Composition of near IR closed chain, thiocyanine dye and prostate specific membrane antigen ligand

Abstract

The present invention is a composition of a near-IR closed chain, thiocyanine dye and a prostate-specific membrane antigen ligand. A composition and method for visualizing tissue under the irradiation of near-infrared radiation using a fusion compound of a near-IR, closed chain, thiocyanine dye and a prostate-specific membrane antigen ligand are provided.

Description

[0001] This application is a divisional application. The application date of the original application is April 11, 2017, the application number is 201780017832.6, and the name of the invention is "Compositions of near-IR closed-chain, thiocyanine dyes and prostate-specific membrane antigen ligands".

[0002] Cross-references to related application data

[0003] This application claims the benefit of U.S. Nonprovisional Patent Application No. 15 / 261,357, filed on September 9, 2016, entitled “COMPOSITIONS OF NEAR IR CLOSED CHAIN, SULFO-CYANINE DYES AND PROSTATE SPECIFIC MEMBRANE ANTIGEN LIGANDS,” and Provisional Application No. 62 / 324,097, filed on April 18, 2016, entitled “COMPOSITIONS OF NEAR IR CLOSED CHAIN, SULFO-CYANINE DYES AND PROSTATE SPECIFIC MEMBRANE ANTIGEN LIGANDS,” which are incorporated herein by reference in their entirety for all purposes. Background Art

[0004] Minimally invasive medical technology is intended to reduce the amount of irrelevant tissue damaged during the diagnosis or surgical procedure, thereby reducing patient recovery time, discomfort and harmful side effects. Although millions of "open" or traditional surgical procedures are performed in the United States every year, many of these surgical procedures can potentially be performed in a minimally invasive manner. For example, one effect of minimally invasive surgery is the reduced postoperative recovery time and the associated hospital stay. Because the average hospital stay for standard surgical procedures is usually significantly longer than the average stay for similar minimally invasive surgical procedures, the use of minimally invasive technology increases and can save millions of dollars in hospital costs each year. Although many of the surgical procedures performed in the United States can potentially be performed in a minimally invasive manner, due to instrument limitations, method limitations and the additional surgical training involved in mastering the technology, only a portion of these technologies are currently used.

[0005] Minimally invasive telesurgery systems are being developed to increase the proficiency of surgeons and to allow surgeons to operate on patients from a remote location. Telesurgery is a general concept of a surgical system in which the surgeon uses some form of remote control (e.g., servomechanisms, etc.) to manipulate the movement of surgical instruments rather than directly holding and moving the instruments by hand. In such telesurgery systems, an image of the surgical site is provided to the surgeon at a remote location. While viewing the surgical site on a suitable viewer or screen, the surgeon performs a surgical procedure on the patient by manipulating the main control input device, which in turn controls the movement of the instrument. These input devices can move the working end of the surgical instrument sufficiently proficiently to perform very complex surgical tasks.

[0006] Minimally invasive medical techniques, including telesurgery systems, can be further aided by improving visualization of tissue to be operated on. One way to improve visualization of tissue is by using dyes that can target visualized tissue, allowing the surgeon to remove or spare the tissue. Therefore, there is a need for new fusion compositions of near infrared (IR), closed chain, thio-cyanine dyes and prostate specific membrane antigen (PSMA) ligands and new methods for visualizing tissue under illumination with near infrared radiation. Surprisingly, the present invention meets these and other needs. Summary of the invention

[0007] The present invention generally provides novel compositions and methods for visualizing tissue under irradiation using fusion compounds, including near infrared radiation using near IR, closed chain, thio-cyanine dyes and prostate specific membrane antigen ligands.

[0008] In a first embodiment, the present invention provides a composition comprising a compound having the formula:

[0009]

[0010] in,

[0011] R 1 , R 2 , R 3 , R 4 , R 5 , R 6 and R 7 Each is independently hydrogen, C 1-4 Alkyl, substituted C 1-4 Alkyl or -CO2T;

[0012] X is a single bond, -O- or -S-;

[0013] The subscripts a, b, c, d, e, f, g, and h are each independently an integer from 1 to 6, and T is each independently a metal ion, H, or a negative charge;

[0014] Z is each independently H, C 1-6 Substituted or unsubstituted alkyl, C 6-12 Substituted or unsubstituted aryl, or C 6-12 substituted or unsubstituted heteroaryl; and

[0015] Among other things, the composition is suitable for visualizing tissue under irradiation with near infrared radiation.

[0016] In some cases, the composition has the formula:

[0017]

[0018] In some cases, the composition has the formula:

[0019]

[0020] In some cases, the composition is a pharmaceutical composition in a unit dosage form comprising a compound of Formula I, II or III, wherein the composition is suitable for administration to a patient; and wherein the unit dosage form of the compound is delivered to the patient in an amount between 0.01 and 8 mg / kg. In some cases, the composition unit dosage form is delivered to the patient in an amount of 0.01, 0.05, 0.10, 0.20, 0.30, 0.35, 0.40, 0.45, 0.50, 0.55, 0.60, 0.65, 0.70, 0.75, 0.80, 0.90, 1, 2, 4, 6 or 8 mg / kg. In some cases, the composition is dry and in single dose form.

[0021] In some embodiments, the composition is lyophilized in a sterile container. In some cases, the composition is contained in a sterile container, wherein the container has a machine-detectable identifier that can be read by a medical device.

[0022] In some embodiments, the composition further comprises a combination with a pharmaceutically acceptable excipient in an oral dosage form. In some embodiments, the composition further comprises a combination with a pharmaceutically acceptable carrier in an injectable dosage form. In some embodiments, the composition further comprises a combination with a pharmaceutically acceptable excipient in a dosage form for direct delivery to a surgical site.

[0023] In some embodiments, the present invention provides a use of a composition suitable for administration to a patient to obtain visualization of PSMA-expressing tissue under irradiation with near-infrared radiation, wherein the unit dosage form is delivered to the patient in an amount between 0.01 and 8 mg / kg. In some cases, the use is suitable for administration to a human patient to obtain visualization of human tissue under irradiation with near-infrared radiation, wherein the unit dosage form is delivered to the patient in an amount between 0.01 and 8 mg / kg.

[0024] In some embodiments, the present invention provides a method for visualizing tissue expressing PSMA, the method comprising administering to a patient a composition of a compound of Formula I, II or III described herein. In some embodiments, the method comprises administering to a patient a composition of a compound having the formula:

[0025]

[0026] in,

[0027] R 1 , R 2 , R 3 , R 4 , R 5 , R 6 and R 7 Each is independently hydrogen, C 1-4 Alkyl, substituted C 1-4 Alkyl or -CO2T;

[0028] X is a single bond, -O- or -S-;

[0029] The subscripts a, b, c, d, e, f, g, and h are each independently an integer from 1 to 6;

[0030] Each T is independently a metal ion, H or a negative charge; each Z is independently H, C 1-6 Substituted or unsubstituted alkyl, C 6-12 Substituted or unsubstituted aryl, or C 6-12 substituted or unsubstituted heteroaryl;

[0031] and wherein the compound is administered in an amount sufficient to image the tissue upon exposure to near infrared radiation;

[0032] imaging the tissue under illumination with near infrared radiation; and obtaining at least one image of the tissue from the patient using the composition.

[0033] In some embodiments, the method administers a pharmaceutical composition to a patient, the pharmaceutical composition comprising a unit dosage form of a compound of Formula I, wherein the composition is sterile, nontoxic and suitable for administration to a patient; and wherein the unit dosage form of the compound is delivered to the patient in an amount between 0.01 and 8 mg / kg. In some cases, the method further comprises obtaining an image during administration of the composition, after administration of the composition, or during and after administration of the composition. In some cases, the method further comprises intravenously injecting a composition of Formula I into a patient. In some cases, the composition is injected into the circulatory system.

[0034] In some embodiments, the method further includes visualizing a patient area on which a surgical procedure is performed or to be performed, or visualizing a patient area in order to observe a patient area otherwise examined by a medical professional. In some cases, the method further includes performing a surgical procedure on the patient area based on the visualization of the surgical area. In some cases, the method further includes observing a patient area on which an eye, arthroscopic, laparoscopic, cardiothoracic, muscle, or neurological procedure is performed or to be performed. In some cases, the method further includes obtaining an ex vivo image of at least a portion of the patient.

[0035] In some embodiments, the visualized tissue is tumor tissue. In some cases, the visualized tissue is dysplastic or cancerous tissue. In some cases, the visualized tissue is prostate tissue. In some cases, the visualized tissue is prostate tumor tissue. BRIEF DESCRIPTION OF THE DRAWINGS

[0036] FIG1 shows four fluorescence images. Each of the four fluorescence images shows similar tumor tissue treated with a different fusion compound including a different dye and a targeting ligand. The treated tumor tissue in each of the four images was exposed to narrow-band near-IR excitation light. The narrow-band near-IR excitation light includes a wavelength of light expected to correspond to the excitation maximum wavelength of each fluorophore associated with each of the fusion compounds.

[0037] ο Figure 1A Tumor tissue treated with a fusion compound having the composition DyLight800+PSMA targeting ligand is shown.

[0038] ο Figure 1B Tumor tissue treated with a fusion compound having the composition ZW800 + PSMA targeting ligand is shown. ZW800 is ZW800-1 manufactured by FLARE Foundation or Curadel.

[0039] ο Figure 1CTumor tissue treated with a fusion compound having the composition IRDye800CW + PSMA targeting ligand is shown. IRDye800CW is manufactured by Li-Cor.

[0040] ο Figure 1D Tumor tissue treated with a fusion compound having a targeting ligand consisting of Indocyanine green (ICG) + PSMA is shown.

[0041] Figure 2 A histogram depicting the average signal intensity detected by the imaging sensor when each image was taken is shown. The "I" shaped bar at the top of each histogram corresponds to the standard deviation of the signal intensity across the array of pixels that make up the image sensor.

[0042] o Columns labeled Dylight correspond to DyLight800CW+PSMA targeting ligand.

[0043] o Columns labeled IRDye800 correspond to IRDye800+PSMA targeting ligand.

[0044] o The column labeled ZW800 corresponds to ZW800+PSMA targeting ligand.

[0045] o Columns labeled with ICG correspond to ICG+PSMA targeting ligand.

[0046] o The column labeled "control" corresponds to tumor tissue that was not treated with any type of fluorophore-containing fusion compound. Due to the inherent noise of the fluorescence imaging device used, the average signal intensity corresponding to the control is higher than the average signal intensity corresponding to ZW800 and ICG. DETAILED DESCRIPTION

[0047] I. Overview

[0048] The present invention provides novel compositions and methods for visualizing tissue under illumination with near-infrared radiation using fusion compounds comprising near-IR, closed-chain, thio-cyanine dyes and prostate-specific membrane antigen (PSMA) ligands. Surprisingly, the fusion compounds of the present invention show significantly increased fluorescence compared to other combinations of dyes and PSMA ligands.

[0049] II. Definitions

[0050] Abbreviations used herein have their conventional meanings in the chemical and biological fields.

[0051] As used herein, the term "metal ion" refers to an element in the periodic table that is metallic and positively charged, thus having fewer electrons in the valence electron shell than in a neutral metal element. Useful metals in the present invention include metals that can form pharmaceutically acceptable compositions. Useful metals include Li, Na, K, Rb, Cs, Be, Mg, Ca, Sr and Ba. Those skilled in the art will recognize that the metals described above can each adopt several different oxidation states. In some examples, the most stable oxidation state is formed, but other oxidation states are also useful in the present invention.

[0052] As used herein, the term "alkyl" refers to a straight or branched, saturated, aliphatic free radical with a specified number of carbon atoms. For example, C1-C6 alkyl includes but is not limited to methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, amyl, isopentyl, hexyl, etc. Other alkyl groups include but are not limited to heptyl, octyl, nonyl, decyl, etc. Alkyl can include any number of carbons, such as 1-2, 1-3, 1-4, 1-5, 1-6, 1-7, 1-8, 1-9, 1-10, 2-3, 2-4, 2-5, 2-6, 3-4, 3-5, 3-6, 4-5, 4-6 and 5-6. Alkyl groups are typically monovalent, but can be divalent, such as when alkyl links two parts together.

[0053] As used herein, the term "cycloalkyl" refers to a saturated or partially saturated, unsaturated, monocyclic, fused bicyclic or bridged polycyclic ring system containing 3 to 12 ring atoms or the specified number of atoms. Monocyclic rings include, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and cyclooctyl. Bicyclic and polycyclic rings include, for example, norbornane, decalin and adamantane. For example, C 3-8 Cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclooctyl and norbornane.

[0054] As used herein, the term "haloalkyl" refers to an alkyl group as defined above, in which some or all of the hydrogen atoms are replaced by halogen atoms. Halogen (halo) preferably means chloro or fluoro, but may also be bromo or iodo. For example, haloalkyl includes trifluoromethyl, fluoromethyl, 1,2,3,4,5-pentafluoro-phenyl, etc. The term "perfluoro" defines compounds or free radicals having at least two available hydrogens replaced by fluorine. For example, perfluorophenyl refers to 1,2,3,4,5-pentafluorophenyl, perfluoromethane refers to 1,1,1-trifluoromethyl and perfluoromethoxy refers to 1,1,1-trifluoromethoxy.

[0055] As used herein, the term "halogen" refers to fluorine, chlorine, bromine and iodine.

[0056] As used herein, the term "aryl" refers to a monocyclic or fused bicyclic, tricyclic or larger aromatic ring system containing 6 to 16 ring carbon atoms. For example, aryl can be phenyl, benzyl or naphthyl, preferably phenyl. "Arylene" means a divalent radical derived from an aryl group. The aryl group can be one, two or three radicals selected from alkyl, alkoxy, aryl, hydroxyl, halogen, cyano, amino, amino-alkyl, trifluoromethyl, alkylenedioxy and oxy-C2-C3-alkylene, mono-, di- or tri-substituted; all of which are optionally further substituted, such as defined above; or 1- or 2-naphthyl; or 1- or 2-phenanthrenyl substituted. Alkylenedioxy is a divalent substituent attached to two adjacent carbon atoms of phenyl, such as methylenedioxy or ethylenedioxy. Oxy-C2-C3-alkylene is also a divalent substituent attached to two adjacent carbon atoms of phenyl, such as oxyethylene or oxypropylene. An example of oxy-C2-C3-alkylene-phenyl is 2,3-dihydrobenzofuran-5-yl.

[0057] As used herein, the term "heteroaryl" refers to a monocyclic or fused bicyclic or tricyclic aromatic ring system containing 5 to 16 ring atoms, wherein 1 to 4 of the ring atoms are heteroatoms, each of which is N, O or S. For example, heteroaryl includes pyridyl, indolyl, indazolyl, quinoxalinyl, quinolyl, isoquinolyl, benzothiophenyl, benzofuranyl, furanyl, pyrrolyl, thiazolyl, benzothiazolyl, Azolyl, iso Oxylidene, triazolyl, tetrazolyl, pyrazolyl, imidazolyl, thienyl, or any other free radical substituted, especially mono- or di-substituted, such as alkyl, nitro or halogen. Pyridyl represents 2-, 3- or 4-pyridyl, advantageously 2- or 3-pyridyl. Thienyl represents 2- or 3-thienyl. Quinolyl preferably represents 2-, 3- or 4-quinolyl. Isoquinolyl preferably represents 1-, 3- or 4-isoquinolyl. Benzopyranyl, benzothiopyranyl preferably represent 3-benzopyranyl or 3-benzothiopyranyl, respectively. Thiazolyl preferably represents 2- or 4-thiazolyl, and most preferably, 4-thiazolyl. Triazolyl is preferably 1-, 2- or 5-(1,2,4-triazolyl). Tetrazolyl is preferably 5-tetrazolyl.

[0058] Preferably, heteroaryl is pyridyl, indolyl, quinolyl, pyrrolyl, thiazolyl, isothioyl, oxazolyl, triazolyl, tetrazolyl, pyrazolyl, imidazolyl, thienyl, furyl, benzothiazolyl, benzofuranyl, isoquinolyl, benzothiophenyl, oxazolyl, indazolyl, or an optionally substituted, especially mono- or di-substituted, radical.

[0059] Similarly, substituents for aryl and heteroaryl groups are varied and are selected from: -halogen, -OR', -OC(O)R', -NR'R", -SR', -R', -CN, -NO2, -CO2R', -CONR'R", -C(O)R', -OC(O)NR'R", -NR"C(O)R', -NR"C(O)2R', -NR'-C(O)NR"R"', -NH-C(NH2)=NH, -NR'C(NH2)=NH, -NH-C(NH2)= NR', -S(O)R', -S(O)2R', -S(O)2NR'R", -N3, -CH(Ph)2, perfluoro(C1-C4)alkoxy, and perfluoro(C1-C4)alkyl, in a number ranging from zero to the total number of open valences on the aromatic ring system; and wherein R', R" and R'" are independently selected from hydrogen, (C1-C8)alkyl and heteroalkyl, unsubstituted aryl and heteroaryl, (unsubstituted aryl)-(C1-C4)alkyl, and (unsubstituted aryl)oxy-(C1-C4)alkyl.

[0060] As used herein, the term "visualization" refers to a process of obtaining a graphic image of a tissue by any means, including illumination with near infrared radiation.

[0061] The term "near infrared radiation" or "near IR radiation" refers to optical radiation having a wavelength in the range of approximately 700 nm to 1400 nm. Reference herein to the optionally plural term "wavelength(s)" indicates that the radiation may be a single wavelength or a spectrum of radiation having different wavelengths.

[0062] As used herein, the term "patient" refers to mammals, such as rodents, felines, canines, and primates; most preferably the patient is a human.

[0063] As used herein, the term "tissue" includes, but is not limited to, allogeneic or xenogeneic bone, neural tissue, fibrous connective tissue (including tendons and ligaments), cartilage, dura mater, fascia, pericardium, muscle, heart valves, veins and arteries and other blood vessels, dermis, adipose tissue, glandular tissue, prostate tissue, kidney tissue, brain tissue, renal tissue, bladder tissue, lung tissue, breast tissue, pancreatic tissue, vascular tissue, tumor tissue, cancerous tissue, or prostate tumor tissue.

[0064] As used herein, the term "sterile" refers to a system or components of a system that are free of infectious agents, including bacteria, viruses, and biologically active RNA or DNA.

[0065] As used herein, the term "non-toxic" means that no harmful effects occur when administered to a vertebrate as a result of using the pharmaceutical composition at a level effective for visualizing tissue under irradiation of near infrared radiation (therapeutic level).

[0066] As used herein, the term "unit dosage form" encompasses any amount that can be suitably used to measure the administration of a pharmaceutical composition to a patient. Preferably, the unit dosage form is between 0.01 and 8 mg / kg, or 0.01 and 5 mg / kg, or between 0.01 and 1 mg / kg. Suitable dosage ranges also include 0.01, 0.05, 0.10, 0.20, 0.30, 0.35, 0.40, 0.45, 0.50, 0.55, 0.60, 0.65, 0.70, 0.75, 0.80, 0.90, 1, 2, 4, 6 or 8 mg / kg. As recognized by those skilled in the art herein, when another form (e.g., another salt of a pharmaceutical composition) is used in a preparation, weight can be adjusted to provide an equivalent amount of the pharmaceutical composition.

[0067] As used herein, the term "oral dosage form" refers to its ordinary meaning in the art (ie, a pharmaceutical composition in the form of a tablet, capsule, caplet, gelcap, geltab, pill, etc.).

[0068] As used herein, the term "injectable dosage form" refers to its ordinary meaning in the art (ie, to pharmaceutical compositions in the form of solutions, suspensions, and emulsions, for example, water or water / propylene glycol solutions).

[0069] "Pharmaceutically acceptable excipients" and "pharmaceutically acceptable carriers" refer to substances that help administer active agents to patients and are absorbed by patients and can be included in the compositions of the present invention without causing significant adverse toxicological effects on patients. Non-limiting examples of pharmaceutically acceptable excipients include water, NaCl, physiological saline solution, sodium lactate Ringer's solution, normal sucrose, normal glucose, binders, fillers, disintegrants, lubricants, coatings, sweeteners, spices and pigments, etc. Those skilled in the art will recognize that other pharmaceutical excipients are useful in the present invention. Pharmaceutically acceptable carriers include, but are not limited to, any adjuvant, excipient, glidant, sweetener, diluent, preservative, dye / colorant, flavoring agent, surfactant, wetting agent, dispersant, suspending agent, stabilizer, isotonic agent, solvent or emulsifier.

[0070] As used herein, the term "machine detectable identifier" includes identifiers that can be seen or detected by a machine (including a medical device). In some examples, the medical device is a telesurgery system. Machine detectable identifiers can facilitate the access or utilization of information directly encoded in a machine detectable identifier or stored elsewhere. Examples of machine detectable identifiers include microchips, radio frequency identification (RFID) tags, bar codes (e.g., 1-dimensional or 2-dimensional bar codes), data matrices, quick response (QR) codes, and holograms. Those skilled in the art will recognize that other machine detectable identifiers are useful in the present invention.

[0071] III. Composition

[0072] A. Compounds

[0073] The compounds of the present invention include fusion compounds of Formula I, II and III that absorb light at wavelengths in the near infrared region of the electromagnetic spectrum. In some embodiments, the present invention provides a composition comprising a compound having the formula:

[0074]

[0075] in,

[0076] R 1 , R 2 , R 3 , R 4 , R 5 , R 6 and R 7 Each is independently hydrogen, C 1-4 Alkyl, substituted C 1-4 Alkyl or -CO2T;

[0077] X is a single bond, -O- or -S-;

[0078] The subscripts a, b, c, d, e, f, g, and h are each independently an integer from 1 to 6, and

[0079] Each T is independently a metal ion, H or a negative charge; each Z is independently H, C 1-6 Substituted or unsubstituted alkyl, C 6-12 Substituted or unsubstituted aryl, or C 6-12 substituted or unsubstituted heteroaryl; and

[0080] Among other things, the composition is suitable for visualizing tissue under irradiation with near infrared radiation.

[0081] In some other embodiments, the composition has the formula:

[0082]

[0083] In some other embodiments, the composition has the formula:

[0084]

[0085] The compounds of the present invention may exist as salts. The present invention includes such salts. Examples of applicable salt forms include hydrochlorides, hydrobromides, sulfates, mesylates, nitrates, maleates, acetates, citrates, fumarates, tartrates (such as (+)-tartrates, (-)-tartrates or mixtures thereof including racemic mixtures), succinates, benzoates and salts with amino acids (such as glutamic acid). These salts can be prepared by methods known to those skilled in the art. Also included are base addition salts, such as sodium, potassium, calcium, ammonium, organic amino or magnesium salts, or similar salts. When the compounds of the present invention contain relatively basic functional groups, acid addition salts can be obtained by contacting such compounds in neutral form with sufficient desired acids (either pure or in a suitable inert solvent). Examples of acceptable acid addition salts include those derived from inorganic acids such as hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, monohydrogencarbonic acid, phosphoric acid, monohydrogenphosphoric acid, dihydrogenphosphoric acid, sulfuric acid, monohydrogensulfuric acid, hydroiodic acid or phosphorous acid, etc., and salts derived from organic acids such as acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, tartaric acid, methanesulfonic acid, etc. Also included are salts of amino acids such as arginine, and salts of organic acids such as glucuronic acid or galacturonic acid, etc. Certain specific compounds of the present invention contain both basic and acidic functional groups, which allow the compounds to be converted into base or acid addition salts.

[0086] Other salts include acid or base salts of the compounds used in the methods of the present invention. Illustrative examples of pharmaceutically acceptable salts are inorganic acid (hydrochloric acid, hydrobromic acid, phosphoric acid, etc.) salts, organic acid (acetic acid, propionic acid, glutamic acid and citric acid, etc.) salts and quaternary ammonium (iodomethane, iodoethane, etc.) salts. It should be understood that pharmaceutically acceptable salts are non-toxic. Additional information on suitable pharmaceutically acceptable salts can be found in Remington's Pharmaceutical Sciences, 17th ed., Mack Publishing Company, Easton, Pa., 1985, which is incorporated herein by reference.

[0087] Pharmaceutically acceptable salts include salts of active compounds prepared using relatively nontoxic acids or bases, depending on the specific substituents found on the compounds described herein. When the compounds of the present invention contain relatively acidic functional groups, base addition salts can be obtained by contacting such compounds in a neutral form with a sufficient amount of the desired base (either neat or in a suitable inert solvent). Examples of pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amino or magnesium salts, or similar salts. When the compounds of the present invention contain relatively basic functional groups, acid addition salts can be obtained by contacting such compounds in a neutral form with a sufficient amount of the desired acid (either neat or in a suitable inert solvent). Examples of pharmaceutically acceptable acid addition salts include those derived from inorganic acids and salts derived from relatively nontoxic organic acids, such as hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, monohydrogencarbonic acid, phosphoric acid, monohydrogenphosphoric acid, dihydrogenphosphoric acid, sulfuric acid, monohydrogensulfuric acid, hydroiodic acid or phosphorous acid, etc., organic acids such as acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, tartaric acid, methanesulfonic acid, etc. Also included are salts of amino acids such as arginine, etc., and salts of organic acids such as glucuronic acid or galacturonic acid, etc. (see, e.g., Berge et al., "Pharmaceutical Salts", Journal of Pharmaceutical Science, 1977, 66, 1-19). Certain specific compounds of the present invention contain both basic and acidic functional groups, which allow the compounds to be converted into base or acid addition salts.

[0088] The neutral forms of the compounds are preferably regenerated by contacting the salt with a base or acid and isolating the parent compound in the conventional manner.The parent form of the compound differs from the various salt forms in certain physical properties, such as solubility in polar solvents.

[0089] Some compounds of the present invention may exist in non-solvated form and solvated form, including hydrated form. Generally, solvated form is equivalent to non-solvated form, and is encompassed within the scope of the present invention. Some compounds of the present invention may exist in polycrystalline or amorphous form. Generally, all physical forms are equivalent and are intended to be within the scope of the present invention for the purposes contemplated by the present invention.

[0090] In some embodiments, the present invention provides a pharmaceutical composition comprising a pharmaceutically acceptable excipient.

[0091] B. Synthesis

[0092] The compounds described in the above embodiments can be made using processes known in the art. Generally, fusion compounds of the present invention can be synthesized by attaching nearly IR, closed-chain, thio-cyanine dyes to prostate-specific membrane antigen ligands via bonds. The materials used can be determined by the type of desired structure and bond used.

[0093] For example, the prostate-specific membrane antigen ligand used in the composition of the present invention can be synthesized as described in PCT application WO2010 / 108125, and is incorporated herein in its entirety. Compounds can be assembled by reaction between different components to form bonds, such as urea (-NRC (O) NR-), thiourea (-NRC (S) NR-), amide (-C (O) NR- or -NRC (O)-), or ester (-C (O) O- or -OC (O)-). Urea bonds can be easily prepared by reaction between amine and isocyanate, or between amine and activated carbonylamide (-NRC (O)-). Thiourea can be easily prepared from the reaction of amine and isothiocyanate. Amides (-C (O) NR- or -NRC (O)-), such as acyl halides or N-hydroxysuccinimide esters, can be easily prepared by reaction between amine and activated carboxylic acid or ester. Carboxylic acids can also be activated in situ, for example, using coupling agents, such as carbodiimide, or carbonyl diimidazole (CDI). Esters can be formed by reaction between alcohols and activated carboxylic acids.Triazoles are readily prepared by reaction between azides and alkynes, optionally in the presence of a copper (Cu) catalyst.

[0094] The prostate specific membrane antigen ligand can also be prepared by sequential addition of the components to a preformed urea such as the lysine-urea-glutamate compound described in Banerjee et al. (J. Med. Chern. vol. 51, pp. 4504-4517, 2008). Other urea-based compounds can also be used as building blocks.

[0095] Exemplary syntheses of near IR, closed chain, thio-cyanine dyes used in the compositions of the present invention are described in U.S. Pat. No. 6,887,854 and U.S. Pat. No. 6,159,657, and are incorporated herein in their entirety (Formula 1). In addition, some IR, closed chain, thio-cyanine dyes of the present invention are commercially available, including DyLight 800 (ThermoFisher).

[0096] Formula 1

[0097]

[0098] As mentioned above, the fusion composition of the present invention (Formula 2) can be synthesized via the attachment of a near IR, closed chain, thio-cyanine dye to a prostate specific membrane antigen ligand by reacting an active amine on the ligand with a near IR dye. A variety of near infrared dyes are known in the art that have activated functional groups for reaction with amines.

[0099] Formula 2

[0100]

[0101] C. Preparation

[0102] The compositions of the present invention can be prepared in a variety of oral, parenteral and topical dosage forms. Oral preparations include tablets, pills, powders, sugar-coated tablets, capsules, liquids, lozenges, cachets, gels, syrups, slurries, suspensions, etc. suitable for ingestion by patients. The compositions of the present invention can also be administered by injection, i.e., intravenous, intramuscular, intradermal, subcutaneous, intraduodenal or intraperitoneal. Also, the compositions described herein can be administered by inhalation, for example, intranasally. Additionally, the compositions of the present invention can be administered transdermally. The compositions of the present invention can also be administered intraocularly, by insufflation, by powder, and by aerosol formulations (for examples of steroid inhalers, see Rohatagi, J. Clin. Pharmacol. 35: 1187-1193, 1995; Tjwa, Ann. Allergy Asthma Immunol. 75: 107-111, 1995). Therefore, the present invention also provides a pharmaceutical composition comprising a pharmaceutically acceptable carrier or excipient.

[0103] For preparing the pharmaceutical composition of the present invention, pharmaceutically acceptable carriers can be solid or liquid. Solid form products include powders, tablets, pills, capsules, cachets, suppositories and dispersed particles. Solid carriers can be one or more substances, which can also act as diluents, flavoring agents, binders, preservatives, tablet disintegrants or encapsulating materials. The details of the techniques of preparation and administration are well described in the scientific and patent literature, see, e.g., the latest edition of Remington's Pharmaceutical Sciences, Maack Publishing Co, Easton PA ("Remington's").

[0104] In powders, the carrier is a finely divided solid which is in a mixture with the finely divided active ingredient. In tablets, the active ingredient is mixed with a carrier having the necessary binding properties in suitable proportions and compressed into the desired shape and size. Powders and tablets preferably contain from 5% or 10% to 70% of the compound of the invention.

[0105] Suitable solid excipients include, but are not limited to, magnesium carbonate; magnesium stearate; talc; pectin; dextrin; starch; tragacanth; low melting wax; cocoa butter; carbohydrates; sugars, including but not limited to lactose, sucrose, mannitol or sorbitol, starch from corn, wheat, rice, potato or other plants; cellulose such as methylcellulose, hydroxypropyl methylcellulose or sodium carboxymethylcellulose; and gums including gum arabic and tragacanth; and proteins, including but not limited to gelatin and collagen. If desired, disintegrators or solubilizers may be added, such as cross-linked polyvinyl pyrrolidone, agar, alginic acid, or a salt thereof, such as sodium alginate.

[0106] Suitable coatings are provided for sugar-coated tablet cores, such as concentrated sugar solutions, which may also include gum arabic, talcum, polyvinyl pyrrolidone, carbopol gel, polyethylene glycol and / or titanium dioxide, lacquer solutions, and suitable organic solvents or solvent mixtures. Dyes or pigments may be added to tablets or sugar-coated tablet coatings to carry out product identification or characterize the amount (i.e., dosage) of the active compound. Pharmaceutical compositions of the present invention may also be used, for example, push-fit capsules made of gelatin, and soft sealed capsules made of gelatin and coatings such as glycerol or sorbitol for oral use. Push-fit capsules may include compositions of the present invention mixed with fillers or binding agents such as lactose or starch, lubricants such as talcum or magnesium stearate, and optionally stabilizers. In soft capsules, compositions of the present invention may be dissolved or suspended in suitable liquids, such as fatty oils, liquid paraffin or liquid polyethylene glycol, with or without stabilizers.

[0107] Liquid form preparations include solutions, suspensions, and emulsions, for example, water or water / propylene glycol solutions. For parenteral injection, liquid preparations can be formulated as solutions in aqueous polyethylene glycol solution.

[0108] Aqueous solutions suitable for oral use can be prepared by dissolving the composition of the present invention in water and adding suitable colorants, flavors, stabilizers and thickeners as needed. Aqueous suspensions suitable for oral use can be prepared by dispersing the subdivided active ingredients in water with a viscous material and a dispersant or wetting agent, such as natural or synthetic gums, resins, methylcellulose, sodium carboxymethylcellulose, hydroxypropyl methylcellulose, sodium alginate, polyvinyl pyrrolidone, tragacanth and gum arabic, dispersants or wetting agents such as naturally occurring phosphatides (such as lecithin), condensation products of alkylene oxides with fatty acids (such as polyoxyethylene stearate), condensation products of ethylene oxide with long-chain fatty alcohols (such as heptadecaethyleneoxyhexadecyl alcohol), condensation products of ethylene oxide with partial esters derived from fatty acids and hexitol (such as polyoxyethylene sorbitol monooleate), or condensation products of ethylene oxide with partial esters derived from fatty acids and hexitol anhydrides (such as polyoxyethylene sorbitan monooleate). The aqueous suspension may also contain one or more preservatives such as ethyl or n-propyl p-hydroxybenzoate, one or more coloring agents, one or more flavoring agents and one or more sweetening agents, such as sucrose, aspartame or saccharin. The osmotic pressure of the formulation may be adjusted.

[0109] Also included are solid form preparations that are intended to be converted into liquid form preparations for oral administration shortly before use. These liquid forms include solutions, suspensions, and emulsions. In addition to the active ingredient, these preparations may also contain colorants, flavorings, stabilizers, buffers, artificial and natural sweeteners, dispersants, thickeners, solubilizing agents, and the like.

[0110] Oil suspensions can be prepared by suspending the composition of the present invention in vegetable oil or mineral oil or a mixture of these, vegetable oil such as peanut oil, olive oil, sesame oil or coconut oil, mineral oil such as liquid paraffin. The oil suspension may contain a thickener such as beeswax, hard paraffin or cetyl alcohol. Sweeteners may be added to provide oral products with a good taste, such as glycerol, sorbitol or sucrose. These preparations may be preserved by adding antioxidants such as ascorbic acid. As an example of an injectable oil vehicle, see Minto, J.Pharmacol.Exp.Ther.281:93-102,1997. The pharmaceutical preparation of the present invention may also be in the form of an oil-in-water emulsion. The oil phase may be the vegetable oil or mineral oil described above, or a mixture of these. Suitable emulsifiers include naturally occurring gums such as gum arabic and tragacanth, naturally occurring phospholipids such as soy lecithin, esters or partial esters derived from fatty acids and hexitol anhydrides such as sorbitan monooleate, and condensation products of these partial esters with ethylene oxide such as polyoxyethylene sorbitan monooleate. Emulsions may also contain sweeteners and flavorings, such as in the preparation of syrups and elixirs. Such preparations may also contain laxatives, preservatives, or coloring agents.

[0111] The compositions of the present invention can also be delivered as microspheres for slow release in the body. For example, the microspheres can be formulated for administration via intradermal injection of drug-containing microspheres, which are slowly released subcutaneously (see Rao, J. Biomater Sci. Polym. Ed. 7: 623-645, 1995; as biodegradable and injectable gel formulations (see, e.g., Gao Pharm. Res. 12: 857-863, 1995); or as microspheres for oral administration (see, e.g., Eyles, J. Pharm. Pharmacol. 49: 669-674, 1997). Both transdermal and intradermal routes provide sustained delivery for weeks or months.

[0112] In another embodiment, the composition of the present invention can be formulated for parenteral administration, such as intravenous (IV) administration or administration to the cavity of a body cavity or organ. The formulation administered will generally include a solution of the composition of the present invention dissolved in a pharmaceutically acceptable carrier. Acceptable vehicles and solvents that can be used are water and Ringer's solution (an isotonic sodium chloride). In addition, sterile fixed oils can be conventionally used as solvents or suspension media. For this purpose, any mild fixed oil can be used, including synthetic monoglycerides or diglycerides. In addition, fatty acids such as oleic acid can also be used to prepare injections. These solutions are sterile and generally free of undesirable substances. These preparations can be sterilized by conventional well-known techniques, including radiation, chemical, heating / pressurization and filtration sterilization techniques. The preparation may contain pharmaceutically acceptable auxiliary substances required for approximate physiological conditions, such as pH regulators and buffers, toxicity regulators, such as sodium acetate, sodium chloride, potassium chloride, calcium chloride, sodium lactate, etc. The concentration of the composition of the present invention in these preparations can vary widely, and will be selected based on fluid volume, viscosity, body weight, etc., mainly according to the selected specific mode of administration and the needs of the patient. For IV administration, the preparation can be a sterile injectable preparation, such as a sterile injectable aqueous or oily suspension. The suspension can be prepared according to known techniques using those suitable dispersants or wetting agents and suspending agents. The sterile injectable preparation can also be a sterile injectable solution or suspension in a nontoxic parenterally acceptable diluent or solvent, such as a solution of 1,3-butanediol.

[0113] In another embodiment, the preparation of the composition of the present invention can be delivered by using liposomes, which fuse with the cell membrane or are endocytosed, that is, by using ligands attached to the liposomes, or directly attached to the oligonucleotides, which bind to the surface membrane protein receptors of the cell, resulting in endocytosis. By using liposomes, especially when the liposome surface carries ligands specific to the target cells or otherwise preferentially targets specific organs, it is possible to focus on delivering the composition of the present invention into the target cells in vivo. (See, e.g., Al-Muhammed, J. Microencapsul. 13: 293-306, 1996; Chonn, Curr. Opin. Biotechnol. 6: 698-708, 1995; Ostro, Am. J. Hosp. Pharm. 46: 1576-1587, 1989).

[0114] Lipid-based drug delivery systems include lipid solutions, lipid emulsions, lipid dispersions, self-emulsifying drug delivery systems (SEDDS) and self-microemulsifying drug delivery systems (SMEDDS). In particular, SEDDS and SMEDDS are isotropic mixtures of lipids, surfactants and co-surfactants that can spontaneously disperse in aqueous media and form fine emulsions (SEDDS) or microemulsions (SMEDDS). Lipids that can be used in the formulations of the present invention include any natural or synthetic lipids, including but not limited to sesame seed oil, olive oil, castor oil, peanut oil, fatty acid esters, glycerides, and

[0115] In some embodiments, the compositions of the invention are sterile and generally free of undesirable matter. The compounds and compositions can be sterilized by conventional, well-known techniques, including heat / pressure, gas plasma, steam, radiation, chemical and filtration sterilization techniques.

[0116] For example, terminal heat sterilization can be used to destroy all viable microorganisms in the final formulation. Autoclaves are commonly used to achieve terminal heat sterilization of pharmaceutical products in their final packaging. A typical autoclave cycle in the pharmaceutical industry to achieve terminal sterilization of the final product is at 121°C for 15 minutes. The composition of the present invention can be autoclaved at a temperature of 115 to 130°C for a time period ranging from 5 to 40 minutes with acceptable stability. Preferably, autoclave sterilization is performed at a temperature range of 119 to 122°C for a time period ranging from 10 to 36 minutes.

[0117] The composition can also be sterilized by dry heating, as described in U.S. Pat. No. 6,392,036 by Karlsson et al., which discloses a method for dry heating sterilization that can be used for pharmaceutical preparations. The composition can also be sterilized as described in WO02 / 41925 by Breath Limited, which discloses a rapid method for sterilizing a composition similar to pasteurization. The method requires pumping the composition to be sterilized through stainless steel pipes and rapidly raising the temperature of the composition to about 130-145° C. for about 2-20 seconds, followed by rapid cooling to ambient conditions within a few seconds.

[0118] The composition can also be sterilized by irradiation, as described by Illum and Moeller in Arch. Pharm. Chemi. Sci., Ed. 2, 1974, pp. 167-174. The composition can also be sterilized by UV, x-ray, gamma-ray, electron beam radiation, combustion, baking, and chemical sterilization.

[0119] Alternatively, sterile pharmaceutical compositions according to the present invention can be prepared using antiseptic treatment techniques. Aseptic filling is generally used to prepare pharmaceutical products, which will not be subjected to heat sterilization, but wherein all components are sterile. Sterility is maintained by using sterile materials and a controlled working environment. All containers and devices are preferably sterilized by heat sterilization before filling. Then fill containers (such as, vials, ampoules, infusion bags, bottles or syringes) under aseptic conditions.

[0120] In some embodiments, the compounds and compositions of the present invention are nontoxic and generally have no harmful effects when applied to vertebrates at a level effective for visualizing tissues under the irradiation of near-infrared radiation. The toxicity of the compounds and compositions of the present invention can be assessed by measuring their effects on targets (organisms, organs, tissues or cells). Because individual targets usually have different levels of response to the same dose of compounds, population-level measurements of toxicity are often used, which relate to the probability of the results of a certain individual in a population. The toxicology of the compound can be determined by conventional well-known techniques, including in vitro (outside of a living organism) and in vivo (inside a living organism) studies.

[0121] For example, when evaluating the toxicity of a compound, the determination of metabolic stability is usually checked because it is one of the several main determining factors (determinate) that limit the oral bioavailability and systemic clearance of the compound. After oral administration of the compound, it first encounters metabolic enzymes in the gastrointestinal cavity and the intestinal epithelium. After it is absorbed into the bloodstream through the intestinal epithelium, it is first transported to the liver through the portal vein. The compound can be effectively cleared by intestinal or liver metabolism and then reach the systemic circulation, which is called the process of first-pass metabolism. The stability of the compound to metabolism in the liver and extrahepatic tissues will ultimately determine the concentration of the compound found in the systemic circulation and affect its half-life and residence time in the body. Cytochrome P450 (CYP) enzymes are mainly found in the liver, but are also found in the intestinal wall, lungs, kidneys and other extrahepatic organs, and are the main enzymes involved in the metabolism of compounds. Many compounds are inactivated directly or by promoting excretion in the body by CYP. In addition, many compounds are bioactivated by CYP to form their active compounds. Therefore, determining the reactivity of the compound to the CYP enzyme is usually used to evaluate the metabolic stability of the compound.

[0122] The Ames reverse mutation assay is another commonly used toxicology assay for evaluating the toxicity of compounds. The Ames assay utilizes several different tester strains, each of which has different mutations in one of the genes including the histidine (his) biosynthetic operon (Ames, BN et al., (1975) Mutation Res.31:347-363). The detection of revertant (i.e., mutant) bacteria in the test sample that can grow in the absence of histidine indicates that the compound being evaluated is characterized by genotoxic (i.e., mutagenic) activity. The Ames assay is capable of detecting several different types of mutations (genetic damage) that may occur in one or more test strains. The practice of using in vitro bacterial assays to evaluate the genotoxic activity of candidate drugs is based on the prediction that substances that are mutagenic in bacteria may be carcinogenic in experimental animals, and by extension may be carcinogenic or mutagenic to humans.

[0123] In addition, human ether-a-go-go related gene (hERG) assays can be used to assess the potential cardiotoxicity of compounds. Cardiotoxicity may occur when the QT interval is prolonged, leading to an increased risk of life-threatening arrhythmias. The QT interval is a measure of the time between the start of the Q wave and the end of the T wave in the cardiac electrical cycle. The QT interval represents the electrical depolarization and repolarization of the ventricles. A variable QT interval is most often associated with a loss of current through the hERG potassium ion channel, which is due to direct blocking of the ion channel by drugs or by inhibiting the plasma membrane expression of the channel protein (Su et al., J. Biomol Screen 2011, 16, 101–111). Therefore, in vitro hERG screening assays can be used to detect the disruption or inhibition of hERG membrane transport function and assess the potential cardiotoxicity of compounds.

[0124] Other methods of assessing the toxicity of a compound involve administering a relatively large dose of a test compound to a group of animals to determine a level that is lethal to a percentage of the population (mean lethal dose LD). 50 or mean lethal concentration LC 50 The toxicity of a compound can also be assessed in vivo by examining whether the compound produces statistically significant negative effects on the heart, blood pressure, central nervous system (CNS), body weight, food intake, gross or microscopic pathology, clinical pathology, or respiratory measures in animals.

[0125] For example, in a set of in vitro studies evaluating the metabolic stability of the composition of formula III, it was shown that the compound appeared to be stable in rat, dog and human plasma, did not appear to be decomposed into metabolites, and did not show any significant reactivity to 9 major CYP liver enzymes. Additionally, the composition of formula III did not show any mutagenicity at any dose tested in the Ames reverse mutation assay (a widely used in vitro method that determines the ability of a chemical substance to cause DNA mutations). Further safety pharmacology was evaluated in vitro using the human ether-a-go-go related gene (hERG) assay for determining possible cardiotoxic effects. Studies using this assay determined that the composition of formula III only showed a small inhibition (11%) of hERG function at the highest tested concentration (30 times higher than the level effective for visualization), indicating that even at this concentration, the composition of formula III was unlikely to be converted into any clinically threatening physiological cardiac changes.

[0126] In addition, the toxicology of the composition of formula III is also studied in non-clinical in vivo studies in rats and dogs. The study of the effect of the composition of formula III on the heart and respiratory safety of the dog monitored by radio telemetry shows that there is no statistically significant negative effect on the heart, blood pressure or respiratory measurement under a dosage up to 80 times higher than the dosage required for visualization. In rats, additional safety pharmacology indications exceeding 28 days are carried out, and the dosage / day of up to 160 times of the visualization level shows that there is no physiologically significant effect on the central nervous system (CNS). Finally, a series of toxicological studies in dogs show that the dosage of up to 100 times of the visualization level has no significant effect on body weight, food intake, gross pathology or microscopic pathology and clinical pathology (clinical serum chemistry, hematology, coagulation and urinalysis). In rats, dogs or pig animal models, no mortality is observed under any test dose of the composition of formula III.

[0127] In some embodiments, the compositions of the present invention can be lyophilized in sterile containers for dry storage and transportation. The freeze-dried composition can then be reconstituted with sterile water to make a ready-to-use product. The terms "lyophilization", "freeze-dried", and "freeze drying" refer to the process of first freezing the material to be dried and then removing the ice or freezing solvent by sublimation in a vacuum environment. Excipients can be included in the pre-lyophilized formulation to enhance the stability of the freeze-dried product during storage.

[0128] In some embodiments, the composition can be contained in a sterile container, wherein the container has a machine detectable identifier that can be read by a medical device. Examples of machine detectable identifiers include microchips, radio frequency identification (RFID) tags, bar codes (e.g., 1-dimensional or 2-dimensional bar codes), data matrices, quick response (QR) codes, and holograms. Those skilled in the art will recognize that other machine detectable identifiers are useful in the present invention.

[0129] In some cases, the machine-detectable identifier may include a microchip, an integrated circuit (IC) chip, or an electronic signal from a microchip, which can be detected and / or read by a computer system communicating with the medical device. In some cases, the machine-detectable identifier includes a radio frequency identification (RFID) tag. RFID tags are sometimes referred to as transponders. RFID tags are generally devices formed by IC chips, antennas, and adhesive materials, and are used to send or receive predetermined data using an external reader or interrogator. RFID tags can use a reader to send or receive data using a contactless method. Depending on the amplitude of the frequency used, inductive coupling, backscattering, and surface acoustic waves (SAW) can be used. Using electromagnetic waves, data can be sent to or received from a reader by using a full-duplex method, a half-duplex (HDX) method, or a sequential (SEQ) method.

[0130] In some cases, the machine detectable identifier may include a barcode. Barcodes include any machine readable format, including one-dimensional and two-dimensional formats. One-dimensional formats include, for example, Universal Product Code (UPC) and Reduced Space Symbol (RSS). Two-dimensional formats or machine readable matrices include, for example, Quick Response (QR) codes and Data Matrix.

[0131] In some cases, the medical device can be configured to detect machine detectable identifiers. In one example, the medical device is a telesurgery system, which includes a special imaging mode (e.g., fluorescence imaging mode) used with a dye, such as those described in the present disclosure. An example of a telesurgery system including a fluorescence imaging mode is described in US 8,169,468, which is named "Augmented Stereoscopic Visualization for a Surgical Robot", which is incorporated herein in its entirety. In some cases, the medical device can be incorporated into an imaging device, which can scan, read, observe or otherwise detect machine detectable identifiers displayed to the imaging device. In one aspect, only when the medical device detects the presence of a known machine detectable identifier, the medical device will allow the user to access the fluorescence imaging mode of the medical device, which corresponds to the dye that is identified as compatible with the medical device. On the contrary, if the medical device does not detect a known machine detectable identifier, the medical device will not allow the user to access the fluorescence imaging mode and related functions. Imaging devices can include optical scanners, barcode readers, cameras and imaging devices included in telesurgery systems, such as endoscopes. Then, the imaging device can be used to retrieve information associated with machine-detectable identifiers by medical devices. After the identifier is detected, an automatic process can be started to cause a predetermined action to occur or to retrieve or access certain data. The information encoded on the machine-detectable identifier can include instructions for triggering actions, such as, applying the composition of the present invention to the patient. In some embodiments, the machine-detectable identifier includes unencrypted electronic pedigree information of a desired format. The electronic pedigree information can include, for example, batches, effectiveness, expiration, national drug codes, electronic product codes, manufacturers, distributors, wholesalers, pharmacies and / or unique identifiers of saleable units.

[0132] In some embodiments, a sterile container having a machine-detectable identifier includes a fluid outlet configured to mate with a medical device. In some cases, the fluid outlet of the machine-detectable identifier is mechanically secured to the medical device.

[0133] D. Application

[0134] The compounds and compositions of the invention may be delivered by any suitable means, including oral, parenteral, and topical methods.

[0135] The compounds and compositions of the present invention may be delivered by any suitable means, including oral, parenteral and topical methods. Transdermal administration methods by topical routes may be formulated as applicator sticks, solutions, suspensions, emulsions, gels, creams, ointments, pastes, jellies, coatings, powders and aerosols.

[0136] The pharmaceutical preparation is preferably in unit dosage form. In this form, the preparation is subdivided into unit doses containing appropriate quantities of the compounds and compositions of the present invention. The unit dosage form can be a packaged preparation containing discrete quantities of the preparation, such as packaged tablets, capsules, and powders in vials or ampoules. Also, the unit dosage form can be a capsule, tablet, cachet, or lozenge itself, or it can be any of these in an appropriate number of packaged forms.

[0137] In some embodiments, co-administration can be achieved by co-formulation, ie, preparing a single pharmaceutical composition including the compounds and compositions of the invention and any other agents. Alternatively, the various components can be formulated separately.

[0138] The compositions of the present invention and any other agents may be present in any suitable amount and may depend on various factors, including but not limited to the weight and age of the patient, the condition of the disease, etc. Suitable dosage ranges include from about 0.01 and 8 mg / kg, or about 0.01 and 5 mg / kg, or about 0.01 and 1 mg / kg. Suitable dosage ranges also include 0.01, 0.05, 0.10, 0.20, 0.30, 0.35, 0.40, 0.45, 0.50, 0.55, 0.60, 0.65, 0.70, 0.75, 0.80, 0.90, 1, 2, 4, 6, or 8 mg / kg.

[0139] The compositions may also contain other compatible components. The components described herein may be used in combination with each other, with other active components known to be useful for visualizing tissue under illumination with near infrared radiation, or with components that may not be effective alone but may aid in the efficacy of the active components.

[0140] The terms and expressions used herein are used as terms of description rather than limitation, and it is not intended that these terms and expressions be used to exclude equivalent forms of the features shown and described, or portions thereof, recognizing that various modifications are possible within the scope of the claimed invention. Moreover, any one or more features of any embodiment of the invention may be combined with any one or more other features of any other embodiment of the invention without departing from the scope of the invention. For all purposes, all publications, patents, and patent applications cited herein are incorporated herein by reference in their entirety.

[0141] IV. Methods for Visualizing Organizations

[0142] The present invention generally provides novel compositions and methods for visualizing tissue under illumination with near-infrared radiation using fusion compounds comprising a near-IR, closed-chain, thio-cyanine dye and a prostate-specific membrane antigen ligand.

[0143] In some embodiments, the present invention provides a method for visualizing tissue expressing PSMA, the method comprising administering to a patient a composition of a compound having the formula:

[0144]

[0145] in,

[0146] R 1 , R 2 , R 3 , R 4 , R 5 , R 6 and R 7 Each is independently hydrogen, C 1-4 Alkyl, substituted C 1-4 Alkyl or -CO2T;

[0147] X is a single bond, -O- or -S-;

[0148] The subscripts a, b, c, d, e, f, g and h are each independently an integer from 1 to 6,

[0149] Each T is independently a metal ion, H or a negative charge; each Z is independently H, C 1-6 Substituted or unsubstituted alkyl, C 6-12 Substituted or unsubstituted aryl, or C 6-12 substituted or unsubstituted heteroaryl; and wherein the compound is administered in an amount sufficient to image the tissue upon exposure to near infrared radiation;

[0150] imaging the tissue under illumination with near infrared radiation; and obtaining at least one image of the tissue from the patient using the composition.

[0151] In some embodiments, the method administers a pharmaceutical composition to a patient, the pharmaceutical composition comprising a unit dosage form of a compound of Formula I, wherein the composition is sterile, nontoxic and suitable for administration to a patient; and wherein the unit dosage form of the compound is delivered to the patient in an amount between 0.01 and 8 mg / kg. In some cases, the method further comprises obtaining an image during administration of the composition, after administration of the composition, or during and after administration of the composition. In some cases, the method further comprises intravenously injecting a composition of Formula I into a patient. In some cases, the composition is injected into the circulatory system.

[0152] In some embodiments, the present invention provides a composition suitable for administration to a patient to obtain visualization of PSMA-expressing tissue under irradiation with near-infrared radiation, wherein the unit dosage form is delivered to the patient in an amount between 0.01 and 8 mg / kg. In some cases, the use is suitable for administration to a human patient to obtain visualization of human tissue under irradiation with near-infrared radiation, wherein the unit dosage form is delivered to the patient in an amount between 0.01 and 8 mg / kg.

[0153] In some embodiments, the method further includes visualizing a patient area on which a surgical procedure is performed or to be performed, or visualizing a patient area to facilitate viewing of a patient area otherwise examined by a medical professional. In some cases, the method further includes performing a surgical procedure on the patient area based on the visualization of the surgical area. In some cases, the method further includes viewing a patient area on which an eye, arthroscopic, laparoscopic, cardiothoracic, muscle, or neurological procedure is performed or to be performed. In some cases, the method further includes obtaining an ex vivo image of at least a portion of the patient.

[0154] In some embodiments, improved visualization of tissue can be achieved through the use of dyes capable of targeted visualization of tissue, including through fusion compositions of near IR, closed chain, thio-cyanine dyes, and prostate specific membrane antigen (PSMA) ligands. PSMA, while expressed in the prostate tumor epithelium, is also expressed in the neovascularization of many solid tumors (Chang et al., Cancer Res., vol. 59, pp. 3192-3198, 1999; Chang et al., Clin. Cancer Res., vol. 5, pp. 2674-2681, 1999; Gong et al., Cancer Metastasis Rev., vol. 18, pp. 483-490, 1999; Chang et al., Mol. Urol., vol. 3, pp. 313-320, 1999; Baccala et al., Urology, vol. 70, pp. 385-390, 2007; Chang et al., Urology, vol. 57, pp. 801-805, 2001; Milowsky et al., J. Clin. Oncol., vol. 25, pp. 540-547, 2007). Thus, PSMA ligands have been used to mark kidney, bladder, lung, breast, colorectal and pancreatic tumors (Milowsky et al., J. Clin. Oncol., vol. 25, pp. 540-547, 2007). PSMA ligands have been shown to be effective vascular targeting agents in human subjects. Other reports have further investigated PSMA expression in certain tumor types. For example, Baccala et al. noted that clear cell renal cell carcinoma expresses more PSMA in its new blood vessels than in papillary variants (Baccala et al., Urology, vol. 70, pp. 385-390, 2007). In addition, it has been found that angiomyolipoma, a benign renal lesion, does not express PSMA. As an enzyme with an extracellular active site, PSMA represents an excellent target for imaging and treatment of new blood vessels in solid tumors in addition to prostate cancer itself. PSMA-based agents can report on the presence of this marker, which is increasingly recognized as an important prognostic determinant in PCa (Murphy et al., Urology, vol. 51, pp. 89-97, 1998). It is also the target of a variety of new PCa therapies (Gal sky et al., J Clin Oneal, vol. 26, pp. 214 7-2154, 2008).

[0155] Thus, the compositions of the invention comprising a fusion compound of a near-IR, closed-chain, thio-cyanine dye and a PSMA ligand can be used to visualize a variety of tumor tissues. In some cases, the tumor tissue visualized is prostate tissue, kidney tissue, brain tissue, renal tissue, bladder tissue, lung tissue, breast tissue, pancreatic tissue, vascular tissue, tumor tissue, cancerous tissue, or prostate tumor tissue. In some cases, the visualized tissue is cancerous tissue. In some cases, the visualized tissue is prostate tissue. In some cases, the visualized tissue is prostate tumor tissue.

[0156] In some cases, the method further includes the use of the composition of the present invention in combination with the firefly fluorescence of the da Vinci surgical system, which provides an enlarged view that enhances difficult-to-visualize tissues to increase surgical effectiveness, reduce injury rates, and possibly speed up surgery by providing confidence and security to the surgeon that he / she is removing all the expected tissue. For example, the positive margin rate during prostate cancer surgery can be as high as 30%, which means that despite attempts to remove all cancerous tissue, cancerous tissue remains in the male. These patients frequently require follow-up treatments, including but not limited to external beam radiation radiotherapy, brachytherapy, ablation, and hormone therapy. In addition, erectile dysfunction and urinary incontinence rates caused by nerve damage during prostate surgery may be close to 50% and 20%, respectively. Compositions of the present invention comprising fusion compounds of near-IR, closed-chain, thio-cyanine dyes and PSMA ligands can also be used to increase the contrast between prostate and nerve tissue, helping to avoid damage to nerve and sphincter tissue. Thus, use of the compositions of the present invention in combination with the firefly fluorescence of the da Vinci Surgical System provides an augmented view that enhances difficult to visualize tissue to reduce positive surgical margin rates, reduce adverse side effects, and potentially expedite surgery.

[0157] V. Examples

[0158] Example 1. Visualization of tumor tissue under irradiation

[0159] Four similar tumor tissues were treated with four different fusion compounds including different dyes and targeting ligands. The four fusion compounds included DyLight800 and PSMA-targeting ligand, IRDye800CW and PSMA-targeting ligand, ZW800 and PSMA-targeting ligand, and ICG and PSMA-targeting ligand. The treated tumor tissues of each of the four images were then exposed to narrow-band, near-IR excitation light to produce Figures 1A-1DThe narrow-band, near-IR excitation light includes wavelengths of light expected to correspond to the excitation maximum wavelengths of each of the fluorophores associated with each of the different compounds.

[0160] like Figure 2 As shown, the fusion compound including DyLight800 and PSMA-targeting ligand showed significantly more fluorescence. The significant difference in fluorescence may be due to at least one of the following reasons: (1) attaching ZW800, IRDye800CW, or ICG to PSMA-targeting ligand adversely affects the binding affinity of PSMA-targeting ligand to PSMA binding sites on tumor tissue; and (2) attaching ZW800, IRDye800CW or ICG to PSMA-targeting ligand adversely affects the fluorescence properties of the fluorescent moiety of ZW800, IRDye800CW or ICG. In contrast, conjugating DyLight800 to PSMA-targeting ligand does not have these adverse effects.

[0161] Example 2. Visualization of prostate tissue under irradiation

[0162] 20 mg of the compound of formula III in 10 ml of sterile water can be administered intravenously to the patient. The port of the laparoscope can then be placed and the da Vinci surgical system can be connected to the port. The endoscope of the system can then be guided to the patient's prostate, and laser excitation at about 800 nm can be used to excite the composition of formula III within the prostate. Small amounts of blue and green light can also be emitted to allow visualization of background anatomical structures. About 2-24 hours after administration, when the composition of formula III has been bound to PSMA, visualization of the prostate and prostate tumor tissue can be achieved.

Claims

1. Use of a composition for the preparation of a medicament for visualizing tissue expressing PSMA under irradiation with near-infrared radiation, wherein the composition is a pharmaceutical composition in a unit dosage form comprising a compound of formula III: The unit dosage form of the compound is in an amount between 0.01 and 8 mg / kg.

2. The use according to claim 1, wherein the unit dosage form of the compound is 0.01, 0.05, 0.10, 0.20, 0.30, 0.35, 0.40, 0.45, 0.50, 0.55, 0.60, 0.65, 0.70, 0.75, 0.80, 0.90, 1, 2, 4, 6 or 8 mg / kg.

3. The use according to claim 1, wherein the composition is in dry and single-dose form.

4. The use according to claim 1, wherein the composition is lyophilized in a sterile container.

5. The use of claim 1, wherein the composition is contained in a sterile container, and wherein the container has a machine detectable identifier that is readable by a medical device.

6. The use according to claim 1, wherein the composition is an oral dosage form and comprises a pharmaceutically acceptable excipient.

7. The use according to claim 1, wherein the composition is an injectable dosage form and comprises a pharmaceutically acceptable carrier.

8. The use according to claim 1, wherein the composition is in a dosage form comprising a pharmaceutically acceptable excipient for direct delivery to a surgical site.

9. The use according to claim 1, wherein the composition is sterile and non-toxic.

10. The use according to claim 1, wherein the tissue being visualized is cancerous tissue, wherein the cancerous tissue is prostate cancerous tissue.

11. The use according to claim 1, wherein the tissue being visualized is prostate tissue.

12. The use according to claim 1, wherein the tissue being visualized is prostate tumor tissue.

Images