radiation.
  • the administered compound is exposed to electromagnetic radiation having wavelengths selected over a range of 350 nanometers to 1300 nanometers, optionally having wavelengths selected over a range of 350 nanometers to 900 nanometers.
  • exposing the administered compound to electromagnetic radiation generates one or more radicals, nitrenes, carbenes, ions, and/or singlet oxygen.
  • exposing the administered compound to electromagnetic radiation generates a therapeutically effective amount of photoactivated compound.
  • exposing the administered compound to electromagnetic radiation generates a therapeutically effective amount of reactive species causing localized cell death or injury.
  • the medical phototherapy procedure comprises
  • methods of the invention further comprises exposing the administered compound at the target tissue to light having sufficient power, fluence, intensity and/or dose (net number of photons provided to the target tissue) to result in injury, inactivation and/or death to cells at the target tissue.
  • the invention includes, for example, methods of using the compounds of any one of formulas (FX1 ) - (FX113) in a medical phototherapy procedure.
  • a therapeutically effective amount of a compound of any one of formulas (FX1 ) - (FX113) is administered to a subject (e.g., via intravenous or intraarterial injection, oral administration, topical administration, etc.) and exposed to a therapeutically effective amount of electromagnetic radiation, such as electromagnetic radiation having wavelengths in the visible and near infrared regions of the electromagnetic spectrum (e.g., 3500 nm to 1300 nm).
  • the electromagnetic radiation exposed to the compound of any one of formulas (FX1 ) - (FX113) does not have wavelengths in the X-ray region of the electromagnetic spectrum. In a method, the electromagnetic radiation exposed to the compound of any one of formulas (FX1 ) - (FX113) does not have wavelengths in the ultraviolet region of the electromagnetic spectrum.
  • Figure 1 provides the response curve for inhibition of MMP-2 by compound (FX107). In this Figure, percentage inhibition is plotted as a function of concentration ( ⁇ ).
  • Figure 2 provides the response curve for inhibition of MMP-9 by compound (FX107). In this Figure, percentage inhibition is plotted as a function of concentration ( ⁇ ). [075] Figure 3 provides the response curve for inhibition of MMP-2 by compound (FX108).
  • Figure 4 provides the response curve for inhibition of MMP-2 by compound (FX105).
  • Figure 5 provides the response curve for inhibition of MMP-9 by compound (FX105).
  • Figure 6A provides a high resolution mass spectrum of the compound having formula
  • Figure 6B provides an expanded view of the mass spectrum (top panel) of the compound having formula (FX105) and a simulated mass spectrum (lower panel) for a mass to charge ratio (m/z) range of about 1 149 to about 1 154.
  • Figure 6C provides an expanded view of the mass spectrum (top panel) of the compound having formula (FX105) and a simulated mass spectrum (lower panel) for a mass to charge ratio (m/z) range of about 574 to 576.
  • a composition or compound of the invention is isolated or purified.
  • an isolated or purified compound is at least partially isolated or purified as would be understood in the art.
  • the composition or compound of the invention has a chemical purity of 95%, optionally for some applications 99%, optionally for some applications 99.9%, optionally for some applications 99.99%, and optionally for some applications 99.999% pure.
  • lonizable groups include groups from which a proton can be removed (e.g., -COOH, S0 3 H, etc.) or added (e.g., amines) and groups which can be quaternized (e.g., amines). All possible ionic forms of such molecules and salts thereof are intended to be included individually in the disclosure herein. With regard to salts of the compounds herein, one of ordinary skill in the art can select from among a wide variety of available counterions that are appropriate for preparation of salts of this invention for a given application.
  • indication of the functional group -COOH is intended to also include the deprotonated form of this group, e.g., -COO " .
  • indication of the functional group -SO 3 H is intended to also include the deprotonated form of this group, e.g., -S0 3 " .
  • the compounds of this invention can contain one or more chiral centers. Accordingly, this invention is intended to include racemic mixtures, diasteromers, enantiomers, tautomers and mixtures enriched in one or more stereoisomer.
  • the scope of the invention as described and claimed encompasses the racemic forms of the compounds as well as the individual enantiomers and non-racemic mixtures thereof.
  • group may refer to a functional group of a chemical compound.
  • Groups of the present compounds refer to an atom or a collection of atoms that are a part of the compound.
  • Groups of the present invention may be attached to other atoms of the compound via one or more covalent bonds.
  • Groups may also be characterized with respect to their valence state.
  • the present invention includes groups characterized as monovalent, divalent, trivalent, etc. valence states.
  • -X- refers generally to a bonding configuration wherein X is bonded to two other adjacent atoms and/or functional groups.
  • -X- refers to a linking group or spacer group, including linear and branched groups.
  • -X- refers to an intra-ring member, for example, an intra-ring member of a cycloalkyl group, aryl group, or heteroaryl group.
  • the designation "-X-” does not specify the type of bond (single, double, triple, etc.) that X forms to the adjacent atoms and/or functional groups.
  • the symbol -N- in the context of G 1 -G 8 refers to azaazulene containing- compounds having an intra-ring nitrogen atom bonded to adjacent intra-ring carbon atoms.
  • the symbol -C(X n )-, wherein n is an integer from 1 to 8 in the context of G 1 -G 8 refers to azulene or azaazulene containing- compounds having an intra-ring carbon atom bonded to adjacent intra-ring carbon and/or nitrogen atoms and also bound to substituent X n .
  • alkylene and “alkylene group” are used synonymously and refer to a divalent group derived from an alkyl group as defined herein.
  • the invention includes compounds having one or more alkylene groups. Alkylene groups in some compounds function as attaching and/or spacer groups. Compounds of the invention may have substituted and/or unsubstituted C C 2 o alkylene, CrC 10 alkylene and C C 5 alkylene groups.
  • cycloalkylene and "cycloalkylene group” are used synonymously and refer to a divalent group derived from a cycloalkyl group as defined herein.
  • the invention includes compounds having one or more cycloalkylene groups. Cycloalkyl groups in some compounds function as attaching and/or spacer groups. Compounds of the invention may have substituted and/or unsubstituted C 3 -C 2 o cycloalkylene, C 3 -C 10 cycloalkylene and C 3 -C 5 cycloalkylene groups.
  • arylene and “arylene group” are used synonymously and refer to a divalent group derived from an aryl group as defined herein.
  • the invention includes compounds having one or more arylene groups.
  • an arylene is a divalent group derived from an aryl group by removal of hydrogen atoms from two intra-ring carbon atoms of an aromatic ring of the aryl group.
  • Arylene groups in some compounds function as attaching and/or spacer groups.
  • Arylene groups in some compounds function as chromophore, fluorophore, aromatic antenna, dye and/or imaging groups.
  • Compounds of the invention include substituted and/or unsubstituted C 3 -C 30 arylene, C 3 -C 2 o arylene, C 3 -C 10 arylene and C C 5 arylene groups.
  • heteroarylene and “heteroarylene group” are used synonymously and refer to a divalent group derived from a heteroaryl group as defined herein.
  • the invention includes compounds having one or more heteroarylene groups.
  • a heteroarylene is a divalent group derived from a heteroaryl group by removal of hydrogen atoms from two intra-ring carbon atoms or intra-ring nitrogen atoms of a heteroaromatic or aromatic ring of the heteroaryl group.
  • Heteroarylene groups in some compounds function as attaching and/or spacer groups.
  • Heteroarylene groups in some compounds function as chromophore, aromatic antenna, fluorophore, dye and/or imaging groups.
  • Compounds of the invention include substituted and/or unsubstituted C 3 -C 30 heteroarylene, C 3 -C 2 o heteroarylene, C1-C10 heteroarylene and C 3 -C 5 heteroarylene groups.
  • alkenylene and “alkenylene group” are used synonymously and refer to a divalent group derived from an alkenyl group as defined herein.
  • the invention includes compounds having one or more alkenylene groups. Alkenylene groups in some compounds function as attaching and/or spacer groups. Compounds of the invention include substituted and/or unsubstituted C 2 -C 2 o alkenylene, C 2 -Ci 0 alkenylene and C 2 -C 5 alkenylene groups.
  • cylcoalkenylene and “cylcoalkenylene group” are used synonymously and refer to a divalent group derived from a cylcoalkenyl group as defined herein.
  • the invention includes compounds having one or more cylcoalkenylene groups. Cycloalkenylene groups in some compounds function as attaching and/or spacer groups. Compounds of the invention include substituted and/or unsubstituted C 3 -C 20 cylcoalkenylene, C 3 -C 10 cylcoalkenylene and C 3 -C 5 cylcoalkenylene groups.
  • alkynylene and “alkynylene group” are used synonymously and refer to a divalent group derived from an alkynyl group as defined herein.
  • the invention includes compounds having one or more alkynylene groups. Alkynylene groups in some compounds function as attaching and/or spacer groups. Compounds of the invention include substituted and/or unsubstituted C 2 -C 20 alkynylene, C 2 -C 10 alkynylene and C 2 -C 5 alkynylene groups.
  • halo refers to a halogen group such as a fluoro (-F), chloro (-CI), bromo (-Br), iodo (-I) or astato (-At).
  • heterocyclic refers to ring structures containing at least one other kind of atom, in addition to carbon, in the ring. Examples of such atoms include nitrogen, oxygen and sulfur.
  • heterocyclic rings include, but are not limited to, pyrrolidinyl, piperidyl, imidazolidinyl, tetrahydrofuryl, tetrahydrothienyl, furyl, thienyl, pyridyl, quinolyl, isoquinolyl, pyridazinyl, pyrazinyl, indolyl, imidazolyl, oxazolyl, thiazolyl, pyrazolyl, pyridinyl, benzoxadiazolyl, benzothiadiazolyl, triazolyl and tetrazolyl groups. Atoms of heterocyclic rings can be bonded to a wide range of other atoms and functional groups, for example, provided as substituents.
  • carbocyclic refers to ring structures containing only carbon atoms in the ring. Carbon atoms of carbocyclic rings can be bonded to a wide range of other atoms and functional groups, for example, provided as substituents..
  • alicyclic refers to a ring that is not an aromatic ring. Alicyclic rings include both carbocyclic and heterocyclic rings.
  • alkoxyalkyl refers to a substituent of the formula alkyl-O- alkyl.
  • polyhydroxyalkyl refers to a substituent having from 2 to 12 carbon atoms and from 2 to 5 hydroxyl groups, such as the 2,3-dihydroxypropyl, 2,3,4- trihydroxybutyl or 2,3,4,5-tetrahydroxypentyl residue.
  • polyalkoxyalkyl refers to a substituent of the formula alkyl- (alkoxy) n -alkoxy wherein n is an integer from 1 to 10, preferably 1 to 4, and more preferably for some embodiments 1 to 3.
  • Amino acids include glycine, alanine, valine, leucine, isoleucine, methionine, proline, phenylalanine, tryptophan, asparagine, glutamine, glycine, serine, threonine, serine, Threonine, asparagine, glutamine, tyrosine, cysteine, lysine, arginine, histidine, aspartic acid and glutamic acid.
  • reference to "a side chain residue of a natural a-amino acid” specifically includes the side chains of the above-referenced amino acids.
  • AlkyI groups include straight-chain, branched and cyclic alkyl groups. AlkyI groups include those having from 1 to 30 carbon atoms. Alkyl groups include small alkyl groups having 1 to 3 carbon atoms. Alkyl groups include medium length alkyl groups having from 4-10 carbon atoms. Alkyl groups include long alkyl groups having more than 10 carbon atoms, particularly those having 10-30 carbon atoms.
  • the term cycloalkyl specifically refers to an alky group having a ring structure such as ring structure comprising 3-30 carbon atoms, optionally 3-20 carbon atoms and optionally 2 - 10 carbon atoms, including an alkyl group having one or more rings.
  • Cycloalkyl groups include those having a 3-, 4-, 5-, 6-, 7-, 8-, 9- or 10-member carbon ring(s) and particularly those having a 3-, 4-, 5-, 6-, or 7-member ring(s).
  • the carbon rings in cycloalkyl groups can also carry alkyl groups.
  • Cycloalkyl groups can include bicyclic and tricycloalkyl groups.
  • Alkyl groups are optionally substituted.
  • Substituted alkyl groups include among others those which are substituted with aryl groups, which in turn can be optionally substituted.
  • alkyl groups include methyl, ethyl, n-propyl, iso-propyl, cyclopropyl, n-butyl, s-butyl, t-butyl, cyclobutyl, n-pentyl, branched-pentyl, cyclopentyl, n-hexyl, branched hexyl, and cyclohexyl groups, all of which are optionally substituted.
  • Substituted alkyl groups include fully halogenated or semihalogenated alkyl groups, such as alkyl groups having one or more hydrogens replaced with one or more fluorine atoms, chlorine atoms, bromine atoms and/or iodine atoms.
  • Substituted alkyl groups include fully fluorinated or semifluorinated alkyl groups, such as alkyl groups having one or more hydrogens replaced with one or more fluorine atoms.
  • An alkoxy group is an alkyl group that has been modified by linkage to oxygen and can be represented by the formula R-0 and can also be referred to as an alkyl ether group.
  • alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy, butoxy and heptoxy.
  • Alkoxy groups include substituted alkoxy groups wherein the alky portion of the groups is substituted as provided herein in connection with the description of alkyl groups. As used herein MeO- refers to CH 3 O-.
  • Alkenyl groups include straight-chain, branched and cyclic alkenyl groups. Alkenyl groups include those having 1 , 2 or more double bonds and those in which two or more of the double bonds are conjugated double bonds. Alkenyl groups include those having from 2 to 20 carbon atoms. Alkenyl groups include small alkenyl groups having 2 to 3 carbon atoms. Alkenyl groups include medium length alkenyl groups having from 4-10 carbon atoms. Alkenyl groups include long alkenyl groups having more than 10 carbon atoms, particularly those having 10-20 carbon atoms. Cycloalkenyl groups include those in which a double bond is in the ring or in an alkenyl group attached to a ring.
  • cycloalkenyl specifically refers to an alkenyl group having a ring structure, including an alkenyl group having a 3-, 4-, 5-, 6-, 7-, 8-, 9- or 10-member carbon ring(s) and particularly those having a 3-, 4-, 5-, 6- or 7-member ring(s).
  • the carbon rings in cycloalkenylgroups can also carry alkyl groups.
  • Cycloalkenylgroups can include bicyclic and tricyclic alkenyl groups.
  • Alkenyl groups are optionally substituted.
  • Substituted alkenyl groups include among others those which are substituted with alkyl or aryl groups, which groups in turn can be optionally substituted.
  • alkenyl groups include ethenyl, prop-1-enyl, prop- 2-enyl, cycloprop-1 -enyl, but-1-enyl, but-2-enyl, cyclobut-1-enyl, cyclobut-2-enyl, pent-1-enyl, pent-2-enyl, branched pentenyl, cyclopent-1 -enyl, hex-1-enyl, branched hexenyl, cyclohexenyl, all of which are optionally substituted.
  • Substituted alkenyl groups include fully halogenated or semihalogenated alkenyl groups, such as alkenyl groups having one or more hydrogens replaced with one or more fluorine atoms, chlorine atoms, bromine atoms and/or iodine atoms.
  • Substituted alkenyl groups include fully fluorinated or semifluorinated alkenyl groups, such as alkenyl groups having one or more hydrogen atoms replaced with one or more fluorine atoms.
  • Aryl groups include groups having one or more 5-, 6- or 7- member aromatic and/or heterocyclic aromatic rings.
  • heteroaryl specifically refers to aryl groups having at least one 5-, 6- or 7- member heterocyclic aromatic rings.
  • Aryl groups can contain one or more fused aromatic and heteroaromatic rings or a combination of one or more aromatic or heteroaromatic rings and one or more nonaromatic rings that may be fused or linked via covalent bonds.
  • Heterocyclic aromatic rings can include one or more N, O, or S atoms in the ring.
  • Heterocyclic aromatic rings can include those with one, two or three N atoms, those with one or two O atoms, and those with one or two S atoms, or combinations of one or two or three N, O or S atoms.
  • Aryl groups are optionally substituted.
  • Substituted aryl groups include among others those which are substituted with alkyl or alkenyl groups, which groups in turn can be optionally substituted.
  • aryl groups include phenyl, biphenyl groups, pyrrolidinyl, imidazolidinyl, tetrahydrofuryl, tetrahydrothienyl, furyl, thienyl, pyridyl, quinolyl, isoquinolyl, pyridazinyl, pyrazinyl, indolyl, imidazolyl, oxazolyl, thiazolyl, pyrazolyl, pyridinyl, benzoxadiazolyl, benzothiadiazolyl, and naphthyl groups, all of which are optionally substituted.
  • Substituted aryl groups include fully halogenated or semihalogenated aryl groups, such as aryl groups having one or more hydrogens replaced with one or more fluorine atoms, chlorine atoms, bromine atoms and/or iodine atoms.
  • Substituted aryl groups include fully fluorinated or semifluorinated aryl groups, such as aryl groups having one or more hydrogens replaced with one or more fluorine atoms.
  • Aryl groups include, but are not limited to, aromatic group-containing or heterocylic aromatic group-containing groups corresponding to any one of the following: benzene, naphthalene, naphthoquinone, diphenylmethane, fluorene, anthracene, anthraquinone, phenanthrene, tetracene, tetracenedione, pyridine, quinoline, isoquinoline, indoles, isoindole, pyrrole, imidazole, oxazole, thiazole, pyrazole, pyrazine, pyrimidine, purine, benzimidazole, furans, benzofuran, dibenzofuran, carbazole, acridine, acridone, phenanthridine, thiophene, benzothiophene, dibenzothiophene, xanthene, xanthone, flavone, coumarin, a
  • a group corresponding to the groups listed above expressly includes an aromatic or heterocyclic aromatic group, including monovalent, divalent and polyvalent groups, of the aromatic and heterocyclic aromatic groups listed herein are provided in a covalently bonded configuration in the compounds of the invention at any suitable point of attachment.
  • aryl groups contain between 5 and 30 carbon atoms.
  • aryl groups contain one aromatic or heteroaromatic six-membered ring and one or more additional five- or six-membered aromatic or heteroaromatic ring.
  • aryl groups contain between five and eighteen carbon atoms in the rings.
  • Aryl groups optionally have one or more aromatic rings or heterocyclic aromatic rings having one or more electron donating groups, electron withdrawing groups and/or targeting ligands provided as substituents.
  • Arylalkyl groups are alkyl groups substituted with one or more aryl groups wherein the alkyl groups optionally carry additional substituents and the aryl groups are optionally substituted.
  • Specific alkylaryl groups are phenyl-substituted alkyl groups, e.g., phenylmethyl groups.
  • Alkylaryl groups are alternatively described as aryl groups substituted with one or more alkyl groups wherein the alkyl groups optionally carry additional substituents and the aryl groups are optionally substituted.
  • Specific alkylaryl groups are alkyl-substituted phenyl groups such as methylphenyl.
  • Substituted arylalkyl groups include fully halogenated or semihalogenated arylalkyl groups, such as arylalkyl groups having one or more alkyl and/or aryl groups having one or more hydrogens replaced with one or more fluorine atoms, chlorine atoms, bromine atoms and/or iodine atoms.
  • any of the groups described herein which contain one or more substituents do not contain any substitution or substitution patterns which are sterically impractical and/or synthetically non-feasible.
  • the compounds of this invention include all stereochemical isomers arising from the substitution of these compounds.
  • Optional substitution of alkyl groups includes substitution with one or more alkenyl groups, aryl groups or both, wherein the alkenyl groups or aryl groups are optionally substituted.
  • Optional substitution of alkenyl groups includes substitution with one or more alkyl groups, aryl groups, or both, wherein the alkyl groups or aryl groups are optionally substituted.
  • Optional substitution of aryl groups includes substitution of the aryl ring with one or more alkyl groups, alkenyl groups, or both, wherein the alkyl groups or alkenyl groups are optionally substituted.
  • Optional substituents for any alkyl, alkenyl and aryl group includes substitution with one or more of the following substituents, among others:
  • halogen including fluorine, chlorine, bromine or iodine
  • R is a hydrogen or an alkyl group or an aryl group and more specifically where R is a methyl, ethyl, propyl, butyl, or phenyl group all of which groups are optionally substituted;
  • R is a hydrogen or an alkyl group or an aryl group and more specifically where R is a methyl, ethyl, propyl, butyl, or phenyl group all of which groups are optionally substituted;
  • each R independently of each other R, is a hydrogen or an alkyl group or an aryl group and more specifically where R is a methyl, ethyl, propyl, butyl, or phenyl group all of which groups are optionally substituted; and where R and R can form a ring which can contain one or more double bonds and can contain one or more additional carbon atoms;
  • each R independently of each other R, is a hydrogen or an alkyl group or an aryl group and more specifically where R is a methyl, ethyl, propyl, butyl, or phenyl group all of which groups are optionally substituted; and where R and R can form a ring which can contain one or more double bonds and can contain one or more additional carbon atoms;
  • each R independently of each other R, is a hydrogen, or an alkyl group, or an acyl group or an aryl group and more specifically where R is a methyl, ethyl, propyl, butyl, phenyl or acetyl group, all of which are optionally substituted; and where R and R can form a ring which can contain one or more double bonds and can contain one or more additional carbon atoms;
  • -SR where R is hydrogen or an alkyl group or an aryl group and more specifically where R is hydrogen, methyl, ethyl, propyl, butyl, or a phenyl group, which are optionally substituted;
  • R is an alkyl group or an aryl group and more specifically where R is a methyl, ethyl, propyl, butyl, or phenyl group, all of which are optionally substituted;
  • each R independently of each other R, is a hydrogen, or an alkyl group, or an aryl group all of which are optionally substituted and wherein R and R can form a ring which can contain one or more double bonds and can contain one or more additional carbon atoms;
  • R is H, an alkyl group, an aryl group, or an acyl group all of which are optionally substituted.
  • R can be an acyl yielding -OCOR" where R" is a hydrogen or an alkyl group or an aryl group and more specifically where R" is methyl, ethyl, propyl, butyl, or phenyl groups all of which groups are optionally substituted.
  • Specific substituted alkyl groups include haloalkyl groups, particularly trihalomethyl groups and specifically trifluoromethyl groups.
  • Specific substituted aryl groups include mono-, di- , tri, tetra- and pentahalo-substituted phenyl groups; mono-, di-, tri-, tetra-, penta-, hexa-, and hepta-halo-substituted naphthalene groups; 3- or 4-halo-substituted phenyl groups, 3- or 4-alkyl- substituted phenyl groups, 3- or 4-alkoxy-substituted phenyl groups, 3- or 4-RCO-substituted phenyl, 5- or 6-halo-substituted naphthalene groups.
  • substituted aryl groups include acetylphenyl groups, particularly 4-acetylphenyl groups; fluorophenyl groups, particularly
  • inflammation generally refers to a biological response of tissues to harmful stimuli, such as pathogens, damaged cells, irritants, etc. Inflammation can be either acute or chronic. Acute inflammation is an initial response of the body to harmful stimuli and can be achieved by the increased movement of plasma and leukocytes from the blood into injured tissues. An inflammatory response can involve the local vascular system, the immune system, and/or various cells within the injured tissue. Prolonged inflammation, referred to as chronic inflammation, can lead to a progressive shift in the type of cells which are present at the site of inflammation can be characterized by simultaneous destruction and healing of the tissue from the inflammatory process.
  • amino acid comprises naturally occurring amino acids as well as non- naturally occurring amino acids, including amino acid analogs and derivatives.
  • amino acid analogs and derivatives include, for example, naturally occurring proteogenic L-amino acids; D-amino acids; chemically modified amino acids such as amino acid analogs and derivatives; naturally occurring non-proteogenic amino acids, and chemically synthesized compounds having properties known in the art to be characteristic of amino acids.
  • nucleic acid generally refers to a molecule or strand of DNA, RNA, or derivatives or analogs thereof including one or more nucleobases. Nucleobases comprise purine or pyrimidine bases typically found in DNA or RNA (e.g., adenine, guanine, thymine, cytosine, and/or uracil). The term “nucleic acid” also comprises oligonucleotides and polynucleotides. Nucleic acids may be single-stranded molecules, or they may be double-, triple- or quadruple-stranded molecules that may comprise one or more complementary strands of a particular molecule.
  • Nucleic acid includes artificial nucleic acids including peptide nucleic acids, morpholino nucleic acids, glycol nucleic acids and threose nucleic acids. Artificial nucleic acids may be capable of nucleic acid hybridization.
  • sequence means the linear order in which monomers occur in a polymer, the order of amino acids in a polypeptide or the order of nucleotides in a polynucleotide for example.
  • peptide and “polypeptide” are used synonymously in the present description, and refer to a class of compounds comprising of amino acid residues chemically bonded together by amide bonds (or peptide bonds), regardless of length, functionality, environment, or associated molecule(s).
  • Peptides and polypeptides are polymeric compounds comprising at least two amino acid residues or modified amino acid residues. Modifications can be naturally occurring or non-naturally occurring, such as modifications generated by chemical synthesis.
  • Modifications to amino acids in peptides include, but are not limited to, phosphorylation, glycosylation, lipidation, prenylation, sulfonation, hydroxylation, acetylation, methionine oxidation, alkylation, acylation, carbamylation, iodination and the addition of cofactors.
  • Peptides include proteins and further include compositions generated by degradation of proteins, for example by proteolyic digestion. Peptides and polypeptides can be generated by substantially complete digestion or by partial digestion of proteins.
  • Polypeptides comprising 2 to 100 amino acid units, optionally for some embodiments 2 to 50 amino acid units and, optionally for some embodiments 2 to 20 amino acid units can be used as polypeptide targeting ligands in the invention, for example, where the polypepetide preferentially binds to proteins, peptides or other biomolecules expressed, or otherwise generated by, a target tissue, such as a tumor, precancerous tissue, site of inflammation or other lesion.
  • the polypeptide is at least four amino acid residues in length and can range up to a full-length protein.
  • Protein refers to a class of compounds comprising one or more polypeptide chains and/or modified polypeptide chains. Proteins can be modified by naturally occurring processes such as post-translational modifications or co-translational modifications. Exemplary post- translational modifications or co-translational modifications include, but are not limited to, phosphorylation, glycosylation, lipidation, prenylation, sulfonation, hydroxylation, acetylation, methionine oxidation, the addition of cofactors, proteolysis, and assembly of proteins into macromolecular complexes. Modification of proteins can also include non-naturally occurring derivatives, analogues and functional mimetics generated by chemical synthesis. Exemplary derivatives include chemical modifications such as alkylation, acylation, carbamylation, iodination or any modification that derivatizes the protein.
  • polynucleotide and “oligonucleotide” are used interchangeably and refer to a class of compounds composed of nucleic acid residues chemically bonded together.
  • the invention provides optical agents having an oligonucleotide or polynucleotide targeting ligand which comprises a plurality of nucleic acid residues, such as DNA or RNA residues, and/or modified nucleic acid residues that preferentially binds to proteins, peptides or other biomolecules expressed, or otherwise generated by, a target tissue, such as a tumor, precancerous tissue, site of inflammation or other lesion.
  • Oligo- or poly-nucleotide targeting ligands include, for example, oligo- or poly-nucleotides comprising 2 to 100 nucleic acid units, optionally for some embodiments 2 to 50 nucleic acid units and, optionally for some embodiments 2 to 20 nucleic acid units, and optionally for some embodiments 2 to 10 nucleic acid units.
  • Polypeptide and oligonucleotide include a polymer of at least two nucleotides joined together by phosphodiester bonds and may consist of either ribonucleotides or deoxyribonucleotides.
  • aptamer refers to an oligo- or poly-nucleotide or polypeptide that binds to, or otherwise selectively or preferentially associates with, a specific target molecule.
  • the invention provides optical agents having an aptamer targeting ligand that preferentially binds to proteins, peptides or other biomolecules expressed, or otherwise generated by, a target tissue, such as a tumor, precancerous tissue, site of inflammation or other lesion.
  • Peptidomimetic refers to a molecule having activity, including biological activity, that resembles that of a polypeptide or is substantially the same as a polypeptide.
  • Morphine for example, is a peptidomimetic of endorphin peptide.
  • a peptidomimetic is a small protein-like polymer designed to mimic the functionality of a peptide.
  • Peptidomimetics useful as targeting ligands for some compounds of the invention in the present invention include peptoids and ⁇ -peptides. The composition and biological activity of peptidomimetics and use of peptidomimetics in targeted diagnostics and therapeutics are further described in the following references: (1 ) A. Giannis and T.
  • attaching moiety refers to a component provided to attach an optical dye, such as a pyrazine, cyanine, azulene, azaazulene or indocyanine group, directly or indirectly to central sulfone hydroxamic acid or hydroxamate backbone in compounds of the invention.
  • an optical dye such as a pyrazine, cyanine, azulene, azaazulene or indocyanine group
  • L 1 , L 2 , W 1 and/or W 2 in formulas (FX1 ) - (FX113) are an attaching moieties.
  • an “electron withdrawing group” refers to a chemical group that draws electrons or electron density from a center, such as optical dye component of compounds of the invention, for example a pyrazine, cyanine, azulene, azaazulene or indocyanine group.
  • the electron withdrawing group(s) are independently selected from cyano (-CN), carbonyl (-CO), carboxylate (-C0 2 R a ), halo (-F, -CI, -Br, -I, -At), carbamate (-CONR b R c ), acyl (-COR d ), nitro (-N0 2 ), sulfinyl (-SOR e ), sulfonyl (- S0 2 R , -SO2OR 9 , and -POsR ⁇ , wherein in the context of this description, R a to R' are independently selected to enhance biological and/or physiochemical properties of the optical agents of the invention.
  • R a to R' are independently selected from any one of a hydrogen atom, an anionic functional group (e.g., carboxylate, sulfonate, sulfate, phosphonate or phosphate) and a hydrophilic functional group (e.g., hydroxyl, carboxyl, sulfonyl, sulfonato or phosphonato).
  • an anionic functional group e.g., carboxylate, sulfonate, sulfate, phosphonate or phosphate
  • a hydrophilic functional group e.g., hydroxyl, carboxyl, sulfonyl, sulfonato or phosphonato.
  • the EWG(s) are independently selected from is -CN, -C0 2 R 40 , - S0 2 OR 43 , -CONR 52 R 53 , -COR 54 , -N0 2 , -SOR 41 ; -S0 2 R 55 , -POsR ⁇ R 45 , halo, C C 6 acyl, trihalomethyl, or -S0 2 NR 56 R 57 , wherein each of R 40 to R 59 is independently H or d - C 10 alkyl.
  • an EWG is located at the terminus of a substituent arm of a an optical dye of compounds of formulas (FX1 ) - (FX113), such pyrazine, cyanine, azulene, azaazulene or indocyanine group of the compounds of compounds of formulas (FX1 ) - (FX113).
  • an “electron donating group” refers to a chemical group that releases electrons or electron density to a center, such as optical dye component of compounds of the invention, for example a pyrazine, cyanine, azulene, azaazulene or indocyanine group.
  • the electron donating group(s) are independently selected from C1-C10 alkyl, C5-C10 aryl, -(CH 2 ) z OH, -OR j , -SR k , -NR'R m , - N(R n )COR°, and -P(R ), wherein in the context of this description, R j to R are independently selected to enhance biological and/or physiochemical properties of the optical agents of the invention and wherein z is selected from the range of 1 to 10.
  • R j to R are independently selected from any one of a hydrogen atom, an anionic functional group (e.g., carboxylate, sulfonate, sulfate, phosphonate or phosphate) and a hydrophilic functional group (e.g., hydroxyl, carboxyl, sulfonyl, sulfonato or phosphonato).
  • an anionic functional group e.g., carboxylate, sulfonate, sulfate, phosphonate or phosphate
  • a hydrophilic functional group e.g., hydroxyl, carboxyl, sulfonyl, sulfonato or phosphonato.
  • the EDG(s) are independently C C 6 alkyl, C 3 -C 6 cycloalkyl -OR 46 , -SR 47 , -NR 48 R 49 , or -NR 50 COR 51 , wherein each of R 46 to R 51 is independently H or C-i - C 10 alkyl.
  • an EDG is located at the terminus of a substituent arm of a substituent arm of a an optical dye of compounds of formulas (FX1 ) - (FX1 13), such pyrazine, cyanine, azulene, azaazulene or indocyanine group of the compounds of compounds of formulas (FX1 ) - (FX1 13).
  • two substituents, such as EDG and EWG substituents, on a compound of the invention can act in what is known as a "push-pull" arrangement.
  • the electron density of the compound or a portion thereof, such as an aryl or heteroaryl group is polarized due in part to the location of an EWG and EDG on the compound.
  • an EWG is positioned at a terminus of a substituent arm of the structure and an EDG is positioned at a terminus of a different substituent arm of the structure.
  • an EWG is positioned at one end of a ⁇ bond and an EDG is positioned at the other end of a ⁇ bond.
  • an EWG is positioned para- to an EDG in a six- membered ring structure.
  • an EWG is positioned trans- to an EDG in an alkylene structure.
  • compounds having the "push-pull" arrangement exhibit a shift in the optical absorbance and emission spectrum as compared to compounds not having the "push-pull" arrangement.
  • optical agent generally refers to compounds, compositions, preparations, and/or formulations that absorb, emit, or scatter electromagnetic radiation of wavelength generally in the range of 350 - 1300 nanometers, within a biologically relevant environment or condition.
  • optical agents of the invention when excited by electromagnetic radiation, undergo emission via fluorescence or phosphorescence pathways. These pathways are useful for diagnostic imaging, visualization, or organ function monitoring. Compounds belonging to this class are commonly referred to as “optical imaging agents” or “optical contrast agents.”
  • optical agents of the invention absorb electromagnetic radiation and undergo photochemical reactions such as photofragmentation of one or more photolabile bonds to generate reactive species such as nitrenes, carbene, free radicals, ions, excited species, etc.
  • photosensitizers refers to a phototherapeutic agent or a component thereof providing for photoactivation, for example, photoactivation resulting in generation of reactive species that locally kill, injure, inactivate or otherwise degrade cells (e.g., cancer cells, tumor cells, non-cancer cells, etc.).
  • Photosensitizers of some embodiments undergo photoactivation that initiates bond cleavage reactions, such as photolysis and/or nitrogen extrusion reactions, thereby generating reactive species capable of causing localized cell death or injury.
  • Optical agents include Type 1 and Type 2 phototherapeutic agents.
  • Optical agents include, but are not limited to, phototherapeutic agents (Type 1 and 2), photosensitizers, imaging agents, dyes, detectable agents, photosensitizer agents, photoactivators, and photoreactive agents; and conjugates, complexes, and derivatives thereof.
  • chromophore is a compound or functional group of a compound that results in absorption of electromagnetic radiation, preferably for some applications electromagnetic radiation having wavelengths in the UV (e.g. 200 nm to 350 nm) or visible (e.g. 350 nm to 750 nm) of the electromagnetic spectrum.
  • a fluorophore is a compound or functional group of a compound that results in absorption of electromagnetic radiation and subsequent fluorescence.
  • incorporation of a fluorophore results in compounds of the invention that absorb electromagnetic radiation and generate fluorescence having wavelengths in the UV (e.g. 200 nm to 350 nm) or visible (e.g. 350 nm to 750 nm) of the electromagnetic spectrum.
  • incorporation of a fluorophore results in compounds having an appreciable quantum yield for fluorescence, such as a quantum yield over the range of 0.001 to 1 , 0.01 to 1 , optionally 0.1 to 1.
  • Optical agents of the present invention can contain fluorophores.
  • Fluorophores can be functional groups in a molecule which absorb electromagnetic radiation of first specific wavelengths and re-emit energy at second specific wavelengths. The amount and wavelengths of the emitted electromagnetic radiation depend on both the fluorophore and the chemical environment of the fluorophore.
  • fluorophore may be abbreviated throughout the present description as "FL".
  • fluorophores emit energy in the visible (e.g. 350 nm to 750 nm) and NIR regions (e.g., 750 - 1300nm) of the electromagnetic spectrum.
  • Luminescence refers to the emission of electromagnetic radiation from excited electronic states of atoms or molecules.
  • Luminescence generally refers to electromagnetic radiation emission, such as photoluminescence, chemiluminescence, and electrochemiluminescence, among others.
  • photoluminescence including fluorescence and phosphorescence
  • the excited electronic state is created by the absorption of electromagnetic radiation.
  • Luminescence detection involves detection of one or more properties of the luminescence or associated luminescence process. These properties can include intensity, excitation and/or emission spectrum, polarization, lifetime, and energy transfer, among others. These properties can also include time-independent (steady-state) and/or time-dependent (time- resolved) properties of the luminescence.
  • Representative luminescence techniques include fluorescence intensity (FLINT), fluorescence polarization (FP), fluorescence resonance energy transfer (FRET), fluorescence lifetime (FLT), total internal reflection fluorescence (TIRF), fluorescence correlation spectroscopy (FCS), fluorescence recovery after photobleaching (FRAP), and bioluminescence resonance energy transfer (BRET), among others.
  • FLINT fluorescence intensity
  • FP fluorescence polarization
  • FRET fluorescence resonance energy transfer
  • FLT fluorescence lifetime
  • TIRF total internal reflection fluorescence
  • FCS fluorescence correlation spectroscopy
  • FRAP fluorescence recovery after photobleaching
  • BRET bioluminescence resonance energy transfer
  • Such emission can be detected using fluorometric techniques as described above.
  • One skilled in the art can readily determine the most appropriate treatment and optional detection technique based, at least in part, on the specific phototherapeutic agent(s) administered and/or the particular use (e.g., tissue to be treated).
  • Optical condition refers to one or more of the following: the fluorescence quantum yield, fluorescence intensity, fluorescence excitation wavelength, wavelength distribution or spectrum, emission wavelength, wavelength distribution or spectrum, Stokes shift, color, reflectance, phosphorescence, chemiluminescence, scattering, and/or other observable and/or measurable spectral property or phenomenon.
  • Phototherapy procedure refers to a therapeutic procedure involving administration of a phototherapeutic agent to a patient followed by subsequent excitation by exposure to applied electromagnetic radiation, such as electromagnetic radiation having wavelengths in the visible and/or near IR region of the electromagnetic spectrum. Such wavelengths can be in the range of 350 - 1300 nanometers, so as to generate a therapeutically effective amount of excited phototherapeutic agent.
  • Phototherapy includes, but is not limited to, photodynamic therapy.
  • phototherapy includes procedures involving administration of Type 1 and/or Type 2 phototherapeutic agents, optionally further including administration of one or more additional therapeutic agents.
  • a detectable optical signal may be, for example, an observable change in absorbance, reflectance, phosphorescence, chemiluminescence, scattering, or other spectral property.
  • tumor-specific agent refers to a compound or composition, such as an optical agent, that preferentially accumulates in a tumor at a higher level than normal tissue regardless of the particular mechanism of uptake in the tumors, for example, receptor mediated or enhanced permeability and retention (EPR).
  • Optical agents of the invention include tumor- specific agents, including tumor specific phototherapy agents, for example having a targeting ligand providing specificity in the administration, delivery and/or binding to tumor tissue.
  • targeting ligand refers to a chemical group and/or substituent having functionality for targeting a compound of the invention to an anatomical and/or physiological site of a patient, such as a selected cell, tissue or organ.
  • a targeting ligand is characterized as a ligand that selectively or preferentially binds to a specific biological site(s) (e.g., enzymes, receptors, etc.) and/or biological surface(s) (e.g., membranes, fibrous networks, etc.).
  • Target tissue refers to tissue of a subject to which an optical agent is administered or otherwise contacted, for example during a biomedical procedure such as an optical imaging, phototherapy, monitoring or visualization procedure.
  • Target tissues can be contacted with an optical agent of the invention under in vivo conditions in vitro conditions or ex vivo conditions.
  • Target tissues in some embodiments include cancerous tissue, cancer cells, precancerous tissue, a tumor, a lesion, a site of inflammation, or vasculature tissue.
  • a target tissue includes a melanoma cell, a breast lesion, a prostate lesion, a lung cancer cell, a colorectal cancer cell, an atherosclerotic plaque, a brain lesion, a blood vessel lesion, a lung lesion, a heart lesion, a throat lesion, an ear lesion, a rectal lesion, a bladder lesion, a stomach lesion, an intestinal lesion, an esophagus lesion, a liver lesion, a pancreatic lesion, and a solid tumor.
  • Target tissue in some embodiments refers to a selected organ of the subject or component thereof, such as lung, heart, brain, stomach, liver, kidneys, gallbladder, pancreas, intestines, rectum, skin, colon, prostate, ovaries, breast, bladder, blood vessel, throat, ear, or esophagus.
  • Methods of this invention comprise the step of administering an "effective amount" of the present diagnostic and therapeutic compositions, formulations and preparations containing the present compounds or compositions, to diagnose, image, monitor, evaluate, treat, reduce, alleviate, ameliorate or regulate a biological condition and/or disease state in a patient.
  • the term "effective amount,” as used herein, refers to the amount of the diagnostic and therapeutic formulation, that, when administered to the individual is effective to diagnose, image, monitor, evaluate, treat, reduce alleviate, ameliorate or regulate a biological condition and/or disease state.
  • an effective amount of a given composition or formulation will depend at least in part upon the mode of administration (e.g.
  • any carrier or vehicle employed any carrier or vehicle employed, and the specific individual to whom the formulation is to be administered (age, weight, condition, sex, etc.).
  • the dosage requirements needed to achieve the "effective amount" vary with the particular formulations employed, the route of administration, and clinical objectives. Based on the results obtained in standard pharmacological test procedures, projected daily dosages of active compound or composition can be determined as is understood in the art.
  • an effective amount of a compound or composition of the invention is a therapeutically effective amount.
  • the phrase "therapeutically effective” qualifies the amount of compound or composition administered in the therapy. This amount achieves the goal of ameliorating, suppressing, eradicating, preventing, reducing the risk of, or delaying the onset of a targeted condition.
  • an effective amount of a compound or composition of the invention is a diagnostically effective amount.
  • the phrase "diagnostically effective" qualifies the amount of compound or composition administered in diagnosis, for example of a disease state or other pathological condition. The amount achieves the goal of being detectable while avoiding adverse side effects found with higher doses.
  • an active ingredient or other component is included in a therapeutically acceptable amount.
  • an active ingredient or other component is included in a diagnostically acceptable amount.
  • the compounds and pharmaceutically acceptable salts of the invention can be used as part of a combination.
  • the term "combination" means the administration of two or more compounds directed to a target condition.
  • the treatments of the combination generally can be co-administered in a simultaneous manner.
  • Two compounds can be co-administered as, for example: (a) a single formulation (e.g., a single capsule) having a fixed ratio of active ingredients; or (b) multiple, separate formulations (e.g., multiple capsules) for each compound.
  • the treatments of the combination can alternatively (or additionally) be administered at different times.
  • the invention encompasses administering optical agents useful in the invention to a patient or subject.
  • a "patient” or “subject”, used equivalently herein, refers to an animal.
  • an animal refers to a mammal, preferably a human.
  • the subject can either: (1 ) have a condition able to be monitored, diagnosed, prevented and/or treated by administration of an optical agent of the invention; or (2) is susceptible to a condition that is able to be monitored, diagnosed, prevented and/or treated by administering an optical agent of the invention.
  • diagnosis When used herein, the terms "diagnosis”, “diagnostic” and other root word derivatives are as understood in the art and are further intended to include a general monitoring, characterizing and/or identifying a state of health or disease. The term is meant to encompass the concept of prognosis.
  • diagnosis of cancer can include an initial determination and/or one or more subsequent assessments regardless of the outcome of a previous finding. The term does not necessarily imply a defined level of certainty regarding the prediction of a particular status or outcome.
  • administering means that a compound or formulation thereof of the invention, such as an optical agent, is provided to a patient or subject, for example in a therapeutically effective amount.
  • the invention includes methods for a biomedical procedure wherein a therapeutically or diagnostically effective amount of a compound having any one of formulas (FX1 ) - (FX113) is administered to a patient in need of treatment, for example to a patient undergoing treatment for a diagnosed diseased state including cancer and vascular diseases.
  • Administering can be carried out by a range of techniques known in the art including parenteral administration including intravenous, intraperitoneal or subcutaneous injection or infusion, oral administration, topical or transdermal absorption through the skin, or by inhalation, for example.
  • parenteral administration including intravenous, intraperitoneal or subcutaneous injection or infusion, oral administration, topical or transdermal absorption through the skin, or by inhalation, for example.
  • the chosen route of administration may depend on such factors as solubility of the compound or composition, location of targeted condition, and other factors which are within the knowledge of one having ordinary skill in the relevant art.
  • Topical administration includes the use of transdermal administration, such as transdermal patches or iontophoresis devices.
  • Parenteral administration includes subcutaneous injections, intravenous injections, intraarterial injections, intraorbital injections, intracapsular injections, intraspinal injections, intraperitoneal injections, intramuscular injections, intrasternal injections, and infusion.
  • Dosage forms suitable for parenteral administration include solutions, suspensions, dispersions, emulsions, and any other dosage form that can be administered parenterally.
  • controlled-release component refers to an agent that facilitates the controlled-release of a compound including, but not limited to, polymers, polymer matrices, gels, permeable membranes, liposomes, microspheres, or the like, or any combination thereof. Methods for producing compounds in combination with controlled-release components are known to those of skill in the art.
  • the term "pharmaceutically acceptable” means approved by a regulatory agency of an appropriate federal or state government; or listed in the U.S. Pharmacopoeia or other generally recognized pharmacopoeia for use in animals, and more particularly in humans; or does not impart significant deleterious or undesirable effect on a subject to whom it is administered and in the context in which it is administered.
  • the groups and structures described herein as portions of the compounds of the invention may be defined as if they are separate valence-satisfied chemical structures. It is intended that when a group is described or shown as being a substituent of another group, that the group be viewed as having a valency to allow this binding to occur.
  • Example 1 Optically Functional Matrix Metalloproteinase Inhibitors
  • MMPs Matrix metalloproteinases
  • MMPs are zinc-dependent enzymes responsible for the remodeling and degradation of components of the extracellular matrix.
  • the up-regulation of MMPs has been implicated in numerous diseases including osteoarthritis and cancer.
  • a key therapeutic target for MMPs is cancer, as MMPs appear important for tumor growth and metastasis processes, and inhibition of MMP-9 in particular appears able to block metastasis.
  • clinical trials with MMP inhibitors for advanced cancer(s) have not been successful in demonstrating efficacy.
  • Bramhall has now reported a placebo-controlled double blind study reporting success in treating cancer with an MMP inhibitor in a study treating gastric cancer patients with the broad spectrum inhibitor marimastat.
  • Marimastat afforded a survival rate similar to Gemcitabine in patients with unresectable pancreatic cancer. [Bramhall, S.R. et al., Marimastat as first-line therapy for patients with unresectable pancreatic cancer: a randomized trial, J. Clin. Oncol., 2001 , 19, 3447-3455]. Thus, the proof-of principle for efficacy in treating human cancers with MMP inhibitors appears to have been clinically demonstrated.
  • MMPs have been shown to be up-regulated; most notably in osteoarthritis and in cardiovascular disease as there is significant up-regulation of MMP-2 and MMP-13 immediately after an ischemic event. There is also evidence that treatment with an MMP inhibitor immediately after an ischemic event for a defined acute period has significant beneficial effects on left ventricular re-modeling. This therapeutic approach may have potential in an intensive care unit environment.
  • MMP-2 and MMP-9 have recently been implicated in neuropathic pain states; both acute and chronic. A way to image the neuropathic pain pathway via imaging tissue expressing MMPs would also be beneficial as there are many aspects of the fundamental biology that are poorly understood
  • Compounds of the present invention include optical agents providing target-specific small molecule optical imaging agents that inhibit the biological activity of various MMPs.
  • the present compounds are useful for optical evaluation and monitoring of biological materials where MMPs play a role in biological pathways related to the above mentioned diseases and conditions.
  • the invention includes methods of using optically functional MMP inhibitors to improve patient outcomes by enabling imaging and visualization techniques, optionally in combination with other anticancer or cardiovascular (or other disease where coadministration of above mentioned MMP Inhibitor would be beneficial) drug treatment regimens.
  • Optical agents of some embodiments comprise a sulfone hydroxamic acid or hydroxamate backbone and a fluorescent moiety having substituents providing for optical functionality useful for optical imaging, therapy and/or detection.
  • the sulfone hydroxamic acid or hydroxamate backbone and a fluorescent moiety are covalently linked such that the molecular recognition functionality for MMPs is preserved.
  • an optical dye comprising a fluorescent moiety is provided in the solvent exposed portion of the MMP binding pocket and in other embodiments an optical dye comprising a fluorescent moiety is provided in the S1 ' MMP binding pocket.
  • optically functional matrix metalloproteinase inhibitors useful for detection, imaging and diagnosis of target tissue expressing MMPs are provided in Table 1 .
  • R 3 , R 6 , R 16 , R ; and a are as set forth and described in the context of formulas (FX1 ) - (FX113).
  • TNF-a Converting Enzyme TNF-a Converting Enzyme
  • MMP-2 and MMP-9 To evaluate their inhibitory activity for MMPs, a number of optically functional MMP inhibitors were synthesized and characterized with respect to their percent inhibition of specific binding or activity maximal inhibitory concentration (IC 50 ) with respect to MMP-2 and MMP-9. Compounds evaluated were also characterized with respect to certain optical properties including maximum absorption wavelength ( ⁇ ) ) and maximum emission wavelength ( ⁇ ( ⁇ ) ) in visible and infrared regions.
  • Biochemical assay results are presented as the percent inhibition of specific binding or activity. All other results are expressed in terms of that assay's quantitation method. For primary assays, only the lowest concentration with a significant response judged by the assays' criteria, is shown. Where applicable, either the secondary assay results with the lowest
  • Table 5 summarizes percentage inhibition values obtained for compounds having formula (FX105) - (FX109).
  • Figures 1 - 5 provide response curves obtained for some of the compounds evaluated. Closed circles in Figures 1-5 represent percentage inhibition values for compounds of the present invention. The closed squares in Figures 1-5 represent percentage inhibition values for control measurements for TIMP-2 (Tissue Inhibitor Metalloprotease-2) which is a natural regulator of MMPs.
  • Figure 1 provides the response curve for inhibition of MMP-2 by compound (FX107).
  • Figure 2 provides the response curve for inhibition of MMP-9 by compound (FX107).
  • Figure 3 provides the response curve for inhibition of MMP-2 by compound (FX108).
  • Figure 4 provides the response curve for inhibition of MMP-2 by compound (FX105).
  • Figure 5 provides the response curve for inhibition of MMP-9 by compound (FX105).
  • percentage inhibition (%) is plotted as a function of concentration ( ⁇ ) of the optically functional MMP inhibitor compound.
  • Table 6 summarizes experimental results in connection with characterization of the inhibitor activity of the compounds evaluated. Parameters provided in Table 6 include percentage inhibition and half maximal inhibitory concentration (IC 50 ) in connection with selective inhibition of MMP-2 and MMP-9.
  • the optically functional MMP inhibitors exhibit good activity (e.g., IC 50 ⁇ 10 nM) with respect to MMP-2 and MMP-9. It is important to note that comparable activity was observed for MMP inhibitors having pyrazine and indocyanine groups, suggesting that the presence of the optical dye component does not appreciable impact selective binding. In addition, comparable activity was observed for MMP inhibitors with and without a short spacer positioned between the hydroxamic acid backbone and the optical dye components.
  • the present invention provides methods of using optically functional MMP inhibitors that selectively bind to MMP enzymes.
  • Such optically functional MMP inhibitors can be used for detecting, monitoring and imaging target tissue and cells, and in particular cancer cells, that express MMPs at higher levels than normal cells.
  • these optical agents provide MMP inhibitors useful for treatment of pathological conditions associated with MMP enzymes, such as cancer and inflammation.
  • the sulfone hydroxamic acid or hydroxamate backbone of the optically functional MMP inhibitors serve to bind selectively the MMPs, whereas the fluorescent moiety functions as an optical tag or probe that enables detection or imaging of tissue and cells that express MMPs.
  • molecules absorbing, emitting, or scattering in the visible, NIR, or long-wavelength (UV-A, > 300 nm) region of the electromagnetic spectrum are useful for optical detection and/or imaging.
  • the high sensitivity associated with fluorescence permits detection without the negative effects of radioactivity or ionizing radiation.
  • Pyrazines, cyanines, azulenes, azaazulenes and indocyanines are one of the few classes of small molecules having desirable photophysical properties for biomedical optical applications.
  • Binding of the dye component containing MMP inhibitors to MMPs may enhance fluorescence yield by immobilizing the dye component thereby minimizing non-radiative energy loss pathways.
  • the invention provides a method of using an optical agent, for example, in a biomedical procedure for optically imaging or visualizing a target tissue or a class of target tissues.
  • the present methods include tissue selective imaging and visualization methods, such as imaging or visualization of a target tissue that expresses, secretes or otherwise produces MMPs, for example, a target tissue that expresses, secretes or otherwise produces the MMPs at an elevated level relative to production corresponding to a health (i.e. non-disease) condition.
  • a method of this aspect comprises the step of administering a diagnostically effective amount of a compound to a subject, wherein the compound is a compound having any of formulas (FX1 ) - (FX113) or a pharmaceutical preparation thereof.
  • the method of this aspect further comprises contacting a target tissue that expresses, secretes or otherwise produces MMPs with a diagnostically effective amount of a compound having any of formulas (FX1 ) - (FX113).
  • the method of this aspect further comprises contacting cancer cells, for example, cancer cells of a tumor, with a diagnostically effective amount of a compound having any of formulas (FX1 ) - (FX113).
  • the administered compound upon administration, is optionally allowed to accumulate in a target region of interest (e.g., target cell, target tissue, target tumor, and/or target organ).
  • a target region of interest e.g., target cell, target tissue, target tumor, and/or target organ.
  • the present methods are useful for imaging or visualizing colorectal cancer and other cancers, including prostate cancer, gastric cancer, esophageal cancer, uterine- endometrial cancer, pancreatic cancer, breast cancer, cervical cancer, head and neck cancer, hepatic cancer, skin cancer, gallbladder cancer, lung cancer and ovarian cancer.
  • the compound that has been administered to the subject is exposed in vivo to electromagnetic radiation, thereby generating electromagnetic radiation emitted or scattered by the administered compound at the site of the target tissue.
  • the electromagnetic radiation emitted or scattered by the administered compound is then detected either visually or using optical detection instrumentation as known in the art.
  • fluorescence is excited from the compound (e.g., due to absorption of electromagnetic radiation exposed to the administered compound), optionally via multiphoton excitation processes.
  • the method of this aspect further comprises: (i) exposing a compound, such as a compound having any one of formula (FX1 ) - (FX113), administered to the subject to electromagnetic radiation capable of exciting emission from the compound; and (ii) detecting and/or measuring the intensity of emission from the compound.
  • a compound such as a compound having any one of formula (FX1 ) - (FX113)
  • the methods of the present invention use fluorescence excitation via exposure to light having wavelengths selected over the range of 300-1300 nm.
  • the wavelength of the electromagnetic radiation corresponds to a peak in the absorption spectrum of the optically functional MMP inhibitor, for example is within 20 nanometers of a peak in the absorption spectrum of the administered compound.
  • the target site is exposed to electromagnetic radiation having sufficient fluence and/or power sufficient to excite the optically functional MMP inhibitor so as to generate measurable fluorescence from the administered compound. Exposure to electromagnetic radiation and activation of the administered compound may occur during or after administration of the optically functional MMP inhibitor and accumulation at the target tissue.
  • OCT optical coherence tomography
  • OCT methods use wavelengths of about 1280 nm.
  • Use of electromagnetic radiation having wavelengths selected over the range of 700 nanometers to 1300 nanometers may be useful for some in situ optical imaging methods of the present invention, including biomedical applications for imaging organs, tissue and/or tumors, anatomical visualization, optical guided surgery and endoscopic procedures.
  • Compounds of the present methods may function as contrast agents, optical probes and/or tracer elements.
  • the methods of the present invention include in vivo, in vitro and ex vivo imaging and visualization.
  • the present invention provides methods for a range of clinical procedures, including optical imaging methods and/or visualization guided surgery and/or endoscopic diagnostic, monitoring and therapeutic procedures.
  • the electromagnetic radiation depends on the size, depth, and the pathology of the target tissue (e.g. tumor or lesion), as is known to one skilled in the art.
  • the fluence of the electromagnetic radiation is preferably, but not always, kept below 200 mW/cm 2 to minimize undesirable thermal effects.
  • the intensity, power, and duration of the illumination, and the wavelength of the electromagnetic radiation may vary widely depending on the body location, the lesions site, the effect to be achieved, etc.
  • the power of the applied electromagnetic radiation is preferably selected over the range of 1 - 500 mW/cm 2 and optionally selected over the range of 1 - 200 mW/cm 2 .
  • the duration of the exposure to applied electromagnetic radiation is selected over the range of 1 seconds to 60 minutes.
  • the electromagnetic radiation is applied internally, for example via endoluminal and endoscopic techniques.
  • Another aspect of the invention is a method of treating cancer, inflammation or inflammation-associated disorders, the method comprising administering to a subject in need thereof a therapeutically-effective amount of a compound having any of formulas (FX1 ) - (FX113) or a pharmaceutical preparation thereof.
  • These disorders may include osteoarthritis and rheumatoid arthritis.
  • the method of this aspect further comprises contacting a target tissue that expresses, secretes or otherwise produces MMPs with a therapeutically effective amount of a compound having any of formulas (FX1 ) - (FX113).
  • the method of this aspect comprises contacting cancer cells, for example, cancer cells of a tumor, with a therapeutically effective amount of a compound having any of formulas (FX1 ) - (FX113).
  • the administered compound upon administration, is optionally allowed to accumulate in a target region of interest (e.g., target tissue, tumor, or organ).
  • a target region of interest e.g., target tissue, tumor, or organ.
  • the invention provides compounds and methods for attenuating tumor progression and metastasis in animal tissues, comprising contacting a target tissue, such as tumor cells or tissues, with a therapeutically effective amount of an MMP inhibitor, such as a compound having any of formulas (FX1 ) - (FX113).
  • a target tissue such as tumor cells or tissues
  • an MMP inhibitor such as a compound having any of formulas (FX1 ) - (FX113).
  • the invention provides a method of reducing the risk of recurrence of a cancer or tumor after medical intervention (such intervention to include but not be limited to surgery, e.g. pulmonary surgery, surgical and endoscopic procedures, e.g.
  • the invention contemplates, for example, a method of minimizing post-operative recurrence of cancer in a patient, comprising administering to a cancer patient an MMP inhibitor, such as a compound having any of formulas (FX1 ) - (FX113).
  • the cancer is prostate cancer, gastrointestinal cancer, colon cancer, colorectal cancer, pancreatic cancer, breast cancer, lung cancer, throat cancer, skin cancer, stomach cancer, pancreatic cancer, brain cancer, liver cancer, prostate cancer, kidney cancer, bladder cancer, bone cancer, brain cancer, eye cancer, gallbladder cancer, head and neck cancer, Hodgkin lymphoma, mouth cancer, ovarian cancer, testicular cancer, throat cancer, esophageal cancer, small intestine cancer, pharyngeal cancer, laryngeal cancer, urethral cancer, uterine cancer, parathyroid cancer, cervical cancer, penile cancer or vaginal cancer.
  • the compounds of the present invention may also be useful for the treatment of cancer in patients, as described above, either when used alone or in combination with one or more other anticancer agents, e.g., radiotherapy and/or other chemotherapeutic, including
  • VEGF inhibitors include, but are not limited to, VEGF inhibitors, inhibitors of endothelial cell proliferation/migration, antagonists of angiogenic growth factors, inhibitors of Integrin/Survival signaling, and chelators of copper.
  • Methods of the invention may optionally further comprise a number of other steps.
  • the present methods further comprises the step of administering the optically functional MMP inhibitor into a bodily fluid of the subject.
  • the optically functional MMP inhibitor may be introduced into the patient by any suitable method, including intravenous, intraperitoneal or subcutaneous injection or infusion, oral administration, transdermal absorption through the skin, or by inhalation.
  • the method further comprises contacting a target tissue, such as an organ, tissue, tumor, lesion, or cell type, with a compound of any one of formulas (FX1 ) - (FX113) prior to or during the exposure step.
  • the method further comprises allowing the compound to accumulate in a target tissue prior to exposure of the optically functional MMP inhibitor to electromagnetic radiation.
  • the method further comprises targeting the diagnostic agent to a selected organ, tissue, tumor, lesion, inflammation, or cell type.
  • the optically functional MMP inhibitor is administered to the skin, a tumor, surgical site, or a wound site.
  • the optically functional MMP inhibitor is administered and/or delivered to a blood vessel, lung, heart, throat, ear, rectum, bladder, stomach, intestines, esophagus, liver, brain, prostrate, breast, or pancreas of the subject.
  • Example 4A Preparation of an optically functional pyrazine-containing MMP inhibitor having formula (FX106)
  • An optically functional MMP inhibitor of the invention having formula (FX106) was prepared via the following procedure.
  • a 100 ml RBF equipped with magnetic stir bar was charged with 1.0 gram (5.05 mmole) diacid, 704 mg ( 5.05 mmole) beta-alanine methyl ester HCI salt, 379 mg (5.05 mmole) t-butyl amine, 1.53 g (15.15 mmole) Triethylamine, 4.3 g (1 1.2 mmole) HATU in 75 dry DMF (HATU last) and stirred at room temperature overnight. The reaction was concentrated in vacuo and partionned between EA and aq. Citric acid. The organics were washed with satd. aq.
  • Example 4C Preparation of an optically functional pyrazine-containing MMP inhibitor having formula (FX108)
  • Example 4D Preparation of an optically functional indocyanine-containing MMP inhibitor having formula (FX105)
  • Figure 6A provides a high resolution mass spectrum of the compound having formula (FX105).
  • Figure 6B provides an expanded view of the mass spectrum (top panel) of the compound having formula (FX105) and a simulated mass spectrum (lower panel) for a mass to charge ratio (m/z) range of about 1 149 to about 1 154.
  • Figure 6C provides an expanded view of the mass spectrum (top panel) of the compound having formula (FX105) and a simulated mass spectrum (lower panel) for a mass to charge ratio (m/z) range of about 574 to 576.
  • Example 4E Optically functional azulene-containing MMP inhibitor having formulas (FX110) and (FX111)
  • the invention provides optically functional azulene-containing MMP inhibitors being of the formula (FX110):
  • the invention provides compounds having formula (FX110), wherein R is-OCF 3 .
  • invention provides optically functional azulene-containing MMP inhibitors being of the formula (FX111 ):
  • each of Z 1 - Z 3 is independently hydrogen or fluorine.
  • the invention provides compounds having formula (FX111 ), wherein each of Q 1 - Q 4 is independently hydrogen or -CN, -C0 2 R 40 , -S0 2 OR 43 , -CONR 52 R 53 , -COR 54 , -N0 2 , -SOR 41 , - S0 2 R 55 , -POsR ⁇ R 45 , halo, C C 6 acyl, trihalomethyl, or -S0 2 NR 56 R 57 .
  • the invention provides compounds having formula (FX111 ), wherein each of Q 1 - Q 4 is hydrogen, and wherein each of Z 1 - Z 3 is independently hydrogen or fluorine.
  • the invention provides compounds having formula (FX110), wherein R is-OCF 3 .
  • This class of azulene-containing MMP compounds may be prepared, for example, via peptide coupling to azulene acid. (See, e.g., Leermakers and Bowman. Structure vs reactivity in azulene carboxylic acids and esters. Journal of Organic Chemistry 1964, 29, 3708-371 1 .)
  • Example 4F Optically functional MMP inhibitors having an optical dye in the S1 ' binding pocket.
  • the invention provides optically function MMP inhibitors having an optical dye in the S1 ' binding pocket having the formula (FX112) or (FX113):
  • R and R ' is independently
  • each R and R ' is independently H or CrC 6 alkyl, and optionally wherein each R and R ' is independently H or C1-C3 alkyl.
  • Example 5 Compositions and methods for phototherapy
  • the invention includes phototherapy methods wherein a phototherapeutic agent comprising a compound of any one of the formulas (FX1 ) - (FX113) is administered to a patient, for example, wherein a therapeutically effective amount of such a component is administered to a patient in need of treatment.
  • compounds of the invention provide an optical agent capable of selectively targeting and delivery to tissue expressing MMPs and further functions as a phototherapeutic agent and MMP inhibitor.
  • the phototherapeutic agent is allowed to accumulate in a target region of interest (e.g., target tissue, tumor, or organ).
  • a target region of interest e.g., target tissue, tumor, or organ.
  • the phototherapeutic agent is activated by exposure to electromagnetic radiation.
  • the phototherapeutic agent is activated after an effective concentration of the phototherapeutic agent has accumulated in a target tissue.
  • An effective concentration of a compound of the invention depends on the nature of the formulation, method of delivery, target tissue, activation method and toxicity to the surrounding normal non-target tissue. Exposure to electromagnetic radiation and activation of the phototherapeutic agent may occur during or after administration of the phototherapeutic agent and accumulation at the target tissue.
  • the target region is illuminated with electromagnetic radiation having a wavelength in the range of about 350 nm to about 1300 nm, preferably for some applications in the range of about 350 nm to about 900 nm.
  • the wavelength of the electromagnetic radiation corresponds to a peak in the absorption spectrum of the phototherapeutic agent, for example is within 20 nanometers of a peak in the absorption spectrum of the phototherapeutic agent in the visible or NIR regions.
  • the target site is exposed to electromagnetic radiation having sufficient fluence and/or power sufficient to activate the phototherapeutic agent so as to induce cell death, for example via necrosis or apoptosis processes.
  • electromagnetic radiation of low energy, power or fluence is needed to activate the phototherapeutic agent. If the region of interest is, for example a lesion or tumor on the skin surface, the region can be directly illuminated. Otherwise, endoscopic and/or endoluminal catheters equipped with an
  • electromagnetic radiation source may be employed to provide a photodiagnostic and/or the phototherapeutic effect.
  • the fluence of the electromagentic radiation is preferably, but not always, kept below 200 mW/cm 2 to minimize undesirable thermal effects.
  • the intensity, power, and duration of the illumination, and the wavelength of the electromagnetic radiation may vary widely depending on the body location, the lesions site, the effect to be achieved, etc.
  • the power of the applied electromagnetic radiation is preferably selected over the range of 1 - 500 mW/cm 2 and optionally selected over the range of 1 - 200 mW/cm 2 .
  • the duration of the exposure to applied electromagentic radiation selected over the range of 1 seconds to 60 minutes.
  • the invention provides a method of using a phototherapeutic agent, the method comprising: (i) administering a therapeutically effective amount of a phototherapeutic agent to a subject, the phototherapeutic agent comprising a compound being of any one of formula (FX1 ) - (FX113), wherein at least one of X 1 to X 17 , R 1 to R 5 and R 8 to R 10 is PS 1 or PS 2 , or a pharmaceutically acceptable salt or ester thereof; and (ii) exposing the phototherapeutic agent administered to the patient to electromagnetic radiation.
  • erapy being of formula (FX1 ) or (FX2):
  • each PS 1 is independently an azide, azo, diazo, oxaza, diaza, dithia, thioxa, or dioxa group
  • each PS 2 is independently a group corresponding to a porphyrin, benzoporphyrin, phthalocyanine, phenothiazine, chlorin, bacteriochlorin, phthalocyanine, porphyrin, purpurin, merocyanine, pheophorbides, psoralen, aminolevulinic acid (ALA), hematoporphyrin
  • the invention provides compounds having formula (FX1 ) or (FX2) wherein at least one of R 1 - R 5 or A includes a Type 2 photosensitizer.
  • the invention provides compounds having formula (FX1 ) or (FX2) wherein at least one of R 1 - R 5 or A includes a Type 1 photosensitizer.
  • the invention provides a compound having formula (FX1 ) or (FX2) wherein R 1 and R 5 are each hydrogen.
  • the method of the invention comprises administering to a patient a compound having any one of formula selected from (FX1 ) - (FX113), including any of the specific compositions classes and compounds described in connection with formula (FX1 ) - (FX113), particularly those compounds having a photosensitizer component.
  • the phototherapeutic agent is exposed to a therapeutically effective amount of electromagnetic radiation.
  • a therapeutically effective amount of electromagentic radiation is an amount for achieving a desired therapeutic result, for example an amount for generating a therapeutically effective amount of reactive species for damaging or causing cell death of a selected target tissue.
  • the method further comprises generating one or more reactive species from said compound administered to the patient via the exposure of the phototherapeutic agent to applied electromagnetic radiation.
  • the method further comprises the step of cleaving one or more photolabile bonds of the optical agent so as to generate reactive species comprising free radicals.
  • the method further comprises targeting the phototherapeutic agent to a selected organ in the patient or to a selected tissue type in the patient.
  • a selected organ in the patient or to a selected tissue type in the patient.
  • a selected tissue type in the patient In an embodiment, a
  • therapeutically effective dose of the phototherapeutic agent is administered to a patient in need of treatment.
  • Embodiments of this aspect may comprise a method of carrying out an in vivo therapeutic and/or diagnostic procedure.
  • the invention comprises a method of carrying out an in vivo phototherapeutic, photoactivation, and/or photosensitizing procedure.
  • the present methods have broad clinical utility which includes, but is not limited to, phototherapy of tumors, inflammatory processes, and impaired vasculature.
  • subjects of the invention may be any mammal, such as a human, and optionally the subject of the present methods is a patient in need of treatment and/or diagnosis.
  • the present methods are also useful in ex vivo and in vitro procedures, including medical therapeutic and diagnostic procedures.
  • Methods of the invention may optionally further comprise a number of other steps.
  • the method further comprises the step of administering the phototherapeutic agent into a bodily fluid of the subject.
  • the phototherapeutic agent may be introduced into the patient by any suitable method, including intravenous, intraperitoneal or subcutaneous injection or infusion, oral administration, transdermal absorption through the skin, or by inhalation.
  • the method further comprises contacting a target tissue, such as an organ, tissue, tumor, lesion, or cell type, with a compound of any one of formulas (FX1 ) - (FX113) prior to or during the exposure step.
  • the method further comprises allowing the compound to accumulate in a target tissue prior to exposure of the phototherapeutic agent to electromagnetic radiation.
  • the method further comprises targeting the diagnostic agent to a selected organ, tissue, tumor, lesion, inflammation, or cell type.
  • the phototherapeutic agent is administered to the skin, a tumor, surgical site, or a wound site.
  • the phototherapeutic agent is administered and/or delivered to a blood vessel, lung, heart, throat, ear, rectum, bladder, stomach, intestines, esophagus, liver, brain, prostrate, breast, or pancreas of the subject.
  • the optical conditions for the step of exposing the phototherapeutic agent administered to the patient to electromagnetic radiation will vary considerably with the (i) therapeutic and/or diagnostic objectives, and (ii) the condition of the subject (e.g., height, weight, state of health etc.).
  • the applied condition of the subject e.g., height, weight, state of health etc.
  • electromagnetic radiation has wavelengths, energy and/or fluence sufficient to achieve a desired therapeutic and/or diagnostic result.
  • the electromagnetic radiation has wavelengths, energy and/or fluence sufficient to activate the phototherapeutic agent, for example wavelengths, energy and/or fluence sufficient to result in generation of reactive species, including singlet oxygen, ions, and/or free radicals.
  • the electromagnetic radiation has wavelengths, energy and/or fluence sufficient to result in cleavage of at least one photolabile bond of the optical agent upon absorption.
  • the electromagnetic radiation exposed to the phototherapeutic agent has wavelengths
  • excitation is achieved using electromagnetic substantially free (e.g., less than about 10% of total radiant energy), of ultraviolet radiation, for example, to minimize exposure of the subject to electromagnetic radiation capable of causing unwanted cell or tissue damage.
  • Electromagnetic radiation may be provided to the phototherapeutic agent using a range of optical sources and/or surgical instrumentation, including a laser, light emitting diodes, fiber optic device, endoscope, catheter, optical filters, or any combination of these.
  • the invention contemplates pharmaceutically active compounds either chemically synthesized or formed by in vivo biotransformation to compounds set forth herein.
  • compositions of the invention is a compound or salt or ester thereof suitable for pharmaceutical formulations.
  • Prodrugs of the compounds of the invention are useful in embodiments including compositions and methods. Any compound that will be converted in vivo to provide a biologically, pharmaceutically, diagnostically, or therapeutically active form of a compound of the invention is a prodrug.
  • Various examples and forms of prodrugs are well known in the art. Examples of prodrugs are found, inter alia, in: Design of Prodrugs, edited by H. Bundgaard, (Elsevier, 1985); Methods in Enzymology, Vol. 42, at pp. 309-396, edited by K. Widder, et. al.
  • a prodrug such as a pharmaceutically acceptable prodrug
  • Prodrugs of the invention can be rapidly transformed in vivo to a parent compound of a compound described herein, for example, by hydrolysis in blood or by other cell, tissue, organ, or system processes. Further discussion is provided in: T. Higuchi and V. Stella, Pro-drugs as Novel Delivery Systems, V. 14 of the A.C.S. Symposium Series; and in Edward B. Roche, ed., Bioreversible Carriers in Drug Design, American Pharmaceutical Association and Pergamon Press (1987).
  • Optical agents of the invention can be formulated with pharmaceutically-acceptable anions and/or cations.
  • Pharmaceutically-acceptable cations include among others, alkali metal cations (e.g., Li + , Na + , K + ), alkaline earth metal cations (e.g., Ca 2+ , Mg 2+ ), non-toxic heavy metal cations and ammonium (NH 4 + ) and substituted ammonium (N(R') 4 + , where R' is hydrogen, alkyl, or substituted alkyl, i.e., including, methyl, ethyl, or hydroxyethyl, specifically, trimethyl ammonium, triethyl ammonium, and triethanol ammonium cations).
  • alkali metal cations e.g., Li + , Na + , K +
  • alkaline earth metal cations e.g., Ca 2+ , Mg 2+
  • Pharmaceutically-acceptable anions include, among others, halides (e.g., F “ , CI “ , Br “ , At “ ), sulfate, acetates (e.g., acetate, trifluoroacetate), ascorbates, aspartates, benzoates, citrates, and lactate.
  • Pharmaceutically acceptable salts comprise pharmaceutically-acceptable anions and/or cations.
  • pharmaceutically acceptable salt can refer to acid addition salts or base addition salts of the compounds in the present disclosure.
  • a pharmaceutically acceptable salt is any salt which retains at least a portion of the activity of the parent compound and does not impart significant deleterious or undesirable effect on a subject to whom it is administered and in the context in which it is administered.
  • Pharmaceutically acceptable salts include metal complexes and salts of both inorganic and organic acids.
  • Pharmaceutically acceptable salts include metal salts such as aluminum, calcium, iron, magnesium, manganese and complex salts.
  • salts include, but are not limited to, acid salts such as acetic, aspartic, alkylsulfonic, arylsulfonic, axetil, benzenesulfonic, benzoic, bicarbonic, bisulfuric, bitartaric, butyric, calcium edetate, camsylic, carbonic, chlorobenzoic, cilexetil, citric, edetic, edisylic, estolic, esyl, esylic, formic, fumaric, gluceptic, gluconic, glutamic, glycolic, glycolylarsanilic, hexamic, hexylresorcjnoic, hydrabamic, hydrobromic, hydrochloric, hydroiodic, hydroxynaphthoic, isethionic, lactic, lactobionic, maleic, malic, malonic, mandelic, methanesulfonic, methylnitric
  • Pharmaceutically acceptable salts can be derived from amino acids, including, but not limited to, cysteine.
  • Other pharmaceutically acceptable salts can be found, for example, in Stahl et al., Handbook of Pharmaceutical Salts: Properties, Selection, and Use, Wiley-VCH, Verlag Helvetica Chimica Acta, Zurich, 2002. (ISBN 3-906390-26-8).
  • a compound of the invention, or pharmaceutically acceptable salt thereof is administered to a subject in a diagnostically or therapeutically effective amount.
  • a diagnostically or therapeutically effective amount One skilled in the art generally can determine an appropriate dosage.
  • compositions for oral administration can be, for example, prepared in a manner such that a single dose in one or more oral preparations contains at least about 20 mg of the present compound per square meter of subject body surface area, or at least about 50, 100, 150, 200, 300, 400, or 500 mg of the present compound per square meter of subject body surface area (the average body surface area for a human is, for example, 1.8 square meters).
  • a single dose of a composition for oral administration can contain from about 20 to about 600 mg, and in certain aspects from about 20 to about 400 mg, in another aspect from about 20 to about 300 mg, and in yet another aspect from about 20 to about 200 mg of the present compound per square meter of subject body surface area.
  • compositions for parenteral administration can be prepared in a manner such that a single dose contains at least about 20 mg of the present compound per square meter of subject body surface area, or at least about 40, 50, 100, 150, 200, 300, 400, or 500 mg of the present compound per square meter of subject body surface area.
  • a single dose in one or more parenteral preparations contains from about 20 to about 500 mg, and in certain aspects from about 20 to about 400 mg, and in another aspect from about 20 to about 450 mg, and in yet another aspect from about 20 to about 350 mg of the present compound per square meter of subject body surface area.
  • these oral and parenteral dosage ranges represent generally preferred dosage ranges, and are not intended to limit the invention.
  • the dosage regimen actually employed can vary widely, and, therefore, can deviate from the generally preferred dosage regimen. It is contemplated that one skilled in the art will tailor these ranges to the individual subject.
  • Toxicity and therapeutic efficacy of such compounds and bioconjugates can be determined by standard pharmaceutical procedures in cell cultures or experimental animals for determining the LD 50 (the dose lethal to 50% of the population) and the ED 50 , (the dose therapeutically effective in 50% of the population).
  • the dose ratio between toxic and therapeutic effects is the therapeutic index that can be expressed as the ratio LD 5 o/ED 5 o.
  • Compounds and bioconjugates that exhibit large therapeutic indices are preferred. While compounds and bioconjugates exhibiting toxic side effects can be used, care should be taken to design a delivery system that targets such compounds and bioconjugates to the site affected by the disease or disorder in order to minimize potential damage to unaffected cells and reduce side effects.
  • the dosage of such compounds and bioconjugates lies preferably within a range of circulating plasma or other bodily fluid concentrations that include the ED 50 and provides clinically efficacious results (i.e., reduction in disease symptoms).
  • the dosage can vary within this range depending upon the dosage form employed and the route of administration utilized.
  • the therapeutically effective amount can be estimated initially from cell culture assays.
  • a dosage can be formulated in animal models to achieve a circulating plasma concentration range that includes the ED 50 (the concentration of the test compound that achieves a half-maximal inhibition of symptoms) as determined in cell culture. Such information can be used to more accurately determine useful dosages in humans and other mammals.
  • Compound and bioconjugate levels in plasma can be measured, for example, by high performance liquid chromatography.
  • An amount of a compound or bioconjugate that can be combined with a pharmaceutically acceptable carrier to produce a single dosage form will vary depending upon the patient treated and the particular mode of administration. It will be appreciated by those skilled in the art that the unit content of a compound/bioconjugate contained in an individual dose of each dosage form need not in itself constitute a therapeutically effective amount, as the necessary therapeutically effective amount could be reached by administration of a number of individual doses. The selection of dosage depends upon the dosage form utilized, the condition being treated, and the particular purpose to be achieved according to the determination of those skilled in the art.
  • the dosage and dosage regime for treating a disease or condition can be selected in accordance with a variety of factors, including the type, age, weight, sex, diet and/or medical condition of the patient, the route of administration, pharmacological considerations such as activity, efficacy, pharmacokinetic and/or toxicology profiles of the particular compound/bioconjugate employed, whether a compound/bioconjugate delivery system is utilized, and/or whether the compound/bioconjugate is administered as a pro-drug or part of a drug combination.
  • the dosage regime actually employed can vary widely from subject to subject, or disease to disease and different routes of administration can be employed in different clinical settings.
  • compositions/formulations of the present invention comprise a therapeutically effective amount (which can optionally include a diagnostically effective amount) of at least one compound or bioconjugate of the present invention.
  • Subjects receiving treatment that includes a compound/bioconjugate of the invention are preferably animals (e.g., mammals, reptiles and/or avians), more preferably humans, horses, cows, dogs, cats, sheep, pigs, and/or chickens, and most preferably humans.
  • the preferred composition depends on the route of administration. Any route of administration can be used as long as the target of the compound or pharmaceutically acceptable salt is available via that route. Suitable routes of administration include, for example, oral, intravenous, parenteral, inhalation, rectal, nasal, topical (e.g., transdermal and intraocular), intravesical, intrathecal, enteral, pulmonary, intralymphatic, intracavital, vaginal, transurethral, intradermal, aural, intramammary, buccal, orthotopic, intratracheal, intralesional, percutaneous, endoscopical, transmucosal, sublingual, and intestinal administration.
  • routes of administration include, for example, oral, intravenous, parenteral, inhalation, rectal, nasal, topical (e.g., transdermal and intraocular), intravesical, intrathecal, enteral, pulmonary, intralymphatic, intracavital, vaginal, transurethral, intradermal, aural, intramamm
  • the invention provides a method for treating a medical condition comprising administering to a subject (e.g. patient) in need thereof, a therapeutically effective amount of a composition of the invention, such as a compound of any one of formulas (FX1 ) - (FX113).
  • a composition of the invention such as a compound of any one of formulas (FX1 ) - (FX113).
  • the invention provides a method for diagnosing or aiding in the diagnosis of a medical condition comprising administering to a subject in need thereof, a diagnostically effective amount of a composition of the invention.
  • the medical condition is cancer, or various other diseases, injuries, and disorders, including cardiovascular disorders such as atherosclerosis and vascular restenosis, inflammatory diseases, ophthalmic diseases and dermatological diseases.
  • the diagnostic and therapeutic formulations of this invention can be administered alone, but can be administered with a pharmaceutical carrier selected upon the basis of the chosen route of administration and standard pharmaceutical practice.
  • diagnostic and therapeutic formulations of the invention can be administered intravenously, in oral dosage forms, intraperitoneally, subcutaneously, or intramuscularly, all using dosage forms well known to those of ordinary skill in the pharmaceutical arts.
  • compositions, preparations and formulations can be formulated into diagnostic or therapeutic compositions for enteral, parenteral, topical, aerosol, inhalation, or cutaneous administration.
  • Topical or cutaneous delivery of the compositions, preparations and formulations can also include aerosol formulation, creams, gels, solutions, etc.
  • the present compositions, preparations and formulations are administered in doses effective to achieve the desired diagnostic and/or therapeutic effect. Such doses can vary widely depending upon the particular compositions employed in the composition, the organs or tissues to be examined, the equipment employed in the clinical procedure, the efficacy of the treatment achieved, and the like.
  • These compositions, preparations and formulations contain an effective amount of the composition(s), along with conventional pharmaceutical carriers and excipients appropriate for the type of administration contemplated.
  • These compositions, preparations and formulations can also optionally include stabilizing agents and skin penetration enhancing agents.
  • Compounds and bioconjugates of the present invention can be formulated for parenteral administration by injection (e.g., by bolus injection or continuous infusion).
  • Formulations for injection can be presented in unit dosage form in ampoules or in multi-dose containers with an optional preservative added.
  • the parenteral preparation can be enclosed in ampoules, disposable syringes or multiple dose vials made of glass, plastic or the like.
  • the formulation can take such forms as suspensions, solutions or emulsions in oily or aqueous vehicles, and can contain formulatory agents such as suspending, stabilizing and/or dispersing agents.
  • a parenteral preparation can be a sterile injectable solution or suspension in a nontoxic parenterally acceptable diluent or solvent (e.g., as a solution in 1 ,3-butanediol).
  • a nontoxic parenterally acceptable diluent or solvent e.g., as a solution in 1 ,3-butanediol.
  • acceptable vehicles and solvents that can be employed are water, Ringer's solution, and isotonic sodium chloride solution.
  • sterile, fixed oils are conventionally employed as a solvent or suspending medium.
  • any bland fixed oil can be employed including synthetic mono- or di-glycerides.
  • fatty acids such as oleic acid can be used in the parenteral preparation.
  • compounds and bioconjugates of the present invention can be formulated in powder form for constitution with a suitable vehicle, such as sterile pyrogen-free water, before use.
  • a compound/bioconjugate suitable for parenteral administration can include a sterile isotonic saline solution containing between 0.1 percent and 90 percent weight per volume of the compound/bioconjugate.
  • a solution can contain from about 5 percent to about 20 percent, more preferably from about 5 percent to about 17 percent, more preferably from about 8 to about 14 percent, and still more preferably about 10 percent weight per volume of the compound/bioconjugate.
  • the solution or powder preparation can also include a solubilizing agent and a local anesthetic such as lignocaine to ease pain at the site of the injection.
  • a solubilizing agent such as lignocaine
  • a local anesthetic such as lignocaine
  • a compound/bioconjugate of the invention can be formulated to take the form of tablets or capsules prepared by conventional means with one or more pharmaceutically acceptable carriers (e.g., excipients such as binding agents, fillers, lubricants and disintegrants).
  • pharmaceutically acceptable carriers e.g., excipients such as binding agents, fillers, lubricants and disintegrants.
  • Controlled-release (or sustained-release) preparations can be formulated to extend the activity of a compound/bioconjugate and reduce dosage frequency. Controlled-release preparations can also be used to effect the time of onset of action or other characteristics, such as blood levels of the compound/bioconjugate, and consequently affect the occurrence of side effects.
  • Controlled-release preparations can be designed to initially release an amount of a compound/bioconjugate that produces the desired therapeutic effect, and gradually and continually release other amounts of the compound/bioconjugate to maintain the level of therapeutic effect over an extended period of time.
  • the compound/bioconjugate can be released from the dosage form at a rate that will replace the amount of compound/bioconjugate being metabolized and/or excreted from the body.
  • the controlled-release of a compound/bioconjugate can be stimulated by various inducers, e.g., change in pH, change in temperature, enzymes, water, and/or other physiological conditions or molecules.
  • Controlled-release systems can include, for example, an infusion pump which can be used to administer the compound/bioconjugate in a manner similar to that used for delivering insulin or chemotherapy to the body generally, or to specific organs or tumors.
  • the compound/bioconjugate is administered in combination with a biodegradable, biocompatible polymeric implant that releases the compound/bioconjugate over a controlled period of time at a selected site.
  • polymeric materials include polyanhydrides, polyorthoesters, polyglycolic acid, polylactic acid, polyethylene vinyl acetate, and copolymers and combinations thereof.
  • a controlled release system can be placed in proximity of a therapeutic target (e.g., organ, tissue, or group of cells), thus requiring only a fraction of a systemic dosage.
  • Compounds/bioconjugates of the invention can be administered by other controlled- release means or delivery devices that are well known to those of ordinary skill in the art. These include, for example, hydropropylmethyl cellulose, other polymer matrices, gels, permeable membranes, osmotic systems, multilayer coatings, microparticles, liposomes, microspheres, or the like, or a combination of any of the above to provide the desired release profile in varying proportions. Other methods of controlled-release delivery of compounds/bioconjugates will be known to the skilled artisan and are within the scope of the invention,
  • Compounds/bioconjugates of the invention can be administered directly to the lung of a patient/subject by inhalation.
  • a compound/bioconjugate can be conveniently delivered to the lung by a number of different devices.
  • a Metered Dose Inhaler which utilizes canisters that contain a suitable low boiling point propellant, e.g., dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas can be used to deliver a compound/bioconjugate directly to the lung.
  • MDI devices are available from a number of suppliers such as 3M Corporation, Aventis, Boehringer Ingleheim, Forest Laboratories, GlaxoSmithKline, Merck & Co. and Vectura.
  • a Dry Powder Inhaler (DPI) device can be used to administer a compound/bioconjugate to the lung.
  • DPI devices typically use a mechanism such as a burst of gas to create a cloud of dry powder inside a container, which can then be inhaled by the patient.
  • DPI devices are also well known in the art and can be purchased from a number of vendors which include, for example, GlaxoSmithKline, Nektar Therapeutics, Innovata and Vectura.
  • MDDPI multiple dose DPI
  • a popular variation is the multiple dose DPI (“MDDPI”) system, which allows for the delivery of more than one therapeutic dose.
  • MDDPI devices are available from companies such as AstraZeneca, GlaxoSmithKline, TEVA, Merck & Co., SkyePharma and Vectura.
  • capsules and cartridges of gelatin for use in an inhaler or insufflator can be formulated containing a powder mix of the compound/bioconjugate and a suitable powder base such as lactose or starch for these systems.
  • a liquid spray device supplied, for example, by Aradigm Corporation.
  • Liquid spray systems use extremely small nozzle holes to aerosolize liquid compound/bioconjugate formulations that can then be directly inhaled into the lung.
  • a nebulizer device can be used to deliver a compound/bioconjugate to the lung.
  • Nebulizers create aerosols from liquid compound/bioconjugate formulations by using, for example, ultrasonic energy to form fine particles that can be readily inhaled. Examples of nebulizers include devices supplied by Aventis and Battelle.
  • an electrohydrodynamic (“EHD”) aerosol device can be used to deliver a compound/bioconjugate to the lung.
  • EHD aerosol devices use electrical energy to aerosolize liquid compound/bioconjugate solutions or suspensions.
  • the electrochemical properties of the compound/bioconjugate formulation are important parameters to optimize when delivering this compound/bioconjugate to the lung with an EHD aerosol device. Such optimization is routinely performed by one of skill in the art.
  • Other methods of intra-pulmonary delivery of compounds/bioconjugates will be known to the skilled artisan and are within the scope of the invention.
  • Liquid compound/bioconjugate formulations suitable for use with nebulizers and liquid spray devices and EHD aerosol devices will typically include the compound/bioconjugate with a pharmaceutically acceptable carrier.
  • the pharmaceutically acceptable carrier is a liquid such as alcohol, water, polyethylene glycol or a perfluorocarbon.
  • another material can be added to alter the aerosol properties of the solution or suspension of the compound/bioconjugate.
  • this material can be a liquid such as an alcohol, glycol, polyglycol or a fatty acid.
  • Other methods of formulating liquid compound/bioconjugate solutions or suspensions suitable for use in aerosol devices are known to those of skill in the art.
  • a compound/bioconjugate of the invention can be formulated as a depot preparation. Such long-acting formulations can be administered by implantation (e.g., subcutaneously or intramuscularly) or by intramuscular injection. Accordingly, the compound/bioconjugate can be formulated with suitable polymeric or hydrophobic materials such as an emulsion in an acceptable oil or ion exchange resin, or as sparingly soluble derivatives such as a sparingly soluble salt. Other methods of depot delivery of compounds/bioconjugates will be known to the skilled artisan and are within the scope of the invention.
  • a compound/bioconjugate can be combined with a pharmaceutically acceptable carrier so that an effective dosage is delivered, based on the desired activity ranging from an effective dosage, for example, of 1 .0 ⁇ to 1 .0 mM.
  • a topical formulation of a compound/bioconjugate can be applied to the skin.
  • the pharmaceutically acceptable carrier can be in the form of, for example, and not by way of limitation, an ointment, cream, gel, paste, foam, aerosol, suppository, pad or gelled stick.
  • a topical formulation can include a therapeutically effective amount of a compound/bioconjugate in an ophthalmologically acceptable excipient such as buffered saline, mineral oil, vegetable oils such as corn or arachis oil, petroleum jelly, Miglyol 182, alcohol solutions, or liposomes or liposome-like products.
  • an ophthalmologically acceptable excipient such as buffered saline, mineral oil, vegetable oils such as corn or arachis oil, petroleum jelly, Miglyol 182, alcohol solutions, or liposomes or liposome-like products.
  • ophthalmologically acceptable excipient such as buffered saline, mineral oil, vegetable oils such as corn or arachis oil, petroleum jelly, Miglyol 182, alcohol solutions, or liposomes or liposome-like products.
  • Any of these formulations of such compounds/bioconjugates can include preservatives, antioxidants, antibiotics, immunosuppressants, and other biologically or pharmaceutically effective agents that do
  • Compounds/bioconjugates of the invention can be formulated in rectal formulations such as suppositories or retention enemas that include conventional suppository bases such as cocoa butter or other glycerides and/or binders and/or carriers such as triglycerides, microcrystalline cellulose, gum tragacanth or gelatin. Rectal formulations can contain a compound/bioconjugate in the range of 0.5% to 10% by weight, for example. Other methods of rectal delivery of compounds/bioconjugates will be known to the skilled artisan and are within the scope of the invention.
  • the invention provides a medicament which comprises a therapeutically effective amount of one or more compositions of the invention, such as a compound of any one of formulas (FX1 ) - (FX113).
  • the invention provides a medicament which comprises a diagnostically effective amount of one or more compositions of the invention.
  • the invention provides a method for making a medicament for treatment of a condition described herein, such as the treatment of cancer, inflammation, stenosis or a vascular disease.
  • the invention provides a method for making a medicament for diagnosis or aiding in the diagnosis of a condition described herein, such as the diagnosis of cancer, inflammation, stenosis or a vascular disease.
  • the invention provides the use of one or more compositions set forth herein for the making of a medicament for the treatment of cancer, inflammation, stenosis or a vascular disease.
  • the invention provides the use of one or more compositions set forth herein for the treatment of a disease.
  • the invention provides the use of one or more compositions set forth herein for the diagnosis of a disease.
  • Compositions of the invention include formulations and preparations comprising one or more of the present optical agents provided in an aqueous solution, such as a pharmaceutically acceptable formulation or preparation.
  • compositions of the invention further comprise one or more pharmaceutically acceptable surfactants, buffers, electrolytes, salts, carriers, binders, coatings, preservatives and/or excipients.
  • the invention provides a pharmaceutical formulation having an active ingredient comprising a composition of the invention, such as a compound of any one of formulas (FX1 ) - (FX113).
  • the invention provides a method of synthesizing a composition of the invention or a pharmaceutical formulation thereof, such as a compound of any one of formulas (FX1 ) - (FX113).
  • a pharmaceutical formulation comprises one or more excipients, carriers, diluents, and/or other components as would be understood in the art.
  • the components meet the standards of the National Formulary ("IMF”), United States Pharmacopoeia (“USP”; United States Pharmacopeial Convention Inc., Rockville, Maryland), or Handbook of Pharmaceutical Manufacturing Formulations (Sarfaraz K. Niazi, all volumes, ISBN: 9780849317521 , ISBN 10: 0849317525; CRC Press, 2004).
  • IMF National Formulary
  • USP United States Pharmacopoeia
  • USP United States Pharmacopeial Convention Inc., Rockville, Maryland
  • Handbook of Pharmaceutical Manufacturing Formulations (Sarfaraz K. Niazi, all volumes, ISBN: 9780849317521 , ISBN 10: 0849317525; CRC Press, 2004).
  • the formulation base of the formulations of the invention comprises physiologically acceptable excipients, namely, at least one binder and optionally other physiologically acceptable excipients.
  • physiologically acceptable excipients are those known to be usable in the pharmaceutical technology sectors and adjacent areas, particularly, those listed in relevant pharmacopeias (e.g. DAB, Ph. Eur., BP, NF, USP), as well as other excipients whose properties do not impair a physiological use.
  • compositions including a therapeutically effective amount of a compound or salt of this invention, as well as processes for making such compositions.
  • Such compositions generally include one or more pharmaceutically acceptable carriers (e.g., excipients, vehicles, auxiliaries, adjuvants, diluents) and can include other active ingredients.
  • pharmaceutically acceptable carriers e.g., excipients, vehicles, auxiliaries, adjuvants, diluents
  • Formulation of these compositions can be achieved by various methods known in the art. A general discussion of these methods can be found in, for example, Hoover, John E., Remington's Pharmaceutical Sciences (Mack Publishing Co., Easton, PA: 1975). See also, Lachman, L, eds., Pharmaceutical Dosage Forms (Marcel Decker, New York, N. Y., 1980).
  • compositions and medicaments of this invention can further comprise one or more pharmaceutically acceptable carriers, excipients, buffers, emulsifiers, surfactants, electrolytes or diluents.
  • pharmaceutically acceptable carriers such as, for example, those described in Remingtons Pharmaceutical Sciences, 17th edition, ed. Alfonoso R. Gennaro, Mack Publishing Company, Easton, Pa. (1985).
  • compositions of the invention include formulations and preparations comprising one or more of the present compounds provided in an aqueous solution, such as a pharmaceutically acceptable formulation or preparation.
  • compositions of the invention further comprise one or more pharmaceutically acceptable surfactants, buffers, electrolytes, salts, carriers, binders, coatings, preservatives and/or excipients.
  • Compounds and bioconjugates of the present invention can be formulated by known methods for administration to a subject using several routes which include, but are not limited to, parenteral, oral, topical, intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal, epidural, and ophthalmic routes.
  • An individual compound/bioconjugate can be administered in combination with one or more additional compounds/bioconjugates of the present invention and/or together with other biologically active or biologically inert agents.
  • Such biologically active or inert agents can be in fluid or mechanical communication with the compound(s)/bioconjugate(s) or attached to the compound(s)/bioconjugate(s) by ionic, covalent, Van der Waals, hydrophobic, hydrophilic or other physical forces. It is preferred that administration is localized in a subject, but administration can also be systemic.
  • Compounds and bioconjugates of the present invention can be formulated by any conventional manner using one or more pharmaceutically acceptable carriers.
  • the compound(s)/bioconjugate(s) and their pharmaceutically acceptable salts and solvates can be specifically formulated for administration, e.g., by inhalation or insufflation (either through the mouth or the nose) or oral, buccal, parenteral or rectal administration.
  • the compounds/bioconjugates can take the form of charged, neutral and/or other pharmaceutically acceptable salt forms.
  • pharmaceutically acceptable carriers include, but are not limited to, those described in REMINGTON'S PHARMACEUTICAL SCIENCES (A.R. Gennaro, Ed.), 20th edition, Williams & Wilkins PA, USA (2000).
  • Compounds and bioconjugates of the present invention can be formulated in the form of solutions, suspensions, emulsions, tablets, pills, capsules, powders, controlled- or sustained- release formulations and the like.
  • Such formulations will contain a therapeutically effective amount of the compound/bioconjugate, preferably in purified form, together with a suitable amount of carrier so as to provide the form for proper administration to the patient.
  • the formulation should suit the mode of administration.
  • Pharmaceutically acceptable carriers that can be used in conjunction with the compounds of the invention are well known to those of ordinary skill in the art. Carriers can be selected based on a number of factors including, for example, the particular compound(s) or pharmaceutically acceptable salt(s) used; the compound's concentration, stability, and intended bioavailability; the condition being treated; the subject's age, size, and general condition; the route of administration; etc. A general discussion related to carriers can be found in, for example, J.G. Nairn, Remington's Pharmaceutical Science, pp. 1492-1517 (A. Gennaro, ed., Mack Publishing Co., Easton, Pa. (1985)).
  • Solid dosage forms for oral administration include, for example, capsules, tablets, gelcaps, pills, dragees, troches, powders, granules, and lozenges.
  • the compounds or pharmaceutically acceptable salts thereof can be combined with one or more pharmaceutically acceptable carriers.
  • the compounds and pharmaceutically acceptable salts thereof can be mixed with carriers including, but not limited to, lactose, sucrose, starch powder, corn starch, potato starch, magnesium carbonate, microcrystalline cellulose, cellulose esters of alkanoic acids, cellulose alkyl esters, talc, stearic acid, magnesium stearate, magnesium oxide, sodium and calcium salts of phosphoric and sulfuric acids, sodium carbonate, agar, mannitol, sorbitol, sodium saccharin, gelatin, acacia gum, alginic acid, sodium alginate, tragacanth, colloidal silicon dioxide, croscarmellose sodium, polyvinylpyrrolidone, and/or polyvinyl alcohol, and then tableted or encapsulated for convenient administration.
  • carriers including, but not limited to, lactose, sucrose, starch powder, corn starch, potato starch, magnesium carbonate, microcrystalline cellulose, cellulose esters of alkanoic acids, cellulose alkyl est
  • Such capsules or tablets can contain a controlled-release formulation, as can be provided in a dispersion of the compound or salt in hydroxypropylmethyl cellulose.
  • the dosage forms also can include buffering agents, such as sodium citrate, or magnesium or calcium carbonate or bicarbonate. Tablets and pills additionally can, for example, include a coating (e.g., an enteric coating) to delay disintegration and absorption.
  • the concentration of the present compounds in a solid oral dosage form can be from about 5 to about 50% for example, and in certain aspects from about 8 to about 40%, and in another aspect from about 10 to about 30% by weight based on the total weight of the composition.
  • Liquid dosage forms of the compounds of the invention for oral administration include, for example, pharmaceutically acceptable emulsions, solutions, suspensions, syrups, and elixirs containing inert diluents commonly used in the art (e.g., water). Such compositions also can include adjuvants, such as wetting, emulsifying, suspending, flavoring (e.g., sweetening), and/or perfuming agents.
  • the concentration of the present compounds in the liquid dosage form can be from about 0.01 to about 5 mg, and in certain aspects from about 0.01 to about 1 mg, and in another aspect from about 0.01 to about 0.5 mg per ml of the composition.
  • Low concentrations of the compounds of the invention in liquid dosage form can be prepared in the case that the compound is more soluble at low concentrations.
  • Techniques for making oral dosage forms useful in the invention are generally described in, for example, Modern Pharmaceutics, Chapters 9 and 10 (Banker & Rhodes, Editors (1979)). See also, Lieberman et al., Pharmaceutical Dosage Forms: Tablets (1981 ). See also, Ansel, Introduction to Pharmaceutical Dosage Forms (2nd Edition (1976)).
  • tablets or powders for oral administration can be prepared by dissolving the compound in a pharmaceutically acceptable solvent capable of dissolving the compound to form a solution and then evaporating when the solution is dried under vacuum.
  • a carrier can also be added to the solution before drying.
  • the resulting solution can be dried under vacuum to form a glass.
  • the glass can then be mixed with a binder to form a powder.
  • This powder can be mixed with fillers or other conventional tableting agents, and then processed to form a tablet.
  • the powder can be added to a liquid carrier to form a solution, emulsion, suspension, or the like.
  • solutions for oral administration are prepared by dissolving the compound in a pharmaceutically acceptable solvent capable of dissolving the compound to form a solution.
  • An appropriate volume of a carrier is added to the solution while stirring to form a pharmaceutically acceptable solution for oral administration.
  • a liposome or micelle can be utilized as a carrier or vehicle for the composition.
  • the compound can be a part of the lipophilic bilayers or micelle, and the targeting ligand, if present, can be on the external surface of the liposome or micelle.
  • a targeting ligand can be externally attached to the liposome or micelle after formulation for targeting the liposome or micelle (which contains the optical agents) to the desired tissue, organ, or other site in the body.
  • Injectable preparations can be formulated according to the known art using suitable dispersing, wetting agents, and/or suspending agents.
  • Acceptable vehicles for parenteral use include both aqueous and nonaqueous pharmaceutically-acceptable solvents.
  • Suitable pharmaceutically acceptable aqueous solvents include, for example, water, saline solutions, dextrose solutions (such as DW5), electrolyte solutions, etc.
  • the present compounds are formulated as nanoparticles or microparticles.
  • Use of such nanoparticle or microparticle formulations can be beneficial for some applications to enhance delivery, localization, target specificity, administration, etc. of the compound.
  • Potentially useful nanoparticles and microparticles include, but are not limited to, micelles, liposomes, microemulsions, nanoemulsions, vesicles, tubular micelles, cylindrical micelles, bilayers, folded sheets structures, globular aggregates, swollen micelles, inclusion complex, encapsulated droplets, microcapsules, nanocapsules or the like.
  • the present compounds can be located inside the nanoparticle or microparticle, within a membrane or wall of the nanoparticle or microparticle, or outside of (but bonded to or otherwise associated with) the nanoparticle or microparticle.
  • the agent formulated in nanoparticles or microparticles can be administered by any of the routes previously described. In a formulation applied topically, the compound is slowly released over time. In an injectable formulation, the liposome, micelle, capsule, etc., circulates in the bloodstream and is delivered to the desired site (e.g., target tissue).
  • liposomes can be prepared from dipalmitoyi phosphatidylcholine (DPPC) or egg phosphatidylcholine (PC) because this lipid has a low heat transition.
  • DPPC dipalmitoyi phosphatidylcholine
  • PC egg phosphatidylcholine
  • Liposomes are made using standard procedures as known to one skilled in the art (e.g., Braun-Falco et al., (Eds.), Griesbach Conference, Liposome Dermatics, Springer-Verlag, Berlin (1992), pp. 69 81 ; 91 1 17.
  • Polycaprolactone, poly(glycolic) acid, poly(lactic) acid, polyanhydride or lipids can be formulated as microspheres.
  • the present compounds can be mixed with polyvinyl alcohol (PVA), the mixture then dried and coated with ethylene vinyl acetate, then cooled again with PVA.
  • PVA polyvinyl alcohol
  • the present compounds can be within one or both lipid bilayers, in the aqueous between the bilayers, or within the center or core.
  • Liposomes can be modified with other molecules and lipids to form a cationic liposome. Liposomes can also be modified with lipids to render their surface more hydrophilic which increases their circulation time in the bloodstream.
  • the thus-modified liposome has been termed a "stealth" liposome, or a long-lived liposome, as described in U.S. Pat. No. 6,258,378, and in Stealth Liposomes, Lasic and Martin (Eds.) 1995 CRC Press, London.
  • Encapsulation methods include detergent dialysis, freeze drying, film forming, injection, as known to one skilled in the art and disclosed in, for example, U.S. Pat. No. 6,406,713.
  • compositions and methods include a micelle delivery system, for example, involving one or more PEG-based amphiphilic polymers developed for drug delivery including: PEG-poly(s-caprolactone), PEG-poly(amino acid), PEG- polylactide or PEG-phospholipid constructs; a cross linked poly(acrylic acid) polymer system, a phospholipid-based system and/or block copolymer systems comprising one or more of the following polymer blocks: a poly(lactic acid) polymer block; a poly(propylene glycol) polymer block; a poly(amino acid) polymer block; a poly(ester) polymer block; a poly ( ⁇ -caprolactone) polymer block; a poly(ethylene glycol) block, a poly(acrylic acid) block; a polylactide block; a polyester block; a polyamide block; a polyanhydride block; a polyurethane block; a polyimine block; a polyurea
  • Suitable pharmaceutically-acceptable nonaqueous solvents include, but are not limited to, the following (as well as mixtures thereof):
  • Alcohols (these include, for example, o-glycerol formal, ⁇ -glycerol formal, 1 , 3- butyleneglycol, aliphatic or aromatic alcohols having from 2 to about 30 carbons (e.g., methanol, ethanol, propanol, isopropanol, butanol, t-butanol, hexanol, octanol, amylene hydrate, benzyl alcohol, glycerin (glycerol), glycol, hexylene, glycol, tetrahydrofuranyl alcohol, cetyl alcohol, and stearyl alcohol), fatty acid esters of fatty alcohols (e.g., polyalkylene glycols, such as polypropylene glycol and polyethylene glycol), sorbitan, sucrose, and cholesterol);
  • o-glycerol formal e.g., o-glycerol formal, ⁇ -glycerol formal, 1 , 3- buty
  • Amides which include, for example, dimethylacetamide (DMA), benzyl benzoate DMA, dimethylformamide, N-hydroxyethyO-lactamide, N, N-dimethylacetamide-amides, 2- pyrrolidinone, 1-methyl-2-pyrrolidinone, and polyvinylpyrrolidone;
  • DMA dimethylacetamide
  • benzyl benzoate DMA dimethylformamide
  • N-hydroxyethyO-lactamide N, N-dimethylacetamide-amides
  • 2- pyrrolidinone 1-methyl-2-pyrrolidinone
  • polyvinylpyrrolidone polyvinylpyrrolidone
  • Esters which include, for example, acetate esters (e.g., monoacetin, diacetin, and triacetin), aliphatic and aromatic esters (e.g., ethyl caprylate or octanoate, alkyl oleate, benzyl benzoate, or benzyl acetate), dimethylsulfoxide (DMSO), esters of glycerin (e.g., mono, di, and tri-glyceryl citrates and tartrates), ethyl benzoate, ethyl acetate, ethyl carbonate, ethyl lactate, ethyl oleate, fatty acid esters of sorbitan, glyceryl monostearate, glyceride esters (e.g., mono, di, or tri-glycerides), fatty acid esters (e.g., isopropyl myristrate),
  • acetate esters e
  • Ketones which typically have from about 3 to about 30 carbons. Examples include acetone, methyl ethyl ketone, and methyl isobutyl ketone;
  • Hydrocarbons which are typically aliphatic, cycloaliphatic, or aromatic hydrocarbons having from about 4 to about 30 carbons. Examples include benzene, cyclohexane, dichloromethane, dioxolanes, hexane, n-decane, n-dodecane, n-hexane, sulfolane, tetramethylenesulfone, tetramethylenesulfoxide, toluene, dimethylsulfoxide (DMSO); and tetramethylene sulfoxide;
  • DMSO dimethylsulfoxide
  • Oils which include, for example, oils of mineral, vegetable, animal, essential, or synthetic origin. These include: mineral oils, such as aliphatic and wax-based hydrocarbons, aromatic hydrocarbons, mixed aliphatic and aromatic based hydrocarbons, and refined paraffin oil; vegetable oils, such as linseed, tung, safflower, soybean, castor, cottonseed, groundnut, rapeseed, coconut, palm, olive, corn, corn germ, sesame, persic, and peanut oil; glycerides, such as mono-, di-, and triglycerides; animal oils, such as fish, marine, sperm, cod-liver, haliver, squaiene, squalane, and shark liver oil; oleic oils; and polyoxyethylated castor oil;
  • mineral oils such as aliphatic and wax-based hydrocarbons, aromatic hydrocarbons, mixed aliphatic and aromatic based hydrocarbons, and refined paraffin oil
  • vegetable oils such as l
  • Alkyl, alkenyl, or aryl halides which include, for example, alkyl or aryl halides having from 1 to about 30 carbons and one or more halogen substituents.
  • Examples include: methylene chloride; monoethanolamine; petroleum benzin; trolamine; omega-3 polyunsaturated fatty acids (e.g., alpha-linolenic acid, eicosapentaenoic acid, docosapentaenoic acid, or docosahexaenoic acid); polyglycol ester of 12-hydroxystearic acid and polyethylene glycol (SOLUTOL HS-15, from BASF, Ludwigshafen, Germany); polyoxyethylene glycerol; sodium laurate; sodium oleate; and sorbitan monooleate.
  • Solvents useful in the invention include, but are not limited to, those known to stabilize present compounds or pharmaceutically acceptable salts thereof. These can include, for example, oils rich in triglycerides, such as safflower oil, soybean oil, and mixtures thereof; and alkyleneoxy-modified fatty acid esters, such as polyoxyl 40 hydrogenated castor oil and polyoxyethylated castor oils (e.g., CREMOPHOR EL solution or CREMOPHOR RH 40 solution).
  • oils rich in triglycerides such as safflower oil, soybean oil, and mixtures thereof
  • alkyleneoxy-modified fatty acid esters such as polyoxyl 40 hydrogenated castor oil and polyoxyethylated castor oils (e.g., CREMOPHOR EL solution or CREMOPHOR RH 40 solution).
  • triglycerides include INTRALI PID emulsified soybean oil (Kabi- Pharmacia Inc., Sweden), NUTRALIPID emulsion (McGaw, Irvine, California), LI POSYN II 20% emulsion (a 20% fat emulsion solution containing 100 mg safflower oil, 100 mg soybean oil, 12 mg egg phosphatides, and 25 mg glycerin per ml of solution; Abbott Laboratories, Chicago, IL), LIPOSYN III 2% emulsion (a 2% fat emulsion solution containing 100 mg safflower oil, 100 mg soybean oil, 12 mg egg phosphatides, and 25 mg glycerin per ml of solution; Abbott Laboratories, Chicago, IL), natural or synthetic glycerol derivatives containing the docosahexaenoyi group at levels of from about 25 to about 100% (by weight based on the total fatty acid content) (DHASCO from Martek Biosciences Corp., Columbia, MD
  • compositions of this invention for various purposes generally known in the pharmaceutical industry. These components tend to impart properties that, for example, enhance retention of the present compounds or salt thereof at the site of administration, protect the stability of the composition, control the pH, and facilitate processing of the compound or salt thereof into pharmaceutical formulations, and the like.
  • cryoprotective agents include cryoprotective agents; agents for preventing reprecipitation of the compound or salt surface; active, wetting, or emulsifying agents (e.g., lecithin, polysorbate-80, TWEEN 80, pluronic 60, and polyoxyethylene stearate); preservatives (e.g., ethyl-p-hydroxybenzoate); microbial preservatives (e.g., benzyl alcohol, phenol, m-cresol, chlorobutanol, sorbic acid, thimerosal, and paraben); agents for adjusting pH or buffering agents (e.g., acids, bases, sodium acetate, sorbitan monolaurate, etc.); agents for adjusting osmolarity (e.g., glycerin); thickeners (e.g., aluminum monostearate, stearic acid, cetyl alcohol, stearyl alcohol, guar gum, methyl cellulose, hydroxypropyl
  • Formulations for parenteral administration can be prepared from one or more sterile powders and/or granules having a compound or salt of this invention and one or more of the carriers or diluents mentioned for use in the formulations for oral administration.
  • the powder or granule typically is added to an appropriate volume of a solvent (typically while agitating (e.g., stirring) the solvent) that is capable of dissolving the powder or granule.
  • a solvent typically include, for example, water, polyethylene glycol, propylene glycol, ethanol, corn oil, cottonseed oil, peanut oil, sesame oil, benzyl alcohol, sodium chloride, and/or various buffers.
  • Emulsions for parenteral administration can be prepared by, for example, dissolving a compound or salt of this invention in any pharmaceutically acceptable solvent capable of dissolving the compound to form a solution; and adding an appropriate volume of a carrier to the solution while stirring to form the emulsion.
  • Solutions for parenteral administration can be prepared by, for example, dissolving a compound or salt of this invention in any pharmaceutically acceptable solvent capable of dissolving the compound to form a solution; and adding an appropriate volume of a carrier to the solution while stirring to form the solution.
  • Suppositories for rectal administration can be prepared by, for example, mixing the drug with a suitable nonirritating excipient that is solid at ordinary temperatures, but liquid at the rectal temperature and will therefore melt in the rectum to release the drug.
  • suitable excipients include, for example, cocoa butter; synthetic mono-, di-, or triglycerides; fatty acids; and/or polyethylene glycols.
  • Binding agents include, but are not limited to, corn starch, potato starch, or other starches, gelatin, natural and synthetic gums such as acacia, sodium alginate, alginic acid, other alginates, powdered tragacanth, guar gum, cellulose and its derivatives (e.g., ethyl cellulose, cellulose acetate, carboxymethyl cellulose calcium, sodium carboxymethyl cellulose), polyvinyl pyrrolidone, methyl cellulose, pre-gelatinized starch, hydroxypropyl methyl cellulose, (e.g., Nos. 2208, 2906, 2910), microcrystalline cellulose, and mixtures thereof.
  • natural and synthetic gums such as acacia, sodium alginate, alginic acid, other alginates, powdered tragacanth, guar gum, cellulose and its derivatives (e.g., ethyl cellulose, cellulose acetate, carboxymethyl cellulose calcium, sodium carboxymethyl
  • Suitable forms of microcrystalline cellulose include, for example, the materials sold as AVICEL-PH-101 , AVICEL- PH-103 and AVICEL-PH-105 (available from FMC Corporation, American Viscose Division, Avicel Sales, Marcus Hook, Pennsylvania, USA).
  • An exemplary suitable binder is a mixture of microcrystalline cellulose and sodium carboxymethyl cellulose sold as AVICEL RC-581 by FMC Corporation.
  • Fillers include, but are not limited to, talc, calcium carbonate (e.g., granules or powder), lactose, microcrystalline cellulose, powdered cellulose, dextrates, kaolin, mannitol, silicic acid, sorbitol, starch, pre-gelatinized starch, and mixtures thereof.
  • Lubricants include, but are not limited to, calcium stearate, magnesium stearate, mineral oil, electromagnetic radiation mineral oil, glycerin, sorbitol, mannitol, polyethylene glycol, other glycols, stearic acid, sodium lauryl sulfate, talc, hydrogenated vegetable oil (e.g., peanut oil, cottonseed oil, sunflower oil, sesame oil, olive oil, corn oil, and soybean oil), zinc stearate, ethyl oleate, ethyl laurate, agar, and mixtures thereof.
  • Additional lubricants include, for example, a syloid silica gel (AEROSIL 200, manufactured by W.R. Grace Co.
  • Disintegrants include, but are not limited to, agar-agar, alginic acid, calcium carbonate, microcrystalline cellulose, croscarmellose sodium, crospovidone, polacrilin potassium, sodium starch glycolate, potato or tapioca starch, other starches, pre-gelatinized starch, other starches, clays, other algins, other celluloses, gums, and mixtures thereof.
  • Tablets or capsules can optionally be coated by methods well known in the art. If binders and/or fillers are used with a compound/bioconjugate of the invention, they are typically formulated as about 50 to about 99 weight percent of the compound/bioconjugate. In one aspect, about 0.5 to about 15 weight percent of disintegrant, and particularly about 1 to about 5 weight percent of disintegrant, can be used in combination with the compound. A lubricant can optionally be added, typically in an amount of less than about 1 weight percent of the compound/bioconjugate.
  • Liquid preparations for oral administration can take the form of solutions, syrups or suspensions. Alternatively, the liquid preparations can be presented as a dry product for constitution with water or other suitable vehicle before use.
  • Such liquid preparations can be prepared by conventional means with pharmaceutically acceptable additives such as suspending agents (e.g., sorbitol syrup, cellulose derivatives or hydrogenated edible fats); emulsifying agents (e.g., lecithin or acacia); non-aqueous vehicles (e.g., almond oil, oily esters, ethyl alcohol or fractionated vegetable oils); and/or preservatives (e.g., methyl or propyl-p- hydroxybenzoates or sorbic acid).
  • suspending agents e.g., sorbitol syrup, cellulose derivatives or hydrogenated edible fats
  • emulsifying agents e.g., lecithin or acacia
  • non-aqueous vehicles e.g., almond oil, oily esters,
  • the preparations can also contain buffer salts, flavoring, coloring, perfuming and sweetening agents as appropriate.
  • Preparations for oral administration can also be formulated to achieve controlled release of the compound/bioconjugate.
  • Oral formulations preferably contain 10% to 95% compound/bioconjugate.
  • a compound/bioconjugate of the present invention can be formulated for buccal administration in the form of tablets or lozenges formulated in a conventional manner.
  • Other methods of oral delivery of compounds/bioconjugates of the invention will be known to the skilled artisan and are within the scope of the invention.
  • Hard gelatin capsules are prepared using the following ingredients:
  • a tablet formula is prepared using the following ingredients:
  • a dry powder inhaler formulation is prepared containing the following components: TABLE F3
  • the active ingredient is mixed with the lactose and the mixture is added to a dry powder inhaling appliance.
  • the active ingredient, starch and cellulose are passed through a No. 20 mesh U.S. sieve and mixed thoroughly.
  • the solution of polyvinylpyrrolidone is mixed with the resultant powders which are then passed through a 16 mesh U.S. sieve.
  • the granules as produced are dried at 50-60 °C and passed through a 16 mesh U.S. sieve.
  • the sodium carboxymethyl starch, magnesium stearate, and talc previously passed through a No. 30 mesh U.S. sieve, are then added to the granules which, after mixing, are compressed on a tablet machine to yield tablets each weighing 150 mg.
  • Capsules each containing 80 mg of active ingredient are made as follows: TABLE F5
  • Suppositories each containing 225 mg of active ingredient, are made as follows:
  • the active ingredient is passed through a No. 60 mesh U.S. sieve and suspended in the saturated fatty acid glycerides previously melted using the minimum heat necessary. The mixture is then poured into a suppository mold of nominal 2.0 g capacity and allowed to cool.
  • the active ingredient, sucrose and xantham gum are blended, passed through a No. 10 mesh U.S. sieve, and mixed with a previously made solution of the microcrystalline cellulose and sodium carboxymethyl cellulose in water.
  • the sodium benzoate, flavor, and color are diluted with some of the water and added with stirring. Sufficient water is then added to produce the required volume.
  • Capsules each containing 150 mg of active ingredient, are made as follows:
  • kits can include a compound/bioconjugate of the present invention, optionally one or more ingredients for preparing a pharmaceutically acceptable formulation of the compound/bioconjugate, and instructions for use (e.g., administration).
  • a kit different components of a compound/bioconjugate formulation can be packaged in separate containers and admixed immediately before use.
  • Such packaging of the components separately can, if desired, be presented in a pack or dispenser device which can contain one or more unit dosage forms containing the compound/bioconjugate.
  • the pack can, for example, comprise metal or plastic foil such as a blister pack.
  • Such packaging of the components separately can also, in certain instances, permit long-term storage without losing activity of the components.
  • the different components can be packaged separately and not mixed prior to use. In various embodiments, the different components can be packaged in one combination for administration together.
  • the compounds and salts of this invention can be used in the form of a kit that is suitable for use in performing the methods described herein, packaged in a container.
  • the kit can contain the compound or compounds and, optionally, appropriate diluents, devices or device components suitable for administration and instructions for use in accordance with the methods of the invention.
  • the devices can include parenteral injection devices, such as syringes or transdermal patch or the like.
  • Device components can include cartridges for use in injection devices and the like.
  • the kit includes a first dosage form including a compound or salt of this invention and a second dosage form including another active ingredient in quantities sufficient to carry out the methods of the invention.
  • the first dosage form and the second dosage form together can include a therapeutically effective amount of the compounds for treating the targeted condition(s).
  • kits can be supplied with instructional materials. Instructions can be printed on paper or other substrate, and/or can be supplied as an electronic-readable medium, such as a floppy disc, mini-CD-ROM, CD-ROM, DVD-ROM, Zip disc, videotape, audio tape, and the like. Detailed instructions can not be physically associated with the kit; instead, a user can be directed to an Internet web site specified by the manufacturer or distributor of the kit, or supplied as electronic mail.
  • the emulsions or solutions described above for oral or parenteral administration can be packaged in IV bags, vials, or other conventional containers in concentrated form, and then diluted with a pharmaceutically acceptable liquid (e.g., saline) to form an acceptable compound concentration before use.
  • a pharmaceutically acceptable liquid e.g., saline
  • Kits can include reagents in separate containers such as, for example, sterile water or saline to be added to a lyophilized active component packaged separately.
  • sealed glass ampules can contain lyophilized superoxide dismutase mimetics and in a separate ampule, sterile water, sterile saline or sterile each of which has been packaged under a neutral non-reacting gas, such as nitrogen.
  • Ampules can consist of any suitable material, such as glass, organic polymers, such as polycarbonate, polystyrene, ceramic, metal or any other material typically employed to hold reagents.
  • suitable containers include bottles that can be fabricated from similar substances as ampules, and envelopes that can consist of foil- lined interiors, such as aluminum or an alloy.
  • Other containers include test tubes, vials, flasks, bottles, syringes, and the like.
  • Containers can have a sterile access port, such as a bottle having a stopper that can be pierced by a hypodermic injection needle.
  • Other containers can have two compartments that are separated by a readily removable membrane that upon removal permits the components to mix.
  • Removable membranes can be glass, plastic, rubber, and the like.
  • diastereomer of the compound is not specified, for example, in a formula or in a chemical name, that description is intended to include each isomers and enantiomer of the compound described individual or in any combination.
  • all isotopic variants of compounds disclosed herein are intended to be encompassed by the disclosure.
  • any one or more hydrogens in a molecule disclosed can be replaced with deuterium or tritium.
  • Isotopic variants of a molecule are generally useful as standards in assays for the molecule and in chemical and biological research related to the molecule or its use. Methods for making such isotopic variants are known in the art. Specific names of compounds are intended to be exemplary, as it is known that one of ordinary skill in the art can name the same compounds differently.
  • ranges specifically include the values provided as endpoint values of the range.
  • ranges specifically include all the integer values of the range. For example, a range of 1 to 100 specifically includes the end point values of 1 and 100. It will be understood that any subranges or individual values in a range or subrange that are included in the description herein can be excluded from the claims herein.

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Abstract

The invention relates generally to optical agents for biomedical applications including imaging, visualization, phototherapy and diagnostic monitoring of cells and tissue expressing MMPs and/or tissues associated with the expression of MMPs. In some embodiments, for example, optical agents of the present invention selectively bind to, or otherwise preferentially associate with, a target tissue expressing one or more MMPs, and function to couple electromagnetic radiation into and/or out of the target tissue, for example via optical absorption, fluorescence, scattering and/or optoacoustic processes.

Description

OPTICAL AGENTS FOR IMAGING AND VISUALIZATION OF
MATRIX METALLOPROTEINASE ENZYMES
CROSS-REFERENCE TO RELATED APPLICATIONS
[001] This application claims the benefit of priority of U.S. Provisional Patent Application No. 61/533,409, filed September 12, 201 1 , which is hereby incorporated by reference in its entirety.
BACKGROUND
[002] Matrix metalloproteinases are a class of zinc-dependent endopeptidases having functionality for degradation and remodeling of the extracellular matrix and subendothelial basement membrane. This family of more than 20 enzymes is further classified in several main groups, including gelatinases, interstial collagenases, stromelysins, and membrane-type matrix metalloproteinases, and is typically referenced using the abbreviation "MMP," optionally followed by a number providing differentiation between specific enzymes of the family (e.g., MMP-1 , MMP-2, etc.). Collectively, MMPs are capable of degrading all components of the extracellular matrix, including fibrallar and non-fibrallar collagens, fibronectin, laminin, and basement membrane glycoproteins. The biological activity of MMPs is regulated on a number of levels. MMP genes are transcriptionally responsive to a variety of oncogenes, growth factors, cytokins and hormones. In addition, MMP activity is regulated by the combination of secretion, enzyme mediated activation and inhibition via naturally occurring MMP inhibitors such as a2- macroglobulin and a variety of tissue inhibitors of metalloproteinases (TIMPs).
[003] MPPs are believed to play a role in a number of physiological and pathological processes involved in the onset and progression of certain diseases characterized by breakdown of connective tissue and/or extracellular matrix. Pathological conditions for which MMPs are thought to be implicated include: rheumatoid arthritis; osteoarthritis; septic arthritis, corneal, epidermal and gastric ulceration; tumor metastasis, invasion and angiogenesis;
coronary thrombosis; Alzheimer's disease and bone disease. In particular, there is considerable evidence demonstrating a close association between expression of certain MMPs and the proliferation, invasive behavior and metastatic potential of certain types of tumor. [See, e.g., Heath E.I., Grochow L.B.: Clinical potential of matrix metalloprotease inhibitors in cancer therapy. Drugs 2000, 59, 1043-1055; Hidalgo M., Eckhardt G.: Development of matrix metalloproteinase inhibitors in cancer therapy. J Natl Cancer Inst 2001 , 93, 178-193; and Stamenkovic I.: Matrix metalloproteinases in tumor invasions and metastasis. Emin. Cancer Biol. 2000, 10, 415-433]. MMP-2 and MMP-9, for example, are frequently coexpressed in a number of human cancers, such as colorectal carcinomas, breast cancer and nonsmall cell lung cancers. This observation has lead to an understanding that MMPs play a critical role in establishing and maintaining a macroenvironment that facilitates growth and angiogenesis of tumors at primary and metastatic sites. [See, e.g., Zucker S., Lysik R.M., Zarrabi M.H., Moll U.: Mr 92,00 Type IV collagenase is increased in plasma of patients with colon and breast cancer. Cancer Res 53:140-146, 1993; and Brown P.D., Bloxidge R.E., Stuart NSA, Gatter K.C., Carmichael J.: Association between expression of activated 72-kilodalton gelatinase and tumor spread in non-small cell lung carcinoma. J Natl Cancer Inst 85:574-578, 1993.] In addition, MMPs are believed to be involved with biological processes associated with inflammation, cardiovascular disease and acute and chronic neuropathic pain states. Elevated serum levels of MMP-3, for example, have been observed in rheumatoid arthritis patients and have also been shown to correlate with systemic markers of inflammation under some conditions. [See, e.g., Yoshihara Y., Obata K., Fujimoto N., Yamashita K., Hayakawa T., Shimmei M.: Increased levels of stromelysin-1 and tissue inhibitor of metalloproteinases-1 in sera from patients with rheumatoid arthritis. Arthritis Rheum 38:969-975, 1995; Hembry R.M., Bagga M.R., Reynolds J. J., Hamblen D.L.: Immunolocalisation studies on six matrix metalloproteinases and their inhibitors, TIMP-1 and TIMP-2, in synovial from patients with osteo- and rheumatoid arthritis. Ann Rheum Dis 54:25-32, 1995; and Manicourt D.H., Fujimoto N., Obata K., Thonar E.J.:
Serum levels of collagenase, stromelysin-1 , and TIMP-1 . Age- and sex-related differences in normal subjects and relationship to the extent of joint involvement and serum levels of antigenic keratin sulfate in patients with osteoarthritis. Arthritis Rheum 37:1774-1783, 1994.]
[004] Given their role in tumor progression and metastasis processes, there is considerable interest in MMPs as biological targets for development of therapies for the treatment of cancer. Therapeutic approaches being pursued include the development of psuedopeptides having structural components similar to MMP substrates which function as competitive, reversible inhibitors and the development of nonpeptide, low molecular weight MMP inhibitors. MMPs inhibitors in development and in various stages of clinical trials include broad spectrum inhibitors capable of simultaneously blocking multiple MMPs. The broad spectrum MMP inhibitor marmastat, for example, has been reported as effective for the treatment of gastric cancer and pancreatic cancer. Prinomastat is another broad spectrum MMP inhibitor in advanced clinical trials for the treatment of cancer. Selective MMP inhibitors are also in development that are capable of blocking activity of a selected MMP or group of MMPs associated with specific pathological condition. A motivation for development of selective MMP inhibitors is enhancing efficacy for certain therapeutic applications and reducing unwanted side effects associated with blocking the activity of certain MMPs.
[005] A large number of compounds and composition classes have been proposed and/or evaluated for the inhibition of MMPs. A class of thiol group-containing amide or peptidyl amide- based compounds alleged to exhibit activity for MMP inhibition are disclosed in U.S. Patent No. 4,595,700 and PCT patent publications WO 95/012389 and WO 96/01 1209. A number of groups have developed hydroxamate-containing compounds for MMP inhibition. U.S. Patent No. 6,492,367, for example, discloses a class of sulfamato hydroxamic acid compounds alleged to exhibit activity for MMP inhibition. In addition, Becker et al. describe a series of a-amino-β- sulfone hydroxamates, a-alkyl-a-amino-p-sulfone hydroxamates, and a-piperidine-p-sulfone hydroxamates as anti-tumor and/or anti-angiogenesis agents that are alleged to exhibit activity for selective MMP inhibition. [See, Becker et al.: Synthesis and Structure - Activity
Relationships of β- and a-Piperidine Sulfone Hydroxamic Acid Matrix Metalloproteinase
Inhibitors with Oral Antitumor Efficacy. J Med Chem, 2005, 48(21 ), 6713-6730; Becker et al..: a-Amino-3-sulfone hydroxamates as potent MMP-13 inhibitors that spare MMP-1. Bioorg Med Chem Lett 2001 , 1 1 , 2719-2722; and Becker et al..: a-Alkyl-a-amino-3-sulfone hydroxamates as potent MMP inhibitors that spare MMP-1 . Bioorg Med Chem Lett 2001 , 1 1 , 2723-2725].
Further, Aranapakam et al. describe a series of a-sulfone hydroxamates developed in the context of treating osteoarthritis that are alleged to exhibit activity for MMP inhibition.
[Aranapakam et al., Synthesis and structure-activity relationship of a-sulphonyl-hydroxamic acids as novel, orally active matrix metalloproteinase inhibitors for the treatment of osteoarthritis; J. Med. Chem., 2003, 46, 2361-2375; and Aranapakam et al., Synthesis and structure-activity relationship of N-substituted 4-arylsulfonylpiperidine-4-hydroamic acids as novel, orally active matrix metalloproteinase inhibitors for the treatment of osteoarthritis; J. Med. Chem., 2003, 46, 2376-2396].
[006] In addition to their role as therapeutic agents, MMPs are also of potential interest as diagnostic agents. For example, a number of studies demonstrate the potential for various classes of MMPs to provide biomarkers for identifying the onset and the prognosis of certain cancers. Expression of stromelysin-3 has been observed in invasive breast carcinomas and is not observed in normal breast tissue, thereby supporting a potential diagnostic application of this class of MMPs. [See, e.g., 30. Ahmed A., Hanby A., Dublin E., et al: Stromelysin-3: An independent prognostic factor for relapse-free survival in node-positive breast cancer and demonstration of novel breast carcinoma cell expression. Am J Pathol 152:721 -728, 1998; and Tetu B., Brisson J., Lapointe H., et al: Prognostic significance of stromelysin 3, gelatinase A, and urokinase expression in breast cancer. Hum Pathol 29:979-985, 1998.]. In addition, studies show elevated levels of serum gelatinase A in patients with prostate cancer relative to those without the disease. [See, e.g., Gohji K., Fujimoto N., Hara I., et al: Serum matrix metalloproteinase-2 and its density in men with prostate cancer as a new predictor of disease extension. Int J Cancer 79:96-101 , 1998.]. In addition, the expression of matrilysin (MMP 7) has also been suggested to be of prognostic value in evaluation of colon and esophageal cancers. [See, e.g., Ichikawa, Y. et al., Detection of regional lymph node metastases in colon cancer by using RT-PCT for matrix metalloproteinase-7, matrilysin; Clin. Exp. Metastasis, 16:3- 8, 1998; and Yamamoto, H. et al., Association of matrilysin expression with recurrence and poor prognosis in human esophageal squamous cell carcinoma,; Cancer Res. 59:3313-3316, 1999.]
[007] While MMPs are a well recognized target for development of therapeutic and diagnostic agents, their potential for biomedical imaging and visualization remains much less explored. Development of optical agents for imaging expression of MMPs in certain tumors and lesions, for example, has potential application for diagnosis, staging and monitoring of cancer and for the evaluation of tissue degradation associated with pathological conditions including heart disease and arthritis. In addition, optical agents capable of selective delivery to cells and tissue expressing MMPs also has potential for visually differentiating the tissue of a tumor or lesion from normal tissue during a surgical procedure, such as a biopsy or therapeutic tissue removal procedure.
[008] In addition to the ability to selectively associate with tissue expressing MMPs under in vivo conditions, the effectiveness of optical agents for optical imaging and visualization applications involving MMP targets depends on an important combination of optical,
physiochemical and pharmacokinetic properties. First, optical agents preferably have large extinction coefficients for visible and near infrared (NIR) electromagnetic radiation having wavelengths from about 350 nm to about 900 nm, and for fluorescent optical agents optionally have fluorescence quantum yields and Stokes shifts useful for fluorescence imaging and detection. Second, optical agents for these applications preferably exhibit low systemic toxicity, low mutagenicity, and rapid clearance from the blood stream. Third, optical agents for these applications are preferably compatible with effective formulation and administration to a target tissue expressing MMPs, for example by exhibiting a reasonable solubility in aqueous solution, a low tendency for aggregation in solution and/or compatibility with pharmaceutical delivery systems, such as liposomes or micelle delivery agents.
[009] As will be generally recognized from the foregoing, a need currently exists for optical agents for imaging and visualization for diagnosis, evaluation and treatment of disorders associated with expression of MMPs. Specifically, optical agents for imaging and visualization are needed exhibiting specificity for important target tissues expressing MMPs, such as tumors and other lesions. In addition, optical agents are needed having optical, physiochemical and pharmacokinetic properties providing for useful administration, delivery and excitation with electromagnetic radiation.
SUMMARY
[010] The invention relates generally to optical agents for biomedical applications including imaging, visualization, phototherapy and diagnostic monitoring of cells and tissues expressing MMPs and/or tissues associated with the expression of MMPs. In some embodiments, for example, optical agents of the present invention selectively bind to, or otherwise preferentially associate with, a target tissue expressing one or more MMPs, and function to couple
electromagnetic radiation into and/or out of the target tissue, for example via optical absorption, fluorescence, scattering and/or optoacoustic processes. The present optical agents enable a versatile diagnostic platform useful for in vivo, in vitro and ex vivo diagnostic monitoring, visualization and imaging applications, such as, but not limited to, tomographic, photoacoustic and/or sonofluorescent imaging; monitoring and evaluating organ functioning; anatomical visualization; coronary angiography; and fluorescence endoscopy. In some embodiments, the invention provides tumor-specific optical agents for detection, diagnosis and/or imaging of cancer. In some embodiments, optical agents of the present invention provide tandem imaging and therapeutic functionality, including phototherapy, for example by simultaneously functioning as a MMP inhibitor, an optical probe and/or a phototherapeutic agent, such as a Type 1 or Type 2 phototherapy agent.
[011] The invention provides optical agents, including compositions, preparations and formulations, and methods of using and making optical agents for diagnostic and clinical applications. Optical agents of the invention include compounds having a sulfone hydroxamic acid or hydroxamate backbone component providing molecular recognition and/or targeting functionality for one or more MMPs and/or tissues expressing or associated with expression of MMPs and an optical dye component providing preselected optical properties, such as absorption and/or emission in the visible and/or near infrared region of the electromagnetic spectrum (e.g., 350 nm - 1300 nm or preferably for some applications 400 nm to 900 nm). In an embodiment, for example, an optical agent of the invention comprises a sulfone hydroxamic acid or hydroxamate derivative including a pyrazine, cyanine, indocyanine, azulene or azaazulene component having ring substituents providing large absorption in the visible and/or near infrared region of the electromagnetic spectrum, and optionally a fluorescence quantum yield and Stokes shift useful for optical excitation and detection in biomedical procedures.
Pyrazine-containing, cyanine-containing, indocyanine-containing, azulene-containing and/or azaazulene-containing sulfone hydroxamic acid or hydroxamate derivatives of some
embodiments of the present invention provide exogenous optical agents for biomedical and bioanalytical applications including imaging, visualization, diagnostic monitoring, and therapy.
[012] In an aspect, the invention provides compounds being of formula (FX1 ) or (FX2):
); or
Figure imgf000007_0001
(FX2); or a pharmaceutically acceptable salt or ester thereof; wherein: each of L1, L2, and LJ, if present, is independently CrC10 alkylene, C3-C10 cycloalkylene, C2-Ci0 alkenylene, C3-C10 cycloalkenylene, C2-Ci0 alkynylene, ethenylene, ethynylene, phenylene, -CO-, -0-, -S-, -(CH2CH20)a-, -CO(CH2CH20)a- -(CHOH)b- or - CO(CHOH)b-; each of W1 , W2, and W3 is independently a single bond, -(CH2)n- -(HCCH)n- - (CH2CH20)a- -0-, -S-, -SO-, -S02-, -SO3-, -OS02-, -NR11-, -CO-, -COO-, -OCO-, - OCOO-, -CONR12-, -NR13CO- -OCONR14-, -NR15COO- -NR16CONR17-, -NR18CSNR19-, - (CH2)mO(CH2)n-, -(CH2)mS(CH2)n-,-(CH2)mSO(CH2)n, -(CH2)m S02(CH2)n-, -(CH2)mS03(CH2)n-, -(CH2)mOS02(CH2)n-, -(CH2)mNR20(CH2)n-, -(CH2)mCO(CH2)n-, -(CH2)mCOO(CH2)n-, - (CH2)mOCO(CH2)n-, -(CH2)mOCOO(CH2)n-, -(CH2)mCONR21 (CH2)n-,-(CH2)mNR22CO(CH2)n-, - (CH2)mOCONR23(CH2)n- -(CH2)mNR24COO(CH2)n- -(CH2)mNR25CONR26(CH2)n- - (CH2)mNR27CSNR28(CH2)n- -(CH2)mO(CH2)nNR29CO(CH2)n- - (CH2)mCO(CH2)n(CH2OCH2)a(CH2)nNR30(CH2)nNR31 CO- -CO(CH2)nNR32CO- or - CONR33(CH2)nNR34-;
each ring Z is independently
Figure imgf000008_0001
each U is independently -0-, -S-, -CR8R9- or -NR10-; each Y is independently -N- or -CR2-; each A is independently a group corresponding to an optical dye; each of R1 to R5 and R8 to R10 is independently hydrogen, -OCF3, CrC6 alkyl, C3-C6 cycloalkyl, C5-C10 aryl, C5-C10 hetroaryl, Ci-C6 acyl, C2-C6 alkenyl, C2-C6 alkynyl, C5-C10 alkylaryl, halo, halomethyl, dihalomethyl, trihalomethyl, -CN, -C02R60, -CONR61 R62, -COR63, -N02, -SOR64, -OSR65 , -S02R66, - S02OR67, -S02NR68R69, -PO3R70R71 , -OR72, -SR73, -NR74R75, -NR76COR77, - CH2(CHOH)cR78,-(CH2CH20)cR79, PS1 or PS2; each of R11 to R34 and R60 to R79 is independently hydrogen, CrC6 alkyl, C3-C6 cycloalkyl, C5-C10 aryl or C5-C10 heteroaryl; each PS1 is
independently an azide, azo, diazo, oxaza, diaza, dithia, thioxa, or dioxa group; each PS2 is independently a group corresponding to a porphyrin, benzoporphyrin, phthalocyanine, phenothiazine, chlorin, bacteriochlorin, phthalocyanine, porphyrin, purpurin, merocyanine, pheophorbides, psoralen, aminolevulinic acid (ALA), hematoporphyrin derivative, porphycenes, or porphacyanine; each c is independently 0 or an integer selected from the range of 1 to 100; each of a and b is independently an integer selected from the range of 1 to 100; each of m and n is independently 0 or an integer selected from the range of 1 to 10; and each of q, r and x is independently 0 or 1. Compounds of the invention, for example compounds having formula (FX1 ) - (FX113), are useful as optical agents, for example, in an optical imaging, diagnostic, and/or therapeutic biomedical procedure.
[013] In an embodiment, for example, the invention provides compounds being of the formula (FX3) or (FX4):
Figure imgf000009_0001
(FX4); or a pharmaceutically acceptable salt or ester thereof; wherein L1-L2, W1-W2, ring Z, A, R1-R5, q, and x are as set forth in the description of formulas (FX1 ) and (FX2).
[014] The term "optical dye" refers to a functional group of a compound that absorbs, emits, and/or scatters electromagnetic radiation in the visible and/or infrared regions of the
electromagnetic spectrum, for example, electromagnetic radiation having wavelengths selected over the range of 350 - 1300 nanometers, and optionally for some applications over the range of 400 nm to 900 nm. Optical dyes of some compounds of the invention function as a chromophore, fluorophore and/or functional group exhibiting photoacoustic properties, for example, when provided in an in vivo environment. Optical dyes of some compounds of the invention enable the compound to undergo emission processes upon absorption of
electromagnetic radiation, for example via fluorescence and/or phosphorescence pathways. Optical dyes of some compounds of the invention enable the compound to undergo
fluorescence of visible or infrared electromagnetic radiation, for example, via emission of electromagnetic radiation having wavelengths selected over the range of 350 - 1300 nanometers, and optionally for some applications over the range of 400 nm to 900 nm. Optical dyes of some compounds of the invention enable the compound to undergo fluorescence of visible or infrared electromagnetic radiation, for example, by exhibiting a fluorescence quantum yield in an in vivo environment that is greater than or equal to 0.0001 , optionally for some embodiments greater than or equal to 0.001 , optionally for some embodiments greater than or equal to 0.01 , and optionally for some embodiments greater than or equal to 0.1. Optical dyes of some compounds of the invention enable the compound to undergo fluorescence of visible or infrared electromagnetic radiation, for example, by exhibiting a Stokes shift in an in vivo environment greater than or equal to 5 nanometers, optionally for some embodiments greater than or equal to 10 nanometers and optionally for some embodiments greater than or equal to 20 nanometers. Optical dyes in some compounds are directly or indirectly covalently linked to the sulfone hydroxamic acid or hydroxamate backbone of the compound, for example, by linking groups and/or spacing groups that do not interfere with the molecular recognition and/or targeting functionality of the hydroxamic acid or hydroxamate backbone with respect to MMPs and/or tissues associate with MMPs.
[015] In an embodiment, the invention provides compounds having any of formula (FX1 ) - (FX27), wherein A is a group corresponding to a pyrazine, a cyanine, an indocyanine, phthalocyanine, a rhodamine, a thiazole, a phenylxanthene, a phenothiazine, a
phenoselenazine, a squaraine, a dipyrrolo pyrimidone, an anthraquinone, a tetracene, a quinoline, an acridine, an acridone, a phenanthridine, an azo dye, a phenoxazine, an azulene, an azaazulene, a triphenyl methane dye, an indole, a benzoindole, an indocarbocyanine, a Nile Red dye, a thionin dye, an isosulfan blue dye, or a benzoindocarbocyanine, including for example, derivatives thereof having one or more electron withdrawing groups, electron donating groups and/or photosensitizer groups. In an embodiment, for example, a compound having any of formula (FX1 ) - (FX27), is provided wherein A is a group corresponding to a pyrazine, a cyanine, azulene, azaazulene or an indocyanine, including for example, derivatives thereof having one or more electron withdrawing groups, electron donating groups and/or
photosensitizer groups.
[016] As used throughout the present description, reference to embodiments wherein q, x and/or r is equal to 0 refers to compounds where L1, L2 or L3, respectively, is not present and reference to embodiments wherein q, x and/or r is equal to 1 refers to compounds where L1, L2 or L3 , respectively, is present. For example, W2 is directly linked to ring Z when x is equal to 0 and W3 is directly linked to ring Z when r is equal to 0. For example, W1 is directly linked to an adjacent piperidine group when q is equal to 0. Embodiments wherein W2 is a single bond and x is equal to 0 refer to compositions having a phenyl group or piperidine group (optionally substituted) directly bonded to ring Z and embodiments wherein W3 is a single bond and r is equal to 0 refer to formula having an optical dye (optionally substituted) directed bonded to ring Z. Embodiments wherein W1 is a single bond and q is equal to 0 refer to compositions having an optical dye (optionally substituted) directed bonded to an adjacent piperidine group. As used throughout the present description, the expression "a group corresponding to" an indicated species expressly includes a radical of the species or group, such as an aromatic radical or heterocyclic aromatic radical of the species or group of species provided in a covalently bonded configuration, optionally with one or more substituents, including but not limited to electron donating groups, electron withdrawing groups, photosensitizers and/or targeting ligands.
[017] The sulfone hydroxamic acid backbone of some compounds having formula (FX1 ) or (FX2) provides selective binding with MMPs and the optical dye component provides useful optical functionality. Some of the present optical agents having formula (FX1 ), for example, have the optical dye component provided in a solvent exposed portion of the MMP binding pocket of the optical agent. Alternatively, some optical agents having formula (FX2), for example, have the optical dye component provided in the SV MMP binding pocket of the optical agent. Optical agents of the present invention providing the optical dye component in the solvent exposed portion of the MMP binding pocket or in the SV MMP binding pocket are particularly useful for some applications, as binding of the optical agent to a MMP may generate an observable change in the optical properties of the optical agent, such as an observable change in the excitation or emission wavelengths, fluorescence quantum yield or Stokes shift, which may be useful for enhancing optical detection, imaging or monitoring cells and/or tissue expressing MMPs or associate with MMP expression.
[018] In an aspect, the invention provides compounds being of the formula (FX5), (FX6), (FX7) or (FX8):
Figure imgf000011_0001
Figure imgf000011_0002
[019] "u" (FX8); or a pharmaceutically acceptable salt or ester thereof; wherein: L1 to L3, W1 to W3, A, R1 to R5, q, r, x and U are as set forth in the description of formula (FX1 ) and (FX2).
[020] In an aspect, the invention provides compounds being of the formula (FX9), (FX10),
Figure imgf000012_0001
(FX12); or a pharmaceutically acceptable salt or ester thereof; wherein: L1 to L3, W1 to W3, A, R1 to R5, q, r, x and U are as set forth in the description of formula (FX1 ) and (FX2).
[021] In an aspect, the invention provides compounds being of the formula (FX13) or (FX14):
Figure imgf000013_0001
eutically acceptable salt or ester thereof; wherein: B is -O- or -S- and wherein Ring Z, L1, W1, A, R1 to R5, q, and U are as set forth in the description of formula (FX1 ) and (FX2). In an aspect, the invention provides compounds being of the formula (FX15) or (FX16):
Figure imgf000013_0002
acceptable salt or ester thereof; wherein: Ring Z, L1, W1, A, R1 to R5, q, and U are as set forth in the description of formula (FX1 ) and (FX2). In an aspect, the invention provides compounds being of the formula (FX17), (FX18) or (FX19):
Figure imgf000014_0001
(FX19); or a pharmaceutically acceptable salt or ester thereof: wherein Ring Z, L3, W3, A, R10, r, and U are as set forth in the description of formula (FX1 ) and (FX2).
[022] In an aspect, the invention provides compounds being of the formula (FX20), (FX21 ), (FX22) or (FX23):
Figure imgf000014_0002
Figure imgf000015_0001
or a pharmaceutically acceptable salt or ester thereof; wherein: B is -O- or -S-; and wherein Ring Z, A, n and m are as set forth in the description of formula (FX1 ) and (FX2).
[023] In an aspect, the invention provides compounds being of the formula (FX24), (FX25),
Figure imgf000015_0002
Figure imgf000016_0001
(FX27); or a pharmaceutically acceptable salt or ester thereof; wherein: B is -O- or -S-;wherein ring Z, R1 to R5, n, m, a, and A are as set forth in the description of formula (FX1 ) and (FX2).
[024] In an embodiment, the invention provides compounds having any one of formula (FX1 ) - (FX27), wherein A is a group corresponding to a pyrazine, for example a pyrazine providing a chromophore and/or fluorophore that absorbs electromagnetic radiation having wavelengths in the visible and/or infrared regions of the electromagnetic spectrum (e.g., 350 nm to 1300 nm, or optionally 400 nm to 900 nm). Embodiments of the invention include, for example, compounds having any one of formula (FX1 ) - (FX27), wherein A is a group corresponding to a pyrazine having one or more electron donating groups, electron withdrawing groups and/or
photosensitizer groups as substituents.
[025] In an aspect, the invention provides compounds being of the formula (FX28) or (FX29):
Figure imgf000016_0002
or a pharmaceutically acceptable salt or ester thereof; wherein each of X1, X2 and X3 is independently hydrogen, - OCF3, C C6 alkyl, C3-C6 cycloalkyl, C5-C10 aryl, C5-C10 heteroaryl, C C6 acyl, C2-C6 alkenyl, C2- C6 alkynyl, C5-C10 alkylaryl, halo, halomethyl, dihalomethyl, trihalomethyl, -C02R4°, -SOR41 , - OSR42, -S02OR43, -CH2(CH2OCH2)cCH2OH, -P03R44R45, -OR46, -SR47, -NR48R49, - NR50COR51 , -CN, -CONR52R53, -COR54, -N02, -S02R55, -S02NR56R57, PS1 or PS2; and each of R40 to R57 is independently hydrogen, C C6 alkyl, C3-C6 cycloalkyi, C5-C10 aryl, C5-C10 heteroaryl, or pyrrolyl; wherein U, Ring Z, L1 to L3, W1 to W3, R1 to R5, x, r, and q are as set forth in the description of formula (FX1 ) and (FX2).
[026] In an aspect, the invention provides compounds being of the formula (FX30), (FX31 ),
Figure imgf000017_0001
Figure imgf000018_0001
(FX33); or a pharmaceutically acceptable salt or ester thereof; wherein: U, L1 to L3, W1 to W3, R1 to R5, X1 to X3, x, r, and q are as set forth in the description of formula (FX1 ), (FX2), (FX28) and (FX29).
[027] In an aspect, the invention provides compounds being of the formula (FX34), (FX35), (FX36) or (FX37):
Figure imgf000018_0002
Figure imgf000019_0001
(FX37); or a pharmaceutically acceptable salt or ester thereof; wherein: U, L1 to L3, W1 to W3, R1 to R5, X1 to X3, x, r, and q are as set forth in the description of formula (FX1 ), (FX2), (FX28) and (FX29).
[028] In an aspect, the invention provides compounds being of the formula (FX38) or (FX39):
(FX38); or
Figure imgf000019_0002
(FX39); or a pharmaceutically acceptable salt or ester thereof; wherein: B is -O- or -S-; wherein Ring Z, U, L1, W1, R1 to R5, X1 to X3, and q are as set forth in the description of formula (FX1 ), (FX2), (FX28) and (FX29).
[029] In an aspect, the invention provides compounds being of the formula (FX40) or (FX41 ):
Figure imgf000020_0001
(FX41 ); or a pharmaceutically acceptable salt or ester thereof; wherein: Ring Z, U, L1, W1, R1 to R5, X1 to X3, and q are as set forth in the description of formula (FX1 ), (FX2), (FX28) and (FX29).
[030] In an aspect, the invention provides compounds being of the formula (FX42), (FX43) or (FX44):
Figure imgf000020_0002
Figure imgf000021_0001
(FX44); or a pharmaceutically acceptable salt or ester thereof; wherein: Ring Z, U, L3, W3, R1 to R5, X1 to X3, R10 and r are as set forth in the description of formula (FX1 ), (FX2), (FX28) and (FX29).
[031] In an aspect, the invention provides compounds being of the formula (FX45), (FX46), (FX47) or (FX48):
Figure imgf000021_0002
a pharmaceutically acceptable salt or ester thereof; wherein: B is -O- or -S-; wherein Ring Z, U, m, n, and X1 to X3 are as set forth in the description of formula (FX1 ), (FX2), (FX28) and (FX29).
[032] In an aspect, the invention provides compounds being of the formula (FX49), (FX50), (FX51 ) or (FX52):
Figure imgf000022_0001
(FX52); or a pharmaceutically acceptable salt or ester thereof; wherein: m, n, X1 to X3, R10, R48, R49, and R54 are as set forth in the description of formula (FX1 ), (FX2), (FX28) and (FX29).
[033] In an aspect, the invention provides compounds being of the formula (FX53), (FX54), (FX55) or (FX56):
Figure imgf000023_0001
(FX53);
Figure imgf000023_0002
(FX54);
Figure imgf000023_0003
(FX55); or
Figure imgf000024_0001
pharmaceutically acceptable salt or ester thereof; wherein: B is -O- or -S-; wherein ring Z, a, m, n, X1 to X3, R1 to R5 are as set forth in the description of formula (FX1), (FX2), (FX28) and (FX29).
[034] In an aspect, the invention provides compounds being of the formula (FX57), (FX58), or
Figure imgf000024_0002
Figure imgf000025_0001
48 ->49 ->54 pharmaceutically acceptable salt or ester thereof; wherein: a, m, n, R , R , R , and R are as set forth in the description of formula (FX1 ), (FX2), (FX28) and (FX29).
[035] In an aspect, the invention provides compounds being of the formula (FX60), (FX61 ), or
Figure imgf000025_0002
R48 R49 (FX62); or a pharmaceutically acceptable salt or ester thereof; wherein: R48 and R49 are as set forth in the description of formula (FX1 ), (FX2), (FX28) and (FX29).
[036] In an embodiment, the invention provides compounds having any one of formula (FX1 ) - (FX27), wherein A is a group corresponding to an indocyanine, for example an indocyanine providing a chromophore and/or fluorophore that absorbs electromagnetic radiation having wavelengths in the visible and/or infrared regions of the electromagnetic spectrum (e.g., 350 nm to 1300 nm, or optionally 400 nm to 900 nm). Embodiments of the invention include, for example, compounds having any one of formula (FX1 ) - (FX27), wherein A is a group corresponding to an indocyanine having one or more electron donating groups, electron withdrawing groups and/or photosensitizer groups as substituents.
[037] In an aspect, the invention provides compounds being of the formula (FX63) or (FX64):
Figure imgf000026_0001
Figure imgf000027_0001
(FX64); or a pharmaceutically acceptable salt or ester thereof; each of X1 to X17 is independently hydrogen, -OCF3, Ci-C6 alkyl, C3-C6 cycloalkyl, C5-C10 aryl, C5-C10 heteroaryl, CrC6 acyl, C2-C6 alkenyl, C2-C6 alkynyl, C5-C10 alkylaryl, halo, halomethyl, dihalomethyl, trihalomethyl, -C02R4°, -SOR41 , -OSR42, - SO2OR43, -CH2(CH20CH2)cCH2OH, -POaR^R45, -OR46, -SR47, -NR48R49, -NR50COR51 , -CN, -CONR52R53, -COR54, -NO2, -SO2R55, -S02NR56R57, PS1 or PS2; each of R40 to R57 is independently hydrogen, C C6 alkyl, C3-C6 cycloalkyl, C5-C10 aryl or C5-C10 heteroaryl; each of h and g is independently an integer selected from the range of from 1 to 3; and each of j and i is independently 0 or an integer selected from the range of from 1 to 10; and wherein ring Z, L1 to L3, W1 to W3, U, q, r, and x are as set forth in the description of formula (FX1 ) and (FX2).
[038] In an aspect, the invention provides compounds being of the formula (FX65), (FX66), (FX67) or (FX68):
Figure imgf000028_0001

Figure imgf000029_0001
(FX68); or a pharmaceutically acceptable salt or ester thereof; and wherein, L1 to L3, W1 to W3, X1 to X17, U, j, i, q, r, and x are as set forth in the description of formula (FX1 ), (FX2), (FX63) and (FX64). [039] In an aspect, the invention provides compounds being of the formula (FX69), (FX70), (FX71 ) or (FX72):
Figure imgf000030_0001
Figure imgf000031_0001
(FX72); or a pharmaceutically acceptable salt or ester thereof; and wherein, L1 to L3, W1 to W3, X1 to X17, U, j, i, q, r, and x are as set forth in the description of formula (FX1 ), (FX2), (FX63) and (FX64).
[040] In an aspect, the invention provides compounds being of the formula (FX73) or (FX74):
Figure imgf000032_0001
pharmaceutically acceptable salt or ester thereof; and wherein, L1 to L3, W1 to W3, X1 to X17, U, j, i, q, r, and x are as set forth in the description of formula (FX1 ), (FX2), (FX63) and (FX64).
[041] In an embodiment, the invention provides compounds having any one of formula (FX1 ) - (FX27), wherein A is a group corresponding to an azulene or azaazulene, for example an azulene or azaazulene providing a chromophore and/or fluorophore that absorbs
electromagnetic radiation having wavelengths in the visible and/or infrared regions of the electromagnetic spectrum (e.g., 350 nm to 1300 nm, or optionally 400 nm to 900 nm).
Embodiments of the invention include, for example, compounds having any one of formula
(FX1 ) - (FX27), wherein A is a group corresponding to an azulene or azaazulene having one or more electron donating groups, electron withdrawing groups and/or photosensitizer groups as substituents. [042] In an aspect, the invention provides compounds being of the formula (FX75), (FX76),
Figure imgf000033_0001
Figure imgf000034_0001
(FX78); or a pharmaceutically acceptable salt or ester thereof; wherein: G1 is -N-, -C(K)-, or -C(X1)-; G2 is -N-, -C(K)-, or - C(X2)-; G3 is -N-, -C(K)-, or -C(X3)-; G4 is -N-, -C(K)-, or -C(X4)-; G5 is -N-, -C(K)-, or - C(X5)-; G6 is -N-, -C(K)-, or -C(X6)-; G7 is -N-, -C(K)-, or -C(X7)-; G8 is -N-, -C(K)-, or - C(X8)-; wherein at most one of G1 - G8 is -N-; and wherein only one of G1 to G8 is -C(K)-; each K is a single bond to W1 , W2, or W3; each of X1 to X8 is independently hydrogen, -OCF3, Ci-C6 alkyl, C3-C6 cycloalkyl, C5-C10 aryl, C5-C10 heteroaryl, CrC6 acyl, C2-C6 alkenyl, C2-C6 alkynyl, C5-C10 alkylaryl, halo, halomethyl, dihalomethyl, trihalomethyl, -C02R4°, -SOR41 , - OSR42, -SO2OR43, -CH2(CH2OCH2)cCH2OH, -P03R44R45, -OR46, -SR47, -NR48R49, - NR50COR51 , -CN, -CONR52R53, -COR54, -N02, -S02R55, -S02NR56R57, PS1 or PS2; and each of R40 to R57 is independently hydrogen, CrC6 alkyl, C3-C6 cycloalkyl, C5-C10 aryl or C5-C10 heteroaryl. As used in this description -C(K)- indicates a direct or indirect linkage between an intra-ring carbon in the azulene or azaazulene group and the backbone of the compound. In some embodiments, the -C(K)- linkage is direct wherein the azulene or azaazulene group is directly bonded to ring Z (e.g., a phenyl or piperidine group) or a piperidine of the backbone. In some embodiments, the -C(K)- linkage is indirect wherein the azulene group or azaazulene group is bonded to W1-W3. In some embodiments, the -C(K)- linkage is indirect wherein the azulene group or azaazulene group is bonded to L1-L3. Reference to embodiments wherein K is a single bond to W1, W2, or W3 which is also designated as a single bond refers to direct linking of the azulene or azaazulene group to L1 , L2, or L3, if present, or direct linking of the azulene or azaazulene group to the piperidine group or ring Z of the backbone.
[043] In an aspect, the invention provides compounds being of the formula being of the formula (FX79) or (FX80):
(FX79) or
Figure imgf000035_0001
(FX80), or a pharmaceutically acceptable salt or ester thereof; wherein L1 to L3, W1 to W3, U, ring Z, G1 to G8, R1 - R5, x, r and q are as described in formulas (FX1 ), (FX2), (FX75), (FX76), (FX77) and (FX78).
[044] In an aspect, the invention provides compounds being of the formula being of the formula (FX81 ) or (FX82):
Figure imgf000035_0002
(FX81 )
or
Figure imgf000036_0001
(FX82); or a pharmaceutically acceptable salt or ester thereof; wherein L1 to L3, W1 to W3, U, ring Z, G1 to G8, R1 - R5, x, r and q are as described in formulas (FX1 ), (FX2), (FX75), (FX76), (FX77) and (FX78).
[045] In an aspect, the invention provides compounds being of the formula (FX83) or (FX84):
Figure imgf000036_0002
utically acceptable salt or ester thereof; wherein L1 to L3, W1 to W3, U, ring Z, G1 to G8, R1 - R5, x, r and q are as described in formulas (FX1 ), (FX2), (FX75), (FX76), (FX77) and (FX78).
[046] In an aspect, the invention provides compounds being of the formula (FX85), (FX86), (FX87) or (FX88):
Figure imgf000037_0001
pharmaceutically acceptable salt or ester thereof; wherein L1 to L3, W1 to W3, U, ring Z, G1 to G; R1 - R5, x, r and q are as described in formulas (FX1 ), (FX2), (FX75), (FX76), (FX77) and (FX78). [048] In an aspect, the invention provides compounds being of the formula (FX89), (FX90), (FX91 )
),
Figure imgf000038_0001
Figure imgf000039_0001
(FX92); or a pharmaceutically acceptable salt or ester thereof; wherein L1 to L3, W1 to W3, U, ring Z, G1 to G8, R1 - R5, x, r and q are as described in formulas (FX1 ), (FX2), (FX75), (FX76), (FX77) and (FX78).
[049] In an aspect, the invention provides compounds being of the formula (FX93), (FX94), (FX95)
Figure imgf000039_0002
Figure imgf000040_0001
acceptable salt or ester thereof; wherein L1 to L3, W1 to W3, U, ring Z, G1 to G8, R1 - R5, x, r and q are as described in formulas (FX1 ), (FX2), (FX75), (FX76), (FX77) and (FX78).
[050] In an aspect, the invention provides compounds being of the formula (FX97), (FX98),
Figure imgf000040_0002
Figure imgf000041_0001
(FX100); or a pharmaceutically acceptable salt or ester thereof; wherein L1 to L3, W1 to W3, U, ring Z, G1 to G8, R1 - R5, x, r and q are as described in formulas (FX1 ), (FX2), (FX75), (FX76), (FX77) and (FX78).
[051] In an aspect, the invention provides compounds being of the formula (FX101 ), (FX102), (FX103) or (FX104):
Figure imgf000042_0001
40
[052]
Figure imgf000043_0001
(FX104); or a pharmaceutically acceptable salt or ester thereof; wherein L1 to L3, W1 to W3, U, ring Z, G1 to G8, R1 - R5, x, r and q are as described in formulas (FX1 ), (FX2), (FX75), (FX76), (FX77) and (FX78).
[053] The present invention includes therapeutic agents useful for biomedical applications comprising purified stereoisomers (e.g., enantiomers and diastereomers), salts (including quarternary salts), and/or ionic forms (e.g., protonated and deprotonated forms) of the compounds of any of formula (FX1 ) - (FX113) , and mixtures thereof. As will be understood by those having general skill in the art, acidic functional groups and basic functional groups of the compounds of any of formula (FX1 ) - (FX113) may be in protonated or deprotonated states depending on the molecular environment (e.g., pH, ionic strength, composition, etc.), for example during synthesis, formulation and/or administration. For example, the invention includes, but is not limited to, the hydroxamate forms, and related salts, of all of the hydroxamic acids shown in formula (FX1 ) - (FX113).
[054] Selection of substituents of optical dye groups in the present compounds is useful for providing compounds exhibiting optical and physical properties beneficial for biomedical applications, including optical diagnostics, imaging, visualization and phototherapy. In certain embodiments of the invention, the composition of ring substituents on the pyrazine group (e.g., X1 to X3) or indocyanine group (e.g., X1 to X17) or azulene group (e.g., X1 to X8) or azaazulene group (e.g., X1 to X8) in compositions having formula (FX1) - (FX113) is selected to achieve preselected properties, such as optical, physiochemical and pharmacokinetic properties useful for biomedical applications. The invention provides, for example, compositions having any one of (FX1 ) - (FX113) wherein at least one of X1 to X3 or X1 to X17 X1 to X8 is an electron
withdrawing group (EWG) bonded directly or indirectly to a carbon atom of the pyrazine group or indocyanine group or azulene group or azaazulene group and at least one of X1 to X3 or X1 to X17 X1 to X8 is an electron donating group (EDG) bonded directly or indirectly to a carbon atom of the pyrazine group or indocyanine group or azulene group or azaazulene group.
Incorporation of a combination of an EWG and an EDG as substituents of different carbon atoms of the pyrazine group or indocyanine group or azulene group or azaazulene group is particularly beneficial for providing optical agents having large extinction coefficients in the visible and near infrared regions of the electromagnetic spectrum (e.g., 350 nm - 1300 nm, optionally 400 nm to 900 nm), emission in the visible and near infrared regions(e.g., 350 nm - 1300 nm, optionally 500 - 900 nm), a large fluorescence quantum yield (e.g., >0.1 ) and a Stoke's shift useful for optical detection and imaging (e.g., Stoke's shift > 10 nm). In some embodiments, for example, an electron withdrawing group and electron donating group are positioned on adjacent carbon atoms of the pyrazine group or indocyanine group or azulene group or azaazulene group. Alternatively, the invention includes embodiments wherein an electron withdrawing group and an electron donating group are positioned on non-adjacent carbon atoms of the pyrazine group or indocyanine group or azulene group or azaazulene group. Multiple electron withdrawing groups and/or electron donating groups on each substituent arm of the pyrazine group or indocyanine group or azulene group or azaazulene group are contemplated by the compositions of this aspect of the invention. By way of example, one EWG arm may comprise two, three, or more electron withdrawing groups bonded to the pyrazine or indocyanine or azulene group or azaazulene group core via a common linking moiety.
[055] In an embodiment, for example, the invention provides a compound having any one of formula (FX1 ) - (FX113), wherein at least one of X1 to X3 or X1 to X17 or X1 to X8 is an electron donating group, and wherein at least one of X1 to X3 or X1 to X17 or X1 to X8 is an electron withdrawing group. In an embodiment, for example, the invention provides a compound having any one of formula (FX1 ) - (FX113), wherein at least one of X1 to X3 or X1 to X17 or X1 to X8 is Ci-C6 alkyl, -OR46, -SR47, -NR48R49, or -NR50COR51 ; and wherein at least one of X1 to X3 or X1 to X17 or X1 to X8 is -CN, -C02R4°, -S02OR43, -CONR52R53, -COR54, -N02, -SOR41, -S02R55, -P03R44R45, halo, C C6 acyl, trihalomethyl, or -S02NR56R57. In an embodiment, for example, the invention provides a compound having any one of formula (FX1 ) - (FX113), wherein at least one of X1 to X3 or X1 to X17 or X1 to X8 is -OR46, -SR47, -NR48R49, or -NR50COR51. In an embodiment, for example, the invention provides a compound having any one of formula (FX1 ) - (FX113), wherein at least one of X1 to X3 or X1 to X17 or X1 to X8 is -NR48R49, or -NR50COR51. In an embodiment, for example, the invention provides a compound having any one of formula (FX1 ) - (FX113), wherein at least one of X1 to X3 or X1 to X17 or X1 to X8 is -CN, -C02R40,- COR54, -N02, -S02R55, or -S02NR56R57. In an embodiment, for example, the invention provides a compound having any one of formula (FX1 ) - (FX113), wherein at least one of X1 to X3 or X1 to X17 or X1 to X8 is -C02R40, -COR54, -S02NR56R57 or -S02R55. In an embodiment, for example, the invention provides a compound having any one of formula (FX1 ) - (FX113), wherein at least one of X1 to X3 or X1 to X17 or X1 to X8 is -NR48R49, and wherein at least one of X1 to X3 or X1 to X17 or X1 to X8 is -CO2R40,-COR54, -S02NR56R57 or -S02R55. [056] In an embodiment, the present invention provides compositions having any one of formula (FX28) - (FX62), wherein at least one of X1, X2 and X3 is -OR46, -SR47, -NR48R49, and - NR50COR51. In an embodiment, the present invention provides compositions having any one of formula (FX28) - (FX62), wherein at least one of X1, X2 and X3 is -NR48R49, or -NR50COR51. In an embodiment, the present invention provides compositions having any one of formula (FX28) - (FX62), wherein at least one of X1, X2 and X3 is -CN, -CO^ -COR54, -N02, -S02R55, or— S02NR56R57. In an embodiment, the present invention provides compositions having any one of formula (FX28) - (FX62), wherein at least one of X1, X2 and X3 is -COzR^ -COR54,—
S02NR56R57 or -S02R55. In an embodiment, the present invention provides compositions having any one of formula (FX28) - (FX62), wherein at least one of X1, X2 and X3 is -NR48R49 and wherein at least one of X1, X2 or X3 is -CO2R40,-COR54, -S02NR56R57 or -S02R55.
[057] In an embodiment, the invention provides compounds with electron-donating and electron-withdrawing groups attached to adjacent positions of a pyrazine optical dye component. In an embodiment, provided are compounds of formula (FX28) - (FX62) wherein:
(a) any one of X1 and X2 is C C6 alkyl, -OR46, -SR47, -NR48R49, or -NR50COR51 and the other of X1 and X2 is -CN, -C02R40, -S02OR43, -CONR52R53, -COR54, -N02, -SOR41 , -S02R55, - POsR^R45, halo, C C6 acyl, trihalomethyl, or -S02NR56R57;
(b) any one of X1 and X3 is C C6 alkyl, -OR46, -SR47, -NR48R49, or -NR50COR51 and the other of X1 and X3 is -CN, -C02R40, -S02OR43, -CONR52R53, -COR54, -N02, -SOR41 , -S02R55, - POsR^R45, halo, C C6 acyl, trihalomethyl, or -S02NR56R57;
(c) any one of X2 and X3 is C C6 alkyl, -OR46, -SR47, -NR48R49, or -NR50COR51 and the other of X2 and X3 is -CN, -C02R40, -S02OR43, -CONR52R53, -COR54, -N02, -SOR41 , -S02R55, - POsR^R45, halo, C C6 acyl, trihalomethyl, or -S02NR56R57;
(d) any two of X1, X2 and X3 is C C6 alkyl, -OR46, -SR47, -NR48R49, or -NR50COR51 and the other of X1, X2 and X3 is -CN, -C02R40, -S02OR43, -CONR52R53, -COR54, -N02, -SOR41, - S02R55, -POsR^R45, halo, C C6 acyl, trihalomethyl, or -S02NR56R57; or
(e) any two of X1, X2 and X3 is -CN, -C02R40, -S02OR43, -CONR52R53, -COR54, -N02, -SOR41 , -S02R55, -P03R44R45, halo, C C6 acyl, trihalomethyl, or -S02NR56R57 and the other of X1, X2 and X3 is C C6 alkyl, -OR46, -SR47, -NR48R49, or -NR50COR51 .
[058] In an embodiment, a compound of the present invention has any one of formula (FX1 ) - (FX113), wherein each of R1 and R5 is hydrogen. In an embodiment, a compound of the present invention has any one of formula (FX1 ) - (FX113), wherein each of R1, R2, R4 and R5 is hydrogen. In an embodiment, a compound of the present invention has any one of formula
(FX1 ) - (FX113), wherein each of R1 to R5 is hydrogen. In an embodiment, a compound of the present invention has any one of formula (FX1 ) - (FX113), wherein R3 is a group other than hydrogen. In an embodiment, a compound of the present invention has any one of formula
(FX1 ) - (FX113), wherein R3 is -OCF3. In an embodiment, a compound of the present invention has any one of formula (FX1) - (FX113), wherein R3 is PS1 or PS2, and optionally each of R1 - R2 and R4 - R5 is hydrogen. In an embodiment, a compound of the present invention has any one of formula (FX1 ) - (FX113) wherein at least one of X1 to X3 or X1 to X17 or X1 to X8 is PS1 or PS2.
[059] A variety of linking moieties (e.g., W1, W2 and/ or W3) may be used to covalently link the sulfone hydroxamic acid or hydroxamate backbone and the optical dye component of compounds of the present invention. Some embodiments incorporate an optional spacer moiety (e.g., L1, L2 and/ or L3) between the sulfone hydroxamic acid or hydroxamate backbone and the optical dye component, for example, to ensure that the molecular recognition functionality of the sulfone hydroxamic acid or hydroxamate backbone is retained and/or to preselect or optimize the optical functionality of the optical dye component. In an embodiment, the invention provides compounds having any one of formula (FX1 ) - (FX113), wherein at least one of W1 and L1, W2 and L2, or form: -(CH2)n- -(CH2)mNHCO- -CONH(CH2)nCO- , -
0(CH2)n-
Figure imgf000046_0001
wherein n and m are as set forth in the description of formulas
(FX1 ) and (FX2). In an embodiment, the invention provides compounds having any one of formula (FX1 ) - (FX113), wherein at least one of W1 and L1, W2 and L2, or W3 and L3 combine to form
Figure imgf000046_0002
Figure imgf000046_0003
wherein n and a are as set forth in the description of formulas (FX1 ) and (FX2).
[060] In an embodiment, the invention provides a compound having any of formula (FX1 ) - (FX113), wherein each of R1 to R5 and R8 to R10 is hydrogen, Ci - C6 alkyl or -OCF3, and optionally for some embodiments wherein each of R1 to R5 and R8 to R10 is hydrogen or Ci - C6 alkyl, and optionally for some embodiments wherein each of R1 to R5 and R8 to R10 is hydrogen or Ci - C3 alkyl, and optionally for some embodiments wherein each of R1 to R5 and R8 to R10 is hydrogen. In an embodiment, the invention provides a compound having any of formula (FX1 ) - (FX113), wherein at least one of R1 to R5 and R8 to R10 is -OCF3, and optionally wherein R3 is - OCF3. In an embodiment, the invention provides a compound having any of formula (FX1 ) - (FX113), wherein at least one of R1 to R5 and R8 to R10 is PS1. In an embodiment, the invention provides a compound having any of formula (FX1 ) - (FX113), wherein at least one of R1 to R5 and R8 to R10 is PS2. In an embodiment, the invention provides a compound having any of formula (FX1 ) - (FX113), wherein R1 is hydrogen; and/or R2 is hydrogen; and/or R4 is hydrogen; and/or R5 is hydrogen. In an embodiment, the invention provides a compound having any of formula (FX1 ) - (FX113), wherein U is -O- or -S- and optionally for some embodiments wherein U is -0-. In an embodiment, the invention provides a compound having any of formula (FX1 ) - (FX113), wherein each of R11 to R34 and R60 to R79 is independently hydrogen or CrC6 alkyl; and optionally wherein each of R11 to R34 and R60 to R79 is
independently hydrogen or C-|-C3 alkyl; and optionally wherein each of R11 to R34 and R60 to R79 is independently hydrogen. In an embodiment, the invention provides a compound having any of formula (FX63) - (FX74), wherein each of X1 to X17 is hydrogen, an electron donating group or an electron withdrawing group, and optionally wherein at least two of X1 to X17 is an electron donating group or an electron withdrawing group and the rest of X1 to X17 is hydrogen. In an embodiment, the invention provides a compound having any of formula (FX28) - (FX62), wherein each of X1 to X3 is hydrogen, an electron donating group or an electron withdrawing group, and optionally wherein at least two of X1 to X3 is an electron donating group or an electron withdrawing group and the rest of X1 to X3 is hydrogen. In an embodiment, the invention provides a compound having any of formula (FX75) - (FX104), wherein each of X1 to X8 is hydrogen, an electron donating group or an electron withdrawing group, and optionally wherein at least two of X1 to X8 is an electron donating group or an electron withdrawing group and the rest of X1 to X8 is hydrogen. In an embodiment, the invention provides a compound having any of formula (FX1 ) - (FX113), wherein each of each of R40 to R57 is hydrogen or C-|-C6 alkyl, and optionally wherein each of each of R40 to R57 is hydrogen. In an embodiment, the invention provides a compound having any of formula (FX1 ) - (FX113), wherein Y is -CR2-, and optionally wherein Y is -CH-. In an embodiment, the invention provides a compound having any of formula (FX1 ) - (FX113), wherein Y is -N-. In an embodiment, the invention provides a compound having any of formula (FX1 ) - (FX113), wherein each of h and g are 1 or 2, and optionally wherein each of h and g are 1.
[061] In an embodiment, a compound of the invention having any of formula (FX1 ) - (FX113) selectively binds to one or more MMPs, preferably for some applications under in vivo conditions. In certain embodiments compounds of the present invention are selective with respect to which MMP they bind to or otherwise associate with. In some embodiments, for example, the invention provides compounds that bind to a single MMP or a group of selected MMPs, such as MMP-2, MMP-9 and/or MMP-13. In an embodiment, for example, the invention provides compounds that bind to some MMPs and do not bind with one or more selected MMPs, such as MMP-1 . In an embodiment, the compounds provided are MMP selective compounds. Without being bound by theory, it is believed the sulfone hydroxamic acid or hydroxamate backbone described herein selectively binds to MMPs, for example MMP-2 and MMP-9. In an embodiment, a compound provided has a sulfone hydroxamic acid or hydroxamate backbone that binds selectively to one or more MMPs. In an embodiment, the invention provides compounds having any of formula (FX1 ) - (FX113) that inhibit the biological activity of one or more MMPs, for example by inhibiting biological activity associated with an MMP or a class of MMPs involved in degradation and/or remodeling of the extracellular matrix. In an embodiment, a compound provided selectively binds to a related metalloproteinase called TNF-a Converting Enzyme (TACE) which is implicated in the cartilage catabolism pathway.
[062] In some embodiments, compounds of the invention may optionally include a poly(ethylene glycol) (abbreviated as PEG) component. In an embodiment, for example, the invention provides a composition having any one of the formula (FX1) - (FX113), wherein at least one of W1 to W3, L1 to L3, R1 to R5 and R8 to R10 is a substituent comprising -(CH2OCH2) m-, or a derivative thereof, wherein m is selected from the range of 1 to 100. Incorporation of a poly(ethylene glycol) component in some compositions of the invention provides
pharmacokinetic, chemical, and/or physical properties useful for bioanalytical, diagnostic and/or therapeutic applications. Poly(ethylene glycol) containing compounds of some embodiments of the present invention, for example, provide enhanced biocompatibility, low toxicity and suppress immune responses upon administration. Poly(ethylene glycol) containing compounds of some embodiments of the invention facilitate formulation, administration and/or delivery, for example, by enhancing solubility.
[063] In an embodiment, the invention provides a compound being of the formula (FX105),
Figure imgf000048_0001
Figure imgf000049_0001
aceutically acceptable salt or ester thereof.
[066] The invention further provides a compound having any one of formula (FX1 ) - (FX113), or a pharmaceutical formulation thereof, for use in an optical imaging, diagnostic, and/or therapeutic biomedical procedure. In an embodiment, the invention provides an optical agent comprising a pharmaceutically acceptable formulation, wherein at least one active ingredient of the formulation is a compound having any one of formula (FX1 ) - (FX113) provided in a therapeutically or diagnostically effective amount. The invention includes, for example, formulations comprising a compound having any one of formula (FX1 ) - (FX113) and one or more pharmaceutically acceptable carriers or excipients. In an embodiment, the biomedical procedure comprises: (i) administering to a subject a therapeutically or diagnostically effective amount of the compound having any one of formula (FX1 ) - (FX113) under conditions sufficient for contacting a target tissue and/or target cell with the compound, wherein the compound selectively binds to matrix metalloproteinase enzyme expressed by, or otherwise associated with, the target tissue or target cell (e.g., target tissue characterized by an elevated
concentration of MMPs greater than in target tissue in a normal/healthy state); and optionally (ii) exposing the administered compound to a therapeutically or diagnostically effective amount of electromagnetic radiation. In an embodiment, the compound is administered to the subject in an amount sufficient to inhibit the biological activity of MMPs expressed or otherwise generated by the target tissue or target cells. In an embodiment, the compound is administered to the subject in an amount sufficient to suppress or otherwise attenuate biological processes associated with MMPs in the target tissue, such as the degradation and/or remodeling of the extra cellular matrix. In an embodiment, the compound is administered to the tissue of a tumor of the subject in an amount sufficient to suppress or prevent the growth, proliferation and/or metastasis of the tumor.
[067] In an embodiment, the administered compound is exposed at the site of the target tissue or target cell to electromagnetic radiation having wavelengths selected over a range of 350 nanometers to 1300 nanometers, optionally having wavelengths selected over a range of 350 nanometers to 900 nanometers. In an embodiment, exposing the administered compound to electromagnetic radiation generates fluorescence, wherein the biomedical procedure further comprises detecting fluorescence from the administered compound. In an embodiment, exposing the administered compound to electromagnetic radiation generates a diagnostically effective amount of fluorescence, for example an amount of fluorescence allowing for optical detection and/or imaging of the target tissue . In an embodiment, a method of the invention further comprises exposing the administered compound at the target tissue to electromagnetic radiation having sufficient power, fluence, intensity and/or dose (net number of photons provided to the target tissue) to provide optical detection and/or imagining of the target tissue. In an embodiment, a method of the invention further comprises exposing the administered compound at the target tissue to a diagnostically effective and/or therapeutically effective amount of electromagnetic radiation. In an embodiment, a method of the invention further comprises generating an image of the fluorescence from the compound. In an embodiment, a method of the invention further comprises visualizing the fluorescence from the compound. In an embodiment, a method of the invention further comprises measuring the intensity, energy or power of fluorescence from the compound, optionally as a function of illuminated area.
[068] In an embodiment, the biomedical procedure comprises administering or otherwise targeting the administered compound to a target tissue or target cell of the subject, such as a tumor, lesion, site of inflammation, vasculature tissue, or an organ, wherein the target tissue or target cell expresses or otherwise generates MMPs. In an embodiment, for example, the target tissue is a tissue type selected from the group consisting of colon, prostate, gastric, esophageal, uterine, endometrial, pancreatic, breast, cervical, brain, skin, gallbladder, lung, heart, central nervous system and ovary.
[069] Another aspect of the invention is directed to methods of using the compounds of any one of formulas (FX1 ) - (FX113) in biomedical procedures including treatment and diagnosis of diseases and other pathological conditions. The invention includes, for example, methods of using compounds of any one of formulas (FX1 ) - (FX113) in a biomedical procedure, including photodiagnostic and therapeutic methods such as optical imaging, anatomical visualization, endoscopic visualization, image guided surgery, and optical biopsy of tumors and other lesions. The invention includes, for example, methods of using compounds of any one of formulas (FX1 ) - (FX113) in a therapeutic procedure involving inhibition of the biological activity of MMPs in the target tissue. In one such biomedical procedure, a therapeutically effective amount of a compound of any one of formulas (FX1 ) - (FX113) is administered to a subject (e.g., via intravenous or intraarterial injection, oral administration, topical administration, etc.). A method of the present invention further comprises exposing the administered compound to a
diagnostically effective amount of electromagnetic radiation, such as electromagnetic radiation having wavelengths in the visible and near infrared regions of the electromagnetic spectrum (e.g., 350 nm to 1300 nm). In a method, the electromagnetic radiation exposed to the compound of any one of formulas (FX1 ) - (FX113) does not have wavelengths in the X-ray region of the electromagnetic spectrum. In a method, the electromagnetic radiation exposed to the compound of any one of formulas (FX1 ) - (FX113) does not have wavelengths in the ultraviolet region of the electromagnetic spectrum. In an embodiment, non-ionizing
electromagnetic radiation is used in the present methods. "Non-ionizing electromagnetic radiation" herein refers to electromagnetic radiation wherein a single photon does not have enough energy to completely remove at least one electron from an atom or molecule of the subject's body.
[070] In an embodiment, the present invention provides methods for treating cancer or a cancer-related disorder. A method includes administering a therapeutically effective amount of a compound having any one of formulas (FX1 ) - (FX113) to a subject in need of treatment for cancer. In various embodiments, these cancers may include colorectal, prostate, gastric, esophageal, uterine-endometrial, pancreatic, breast, cervical, head and neck, hepatic, skin, gallbladder, lung, and ovarian cancers. In an embodiment, the present invention provides methods of treating inflammation or inflammation-associated disorders, the method including administering a therapeutically-effective amount of a compound having any one of formulas (FX1 ) - (FX113) to a subject in need of treatment for inflammation or an inflammation- associated disorder. In various embodiments, these disorders may include, but are not limited to rheumatoid arthritis; osteoarthritis; and septic arthritis.
[071] The invention further provides a compound having any one of formula (FX1 ) - (FX113), or a pharmaceutical formulation thereof, for use in a medical phototherapy procedure, such as a Type 1 or Type 2 phototherapy procedure. In an embodiment of this aspect, a compound of the invention has any one of formula (FX1) - (FX113), wherein at least one of R1 to R5 and X1 to X3 or X1 to X17 or X1 to X8 is PS1 or PS2. In an embodiment, the invention provides a
phototherapeutic agent comprising a pharmaceutically acceptable formulation, wherein an active ingredient of the formulation provided in a therapeutically effective amount is a compound having any one of formula (FX1 ) - (FX113). The invention includes, for example, formulations comprising a compound having any one of formula (FX1 ) - (FX113) and one or more
pharmaceutically acceptable carriers or excipients. In an embodiment, the medical
phototherapy procedure comprises: (i) administering to a subject in need of treatment a therapeutically effective amount of the compound having any one of formula (FX1 ) - (FX113); and (ii) exposing the administered compound to electromagnetic radiation. In an embodiment, the administered compound is exposed to electromagnetic radiation having wavelengths selected over a range of 350 nanometers to 1300 nanometers, optionally having wavelengths selected over a range of 350 nanometers to 900 nanometers. In an embodiment, exposing the administered compound to electromagnetic radiation generates one or more radicals, nitrenes, carbenes, ions, and/or singlet oxygen. In an embodiment, exposing the administered compound to electromagnetic radiation generates a therapeutically effective amount of photoactivated compound. In an embodiment, exposing the administered compound to electromagnetic radiation generates a therapeutically effective amount of reactive species causing localized cell death or injury. In an embodiment, the medical phototherapy procedure comprises
administering or otherwise targeting the administered compound to a target tissue of the subject, such as a tumor , lesion, site of inflammation, vasculature tissue, or organ. In an embodiment, methods of the invention further comprises exposing the administered compound at the target tissue to light having sufficient power, fluence, intensity and/or dose (net number of photons provided to the target tissue) to result in injury, inactivation and/or death to cells at the target tissue.
[072] The invention includes, for example, methods of using the compounds of any one of formulas (FX1 ) - (FX113) in a medical phototherapy procedure. In one such medical phototherapy procedure, a therapeutically effective amount of a compound of any one of formulas (FX1 ) - (FX113) is administered to a subject (e.g., via intravenous or intraarterial injection, oral administration, topical administration, etc.) and exposed to a therapeutically effective amount of electromagnetic radiation, such as electromagnetic radiation having wavelengths in the visible and near infrared regions of the electromagnetic spectrum (e.g., 3500 nm to 1300 nm). In a method, the electromagnetic radiation exposed to the compound of any one of formulas (FX1 ) - (FX113) does not have wavelengths in the X-ray region of the electromagnetic spectrum. In a method, the electromagnetic radiation exposed to the compound of any one of formulas (FX1 ) - (FX113) does not have wavelengths in the ultraviolet region of the electromagnetic spectrum.
BRIEF DESCRIPTION OF THE FIGURES
[073] Figure 1 provides the response curve for inhibition of MMP-2 by compound (FX107). In this Figure, percentage inhibition is plotted as a function of concentration (μΜ).
[074] Figure 2 provides the response curve for inhibition of MMP-9 by compound (FX107). In this Figure, percentage inhibition is plotted as a function of concentration (μΜ). [075] Figure 3 provides the response curve for inhibition of MMP-2 by compound (FX108).
In this Figure, percentage inhibition is plotted as a function of concentration (μΜ).
[076] Figure 4 provides the response curve for inhibition of MMP-2 by compound (FX105).
In this Figure, percentage inhibition is plotted as a function of concentration (μΜ).
[077] Figure 5 provides the response curve for inhibition of MMP-9 by compound (FX105).
In this Figure, percentage inhibition is plotted as a function of concentration (μΜ).
[078] Figure 6A provides a high resolution mass spectrum of the compound having formula
(FX105). Figure 6B provides an expanded view of the mass spectrum (top panel) of the compound having formula (FX105) and a simulated mass spectrum (lower panel) for a mass to charge ratio (m/z) range of about 1 149 to about 1 154. Figure 6C provides an expanded view of the mass spectrum (top panel) of the compound having formula (FX105) and a simulated mass spectrum (lower panel) for a mass to charge ratio (m/z) range of about 574 to 576.
STATEMENTS REGARDING CHEMICAL COMPOUNDS AND NOMENCLATURE
[079] In an embodiment, a composition or compound of the invention is isolated or purified. In an embodiment, an isolated or purified compound is at least partially isolated or purified as would be understood in the art. In an embodiment, the composition or compound of the invention has a chemical purity of 95%, optionally for some applications 99%, optionally for some applications 99.9%, optionally for some applications 99.99%, and optionally for some applications 99.999% pure.
[080] Many of the molecules disclosed herein contain one or more ionizable groups, lonizable groups include groups from which a proton can be removed (e.g., -COOH, S03H, etc.) or added (e.g., amines) and groups which can be quaternized (e.g., amines). All possible ionic forms of such molecules and salts thereof are intended to be included individually in the disclosure herein. With regard to salts of the compounds herein, one of ordinary skill in the art can select from among a wide variety of available counterions that are appropriate for preparation of salts of this invention for a given application. In specific applications, the selection of a given anion or cation for preparation of a salt can result in increased or decreased solubility of that salt. For example, indication of the functional group -COOH is intended to also include the deprotonated form of this group, e.g., -COO". For example, indication of the functional group -SO3H is intended to also include the deprotonated form of this group, e.g., -S03 ".
[081 ] The compounds of this invention can contain one or more chiral centers. Accordingly, this invention is intended to include racemic mixtures, diasteromers, enantiomers, tautomers and mixtures enriched in one or more stereoisomer. The scope of the invention as described and claimed encompasses the racemic forms of the compounds as well as the individual enantiomers and non-racemic mixtures thereof.
[082] As used herein, the term "group" may refer to a functional group of a chemical compound. Groups of the present compounds refer to an atom or a collection of atoms that are a part of the compound. Groups of the present invention may be attached to other atoms of the compound via one or more covalent bonds. Groups may also be characterized with respect to their valence state. The present invention includes groups characterized as monovalent, divalent, trivalent, etc. valence states.
[083] As is customary and well known in the art, hydrogen atoms in formulas (FX1 ) - (FX113) are not always explicitly shown, for example, hydrogen atoms bonded to the carbon atoms of aromatic, heteroaromatic, and alicyclic rings are not always explicitly shown in formulas (FX1 ) - (FX113). The structures provided herein, for example in the context of the description of formulas (FX1 ) - (FX113), are intended to convey to one of reasonable skill in the art the chemical composition of compounds of the methods and compositions of the invention, and as will be understood by one of skill in the art, the structures provided do not indicate the specific positions of atoms and bond angles between atoms of these compounds.
[084] As used herein, the symbol convention "-X-," wherein X is an atom or functional group, refers generally to a bonding configuration wherein X is bonded to two other adjacent atoms and/or functional groups. In some embodiments, -X- refers to a linking group or spacer group, including linear and branched groups. In some embodiments, -X- refers to an intra-ring member, for example, an intra-ring member of a cycloalkyl group, aryl group, or heteroaryl group. The designation "-X-" does not specify the type of bond (single, double, triple, etc.) that X forms to the adjacent atoms and/or functional groups. In the context of formulas (FX75) - (FX78), for example, the symbol -N- in the context of G1-G8 refers to azaazulene containing- compounds having an intra-ring nitrogen atom bonded to adjacent intra-ring carbon atoms. In the context of formulas (FX75) - (FX78), for example, the symbol -C(Xn)-, wherein n is an integer from 1 to 8, in the context of G1-G8 refers to azulene or azaazulene containing- compounds having an intra-ring carbon atom bonded to adjacent intra-ring carbon and/or nitrogen atoms and also bound to substituent Xn.
[085] As used herein, the terms "alkylene" and "alkylene group" are used synonymously and refer to a divalent group derived from an alkyl group as defined herein. The invention includes compounds having one or more alkylene groups. Alkylene groups in some compounds function as attaching and/or spacer groups. Compounds of the invention may have substituted and/or unsubstituted C C2o alkylene, CrC10 alkylene and C C5 alkylene groups.
[086] As used herein, the terms "cycloalkylene" and "cycloalkylene group" are used synonymously and refer to a divalent group derived from a cycloalkyl group as defined herein. The invention includes compounds having one or more cycloalkylene groups. Cycloalkyl groups in some compounds function as attaching and/or spacer groups. Compounds of the invention may have substituted and/or unsubstituted C3-C2o cycloalkylene, C3-C10 cycloalkylene and C3-C5 cycloalkylene groups.
[087] As used herein, the terms "arylene" and "arylene group" are used synonymously and refer to a divalent group derived from an aryl group as defined herein. The invention includes compounds having one or more arylene groups. In some embodiments, an arylene is a divalent group derived from an aryl group by removal of hydrogen atoms from two intra-ring carbon atoms of an aromatic ring of the aryl group. Arylene groups in some compounds function as attaching and/or spacer groups. Arylene groups in some compounds function as chromophore, fluorophore, aromatic antenna, dye and/or imaging groups. Compounds of the invention include substituted and/or unsubstituted C3-C30 arylene, C3-C2o arylene, C3-C10 arylene and C C5 arylene groups.
[088] As used herein, the terms "heteroarylene" and "heteroarylene group" are used synonymously and refer to a divalent group derived from a heteroaryl group as defined herein. The invention includes compounds having one or more heteroarylene groups. In some embodiments, a heteroarylene is a divalent group derived from a heteroaryl group by removal of hydrogen atoms from two intra-ring carbon atoms or intra-ring nitrogen atoms of a heteroaromatic or aromatic ring of the heteroaryl group. Heteroarylene groups in some compounds function as attaching and/or spacer groups. Heteroarylene groups in some compounds function as chromophore, aromatic antenna, fluorophore, dye and/or imaging groups. Compounds of the invention include substituted and/or unsubstituted C3-C30 heteroarylene, C3-C2o heteroarylene, C1-C10 heteroarylene and C3-C5 heteroarylene groups.
[089] As used herein, the terms "alkenylene" and "alkenylene group" are used synonymously and refer to a divalent group derived from an alkenyl group as defined herein. The invention includes compounds having one or more alkenylene groups. Alkenylene groups in some compounds function as attaching and/or spacer groups. Compounds of the invention include substituted and/or unsubstituted C2-C2o alkenylene, C2-Ci0 alkenylene and C2-C5 alkenylene groups.
[090] As used herein, the terms "cylcoalkenylene" and "cylcoalkenylene group" are used synonymously and refer to a divalent group derived from a cylcoalkenyl group as defined herein. The invention includes compounds having one or more cylcoalkenylene groups. Cycloalkenylene groups in some compounds function as attaching and/or spacer groups. Compounds of the invention include substituted and/or unsubstituted C3-C20 cylcoalkenylene, C3-C10 cylcoalkenylene and C3-C5 cylcoalkenylene groups.
[091 ] As used herein, the terms "alkynylene" and "alkynylene group" are used synonymously and refer to a divalent group derived from an alkynyl group as defined herein. . The invention includes compounds having one or more alkynylene groups. Alkynylene groups in some compounds function as attaching and/or spacer groups. Compounds of the invention include substituted and/or unsubstituted C2-C20 alkynylene, C2-C10 alkynylene and C2-C5 alkynylene groups.
[092] As used herein, the term "halo" refers to a halogen group such as a fluoro (-F), chloro (-CI), bromo (-Br), iodo (-I) or astato (-At). [093] The term "heterocyclic" refers to ring structures containing at least one other kind of atom, in addition to carbon, in the ring. Examples of such atoms include nitrogen, oxygen and sulfur. Examples of heterocyclic rings include, but are not limited to, pyrrolidinyl, piperidyl, imidazolidinyl, tetrahydrofuryl, tetrahydrothienyl, furyl, thienyl, pyridyl, quinolyl, isoquinolyl, pyridazinyl, pyrazinyl, indolyl, imidazolyl, oxazolyl, thiazolyl, pyrazolyl, pyridinyl, benzoxadiazolyl, benzothiadiazolyl, triazolyl and tetrazolyl groups. Atoms of heterocyclic rings can be bonded to a wide range of other atoms and functional groups, for example, provided as substituents.
[094] The term "carbocyclic" refers to ring structures containing only carbon atoms in the ring. Carbon atoms of carbocyclic rings can be bonded to a wide range of other atoms and functional groups, for example, provided as substituents..
[095] The term "alicyclic" refers to a ring that is not an aromatic ring. Alicyclic rings include both carbocyclic and heterocyclic rings.
[096] As used herein, the term "alkoxyalkyl" refers to a substituent of the formula alkyl-O- alkyl.
[097] As used herein, the term "polyhydroxyalkyl" refers to a substituent having from 2 to 12 carbon atoms and from 2 to 5 hydroxyl groups, such as the 2,3-dihydroxypropyl, 2,3,4- trihydroxybutyl or 2,3,4,5-tetrahydroxypentyl residue.
[098] As used herein, the term "polyalkoxyalkyl" refers to a substituent of the formula alkyl- (alkoxy)n-alkoxy wherein n is an integer from 1 to 10, preferably 1 to 4, and more preferably for some embodiments 1 to 3.
[099] Amino acids include glycine, alanine, valine, leucine, isoleucine, methionine, proline, phenylalanine, tryptophan, asparagine, glutamine, glycine, serine, threonine, serine, Threonine, asparagine, glutamine, tyrosine, cysteine, lysine, arginine, histidine, aspartic acid and glutamic acid. As used herein, reference to "a side chain residue of a natural a-amino acid" specifically includes the side chains of the above-referenced amino acids.
[0100] AlkyI groups include straight-chain, branched and cyclic alkyl groups. AlkyI groups include those having from 1 to 30 carbon atoms. Alkyl groups include small alkyl groups having 1 to 3 carbon atoms. Alkyl groups include medium length alkyl groups having from 4-10 carbon atoms. Alkyl groups include long alkyl groups having more than 10 carbon atoms, particularly those having 10-30 carbon atoms. The term cycloalkyl specifically refers to an alky group having a ring structure such as ring structure comprising 3-30 carbon atoms, optionally 3-20 carbon atoms and optionally 2 - 10 carbon atoms, including an alkyl group having one or more rings. Cycloalkyl groups include those having a 3-, 4-, 5-, 6-, 7-, 8-, 9- or 10-member carbon ring(s) and particularly those having a 3-, 4-, 5-, 6-, or 7-member ring(s). The carbon rings in cycloalkyl groups can also carry alkyl groups. Cycloalkyl groups can include bicyclic and tricycloalkyl groups. Alkyl groups are optionally substituted. Substituted alkyl groups include among others those which are substituted with aryl groups, which in turn can be optionally substituted. Specific alkyl groups include methyl, ethyl, n-propyl, iso-propyl, cyclopropyl, n-butyl, s-butyl, t-butyl, cyclobutyl, n-pentyl, branched-pentyl, cyclopentyl, n-hexyl, branched hexyl, and cyclohexyl groups, all of which are optionally substituted. Substituted alkyl groups include fully halogenated or semihalogenated alkyl groups, such as alkyl groups having one or more hydrogens replaced with one or more fluorine atoms, chlorine atoms, bromine atoms and/or iodine atoms. Substituted alkyl groups include fully fluorinated or semifluorinated alkyl groups, such as alkyl groups having one or more hydrogens replaced with one or more fluorine atoms. An alkoxy group is an alkyl group that has been modified by linkage to oxygen and can be represented by the formula R-0 and can also be referred to as an alkyl ether group. Examples of alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy, butoxy and heptoxy. Alkoxy groups include substituted alkoxy groups wherein the alky portion of the groups is substituted as provided herein in connection with the description of alkyl groups. As used herein MeO- refers to CH3O-.
[0101] Alkenyl groups include straight-chain, branched and cyclic alkenyl groups. Alkenyl groups include those having 1 , 2 or more double bonds and those in which two or more of the double bonds are conjugated double bonds. Alkenyl groups include those having from 2 to 20 carbon atoms. Alkenyl groups include small alkenyl groups having 2 to 3 carbon atoms. Alkenyl groups include medium length alkenyl groups having from 4-10 carbon atoms. Alkenyl groups include long alkenyl groups having more than 10 carbon atoms, particularly those having 10-20 carbon atoms. Cycloalkenyl groups include those in which a double bond is in the ring or in an alkenyl group attached to a ring. The term cycloalkenyl specifically refers to an alkenyl group having a ring structure, including an alkenyl group having a 3-, 4-, 5-, 6-, 7-, 8-, 9- or 10-member carbon ring(s) and particularly those having a 3-, 4-, 5-, 6- or 7-member ring(s). The carbon rings in cycloalkenylgroups can also carry alkyl groups. Cycloalkenylgroups can include bicyclic and tricyclic alkenyl groups. Alkenyl groups are optionally substituted. Substituted alkenyl groups include among others those which are substituted with alkyl or aryl groups, which groups in turn can be optionally substituted. Specific alkenyl groups include ethenyl, prop-1-enyl, prop- 2-enyl, cycloprop-1 -enyl, but-1-enyl, but-2-enyl, cyclobut-1-enyl, cyclobut-2-enyl, pent-1-enyl, pent-2-enyl, branched pentenyl, cyclopent-1 -enyl, hex-1-enyl, branched hexenyl, cyclohexenyl, all of which are optionally substituted. Substituted alkenyl groups include fully halogenated or semihalogenated alkenyl groups, such as alkenyl groups having one or more hydrogens replaced with one or more fluorine atoms, chlorine atoms, bromine atoms and/or iodine atoms. Substituted alkenyl groups include fully fluorinated or semifluorinated alkenyl groups, such as alkenyl groups having one or more hydrogen atoms replaced with one or more fluorine atoms.
[0102] Aryl groups include groups having one or more 5-, 6- or 7- member aromatic and/or heterocyclic aromatic rings. The term heteroaryl specifically refers to aryl groups having at least one 5-, 6- or 7- member heterocyclic aromatic rings. Aryl groups can contain one or more fused aromatic and heteroaromatic rings or a combination of one or more aromatic or heteroaromatic rings and one or more nonaromatic rings that may be fused or linked via covalent bonds. Heterocyclic aromatic rings can include one or more N, O, or S atoms in the ring. Heterocyclic aromatic rings can include those with one, two or three N atoms, those with one or two O atoms, and those with one or two S atoms, or combinations of one or two or three N, O or S atoms. Aryl groups are optionally substituted. Substituted aryl groups include among others those which are substituted with alkyl or alkenyl groups, which groups in turn can be optionally substituted. Specific aryl groups include phenyl, biphenyl groups, pyrrolidinyl, imidazolidinyl, tetrahydrofuryl, tetrahydrothienyl, furyl, thienyl, pyridyl, quinolyl, isoquinolyl, pyridazinyl, pyrazinyl, indolyl, imidazolyl, oxazolyl, thiazolyl, pyrazolyl, pyridinyl, benzoxadiazolyl, benzothiadiazolyl, and naphthyl groups, all of which are optionally substituted. Substituted aryl groups include fully halogenated or semihalogenated aryl groups, such as aryl groups having one or more hydrogens replaced with one or more fluorine atoms, chlorine atoms, bromine atoms and/or iodine atoms. Substituted aryl groups include fully fluorinated or semifluorinated aryl groups, such as aryl groups having one or more hydrogens replaced with one or more fluorine atoms. Aryl groups include, but are not limited to, aromatic group-containing or heterocylic aromatic group-containing groups corresponding to any one of the following: benzene, naphthalene, naphthoquinone, diphenylmethane, fluorene, anthracene, anthraquinone, phenanthrene, tetracene, tetracenedione, pyridine, quinoline, isoquinoline, indoles, isoindole, pyrrole, imidazole, oxazole, thiazole, pyrazole, pyrazine, pyrimidine, purine, benzimidazole, furans, benzofuran, dibenzofuran, carbazole, acridine, acridone, phenanthridine, thiophene, benzothiophene, dibenzothiophene, xanthene, xanthone, flavone, coumarin, azulene or anthracycline. As used herein, a group corresponding to the groups listed above expressly includes an aromatic or heterocyclic aromatic group, including monovalent, divalent and polyvalent groups, of the aromatic and heterocyclic aromatic groups listed herein are provided in a covalently bonded configuration in the compounds of the invention at any suitable point of attachment. In embodiments, aryl groups contain between 5 and 30 carbon atoms. In embodiments, aryl groups contain one aromatic or heteroaromatic six-membered ring and one or more additional five- or six-membered aromatic or heteroaromatic ring. In embodiments, aryl groups contain between five and eighteen carbon atoms in the rings. Aryl groups optionally have one or more aromatic rings or heterocyclic aromatic rings having one or more electron donating groups, electron withdrawing groups and/or targeting ligands provided as substituents.
[0103] Arylalkyl groups are alkyl groups substituted with one or more aryl groups wherein the alkyl groups optionally carry additional substituents and the aryl groups are optionally substituted. Specific alkylaryl groups are phenyl-substituted alkyl groups, e.g., phenylmethyl groups. Alkylaryl groups are alternatively described as aryl groups substituted with one or more alkyl groups wherein the alkyl groups optionally carry additional substituents and the aryl groups are optionally substituted. Specific alkylaryl groups are alkyl-substituted phenyl groups such as methylphenyl. Substituted arylalkyl groups include fully halogenated or semihalogenated arylalkyl groups, such as arylalkyl groups having one or more alkyl and/or aryl groups having one or more hydrogens replaced with one or more fluorine atoms, chlorine atoms, bromine atoms and/or iodine atoms.
[0104] As to any of the groups described herein which contain one or more substituents, it is understood that such groups do not contain any substitution or substitution patterns which are sterically impractical and/or synthetically non-feasible. In addition, the compounds of this invention include all stereochemical isomers arising from the substitution of these compounds. Optional substitution of alkyl groups includes substitution with one or more alkenyl groups, aryl groups or both, wherein the alkenyl groups or aryl groups are optionally substituted. Optional substitution of alkenyl groups includes substitution with one or more alkyl groups, aryl groups, or both, wherein the alkyl groups or aryl groups are optionally substituted. Optional substitution of aryl groups includes substitution of the aryl ring with one or more alkyl groups, alkenyl groups, or both, wherein the alkyl groups or alkenyl groups are optionally substituted.
[0105] Optional substituents for any alkyl, alkenyl and aryl group includes substitution with one or more of the following substituents, among others:
halogen, including fluorine, chlorine, bromine or iodine;
pseudohalides, including -CN;
[0106] -COOR where R is a hydrogen or an alkyl group or an aryl group and more specifically where R is a methyl, ethyl, propyl, butyl, or phenyl group all of which groups are optionally substituted;
[0107] -COR where R is a hydrogen or an alkyl group or an aryl group and more specifically where R is a methyl, ethyl, propyl, butyl, or phenyl group all of which groups are optionally substituted;
[0108] -CON(R)2 where each R, independently of each other R, is a hydrogen or an alkyl group or an aryl group and more specifically where R is a methyl, ethyl, propyl, butyl, or phenyl group all of which groups are optionally substituted; and where R and R can form a ring which can contain one or more double bonds and can contain one or more additional carbon atoms;
[0109] -OCON(R)2 where each R, independently of each other R, is a hydrogen or an alkyl group or an aryl group and more specifically where R is a methyl, ethyl, propyl, butyl, or phenyl group all of which groups are optionally substituted; and where R and R can form a ring which can contain one or more double bonds and can contain one or more additional carbon atoms;
[0110] -N(R)2 where each R, independently of each other R, is a hydrogen, or an alkyl group, or an acyl group or an aryl group and more specifically where R is a methyl, ethyl, propyl, butyl, phenyl or acetyl group, all of which are optionally substituted; and where R and R can form a ring which can contain one or more double bonds and can contain one or more additional carbon atoms; [0111] -SR, where R is hydrogen or an alkyl group or an aryl group and more specifically where R is hydrogen, methyl, ethyl, propyl, butyl, or a phenyl group, which are optionally substituted;
-S02R, or -SOR where R is an alkyl group or an aryl group and more specifically where R is a methyl, ethyl, propyl, butyl, or phenyl group, all of which are optionally substituted;
[0112] -OCOOR where R is an alkyl group or an aryl group;
[0113] -S02N(R)2 where each R, independently of each other R, is a hydrogen, or an alkyl group, or an aryl group all of which are optionally substituted and wherein R and R can form a ring which can contain one or more double bonds and can contain one or more additional carbon atoms;
[0114] -OR where R is H, an alkyl group, an aryl group, or an acyl group all of which are optionally substituted. In a particular example R can be an acyl yielding -OCOR" where R" is a hydrogen or an alkyl group or an aryl group and more specifically where R" is methyl, ethyl, propyl, butyl, or phenyl groups all of which groups are optionally substituted.
[0115] Specific substituted alkyl groups include haloalkyl groups, particularly trihalomethyl groups and specifically trifluoromethyl groups. Specific substituted aryl groups include mono-, di- , tri, tetra- and pentahalo-substituted phenyl groups; mono-, di-, tri-, tetra-, penta-, hexa-, and hepta-halo-substituted naphthalene groups; 3- or 4-halo-substituted phenyl groups, 3- or 4-alkyl- substituted phenyl groups, 3- or 4-alkoxy-substituted phenyl groups, 3- or 4-RCO-substituted phenyl, 5- or 6-halo-substituted naphthalene groups. More specifically, substituted aryl groups include acetylphenyl groups, particularly 4-acetylphenyl groups; fluorophenyl groups, particularly
3- fluorophenyl and 4-fluorophenyl groups; chlorophenyl groups, particularly 3-chlorophenyl and
4- chlorophenyl groups; methylphenyl groups, particularly 4-methylphenyl groups; and methoxyphenyl groups, particularly 4-methoxyphenyl groups.
DETAILED DESCRIPTION
[0116] The following definitions and methods are provided to better define the invention and to guide those of ordinary skill in the art in the practice of the invention.
[0117] Referring to the drawings, like numerals indicate like elements and the same number appearing in more than one drawing refers to the same element. Unless otherwise noted, the terms and phrases used herein have their art-recognized meaning, which can be found by reference to standard texts, journal references and contexts known to those skilled in the relevant art.
[0118] The term "inflammation" generally refers to a biological response of tissues to harmful stimuli, such as pathogens, damaged cells, irritants, etc. Inflammation can be either acute or chronic. Acute inflammation is an initial response of the body to harmful stimuli and can be achieved by the increased movement of plasma and leukocytes from the blood into injured tissues. An inflammatory response can involve the local vascular system, the immune system, and/or various cells within the injured tissue. Prolonged inflammation, referred to as chronic inflammation, can lead to a progressive shift in the type of cells which are present at the site of inflammation can be characterized by simultaneous destruction and healing of the tissue from the inflammatory process.
[0119] The term "amino acid" comprises naturally occurring amino acids as well as non- naturally occurring amino acids, including amino acid analogs and derivatives. One skilled in the art will recognize that reference herein to an amino acid includes, for example, naturally occurring proteogenic L-amino acids; D-amino acids; chemically modified amino acids such as amino acid analogs and derivatives; naturally occurring non-proteogenic amino acids, and chemically synthesized compounds having properties known in the art to be characteristic of amino acids.
[0120] The term "nucleic acid" as used herein generally refers to a molecule or strand of DNA, RNA, or derivatives or analogs thereof including one or more nucleobases. Nucleobases comprise purine or pyrimidine bases typically found in DNA or RNA (e.g., adenine, guanine, thymine, cytosine, and/or uracil). The term "nucleic acid" also comprises oligonucleotides and polynucleotides. Nucleic acids may be single-stranded molecules, or they may be double-, triple- or quadruple-stranded molecules that may comprise one or more complementary strands of a particular molecule. "Nucleic acid" includes artificial nucleic acids including peptide nucleic acids, morpholino nucleic acids, glycol nucleic acids and threose nucleic acids. Artificial nucleic acids may be capable of nucleic acid hybridization.
[0121] As used herein, "sequence" means the linear order in which monomers occur in a polymer, the order of amino acids in a polypeptide or the order of nucleotides in a polynucleotide for example.
[0122] The terms "peptide" and "polypeptide" are used synonymously in the present description, and refer to a class of compounds comprising of amino acid residues chemically bonded together by amide bonds (or peptide bonds), regardless of length, functionality, environment, or associated molecule(s). Peptides and polypeptides are polymeric compounds comprising at least two amino acid residues or modified amino acid residues. Modifications can be naturally occurring or non-naturally occurring, such as modifications generated by chemical synthesis. Modifications to amino acids in peptides include, but are not limited to, phosphorylation, glycosylation, lipidation, prenylation, sulfonation, hydroxylation, acetylation, methionine oxidation, alkylation, acylation, carbamylation, iodination and the addition of cofactors. Peptides include proteins and further include compositions generated by degradation of proteins, for example by proteolyic digestion. Peptides and polypeptides can be generated by substantially complete digestion or by partial digestion of proteins. Polypeptides comprising 2 to 100 amino acid units, optionally for some embodiments 2 to 50 amino acid units and, optionally for some embodiments 2 to 20 amino acid units can be used as polypeptide targeting ligands in the invention, for example, where the polypepetide preferentially binds to proteins, peptides or other biomolecules expressed, or otherwise generated by, a target tissue, such as a tumor, precancerous tissue, site of inflammation or other lesion. Typically, the polypeptide is at least four amino acid residues in length and can range up to a full-length protein.
[0123] "Protein" refers to a class of compounds comprising one or more polypeptide chains and/or modified polypeptide chains. Proteins can be modified by naturally occurring processes such as post-translational modifications or co-translational modifications. Exemplary post- translational modifications or co-translational modifications include, but are not limited to, phosphorylation, glycosylation, lipidation, prenylation, sulfonation, hydroxylation, acetylation, methionine oxidation, the addition of cofactors, proteolysis, and assembly of proteins into macromolecular complexes. Modification of proteins can also include non-naturally occurring derivatives, analogues and functional mimetics generated by chemical synthesis. Exemplary derivatives include chemical modifications such as alkylation, acylation, carbamylation, iodination or any modification that derivatizes the protein.
[0124] As used herein, "polynucleotide" and "oligonucleotide" are used interchangeably and refer to a class of compounds composed of nucleic acid residues chemically bonded together. The invention provides optical agents having an oligonucleotide or polynucleotide targeting ligand which comprises a plurality of nucleic acid residues, such as DNA or RNA residues, and/or modified nucleic acid residues that preferentially binds to proteins, peptides or other biomolecules expressed, or otherwise generated by, a target tissue, such as a tumor, precancerous tissue, site of inflammation or other lesion. Modifications to nucleic acid residues can be naturally occurring or non-naturally occurring, such as modifications generated by chemical synthesis. Oligo- or poly-nucleotide targeting ligands include, for example, oligo- or poly-nucleotides comprising 2 to 100 nucleic acid units, optionally for some embodiments 2 to 50 nucleic acid units and, optionally for some embodiments 2 to 20 nucleic acid units, and optionally for some embodiments 2 to 10 nucleic acid units. Polypeptide and oligonucleotide include a polymer of at least two nucleotides joined together by phosphodiester bonds and may consist of either ribonucleotides or deoxyribonucleotides.
[0125] The term "aptamer" refers to an oligo- or poly-nucleotide or polypeptide that binds to, or otherwise selectively or preferentially associates with, a specific target molecule. For example, the invention provides optical agents having an aptamer targeting ligand that preferentially binds to proteins, peptides or other biomolecules expressed, or otherwise generated by, a target tissue, such as a tumor, precancerous tissue, site of inflammation or other lesion.
[0126] "Peptidomimetic" refers to a molecule having activity, including biological activity, that resembles that of a polypeptide or is substantially the same as a polypeptide. Morphine, for example, is a peptidomimetic of endorphin peptide. In some embodiments, a peptidomimetic is a small protein-like polymer designed to mimic the functionality of a peptide. Peptidomimetics useful as targeting ligands for some compounds of the invention in the present invention include peptoids and β-peptides. The composition and biological activity of peptidomimetics and use of peptidomimetics in targeted diagnostics and therapeutics are further described in the following references: (1 ) A. Giannis and T. Kolter, Peptidomimetics for Receptor Ligands - Discovery, Development, and Medical Perspectives, Angewandte Chemie International Edition In English, vol. 32, 1993, pg. 1244-1267; (3) Peptidomimetics, Accounts of Chemical Research, Vol. 41 , No. 10, October 208, 1231-1232, by Wu and Gellman; and (3) Patch, J.A. et al., Versatile oligo(N-substituted)glycines: The many roles of peptoids in drug discovery., Pseudo-Peptides in Drug Discovery 2004, 1-31 P.E. Nielsen.
[0127] As used herein, "attaching moiety" refers to a component provided to attach an optical dye, such as a pyrazine, cyanine, azulene, azaazulene or indocyanine group, directly or indirectly to central sulfone hydroxamic acid or hydroxamate backbone in compounds of the invention. In some embodiments, L1, L2, W1 and/or W2 in formulas (FX1 ) - (FX113) are an attaching moieties.
[0128] As used herein, an "electron withdrawing group" (abbreviated as "EWG") refers to a chemical group that draws electrons or electron density from a center, such as optical dye component of compounds of the invention, for example a pyrazine, cyanine, azulene, azaazulene or indocyanine group. In some embodiments, the electron withdrawing group(s) are independently selected from cyano (-CN), carbonyl (-CO), carboxylate (-C02Ra), halo (-F, -CI, -Br, -I, -At), carbamate (-CONRbRc), acyl (-CORd), nitro (-N02), sulfinyl (-SORe), sulfonyl (- S02R , -SO2OR9, and -POsR^, wherein in the context of this description, Ra to R' are independently selected to enhance biological and/or physiochemical properties of the optical agents of the invention. In some instances, Ra to R' are independently selected from any one of a hydrogen atom, an anionic functional group (e.g., carboxylate, sulfonate, sulfate, phosphonate or phosphate) and a hydrophilic functional group (e.g., hydroxyl, carboxyl, sulfonyl, sulfonato or phosphonato). In other instances, Ra to R' are independently selected from hydrogen, C1-10 alkyl, aryl, heteroaryl, -(CH2)nOH, -(CH2)nC02H, -(CH2)nS03H, -(CH2)nS03- -(CH2)nOS03H, - (CH2)nOS03- -(CH2)nNHS03H, -(CH2)nNHS03- -(CH2)nP03H2, -(CH2)nP03l-r, -(CH2)nP03 =, - (CH2)nOP03H2, -(CH2)nOP03H" and -(CH2)nOP03, wherein n is an integer from 1 to 10. In one example of this embodiment, the EWG(s) are independently selected from is -CN, -C02R40, - S02OR43, -CONR52R53, -COR54, -N02, -SOR41 ; -S02R55, -POsR^R45, halo, C C6 acyl, trihalomethyl, or -S02NR56R57, wherein each of R40 to R59 is independently H or d - C10 alkyl. In an embodiment, an EWG is located at the terminus of a substituent arm of a an optical dye of compounds of formulas (FX1 ) - (FX113), such pyrazine, cyanine, azulene, azaazulene or indocyanine group of the compounds of compounds of formulas (FX1 ) - (FX113).
[0129] As used herein, an "electron donating group" (abbreviated as "EDG") refers to a chemical group that releases electrons or electron density to a center, such as optical dye component of compounds of the invention, for example a pyrazine, cyanine, azulene, azaazulene or indocyanine group. In some embodiments, the electron donating group(s) are independently selected from C1-C10 alkyl, C5-C10 aryl, -(CH2)zOH, -ORj, -SRk, -NR'Rm, - N(Rn)COR°, and -P(R ), wherein in the context of this description, Rj to R are independently selected to enhance biological and/or physiochemical properties of the optical agents of the invention and wherein z is selected from the range of 1 to 10. In some instances, Rj to R are independently selected from any one of a hydrogen atom, an anionic functional group (e.g., carboxylate, sulfonate, sulfate, phosphonate or phosphate) and a hydrophilic functional group (e.g., hydroxyl, carboxyl, sulfonyl, sulfonato or phosphonato). In other instances, Rj to R are independently selected from hydrogen, C1-10 alkyl, aryl, heteroaryl, -(CH2)nOH, -(CH2)zC02H, - (CH2)zS03H, -(CH2)nS03 ", -(CH2)zOS03H, -(CH2)zOS03 ", -(CH2)zNHS03H, -(CH2)zNHS03 ", - (CH2)zP03H2, -(CH2)zP03H", -(CH2)ZP03 =, -(CH2)ZOP03H2, -(CH2)ZOP03H_ and -(CH2)zOP03 = where z is an integer from 1 to 10. In one example of this embodiment, the EDG(s) are independently C C6 alkyl, C3-C6 cycloalkyl -OR46, -SR47, -NR48R49, or -NR50COR51 , wherein each of R46 to R51 is independently H or C-i - C10 alkyl. In an embodiment, an EDG is located at the terminus of a substituent arm of a substituent arm of a an optical dye of compounds of formulas (FX1 ) - (FX1 13), such pyrazine, cyanine, azulene, azaazulene or indocyanine group of the compounds of compounds of formulas (FX1 ) - (FX1 13).
[0130] In embodiments, two substituents, such as EDG and EWG substituents, on a compound of the invention can act in what is known as a "push-pull" arrangement. In embodiments of the "push-pull" arrangement, the electron density of the compound or a portion thereof, such as an aryl or heteroaryl group, is polarized due in part to the location of an EWG and EDG on the compound. In embodiments of the "push-pull' arrangement, an EWG is positioned at a terminus of a substituent arm of the structure and an EDG is positioned at a terminus of a different substituent arm of the structure. In embodiments of the "push-pull" arrangement, an EWG is positioned at one end of a π bond and an EDG is positioned at the other end of a π bond. In an embodiment, an EWG is positioned para- to an EDG in a six- membered ring structure. In an embodiment, an EWG is positioned trans- to an EDG in an alkylene structure. In some embodiments, compounds having the "push-pull" arrangement exhibit a shift in the optical absorbance and emission spectrum as compared to compounds not having the "push-pull" arrangement.
[0131] "Optical agent" generally refers to compounds, compositions, preparations, and/or formulations that absorb, emit, or scatter electromagnetic radiation of wavelength generally in the range of 350 - 1300 nanometers, within a biologically relevant environment or condition. In some embodiments, optical agents of the invention, when excited by electromagnetic radiation, undergo emission via fluorescence or phosphorescence pathways. These pathways are useful for diagnostic imaging, visualization, or organ function monitoring. Compounds belonging to this class are commonly referred to as "optical imaging agents" or "optical contrast agents." In some other embodiments, optical agents of the invention absorb electromagnetic radiation and undergo photochemical reactions such as photofragmentation of one or more photolabile bonds to generate reactive species such as nitrenes, carbene, free radicals, ions, excited species, etc. This process is useful for a wide range of phototherapy applications, for example in the treatment of tumors or other lesions. Compounds belonging to this class are commonly referred to as "photosensitizers." The term "photosensitizer" refers to a phototherapeutic agent or a component thereof providing for photoactivation, for example, photoactivation resulting in generation of reactive species that locally kill, injure, inactivate or otherwise degrade cells (e.g., cancer cells, tumor cells, non-cancer cells, etc.). Photosensitizers of some embodiments undergo photoactivation that initiates bond cleavage reactions, such as photolysis and/or nitrogen extrusion reactions, thereby generating reactive species capable of causing localized cell death or injury. Optical agents include Type 1 and Type 2 phototherapeutic agents. Optical agents include, but are not limited to, phototherapeutic agents (Type 1 and 2), photosensitizers, imaging agents, dyes, detectable agents, photosensitizer agents, photoactivators, and photoreactive agents; and conjugates, complexes, and derivatives thereof.
[0132] As used herein, a "chromophore" is a compound or functional group of a compound that results in absorption of electromagnetic radiation, preferably for some applications electromagnetic radiation having wavelengths in the UV (e.g. 200 nm to 350 nm) or visible (e.g. 350 nm to 750 nm) of the electromagnetic spectrum.
[0133] As used herein, a "fluorophore" is a compound or functional group of a compound that results in absorption of electromagnetic radiation and subsequent fluorescence. Preferably for some applications incorporation of a fluorophore results in compounds of the invention that absorb electromagnetic radiation and generate fluorescence having wavelengths in the UV (e.g. 200 nm to 350 nm) or visible (e.g. 350 nm to 750 nm) of the electromagnetic spectrum. In some embodiment, incorporation of a fluorophore results in compounds having an appreciable quantum yield for fluorescence, such as a quantum yield over the range of 0.001 to 1 , 0.01 to 1 , optionally 0.1 to 1. Optical agents of the present invention can contain fluorophores. Fluorophores can be functional groups in a molecule which absorb electromagnetic radiation of first specific wavelengths and re-emit energy at second specific wavelengths. The amount and wavelengths of the emitted electromagnetic radiation depend on both the fluorophore and the chemical environment of the fluorophore. The term "fluorophore" may be abbreviated throughout the present description as "FL". In aspects of the invention, fluorophores emit energy in the visible (e.g. 350 nm to 750 nm) and NIR regions (e.g., 750 - 1300nm) of the electromagnetic spectrum.
[0134] As used herein, the term "luminescence" refers to the emission of electromagnetic radiation from excited electronic states of atoms or molecules. Luminescence generally refers to electromagnetic radiation emission, such as photoluminescence, chemiluminescence, and electrochemiluminescence, among others. In photoluminescence, including fluorescence and phosphorescence, the excited electronic state is created by the absorption of electromagnetic radiation. Luminescence detection involves detection of one or more properties of the luminescence or associated luminescence process. These properties can include intensity, excitation and/or emission spectrum, polarization, lifetime, and energy transfer, among others. These properties can also include time-independent (steady-state) and/or time-dependent (time- resolved) properties of the luminescence. Representative luminescence techniques include fluorescence intensity (FLINT), fluorescence polarization (FP), fluorescence resonance energy transfer (FRET), fluorescence lifetime (FLT), total internal reflection fluorescence (TIRF), fluorescence correlation spectroscopy (FCS), fluorescence recovery after photobleaching (FRAP), and bioluminescence resonance energy transfer (BRET), among others. By way of example, when an optical agent is used in the present invention, it is desirable that the wavelength of radiation be non-ionizing and be such that it excites the optical agent. This excitation can cause a bond of the molecule to break and can lead to creation of one or more appropriate radical(s). This excitation can also cause the molecule to emit part of the absorbed energy at a different wavelength. Such emission can be detected using fluorometric techniques as described above. One skilled in the art can readily determine the most appropriate treatment and optional detection technique based, at least in part, on the specific phototherapeutic agent(s) administered and/or the particular use (e.g., tissue to be treated).
[0135] Optical condition" refers to one or more of the following: the fluorescence quantum yield, fluorescence intensity, fluorescence excitation wavelength, wavelength distribution or spectrum, emission wavelength, wavelength distribution or spectrum, Stokes shift, color, reflectance, phosphorescence, chemiluminescence, scattering, and/or other observable and/or measurable spectral property or phenomenon.
[0136] "Phototherapy procedure" refers to a therapeutic procedure involving administration of a phototherapeutic agent to a patient followed by subsequent excitation by exposure to applied electromagnetic radiation, such as electromagnetic radiation having wavelengths in the visible and/or near IR region of the electromagnetic spectrum. Such wavelengths can be in the range of 350 - 1300 nanometers, so as to generate a therapeutically effective amount of excited phototherapeutic agent. Phototherapy includes, but is not limited to, photodynamic therapy. As used herein, "phototherapy" includes procedures involving administration of Type 1 and/or Type 2 phototherapeutic agents, optionally further including administration of one or more additional therapeutic agents.
[0137] A detectable optical signal may be, for example, an observable change in absorbance, reflectance, phosphorescence, chemiluminescence, scattering, or other spectral property.
[0138] As used herein, "tumor-specific agent" refers to a compound or composition, such as an optical agent, that preferentially accumulates in a tumor at a higher level than normal tissue regardless of the particular mechanism of uptake in the tumors, for example, receptor mediated or enhanced permeability and retention (EPR). Optical agents of the invention include tumor- specific agents, including tumor specific phototherapy agents, for example having a targeting ligand providing specificity in the administration, delivery and/or binding to tumor tissue. [0139] As used herein, "targeting ligand" (abbreviated as Bm) refers to a chemical group and/or substituent having functionality for targeting a compound of the invention to an anatomical and/or physiological site of a patient, such as a selected cell, tissue or organ. For some embodiments, a targeting ligand is characterized as a ligand that selectively or preferentially binds to a specific biological site(s) (e.g., enzymes, receptors, etc.) and/or biological surface(s) (e.g., membranes, fibrous networks, etc.).
[0140] "Target tissue" refers to tissue of a subject to which an optical agent is administered or otherwise contacted, for example during a biomedical procedure such as an optical imaging, phototherapy, monitoring or visualization procedure. Target tissues can be contacted with an optical agent of the invention under in vivo conditions in vitro conditions or ex vivo conditions. Target tissues in some embodiments include cancerous tissue, cancer cells, precancerous tissue, a tumor, a lesion, a site of inflammation, or vasculature tissue. In some embodiments, a target tissue includes a melanoma cell, a breast lesion, a prostate lesion, a lung cancer cell, a colorectal cancer cell, an atherosclerotic plaque, a brain lesion, a blood vessel lesion, a lung lesion, a heart lesion, a throat lesion, an ear lesion, a rectal lesion, a bladder lesion, a stomach lesion, an intestinal lesion, an esophagus lesion, a liver lesion, a pancreatic lesion, and a solid tumor. Target tissue in some embodiments refers to a selected organ of the subject or component thereof, such as lung, heart, brain, stomach, liver, kidneys, gallbladder, pancreas, intestines, rectum, skin, colon, prostate, ovaries, breast, bladder, blood vessel, throat, ear, or esophagus.
[0141] Methods of this invention comprise the step of administering an "effective amount" of the present diagnostic and therapeutic compositions, formulations and preparations containing the present compounds or compositions, to diagnose, image, monitor, evaluate, treat, reduce, alleviate, ameliorate or regulate a biological condition and/or disease state in a patient. The term "effective amount," as used herein, refers to the amount of the diagnostic and therapeutic formulation, that, when administered to the individual is effective to diagnose, image, monitor, evaluate, treat, reduce alleviate, ameliorate or regulate a biological condition and/or disease state. As is understood in the art, an effective amount of a given composition or formulation will depend at least in part upon the mode of administration (e.g. intravenous, oral, topical administration), any carrier or vehicle employed, and the specific individual to whom the formulation is to be administered (age, weight, condition, sex, etc.). The dosage requirements needed to achieve the "effective amount" vary with the particular formulations employed, the route of administration, and clinical objectives. Based on the results obtained in standard pharmacological test procedures, projected daily dosages of active compound or composition can be determined as is understood in the art.
[0142] In an embodiment, an effective amount of a compound or composition of the invention is a therapeutically effective amount. As used herein, the phrase "therapeutically effective" qualifies the amount of compound or composition administered in the therapy. This amount achieves the goal of ameliorating, suppressing, eradicating, preventing, reducing the risk of, or delaying the onset of a targeted condition. In an embodiment, an effective amount of a compound or composition of the invention is a diagnostically effective amount. As used herein, the phrase "diagnostically effective" qualifies the amount of compound or composition administered in diagnosis, for example of a disease state or other pathological condition. The amount achieves the goal of being detectable while avoiding adverse side effects found with higher doses. In an embodiment, an active ingredient or other component is included in a therapeutically acceptable amount. In an embodiment, an active ingredient or other component is included in a diagnostically acceptable amount.
[0143] It is contemplated that the compounds and pharmaceutically acceptable salts of the invention can be used as part of a combination. The term "combination" means the administration of two or more compounds directed to a target condition. The treatments of the combination generally can be co-administered in a simultaneous manner. Two compounds can be co-administered as, for example: (a) a single formulation (e.g., a single capsule) having a fixed ratio of active ingredients; or (b) multiple, separate formulations (e.g., multiple capsules) for each compound. The treatments of the combination can alternatively (or additionally) be administered at different times.
[0144] In certain embodiments, the invention encompasses administering optical agents useful in the invention to a patient or subject. A "patient" or "subject", used equivalently herein, refers to an animal. In particular, an animal refers to a mammal, preferably a human. The subject can either: (1 ) have a condition able to be monitored, diagnosed, prevented and/or treated by administration of an optical agent of the invention; or (2) is susceptible to a condition that is able to be monitored, diagnosed, prevented and/or treated by administering an optical agent of the invention.
[0145] When used herein, the terms "diagnosis", "diagnostic" and other root word derivatives are as understood in the art and are further intended to include a general monitoring, characterizing and/or identifying a state of health or disease. The term is meant to encompass the concept of prognosis. For example, the diagnosis of cancer can include an initial determination and/or one or more subsequent assessments regardless of the outcome of a previous finding. The term does not necessarily imply a defined level of certainty regarding the prediction of a particular status or outcome.
[0146] As defined herein, "administering" means that a compound or formulation thereof of the invention, such as an optical agent, is provided to a patient or subject, for example in a therapeutically effective amount. The invention includes methods for a biomedical procedure wherein a therapeutically or diagnostically effective amount of a compound having any one of formulas (FX1 ) - (FX113) is administered to a patient in need of treatment, for example to a patient undergoing treatment for a diagnosed diseased state including cancer and vascular diseases. Administering can be carried out by a range of techniques known in the art including parenteral administration including intravenous, intraperitoneal or subcutaneous injection or infusion, oral administration, topical or transdermal absorption through the skin, or by inhalation, for example. The chosen route of administration may depend on such factors as solubility of the compound or composition, location of targeted condition, and other factors which are within the knowledge of one having ordinary skill in the relevant art.
[0147] "Topical administration" includes the use of transdermal administration, such as transdermal patches or iontophoresis devices.
[0148] "Parenteral administration" includes subcutaneous injections, intravenous injections, intraarterial injections, intraorbital injections, intracapsular injections, intraspinal injections, intraperitoneal injections, intramuscular injections, intrasternal injections, and infusion. Dosage forms suitable for parenteral administration include solutions, suspensions, dispersions, emulsions, and any other dosage form that can be administered parenterally.
[0149] As used herein, the term "controlled-release component" refers to an agent that facilitates the controlled-release of a compound including, but not limited to, polymers, polymer matrices, gels, permeable membranes, liposomes, microspheres, or the like, or any combination thereof. Methods for producing compounds in combination with controlled-release components are known to those of skill in the art.
[0150] As used herein, the term "pharmaceutically acceptable" means approved by a regulatory agency of an appropriate federal or state government; or listed in the U.S. Pharmacopoeia or other generally recognized pharmacopoeia for use in animals, and more particularly in humans; or does not impart significant deleterious or undesirable effect on a subject to whom it is administered and in the context in which it is administered.
[0151] As will be clear to those of ordinary skill in the art, the groups and structures described herein as portions of the compounds of the invention may be defined as if they are separate valence-satisfied chemical structures. It is intended that when a group is described or shown as being a substituent of another group, that the group be viewed as having a valency to allow this binding to occur.
[0152] The invention is further detailed in the following Examples, which are offered by way of illustration and are not intended to limit the scope of the invention in any manner.
Example 1 : Optically Functional Matrix Metalloproteinase Inhibitors
[0153] Matrix metalloproteinases (MMPs) are zinc-dependent enzymes responsible for the remodeling and degradation of components of the extracellular matrix. The up-regulation of MMPs has been implicated in numerous diseases including osteoarthritis and cancer. A key therapeutic target for MMPs is cancer, as MMPs appear important for tumor growth and metastasis processes, and inhibition of MMP-9 in particular appears able to block metastasis. Until recently, however, clinical trials with MMP inhibitors for advanced cancer(s) have not been successful in demonstrating efficacy. Bramhall has now reported a placebo-controlled double blind study reporting success in treating cancer with an MMP inhibitor in a study treating gastric cancer patients with the broad spectrum inhibitor marimastat. [Bramhall, S.R et al.; Marimastat as maintenance therapy for patients with advanced gastric cancer: A randomized trial. Br. J. Cancer, 2002, 86, 1864-1870]. A survival benefit has also been recently demonstrated in glioblastoma multiform patients on marimastat in combination with temozolomide, providing additional support that MMP inhibitors can improve the outcome of cancer patients. [Groves, M.D. et al., Phase II trial of temozolomide plus the matrix metalloproteinase inhibitor, marimastat, in recurrent and progressive glioblastoma multiform, J. Clin. Oncol. 2002, 20, 1383 - 1388]. Marimastat afforded a survival rate similar to Gemcitabine in patients with unresectable pancreatic cancer. [Bramhall, S.R. et al., Marimastat as first-line therapy for patients with unresectable pancreatic cancer: a randomized trial, J. Clin. Oncol., 2001 , 19, 3447-3455]. Thus, the proof-of principle for efficacy in treating human cancers with MMP inhibitors appears to have been clinically demonstrated.
[0154] MMPs have been shown to be up-regulated; most notably in osteoarthritis and in cardiovascular disease as there is significant up-regulation of MMP-2 and MMP-13 immediately after an ischemic event. There is also evidence that treatment with an MMP inhibitor immediately after an ischemic event for a defined acute period has significant beneficial effects on left ventricular re-modeling. This therapeutic approach may have potential in an intensive care unit environment. MMP-2 and MMP-9 have recently been implicated in neuropathic pain states; both acute and chronic. A way to image the neuropathic pain pathway via imaging tissue expressing MMPs would also be beneficial as there are many aspects of the fundamental biology that are poorly understood
[0155] Compounds of the present invention include optical agents providing target-specific small molecule optical imaging agents that inhibit the biological activity of various MMPs. The present compounds are useful for optical evaluation and monitoring of biological materials where MMPs play a role in biological pathways related to the above mentioned diseases and conditions. In addition, the invention includes methods of using optically functional MMP inhibitors to improve patient outcomes by enabling imaging and visualization techniques, optionally in combination with other anticancer or cardiovascular (or other disease where coadministration of above mentioned MMP Inhibitor would be beneficial) drug treatment regimens.
[0156] Optical agents of some embodiments comprise a sulfone hydroxamic acid or hydroxamate backbone and a fluorescent moiety having substituents providing for optical functionality useful for optical imaging, therapy and/or detection. Preferably, the sulfone hydroxamic acid or hydroxamate backbone and a fluorescent moiety are covalently linked such that the molecular recognition functionality for MMPs is preserved. In some embodiments, for example, an optical dye comprising a fluorescent moiety is provided in the solvent exposed portion of the MMP binding pocket and in other embodiments an optical dye comprising a fluorescent moiety is provided in the S1 ' MMP binding pocket. Examples of optically functional matrix metalloproteinase inhibitors useful for detection, imaging and diagnosis of target tissue expressing MMPs are provided in Table 1 . In the formulas provided in Table 1 , R3, R6, R16, R; and a are as set forth and described in the context of formulas (FX1 ) - (FX113).
Table 1 : Optically Functional Matrix Metalloproteinase Inhibitors
Figure imgf000071_0001
No. COMPOUND
Figure imgf000072_0001
[0157] Compounds having formulas shown in nos. 1 , 2, and 3 in Table 1 incorporate the fluorescent moiety in a solvent exposed portion of the binding pocket and compounds having formulas shown in nos. 4, 5, 6 and 7 in Table 1 incorporate the fluorescent moiety in the SV binding pocket. The position of the optical dye and linking configuration to the backbone may be selected to control the shift and/or magnitude of the fluorescence of the fluorescent moiety core. The compound having the formula shown in no. 4 of Table 1 is also expected to have activity against a related metalloproteinase called TNF-a Converting Enzyme (TACE) which is implicated in the cartilage catabolism pathway and is a specific target of interest within the pharmaceutical industry.
Example 2: Inhibition of MMPs
[0158] To evaluate their inhibitory activity for MMPs, a number of optically functional MMP inhibitors were synthesized and characterized with respect to their percent inhibition of specific binding or activity maximal inhibitory concentration (IC50) with respect to MMP-2 and MMP-9. Compounds evaluated were also characterized with respect to certain optical properties including maximum absorption wavelength (λ^)) and maximum emission wavelength (λ(βΧ)) in visible and infrared regions.
[0159] Assays used for determining inhibitory activity are described in: (1 ) Knight CG, Willenbrock F and Murphy G (1992) "A novel coumarin-labelled peptide for sensitive continuous assays of the matrix metalloproteinases," FEBS Lett. 296(3): 263-266; and (2) Olson MW, Gervasi DC, Mobashery S and Fridman R (1997), "Kinetic analysis of the binding of human matrix metalloproteinase-2 and -9 to tissue inhibitor of metalloproteinase (TIMP)-1 and TIMP-2," Biol Chem. 272 (47): 29975 - 29983. Tables 2 and 3 summarize the experimental conditions employed in the assays for determining inhibitory activity for MMP-2 and MMP-9. Table 4 summarizes the composition and optical properties of the compounds evaluated.
[0160] Biochemical assay results are presented as the percent inhibition of specific binding or activity. All other results are expressed in terms of that assay's quantitation method. For primary assays, only the lowest concentration with a significant response judged by the assays' criteria, is shown. Where applicable, either the secondary assay results with the lowest
dose/concentration meeting the significance criteria or, if inactive, the highest
dose/concentration that did not meet the significance criteria is shown. Unless otherwise requested, primary screening in duplicate with quantitative data (e.g., IC50 ± SEM, Ki ± SEM and nH) are shown where applicable for individual requested assays. In screening packages, primary screening in duplicate with semi-quantitative data (e.g., estimated IC50, Ki and nH) are shown where applicable (concentration range of 4 log units); available secondary functional assays are carried out (30 mM) and MEC or MIC determined only if active in primary assays >50% at 1 log unit below initial test concentration. Table 2: Experimental conditions for MMP-2 inhibition assay
Source: Human recombinant
Substrate: 4 μΜ Mca-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2
Vehicle: 1 % DMSO
Pre-lncubation Time/Temp: 60 minutes @ 37°C
Incubation Time/Temp: 2 hours @ 37°C
Incubation Buffer: 50 mM MOPS, pH 7.2, 10 mM
CaCI2, 10 μΜ ZnCI2, 0.05% Brij 35
Quantitation Method: Spectrofluorimetric quantitation of Mca-Pro-Leu-Gly
Significance Criteria: ≥ 50% of max stimulation or inhibition
Table 3: Experimental conditions for MMP-9 inhibition assay
Source: Human recombinant
Substrate: 4 μΜ Mca-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2
Vehicle: 1 % DMSO
Pre-lncubation Time/Temp: 60 minutes @ 37°C
Incubation Time/Temp: 2 hours @ 37°C
Incubation Buffer: 50 mM MOPS, pH 7.2, 10 mM
CaCI2, 10 μΜ ZnCI2, 0.05% Brij 35
Quantitation Method: Spectrofluorimetric quantitation of Mca-Pro-Leu-Gly
Significance Criteria: ≥ 50% of max stimulation or inhibition
Table 4: MMP inhibitors evaluated
No. COMPOUND OPTICAL PROPERTIES
λιηαχίε δ) - 400 nm ιηαχ(βχ) - 500 nm
Figure imgf000074_0001
Figure imgf000075_0001
[0161] Table 5 summarizes percentage inhibition values obtained for compounds having formula (FX105) - (FX109). Figures 1 - 5 provide response curves obtained for some of the compounds evaluated. Closed circles in Figures 1-5 represent percentage inhibition values for compounds of the present invention. The closed squares in Figures 1-5 represent percentage inhibition values for control measurements for TIMP-2 (Tissue Inhibitor Metalloprotease-2) which is a natural regulator of MMPs. Figure 1 provides the response curve for inhibition of MMP-2 by compound (FX107). Figure 2 provides the response curve for inhibition of MMP-9 by compound (FX107). Figure 3 provides the response curve for inhibition of MMP-2 by compound (FX108). Figure 4 provides the response curve for inhibition of MMP-2 by compound (FX105). Figure 5 provides the response curve for inhibition of MMP-9 by compound (FX105). In Figures 1 -5, percentage inhibition (%) is plotted as a function of concentration (μηι) of the optically functional MMP inhibitor compound.
Table 5. Percentage Inhibition Values for MMP-2 and MMP-9
Figure imgf000076_0001
[0162] Table 6 summarizes experimental results in connection with characterization of the inhibitor activity of the compounds evaluated. Parameters provided in Table 6 include percentage inhibition and half maximal inhibitory concentration (IC50) in connection with selective inhibition of MMP-2 and MMP-9. The optically functional MMP inhibitors exhibit good activity (e.g., IC50 < 10 nM) with respect to MMP-2 and MMP-9. It is important to note that comparable activity was observed for MMP inhibitors having pyrazine and indocyanine groups, suggesting that the presence of the optical dye component does not appreciable impact selective binding. In addition, comparable activity was observed for MMP inhibitors with and without a short spacer positioned between the hydroxamic acid backbone and the optical dye components.
Table 6: Experimental results for MMP inhibition
Figure imgf000077_0001
Example 3: Methods of Using Optically Functional MMP Inhibitors
[0163] In an embodiment, the present invention provides methods of using optically functional MMP inhibitors that selectively bind to MMP enzymes. Such optically functional MMP inhibitors can be used for detecting, monitoring and imaging target tissue and cells, and in particular cancer cells, that express MMPs at higher levels than normal cells. In addition, these optical agents provide MMP inhibitors useful for treatment of pathological conditions associated with MMP enzymes, such as cancer and inflammation. Without being bound by a particular theory, it is understood that the sulfone hydroxamic acid or hydroxamate backbone of the optically functional MMP inhibitors serve to bind selectively the MMPs, whereas the fluorescent moiety functions as an optical tag or probe that enables detection or imaging of tissue and cells that express MMPs. In general, molecules absorbing, emitting, or scattering in the visible, NIR, or long-wavelength (UV-A, > 300 nm) region of the electromagnetic spectrum are useful for optical detection and/or imaging. The high sensitivity associated with fluorescence permits detection without the negative effects of radioactivity or ionizing radiation. Pyrazines, cyanines, azulenes, azaazulenes and indocyanines, for example, are one of the few classes of small molecules having desirable photophysical properties for biomedical optical applications. These
compounds are very low molecular weight fluorescent scaffold systems with surprisingly bright emission in the yellow-to-red region of the electromagnetic spectrum. Binding of the dye component containing MMP inhibitors to MMPs may enhance fluorescence yield by immobilizing the dye component thereby minimizing non-radiative energy loss pathways.
[0164] In an embodiment of this aspect, the invention provides a method of using an optical agent, for example, in a biomedical procedure for optically imaging or visualizing a target tissue or a class of target tissues. The present methods include tissue selective imaging and visualization methods, such as imaging or visualization of a target tissue that expresses, secretes or otherwise produces MMPs, for example, a target tissue that expresses, secretes or otherwise produces the MMPs at an elevated level relative to production corresponding to a health (i.e. non-disease) condition. A method of this aspect comprises the step of administering a diagnostically effective amount of a compound to a subject, wherein the compound is a compound having any of formulas (FX1 ) - (FX113) or a pharmaceutical preparation thereof. In some embodiments, the method of this aspect further comprises contacting a target tissue that expresses, secretes or otherwise produces MMPs with a diagnostically effective amount of a compound having any of formulas (FX1 ) - (FX113). In some embodiments, the method of this aspect further comprises contacting cancer cells, for example, cancer cells of a tumor, with a diagnostically effective amount of a compound having any of formulas (FX1 ) - (FX113). In some embodiments, upon administration, the administered compound is optionally allowed to accumulate in a target region of interest (e.g., target cell, target tissue, target tumor, and/or target organ). The present methods are useful for imaging or visualizing colorectal cancer and other cancers, including prostate cancer, gastric cancer, esophageal cancer, uterine- endometrial cancer, pancreatic cancer, breast cancer, cervical cancer, head and neck cancer, hepatic cancer, skin cancer, gallbladder cancer, lung cancer and ovarian cancer.
[0165] In methods of this aspect, the compound that has been administered to the subject is exposed in vivo to electromagnetic radiation, thereby generating electromagnetic radiation emitted or scattered by the administered compound at the site of the target tissue. The electromagnetic radiation emitted or scattered by the administered compound is then detected either visually or using optical detection instrumentation as known in the art. In some embodiments, fluorescence is excited from the compound (e.g., due to absorption of electromagnetic radiation exposed to the administered compound), optionally via multiphoton excitation processes. In an embodiment particularly useful for imaging and/or visualization, the method of this aspect further comprises: (i) exposing a compound, such as a compound having any one of formula (FX1 ) - (FX113), administered to the subject to electromagnetic radiation capable of exciting emission from the compound; and (ii) detecting and/or measuring the intensity of emission from the compound. In some embodiments, the methods of the present invention use fluorescence excitation via exposure to light having wavelengths selected over the range of 300-1300 nm. In some embodiments, the wavelength of the electromagnetic radiation corresponds to a peak in the absorption spectrum of the optically functional MMP inhibitor, for example is within 20 nanometers of a peak in the absorption spectrum of the administered compound. In some biomedical procedures, the target site is exposed to electromagnetic radiation having sufficient fluence and/or power sufficient to excite the optically functional MMP inhibitor so as to generate measurable fluorescence from the administered compound. Exposure to electromagnetic radiation and activation of the administered compound may occur during or after administration of the optically functional MMP inhibitor and accumulation at the target tissue. For example, optical coherence tomography (OCT) is an optical imaging technique compatible with the present compounds that allows high resolution cross sectional imaging of tissue microstructure. OCT methods use wavelengths of about 1280 nm. Use of electromagnetic radiation having wavelengths selected over the range of 700 nanometers to 1300 nanometers may be useful for some in situ optical imaging methods of the present invention, including biomedical applications for imaging organs, tissue and/or tumors, anatomical visualization, optical guided surgery and endoscopic procedures. Compounds of the present methods may function as contrast agents, optical probes and/or tracer elements. The methods of the present invention include in vivo, in vitro and ex vivo imaging and visualization. The present invention provides methods for a range of clinical procedures, including optical imaging methods and/or visualization guided surgery and/or endoscopic diagnostic, monitoring and therapeutic procedures.
[0166] Appropriate power and intensity of the electromagnetic radiation depends on the size, depth, and the pathology of the target tissue (e.g. tumor or lesion), as is known to one skilled in the art. In an embodiment, the fluence of the electromagnetic radiation is preferably, but not always, kept below 200 mW/cm2 to minimize undesirable thermal effects. The intensity, power, and duration of the illumination, and the wavelength of the electromagnetic radiation may vary widely depending on the body location, the lesions site, the effect to be achieved, etc. In an embodiment, the power of the applied electromagnetic radiation is preferably selected over the range of 1 - 500 mW/cm2 and optionally selected over the range of 1 - 200 mW/cm2. In an embodiment, the duration of the exposure to applied electromagnetic radiation is selected over the range of 1 seconds to 60 minutes. In an embodiment, the electromagnetic radiation is applied internally, for example via endoluminal and endoscopic techniques.
[0167] Another aspect of the invention is a method of treating cancer, inflammation or inflammation-associated disorders, the method comprising administering to a subject in need thereof a therapeutically-effective amount of a compound having any of formulas (FX1 ) - (FX113) or a pharmaceutical preparation thereof. These disorders may include osteoarthritis and rheumatoid arthritis. In some embodiments, the method of this aspect further comprises contacting a target tissue that expresses, secretes or otherwise produces MMPs with a therapeutically effective amount of a compound having any of formulas (FX1 ) - (FX113). In some embodiments, the method of this aspect comprises contacting cancer cells, for example, cancer cells of a tumor, with a therapeutically effective amount of a compound having any of formulas (FX1 ) - (FX113). In some embodiments, upon administration, the administered compound is optionally allowed to accumulate in a target region of interest (e.g., target tissue, tumor, or organ). The invention provides compositions and methods to treat or inhibit the onset, development or recurrence of cancer. In an embodiment, for example, the invention provides compounds and methods for attenuating tumor progression and metastasis in animal tissues, comprising contacting a target tissue, such as tumor cells or tissues, with a therapeutically effective amount of an MMP inhibitor, such as a compound having any of formulas (FX1 ) - (FX113). In another aspect, the invention provides a method of reducing the risk of recurrence of a cancer or tumor after medical intervention (such intervention to include but not be limited to surgery, e.g. pulmonary surgery, surgical and endoscopic procedures, e.g. colonoscopy, gastrolaparoscopy, chemotherapy, etc.), comprising administering to a cancer patient an MMP inhibitor, such as a compound having any of formulas (FX1 ) - (FX113). Therefore, as will be apparent to one having skill in the art, the invention contemplates, for example, a method of minimizing post-operative recurrence of cancer in a patient, comprising administering to a cancer patient an MMP inhibitor, such as a compound having any of formulas (FX1 ) - (FX113). In an embodiment of the invention, the cancer is prostate cancer, gastrointestinal cancer, colon cancer, colorectal cancer, pancreatic cancer, breast cancer, lung cancer, throat cancer, skin cancer, stomach cancer, pancreatic cancer, brain cancer, liver cancer, prostate cancer, kidney cancer, bladder cancer, bone cancer, brain cancer, eye cancer, gallbladder cancer, head and neck cancer, Hodgkin lymphoma, mouth cancer, ovarian cancer, testicular cancer, throat cancer, esophageal cancer, small intestine cancer, pharyngeal cancer, laryngeal cancer, urethral cancer, uterine cancer, parathyroid cancer, cervical cancer, penile cancer or vaginal cancer. The compounds of the present invention may also be useful for the treatment of cancer in patients, as described above, either when used alone or in combination with one or more other anticancer agents, e.g., radiotherapy and/or other chemotherapeutic, including
antiangiogenic, treatments conventionally administered to patients for treating cancer. Several main categories and examples of such drugs are listed herein and include, but are not limited to, VEGF inhibitors, inhibitors of endothelial cell proliferation/migration, antagonists of angiogenic growth factors, inhibitors of Integrin/Survival signaling, and chelators of copper.
[0168] Methods of the invention may optionally further comprise a number of other steps. In an embodiment, the present methods further comprises the step of administering the optically functional MMP inhibitor into a bodily fluid of the subject. The optically functional MMP inhibitor may be introduced into the patient by any suitable method, including intravenous, intraperitoneal or subcutaneous injection or infusion, oral administration, transdermal absorption through the skin, or by inhalation. In an embodiment, the method further comprises contacting a target tissue, such as an organ, tissue, tumor, lesion, or cell type, with a compound of any one of formulas (FX1 ) - (FX113) prior to or during the exposure step. In an embodiment, the method further comprises allowing the compound to accumulate in a target tissue prior to exposure of the optically functional MMP inhibitor to electromagnetic radiation. In an embodiment, the method further comprises targeting the diagnostic agent to a selected organ, tissue, tumor, lesion, inflammation, or cell type. In an embodiment, the optically functional MMP inhibitor is administered to the skin, a tumor, surgical site, or a wound site. In an embodiment, for example, the optically functional MMP inhibitor is administered and/or delivered to a blood vessel, lung, heart, throat, ear, rectum, bladder, stomach, intestines, esophagus, liver, brain, prostrate, breast, or pancreas of the subject.
Example 4: Synthesis of Optically Functional MMP inhibitors
[0169] Certain optically functional MMP inhibitors of the invention can be synthesized according to the methods in references known to the art and using the provided procedures and modifications thereof known to one of ordinary skill in the art.
[0170] Synthesis of certain sulfone hydroxamic acids and hydroxamates is provided in references including: (1 ) Becker et al:. Synthesis and Structure - Activity Relationships of β- and a-Piperidine Sulfone Hydroxamic Acid Matrix Metalloproteinase Inhibitors with Oral
Antitumor Efficacy. J Med Chem, 2005, 48(21 ), 6713-6730; Becker et al..: a-Amino-3-sulfone hydroxamates as potent MMP-13 inhibitors that spare MMP-1 . Bioorg Med Chem Lett 2001 , 1 1 , 2719-2722; (2) Becker et al..: a-Alkyl-a-amino-3-sulfone hydroxamates as potent MMP inhibitors that spare MMP-1. Bioorg Med Chem Lett 2001 , 1 1 , 2723-2725, (3) Becker, et. al., J. Med Chem. 2005, 49, 6713-6730; (4) US Patent Publication No. 2001/0039287, published on Nov. 8, 2001 , corresponding to US Patent Application No. 09/256,948; (5) US Patent Publication No. 2001/0039287, published on Nov. 8, 2001 , corresponding to US Patent Application No. 09/256,948, filed on February 24, 1999; and (6) US Patent Publication No. 2002/0177588, published on Nov. 28, 2002, corresponding to US Patent Application No. 09/954,451 , filed on September 17, 2001. [0171] Synthesis of certain pyrazine derivatives is provided in references including: (1 ) Shirai, K. et al, Synthesis of fluorescent properties of 2,5-diamino-3,6-dicyanopyrazine dyes. Dyes and Pigments 1998, 39(1 ), 49-68; (2) Kim, J. H. et al., Self-assembling of aminopyrazine fluorescent dyes and their solid state spectra. Dyes and Pigments 1998 39(4), 341-357; and (3) Barlin, G. B., The pyrazines. In The Chemistry of Heterocyclic Compounds. A. Weissberger and E.C. Taylor, Eds. John Wiley & Sons, New York: 1982.
Example 4A: Preparation of an optically functional pyrazine-containing MMP inhibitor having formula (FX106)
[0172] An optically functional MMP inhibitor of the invention having formula (FX106) was prepared via the following procedure.
Step 1
Figure imgf000082_0001
[0173] A 250 mL RBF equipped with magnetic stir bar and dry condenser was charged with the 3g (7.2 mmole), of the fluoride, 1.9 g (10.8 mmole) phenol and 3.5 g (10.8 mmole) CSC03.
The reaction mixture was heated to 90^C for 6 hours then turned down to 60 ^C and let stir overnight. The next morning HPLC shows no starting material at 7.96 minutes and a new peak 8.88 minutes using a 5-95 gradient. The DMF was removed and the reaction was partionned between EA - 1 N aq. HCI. The organics were washed with 10 % IS^CC^, brine, dried, and concentrated in vacuo to 4.33 g. The crude product was purified by Flash chromatography: 25 % EA-H. Fractions 18-21 concentrated to 2.9 g clear oil that will turn to foam on vacuum drying. H-NMR (CDCI3) provided confirmation.
Step 2
Figure imgf000082_0002
[0174] A 250 ml RBF equipped with magnetic stir bar was charged with 2.9 g of the Boc compound 35 ml dichloroethane / and 35 ml trifluroacetic acid. After 1 hour the reaction was complete by LCMS analysis. The solvent were removed and the crude product was chased the 2X 50 ml of Heptane. The product was vacuum dried to afford 3.1 g of TFA salt. The product turns into a clear glass in the freezer.
Step 3 Symmetrical
products
Figure imgf000083_0001
Desired product
[0175] A 100 ml RBF was charged with 470 mg (1 eq.) diacid, 1.4 g amine TFA salt (1 eq.), 169 mg(1 eq.) pyyrolidinel g EDC (2.2 eq.), 705 mg (2.2 eq.) 528 ( 2.2 eq. ) triethylamine in 40 ml dry DMF. The reaction mixture was stirred at room temperature overnight. The next morning LCMS shows di-pyrrolidine at 3.5 minutes, desired product at 4.9 minutes, and di sulfonamide at 5.4 minutes. TLC (100 % EA) shows 3 yellow spots; di-sulfonamide moving at solvent front. The reaction was concentrated in vacuo and the residue was partionned between EA and satd. aq. bicarb (had to add brine to break up the emulsion). The organics were washed with aq. Citric, Brine, and dried. The EA solution was concentrated and purified by Flash Chromatography (3:1 EA:H) and it took 300 ml until first yellow spot came off. Fractions 1-5 clean di-sulfonamide (M + H = 1 109 g/mol), fractions 6-10 show a another yellow spot 5.1 minutes M + H = 951 g/mol (not sure what this is). Fraction 16 -26 were desired product (M + H = 707 g/mol). Fractions 1 -5 concentrated and vacuum dried to afford 210 mg yellow oil that solidified on pump. Fractions 16-26 concentrated and vacuum dried to afford 270 mg yellow oil that also turned into a glass / solid and was used as is.
Step 4
Figure imgf000083_0002
[0176] A 50 ml RBF equipped with magnetic stir bar was charged with 270 mg ester in 5 ml DMF. To this was added 37 mg ( 4.0 eq.) of lithium hydroxide in 1 .5 ml water. The reaction was stirred at room temperature. After 2 hours there was no reaction so the sample was heated to 60 ^C for 3 hours. Some reaction was observed and the sample was stirred stir at room temperature overnight. Next morning reaction was close to 1 :1 prod : st. mat. Sample was heated to 45 ^C and monitored throughout the day. After 4 hrs the starting material was essentially gone. The reaction mixture was partionned between EA and aq. Citric acid and the organics were washed with brine, dried and concentrated in vacuo to crude acid that was used as is.
Step 5.
Figure imgf000084_0001
[0177] A 50 ml RBF equipped with magnetic stir bar was charged with 285 mg crude acid, 74 mg (1.5 eq) OTHP-hydroxyl amine, 121 mg ( 1 .5 eq.) EDC, 85 mg (1.5 eq.) HOBt, 127 mg ( 3.0 eq.) TEA in 5 ml dry DMF. The reaction was heated to 40 °C and monitored by LCMS. After 2 hours the reaction only 60 % complete so a second 1 .5 eq. of hydroxyl amine, TEA and EDC were added. LCMS analysis after 2 hours showed essentially no starting material and product (two peaks ~ 8:1 of the two diastereomers). The reaction was partionned between EA and aq Citric acid. The organics were washed with satd. aq. bicarb, brine, dried, and then concentrated in vacuo to a yellow oil that was vacuum dried to 270 mg. Flash chromatography with EA - 5% MeOH / EA yielded fractions 1-3 (starting ethyl ester from page 128), fractions 4-6 contained product (2 diastereomers). The fractions were concentrated in vacuo and vacuum dried to afford 140 mg dry foam that was used directly in the next step.
Step 6.
Figure imgf000085_0001
[0178] A 50 ml RBF was charged with 140 mg OTHP-hydroxamate in 1 ml THF. To this solution was added 375 ml (2.6 eq.) of 1.25 M HCI-MEOH and the reaction was stirred at RT. LCMS analysis after 1 hour showed complete reaction. The reaction was concentrated in vacuo and vacuum dried to 125 mg's light red solid. HRMS (obs) M + H = 694.1920 g/mol.
[0179] HRMS (^ο^ΐ) M + H = 694.1901 g/mol.
Example 4B: Preparation of an optically functional pyrazine-containing MMP inhibitor having formula (FX107)
[0180] An optically functional MMP inhibitor of the invention having formula (FX107) was prepared via the following procedure.
Step 1
Symmetrical products
Figure imgf000085_0002
[0181] A 100 ml RBF equipped with magnetic stir bar was charged with 1.0 gram (5.05 mmole) diacid, 704 mg ( 5.05 mmole) beta-alanine methyl ester HCI salt, 379 mg (5.05 mmole) t-butyl amine, 1.53 g (15.15 mmole) Triethylamine, 4.3 g (1 1.2 mmole) HATU in 75 dry DMF (HATU last) and stirred at room temperature overnight. The reaction was concentrated in vacuo and partionned between EA and aq. Citric acid. The organics were washed with satd. aq.
bicarb, brine, dried and concentrated to crude. LCMS show majority of product is di-t-butyl amide some desired and very little di-beta Ala which may indicate the ester makes the amine much less reactive and when this is re-run the stiochemetry of the 2 amines may need adjusting (e.g., 2:1 stoichiometry). The crude was purified by flash chromatography (1 :1 EA:H) . The di t- butyl eluted first and started crystalizing as it came off the column; Heartcut frac 3-8.
Approximately 500 mg of this material was obtained. The desired product came out fractions 20-28 and it was concentrated to an orange solid that was vacuum dried to afford 210 mgs of (by LCMS) appears to be ~ 80-20 product to bis-t-butyl amide. This was purified via RPHPLC and the product containing fractions were concentrated to yield 150 mg's yellow solid. HRMS (theo) M + H = 339.1775 g/mol. HRMS (obs) M+H = 339.1780 g/mol. The material was used as is.
Step 2
Figure imgf000086_0001
[0182] A 100 ml RBF equipped with magnetic stir bar was charged with the 140 mg of ester, 74 mg (4 eq.) LiOH and stirred at room temperature. After 2 hours the reaction was complete. The reaction mixture was partionned between EA and 10 % KHSO4. The organics were washed with brine, dried and concentrated to the 82 mgs of acid that will be used directly in the next step.
Step 3
Figure imgf000086_0002
[0183] A 100 ml RBF equipped with magnetic stir bar was charged with 82 mg acid (1.0 eq.), 155 mg (1.2 eq.) of the amine HCI salt for step 2 of the previous prep of MP 3236, 58 mg (1 .2 eq.) EDC, 51 mg (1.5 eq.) HOBt, and 77 mg (3 eq.) triethylamine, in 8 ml DMF and stirred at room temperature overnight. LCMS shows complete consumption of acid, trace of amine (of course) and product. TLC (100 % EA) has product with Rf ~ 0.6 some slower moving trace impurities and amine at origin. Amine was removed by filtering through a plug of silica, which worked to remove amine at origin. This material was used as is.
Step 4
Figure imgf000086_0003
[0184] A 100 ml RBF equipped with magnetic stir bar was charged with 197 mg ester, 42 mg
(4.4 eq.) LiOH in 5 ml water and 15 mi's of THF. The reaction mixture was heated to 75^C and monitored by LCMS. After 8 hours there may be ~ 25 % conversion. 100 mg more base was added in 1 ml water and the reaction mixture left overnight. Next morning reaction ~ 4:1 ratio of product to starting material was observed with some trace impurities building in. 50 mgs more base was added. The reaction proceeded an additional hour then quenched and partionned between EA and 10 % KHSO4. The aqueous layer was yellow and it was washed with 75 ml
CHCI3. The combined organics were dried and dissolved into 6 m's DMF, filtered, and purified by RPHPLC. The product containing fractions were concentrated in vacuo and vacuum dried to afford 85 mg orange glass. LCMS M + H = 752 g/mol.
Step 5
Figure imgf000087_0001
[0185] A 100 ml RBF flask equipped with magnetic stir bar was charged with 82 mg's acid, 20 mg (1.5 eq.) protected hydroxyl amine, 34 mg (1.5 eq.) EDC, 23 mg (1.5 eq.) HOBt, 34 mg (3.0 eq.) triethylamine in 3 mi's dry DMF. The reaction was stirred at room temperature. LCMS after 3 hours showed starting material (4.8 min.), product (5.1 min.) and active ester (5.4 min.) in ~ 40:40:20 ratio. The reaction was stirred overnight at room temperature. LCMS shows about 10 % starting material with a trace of active ester and the rest product. 20 mg additional EDC was added and ~ 20 mg more amine and let stir. This worked but there was a trace of acid so the sample was concentrated in vacuo and partionned between EA and 10 % aq. KHSO4. The organics were washed with aq. Bicarb., brine, dried, and concentrated to crude that was immediately de-protected. Assume 100 % yield.
Step 6
Figure imgf000087_0002
[0186] A tared 4 dram vial was charged with the above in 4 ml 4N HCI-Dioxane. The reaction was stirred 20 minutes and LCMS showed complete conversion to product. The reaction was concentrated in vacuo and further vacuum dried to afford 100 mg red/orange foam. LCMS shows clean M + H = 767 g /mol. A sample was submitted for HRMS. HRMS (theo) M + H =
767.2429 g/mol. HRMS (obs) M + H = 767.2426 g/mol.
Example 4C: Preparation of an optically functional pyrazine-containing MMP inhibitor having formula (FX108)
[0187] An optically functional MMP inhibitor of the invention having formula (FX108) was prepared via the following procedure.
Step 1
Figure imgf000088_0001
[0188] A 50 mL RBF equipped with magnetic stir bar was charged with 1.0 g (1.0 eq.) Bromo amine, 1 .6 g (2.2 eq.) methyl Iodide, and 3.7 g (2.2 eq.) CsC03 in 25 ml DMF. The reaction was stirred at RT. The reaction mixture got dark and was let to stir overnight. LC/MS looks like previous runs. The reaction was partionned between EA and aq. Citric. The organics were washed with satd. bicarb, brine, dried and concentrated in vacuo to crude. Purification was done by Flash Chromatography (20 - 50 % EA-H). The fractions containing visible yellow product (even if some had some long wave impurities) were combined and concentrated then vacuum dried to afford 940 mgs of crystalline solid.
Step 2
Figure imgf000088_0002
[0189] A 100 ml RBF equipped with magnetic stir bar was charged with 239 mg (1.0 eq.) bromide, 548 mg (1.2 eq.) amine, 1 .0 g (3.0 eq.) CsC03, 33 mg (0.03 eq.)
Tris(dibenzilidineacetone) Palladium chloroform complex, and 59 mg (0.09 eq.) rac BINAP. The mixture was pumped and purged 3X with Argon. The reaction was charged with 20 ml toluene and the reaction was heated to 85^C overnight. LCMS shows mostly both starting materials. 10 ml toluene along with another 30 mg Pd catalyst was added. LCMS showed product and more Pd and BINAP was added. The reaction mixture was let go over the weekend. The reaction was concentrated in vacuo and partionned between EA / 10% aq. KHSO4. The organics were washed with aq. bicarb, brine, dried and concentrated to crude which was dissolved in MeOH and filtered for RPHPLC purification. Material purified via RPHPLC and product containing fractions were concentrated and vacuum dried to afford 390 mg (60 %) red oil. λ (abs) = 427 nM. LCMS clean M + H = 620 g/ mol. 1 H & 1 3C- NMR (CDCI3) look great.
Submitted for HRMS. HRMS (theo) M + H = 620.1785 g/mol. HRMS (obs) M + H = 620.1785 g
/mol.
Step 3
Figure imgf000089_0001
[0190] A 100 ml RBF equipped with magnetic stir bar was charged with 380 mg ethyl ester and 103 mg (4.0 eq.) LiOH in a mixture of 8 ml THF and 2 ml water. The reaction mixture was heated to 65^C. After 2 hours LCMS shows > 95 % starting material and no desired product. Potential decomposition of starting material was observed. The reaction was stirred at room temperature overnight. The next morning the reaction was heated to 65 °C after 5 hours ~ 30 % conversion. 4 more eq. of base was added and the sample heated to 80^C overnight. The reaction was monitored all day by LCMS. At the end of the day the reaction looked ~ 85 % conversion with a new impurity starting to grow in. The heat was turned off and the sample let stir overnight. LCMS shows ~ 85 % conversion to product 5.0 min 5-95 (M + H = 592 g/mol), some starting ethyl ester at 5.35 and a impurity @ 5.2 M + H = 654, M + Na = 677 all abs. @
430 nM.
Step 4
Figure imgf000089_0002
[0191] A 100 ml RBF equipped with a magnetic stir bar was charged with 363 mg (1.0 eq.) crude acid, 108 mg (1.5 eq.) amine, 176 mg (1.5 eq.) EDC, 125 mg (1.5 eq.) HOBt, and 75 mg ( 1.2 eq.) triethylamine in 5 mi's DMF. The reaction mixture was stirred at room temperature overnight. Next morning the LCMS shows no starting material. The reaction was partionned between EA and satd. bicarb. The organics were washed with 10 % KHSO4, brine, dried and concentrated in vacuo and vacuum dried. The crude was used as is in the next step.
Step 5
Figure imgf000089_0003
[0192] A 100 ml RBF was charged with the crude OTHP hydroxamate in 8 ml 4N HCI- Dioxane. LCMS after 10 minutes shows product at 4.8 (M + H = 607 g/mol) and the later moving impurities from ester hydrolysis. The reaction was concentrated in vacuo and purified via RPHPLC (5-95). The appropriate fractions were combined and concentrated and vacuum dried to afford 103 mg sticky oil-glass. HRMS (theo) + H = 607.1581 g/mol. HRMS (obs) M +
H = 607.1580 g/mol.
Example 4D: Preparation of an optically functional indocyanine-containing MMP inhibitor having formula (FX105)
[0193] An optically functional MMP inhibitor of the invention having formula (FX105) was prepared as follows.
[0194] Preparation of the starting compound (p-fluro-Boc ethyl ester) is described in Becker, et. al., J. Med Chem. 2005, 49, 6713-6730.
-all reaction scheme is provided below.
Figure imgf000090_0001
[0196] A 100 ml RBF equipped with magnetic stir bar was charged with 573 mg (1 mmol) starting compound and 630 mg (15 mmol) lithium hydroxide monohydrate in 8 ml of THF and 2 ml of water. The reaction mixture was heated to reflux for 24 hours. The next day the reaction was found to be ~ 70 % converted to the desired acid. The product was concentrated to dryness and then partitioned between ethyl acetate and 10 % aqueous potassium hydrogen sulfate. The organics were washed with brine, dried and concentrated to crude that was purified via RPHPLC (5-95 acetonitrile-water with 0.1 % TFA in both mobile phases). The product containing fractions was concentrated and dried to afford 275 mg of product as a white solid that was used without further purification.
Step 2
Figure imgf000090_0002
[0197] A 100 ml RBF equipped with magnetic stir bar was charged with 255 mg (1.0 eq.) acid, 68 mg (1.25 eq.) amine, 122 mg (1.25 eq.) EDC, 95 mg (1.25 eq.) HOBt, and 95 mg (2.0 eq.) TEA in 10 ml of dry DMF. The reaction mixture was stirred at room temperature overnight. The next morning LCMS analysis showed > 90 % conversion to product. The reaction was partitioned between EA and saturated aqueous sodium bicarbonate. The organics were washed with 10 % aq. Citric acid, saturated aqueous sodium bicarbonate, brine, and dried. The organics were concentrated to crude solid, ~ 300 mg, and used without further purification. HRMS(theo) M + H = 645.2088 g/mol . HRMS(obs) M + H = 645.2072 g/mol.
Step 3
Figure imgf000091_0001
[0198] A 100 ml RBF equipped with magnetic stir bar was charged with 300 mg crude starting material with 6 ml 4 N-HCI / Dioxane and stirred one hour until LCMS showed complete reaction. The cloudy mixture was concentrated in vacuo to produce an off white solid that was vacuum dried to afford - 180 mg off white solid. HRMS(theo) M + H = 461.0989 g/mol.
HRMS(obs) = M + H = 461 .0989 g/mol.
Step 4
Figure imgf000091_0002
[0199] A 100 ml RBF equipped with magnetic stir bar was charged with 276 mg (1.0 eq.) of the chloride, 171 mg (1.0 eq.) of the amine hydrochloride and 70 mg (2.0) eq.) triethylamine in 10 ml of dry DMF. The green solution was heated to 400°C. After 40 minutes LCMS showed starting material (4.4 minute retention time) and 3 new peaks. One was at 4.1 and is blue shifted to 713 nM and had a smaller MW than starting material. There was a small peak at 4.65 that gave the correct M + H, and a peak at 4.8 with M + H = 1009 g/mol. The temperature was increased to 600°C. After 30 minutes starting material was gone and the earlier peak M + H = 709 g/mol had grown. This indicated replacement of CI by OH. There was a trace of desired product and a good amount of a later impurity with a M + H = 1009 g/mol. This was concentrated to 1/2 volume and was put on RPHPLC. The RPHPLC separated the 3 peaks. The desired product (~ 10 mg deep blue solid) was submitted for HRMS. HRMS(theo) M + H = 1 151 .3786 g/mol. HRMS(obs) M + H = 1 151 .3790 g/mol. {abs) = > 700 nM, hMMP-2 = 2.76 nM, hMMP-9 = 1 .37 nM. Figure 6A provides a high resolution mass spectrum of the compound having formula (FX105). Figure 6B provides an expanded view of the mass spectrum (top panel) of the compound having formula (FX105) and a simulated mass spectrum (lower panel) for a mass to charge ratio (m/z) range of about 1 149 to about 1 154. Figure 6C provides an expanded view of the mass spectrum (top panel) of the compound having formula (FX105) and a simulated mass spectrum (lower panel) for a mass to charge ratio (m/z) range of about 574 to 576. As shown in Figures 6B an 6C, the observed mass spectrum agrees well with the simulated mass spectrum. [0200] The IC50 against MMP-2 for compound (FX105) was measured to be 2.76 nM. The IC50 against MMP-9 for compound (FX105) was measured to be 1 .37 nM.
Example 4E: Optically functional azulene-containing MMP inhibitor having formulas (FX110) and (FX111)
[0201] The invention provides optically functional azulene-containing MMP inhibitors being of the formula (FX110):
Figure imgf000092_0001
(FX110); wherein Y is -CH- or -N-, and R is hydrogen, -
OCF3, C C6 alkyl, C3-C6 cycloalkyl, C5-C10 aryl, C5-C10 heteroaryl, C C6 acyl, C2-C6 alkenyl, C2- C6 alkynyl, C5-C10 alkylaryl, halo, halomethyl, dihalomethyl, trihalomethyl, -CN, -C02 6°, - CONR61R62, -COR63, -NO2, -SOR64, -OSR65 , -S02R66, -S02OR67, -S02NR68R69, -PO3R70R71 ; -OR72, -SR73, -NR74R75, -NR76COR77, -CH2(CHOH)cR78,-(CH2CH20)cR79, c, PS1 or PS2, wherein R60 - R79, PS1 and PS2 are as set forth in the description of formulas (FX1 ) and (FX2). This compound is expected to have an absorption peak in the visible region of the
electromagnetic spectrum around 548 nm. In an embodiment, for example, the invention provides compounds having formula (FX110), wherein R is-OCF3. In an embodiment, invention provides optically functional azulene-containing MMP inhibitors being of the formula (FX111 ):
Figure imgf000092_0002
(FX111 ); wherein Y is -CH- or -N-, and wherein R is hydrogen, -OCF3, C C6 alkyl, C3-C6 cycloalkyl, C5-C10 aryl, C5-C10 heteroaryl, C C6 acyl, C2- C6 alkenyl, C2-C6 alkynyl, C5-C10 alkylaryl, halo, halomethyl, dihalomethyl, trihalomethyl, -CN, - C02R60, -CONR61 R62, -COR63, -N02, -SOR64, -OSR65 , -S02R66, -S02OR67, -S02NR68R69, - PO3R70R71, -OR72, -SR73, -NR74R75, -NR76COR77, -CH2(CHOH)cR78,-(CH2CH20)cR79, PS1 or PS2; wherein each of Q1 - Q4 is independently hydrogen or electron withdrawing groups and each of Z1 - Z3 is independently hydrogen or a halo group (e.g., fluoro, chloro, bromo, iodo, etc.); wherein R60 - R79, PS1 and PS2 are as set forth in the description of formulas (FX1 ) and (FX2). In an embodiment, for example, the invention provides compounds having formula
(FX111 ), wherein each of Z1 - Z3 is independently hydrogen or fluorine. In an embodiment, for example, the invention provides compounds having formula (FX111 ), wherein each of Q1 - Q4 is independently hydrogen or -CN, -C02R40, -S02OR43, -CONR52R53, -COR54, -N02, -SOR41, - S02R55, -POsR^R45, halo, C C6 acyl, trihalomethyl, or -S02NR56R57. In an embodiment, for example, the invention provides compounds having formula (FX111 ), wherein each of Q1 - Q4 is hydrogen, and wherein each of Z1 - Z3 is independently hydrogen or fluorine. In an
embodiment, for example, the invention provides compounds having formula (FX110), wherein R is-OCF3.
[0202] This class of azulene-containing MMP compounds may be prepared, for example, via peptide coupling to azulene acid. (See, e.g., Leermakers and Bowman. Structure vs reactivity in azulene carboxylic acids and esters. Journal of Organic Chemistry 1964, 29, 3708-371 1 .)
[0203] Example 4F: Optically functional MMP inhibitors having an optical dye in the S1 ' binding pocket.
[0204] The invention provides optically function MMP inhibitors having an optical dye in the S1 ' binding pocket having the formula (FX112) or (FX113):
Figure imgf000093_0001
R and R' is independently
H or C1-C10 alkyl, and optionally wherein each R and R' is independently H or CrC6 alkyl, and optionally wherein each R and R' is independently H or C1-C3 alkyl.
Example 5: Compositions and methods for phototherapy
[0205] The invention includes phototherapy methods wherein a phototherapeutic agent comprising a compound of any one of the formulas (FX1 ) - (FX113) is administered to a patient, for example, wherein a therapeutically effective amount of such a component is administered to a patient in need of treatment. In this aspect, compounds of the invention provide an optical agent capable of selectively targeting and delivery to tissue expressing MMPs and further functions as a phototherapeutic agent and MMP inhibitor. Upon administration, the phototherapeutic agent is allowed to accumulate in a target region of interest (e.g., target tissue, tumor, or organ). To induce selective tissue damage, the phototherapeutic agent is activated by exposure to electromagnetic radiation. In an embodiment, the phototherapeutic agent is activated after an effective concentration of the phototherapeutic agent has accumulated in a target tissue. An effective concentration of a compound of the invention depends on the nature of the formulation, method of delivery, target tissue, activation method and toxicity to the surrounding normal non-target tissue. Exposure to electromagnetic radiation and activation of the phototherapeutic agent may occur during or after administration of the phototherapeutic agent and accumulation at the target tissue.
[0206] For photoactivation, the target region is illuminated with electromagnetic radiation having a wavelength in the range of about 350 nm to about 1300 nm, preferably for some applications in the range of about 350 nm to about 900 nm. In some embodiments, the wavelength of the electromagnetic radiation corresponds to a peak in the absorption spectrum of the phototherapeutic agent, for example is within 20 nanometers of a peak in the absorption spectrum of the phototherapeutic agent in the visible or NIR regions. In some phototherapeutic procedures the target site is exposed to electromagnetic radiation having sufficient fluence and/or power sufficient to activate the phototherapeutic agent so as to induce cell death, for example via necrosis or apoptosis processes. In some embodiments, electromagnetic radiation of low energy, power or fluence is needed to activate the phototherapeutic agent. If the region of interest is, for example a lesion or tumor on the skin surface, the region can be directly illuminated. Otherwise, endoscopic and/or endoluminal catheters equipped with an
electromagnetic radiation source may be employed to provide a photodiagnostic and/or the phototherapeutic effect.
[0207] Appropriate power and intensity of the electromagnetic radiation depends on the size, depth, and the pathology of the lesion, as is known to one skilled in the art. In an embodiment, the fluence of the electromagentic radiation is preferably, but not always, kept below 200 mW/cm2 to minimize undesirable thermal effects. The intensity, power, and duration of the illumination, and the wavelength of the electromagnetic radiation may vary widely depending on the body location, the lesions site, the effect to be achieved, etc. In an embodiment, the power of the applied electromagnetic radiation is preferably selected over the range of 1 - 500 mW/cm2 and optionally selected over the range of 1 - 200 mW/cm2. In an embodiment, the duration of the exposure to applied electromagentic radiation selected over the range of 1 seconds to 60 minutes.
[0208] In an embodiment, the invention provides a method of using a phototherapeutic agent, the method comprising: (i) administering a therapeutically effective amount of a phototherapeutic agent to a subject, the phototherapeutic agent comprising a compound being of any one of formula (FX1 ) - (FX113), wherein at least one of X1 to X17, R1 to R5 and R8 to R10 is PS1 or PS2, or a pharmaceutically acceptable salt or ester thereof; and (ii) exposing the phototherapeutic agent administered to the patient to electromagnetic radiation. In an embodiment, for example, erapy being of formula (FX1 ) or (FX2):
(FX1 ); or
Figure imgf000095_0001
(FX2); or a pharmaceutically acceptable salt or ester thereof; wherein W1 - W3, A, L1 - L3, x, q, r, U, R1 to R5, and ring Z are defined as described above in connection with formulas (FX1 ) and (FX2), wherein at least one of R1 - R5 or A includes a group corresponding to PS1 or PS2, and wherein each PS1 is independently an azide, azo, diazo, oxaza, diaza, dithia, thioxa, or dioxa group; and each PS2 is independently a group corresponding to a porphyrin, benzoporphyrin, phthalocyanine, phenothiazine, chlorin, bacteriochlorin, phthalocyanine, porphyrin, purpurin, merocyanine, pheophorbides, psoralen, aminolevulinic acid (ALA), hematoporphyrin derivative, porphycenes, or porphacyanine. In an embodiment, for example, the invention provides compounds having formula (FX1 ) or (FX2) wherein at least one of R1 - R5 or A includes a Type 2 photosensitizer. Alternatively, the invention provides compounds having formula (FX1 ) or (FX2) wherein at least one of R1 - R5 or A includes a Type 1 photosensitizer. In an embodiment, of this aspect, the invention provides a compound having formula (FX1 ) or (FX2) wherein R1 and R5 are each hydrogen. In an embodiment, the method of the invention comprises administering to a patient a compound having any one of formula selected from (FX1 ) - (FX113), including any of the specific compositions classes and compounds described in connection with formula (FX1 ) - (FX113), particularly those compounds having a photosensitizer component.
[0209] In an embodiment, the phototherapeutic agent is exposed to a therapeutically effective amount of electromagnetic radiation. As used herein, a therapeutically effective amount of electromagentic radiation is an amount for achieving a desired therapeutic result, for example an amount for generating a therapeutically effective amount of reactive species for damaging or causing cell death of a selected target tissue. In an embodiment, the method further comprises generating one or more reactive species from said compound administered to the patient via the exposure of the phototherapeutic agent to applied electromagnetic radiation. In an embodiment, for example, the method further comprises the step of cleaving one or more photolabile bonds of the optical agent so as to generate reactive species comprising free radicals. In an
embodiment, the method further comprises targeting the phototherapeutic agent to a selected organ in the patient or to a selected tissue type in the patient. In an embodiment, a
therapeutically effective dose of the phototherapeutic agent is administered to a patient in need of treatment.
[0210] Embodiments of this aspect, may comprise a method of carrying out an in vivo therapeutic and/or diagnostic procedure. In an embodiment, the invention comprises a method of carrying out an in vivo phototherapeutic, photoactivation, and/or photosensitizing procedure. The present methods have broad clinical utility which includes, but is not limited to, phototherapy of tumors, inflammatory processes, and impaired vasculature. In embodiments, subjects of the invention may be any mammal, such as a human, and optionally the subject of the present methods is a patient in need of treatment and/or diagnosis. The present methods are also useful in ex vivo and in vitro procedures, including medical therapeutic and diagnostic procedures.
[0211] Methods of the invention may optionally further comprise a number of other steps. In an embodiment, the method further comprises the step of administering the phototherapeutic agent into a bodily fluid of the subject. The phototherapeutic agent may be introduced into the patient by any suitable method, including intravenous, intraperitoneal or subcutaneous injection or infusion, oral administration, transdermal absorption through the skin, or by inhalation. In an embodiment, the method further comprises contacting a target tissue, such as an organ, tissue, tumor, lesion, or cell type, with a compound of any one of formulas (FX1 ) - (FX113) prior to or during the exposure step. In an embodiment, the method further comprises allowing the compound to accumulate in a target tissue prior to exposure of the phototherapeutic agent to electromagnetic radiation. In an embodiment, the method further comprises targeting the diagnostic agent to a selected organ, tissue, tumor, lesion, inflammation, or cell type. In an embodiment, the phototherapeutic agent is administered to the skin, a tumor, surgical site, or a wound site. In an embodiment, for example, the phototherapeutic agent is administered and/or delivered to a blood vessel, lung, heart, throat, ear, rectum, bladder, stomach, intestines, esophagus, liver, brain, prostrate, breast, or pancreas of the subject.
[0212] As will be understood by one having skill in the art, the optical conditions for the step of exposing the phototherapeutic agent administered to the patient to electromagnetic radiation will vary considerably with the (i) therapeutic and/or diagnostic objectives, and (ii) the condition of the subject (e.g., height, weight, state of health etc.). In an embodiment, the applied
electromagnetic radiation has wavelengths, energy and/or fluence sufficient to achieve a desired therapeutic and/or diagnostic result. In an embodiment, the electromagnetic radiation has wavelengths, energy and/or fluence sufficient to activate the phototherapeutic agent, for example wavelengths, energy and/or fluence sufficient to result in generation of reactive species, including singlet oxygen, ions, and/or free radicals. In an embodiment, the electromagnetic radiation has wavelengths, energy and/or fluence sufficient to result in cleavage of at least one photolabile bond of the optical agent upon absorption. In an embodiment, the electromagnetic radiation exposed to the phototherapeutic agent has wavelengths
corresponding to a maximum in the absorption spectrum of the phototherapeutic agent, preferably for some applications a maximum in the visible or NIR regions of the electromagnetic spectrum. Optionally, excitation is achieved using electromagnetic substantially free (e.g., less than about 10% of total radiant energy), of ultraviolet radiation, for example, to minimize exposure of the subject to electromagnetic radiation capable of causing unwanted cell or tissue damage. Electromagnetic radiation may be provided to the phototherapeutic agent using a range of optical sources and/or surgical instrumentation, including a laser, light emitting diodes, fiber optic device, endoscope, catheter, optical filters, or any combination of these.
Example 4: ADMINISTRATION AND FORMULATION
4a: Salts and Prodrugs
[0213] The invention contemplates pharmaceutically active compounds either chemically synthesized or formed by in vivo biotransformation to compounds set forth herein.
[0214] Compounds of this invention and compounds useful in the methods of this invention include those of the compounds and formula(s) described herein and pharmaceutically- acceptable salts and esters of those compounds. In embodiments, salts include any salts derived from the acids and bases of the formulas herein which are acceptable for use in human or veterinary applications. In embodiments, the term ester refers to hydrolyzable esters of compounds of the names and formulas herein. In embodiments, salts and esters of the compounds of the formulas herein can include those which have the same or better therapeutic, diagnostic, or pharmaceutical (human or veterinary) general properties as the compounds of the formulas herein. In an embodiment, a composition of the invention is a compound or salt or ester thereof suitable for pharmaceutical formulations.
[0215] Compounds of the invention can have prodrug forms. Prodrugs of the compounds of the invention are useful in embodiments including compositions and methods. Any compound that will be converted in vivo to provide a biologically, pharmaceutically, diagnostically, or therapeutically active form of a compound of the invention is a prodrug. Various examples and forms of prodrugs are well known in the art. Examples of prodrugs are found, inter alia, in: Design of Prodrugs, edited by H. Bundgaard, (Elsevier, 1985); Methods in Enzymology, Vol. 42, at pp. 309-396, edited by K. Widder, et. al. (Academic Press, 1985); A Textbook of Drug Design and Development, edited by Krosgaard-Larsen and H. Bundgaard, Chapter 5, "Design and Application of Prodrugs," by H. Bundgaard, at pp. 1 13-191 (1991 ); H. Bundgaard, Advanced Drug Delivery Reviews, Vol. 8, p. 1 -38 (1992); H. Bundgaard, et al., Journal of Pharmaceutical Sciences, Vol. 77, p. 285 (1988); and Nogrady (1985) Medicinal Chemistry A Biochemical Approach, Oxford University Press, New York, pages 388-392). A prodrug, such as a pharmaceutically acceptable prodrug, can represent prodrugs of the compounds of the invention which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic response, and the like, commensurate with a reasonable benefit/risk ratio, and effective for their intended use. Prodrugs of the invention can be rapidly transformed in vivo to a parent compound of a compound described herein, for example, by hydrolysis in blood or by other cell, tissue, organ, or system processes. Further discussion is provided in: T. Higuchi and V. Stella, Pro-drugs as Novel Delivery Systems, V. 14 of the A.C.S. Symposium Series; and in Edward B. Roche, ed., Bioreversible Carriers in Drug Design, American Pharmaceutical Association and Pergamon Press (1987).
[0216] Optical agents of the invention can be formulated with pharmaceutically-acceptable anions and/or cations. Pharmaceutically-acceptable cations include among others, alkali metal cations (e.g., Li+, Na+, K+), alkaline earth metal cations (e.g., Ca2+, Mg2+), non-toxic heavy metal cations and ammonium (NH4 +) and substituted ammonium (N(R')4 +, where R' is hydrogen, alkyl, or substituted alkyl, i.e., including, methyl, ethyl, or hydroxyethyl, specifically, trimethyl ammonium, triethyl ammonium, and triethanol ammonium cations). Pharmaceutically-acceptable anions include, among others, halides (e.g., F", CI", Br", At"), sulfate, acetates (e.g., acetate, trifluoroacetate), ascorbates, aspartates, benzoates, citrates, and lactate.
[0217] Pharmaceutically acceptable salts comprise pharmaceutically-acceptable anions and/or cations. As used herein, the term "pharmaceutically acceptable salt" can refer to acid addition salts or base addition salts of the compounds in the present disclosure. A pharmaceutically acceptable salt is any salt which retains at least a portion of the activity of the parent compound and does not impart significant deleterious or undesirable effect on a subject to whom it is administered and in the context in which it is administered. Pharmaceutically acceptable salts include metal complexes and salts of both inorganic and organic acids. Pharmaceutically acceptable salts include metal salts such as aluminum, calcium, iron, magnesium, manganese and complex salts. Pharmaceutically acceptable salts include, but are not limited to, acid salts such as acetic, aspartic, alkylsulfonic, arylsulfonic, axetil, benzenesulfonic, benzoic, bicarbonic, bisulfuric, bitartaric, butyric, calcium edetate, camsylic, carbonic, chlorobenzoic, cilexetil, citric, edetic, edisylic, estolic, esyl, esylic, formic, fumaric, gluceptic, gluconic, glutamic, glycolic, glycolylarsanilic, hexamic, hexylresorcjnoic, hydrabamic, hydrobromic, hydrochloric, hydroiodic, hydroxynaphthoic, isethionic, lactic, lactobionic, maleic, malic, malonic, mandelic, methanesulfonic, methylnitric, methylsulfuric, mucic, muconic, napsylic, nitric, oxalic, p- nitromethanesulfonic, pamoic, pantothenic, phosphoric, monohydrogen phosphoric, dihydrogen phosphoric, phthalic, polygalactouronic, propionic, salicylic, stearic, succinic, sulfamic, sulfanlic, sulfonic, sulfuric, tannic, tartaric, teoclic, toluenesulfonic, and the like. Pharmaceutically acceptable salts can be derived from amino acids, including, but not limited to, cysteine. Other pharmaceutically acceptable salts can be found, for example, in Stahl et al., Handbook of Pharmaceutical Salts: Properties, Selection, and Use, Wiley-VCH, Verlag Helvetica Chimica Acta, Zurich, 2002. (ISBN 3-906390-26-8).
4b: Efficacy
[0218] Typically, a compound of the invention, or pharmaceutically acceptable salt thereof, is administered to a subject in a diagnostically or therapeutically effective amount. One skilled in the art generally can determine an appropriate dosage.
[0219] Compositions for oral administration can be, for example, prepared in a manner such that a single dose in one or more oral preparations contains at least about 20 mg of the present compound per square meter of subject body surface area, or at least about 50, 100, 150, 200, 300, 400, or 500 mg of the present compound per square meter of subject body surface area (the average body surface area for a human is, for example, 1.8 square meters). In particular, a single dose of a composition for oral administration can contain from about 20 to about 600 mg, and in certain aspects from about 20 to about 400 mg, in another aspect from about 20 to about 300 mg, and in yet another aspect from about 20 to about 200 mg of the present compound per square meter of subject body surface area. Compositions for parenteral administration can be prepared in a manner such that a single dose contains at least about 20 mg of the present compound per square meter of subject body surface area, or at least about 40, 50, 100, 150, 200, 300, 400, or 500 mg of the present compound per square meter of subject body surface area. In particular, a single dose in one or more parenteral preparations contains from about 20 to about 500 mg, and in certain aspects from about 20 to about 400 mg, and in another aspect from about 20 to about 450 mg, and in yet another aspect from about 20 to about 350 mg of the present compound per square meter of subject body surface area. It should be recognized that these oral and parenteral dosage ranges represent generally preferred dosage ranges, and are not intended to limit the invention. The dosage regimen actually employed can vary widely, and, therefore, can deviate from the generally preferred dosage regimen. It is contemplated that one skilled in the art will tailor these ranges to the individual subject.
[0220] Toxicity and therapeutic efficacy of such compounds and bioconjugates can be determined by standard pharmaceutical procedures in cell cultures or experimental animals for determining the LD50 (the dose lethal to 50% of the population) and the ED50, (the dose therapeutically effective in 50% of the population). The dose ratio between toxic and therapeutic effects is the therapeutic index that can be expressed as the ratio LD5o/ED5o. Compounds and bioconjugates that exhibit large therapeutic indices are preferred. While compounds and bioconjugates exhibiting toxic side effects can be used, care should be taken to design a delivery system that targets such compounds and bioconjugates to the site affected by the disease or disorder in order to minimize potential damage to unaffected cells and reduce side effects. [0221] Data obtained from the cell culture assays and animal studies can be used in formulating a range of dosages for use in humans and other mammals. The dosage of such compounds and bioconjugates lies preferably within a range of circulating plasma or other bodily fluid concentrations that include the ED50 and provides clinically efficacious results (i.e., reduction in disease symptoms). The dosage can vary within this range depending upon the dosage form employed and the route of administration utilized. For any compound and bioconjugate of the present invention, the therapeutically effective amount can be estimated initially from cell culture assays. A dosage can be formulated in animal models to achieve a circulating plasma concentration range that includes the ED50 (the concentration of the test compound that achieves a half-maximal inhibition of symptoms) as determined in cell culture. Such information can be used to more accurately determine useful dosages in humans and other mammals. Compound and bioconjugate levels in plasma can be measured, for example, by high performance liquid chromatography.
[0222] An amount of a compound or bioconjugate that can be combined with a pharmaceutically acceptable carrier to produce a single dosage form will vary depending upon the patient treated and the particular mode of administration. It will be appreciated by those skilled in the art that the unit content of a compound/bioconjugate contained in an individual dose of each dosage form need not in itself constitute a therapeutically effective amount, as the necessary therapeutically effective amount could be reached by administration of a number of individual doses. The selection of dosage depends upon the dosage form utilized, the condition being treated, and the particular purpose to be achieved according to the determination of those skilled in the art.
[0223] The dosage and dosage regime for treating a disease or condition can be selected in accordance with a variety of factors, including the type, age, weight, sex, diet and/or medical condition of the patient, the route of administration, pharmacological considerations such as activity, efficacy, pharmacokinetic and/or toxicology profiles of the particular compound/bioconjugate employed, whether a compound/bioconjugate delivery system is utilized, and/or whether the compound/bioconjugate is administered as a pro-drug or part of a drug combination. Thus, the dosage regime actually employed can vary widely from subject to subject, or disease to disease and different routes of administration can be employed in different clinical settings.
[0224] The identified compounds/bioconjugates monitor, treat, inhibit, control and/or prevent, or at least partially arrest or partially prevent, diseases and conditions of interest and can be administered to a subject at therapeutically effective amounts and optionally diagnostically effective amounts. Compositions/formulations of the present invention comprise a therapeutically effective amount (which can optionally include a diagnostically effective amount) of at least one compound or bioconjugate of the present invention. Subjects receiving treatment that includes a compound/bioconjugate of the invention are preferably animals (e.g., mammals, reptiles and/or avians), more preferably humans, horses, cows, dogs, cats, sheep, pigs, and/or chickens, and most preferably humans.
4c: Administration
[0225] The preferred composition depends on the route of administration. Any route of administration can be used as long as the target of the compound or pharmaceutically acceptable salt is available via that route. Suitable routes of administration include, for example, oral, intravenous, parenteral, inhalation, rectal, nasal, topical (e.g., transdermal and intraocular), intravesical, intrathecal, enteral, pulmonary, intralymphatic, intracavital, vaginal, transurethral, intradermal, aural, intramammary, buccal, orthotopic, intratracheal, intralesional, percutaneous, endoscopical, transmucosal, sublingual, and intestinal administration.
[0226] In an embodiment, the invention provides a method for treating a medical condition comprising administering to a subject (e.g. patient) in need thereof, a therapeutically effective amount of a composition of the invention, such as a compound of any one of formulas (FX1 ) - (FX113). In an embodiment, the invention provides a method for diagnosing or aiding in the diagnosis of a medical condition comprising administering to a subject in need thereof, a diagnostically effective amount of a composition of the invention. In an embodiment, the medical condition is cancer, or various other diseases, injuries, and disorders, including cardiovascular disorders such as atherosclerosis and vascular restenosis, inflammatory diseases, ophthalmic diseases and dermatological diseases.
[0227] The diagnostic and therapeutic formulations of this invention can be administered alone, but can be administered with a pharmaceutical carrier selected upon the basis of the chosen route of administration and standard pharmaceutical practice.
[0228] Any suitable form of administration can be employed in connection with the diagnostic and therapeutic formulations of the invention. The diagnostic and therapeutic formulations of this invention can be administered intravenously, in oral dosage forms, intraperitoneally, subcutaneously, or intramuscularly, all using dosage forms well known to those of ordinary skill in the pharmaceutical arts.
[0229] The present compositions, preparations and formulations can be formulated into diagnostic or therapeutic compositions for enteral, parenteral, topical, aerosol, inhalation, or cutaneous administration. Topical or cutaneous delivery of the compositions, preparations and formulations can also include aerosol formulation, creams, gels, solutions, etc. The present compositions, preparations and formulations are administered in doses effective to achieve the desired diagnostic and/or therapeutic effect. Such doses can vary widely depending upon the particular compositions employed in the composition, the organs or tissues to be examined, the equipment employed in the clinical procedure, the efficacy of the treatment achieved, and the like. These compositions, preparations and formulations contain an effective amount of the composition(s), along with conventional pharmaceutical carriers and excipients appropriate for the type of administration contemplated. These compositions, preparations and formulations can also optionally include stabilizing agents and skin penetration enhancing agents.
(i) Parenteral Administration
[0230] Compounds and bioconjugates of the present invention can be formulated for parenteral administration by injection (e.g., by bolus injection or continuous infusion). Formulations for injection can be presented in unit dosage form in ampoules or in multi-dose containers with an optional preservative added. The parenteral preparation can be enclosed in ampoules, disposable syringes or multiple dose vials made of glass, plastic or the like. The formulation can take such forms as suspensions, solutions or emulsions in oily or aqueous vehicles, and can contain formulatory agents such as suspending, stabilizing and/or dispersing agents.
[0231] For example, a parenteral preparation can be a sterile injectable solution or suspension in a nontoxic parenterally acceptable diluent or solvent (e.g., as a solution in 1 ,3-butanediol). Among the acceptable vehicles and solvents that can be employed are water, Ringer's solution, and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose any bland fixed oil can be employed including synthetic mono- or di-glycerides. In addition, fatty acids such as oleic acid can be used in the parenteral preparation.
[0232] Alternatively, compounds and bioconjugates of the present invention can be formulated in powder form for constitution with a suitable vehicle, such as sterile pyrogen-free water, before use. For example, a compound/bioconjugate suitable for parenteral administration can include a sterile isotonic saline solution containing between 0.1 percent and 90 percent weight per volume of the compound/bioconjugate. By way of example, a solution can contain from about 5 percent to about 20 percent, more preferably from about 5 percent to about 17 percent, more preferably from about 8 to about 14 percent, and still more preferably about 10 percent weight per volume of the compound/bioconjugate. The solution or powder preparation can also include a solubilizing agent and a local anesthetic such as lignocaine to ease pain at the site of the injection. Other methods of parenteral delivery of compounds/bioconjugates will be known to the skilled artisan and are within the scope of the invention.
(ii) Oral Administration
[0233] For oral administration, a compound/bioconjugate of the invention can be formulated to take the form of tablets or capsules prepared by conventional means with one or more pharmaceutically acceptable carriers (e.g., excipients such as binding agents, fillers, lubricants and disintegrants).
(iii) Controlled-Release Administration
[0234] Controlled-release (or sustained-release) preparations can be formulated to extend the activity of a compound/bioconjugate and reduce dosage frequency. Controlled-release preparations can also be used to effect the time of onset of action or other characteristics, such as blood levels of the compound/bioconjugate, and consequently affect the occurrence of side effects.
[0235] Controlled-release preparations can be designed to initially release an amount of a compound/bioconjugate that produces the desired therapeutic effect, and gradually and continually release other amounts of the compound/bioconjugate to maintain the level of therapeutic effect over an extended period of time. In order to maintain a near-constant level of a compound/bioconjugate in the body, the compound/bioconjugate can be released from the dosage form at a rate that will replace the amount of compound/bioconjugate being metabolized and/or excreted from the body. The controlled-release of a compound/bioconjugate can be stimulated by various inducers, e.g., change in pH, change in temperature, enzymes, water, and/or other physiological conditions or molecules.
[0236] Controlled-release systems can include, for example, an infusion pump which can be used to administer the compound/bioconjugate in a manner similar to that used for delivering insulin or chemotherapy to the body generally, or to specific organs or tumors. Typically, using such a system, the compound/bioconjugate is administered in combination with a biodegradable, biocompatible polymeric implant that releases the compound/bioconjugate over a controlled period of time at a selected site. Examples of polymeric materials include polyanhydrides, polyorthoesters, polyglycolic acid, polylactic acid, polyethylene vinyl acetate, and copolymers and combinations thereof. In addition, a controlled release system can be placed in proximity of a therapeutic target (e.g., organ, tissue, or group of cells), thus requiring only a fraction of a systemic dosage.
[0237] Compounds/bioconjugates of the invention can be administered by other controlled- release means or delivery devices that are well known to those of ordinary skill in the art. These include, for example, hydropropylmethyl cellulose, other polymer matrices, gels, permeable membranes, osmotic systems, multilayer coatings, microparticles, liposomes, microspheres, or the like, or a combination of any of the above to provide the desired release profile in varying proportions. Other methods of controlled-release delivery of compounds/bioconjugates will be known to the skilled artisan and are within the scope of the invention,
(iv) Inhalation Administration
[0238] Compounds/bioconjugates of the invention can be administered directly to the lung of a patient/subject by inhalation. For administration by inhalation, a compound/bioconjugate can be conveniently delivered to the lung by a number of different devices. For example, a Metered Dose Inhaler ("MDI") which utilizes canisters that contain a suitable low boiling point propellant, e.g., dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas can be used to deliver a compound/bioconjugate directly to the lung. MDI devices are available from a number of suppliers such as 3M Corporation, Aventis, Boehringer Ingleheim, Forest Laboratories, GlaxoSmithKline, Merck & Co. and Vectura. [0239] Alternatively, a Dry Powder Inhaler (DPI) device can be used to administer a compound/bioconjugate to the lung. DPI devices typically use a mechanism such as a burst of gas to create a cloud of dry powder inside a container, which can then be inhaled by the patient. DPI devices are also well known in the art and can be purchased from a number of vendors which include, for example, GlaxoSmithKline, Nektar Therapeutics, Innovata and Vectura. A popular variation is the multiple dose DPI ("MDDPI") system, which allows for the delivery of more than one therapeutic dose. MDDPI devices are available from companies such as AstraZeneca, GlaxoSmithKline, TEVA, Merck & Co., SkyePharma and Vectura. For example, capsules and cartridges of gelatin for use in an inhaler or insufflator can be formulated containing a powder mix of the compound/bioconjugate and a suitable powder base such as lactose or starch for these systems.
[0240] Another type of device that can be used to deliver a compound/bioconjugate to the lung is a liquid spray device supplied, for example, by Aradigm Corporation. Liquid spray systems use extremely small nozzle holes to aerosolize liquid compound/bioconjugate formulations that can then be directly inhaled into the lung. For example, a nebulizer device can be used to deliver a compound/bioconjugate to the lung. Nebulizers create aerosols from liquid compound/bioconjugate formulations by using, for example, ultrasonic energy to form fine particles that can be readily inhaled. Examples of nebulizers include devices supplied by Aventis and Battelle.
[0241] In another example, an electrohydrodynamic ("EHD") aerosol device can be used to deliver a compound/bioconjugate to the lung. EHD aerosol devices use electrical energy to aerosolize liquid compound/bioconjugate solutions or suspensions. The electrochemical properties of the compound/bioconjugate formulation are important parameters to optimize when delivering this compound/bioconjugate to the lung with an EHD aerosol device. Such optimization is routinely performed by one of skill in the art. Other methods of intra-pulmonary delivery of compounds/bioconjugates will be known to the skilled artisan and are within the scope of the invention.
[0242] Liquid compound/bioconjugate formulations suitable for use with nebulizers and liquid spray devices and EHD aerosol devices will typically include the compound/bioconjugate with a pharmaceutically acceptable carrier. In one exemplary embodiment, the pharmaceutically acceptable carrier is a liquid such as alcohol, water, polyethylene glycol or a perfluorocarbon. Optionally, another material can be added to alter the aerosol properties of the solution or suspension of the compound/bioconjugate. For example, this material can be a liquid such as an alcohol, glycol, polyglycol or a fatty acid. Other methods of formulating liquid compound/bioconjugate solutions or suspensions suitable for use in aerosol devices are known to those of skill in the art.
(v) Depot Administration [0243] A compound/bioconjugate of the invention can be formulated as a depot preparation. Such long-acting formulations can be administered by implantation (e.g., subcutaneously or intramuscularly) or by intramuscular injection. Accordingly, the compound/bioconjugate can be formulated with suitable polymeric or hydrophobic materials such as an emulsion in an acceptable oil or ion exchange resin, or as sparingly soluble derivatives such as a sparingly soluble salt. Other methods of depot delivery of compounds/bioconjugates will be known to the skilled artisan and are within the scope of the invention.
(vi) Topical Administration
[0244] For topical application, a compound/bioconjugate can be combined with a pharmaceutically acceptable carrier so that an effective dosage is delivered, based on the desired activity ranging from an effective dosage, for example, of 1 .0 μΜ to 1 .0 mM. In one aspect of the invention, a topical formulation of a compound/bioconjugate can be applied to the skin. The pharmaceutically acceptable carrier can be in the form of, for example, and not by way of limitation, an ointment, cream, gel, paste, foam, aerosol, suppository, pad or gelled stick.
[0245] A topical formulation can include a therapeutically effective amount of a compound/bioconjugate in an ophthalmologically acceptable excipient such as buffered saline, mineral oil, vegetable oils such as corn or arachis oil, petroleum jelly, Miglyol 182, alcohol solutions, or liposomes or liposome-like products. Any of these formulations of such compounds/bioconjugates can include preservatives, antioxidants, antibiotics, immunosuppressants, and other biologically or pharmaceutically effective agents that do not exert a significant detrimental effect on the compound/bioconjugate. Other methods of topical delivery of compounds/bioconjugates will be known to the skilled artisan and are within the scope of the invention.
(vii) Rectal Administration
[0246] Compounds/bioconjugates of the invention can be formulated in rectal formulations such as suppositories or retention enemas that include conventional suppository bases such as cocoa butter or other glycerides and/or binders and/or carriers such as triglycerides, microcrystalline cellulose, gum tragacanth or gelatin. Rectal formulations can contain a compound/bioconjugate in the range of 0.5% to 10% by weight, for example. Other methods of rectal delivery of compounds/bioconjugates will be known to the skilled artisan and are within the scope of the invention.
(viii) Other Systems of Administration
[0247] Various other delivery systems are known in the art and can be used to administer the compounds/bioconjugates of the invention. Moreover, these and other delivery systems can be combined and/or modified to promote optimization of the administration of compounds/bioconjugates of the present invention. Exemplary formulations that include compounds/bioconjugates of the present invention are described elsewhere herein (the compounds/bioconjugates of the present invention are indicated as the active ingredient, but those of skill in the art will recognize that pro-drugs and compound combinations are also meant to be encompassed by this term).
4d: Formulation
[0248] In an embodiment, the invention provides a medicament which comprises a therapeutically effective amount of one or more compositions of the invention, such as a compound of any one of formulas (FX1 ) - (FX113). In an embodiment, the invention provides a medicament which comprises a diagnostically effective amount of one or more compositions of the invention. In an embodiment, the invention provides a method for making a medicament for treatment of a condition described herein, such as the treatment of cancer, inflammation, stenosis or a vascular disease. In an embodiment, the invention provides a method for making a medicament for diagnosis or aiding in the diagnosis of a condition described herein, such as the diagnosis of cancer, inflammation, stenosis or a vascular disease. In an embodiment, the invention provides the use of one or more compositions set forth herein for the making of a medicament for the treatment of cancer, inflammation, stenosis or a vascular disease. In an embodiment, the invention provides the use of one or more compositions set forth herein for the treatment of a disease. In an embodiment, the invention provides the use of one or more compositions set forth herein for the diagnosis of a disease. Compositions of the invention include formulations and preparations comprising one or more of the present optical agents provided in an aqueous solution, such as a pharmaceutically acceptable formulation or preparation. Optionally, compositions of the invention further comprise one or more pharmaceutically acceptable surfactants, buffers, electrolytes, salts, carriers, binders, coatings, preservatives and/or excipients.
[0249] In an embodiment, the invention provides a pharmaceutical formulation having an active ingredient comprising a composition of the invention, such as a compound of any one of formulas (FX1 ) - (FX113). In an embodiment, the invention provides a method of synthesizing a composition of the invention or a pharmaceutical formulation thereof, such as a compound of any one of formulas (FX1 ) - (FX113). In an embodiment, a pharmaceutical formulation comprises one or more excipients, carriers, diluents, and/or other components as would be understood in the art. Preferably, the components meet the standards of the National Formulary ("IMF"), United States Pharmacopoeia ("USP"; United States Pharmacopeial Convention Inc., Rockville, Maryland), or Handbook of Pharmaceutical Manufacturing Formulations (Sarfaraz K. Niazi, all volumes, ISBN: 9780849317521 , ISBN 10: 0849317525; CRC Press, 2004). See, e.g., United States Pharmacopeia and National Formulary (USP 30-NF 25), Rockville, MD: United States Pharmacopeial Convention (2007 and 2008), and each of any earlier editions; The Handbook of Pharmaceutical Excipients, published jointly by the American Pharmacists Association and the Pharmaceutical Press (Pharmaceutical Press (2005) (ISBN-10: 0853696187, ISBN-13: 978-0853696186)); Merck Index, Merck & Co., Rahway, N.J.; and Gilman et al., (eds) (1996); Goodman and Gilman's: The Pharmacological Bases of Therapeutics, 8th Ed., Pergamon Press. In embodiments, the formulation base of the formulations of the invention comprises physiologically acceptable excipients, namely, at least one binder and optionally other physiologically acceptable excipients. Physiologically acceptable excipients are those known to be usable in the pharmaceutical technology sectors and adjacent areas, particularly, those listed in relevant pharmacopeias (e.g. DAB, Ph. Eur., BP, NF, USP), as well as other excipients whose properties do not impair a physiological use.
[0250] This invention also is directed, in part, to pharmaceutical compositions including a therapeutically effective amount of a compound or salt of this invention, as well as processes for making such compositions. Such compositions generally include one or more pharmaceutically acceptable carriers (e.g., excipients, vehicles, auxiliaries, adjuvants, diluents) and can include other active ingredients. Formulation of these compositions can be achieved by various methods known in the art. A general discussion of these methods can be found in, for example, Hoover, John E., Remington's Pharmaceutical Sciences (Mack Publishing Co., Easton, PA: 1975). See also, Lachman, L, eds., Pharmaceutical Dosage Forms (Marcel Decker, New York, N. Y., 1980).
[0251] The diagnostic and therapeutic formulations of this invention and medicaments of this invention can further comprise one or more pharmaceutically acceptable carriers, excipients, buffers, emulsifiers, surfactants, electrolytes or diluents. Such compositions and medicaments are prepared in accordance with acceptable pharmaceutical procedures, such as, for example, those described in Remingtons Pharmaceutical Sciences, 17th edition, ed. Alfonoso R. Gennaro, Mack Publishing Company, Easton, Pa. (1985).
[0252] Compositions of the invention include formulations and preparations comprising one or more of the present compounds provided in an aqueous solution, such as a pharmaceutically acceptable formulation or preparation. Optionally, compositions of the invention further comprise one or more pharmaceutically acceptable surfactants, buffers, electrolytes, salts, carriers, binders, coatings, preservatives and/or excipients.
[0253] Compounds and bioconjugates of the present invention can be formulated by known methods for administration to a subject using several routes which include, but are not limited to, parenteral, oral, topical, intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal, epidural, and ophthalmic routes. An individual compound/bioconjugate can be administered in combination with one or more additional compounds/bioconjugates of the present invention and/or together with other biologically active or biologically inert agents. Such biologically active or inert agents can be in fluid or mechanical communication with the compound(s)/bioconjugate(s) or attached to the compound(s)/bioconjugate(s) by ionic, covalent, Van der Waals, hydrophobic, hydrophilic or other physical forces. It is preferred that administration is localized in a subject, but administration can also be systemic.
[0254] Compounds and bioconjugates of the present invention can be formulated by any conventional manner using one or more pharmaceutically acceptable carriers. Thus, the compound(s)/bioconjugate(s) and their pharmaceutically acceptable salts and solvates can be specifically formulated for administration, e.g., by inhalation or insufflation (either through the mouth or the nose) or oral, buccal, parenteral or rectal administration. The compounds/bioconjugates can take the form of charged, neutral and/or other pharmaceutically acceptable salt forms. Examples of pharmaceutically acceptable carriers include, but are not limited to, those described in REMINGTON'S PHARMACEUTICAL SCIENCES (A.R. Gennaro, Ed.), 20th edition, Williams & Wilkins PA, USA (2000).
[0255] Compounds and bioconjugates of the present invention can be formulated in the form of solutions, suspensions, emulsions, tablets, pills, capsules, powders, controlled- or sustained- release formulations and the like. Such formulations will contain a therapeutically effective amount of the compound/bioconjugate, preferably in purified form, together with a suitable amount of carrier so as to provide the form for proper administration to the patient. The formulation should suit the mode of administration.
[0256] Pharmaceutically acceptable carriers that can be used in conjunction with the compounds of the invention are well known to those of ordinary skill in the art. Carriers can be selected based on a number of factors including, for example, the particular compound(s) or pharmaceutically acceptable salt(s) used; the compound's concentration, stability, and intended bioavailability; the condition being treated; the subject's age, size, and general condition; the route of administration; etc. A general discussion related to carriers can be found in, for example, J.G. Nairn, Remington's Pharmaceutical Science, pp. 1492-1517 (A. Gennaro, ed., Mack Publishing Co., Easton, Pa. (1985)).
[0257] Solid dosage forms for oral administration include, for example, capsules, tablets, gelcaps, pills, dragees, troches, powders, granules, and lozenges. In such solid dosage forms, the compounds or pharmaceutically acceptable salts thereof can be combined with one or more pharmaceutically acceptable carriers. The compounds and pharmaceutically acceptable salts thereof can be mixed with carriers including, but not limited to, lactose, sucrose, starch powder, corn starch, potato starch, magnesium carbonate, microcrystalline cellulose, cellulose esters of alkanoic acids, cellulose alkyl esters, talc, stearic acid, magnesium stearate, magnesium oxide, sodium and calcium salts of phosphoric and sulfuric acids, sodium carbonate, agar, mannitol, sorbitol, sodium saccharin, gelatin, acacia gum, alginic acid, sodium alginate, tragacanth, colloidal silicon dioxide, croscarmellose sodium, polyvinylpyrrolidone, and/or polyvinyl alcohol, and then tableted or encapsulated for convenient administration. Such capsules or tablets can contain a controlled-release formulation, as can be provided in a dispersion of the compound or salt in hydroxypropylmethyl cellulose. In the case of capsules, tablets, and pills, the dosage forms also can include buffering agents, such as sodium citrate, or magnesium or calcium carbonate or bicarbonate. Tablets and pills additionally can, for example, include a coating (e.g., an enteric coating) to delay disintegration and absorption. The concentration of the present compounds in a solid oral dosage form can be from about 5 to about 50% for example, and in certain aspects from about 8 to about 40%, and in another aspect from about 10 to about 30% by weight based on the total weight of the composition.
[0258] Liquid dosage forms of the compounds of the invention for oral administration include, for example, pharmaceutically acceptable emulsions, solutions, suspensions, syrups, and elixirs containing inert diluents commonly used in the art (e.g., water). Such compositions also can include adjuvants, such as wetting, emulsifying, suspending, flavoring (e.g., sweetening), and/or perfuming agents. The concentration of the present compounds in the liquid dosage form can be from about 0.01 to about 5 mg, and in certain aspects from about 0.01 to about 1 mg, and in another aspect from about 0.01 to about 0.5 mg per ml of the composition. Low concentrations of the compounds of the invention in liquid dosage form can be prepared in the case that the compound is more soluble at low concentrations. Techniques for making oral dosage forms useful in the invention are generally described in, for example, Modern Pharmaceutics, Chapters 9 and 10 (Banker & Rhodes, Editors (1979)). See also, Lieberman et al., Pharmaceutical Dosage Forms: Tablets (1981 ). See also, Ansel, Introduction to Pharmaceutical Dosage Forms (2nd Edition (1976)).
[0259] In some aspects of the invention, tablets or powders for oral administration can be prepared by dissolving the compound in a pharmaceutically acceptable solvent capable of dissolving the compound to form a solution and then evaporating when the solution is dried under vacuum. A carrier can also be added to the solution before drying. The resulting solution can be dried under vacuum to form a glass. The glass can then be mixed with a binder to form a powder. This powder can be mixed with fillers or other conventional tableting agents, and then processed to form a tablet. Alternatively, the powder can be added to a liquid carrier to form a solution, emulsion, suspension, or the like.
[0260] In some aspects, solutions for oral administration are prepared by dissolving the compound in a pharmaceutically acceptable solvent capable of dissolving the compound to form a solution. An appropriate volume of a carrier is added to the solution while stirring to form a pharmaceutically acceptable solution for oral administration.
[0261] In some embodiments, a liposome or micelle can be utilized as a carrier or vehicle for the composition. For example, in some embodiments, the compound can be a part of the lipophilic bilayers or micelle, and the targeting ligand, if present, can be on the external surface of the liposome or micelle. As another example, a targeting ligand can be externally attached to the liposome or micelle after formulation for targeting the liposome or micelle (which contains the optical agents) to the desired tissue, organ, or other site in the body.
[0262] Injectable preparations (e.g., sterile injectable aqueous or oleaginous suspensions) can be formulated according to the known art using suitable dispersing, wetting agents, and/or suspending agents. Acceptable vehicles for parenteral use include both aqueous and nonaqueous pharmaceutically-acceptable solvents. Suitable pharmaceutically acceptable aqueous solvents include, for example, water, saline solutions, dextrose solutions (such as DW5), electrolyte solutions, etc.
[0263] In one embodiment, the present compounds are formulated as nanoparticles or microparticles. Use of such nanoparticle or microparticle formulations can be beneficial for some applications to enhance delivery, localization, target specificity, administration, etc. of the compound. Potentially useful nanoparticles and microparticles include, but are not limited to, micelles, liposomes, microemulsions, nanoemulsions, vesicles, tubular micelles, cylindrical micelles, bilayers, folded sheets structures, globular aggregates, swollen micelles, inclusion complex, encapsulated droplets, microcapsules, nanocapsules or the like. As will be understood by those having skill in the art, the present compounds can be located inside the nanoparticle or microparticle, within a membrane or wall of the nanoparticle or microparticle, or outside of (but bonded to or otherwise associated with) the nanoparticle or microparticle. The agent formulated in nanoparticles or microparticles can be administered by any of the routes previously described. In a formulation applied topically, the compound is slowly released over time. In an injectable formulation, the liposome, micelle, capsule, etc., circulates in the bloodstream and is delivered to the desired site (e.g., target tissue).
[0264] Preparation and loading of nanoparticles and microparticles are well known in the art. As one example, liposomes can be prepared from dipalmitoyi phosphatidylcholine (DPPC) or egg phosphatidylcholine (PC) because this lipid has a low heat transition. Liposomes are made using standard procedures as known to one skilled in the art (e.g., Braun-Falco et al., (Eds.), Griesbach Conference, Liposome Dermatics, Springer-Verlag, Berlin (1992), pp. 69 81 ; 91 1 17. Polycaprolactone, poly(glycolic) acid, poly(lactic) acid, polyanhydride or lipids can be formulated as microspheres. As an illustrative example, the present compounds can be mixed with polyvinyl alcohol (PVA), the mixture then dried and coated with ethylene vinyl acetate, then cooled again with PVA. In a liposome, the present compounds can be within one or both lipid bilayers, in the aqueous between the bilayers, or within the center or core. Liposomes can be modified with other molecules and lipids to form a cationic liposome. Liposomes can also be modified with lipids to render their surface more hydrophilic which increases their circulation time in the bloodstream. The thus-modified liposome has been termed a "stealth" liposome, or a long-lived liposome, as described in U.S. Pat. No. 6,258,378, and in Stealth Liposomes, Lasic and Martin (Eds.) 1995 CRC Press, London. Encapsulation methods include detergent dialysis, freeze drying, film forming, injection, as known to one skilled in the art and disclosed in, for example, U.S. Pat. No. 6,406,713. Optionally, the present compositions and methods include a micelle delivery system, for example, involving one or more PEG-based amphiphilic polymers developed for drug delivery including: PEG-poly(s-caprolactone), PEG-poly(amino acid), PEG- polylactide or PEG-phospholipid constructs; a cross linked poly(acrylic acid) polymer system, a phospholipid-based system and/or block copolymer systems comprising one or more of the following polymer blocks: a poly(lactic acid) polymer block; a poly(propylene glycol) polymer block; a poly(amino acid) polymer block; a poly(ester) polymer block; a poly (ε-caprolactone) polymer block; a poly(ethylene glycol) block, a poly(acrylic acid) block; a polylactide block; a polyester block; a polyamide block; a polyanhydride block; a polyurethane block; a polyimine block; a polyurea block; a polyacetal block; a polysaccharide block; and a polysiloxane block.
[0265] Suitable pharmaceutically-acceptable nonaqueous solvents include, but are not limited to, the following (as well as mixtures thereof):
[0266] (i) Alcohols (these include, for example, o-glycerol formal, β-glycerol formal, 1 , 3- butyleneglycol, aliphatic or aromatic alcohols having from 2 to about 30 carbons (e.g., methanol, ethanol, propanol, isopropanol, butanol, t-butanol, hexanol, octanol, amylene hydrate, benzyl alcohol, glycerin (glycerol), glycol, hexylene, glycol, tetrahydrofuranyl alcohol, cetyl alcohol, and stearyl alcohol), fatty acid esters of fatty alcohols (e.g., polyalkylene glycols, such as polypropylene glycol and polyethylene glycol), sorbitan, sucrose, and cholesterol);
[0267] (ii) Amides, which include, for example, dimethylacetamide (DMA), benzyl benzoate DMA, dimethylformamide, N-hydroxyethyO-lactamide, N, N-dimethylacetamide-amides, 2- pyrrolidinone, 1-methyl-2-pyrrolidinone, and polyvinylpyrrolidone;
[0268] (iii) Esters, which include, for example, acetate esters (e.g., monoacetin, diacetin, and triacetin), aliphatic and aromatic esters (e.g., ethyl caprylate or octanoate, alkyl oleate, benzyl benzoate, or benzyl acetate), dimethylsulfoxide (DMSO), esters of glycerin (e.g., mono, di, and tri-glyceryl citrates and tartrates), ethyl benzoate, ethyl acetate, ethyl carbonate, ethyl lactate, ethyl oleate, fatty acid esters of sorbitan, glyceryl monostearate, glyceride esters (e.g., mono, di, or tri-glycerides), fatty acid esters (e.g., isopropyl myristrate), fatty acid derived PEG esters (e.g., PEG-hydroxyoleate and PEG-hydroxystearate), N-methyl pyrrolidinone, pluronic 60, polyoxyethylene sorbitol oleic polyesters (e.g., poly(ethoxylated)3o-6o sorbitol poly(oleate)2-4, poly(oxyethylene)i5-2o monooleate, poly(oxyethylene)i5-2o mono 12-hydroxystearate, and poly(oxyethylene)i5-2o mono ricinoleate), polyoxyethylene sorbitan esters (e.g., polyoxyethylene- sorbitan monooleate, polyoxyethylene-sorbitan monopalmitate, polyoxyethylene-sorbitan monolaurate, polyoxyethylene-sorbitan monostearate, and POLYSORBATE 20, 40, 60, and 80 (from ICI Americas, Wilmington, DE)), polyvinylpyrrolidone, alkyleneoxy modified fatty acid esters (e.g., polyoxyl 40 hydrogenated castor oil and polyoxyethylated castor oils, such as CREMOPHOR EL solution or CREMOPHOR RH 40 solution), saccharide fatty acid esters (i.e., the condensation product of a monosaccharide (e.g., pentoses, such as, ribose, ribulose, arabinose, xylose, lyxose, and xylulose; hexoses, such as glucose, fructose, galactose, mannose, and sorbose; trioses; tetroses; heptoses; and octoses), disaccharide (e.g., sucrose, maltose, lactose, and trehalose), oligosaccharide, or a mixture thereof with one or more C4-C22 fatty acids (e.g., saturated fatty acids, such as caprylic acid, capric acid, lauric acid, myristic acid, palmitic acid, and stearic acid; and unsaturated fatty acids, such as palmitoleic acid, oleic acid, elaidic acid, erucic acid, and linoleic acid), and steroidal esters; [0269] (iv) Ethers, for example, alkyl, aryl, and cyclic ethers having from 2 to about 30 carbons. Examples include diethyl ether, tetrahydrofuran, dimethyl isosorbide, diethylene glycol monoethyl ether), and glycofurol (tetrahydrofurfuranyl alcohol polyethylene glycol ether);
[0270] (v) Ketones which typically have from about 3 to about 30 carbons. Examples include acetone, methyl ethyl ketone, and methyl isobutyl ketone;
[0271] (vi) Hydrocarbons which are typically aliphatic, cycloaliphatic, or aromatic hydrocarbons having from about 4 to about 30 carbons. Examples include benzene, cyclohexane, dichloromethane, dioxolanes, hexane, n-decane, n-dodecane, n-hexane, sulfolane, tetramethylenesulfone, tetramethylenesulfoxide, toluene, dimethylsulfoxide (DMSO); and tetramethylene sulfoxide;
[0272] (vii) Oils which include, for example, oils of mineral, vegetable, animal, essential, or synthetic origin. These include: mineral oils, such as aliphatic and wax-based hydrocarbons, aromatic hydrocarbons, mixed aliphatic and aromatic based hydrocarbons, and refined paraffin oil; vegetable oils, such as linseed, tung, safflower, soybean, castor, cottonseed, groundnut, rapeseed, coconut, palm, olive, corn, corn germ, sesame, persic, and peanut oil; glycerides, such as mono-, di-, and triglycerides; animal oils, such as fish, marine, sperm, cod-liver, haliver, squaiene, squalane, and shark liver oil; oleic oils; and polyoxyethylated castor oil;
[0273] (viii) Alkyl, alkenyl, or aryl halides which include, for example, alkyl or aryl halides having from 1 to about 30 carbons and one or more halogen substituents. Examples include: methylene chloride; monoethanolamine; petroleum benzin; trolamine; omega-3 polyunsaturated fatty acids (e.g., alpha-linolenic acid, eicosapentaenoic acid, docosapentaenoic acid, or docosahexaenoic acid); polyglycol ester of 12-hydroxystearic acid and polyethylene glycol (SOLUTOL HS-15, from BASF, Ludwigshafen, Germany); polyoxyethylene glycerol; sodium laurate; sodium oleate; and sorbitan monooleate.
[0274] Other pharmaceutically acceptable solvents for use in the invention are well known to those of ordinary skill in the art. General discussion relating to such solvents can be found in, for example, The Chemotherapy Source Book (Williams & Wilkens Publishing), The Handbook of Pharmaceutical Excipients, (American Pharmaceutical Association, Washington, D.C., and The Pharmaceutical Society of Great Britain, London, England, 1968), Modern Pharmaceutics 3d ed., (G. Banker et. al., eds., Marcel Dekker, Inc., New York, New York (1995)), The Pharmacological Basis of Therapeutics, (Goodman & Gilman, McGraw Hill Publishing), Pharmaceutical Dosage Forms, (H. Lieberman et. al., eds., Marcel Dekker, Inc., New York, New York (1980)), Remington's Pharmaceutical Sciences, 19th ed., (A. Gennaro, ed., Mack Publishing, Easton, PA, (1995)), The United States Pharmacopeia 24, The National Formulary 19, (National Publishing, Philadelphia, PA (2000)); Spiegel, A.J., et al., "Use of Nonaqueous Solvents in Parenteral Products," J. Pharma. Sciences, Vol. 52, No. 10, pp. 917-927 (1963).
[0275] Solvents useful in the invention include, but are not limited to, those known to stabilize present compounds or pharmaceutically acceptable salts thereof. These can include, for example, oils rich in triglycerides, such as safflower oil, soybean oil, and mixtures thereof; and alkyleneoxy-modified fatty acid esters, such as polyoxyl 40 hydrogenated castor oil and polyoxyethylated castor oils (e.g., CREMOPHOR EL solution or CREMOPHOR RH 40 solution). Commercially available triglycerides include INTRALI PID emulsified soybean oil (Kabi- Pharmacia Inc., Stockholm, Sweden), NUTRALIPID emulsion (McGaw, Irvine, California), LI POSYN II 20% emulsion (a 20% fat emulsion solution containing 100 mg safflower oil, 100 mg soybean oil, 12 mg egg phosphatides, and 25 mg glycerin per ml of solution; Abbott Laboratories, Chicago, IL), LIPOSYN III 2% emulsion (a 2% fat emulsion solution containing 100 mg safflower oil, 100 mg soybean oil, 12 mg egg phosphatides, and 25 mg glycerin per ml of solution; Abbott Laboratories, Chicago, IL), natural or synthetic glycerol derivatives containing the docosahexaenoyi group at levels of from about 25 to about 100% (by weight based on the total fatty acid content) (DHASCO from Martek Biosciences Corp., Columbia, MD; DHA MAGURO from Daito Enterprises, Los Angeles, CA; SOYACAL; and TRAVEMULSION). Ethanol in particular is a useful solvent for dissolving a compound or pharmaceutically acceptable salt thereof to form solutions, emulsions, and the like.
[0276] Additional components can be included in the compositions of this invention for various purposes generally known in the pharmaceutical industry. These components tend to impart properties that, for example, enhance retention of the present compounds or salt thereof at the site of administration, protect the stability of the composition, control the pH, and facilitate processing of the compound or salt thereof into pharmaceutical formulations, and the like. Specific examples of such components include cryoprotective agents; agents for preventing reprecipitation of the compound or salt surface; active, wetting, or emulsifying agents (e.g., lecithin, polysorbate-80, TWEEN 80, pluronic 60, and polyoxyethylene stearate); preservatives (e.g., ethyl-p-hydroxybenzoate); microbial preservatives (e.g., benzyl alcohol, phenol, m-cresol, chlorobutanol, sorbic acid, thimerosal, and paraben); agents for adjusting pH or buffering agents (e.g., acids, bases, sodium acetate, sorbitan monolaurate, etc.); agents for adjusting osmolarity (e.g., glycerin); thickeners (e.g., aluminum monostearate, stearic acid, cetyl alcohol, stearyl alcohol, guar gum, methyl cellulose, hydroxypropylcellulose, tristearin, cetyl wax esters, polyethylene glycol, etc.); colorants; dyes; flow aids; non-volatile silicones (e.g., cyclomethicone); clays (e.g., bentonites); adhesives; bulking agents; flavorings; sweeteners; adsorbents; fillers (e.g., sugars such as lactose, sucrose, mannitol, sorbitol, cellulose, calcium phosphate, etc.); diluents (e.g., water, saline, electrolyte solutions, etc.); binders (e.g., gelatin; gum tragacanth; methyl cellulose; hydroxypropyl methylcellulose; sodium carboxymethyl cellulose; polyvinylpyrrolidone; sugars; polymers; acacia; starches, such as maize starch, wheat starch, rice starch, and potato starch; etc.); disintegrating agents (e.g., starches, such as maize starch, wheat starch, rice starch, potato starch, and carboxymethyl starch; cross-linked polyvinyl pyrrolidone; agar; alginic acid or a salt thereof, such as sodium alginate; croscarmellose sodium; crospovidone; etc); lubricants (e.g., silica; talc; stearic acid and salts thereof, such as magnesium stearate; polyethylene glycol; etc.); coating agents (e.g., concentrated sugar solutions including gum arabic, talc, polyvinyl pyrrolidone, carbopol gel, polyethylene glycol, titanium dioxide, etc.); and antioxidants (e.g., sodium metabisulfite, sodium bisulfite, sodium sulfite, dextrose, phenols, thiophenols, etc.).
[0277] Techniques and compositions for making parenteral dosage forms are generally known in the art. Formulations for parenteral administration can be prepared from one or more sterile powders and/or granules having a compound or salt of this invention and one or more of the carriers or diluents mentioned for use in the formulations for oral administration. The powder or granule typically is added to an appropriate volume of a solvent (typically while agitating (e.g., stirring) the solvent) that is capable of dissolving the powder or granule. Particular solvents useful in the invention include, for example, water, polyethylene glycol, propylene glycol, ethanol, corn oil, cottonseed oil, peanut oil, sesame oil, benzyl alcohol, sodium chloride, and/or various buffers.
[0278] Emulsions for parenteral administration can be prepared by, for example, dissolving a compound or salt of this invention in any pharmaceutically acceptable solvent capable of dissolving the compound to form a solution; and adding an appropriate volume of a carrier to the solution while stirring to form the emulsion. Solutions for parenteral administration can be prepared by, for example, dissolving a compound or salt of this invention in any pharmaceutically acceptable solvent capable of dissolving the compound to form a solution; and adding an appropriate volume of a carrier to the solution while stirring to form the solution.
[0279] Suppositories for rectal administration can be prepared by, for example, mixing the drug with a suitable nonirritating excipient that is solid at ordinary temperatures, but liquid at the rectal temperature and will therefore melt in the rectum to release the drug. Suitable excipients include, for example, cocoa butter; synthetic mono-, di-, or triglycerides; fatty acids; and/or polyethylene glycols.
[0280] Every formulation or combination of components described or exemplified herein can be used to practice the invention, unless otherwise stated,
(i) Binding Agents
[0281] Binding agents include, but are not limited to, corn starch, potato starch, or other starches, gelatin, natural and synthetic gums such as acacia, sodium alginate, alginic acid, other alginates, powdered tragacanth, guar gum, cellulose and its derivatives (e.g., ethyl cellulose, cellulose acetate, carboxymethyl cellulose calcium, sodium carboxymethyl cellulose), polyvinyl pyrrolidone, methyl cellulose, pre-gelatinized starch, hydroxypropyl methyl cellulose, (e.g., Nos. 2208, 2906, 2910), microcrystalline cellulose, and mixtures thereof. Suitable forms of microcrystalline cellulose include, for example, the materials sold as AVICEL-PH-101 , AVICEL- PH-103 and AVICEL-PH-105 (available from FMC Corporation, American Viscose Division, Avicel Sales, Marcus Hook, Pennsylvania, USA). An exemplary suitable binder is a mixture of microcrystalline cellulose and sodium carboxymethyl cellulose sold as AVICEL RC-581 by FMC Corporation.
(ii) Fillers
[0282] Fillers include, but are not limited to, talc, calcium carbonate (e.g., granules or powder), lactose, microcrystalline cellulose, powdered cellulose, dextrates, kaolin, mannitol, silicic acid, sorbitol, starch, pre-gelatinized starch, and mixtures thereof.
(iii) Lubricants
[0283] Lubricants include, but are not limited to, calcium stearate, magnesium stearate, mineral oil, electromagnetic radiation mineral oil, glycerin, sorbitol, mannitol, polyethylene glycol, other glycols, stearic acid, sodium lauryl sulfate, talc, hydrogenated vegetable oil (e.g., peanut oil, cottonseed oil, sunflower oil, sesame oil, olive oil, corn oil, and soybean oil), zinc stearate, ethyl oleate, ethyl laurate, agar, and mixtures thereof. Additional lubricants include, for example, a syloid silica gel (AEROSIL 200, manufactured by W.R. Grace Co. of Baltimore, Maryland, USA), a coagulated aerosol of synthetic silica (marketed by Deaussa Co. of Piano, Texas, USA), CAB-O-SIL (a pyrogenic silicon dioxide product sold by Cabot Co. of Boston, Massachusetts, USA), and mixtures thereof.
(iv) Disintegrants
[0284] Disintegrants include, but are not limited to, agar-agar, alginic acid, calcium carbonate, microcrystalline cellulose, croscarmellose sodium, crospovidone, polacrilin potassium, sodium starch glycolate, potato or tapioca starch, other starches, pre-gelatinized starch, other starches, clays, other algins, other celluloses, gums, and mixtures thereof.
[0285] Tablets or capsules can optionally be coated by methods well known in the art. If binders and/or fillers are used with a compound/bioconjugate of the invention, they are typically formulated as about 50 to about 99 weight percent of the compound/bioconjugate. In one aspect, about 0.5 to about 15 weight percent of disintegrant, and particularly about 1 to about 5 weight percent of disintegrant, can be used in combination with the compound. A lubricant can optionally be added, typically in an amount of less than about 1 weight percent of the compound/bioconjugate. Techniques and pharmaceutically acceptable additives for making solid oral dosage forms are described in Marshall, SOLID ORAL DOSAGE FORMS, Modern Pharmaceutics (Banker and Rhodes, Eds.), 7:359-427 (1979). Other formulations are known in the art.
[0286] Liquid preparations for oral administration can take the form of solutions, syrups or suspensions. Alternatively, the liquid preparations can be presented as a dry product for constitution with water or other suitable vehicle before use. Such liquid preparations can be prepared by conventional means with pharmaceutically acceptable additives such as suspending agents (e.g., sorbitol syrup, cellulose derivatives or hydrogenated edible fats); emulsifying agents (e.g., lecithin or acacia); non-aqueous vehicles (e.g., almond oil, oily esters, ethyl alcohol or fractionated vegetable oils); and/or preservatives (e.g., methyl or propyl-p- hydroxybenzoates or sorbic acid). The preparations can also contain buffer salts, flavoring, coloring, perfuming and sweetening agents as appropriate. Preparations for oral administration can also be formulated to achieve controlled release of the compound/bioconjugate. Oral formulations preferably contain 10% to 95% compound/bioconjugate. In addition, a compound/bioconjugate of the present invention can be formulated for buccal administration in the form of tablets or lozenges formulated in a conventional manner. Other methods of oral delivery of compounds/bioconjugates of the invention will be known to the skilled artisan and are within the scope of the invention.
Formulation 1
[0287] Hard gelatin capsules are prepared using the following ingredients:
TABLE F1
Ingredients (mg/capsule)
Active Ingredient 250.0
Starch 305.0
Magnesium stearate 5.0
[0288] The above ingredients are mixed and filled into hard gelatin capsules in 560 mg quantities.
Formulation 2
[0289] A tablet formula is prepared using the following ingredients:
TABLE F2
Ingredients (mg/tablet)
Active Ingredient 250.0
Cellulose, microcrystalline 400.0
Colloidal silicon dioxide 10.0
Stearic acid 5.0
[0290] The components are blended and compressed to form tablets, each weighing 665 mg. Formulation 3
[0291] A dry powder inhaler formulation is prepared containing the following components: TABLE F3
Ingredients Weight %
Active ingredient 5
Lactose 95
[0292] The active ingredient is mixed with the lactose and the mixture is added to a dry powder inhaling appliance.
Formulation 4
[0293] Tablets, each containing 60 mg of active ingredient, are prepared as follows:
TABLE F4
Ingredients milligrams
Active ingredient 60.0
Starch 45.0
Microcrystalline cellulose 35.0
Polyvinylpyrrolidone (as 10% solution in water) 4.0
Sodium carboxymethyl starch 4.5
Magnesium stearate 0.5
Talc 1.0
Total 150.0
[0294] The active ingredient, starch and cellulose are passed through a No. 20 mesh U.S. sieve and mixed thoroughly. The solution of polyvinylpyrrolidone is mixed with the resultant powders which are then passed through a 16 mesh U.S. sieve. The granules as produced are dried at 50-60 °C and passed through a 16 mesh U.S. sieve. The sodium carboxymethyl starch, magnesium stearate, and talc, previously passed through a No. 30 mesh U.S. sieve, are then added to the granules which, after mixing, are compressed on a tablet machine to yield tablets each weighing 150 mg.
Formulation 5
[0295] Capsules, each containing 80 mg of active ingredient are made as follows: TABLE F5
Ingredients milligrams
Active ingredient 80.0
Starch 109.0
Magnesium stearate 1 .0
Total 190.0
[0296] The active ingredient, cellulose, starch, and magnesium stearate are blended, passed through a No. 20 mesh U.S. sieve, and filled into hard gelatin capsules in 190 mg quantities. Formulation 6
[0297] Suppositories, each containing 225 mg of active ingredient, are made as follows:
TABLE F6
Ingredients milligrams
Active Ingredient 225
Saturated fatty acid glycerides to 2000
[0298] The active ingredient is passed through a No. 60 mesh U.S. sieve and suspended in the saturated fatty acid glycerides previously melted using the minimum heat necessary. The mixture is then poured into a suppository mold of nominal 2.0 g capacity and allowed to cool.
[0299] Formulation 7
[0300] Suspensions, each containing 50 mg of active ingredient per 5.0 ml dose are made as follows:
TABLE F7
Ingredients milligrams
Active ingredient 50.0 mg
Xanthan gum 4.0 mg
Sodium carboxymethyl cellulose (1 1 %)
Microcrystalline cellulose (89%) 50.0 mg
Sucrose 1 .75 g
Sodium benzoate 10.0 mg
Flavor q.v.
Figure imgf000119_0001
[0301] The active ingredient, sucrose and xantham gum are blended, passed through a No. 10 mesh U.S. sieve, and mixed with a previously made solution of the microcrystalline cellulose and sodium carboxymethyl cellulose in water. The sodium benzoate, flavor, and color are diluted with some of the water and added with stirring. Sufficient water is then added to produce the required volume.
Formulation 8
[0302] Capsules, each containing 150 mg of active ingredient, are made as follows:
TABLE F8
Ingredients milligrams
Active ingredient 150.0
Starch 407.0
Magnesium stearate 3.0
Total 560.0
[0303] The active ingredient, cellulose, starch, and magnesium stearate are blended, passed through a No. 20 mesh U.S. sieve, and filled into hard gelatin capsules in 560 mg quantities.
3f: Kits
[0304] Various embodiments of the present invention include kits. Such kits can include a compound/bioconjugate of the present invention, optionally one or more ingredients for preparing a pharmaceutically acceptable formulation of the compound/bioconjugate, and instructions for use (e.g., administration). When supplied as a kit, different components of a compound/bioconjugate formulation can be packaged in separate containers and admixed immediately before use. Such packaging of the components separately can, if desired, be presented in a pack or dispenser device which can contain one or more unit dosage forms containing the compound/bioconjugate. The pack can, for example, comprise metal or plastic foil such as a blister pack. Such packaging of the components separately can also, in certain instances, permit long-term storage without losing activity of the components. In addition, if more than one route of administration is intended or more than one schedule for administration is intended, the different components can be packaged separately and not mixed prior to use. In various embodiments, the different components can be packaged in one combination for administration together. [0305] It is further contemplated that the compounds and salts of this invention can be used in the form of a kit that is suitable for use in performing the methods described herein, packaged in a container. The kit can contain the compound or compounds and, optionally, appropriate diluents, devices or device components suitable for administration and instructions for use in accordance with the methods of the invention. The devices can include parenteral injection devices, such as syringes or transdermal patch or the like. Device components can include cartridges for use in injection devices and the like. In one aspect, the kit includes a first dosage form including a compound or salt of this invention and a second dosage form including another active ingredient in quantities sufficient to carry out the methods of the invention. The first dosage form and the second dosage form together can include a therapeutically effective amount of the compounds for treating the targeted condition(s).
[0306] In certain embodiments, kits can be supplied with instructional materials. Instructions can be printed on paper or other substrate, and/or can be supplied as an electronic-readable medium, such as a floppy disc, mini-CD-ROM, CD-ROM, DVD-ROM, Zip disc, videotape, audio tape, and the like. Detailed instructions can not be physically associated with the kit; instead, a user can be directed to an Internet web site specified by the manufacturer or distributor of the kit, or supplied as electronic mail.
[0307] If desired, the emulsions or solutions described above for oral or parenteral administration can be packaged in IV bags, vials, or other conventional containers in concentrated form, and then diluted with a pharmaceutically acceptable liquid (e.g., saline) to form an acceptable compound concentration before use.
[0308] Kits can include reagents in separate containers such as, for example, sterile water or saline to be added to a lyophilized active component packaged separately. For example, sealed glass ampules can contain lyophilized superoxide dismutase mimetics and in a separate ampule, sterile water, sterile saline or sterile each of which has been packaged under a neutral non-reacting gas, such as nitrogen. Ampules can consist of any suitable material, such as glass, organic polymers, such as polycarbonate, polystyrene, ceramic, metal or any other material typically employed to hold reagents. Other examples of suitable containers include bottles that can be fabricated from similar substances as ampules, and envelopes that can consist of foil- lined interiors, such as aluminum or an alloy. Other containers include test tubes, vials, flasks, bottles, syringes, and the like. Containers can have a sterile access port, such as a bottle having a stopper that can be pierced by a hypodermic injection needle. Other containers can have two compartments that are separated by a readily removable membrane that upon removal permits the components to mix. Removable membranes can be glass, plastic, rubber, and the like.
STATEMENTS REGARDING INCORPORATION BY REFERENCE AND VARIATIONS
[0309] All references throughout this application, for example patent documents including issued or granted patents or equivalents; patent application publications; and non-patent literature documents or other source material; are hereby incorporated by reference herein in their entireties, as though individually incorporated by reference, to the extent each reference is at least partially not inconsistent with the disclosure in this application (for example, a reference that is partially inconsistent is incorporated by reference except for the partially inconsistent portion of the reference).
[0310] The terms and expressions which have been employed herein are used as terms of description and not of limitation, and there is no intention in the use of such terms and expressions of excluding any equivalents of the features shown and described or portions thereof, but it is recognized that various modifications are possible within the scope of the invention claimed. Thus, it should be understood that although the invention has been specifically disclosed by preferred embodiments, exemplary embodiments and optional features, modification and variation of the concepts herein disclosed can be resorted to by those skilled in the art, and that such modifications and variations are considered to be within the scope of this invention as defined by the appended claims. The specific embodiments provided herein are examples of useful embodiments of the invention and it will be apparent to one skilled in the art that the invention can be carried out using a large number of variations of the devices, device components, methods steps set forth in the present description. As will be apparent to one of skill in the art, methods and devices useful for the present methods can include a large number of optional composition and processing elements and steps.
[0311] When a group of substituents is disclosed herein, it is understood that all individual members of that group and all subgroups, including any isomers, enantiomers, and
diastereomers of the group members, are disclosed separately. When a Markush group or other grouping is used herein, all individual members of the group and all combinations and subcombinations possible of the group are intended to be individually included in the disclosure. When a compound is described herein such that a particular isomer, enantiomer or
diastereomer of the compound is not specified, for example, in a formula or in a chemical name, that description is intended to include each isomers and enantiomer of the compound described individual or in any combination. Additionally, unless otherwise specified, all isotopic variants of compounds disclosed herein are intended to be encompassed by the disclosure. For example, it will be understood that any one or more hydrogens in a molecule disclosed can be replaced with deuterium or tritium. Isotopic variants of a molecule are generally useful as standards in assays for the molecule and in chemical and biological research related to the molecule or its use. Methods for making such isotopic variants are known in the art. Specific names of compounds are intended to be exemplary, as it is known that one of ordinary skill in the art can name the same compounds differently.
[0312] It must be noted that as used herein and in the appended claims, the singular forms "a", "an", and "the" include plural reference unless the context clearly dictates otherwise. Thus, for example, reference to "a cell" includes a plurality of such cells and equivalents thereof known to those skilled in the art, and so forth. As well, the terms "a" (or "an"), "one or more" and "at least one" can be used interchangeably herein. It is also to be noted that the terms
"comprising", "including", and "having" can be used interchangeably. The expression "of any of claims XX- YY" (wherein XX and YY refer to claim numbers) is intended to provide a multiple dependent claim in the alternative form, and in some embodiments is interchangeable with the expression "as in any one of claims XX- YY."
[0313] Unless defined otherwise, all technical and scientific terms used herein have the same meanings as commonly understood by one of ordinary skill in the art to which this invention belongs. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, the preferred methods and materials are now described. Nothing herein is to be construed as an admission that the invention is not entitled to antedate such disclosure by virtue of prior invention.
[0314] Whenever a range is given in the specification, for example, a range of integers, a temperature range, a time range, a composition range, wavelength range, power range, energy range, or concentration range, all intermediate ranges and subranges, as well as all individual values included in the ranges given are intended to be included in the disclosure. As used herein, ranges specifically include the values provided as endpoint values of the range. As used herein, ranges specifically include all the integer values of the range. For example, a range of 1 to 100 specifically includes the end point values of 1 and 100. It will be understood that any subranges or individual values in a range or subrange that are included in the description herein can be excluded from the claims herein.
[0315] As used herein, "comprising" is synonymous and can be used interchangeably with "including," "containing," or "characterized by," and is inclusive or open-ended and does not exclude additional, unrecited elements or method steps. As used herein, "consisting of" excludes any element, step, or ingredient not specified in the claim element. As used herein, "consisting essentially of" does not exclude materials or steps that do not materially affect the basic and novel characteristics of the claim. In each instance herein any of the terms "comprising", "consisting essentially of" and "consisting of" can be replaced with either of the other two terms. The invention illustratively described herein suitably can be practiced in the absence of any element or elements, limitation or limitations which is not specifically disclosed herein.
[0316] One of ordinary skill in the art will appreciate that starting materials, biological materials, reagents, synthetic methods, purification methods, analytical methods, assay methods, and biological methods other than those specifically exemplified can be employed in the practice of the invention without resort to undue experimentation. All art-known functional equivalents, of any such materials and methods are intended to be included in this invention. The terms and expressions which have been employed are used as terms of description and not of limitation, and there is no intention that in the use of such terms and expressions of excluding any equivalents of the features shown and described or portions thereof, but it is recognized that various modifications are possible within the scope of the invention claimed. Thus, it should be understood that although the invention has been specifically disclosed by preferred embodiments and optional features, modification and variation of the concepts herein disclosed can be resorted to by those skilled in the art, and that such modifications and variations are considered to be within the scope of this invention as defined by the appended claims.

Claims

What is claimed is:
1. A compound being of the formula (FX1 ) or (FX2):
Figure imgf000124_0001
(FX2) wherein:
each of L1, L2, and L3, if present, is independently CrC10 alkylene, C3-C10 cycloalkylene, C2-C10 alkenylene, C3-C10 cycloalkenylene, C2-C10 alkynylene, ethenylene, ethynylene, phenylene, -CO-, -0-, -S-, -(CH2CH20)a-, -CO(CH2CH20)a- -(CHOH)b- or -CO(CHOH)b-;
each of W1, W2, and W3 is independently a single bond, -(CH2)n- -(HCCH)n- - (CH2CH20)a-, -0-, -S-, -SO-, -S02-, -SO3-, -OS02-, -NR11-, -CO-, -COO-, - OCO-, -OCOO-, -CONR12-, -NR13CO-, -OCONR14-, -NR15COO- -NR16CONR17-, - NR18CSNR19-, -(CH2)mO(CH2)n- -(CH2)mS(CH2)n-,-(CH2)mSO(CH2)n, -(CH2)m
S02(CH2)n- -(CH2)mS03(CH2)n- -(CH2)mOS02(CH2)n- -(CH2)mNR20(CH2)n- - (CH2)mCO(CH2)n- -(CH2)mCOO(CH2)n- -(CH2)mOCO(CH2)n- -(CH2)mOCOO(CH2)n- - (CH2)mCONR21(CH2)n-,-(CH2)mNR22CO(CH2)n- -(CH2)mOCONR23(CH2)n- - (CH2)mNR24COO(CH2)n- -(CH2)mNR25CONR26(CH2)n- -(CH2)mNR27CSNR28(CH2)n- - (CH2)mO(CH2)nNR29CO(CH2)n- -(CH2)mCO(CH2)n(CH2OCH2)a(CH2)nNR30(CH2)nNR31CO- -, -CO(CH2)nNR32CO- or -CONR^CH^N 34-;
each ring Z is independently
Figure imgf000124_0002
each U is independently -0-, -S-, -CR8R9- or -NR10-;
each Y is independently -N- or -CR2-;
each A is independently a group corresponding to an optical dye; each of R1 to R5 and R8 to R10 is independently hydrogen, -OCF3, C C6 alkyl, C3-C6 cycloalkyl, C5-C10 aryl, C5-C10 heteroaryl, CrC6 acyl, C2-C6 alkenyl, C2-C6 alkynyl, C5-C10 alkylaryl, halo, halomethyl, dihalomethyl, trihalomethyl, -CN, -C02R6°, - CONR61 R62, -COR63, -NO2, -SOR64, -OSR65 , -S02R66, -S02OR67, -S02NR68R69, - PO3R70R71 , -OR72, -SR73, -NR74R75, -NR76COR77, -CH2(CHOH)cR78,-(CH2CH20)cR79, PS1 or PS2;
each of R11 to R34 and R60 to R79 is independently hydrogen, C C6 alkyl, C3-C6 cycloalkyl, C5-C10 aryl or C5-C10 heteroaryl;
each PS1 is independently an azide, azo, diazo, oxaza, diaza, dithia, thioxa, or dioxa group;
each PS2 is independently a group corresponding to a porphyrin, benzoporphyrin, phthalocyanine, phenothiazine, chlorin, bacteriochlorin, phthalocyanine, porphyrin, purpurin, merocyanine, pheophorbides, psoralen, aminolevulinic acid, hematoporphyrin derivative, porphycenes, or porphacyanine;
each c is independently 0 or an integer selected from the range of 1 to 100;
each of a and b is independently an integer selected from the range of 1 to 100; each of m and n is independently 0 or an integer selected from the range of 1 to
10; and
each of q, r and x is independently 0 or 1.
2. The compound of claim 1 being of the formula (FX3) or (FX4):
Figure imgf000125_0001
3. The compound of any of claims 1-2, wherein A is a group corresponding to a pyrazine, a cyanine, an indocyanine, a phthalocyanine, a rhodamine, a thiazole, a phenylxanthene, a phenothiazine, a phenoselenazine, a squaraine, a dipyrrolo pyrimidone, an anthraquinone, a tetracene, a quinoline, an acridine, an acridone, a phenanthridine, an azo dye, a phenoxazine, an azulene, an azaazulene, a triphenyl methane dye, an indole, a benzoindole, an
indocarbocyanine, a Nile Red dye, a thionin dye, an isosulfan blue dye, or a
benzoindocarbocyanine.
4. The compound of any of claims 1-3 being of the formula (FX5), (FX6), (FX7) or (FX8):
Figure imgf000126_0001
. The compound of any of claims 1-4 being of the formula (FX9), (FX10), (FX11 ) or (FX12):
);
Figure imgf000127_0001
6. The compound of any of claims 1-5 being of the formula (FX13) or (FX14):
Figure imgf000127_0002
Figure imgf000128_0001
wherein B is -O- or -S-.
7. The compound of any of claims 1-6 being of the formula (FX15) or (FX16):
Figure imgf000128_0002
8. The compound of claim 1 being of the formula (FX17), (FX18) or (FX19):
Figure imgf000128_0003
Figure imgf000129_0001
(FX19).
9. The compound of claim 1 being of the formula (FX20), (FX21 ), (FX22) or (FX23):
Figure imgf000129_0002
wherein B is -O- or -S-.
10. The compound of claim 1 being of the formula (FX24), (FX25), (FX26) or (FX27):
Figure imgf000129_0003
(FX24);
Figure imgf000130_0001
wherein B is -O- or -S-.
1 1. The compound of any of claims 1-10, wherein A is a group corresponding to a pyrazine.
12. The compound of claim 1 1 being of the formula (FX28) or (FX29):
Figure imgf000131_0001
(FX29); wherein
each of X1 , X2 and X3 is independently hydrogen, -OCF3, CrC6 alkyl, C3-C6 cycloalkyi, C5-C10 aryl, C5-C10 heteroaryl, CrC6 acyl, C2-C6 alkenyl, C2-C6 alkynyl, C5-C10 alkylaryl, halo, halomethyl, dihalomethyl, trihalomethyl, -C02R4°, -SOR41 , -OSR42, -S02OR43, - CH2(CH2OCH2)cCH2OH , -POaR^R45, -OR46, -SR47, -NR48R49, -NR50COR51 , -CN , - CON R52R53, -COR54, -NO2, -S02R55, -S02N R56R57, PS1 or PS2; and
each of R40 to R57 is independently hydrogen, CrC6 alkyl, C3-C6 cycloalkyi, C5-C10 aryl, C5-C10 heteroaryl, or pyrrolyl. g of the formula (FX30), (FX31 ), (FX32) or (FX33):
Figure imgf000131_0002
(FX30);
Figure imgf000132_0001
(FX31);
Figure imgf000132_0002
14. The compound of any of claims 11 -13 being of the formula (FX34), (FX35), (FX36) or (FX37):
Figure imgf000132_0003
(FX34);
Figure imgf000133_0001
15. The compound of any of claims 11 -14 being of the formula (FX38) or (FX39):
Figure imgf000133_0002
(FX38); or
Figure imgf000134_0001
wherein B is -O- or -S-.
16. The compound of any of claims 11 -15 being of the formula (FX40) or (FX41 ):
Figure imgf000134_0002
The compound of claim 11 or 12 being of the formula (FX42), (FX43) or (FX44):
Figure imgf000134_0003
Figure imgf000135_0001
18. The compound of claim 1 1 or 12 being of the formula (FX45), (FX46), (FX47) or (FX48):
Figure imgf000135_0002
wherein B is-O- or -S-.
9. The compound of claim 11 or 12 being of the formula (FX49), (FX50), (FX51) or (FX52):
Figure imgf000136_0001
FX49);
Figure imgf000136_0002
20. The compound of claim 11 or 12 being of the formula (FX53), (FX54), (FX55) or (FX56):
Figure imgf000137_0001
(FX53);
Figure imgf000137_0002
(FX54);
Figure imgf000137_0003
(FX55); or
Figure imgf000138_0001
wherein B is-O- or -S-.
21. The compound of claim 1 1 or 12 being of the formula (FX57), (FX58), or (FX59):
Figure imgf000138_0002
The compound of claim 1 1 or 12 being of the formula (FX60), (FX61 ), or (FX62):
Figure imgf000139_0001
23. The compound of any of claims 1-10, wherein A is a group corresponding to an indocyanine.
24. The compound of claim 23 being of the formula (FX63) or (FX64):
Figure imgf000140_0001
(FX64); wherein:
each of X1 to X17 is independently hydrogen, -OCF3, CrC6 alkyl, C3-C6 cycloalkyi, C5- Cio aryl, C5-C10 heteroaryl, CrC6 acyl, C2-C6 alkenyl, C2-C6 alkynyl, C5-C10 alkylaryl, halo, halomethyl, dihalomethyl, trihalomethyl, -C02R4°, -SOR41 , -OSR42, -S02OR43, - CH2(CH2OCH2)cCH2OH , -POaR^R45, -OR46, -SR47, -NR48R49, -NR50COR51 , -CN , - CON R52R53, -COR54, -N02, -S02R55, -S02N R56R57, PS1 or PS2;
each of R40 to R57 is independently hydrogen, C C6 alkyl, C3-C6 cycloalkyi, C5-C10 aryl or C5-C10 heteroaryl;
each of h and g is independently an integer selected from the range of from 1 to 3; and each of j and i is independently 0 or an integer selected from the range of from 1 to 10.
5. The compound of claim 23 or 24 being of the formula (FX65), (FX66), (FX67) or (FX68):
Figure imgf000141_0001
Figure imgf000141_0002
Figure imgf000142_0001
26. The compound of any of claims 23 - 25 being of the formula (FX69), (FX70), (FX71) or (FX72):
Figure imgf000143_0001
141
Figure imgf000144_0001
27. The compound of claim 23 or 24 being of the formula (FX73) or (FX74):
Figure imgf000145_0001
28. The compound of any of claims 1 -10, wherein A is a group corresponding to azulene or azaazulene.
29. The compound of claim 28 being of formula (FX75), (FX76), (FX77) or (FX78):
6),
Figure imgf000146_0001
wherein:
G1 is -N-, -C(K)- or-C(X1)-; G2 is -N-, -C(K)-, or -C(X2)-
G3 is -N-, -C(K)-, or -C(X3)-
G4 is -N-, -C(K)-, or -C(X4)-
G5 is -N-, -C(K)-, or -C(X5)-
G6 is -N-, -C(K)-, or -C(X6)-
G7 is -N-, -C(K)-, or -C(X7)-
G8 is -N-, -C(K)-, or -C(X8)- wherein at most one of G1 - G8 is -N-; and wherein only one of G1 to G8 is -
C(K)-;
each K is a single bond to W1, W2, or W3;
each of X1 to X8 is independently hydrogen, -OCF3, C C6 alkyl, C3-C6 cycloalkyi, C5-C10 aryl, C5-C10 heteroaryl, C C6 acyl, C2-C6 alkenyl, C2-C6 alkynyl, C5-C10 alkylaryl, halo, halomethyl, dihalomethyl, trihalomethyl, -C02R4°, -SOR41, -OSR42, -S02OR43, - CH2(CH2OCH2)cCH2OH, -POaR^R45, -OR46, -SR47, -NR48R49, -NR50COR51, -CN, - CONR52R53, -COR54, -NO2, -S02R55, -S02NR56R57, PS1 or PS2; and
each of R40 to R57 is independently hydrogen, CrC6 alkyl, C3-C6 cycloalkyi, C5-C10 aryl or C5-C10 heteroaryl.
30. The compound of claim 28 or 29, being of the formula (FX79) or (FX80):
Figure imgf000147_0001
The compound of claim 28 or 29, being of the formula (FX81 ) or (FX82):
Figure imgf000148_0001
(FX81 ) or
Figure imgf000148_0002
The compound of claim 28 or 29, being of the formula (FX83) or (FX84):
Figure imgf000149_0001
The compound of claim 28 or 29, being of the formula (FX85), (FX86), (FX87) or (FX88):
Figure imgf000149_0002
(FX85),
Figure imgf000150_0001
The compound of claim 28 or 29, being of the formula (FX89), (FX90), (FX91 ) or (FX92):
Figure imgf000150_0002
(FX89), ),
Figure imgf000151_0001
The compound of claim 28 or 29, being of the formula (FX93), (FX94), (FX95) or (FX96):
Figure imgf000151_0002
(FX93),
Figure imgf000152_0001
36. The compound of claim 28 or 29, being of the formula (FX97), (FX98), (FX99) or (FX100):
Figure imgf000153_0001
37. The compound of claim 28 or 29, being of the formula (FX101 ), (FX102), (FX103) or (FX104
Figure imgf000154_0001
Figure imgf000155_0001
38. The compound of any of claims 1-8, 1 1-17, and 23 - 37 wherein at least one of W1 and L1,
W2 and L2, or W3 and L3 combine to form:
-(CH2)n- -(CH2)mNHCO- -CONH(CH2)nCO- , -0(CH2)n- or
Figure imgf000155_0002
39. The compound of any of claims 1-8, 1 1-17, and 23 - 37, wherein at least one of W1 and L1, W2 and L2 or W3 and L3 combine to form:
Figure imgf000155_0003
Figure imgf000156_0001
40. The compound of any of claims 12-18, and 20, wherein at least one of X1 to X3 is CrC6 alkyl, -OR46, -SR47, -NR48R49, or -NR50COR51; and wherein at least one of X1 to X3 is - CN, -C02R40, -SO2OR43, -CONR52R53, -COR54, -N02, -SOR41, -S02R55, -P03R44R45, halo, Ci-C6 acyl, trihalomethyl, or -S02NR56R57.
41 . The compound of any of claims 12-18, and 20, wherein at least one of X1 to X3 is -OR46, -SR47, -NR48R49, or -NR50COR51.
42. The compound of any of claims 12-18, and 20, wherein at least one of X1 to X3 is - NR48R49, or -NR50COR51.
43. The compound of any of claims 12-18, and 20, wherein at least one of X1 to X3 is -CN, - COzR^ -COR54, -NO2, -SO2R55, or -S02NR56R57.
44. The compound of any of claims 12-18, and 20, wherein at least one of X1 to X3 is - C02R40, -COR54, -S02NR56R57 or -S02R55.
45. The compound of any of claims 12-18, and 20, wherein at least one of X1 to X3 is - NR48R49, and wherein at least one of X1 to X17 is -CO2R40,-COR54, -S02NR56R57 or - S02R55.
46. The compound of any of claims 12-18, and 20, wherein at least one of X1 to X3 is an electron donating group, and wherein at least one of X1 to X3 is an electron withdrawing group.
47. The compound of any of claims 24-27, wherein at least one of X1 to X17 is C-|-C6 alkyl, - OR46, -SR47, -NR48R49, or -NR50COR51 ; and wherein at least one of X1 to X17 is -CN, - C02R40, -S02OR43, -CONR52R53, -COR54, -N02, -SOR41, -S02R55, -POsR^R45, halo, C C6 acyl, trihalomethyl, or -S02NR56R57.
48. The compound of any of claims 24-27, wherein at least one of X1 to X17 is -OR46, -SR47, -NR48R49, or -NR50COR51.
49. The compound of any of claims 24-27, wherein at least one of X1 to X17 is -NR48R49, or - NR50COR51.
50. The compound of any of claims 24-27, wherein at least one of X1 to X17 is -CN, - COzR^ -COR54, -N02, -S02R55, or -S02NR56R57.
51 . The compound of any of claims 24-27, wherein at least one of X1 to X17 is -C02R4°, - COR54, -S02NR56R57 or -S02R55.
52. The compound of any of claims 24-27, wherein at least one of X1 to X17 is -NR48R49, and wherein at least one of X1 to X17 is -CO2R40,-COR54, -S02NR56R57 or -S02R55.
53. The compound of any of claims 24-27, wherein at least one of X1 to X17 is an electron donating group, and wherein at least one of X1 to X17 is an electron withdrawing group.
54. The compound of any of claims 29-37, wherein at least one of X1 to X8 is CrC6 alkyl, - OR46, -SR47, -NR48R49, or -NR50COR51 ; and wherein at least one of X1 to X8 is -CN, - C02R40, -S02OR43, -CONR52R53, -COR54, -N02, -SOR41, -S02R55, -POsR^R45, halo, Ci-C6 acyl, trihalomethyl, or -S02NR56R57.
55. The compound of any of claims 29-37, wherein at least one of X1 to X8 is -OR46, -SR47, -NR48R49, or -NR50COR51.
56. The compound of any of claims 29-37, wherein at least one of X1 to X8 is -NR48R49, or - NR50COR51.
57. The compound of any of claims 29-37, wherein at least one of X1 to X8 is -CN, - COzR^ -COR54, -N02, -S02R55, or -S02NR56R57.
58. The compound of any of claims 29-37, wherein at least one of X1 to X8 is -C02R40, - COR54, -S02NR56R57 or -S02R55.
59. The compound of any of claims 29-37, wherein at least one of X1 to X8 is -NR48R49, and wherein at least one of X1 to X17 is -CO2R40,-COR54, -S02NR56R57 or -S02R55.
60. The compound of any of claims 29-37, wherein at least one of X1 to X8 is an electron donating group, and wherein at least one of X1 to X8 is an electron withdrawing group.
61. The compound of any of claims 1-60, wherein each of R1 and R5 is hydrogen.
62. The compound of any of claims 1-61 , wherein each of R1, R2,R4, and R5 is hydrogen.
63. The compound of any of claims 1-60, wherein R3 is -OCF3.
64. The compound of any of claims 1-7, 10-16, 20-21 , 23-37, wherein R3 is PS1 or PS2.
65. The compound of any of claims 12-18, 20, and 23-37, wherein at least one of X1 to X17 is PS1 or PS2.
66. The compound of any of claims 1 - 65, wherein the compound inhibits the biological activity of a matrix metalloproteinase enzyme.
67. The compound of claim 66, wherein the matrix metalloproteinase enzyme is Matrix
Metalloproteinase-2, Matrix Metalloproteinase-9, or Matrix Metalloproteinase-13.
68. The compound of claim 66, wherein the compound selectively binds to a matrix
metalloproteinase receptor of a target tissue.
69. A compound being of the formula (FX105), (FX106), (FX107) or (FX108):
Figure imgf000158_0001
(FX105);
Figure imgf000159_0001
70. The compound of any of claims 1-69 for use in an optical imaging, diagnostic, and/or therapeutic biomedical procedure.
71. The compound of claim 70, wherein the biomedical procedure comprises: administering to a subject a therapeutically or diagnostically effective amount of the compound under conditions sufficient for contacting a target tissue or target cell of the subject with the compound of any of claims 1-69, wherein the compound selectively binds to a matrix metalloproteinase enzyme expressed by the target tissue or target cell; and exposing the administered compound to a therapeutically or diagnostically effective amount of electromagnetic radiation.
72. The compound of claim 71 , wherein the procedure comprises exposing the administered compound to electromagnetic radiation having wavelengths selected over the range of 350 nanometers to 1300 nanometers.
73. The compound of any of claims 71 -72, wherein the procedure comprises detecting
electromagnetic radiation emitted from the compound administered to the subject.
74. The compound of any of claims 71 -73, wherein exposing the compound administered to the subject to electromagnetic radiation generates a diagnostically effective amount of fluorescence from the compound.
75. The compound of claim 74, wherein the fluorescence from the compound has
wavelengths selected over the range of 500 nanometers to 1300 nanometers.
76. The compound of any of claims 74-75, wherein the procedure comprises generating an image of the fluorescence from the compound.
77. The compound of claim 71 , wherein the procedure is a Type 1 or Type 2 phototherapy procedure.
78. The compound of claim 77, wherein exposing the administered compound to
electromagnetic radiation generates a therapeutically effective amount of reactive species causing localized cell death or injury.
79. The compound of any of claims 71 -78, wherein the procedure comprises accumulating the compound at the target cell of a selected organ in the subject.
80. The compound of any of claims 71 -79, wherein the procedure comprises accumulating the compound at the target cell of a selected tissue type in the subject.
81. The compound of claim 80, wherein the tissue type is colon, prostate, gastric,
esophageal, uterine, endometrial, pancreatic, breast, cervical, brain, skin, gallbladder, lung, or ovary.
82. The compound of claim 80, wherein the tissue type is cancerous tissue.
83. The compound of claim 80, wherein the tissue type is a tumor.
84. The compound of any of claims 1-83 for use in diagnosis or treatment of inflammation or an inflammation-associated disorder.
85. The compound of claim 84, wherein the inflammation or inflammation-associated
disorder is osteoarthritis or rheumatoid arthritis.
86. The compound of any of claims 1-83 for use in diagnosis or treatment of cancer or a cancer-associated disorder.
87. The compound of claim 86, wherein the cancer or cancer-associated disorder is colon cancer, prostate cancer, gastric cancer, esophageal cancer, uterine cancer, endometrial cancer, pancreatic cancer, breast cancer, cervical cancer, brain cancer, skin cancer, gallbladder cancer, lung cancer, or ovarian cancer
88. A method of detecting a cell or tissue expressing a matrix metalloproteinase enzyme comprising: administering to the cell or tissue expressing the matrix metalloproteinase enzyme a therapeutically or diagnostically effective amount of a compound of any of claims 1 - 69 under conditions sufficient for contacting the cell or tissue expressing matrix
metalloproteinase enzyme with the compound; exposing the administered compound to a diagnostically effective amount of
electromagnetic radiation having wavelengths selected over the range of 350 nm and 1300 nm; and detecting electromagnetic radiation emitted from the compound.
89. A method for carrying out a phototherapy procedure comprising: administering to a subject a therapeutically effective amount of a compound of any of claims 1 - 69 under conditions sufficient for contacting a target tissue of the subject with the compound, wherein the compound selectively binds to a matrix metalloproteinase enzyme expressed by the target tissue; and exposing the administered compound to a therapeutically effective amount of electromagnetic radiation having wavelengths selected over the range of 350 nm and 1300 nm, thereby resulting in injury or death to cells at the target tissue.
90. A pharmaceutical composition comprising a compound of any of claims 1 - 89 and one or more pharmaceutically acceptable excipients.
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CN104119856B (en) * 2013-04-26 2016-08-17 中国科学院大连化学物理研究所 The one hypochlorous fluorescent probe of class ratio test and the application in living things system thereof
WO2017132642A1 (en) * 2016-01-29 2017-08-03 The Regents Of The University Of California A wearable sensor, and method, to monitor anti-coagulation therapy
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US10988466B2 (en) 2017-03-23 2021-04-27 Jacobio Pharmaceuticals Co., Ltd. Heterocyclic derivatives useful as SHP2 inhibitors

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